Annals
of the
Missouri Botanical
Garden
Volume
: :1.Q14.-.
• V.
• * •
With Twenty-six Plates and One Figure
Published quarterly by The Rumford Press, Concord, N. H
for the Board of Trustees of the Missouri
Botanical Garden, St. Louis, Mo»
24085
Annals
of th
e
Missouri Botanical Garden
A Quarterly Journal containing Scientific Contributions
from the Missouri Botanical Garden and the Graduate Labora-
tory and Faculty of the Henry Shaw School of Botany of
Washington University in affiliation with the Missouri Botanical
Garden.
George T. Moore
Editorial Committee
Jacob R. Schramm
Benjamin M. Duggar
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♦ - ♦
ANNALS
OF THE
MISSOURI BOTANICAL GARDEN
TABLE OF CONTENTS
PAGE
Introduction i-ii
The effect of surface films
and dusts on the rate of
transpiration (with plate
1) B. M. Duggar and J. S. Cooley 1-22
Some pure culture methods
in the algae Jacob R. Schramm 23-45
The identification of the
most characteristic sali-
vary organism and its rela-
tion to the pollution of air
(with plate 2) August G. Nolte 47-80
The Polyporacese of Ohio. . L. 0. Overholts 81-155
A contribution to our knowl-
edge of the relation of cer-
tain species of grass-green
algae to elementary nitro-
gen (with one figure and
plate 3) Jacob II. Schramm 157-184
The Thelephoraceae of North
America. I. (with plates 4
and 5) Edward A. Burt 185-228
Indications regarding the
source of combined nitro-
gen for Ulva Lactuca .... G. L. Foster 229-235
The effect of certain condi-
tions upon the acidity of
tomato fruits B. M. Duggar and M. C. Merrill 237-240
A method for the differential
staining of fungous and
host cells R. E. Vaughan 241-242
TABLE OF CONTENTS
PAGE
Two trunk diseases of the
mesquite (with plates 6
^nd 7 Hermann von Schrenk
A trunk disease of the lilac
(with plates 8 and 9) Hermann von Schrenk
Descriptions of North Amer-
ican Senecionese (with
plates 10, 11, 12, 13, and
14) J. M. Greenman
A study of the physiological
relations of Sclerotinia cin-
erea (Bon.) Schroter J. S. Cooley
The ThelephoraceaB of North
America. IT. Craterellus
(with plates 15, 16, and 17) Edward A. Burt
The effects of surface films
on the rate of transpira-
tion: experiments with
potted potatoes (with plate
18) B. M. Duggar and J. S. Cooley
The Thelephoracese of North
America. III. Craterellus
borealis and Cyphella
(with plate 19) . Edward A. Burt
Some Oenotheras from Che-
shire and Lancashire (with
plates 20, 21, and 22) ... . R. R. Gates
A Texan species of Megap-
terium (with plate 23)... . R. R. Gates
Diagnoses of flowering plants,
chiefly from the southwest-
ern United States and
Mexico (with plates 24,
25 and 26) J. M. ^[reenman and C. H. Thompson
Enzyme action in Fucus ve-
siculosus L B. M. Duggar and A. R. Davis
General index to volume I
243-252
253-262
263-290
291-326
327-350
351-356
357-382
383-400
401-404
405-418
419-426
427-432
STAFF
OF THE MISSOURI BOTANICAL GARDEN
Director t
GEORGE T. MOORE
Benjamin M. Duggah,
Plant Physiologist.
Hermann von Schrenk,
Plant Pathologist.
Jesse M. Greenman,
Curator of the Herbarium.
Edward A. Burt,
Mycologist and Librarian.
Jacob R. Schramm,
Assistant to the Director.
Charles H. Thompson,
Assistant Botaniat.
Melvin C. Merrill,
Research Asisistant.
BOARD OF TRUSTEES
OF THE MISSOURI BOTANICAL GARDEN
President,
EDWARDS WHITAKER
Vice-President,
DAVID S. H. SMITH.
Edward C. Eliot.
George C. Hitchcock.
P. Chouteau Maffitt.
Leonard Matthews.
Saunders Norvell.
William H, H. Pettus,
Philip C. Scanlan.
John F. Sheplet.
EX-OFFICIO MEMBERS:
Edmund A. Engler,
President of the Academy of Science
of St. Louis.
David F. Houston,
Chanoellor of Washington University.
Henry W. Kiel,
Mayor of the City of St. Louis,
Herman Mauch,
President of the Board of Public Schools
of St, Louis.
Daniel S. Tuttle^
Bishop of the Diocese of Misaouri,
A- D. Cunningham, Secretary,
Annals
of the
Missouri Botanical Garden
Vol. I MARCH, 1914 No. 1
INTRODUCTION
In order to provide for the printing of scientific papers, which
formerly constituted a large part of the volume known as the
Annual Report, the Board of Trustees has authorized a new
journal, to be known as the Annals of the Missouri Botani-
cal Garden. The Annals will appear four times a year, in
March, May, September, and November, and contain only
scientific contributions from members of the staff of the Garden,
from the faculty and graduate students of the Henry Shaw
School of Botany of Washington University, and from vis-
iting botanists doing all or a part of their work at the Garden.
The increase in original contributions available for publication,
due to the additions to the staff and the greater number of
graduate students, makes it no longer possible to follow the
practice of the past and print papers from sources other than
the Garden.
The publication of a monthly bulletin by the Missouri
Botanical Garden, in which appear promptly the annual re-
ports of the officers of the Board, and of the Director, together
with popular accounts of the various activities of the Garden;
and the provision for the printing of scientific papers in the
Annals, has made it advisable to discontinue the Annual Report,
which was published each year from 1890 to 1912. The Twenty-
third Annual Report, therefore, marks the close of this series.
The Annals will be maintained upon a strict subscription
basis, using it in exchange only when its equivalent can be
obtained. Some of the institutions and societies, the publi-
cations of which have been received in exchange for the Annual
(i)
U INTRODUCTION
Report, issue nothing of value to a botanical library and appar-
ently are not interested in botany or related sciences. These
have been stricken from the exchange list. On the other hand,
the receipt of this number of the Annals is an indication that
the Missouri Botanical Garden desires to continue the old
exchange arrangement, the new journal being sent four times
a year in place of the old Annual Report. Additional exchanges
with publishers of journals dealing directly with botany are
desired. Upon request, the monthly Bulletin will be substi-
tuted for the Annals as an exchange with those desiring a more
F
popular and general account of the work and scope of the
Missouri Botanical Garden.
George T. Moore
Director.
1
THE EFFECT OF SURFACE FILMS AND DUSTS ON THE
RATE OF TRANSPIRATION
B. M. DUGGAR
Physiologist to the Missouri Botanical Garden
Professor of Plant Physiology in the Henry Shaw School of Botany of
Washington University
AND J. S. COOLEY
Rufus J. Lackland Fellow in the Henry Shaw School of Botany of
Washington University
The fungicides commonly employed are either in the form
of solutions (e. g., ammoniacal copper carbonate), suspensions
(lime wash and Bordeaux mixture), and powders (sulphur).
The use of spray mixtures or other fungicides has become
world wide, and many problems of physiological interest have
arisen respecting the effects of these substances on the plants
L
which they are designed to protect. Bordeaux mixture has
been under continuous observation for a period of about twenty
years, and has proved interesting in both its toxic and other
relations. The striking influence of this fungicide upon sound
plants has awakened widespread interest, and numerous
experiments have been made to determine the nature of the
effects. Bordeaux mixture consists essentially of suspension
films of copper hydroxid and certain other complex (mostly
hydrated), largely insoluble^ copper compounds; and when
properly sprayed upon plant surfaces from the best nozzles,
the particles are of extreme fineness, and there is realized an
almost perfect surface film. In spite of the greatest care in
preparation and application, it is injurious to certain plants,
such as the peach and the plum, and may not be used satis-
factorily in such cases for disease control. In recent years it
has been shown that the extent of the injury to the apple and
other plants may be considerable, and Bordeaux mixture is.
in such cases being supplanted. In this discussion, however,
we may omit any detailed consideration of the toxic effects-
of this mixture, a phase of the subject which has received much
consideration in this country from Bain (2), Crandall (6), Clark.
Ann. Mo. Bot. Gakd., Vol. 1, 1914
(1)
[Vol. 1
2 ANNALS OF THE MISSOURI BOTANICAL GARDEN
■
(4), Swingle (21), and others. Moreover, with the exception
of incidental references, we wish to deal at this time only with
its physiological action in prolonging the vitality of leaves
and plants.
During the first years of the use of this spray mixture it was
natural that any increased vitality of the sprayed plants would
be attributed merely to the action of the fungicide in restrain-
ing fungous or insect pests. Indeed, we find no authentic
suggestion of any other effect than that mentioned for eight
or ten years after the discovery of this fungicide. Since 1892
there have been frequent observations indicating beyond any
reasonable doubt that in the absence of all disease-producing
organisms there is often prolonged vitahty of the sprayed
plants as contrasted with the unsprayed. The increased
longevity is particularly noticeable in plants like the potato,
in which, under normal conditions, the foliage frequently dies
in advance of the first killing frost. Nevertheless, lengthened
life in leaves of deciduous trees, notably of the apple, has like-
wise been reported. It is not always possible to state definitely
to just what extent any apparent increased vitality is to be
attributed to the physiological action of the fungicide rather
than to the control of pests, and it must be said that the fre-
quency of the phenomenon and the reliability of the observers
alone preclude the possibility of constant errors in this matter.
In practical field experimentation the most significant differ-
ences in yield and vitality as a result of spraying with Bordeaux
mixture have been evident in the case of the potato, and with
this crop it is a matter of common observation both in Europe
and America. In recent years the consecutive reports on
potato spraying by F. C. Stewart (19) and his associates at
Geneva, New York, suggest in a decisive way the probable
magnitude of the Bordeaux influence when disease is a minor
factor. In general, observers are perhaps liberal in their
estimates of the gain from fungous suppression.
It will be pertinent to note a few observations and comments
from the reports of the work done at Geneva. In 1904 the
increase in yield from spraying potatoes five times was 233
bushels per acre. ''Spraying prolonged the life of the plants
25 days. Lateblight was the only trouble." In his experiments
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 3
of 1906 Stewart notes an increase in yield of 63 bushels per
acre due to spraying five times. He remarks: ''Late blight,
early blight, flea beetles and tip burn were all factors in this
experiment, but none of them caused much damage." More
striking were the results the following year when an increase in
yield of 73f bushels per acre was obtained from spraying five
times. In this case it is reported : "Late blight and rot were
wholly absent and early blight appeared only in traces. There
was some tip burn and a fight attack of flea beetles. Considering
the seemingly smafi amount of damage done by blight and
insects, it is remarkable that spraying should have increased
the yield so much." In 1909 the increase in yield in spraying
six times was 49f bushels per acre, and the comment upon this
result is as follows: "Early bfight, late blight and rot were all
absent. Some injury from flea beetles w^as noticeable through-
out the season. Ajfter September 1 there was considerable
tip burn. As late as September 24 the difference between
sprayed and unsprayed rows appeared sfight. The sprayed
row^s held most of their foliage until kiUed by frost on October
14."
The senior author of this report visited the experimental
plats which afforded these data in late September, 1911, prior
to the killing frosts of October 27, and the contrast between
the sprayed and unsprayed rows was pronounced; at the same
time there was very little evidence of any disease on the un-
sprayed plats. Regarding the condition of the plants Stewart
says: "There was no late bfight whatever, only a very fittle
early blight, and very little flea beetle injury. The unsprayed
rows were affected by no disease of any consequence except
tip burn, and even of that there was only a moderate amount.
As the plants were still partially afive twenty weeks after plant-
ing it is clear that they could not have been very much injured
by anything. Yet spraying increased the yield at the rate of
bushels
have here a striking exam
beneficial influence of Bordeaux in the absence of diseases
and insect enemies. "
Examining the comments of these and of other investigators
regarding increased vitality as a result of spraying with Bor-
deaux, we find that where the condition of the plant is well
[Vol. 1
4 ANNALS OF THE MISSOURI BOTANICAL GARDEN
defined at the close of the season, or at the time of the first
killing frost, the sprayed plants are almost invariably more
vigorous. Often, in the practical absence of any disease,
sprayed plants may remain healthy until killed by frost, while
unsprayed plants may have died from a few days to a few weeks
in advance of frost.
Following a recital of notable increases of yield in Connecticut
as a result of sprajdng potatoes with Bordeaux, Clinton (5) ex-
presses the conviction that an explanation must be found in the
conservation of water. His statement follows:
''The question naturally comes up, why did the sprayed
potatoes give this increased yield over the unsprayed if there"
was no particular injury caused by the late blight fungus?
Some little benefit was no doubt derived from the prevention
of the early blight, but this must have been scarcely appreciable
because this fungus was not at all conspicuous these years.
Again, some very small benefit may have been due to lessening
insect attack, since potatoes sprayed with both Bordeaux and
Paris green keep off the insects somewhat better than where
sprayed only with Paris green. This is especially true as regards
the potato flea beetle. But here again the gain was of a very
minor kind. . Ordinarily botanists have explained this increase
as due to some stimulative effect the Bordeaux mixture has on
the chlorophyll of the potato leaves in increasing starch pro-
duction. Personally, the writer believes that the results are
largely due to conservation of moisture in the leaves in dry seasons
by clogging up the stomata and water pores with the sediment of
the spray. The reasons for this belief are (1) that the potato
leaves, through their numerous stomata and terminal water
pores, lose water very easily, and are especially susceptible
to what is known as tip burn in dry seasons; (2) that the un-
sprayed vines uniformly suffered earlier and more severely
from tip burn than the sprayed, which were green for about
two weeks after the unsprayed were dead; (3) that in 1910,
which was a season like the preceding years, except with a
little injury from blight at the very end of the season, spraying
with 'Sulphocide' and commercial lime-sulphur, sprays with
comparatively little sediment, did not prolong the life of the
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 5
vines or give increased yield, while spraying with Bordeaux
mixture did."
Although this theoretical explanation did not come to our
attention until the experimental work reported in this paper
was complete, it was, in modified form, the only possible opinion
which we felt inchned to advocate, as a clue to the increased
longevity caused by Bordeaux, until the contrary evidence
yielded by our experiments.
Review of Literature
The experimental work undertaken in the past to determine
the nature of the Bordeaux influence (apart from direct injury)
has touched mainly upon (1) questions of increased photo-
synthesis due either to ''stimulation" of chloroplastid or
chlorophyll development, or to a direct influence upon light
quaUty; (2) changes in the respiratory rate, or surmised effects
upon metabolism; and (3) a modification of the normal rate of
transpiration. A few observations from the extensive liter-
ature with particular reference to its bearing on transpiration
may be cited.
Rumm (16) finds that in sprayed grapes the chlorophyll
content of the leaves increases and the fruit ripens earher with
a higher sugar percentage. He attributed these phenomena
to the higher ''assimilatory activity," and in turn relates this
to the following observation on transpiration: — that abscised,
sprayed twigs remain fresh longer than those unsprayed, from
which it is deduced that there is a falling off in transpiration
as a result of spraying. Through independent observations
made during the same year, Miiller-Thurgau (15) and Bayer
(3) subscribe to the view that lessened transpiration follows
spraying. Moreover, this confirmation of Rumm is obtained
by the former through an experiment which also proclaims
that the reduction in transpiration as a result of spraying
may be as much as forty per cent. Nevertheless, the report
referred to is extremely brief and does not indicate clearly the
condition of the plants during the period of observation, a
matter most important in the final interpretation of the data
afforded.
Frank and Kriiger (9, 10) reported some rather extensive
6 ANNALS OF- THE MISSOURI BOTANICAL GARDEN
[Vol. 1
quantitative experiments as a result of which they conclude,
contrary to Rumm, that transpiration is accelerated by spray-
ing. They state that sprayed leaves are in general more robust,
thicker and stiffer. They also report an increased yield in
pot experiments from spraying. All these indications, as well
as those of Leydheker (13) and others (1, 12) denote differences
of yield which are so slight as to be of no fundamental impor-
tance in the present consideration. Nevertheless, the trans-
piration data of Frank and Kruger, as already observed, were
obtained by satisfactory methods, and these are of greater
interest when taken in conjunction with those of Zucker (22)
who confirms their results entirely.
Schander (18) in an extensive paper reports a comparatively
small amount of experimental work on transpiration, but in
the cases given his results indicate a retardation of water loss
after spraying. His experiments with cobalt paper were incon-
sistent, and twigs of Taxus baccata and potted bean plants
were then employed, yielding the positive results noted. How-
ever, his work embraced very few plants, and the transpiration
differences observed are inconsiderable. He suggests that
lessened transpiration of sprayed plants is to be expected,
since the Bordeaux mixture must exert a shading influence as
a result of the exclusion from the leaf of certain injurious
rays. He attempts to verify this assumption of partial shad-
ing by a study of leaf temperatures, but the experiments in this
direction give no positive evidence for his theory. No adequate
mention is made of the conditions surrounding these experiments,
nor of the precautions observed.
Ewert's (8) experiments tend to substantiate the views of
Rumm and Schander; but, unfortunately, the results are
not satisfactory for accurate quantitative purposes, since
evaporation from the pots was merely checked and not pre-
vented,batting being employed to cover the soil surfaces. His
experiments are of particular interest, however, with respect
to his graph for comparative respiration in sprayed and un-
sprayed plants. In the sprayed plants, respiration was found
to be distinctly lower than in the unsprayed. It will be noted,
however, that this diminished respiration is scarcely in keeping
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 7
with the observation of Rumm and others regarding the higher
assimilatory activity in sprayed plants.
It is unnecessary in this report to review the considerable
literature which has accumulated bearing on the question of
increased starch formation as a result of the application of
Bordeaux mixture/ especially as it is proposed to discuss this
phase of the subject in a later paper.
METHODS
As indicated in the title, the experimental work here reported
is concerned merely with the transpiration of sprayed and un-
sprayed leaves or plants. Other effects of sprays and dusts
may be communicated in subsequent reports. In general,
the methods involved are modifications of customary practices.
The methods used were of two types, the experiments being
carried out either by means of (1) leaves in burette potometers
connected with side arm flasks, or (2) potted tomato plants.
Potometer Experiments. — After much preliminary experimen-
tation with a view to determining suitable leaves or twigs
for potometer work, leaves of the castor bean were selected.
Some of the preliminary experiments with other leaves are of
interest, however, and will be referred to subsequently. Castor
bean (Ricinus communis) leaves offer some special advantages,
especially (1) large surfaces, (2) resistance towards Bordeaux
mixture, and (3) prolonged vitality after abscision.
The burettes were connected with the side arm flasks, as
indicated in plate 1, and the flasks completely filled with water.
The petioles of the leaves were cemented into the mouths of
the flasks by means of "plastolina." If a ring of this plastic
substance is placed around the mouth of the flask when the glass
is dry and a ball of the same material, larger than the mouth
of the flask, is carefully attached around the petiole, then the
petiole and plastolina may be plunged into the mouth of the
flask and the two masses unite in a manner such as to give a
perfectly air-proof, water-tight connection. It has been found
desirable, for purposes of safety, to put on a second layer of
the plastolina as soon as it is evident that the first permits no
leakage. Even with these precautions, considerable diurnal
changes in temperature may cause leakage, and it is particu-
\
[Vol. 1
8 ANNALS OF THE MISSOURI BOTANICAL GARDEN
larly important that each experiment should be carefully ex-
amined prior to making all readings. The water columns
in the burettes were so gauged as to eliminate the possibility
of forcing water into the leaves. The burettes were employed
solely in order to get accurate readings of the water loss from
hour to hour without shifting or disturbing the plants by weigh-
ing; also rapidly to get data, should it seem necessary, under
changing conditions. All of these considerations proved very
important, as it was found that a slight shifting of the position
of the leaf affected materially the transpiration magnitudes.
For each leaf used it was necessary to get its rate of transpira-
tion in terms of some standard in order that the ratios might
be established between certain leaves prior to the addition
of the spray to some of them and the ratio between the same
leaves after the application. At one time it seemed possible
that the revolving table method of standardizing porous cups
might be applicable, but on further consideration it was believed
that the use of this method in the laboratory, and the subse-
quent disposition of the plants in the open, would lead to errors
of considerable magnitude. For our purpose it was not consid-
ered desirable to conduct the whole experiment on the revolving
table, but this method will be employed in connection with
our further studies. It was found very important to standardize
the leaf in a given position and then permit it to remain in
that position, as far as possible, throughout the experiment.
This method was necessary largely because of the fact that it
seemed wise to conduct the experiment in the open, during a
considerable interval, at least. Further reference to the ar-
rangement of the plants will be made in the discussion of the
experimental work.
Experiments with potted plants. — For the experiments with
potted plants tomatoes were used. The pots were dipped in
paraffin wax and the same sealing mixture was coated over
the surface of the soil. In all the experiments reported there
was no leakage in any case from improper sealing. Water
was added daily, or twice a day, to supply the loss by trans-
piration, the addition of water being made by means of a thistle
tube fixed in each pot. The bell of the thistle tube was covered
with paraffined paper during the entire interval. It was also
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 9
im
found necessary to insert in each pot a small bent tube in order
to provide for the changes in air pressure.
The pots were weighed at the beginning and at the close
of the experiment, but the condition of the plant and the amount
of water entering readily from the thistle tube were found
adequate to indicate the daily water requirements. To the
total provisional transpiration quantities obtained from a
summation of the quantities daily added, the differences in
weight between the beginning and the close of the experiment
were added or subtracted as required. From five to ten plants
were employed with each kind of spray or dust used, and the
plants of each lot were so distributed in the greenhouse that
an equal number — so far as possible — from every group was
subject to exactly the same influences. Moreover, positions
in the greenhouse were shifted several times during the obser-
vation intervals of from ten days to two weeks. As a result
of a large amount of experimental work in the greenhouse
it has become apparent that the points just referred to are
)ortant. Plants situated nearer the edges of the benches,
or those which receive drafts from opening doors or from
convection currents, show considerable differences in trans-
piration rates, and this should be obviated.
The leaves in the potometer work and the potted plants
were sprayed or dusted liberally, and in the case of the sprays,
in particular, care was taken to cover completely with a fine
spray of the material both surfaces of the leaves. The dust
applications were made in the late afternoon when the leaf
surfaces were less dry, and after dusting the upper surfaces
of the leaves the plants were inverted and the lower surfaces
equally well treated. The dusts were prepared by grinding
to an impalpable powder in a mortar.
The Bordeaux mixture employed was made by the 4-6-50
formula, the weights of ingredients for making small quaritities
being approximately as follows:
CuSOi 9 . 6 grams
CaO 14 . 4 grams
Water 1000 cc.
The weak Bordeaux was one-half the strength of the above.
The Ca(0H)2 was prepared by slaking gradually 60 grams of
10 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol, 1
CaO in 1 liter of water; and the mixture designated A1(0H)3
was prepared by mixing two solutions each of 900 cc., the one
containing 26 grams of AICI3 and the other 30 grams of CaO
(slaked as for the Bordeaux mixture). The clay suspension
consisted of 90 grams of fine air-dried clay in 1 liter of water.
The lime-sulphur employed was the usual 1-25 strength.
EXPERIMENTAL DATA AND DISCUSSION
It will be observed from the brief review of earher work
that the evidence regarding the effect of Bordeaux mixture
on the transpiration rate is inconsistent. A majority of the
observers adopt the view that the effect of this surface film
is to reduce the transpiration. On a priori grounds this view
would seem to be logical^ since it would indicate a water conser-
vation to which, in dry seasons at least, the plant might respond
with increased vitality and yield. Nevertheless, it was believed
that the experimental evidence at hand was of insufficient scope
to establish this view of it. Contrary to expectations, all of
our more important experimental evidence and observations
are antagonistic to the a priori assumption as applied to the
effects of Bordeaux mixture.
Potometer experiments. — In attempting to secure leaves sat-
isfactory for the work, some incidental observations were made
which are of interest. The work was begun during the winter,
so that greenhouse-grown plants alone were available.- Fur-
thermore, in this work with potometers, Bordeaux mixture
alone has been used by us. Testing leaves of squash {Cucur-
bita sp.), Pelargonium zonata, and Phytolacca, also shoots of
potato and Irescene, as to their behavior under the conditions
required, it was found that of comparable leaves, sprayed and
unsprayed, invariably the sprayed leaves were the first to wilt.
This might be attributed either to an injurious action of the
spray or to a greater water consumption. That the last men-
tioned is the more probable explanation finds confirmation
through a special observation on the potato. Owing appar-
ently to some stoppage of the vascular system, abscised potato
shoots are unsuitable for potometer w^ork, wilting in a com-
paratively short time even when cut under water; and sprayed
potato shoots wilt more quickly than unsprayed, thus pointing
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 11
to a more rapid water elimination after spraying. Potted
potato plants from which the shoots were cut withstood the
fungicide satisfactorily.
Leaves of the large elephant's ear {Caladium sp.) proved
unsatisfactory on account of the excessive "bloom," which
interfered with the proper application of the spray. Canna
leaves were similarly unfavorable, and leaves of the calla lily
wilt soon after abscision.
It has been stated above that the leaves of the castor bean
proved most satisfactory in the potometer work. The experi-
ments with these leaves were carried out in the open, except
as otherwise noted, during the early fall. The plants were
arranged for standardization and for subsequent observation
at distances of about ten feet apart on an exposed lawn uni-
formly sodded. No readings were made until the leaves had
become adjusted to the conditions. Observations were made at
frequent intervals when the water loss was rapid, in order to
maintain the water column at a fairly uniform level, so that
many of the data given in the tables which follow represent
summations of several successive readings. Three series of
potometer experiments were made, each series embracing six
leaves, but in one series, accidents to some of the leaves, and
the necessity of substituting new ones after the experiment
began, resulted in such a shortening of the standardization
intervals that it was thought necessary to discard the results,
although they were in the same direction as the others obtained.
The data are presented in full in the tables and all available
data are used in computing the relations given. The relations
may be more conveniently expressed if we first divide the
leaves into classes, designated by letters, as follows :
A — , three leaves (i. e., the transpiration quantities of these)
in the standardization interval before spraying.
A + , the same three leaves as in A — , but for any interval
after spraying.
B, three control leaves (unsprayed throughout) during the
standardization interval.
B\ the control leaves as in B, after standardization.
A — A +
The ratio -~^ = Q is to be compared with the ratio "gT'^Q'-
[Vol. 1
12
ANNALS OF THE MISSOURI BOTANICAL GARDEN
If Q' is greater than Q, then the spraying facilitates transpira-
tion; if less, then the converse is true. If no accidents occurred
during the experiments,
A
B
would, of course, be a constant
quantity, each term referring properly to the summed trans-
piration quantities for three leaves during the standardization
interval. Accidents are unavoidable, however, during the
subsequent observations, and whenever these occur it is neces-
sary to compute a new value of Q' for any particular "run"
in which the accident occurs. The only consideration then is
to have the same leaves (i. e., their summed transpiration
quantities) in the ratios before and after standardization,
for example, it is necessary to use a ratio, Q', of Nos. 1 and 3
to Nos. 2, 4, and 6 after spraying, then the new value of Q
(in the standardization interval) for comparison must also
be computed with Nos. 1 and 3 against Nos. 2, 4, and 6.
TABLE I
EFFECT OF FILMS OF BORDEAUX MIXTURE ON TRANSPIRATION OF STANDARDIZED
CASTOR BEAN LEAVES; DATA FOR DAY PERIODS
No. of leaf
1
2
Transp. 12:30-2 :30 P.M.,
Ist day before spraying
A-
10.8
3
Transp. 3:12-5:00 P. M.,
Ist day after spraying
.4 +
7.6
B
17.3
B'
20.4
A-
28.5
A +
23.2
4
B
45.9
5
B
33.8
B'
26.2
6
Ratio
A-
33.6
A
B'
26.1
B
72.9
97.0
4 +
39.5
Relation, sprayed to un-
spraycdj Ist day
Transp. 8:12-9:48 A.M.,
2nd day after spraying
72.9
Rate ohanged from Q7~n ^ ~ ■ ^^) ^^
A+ 70.3
B' ~72.7
70.3
72.7
(=.97)
Relation, sprayed to un-
eprayed, 2nd day, a.
A +
10.2
B'
37.7
63.1
B'
32.3
B'
29.7
A +
67.1
A+ 140.4
B'
99.7
72. 9
Rate changed from ^^^.tt^ (
Transp. 11 :16-11 :53 A. M.,
2nd day after spraying
Relation, sprayed to un-
sprayed, 2nd day, h.
97.0
140.4
99.7
.75) to
( = 1.41)
A +
4.9
B'
5.9
A4-
B'
31.5 13.8
B'
7.5
A +
25.3
^+ 61.7
H' "27.2
Rate changed from „_ ' (
.75) to
61.7
27.2
( = 2.3)
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION
13
TABLE II
EFFECT OF BORDEAUX MIXTURE ON TRANSPIRATION OF STANDARDIZED CASTOR
BEAN LEAVES; DATA FOR DAY AND NIGHT PERIODS
No. of leaf.
1
Tranap. 4:04-5:25 P. M.. 1st
day before spr.
A
7.5
Transp. 8:21-11:17 A. M.,
2nd day before spr.
Total transp. before spr.
20.2
2
B
7.6
3
4
5
10.9
B
11.7
A-
7.6
B
30.7
A-
27.7
Transp. 12:30-4:50 P. M.J A +
Ist day after spr, 1 36 . 7
B
38.3
A-
32.4
43.3
B
40.9
A-
23.3
B
52.6
A-
29.9
B'
42.6
A +
62.9
B'
50.2
A +
41.0
6
B
9.4
B
17.9
Ratio
B
27.3
B'
30.2
Relation, sprayed to un-
sprayed, 1st day
Transp. 8:56 A. M., to 4:44
P. M., 2nd day aft. spr.
Rates changed from ^^^'^ ( == . 86) to -tt^ { = 1 . 14)
125.2
123
Relation, sprayed to un
sprayed, 2nd day
Transp. 10:27 A. M., to 3:40
P. M., 3rd day aft. spr.
Relation, sprayed to un-
sprayed, 3rd day
Transp. 9:58 A. M.. to 4:42
P.M., 4th day aft. spr.
* Relation, sprayed to un-
Bprayed, 4th day
Total transp. after spraying
A +
20.7
B
A +
28.9
18.2
A +
17.0
B'
12.1
Rates changed from ^^ (=1.28) to ^ (=2.2)
79.9
30.3
A +
8.4
B
A +
11.2
B'
4.4
A +
7.8
B'
3.5
A
101.9
B 118.2
A+ 140.6
B'
123
A+^66.6
B' 30.3
A+ 27.4
B'^ 7.9
101 Q 27 4
Rates changed from ^^^ (=1.28) to 4~ (-3.46)
79.9
7.9
A +
B
A-\-
22.1
B'
14.1
A +
15.3
B'
7.9
A+ 37.4
B'~22.0
74 2 37 4
Rates changed from =rr-^ ( = . 93) to 7~^ ( = 1.7)
79.9
22.0
Relation, sprayed to un-
sprayed, totals
■
A +
125.1
B'
86.9
A +
81.1
53.7
A+ 206.2
B' "140.6
Rates of totals changed from ;^;^ (=. 93) to 77777-7; (—1.47)
79.9
140.6
Transp. 5:30 P. M., to 8:21 A
A. M., 1st night bef. spr.
Transp. 4:50 P.M., to 8:40
A. M., Ist night aft. spr.
6.9
A +
6.9
B
6.3
B'
7.8
A-
9.0
B
11.2
A-
6.4
B
A
22.3
B 17.6
A +
2.7
B'
7.5
A +
6.2
3.5
Relation, sprayed to un-
sprayed (night)
22 3 15 8
Rate changed from ,V^ (=1.27) to tt^ ( = 1.03)
17.5
15.3
Transp. 3:45 P. M., to 9:30
A. M., 2nd night aft. spr.
A +
20.0
B'
A +
15.1
B'
4.9
A +
5.8
B'
4.2
Relation, sprayed to
sprayed (night)
un-
22 3 40 9
Rate changed from ^rr^ (=2.0) to ^t-tt (=8.34)
11.2
4.9
A+ 15.8
B' "'15.3
A+ 40.9
B' "4.9
For this *'run" the plants were transferred to a room in the building.
[Vol. 1
14 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Summarizing the data for the rates in table ii, day intervals,
we find that Q.Q', in the successive periods, as .86:1.14, as
1.28:2.2, as 1.28:3.46, and as .93:1.7. If we make the ratio
before spraying equal in each case, to 1.0, then the value for
the periods after spraying in the successive day intervals are
respectively 1.33, 1.72, and 1.83. These differences in rate
are so marked and consistent as to outweigh all considerations
of individual differences, as disclosed by a detailed study of
the figures in table ii. It will also be noted that the less
extensive data from table i are confirmatory; thus Q:Q',
in the successive intervals, as .75:. 97, as. 75: 1.41, and as .75:2.3.
On the basis of 1.0 for the ratio before spraving, we have for
the Deriods after
29. 1.88, and 3.07
From the records of the potometer experiments it is obvious
that only one conclusion may be drawn, namely, that the rate
of transpiration is materially increased after spraying.
Some points relative to environmental conditions, however,
require special mention, and certain suggestive results must
be left for further experimental study. Attention has been
drawn to the fact that, in general, the potometer experiments
were conducted in the open, during early October. During
the last days of the work, cooler weather and danger from
rain made it desirable to transfer the potometers to a room in
the building, and the data for the third and fourth days after
spraying, table ii, were secured under these new conditions.
In this room the shades were drawn and every precaution
taken to secure uniformity. It will be noted that while the
order of results is in the same direction as for the lawn exposure,
the ratio is even higher than the average. No "shading action"
of the Bordeaux, as postulated by Schander (18), could be
considered a factor of importance in this case.
The results in the laboratory suggest, further, that the ratio
of sprayed to unsprayed will vary considerably with the con-
ditions. Before removing the potometers to the laboratory,
ic r>
the night temperatures were so low that two night "runs
(including the interval from about 6 p. m. to 8 a. m.) were
necessarily excluded on account of leakage. Other night "runs,"
as shown in the tables, indicate the probability that under
r
certain conditions unfavorable for evaporation^ the surface
19141
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 15
film may actually effect a diminution in the rate of transpira-
tion, although the transpiration data do not suffice to warrant
more, at present, than an impression. In fact, the night
"runs" should be considered apart from those of the day, for
the latter are much more satisfactory.
Experiments with potted plants. — The experiments with potted
tomato plants were divided into two series which were consec-
utive in time, and different only with respect to the substances
applied to the leaves. As far as has been ascertained, this is
the first time that tomato plants have been used in such work,
but in our experience they are more satisfactory than potatoes.
In the first series (table iii) 30 plants were used, in lots of
10 each, for the applications of (1) strong Bordeaux mixture,
(2) weak Bordeaux, and (3) controls. In the second series
(table IV, v) 80 plants were used in 8 lots, and the substances
employed as sprays or dusts are noted in the tables. In the
second series it is to be noted that there are 3 substances of
the nature of films (Ca(0H)2, A1(0H)3, and lime-sulphur), 1
true suspension (clay), and 3 powders (charcoal, CaCOa, and
powdered A1(0H)3).
The methods of procedure involved in these experiments
have already been outlined. It is necessary to add, however,
that the plants used were about 12 inches high and as uniform
in size as could be obtained. It was not possible satisfactorily
to standardize plants for an experiment extending over several
weeks : and it was necessary to rely in part upon numbers,
and in part upon a rigidly accurate method of selecting the
individual in each lot to eliminate any errors. The method
of selection consisted in getting together 8 plants so similar
in size and vigor that no choice could be made between them,
then distributing these at random to the 8 lots, this being
continued until each lot embraced 10 plants.
In each case the experiments extend over 2 periods. At
the close of the first period the plants were shifted in position
and a second application of the spray mixture or dust was
given. With the conclusion of the experiment the green weights
of all plants were taken, thus enabling us to determine, in
addition to the total transpiration quantities, the amount of
transpiration per gram of green substance.
2
[Vol. 1
16
ANNALS OF THE MISSOURI BOTANICAL GARDEN
TABLE III
THE EFFECTS OF BORDEAUX MIXTURE ON THE RATE OF TRANSPIRATION; DATA
IN GRAMS FOR 30 POTTED TOMATO PLANTS
Transpiration quantities
571
628
356
559
549
554
.
575
442
368
. 574
603
518
515
720
385
740
453
420
514
499
437
570
534
417
J
495
702
439
546
• ■ « «
564
Total
5659
5130
4458
Ave. per plant
F
565.9
570
445.8
1
1
1st period Oct. 18 to Nov. 4
Covering
Strong Bord.
Weak Bord .
Check
Plants nos.
1-10 11-20
21-30
Transpiration quantities
702
681
390
\
684
651
555
665
540
375
w
630
585
525
625
857
395
710
440
465
640
685
415
445
648
365
645
645
490
560
• • • «
545
1
1
Total
6306 6622
4520
Ave, per plant
630.6
625
452.0
'
Second perio
d Nov. 5-15
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION
17
TABLE III (Cordinued)
THE EFFECTS OF BORDEAUX MIXTURE ON THE RATE OF TRANSPIRATION; DATA
IN GRAMS FOR 30 POTTED TOAIATO PLANTS
Green wts. of plants used.
Plants nos.
1-10
21-30
Green weights in grams
55
80
49
58
^
58
63
51
41
46
■
39
51
56
50
60
39
67
45
53
46
40
48
46
53
40
i
' r
39
74
48
49
* *
55
Total
500
502
497
Ave. per plant
50.0
55.8
49.7
TABLE IV
THE EFFECT OF VARIOUS SPRAYS AND DUSTS ON THE RATE OF TRANSPIRATION;
DATA IN GRAMS FOR SO POTTED TOMATO PLANTS. 1ST PERIOD, OCT. 25
TO NOV. 8
1
1
r 1
^H ^P^k. ^^^P-^^^ ■-
.^IBlJ ^
Lime-
Covering
Ca(OH),
A1(0H),
Clay
Al(OH),
pwd.
Char-
coal
CaCO,
sulfur
1-25
Check
Plants nos.
30-39
40-49
50-59
60-69
70-79
1
80-89
1
1 " " "
100-109 90-99
Transpiration
431
394
345
416
378
436
508
352
quantities
437
370
333
370
460
353
414
323
435
386
430
393
374
315
474
461
411
383
383
383
460
510
354
443.
358
329
347
645
273
375
526
490
372
377
520
449
467
346
421
309-
-
314
398
437
365
320
471
352
330-
1
416
410
560
531
359
361
346
323-.
375
517
362
408
386
456
285
343 -
1
1
485
460
452
412
331
364
402
317:
Totals
4034
4024
4169
4372
3808
3987
4082
3691
Ave. per plant
403.4
402.4
416.9
437.2
380.8
398.7
408.2
369.1
[Vol. 1
18
ANNALS OF THE MISSOURI BOTANICAL GARDEN
TABLE V
THE EFFECTS OF VARIOUS SPRAYS AND DUSTS ON THE RATE OF TRANSPIRATION;
DATA IN GRAMS FOR 80 POTTED TOMATO PLANTS. 2ND PERIOD, OCT. 25
TO NOV. 8.
^
M
A ^K d ^^^ ^^^^^^^m ^
^ii^^ ^
4
Lime-
4
Covering
Ca (OH) ,
A1(0H),
Clay
A1(0H).
pwd.
Char-
coal
CaCO 1
sulfur
1-25
Check
Plants nos.
30-39
40^9
50-59
60-69
1
70-79
80-89
100-109 90-99
Transpiration
494
1
812
624
1
665
573
1
560
846
690
quantities
463
587
682
601
610
543
641
569
592
653
1
693
609
530
553
667
564
539
587
615
654
744
700
622
701
604
654
594 ■
754
556
512
680
764
457
614
579
606
617
552
616
546
544
665
694
476
569
706
604
496
647
605
753
704
664
685
478
558
597
806
579
641
596
620
601
514
582
810
690
711
562
617
428
601
Totals
5509
6793
6303
6421
6021
5948
6183
5903
Ave.
550.9
679.3
630.3
642.1
602.1
594.8
618.3
590.3
1
1
*Transp.
pergm.
11.8
12.2
12.3
11.4
13.1
12.0
12.8
12.1
Green weights
in grams
Totals
Ave. per plant
Green wts. of plants 30-109 at close of 2nd period.
39
39
49
42
60
43
41
50
60
43
466
46.6
71
43
58
63
56
56
53
55
58
64
557
55.7
46
47
61
49
54
58
52
50
48
59
514
51.4
■(
I
58
52
45
79
77
47
47
54
47
65
561
56.1
49
43
48
35
35
59
46
48
52
43
458
45.8
50
47
43
63
53
51
57
47
43
51
495
49.5
66
68
51
47
42
38
53
67
67
63
40
52
36
49
39
485
48.5
66
35
46
46
51
487
48.7
Computed on the basis of green weights at the close of second period.
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 19
An examination of the data in the several tables involved
in the pot experiments serve to indicate that while there is a
certain amount of individual variation in the transpiration
quantities of the various plants in any group, general conclu-
sions seem to be warranted. The individual variations in
transpiration in the Bordeaux series are in closer accord with
the variations in green weight of the plants used than are those
in the other series. Taking all factors into consideration, a
film of Bordeaux mixture is found to facilitate transpiration.
Other films and dusts employed do not seem to affect the rate
of transpiration to the same extent.
In a consideration of the results in detail it is to be noted
that the Bordeaux series (table iii) is not strictly comparable
with the other (tables iv, v), since they were not conducted
simultaneously. If the transpiration in grams per gram of
green weight for the control (Bordeaux series) is represented
by 100, then the rate for weak Bordeaux on this basis is 113.2,
and the rate for strong Bordeaux is 125.43. The differences
are in the same direction, but not so great as those obtained
with the potometer experiments. The use of both weak and
strong Bordeaux mixture materially strengthens the conclu-
sions to be deduced.
The series which gives the results with other sprays and
dusts is not so easily interpreted. The transpiration quantities
vary sHghtly on either side of the control, and no covering
gives a negative difference (contrasted with the control) greater
than six per cent (this is the case of A1(0H)3), or a positive
difference greater than about eight per cent (charcoal).
These slight average differences may be no more than would
be explained by the possible experimental error; but it is of
interest to perceive that, with the exception of clay, those
surface applications which give lower values than the control
are those which might diminish the absorption of heat in direct
sunshine. The results might then be the resultant of two
factors, (1) the direct effect of the surface film or dust on the
rate of water loss, and (2) the indirect effect exerted through
a modification of the temperature of the leaf.
Accepting as a general conclusion an acceleration of trans-
piration (specifically in the castor bean and in the tomato)
20 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
folio
■esult of an application of a film of Bordeaux mixture, the
ving questions arise: (1) What is the physical or chemical
basis of the increased evaporation from plant surfaces covered
with Bordeaux mixture? (2) Is the increased evaporation in
any way related to the increased vitahty or longevity of sprayed
leaves? Neither of these questions may be answered intelli-
gently at present. With respect to the first, we have arranged
experiments to determine the effects of Bordeaux on the passage
of water vapor through non-living membranes; but the results
are thus far conflicting, due possibly to the fact that we have
not yet used membranes which are satisfactory analogues of
leaves. Experiments in this direction will be reported later.
No relation of transpiration to increased longevity can be
foretold, although it seems possible that the highest efficiency
equilibrium relation of longevity may involve, in certain plants,
a relatively high transpiration rate as either a direct or an
indirect factor. No answer to the question will be satisfactory
until a further study of other effects (''stimulation," increased
''assimilatory activity," etc.) of Bordeaux mixture shall have
been made.
Bibliography
1. Aderhold, R. Der heutige Stand unserer Kenntnisse uber die Wirkung und
Verwertung der Bordeauxbriihe ala Pflanzenschutz-mittel. Jahresb. d. Verein-
igung
1904.
2. Bain, S. M. The action of copper on leaves. Tenn. Agr. Exp. Sta. Bui. 15:
1-108. 1902.
3. Bayer, L. Beitrag zur pflanzenphysiologischen Bedeutung des Kupfera in der
Bordeauxbrtihe. Inaug-Dissert., Konigsberg, 1902 [cf. Schander (18)].
4. Clark, J. F. On the toxic properties of some copper compounds with reference
to Bordeaux mi>ture. Bot. Gaz. 33: 26-48. 1902.
5. Clinton, G. P. Spraying potatoes in dry seasons. Conn. Agr. Exp. Sta. Report
1909-10: 729-752.
6. Crandall, C. S. Bordeaux mixture. 111. Agr. Exp. Sta. Bui. 135 : 201-296.
1909.
7. Duggar, B. M. Peach leaf curl and notes on the shot-hole effect of peaches and
plums.
8. Ewert, R.
164: 371-388. 1899.
Kupf erkalkbr iihen
auf den Stofifwechsel der Pflanze. Landw, Jahrb. 34 : 233-31 1 . pi. 2-4. 1905.
9. Frank, A. B., and Kriiger, Fr. Ueber den direkten Einflusa der Kupfervitriol-
10.
kalkbriihe auf die Kartoffelpfianze.
1-46
Kartoffelpfianze hervorbringt.
Behandlung
8-11. 1894.
11. Hawkins, L. A. Some
Bur. PI. Ind. U. S. Dept. Agr. Bui. 265: 1-29. 1912.
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION ^i
12. Kirchner, 0. Ueber die Beeinflussung der Aseimilationstatigkeit von Kartoffel-
pflanzen durch Bespritzung mit Kupfervitriolkalkbriihe. Zeitschr. f . Pflanzenkr.
i8: 65-81. 1908.
13. Leydheker, A. Die Bekampfung der Kartoffelkrankheit durch die Verwendung
von Kupfervitriol. Oesterr. landw. Wochenbl. 1893 : 163. [Reviewed in
Zeitschr. f . Pflanzenkr. 4 : 33. 1894.]
Lutman, B. F. The covering power of the precipitation membr
mixture. Phytopathology 2: 32-41. 1912.
MuUer-Thurgau, H. Jahresb. der schweizerischen Versuchssta
f . Obst-. Wein- und Gartenbau in Wadensweil 1892-93 : 68-59
und
16. Rumm, C. Ueber die Wirkung der Kupferpraparate bei Bekampfung der Boge-
nannten Blattfallkrankheit der Weinrebe. Ber. d. deut. bot. Ges. 11: 79-93.
1893 .
17. , Zur Kenntnifls der Giftwirkung der Bordeauxbriihe. Inaug-Dissert.
1-76. 1 pi. 1895.
18. Schander, R. Ueber die physiologische Wirkung der Kupfervitriolkalkbruhe.
Landw. Jahrb. 33 : 517-584. 1904.
19. Stewart. F. C.. French, G. T., and Sirrine, F. A. Potato spraymg expenments
m 1910.
20
58: 115-151. 1910.
periments, 1902-1911. N. Y.
(Geneva) Agr. Exp. Sta. Bui. 349:99-139. 1912.
21. Swingle, W. T. Bordeaux mixture: its chemistry, physical properties, an
effects on fungi and algae. Div. Veg. Physiol, and Path. U. S. Dept. Ag
9:1-37. 1896.
22. Zucker, A. Beitrag zur direkten Beeinflussung der Pflanzen durch die 1
vitriol-Kalkbriihe. Inaug-Dissert. Stuttgart, 1896 [cf. Schander (18)1.
Graduate Laboratory, Missouri Botanical Garden.
22
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1, 1914]
Explanation op Plate
PLATE I
etcr used in the transpiration experiments showing burette connected
arm flask, and abscised leAf nf Rrr^'nitQ i^pmAn+^rl \r^irx fu« w,^,,+u
plastolina.
y ^
'- J
K k
Ann. Mo. Bot. Gard., Vol. 1, 1914
Plate I
\
DIGGAR AND COOLEY— TRAXSIMRATIOX
SOME PURE CULTURE METHODS IN THE ALG^
JACOB R. SCHRAMM
Assistant to the Director of the Missouri Botanical Garden
Instructor in the Henry Shaw School of Botany of Washington University
Introduction
Too much confidence has frequently been placed by algolo-
gists in their ability to recognize a given species of alga among
varying numbers of other species, and in the various forms
which it may assume — a fact which has led to much confusion
and error, especially among members of the Protococcales. While
it is now definitely known that in a number of algse a single
species may present markedly dissimilar appearances, either
as a result of varying environmental conditions, or because of
the presence in the fife history of several unlike stages, it is cer-
tain that much of the so-called polymorphism , or pleomorphism,
of algse finds its explanation in inadequate methods of study.
It is becoming recognized that for life history studies in the
algse it is necessary to employ cultures free from other species
of algse. Even in cases where this is not, on first thought,
necessary, as in the large, filamentous forms, it should be ob-
served, for the possibihty of introducing spores or sporelings
of closely allied species is by no means excluded in all cases.
Gratifying progress has already been made by some algologists,
working especially with members of the Volvocales and Pro-
tococcales, and it seems reasonably certain that the originally
chaotic condition existing in the latter will be ultimately reduced
J
to complete order by a careful observance of the necessity of
working with pure cultures, or at least cultures containing but
a single species of alga. In life-history studies where physio-
logical differences between species are to be investigated, it
is especially desirable and indeed necessary to employ pure
cultures.
Certain species of algae, especially representatives of the
ChlorophycecE, have been much used in physiological investi-
gations — chiefly those concerning themselves with various
Ann. Mo. Bot. Gaed., Vol. 1, 1914
(23)
(Vol. 1
24 ANNALS OF THE MISSOURI BOTANICAL GARDEN
phases of nutrition. With the development of a clearer under-
standing of the activities and life processes of the various
micro-organisms, the necessity of working with rigorously
pure cultures has become more and more evident. It is now
generally appreciated that, in most cases, vaUd conclusions as to
the physiology of a particular organism cannot be drawn with
certainty where one or more foreign organisms have been present
in the cultures. There can be no doubt that the frequent con-
tamination of cultures of algse with bacteria, and even with
fungi, has, in many cases, detracted markedly from the value
of painstaking and otherwise careful physiological investigations.
The readiness, however, with which many algse lend themselves
experimental
of their small size and
ease of handhng and culture — will always make them favored
objects of study; and it appears desirable at this time to bring
together some of the experiences of the author in the prepara-
tion of pure cultures of algse, with the hope that suggestions
may be gained from them by those who desire to obtain such
cultures for one purpose or another.
An unfortunate use of the term "pure culture" has come
into more or less general use and has frequently led to confusion
and ambiguity. As used by many authors, it means simply
a culture of a single species of alga not necessarily free from
bacteria and fungi. Where the presence of other organisms
is not specifically mentioned, it is clear that the above usage
of the term may lead to serious misunderstandings. Indeed,
it remains for the reader, in many instances, to decide for
himself — from the technique employed — whether a culture of
an alga free from all other organisms or only from other species
of algse is meant. It is to be hoped, therefore, that the term
pure culture shall come to have the same clearly defined mean-
ing when used in connection with the algse that it has long had
in the fungi and bacteria. In the following report the term
is used to signify a culture of a single species of alga free from
other organisms
r
Historical
■
Although incidental references to pure culture technique in the
algse are frequently found in the literature, relatively few con-
tributions have appeared which deal extensively with the
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^E 25
subject, or which outline in detail the methods employed.
Beyerinck, in 1890 (4, 6), appears to have been the first to
succeed in isolating species of algaj in pure culture. Ditch
water, boiled with ten per cent gelatin^ and cooled, was mixed
with a drop of water rich in protococcoid algse, poured into
dishes, and allowed to cool. Numerous minute algal colonies
appeared in course of time, and the number of bacterial colonies
developing was so small that successful transfers of Scenedesmus
acutus Meyen and Chlorella vulgaris Bey., were made, both
organisms being subsequently cultured on a variety of media.
In addition, the gonidia of Physcia parietina were obtained
pure. Small pieces of the Uchen thallus, carefully washed,
were placed on soUd gelatin plates. Those which showed
themselves to be free from foreign organisms were transferred
to gelatin plates containing malt-extract, the fragment being
first torn to bits with needles and then dragged over the sterile
surface. In a few days, small colonies of the algal symbiont
appeared from which successful transfers were made. In a
later paper (5), Beyerinck adds Stichococcus major and a second
species of Chlorella to the list of algse previously cultured in
a state of purity, the technique, in general, being the same.
Miquel (16) was the first to isolate a diatom in pure culture.
Subsequently, Richter (20, 21) isolated Nitzschia Palea (Kiitz.)
W. Sm., and Navicula minuscula Grun., by the use of synthetic
agar plates. Attention is called by this author to the impor-
tance of using agar which has previously been washed to free
it from soluble impurities. A mixture of diatoms and other
algae was placed on the surface of washed agar plates, and from
the impure diatom colonies which developed transfers were
made to other plates until at length pure cultures were obtained.
In his isolations of
(18) also obtained Polyioma uvella. While his method seems
unnecessarily complex, it is of interest here. Sterile capillary
tubes were filled in part with a column of sterile water, and
subsequently a column containing the organisms was added
below, care being taken not to separate the two by air. Both
ends of the tube were then sealed. After sufficient time had
elapsed for the movement of the motile Polytoma cells from the
lower column into the upper sterile one, the tube was broken in
26 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
the region of the upper column. The lower portion was dis-
carded, and the upper one was sealed, subsequently transferred
to a sterile medium, and broken to permit the organisms, free
from contaminations, to enter the medium and begin their
development.
By the gelatin plate method, Kriiger (13) prepared pure
cultures of two new organisms— C/iZoreZZa protothecoides and
C hlor othecium saccharophilum — obtained from the exudation
of Populus alba. Tischutkin (23) lists representatives from
about eighteen genera of algsB — including diatoms, green, and
blue-green forms — as having been obtained in pure culture
by the agar plate method. After three or four successive dilu-
tions in Hquid one per cent agar, the organisms were plated in
Petri dishes. The filamentous forms he washed in sterile water,
cut into short segments, and transferred to the liquid medium.
The methods given by Ward (24) include plating in agar and
silicic acid jelly, though as a whole the methods are applicable for
the separation of algal species rather than for their isolation in
pure culture. This is especially true of the plaster of Paris, and
precipitated calcium carbonate methods. Gonidia from Xan-
^
thoria parietina, and Gasparinia murorum (Hoffm.) Tornab., to-
gether with Pleurococcus vulgaris and Scenedesmus caudatus were
obtained in pure culture byArtari (1). Chodat and Goldflus (8),
by the use of pieces of sterilized unglazed porcelain in contact
with a mineral nutrient solution, claim to have isolated a species
of Nostoc in pure culture. The procedure was a simple one,
consisting in repeated transfers to fresh sterile plates until a
pure culture was at length obtained.
Several years later Chodat and Grintzesco (9) reported that
by essentially the same method, Oocystis elliptica, Didyo-
sphcerium pulchellum , Kirchneriella lunaris, Rhaphidium poly^
morphum, Pediastrum tetras, Scenedes7nus acutus, Pleurococcus
vulgaris, Hcematococcus lacustris, and Chlorella vulgaris had
been obtained in pure culture. In cases where the number of
algal individuals is small, but the bacteria and fungi relatively
abundant, the authors point out the desirabihty of first increas-
ing the number of the former by introducing the mixture
into a mineral nutrient solution favorable for the erowth of
the alffse but not so for the fun
forms
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 27
concerned, the authors state that it is necessary to begin with
the zoospore; as a pure culture from filaments is extremely
difficult to obtain. My own experience does not bear out this
statement in all cases as it was found that especially among
the Ulotrichales pure colonies were regularly and easily obtained
from filaments.
Artari in 1902 (2) reports the isolation of Chlorococcum
infusionum and Scenedesmus caudatus in pure culture. Chick
(7) attempted to isolate Chlorella pyrenoidosa through the use
of sterihzing agents such as hydrogen peroxide and sunHght.
These trials, however, did not prove successful, as the alga
failed to show a resistance sufficiently greater than that of
the bacteria to make possible a successful separation. The
isolation was finally attained by placing a few drops of water
containing the organism on a sterile synthetic agar plate, and
spreading the same over the surface with a brush. The same
brush was used to distribute sterile water drops over the surface
of other plates, no additional algal material being added.
From the later dilutions pure colonies were obtained. Frank
(10) was unable to obtain pure cultures of Chlamydomonas
tingens by the agar plate method.
Jacobsen (11) reports the isolation of Chlorogonium and
Polytoma in pure culture. This author made use of an inter-
esting method of separation of algal species based on their
different degrees of resistance to drying. Discs of filter paper,
on which drops of water containing Spondylomorum and
Chlamydomonas variahilis had been placed, were dried in an
incubator at 28°C. After twenty-four hours, the discs were
placed in a suitable medium, but only the Chlamydomonas
species developed, Spondylomorum having been killed. Chlo-
rogonium euchlorum and Polytoma uvella also showed themselves
very sensitive to drying, whereas Chlamydomonas usually sur-
vived the desiccation. Old cultures of Chlorogonium euchlorum
proved to be very resistant owing to the presence of zygospores
which had been formed by the conjugation of gametes.
While reference might be made to a number of other inves-
tigations which deal in an incidental way with pure culture
technique, it is believed that those given will serve to indicate,
in a general way, the present status of the subject. (For further
28 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
[Vol. 1
information the reader is referred to Moore (17), Richter (21),
Kiister (14), and others.) It is apparent that the large majority
of forms isolated in pure culture belong to the Protococcales.
Only a few of the filamentous forms, several diatoms, and but
one or two species of the blue-green algse have thus far yielded
to pure culture technique.
Pure Culture Technique
n ^
GENERAL
Alga?, generally speaking, are provided with a more or less
highly developed exterior mucilaginous investment which may
be either a distinct, separable sheath, as in many of the Cyano-
phycecB, or merely a gelatinization resulting either from a
modification of the external portion of the membrane, or from
an internal secretion, as in some of the desmids. In general,
also, algse are slow growing as compared with many fungi.
In these two characteristics most of the difficulties encoun-
tered in pure culture technique among the algse find their
explanation.
Among the fungi, spores with non-gelatinous walls are readily
obtainable in a majority of the forms, and usually in great
r
abundance. When such spores are plated in the way ordin-
arily employed in bacteriological technique, a large number
of colonies free from bacteria are usually obtained. Among
the algse, however, such non-gelatinous, resistant, spores are,
if produced at all, generally present only in small quantities.
When vegetative algal cells are plated on a suitable medium,
algal colonies will often be obtained, but they usually form the
nucleus of a larger bacterial colony which has developed from
the bacteria adhering to the gelatinous surface of the algal cell.
Among those fungi in which spores are not readily obtained,
an isolation in pure culture may frequently be effected by
allowing the fungus to grow on a suitable medium until the
hyphse have outstripped the bacteria in their growth, at which
time pure mycelial transfers may be made from the terminal
portions. If, however, a hke procedure is attempted with
the algse it will usually be found that the bacteria adhere tena-
ciously to the surface of the growing filaments and are carried
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALGiE 29
along by the lengthening filaments. Except in rare cases,
nothing is to be gained by this procedure in the algae. The
task of isolating pure cultures of algse, therefore, becomes an
individual problem for almost every species as it necessitates
at once the determination of the period in the Hfe history of
any form at which the cells are free from bacteria or at which
time the bacteria can be removed by one means or another.
Having found a stage in which the alga is bacteria-free, it is
of importance next to be able to bring about this stage
more or less at will in order that the alga may be utihzed when
available. To obtain the above preliminary information, noth-
ing is more serviceable than the usual plating method on a
suitable medium.
The Medium. — The requirements of a suitable solid medium
for algal isolating purposes are, that it remain Hquid down to a
temperature at which deUcate algal cells are not injured; that
it be suitable for the growth of alg«, and as unfavorable as
possible for the growth of bacteria and fungi. For this purpose
nothing was found so serviceable as the following, the mineral in-
gredients being in the proportions recommended by Moore (17) :
Agar 10.0 grams
NH4 NO3 0.5 gram
MgS04. 7H2O . 2 gram
K2HPO4 . 2 gram
CaCb 0.1 gram
FeS04 trace
Dist. H2O 1000 CO.
The agar should be carefully washed, first in a stream of tap
water and then in distilled water, as pointed out by Richter (20).
An agar so prepared will remain Hquid down to about 34.5-
35°C., and experience has shown that even the most delicate
algal cells are uninjured by the short exposure to this temper-
ature necessary in the plating process. From six to eight cc.
of agar in a Petri dish eight cm. in diameter is a suitable quan-
tity with which to plate. Larger quantities so thicken the layer
of agar in the dish that the higher powers of the microscope,
with their objectives of short focal length, cannot be used in
locating small developing colonies.
30 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
Material to he Plated, — The alga to be plated should be
collected with as little adhering foreign matter as possible.
If it is a filamentous form which can be manipulated with a
platinum needle, it can be materially cleansed by washing
in sterilized nutrient solution such as is used in the preparation
of the agar. If the alga is a unicellular form, little can be done
in the way of preliminary cleansing. Dilutions are made in
the usual manner, the degree depending upon the number
of algal organisms present. The degree of dilution will depend
in part, also, upon the number of bacteria and fungi present
as determined by microscopic examination. It must be re-
membered that the algse grow more slowly than most bacteria
and fungi, and that unless the dilution, from the standpoint of
the total number of organisms present, is great enough, the
spread of bacterial and fungal colonies may be so great as to
make the transfer of the later-appearing algal colonies impos-
sible without contamination.
The material should be introduced into the tube of Uquid
agar while the latter is still a few degrees above its congealing
point, in order that the inoculated tube may be vigorously
shaken for some time before its contents are poured into the
Petri dish. In this way the algal cells are freed of large numbers
of either accidentally or regularly adhering bacteria.
Incubation and Transference. — The plates, after the agar has
solidified, should be turned upside down in order to prevent
the moisture which condenses on the cover from dropping,
and spreading bacteria over the surface of the agar. Failure
to do this often renders large numbers of platings worthless.
The most favorable place to keep plates is in the light of a
north window; and, as plates frequently remain under obser-
vation for many weeks, it is further desirable to have them in
a glass case to prevent outside contamination. In general it is
not advisable to cover the plates with bell jars, as it increases
the humidity in the Petri dishes and accelerates the growth
of moulds present as contaminations. The plates should be
examined frequently and when rapidly spreading colonies of
fungi or bacteria appear, these should be dissected out in order
to save the remainder of the plate.
The length of time necessary for the appearance of the algal
19141
SCHRAMM — PUEE CULTURE
31
colonies varies greatly with the species, from one to three or
four weeks usually being required, depending upon the particular
form. In most cases it is not possible to wait until the algal
colonies can be seen macroscopically because spreading bacterial
and fungal colonies usually encroach on the former to such
an extent that a pure transfer is no longer possible. It becomes
necessary, therefore, to look the plates over from time to time
with the compound microscope in order to locate algal colonies in
very earlystages of development. For this purpose a 12 mm. ob-
jective is extremely serviceable, as its focal length is of sufficient
magnitude to enable one to use it through the agar layer and
glass bottom of a Petri dish and at the same time obtain a
magnification considerably greater than that afforded by the
ordinary low-power obj ective. The colonies located are conven-
iently marked by placing a small ink dot directly opposite them
on the bottom of the Petri dish. Transfers should be made to
agar slants by means of a minute platinum-foil spatula with which
the agar directly over the ink dot can be neatly dissected out
and transferred to the slant. It is not possible, in most cases, to
make successful transfers with a platinum needle because the
algal colony is usually composed of firmly cohering cells and,
even in repeated attempts, not a single individual will adhere to
the needle. Since many of the colonies are in the deeper strata,
it is well to spread out the transferred agar fragment in a
thin sheet in order to expose the contained algal cells directly
to the air. Unless this is done, subsequent development in
the slant may be extremely slow. Although bacteria grow
slowly on this synthetic agar, their development is usually
sufficient in a week to indicate whether the transfer has been
successful or not. The purity of the culture may be further
tested by making transfers to media more suitable for bacterial
growth.
With this brief preliminary consideration of some of the more
general phases of pure culture technique in the algse, the isola-
tion of single species will now be considered and attention called
to the special problems and the technique involved in their
isolation.
2
[Vol. 1
32 ANNALS OF THE MISSOURI BOTANICAL GARDEN
SPECIFIC
CHLOROFHYCE^
Chlamydomonas pisiformis Dill forma minor Spargo. —
Chlamydomonas species frequently occur in water rich in organic
materials, and teeming with bacteria. When the alga was in the
resting condition, the mucilaginous cell walls were found so
impregnated with bacteria as to render isolation in pure culture
impossible. Platings with motile cells, however, showed that
the latter were absolutely free from regularly adhering bacteria,
but the number of bacteria present rendered the plates worth-
I
less. Then the gelatinous masses of resting cells were repeat-
edly washed with sterile water and finally placed in distilled
water where, after twelve to twenty-four hours, zoospores
appeared in great abundance and congregated on the side
of the vessel nearest to the light. A minute portion of
this liquid containing the zoospores was removed with a fine
capillary tube and introduced into a tube of liquid agar and
platecL In platings thus made, numerous colonies of Chlamydo-
monas appeared and the number of bacterial colonies was so
r
small that a large number of successful pure transfers were made.
Where the number of available motile cells is small and it
is important that isolations be made from these, a modification
of the method used by Barber (3) in the isolation of yeasts
> ■
and bacteria was frequently used to advantage. A large num-
ber of small, capillary pipettes were made and sterilized. After
locating the cell or cells desired, they were removed with a
pipette while being observed under the microscope, and trans-
ferred to a drop of sterile nutrient solution or water. This
process was repeated until it was certain that the number of
bacteria had been reduced sufficiently to admit of successful
plating. They w^ere then taken up again by means of a sterile
pipette, transferred to a tube of liquid agar, and plated.
Numerous pure cultures were obtained in this way.
Stichococcus hacillaris Nag., and S. suhtilis (Ktitz.) Klercker.
Preliminary platings with these forms showed that the cells, as
obtained from the soil, yielded abundant bacteria-free colonies,
and the problem of isolation became one of merely obtaining
clean material and diluting sujfficiently. Both of these species
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 33
of Stichococcus are soil-inhabiting and can be obtained — practi-
cally free from other algae — on flower pots and greenhouse soils.
The former species, because of its minute cells and the readiness
with which the filaments resolve themselves completely into
their constituent cells when placed in water, is a particularly
easy one to obtain in pure culture. Rich material may be
diluted until plates obtained from it show a sufficiently small
number of bacterial colonies to admit of pure transfers and
yet enough algal colonies for a number of transfers. S. suhtilis
is a larger species and the cells remain attached in rather long
filaments. However, with vigorous shaking and previous
teasing apart with needles, a sufficient number of single cells
and small fragments of filaments are introduced to make
possible numerous successful isolations. The washing of the
cells to remove adhering bacteria can, in these species and
many others, be largely accomplished by introducing the
raw material into test-tubes containing sterile mineral nutrient
solution or water, stoppering, and shaking vigorously. Direct
transfers from these to liquid agar, or to tubes of sterile water
for further dilution, may then be made. This procedure fre-
quently enables one to make successful platings where the
direct transfer of raw material to liquid agar results in constant
failure.
Chlorella vulgaris Bey., and Chlorella sp.- — Both of these
species were isolated from soil in the open. An exterior gelat-
inous investment is, as in the two above mentioned species
of Stichococcus, conspicuously absent, and preliminary experi-
ments demonstrated that a large number of the vegetative
cells were freed from all accidentally adhering bacteria by being
shaken in the liquid agar before plating. The problem of
isolating these species again becomes one of clean material
and sufficient dilution. Species of Chlorella are perhaps the
among the
the
requiring little more than a direct application of bacteriological
methods.
Attention should be called to another method — really a
modification of the one just given — by means of which Chlorella
species may be obtained in pure culture. Its application is
not necessary in the species of Chlorella investigated^ since
[Vol. 1
34 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the vegetative cells can be so readily freed from adhering
bacteria. But its general applicability to other forms justifies
its mention at this place. Chlorella, like many other genera
of the Protococcales, forms non-motile endogenous daughter
cells which remain enclosed in the mother wall for varying
lengths of time. The enclosed daughter cells are in all cases
free from adhering bacteria. A group of daughter cells still
enclosed within the mother membrane may be removed by
means of a capillary pipette to a drop of sterile water, and from
here to a succession of others until all readily removable bacteria
have been left behind. The last transfer should be made to
a drop of sterile water on a small sterile cover glass. By a
slight pressure of a second cover glass, the mother membrane
may be ruptured, liberating the enclosed, bacteria-free cells.
The two cover glasses should then be introduced into a tube
of liquid agar, the latter shaken vigorously, and finally poured
into a Petri dish. Frequent isolations have been made in this
way, and its importance in forms whose vegetative cells cannot
be freed from adhering bacteria, and which do not form motile
spores but only non-motile endogenous daughter cells, can
hardly be overestimated.
Pleurococcus vulgaris Menegh. — The majority of PZewrococcus
cells, when thoroughly washed, will be found free from bacteria.
A difficulty which frequently arises is that the alga grows so
very slowly that fungi — which are persistently present in Pleu-
rococcus cultures — take entire possession of the plates before
a transfer can be effected. But with careful searching, minute
colonies— often consisting of but a few cells — can usually be
found and successfully transferred. The transferred colony,
however, usually makes extremely slow progress in its growth
on agar. Much better results are obtained when transfers
are made to evaporimeters (as devised by Livingston (15))
supplied with the mineral nutrient solution.
Scenedesmus sp., and Kirchneriella sp. — Both of these species
were obtained in pure culture by washing and diluting clean,
concentrated material in sterile mineral nutrient ' solu-
tion, and then plating. The great majority of the colonies
of both species were contaminated with bacteria, pure colonies
being very rarely found. This fact, together with the gelatin-
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 35
ous exterior characteristic of the cells of both species,
makes it probable that the pure colonies developed, not from
mature individuals, but from autocolonies (produced within
mature cells) which either had just escaped from the mother
cell or had done so during the vigorous shaking, — in either of
which cases they are free from adhering bacteria.
Chlorococcum humicola (Nag.) Rabenh. — This species was
isolated in the zoosporic condition. The alga, collected from
soil, was placed in sterile mineral nutrient solution and after
twenty-four hours produced zoospores in abundance. Platings
with these yielded numeroils pure colonies from which successful
transfers were made. In this connection it should be men-
tioned that all zoospores thus far experimented with — including
a considerable variety of forms — have been found free from
bacteria. It is needless to say, therefore, that the presence
of zoospores in the life cycle of any alga provides a logical
point of attack for its isolation in pure culture. While not all
the attempts to isolate zoosporic forms in pure culture have
proved successful, it is entirely probable that they will when
the general technique is more closely adapted to individual
forms.
Protosiphon hotryoides (Kiitz.) Klebs. — The vegetative plant
of Protosiphon, with its root-like process extending into the
soil and the large aerial portion, is so persistently covered with
bacteria that its isolation in pure culture in this condition
is quite impossible. With slight desiccation, however, large
numbers of chlamydospores with dry non-gelatinous mem-
branes appear, which, at least so long as they remain enclosed
within the mother membrane, are free from bacteria. From
these, isolations in pure culture can be readily made according
to the second method suggested for Chlorella — by carefully
washing an individual plant filled with chlamydospores, hber-
ating the latter by teasing with needles or by a shght pressure
of the cover glass, and plating in the usual manner. Another
method which has yielded pure cultures, but which is not
to be recommended because it is far less reliable than the one
just described, is based on the use of the motile gametes.
When vigorous Protosiphon plants, growing on soil, are covered
with distilled water, gametes, which congregate in the Hghted
36 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
side of the vessel, are produced in large numbers. Plates made
with this material yield an occasional pure culture, but most of
the gametes fail to develop. It is impossible at present to say
whether the colonies develop from newly formed zj'^gotes or from
gametes which fail to conjugate.
Stigeoclonium tenue (Ag.) Kiitzing. — The ease and certainty
with which zoospores can be induced to develop in this form,
and their extreme abundance, makes it, although a filamentous
alga, an especially easy one to isolate. Freshly collected and
thoroughly washed filaments of Stigeoclonium, placed in dis-
tilled water or sterile nutrient solution, will, in from twelve to
twenty-four hours, develop a great abundance of zoospores.
Cultures prepared in this way contain so small a number of
bacteria that plates containing a hundred or more Stigeoclonium
zoospores are sufficiently free from bacterial colonies to
render numerous successful pure transfers possible. Although
a filamentous form, Stigeoclonium grows exceedingly well on
the mineral nutrient agar. While other members of the
Chcetophoracece were not experimented with, it is reasonably
certain that forms like Microthamnion, Chcetophora, and Drapar-
naldia, all of which readily yield large quantities of zoospores,
may be obtained in pure culture by a method identical with or
similar to the one employed in the isolation of Stigeoclonium.
Oedogonium sp., and Vaucheria sp. — While neither of these
forms were obtained in pure culture, the observations made
render it altogether likely that this will be possible when a little
more attention is given to the cultural solutions. Repeated trials
with the vegetative filaments demonstrated that from the
latter no pure cultures could be obtained directly. The
oospore proved equally unsatisfactory because the oogonial
wall is covered with adhering bacteria. Again, the oospore
is, in most cases, so firmly and completely united with the oogo-
nial wall that its separation from the latter is at present impos-
sible. In both forms, however, zoospores are readily obtained,
and preliminary experiments demonstrated that these, like
zoospores in general, are bacteria-free. Where zoospores could
not be obtained in large quantities, individual ones were isolated
with sterile pipettes, washed repeatedly in sterile water, and
then either plated in the usual manner, or introduced into a
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 37
tube of sterile mineral nutrient solution. Although the great
majority of such isolations remained bacteria-free, the zoospores
failed to develop, and finally died. It is only necessary, there-
fore, to find some medium in or on which the zoospores will germ-
inate and develop into plants, to effect a pure culture of
Vaucheria or Oedogonium. BulbochcBte was not used, but in
all probability this form will lend itself to a similar technique.
Conjugates. — Thus far it has not been possible to obtain a
pure culture of any member of the Conjugales. The repre-
sentatives of this order, in their vegetative phases, are provided
throughout with an exterior gelatinous investment which is
very generally impregnated with bacteria. All attempts to
obtain pure cultures from vegetative individuals failed. Fur-
ther, there is a complete absence in the order of motile spores
and, in general also, of separable, asexual, endogenous spores.
The zygospore, therefore, suggests itself as a possible means
of solving the problem, especially in those forms where it is
produced endogenously, and where it does not subsequently
coalesce with the wall of the gametangium. While pure
cultures were not obtained from these, the method used in
Spirogyra setiformis is of interest and may prove serviceable
in the ultimate isolation of these forms in pure culture.
Filaments containing mature zygospores, but in which the
zygospore-containing cell walls were still completely intact,
were washed repeatedly in sterile water and then broken up
as thoroughly as possible with needles; in this process, numer-
ous zygospores were freed from the enclosing walls, later to be
taken up with sterile pipettes, and transferred to sterile drops
of water. Each zygospore was subsequently transferred from
ten to twenty times to fresh, sterile water drops, and finally
taken up with a sterile pipette. When a considerable number
of zygospores had thus been isolated, they were introduced into
a tube containing a few cc. of sterile water, vigorously shaken,
and the entire contents poured out into a Petri dish containing
a layer of sterile nutrient agar. After rocking the dish for a
short time, it was allowed to remain quiet until the zygospores
had settled down on the surface of the agar. The free water
was then very slowly and carefully, but completely, drained
from the surface of the agar, and the plate allowed to remain
38 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
in the light. While in a few cases bacterial colonies developed
about the zygopsores, it was found that the great majority
were free from all adhering bacteria. Such zygospores as were
bacteria free were then transferred to test tubes containing
sterilized mud and pond water. Although about sixty such
transfers were made, not a single one yielded a growing culture,
although zygospores kept in battery jars in the laboratory
showed a high percentage of germination. It will require
further experiments to find a suitable medium for the germina-
tion and subsequent growth of isolated zygospores. However,
the isolation of bacteria-free zygospores justifies the opinion
that with them it will, sooner or later, be possible to culture
Spirogyra in a state of purity.
HETEROKONT.E
Botrydium granulatum (L.) Greville.— This form is, in its
general morphology, so similar to Protosiphon, that the tech-
nique, as regards the use of chlamydospores, described for the
latter, is entirely applicable here. Botrydium when submerged,
however, forms an abundance of zoospores instead of gametes,
and from these pure cultures can be obtained with great ease
when plated in the usual manner. The method for using the
chlamydospores can also be considerably abbreviated in Botryd-
ium. When the plants form chlamydospores, the aerial
globular portion of the plant collapses. The cell, however, is
so large that the aerial bag can be torn open with fine sterile
forceps, the spores removed under a hand lens with a needle
and transferred directly to liquid agar. Platings made in this
way show a very slight bacterial contamination, and pure
transfers can be made in abundance. While a direct, bacteria-
free transfer has not been thus effected, it is altogether probable
that it can be done. The pure transfers of Botrydium having
been obtained, it was found that their development on agar
was extremely slow, and ultimately all of the cultures died.
Further experiments will be necessary in order to provide a
favorable medium for growth. The clay-cup evaporimeter
may perhaps prove of service in this connection as it did in
the case of Pleurococcus,
Botrydiopsis sp.— This form was found abundantly during
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 39
one season on soil in the greenhouses. The vegetative cells
when placed in water readily produce zoospores, and isolations
were made from these with little difficulty. Unlike Botrydium,
this form grows exceedingly w^ell on the mineral nutrient agar.
BACILLARIALES
The diatoms were encountered onlj'- incidentally in connec-
tion with other forms, and no particular effort was made to
isolate forms in pure culture. Although diatoms, in general,
have a gelatinous exterior, a small Navicula was on several
occasions obtained in pure culture and grown successfully.
It should be said, however, that the great majority of diatom
colonies obtained were contaminated with bacteria.
CYANOPHYCE^
In the class Cyanophycece, the most difficult problems of
isolation are met. The almost universal presence of an abun-
dance of external mucilaginous material, the complete ab-
sence of ciliated reproductive cells, and the virtually complete
absence of free, endogenous spores, renders the technique
particularly difficult. The gelatinous investments are, in
all cases investigated, impregnated with bacteria which can-
not be completely removed by the most vigorous washing.
Among the forms studied were Aphanocapsa, several species
each of Oscillatoria, Nostoc, and Anabcena, Cylindrospermum,
and Microcoleus. Of these, only two species, one of Oscilla-
toria and one of Microcoleus, were obtained in pure culture.
In the isolation of these two forms, silicic acid j ally was found
to be indispensable. While directions for preparing this
medium are to be found in many places in the literature, certain
difficulties encountered in its preparation have made it desirable
to give at this time, and in some detail, the method used.
As regards the preparation and mixing of the sodium silicate
and hydrochloric acid solutions, the directions given by Smith
(22) may be followed. It is only necessary to point out in
this connection that if Merck's "sodium silicate pure crystals"
is used, the solution should be made up with cold water. If
hot water is used, an unidentified substance (insoluble in cold
water) goes into solution, and frequently causes the coagulation
r
P
[Vol. 1
40 ANNALS OF THE MISSOURI BOTANICAL GARDEN
of the silicic acid-hydrochloric acid mixture before dialysis is
complete. A point of very great importance is the preparation
of the collodion dialyzing bags. As has been pointed out by
Kellerman (12)^ and others, the degree of permeability of the
bags depends, in a large degree, upon the way in which they are
made. If the guncotton solvent is made from equal parts of ether
and absolute alcohol, the bags will, in most cases, have a very low
permeability, and coagulation of the enclosed silicic acid solution
will frequently result before dialysis is complete. The degree
of impermeability is further increased by drying the bags
rapidly. If, however, 95 per cent (instead of absolute) alcohol
is used, and the bags are allowed to dry spontaneously by invert-
ing the test-tubes in which the bags are being prepared in
suspended wire baskets, a much higher degree of permeability
will be obtained.
Bags prepared with 95 per cent alcohol were used, and the
silicic acid-hydrochloric acid mixture dialyzed in tap water
until the chloride content was no greater than that of the water.
The silicic acid solution was further purified by dialyzing in
changes of ordinary distilled water and finally in triply distilled,
nitrogen-free water. In this extended dialysis, a considerable
portion of the silicic acid is lost, and it usually becomes necessary
to concentrate the solution to obtain a jelly of sufficient firm-
ness. This is best carried out in heavy, two-liter suction-flasks
in which the pressure is reduced until the solution boils at
from 35 to 40°C. If the concentration is carried out at higher
temperatures, coagulation sometimes results. In order to pre-
vent the violent bumping which always takes place unless some
special precautions are taken, it is only necessary to bring
through the rubber stopper at the top of the suction-flask a
glass tube drawn out at the bottom to a very fine capillary,
which dips into the solution. The top of this tube, outside of
the rubber stopper, should be provided with a piece of rubber
tubing and pinch cock to regulate the intake of air. The air
thus admitted may first be washed to remove carbon dioxide,
ammonia, or other impurities. The concentration should be
continued until a sample, when congealed, has the proper
consistency. The directions given by Smith (22) for coagu-
lating the medium apply here and it need only be mentioned
1914]
SCHBAMM — PURE CULTURE METHODS IN THE ALG^ 41
that the concentration of the mineral nutrients employed in
the agar, 0.1 per cent, is quite sufficient to bring about coagu-
lation.
After it had become probable that no blue-green alga, in
the ordinary vegetative condition, could be isolated by the
usual plating method, tubes containing from two to three
inches of solid, sterile, synthetic agar were inoculated at the
surface with a species of scillatoria . The tubes were then
completely wrapped in black paper, leaving only the very
bottom exposed to the light, and inverted. It was hoped that
in the rapid growth of the alga through the agar, the bacteria
might be left behind. The growth toward the light in some
cases amounted to eight mm., and more, per day. When the
growth had approximately reached the bottom of the tube,
the end of the latter was broken away, the surface of the agar
seared, and transfers made from the interior of the agar plug.
Although the experiment was repeated many times, and a
total of at least fifty transfers made, a pure culture was never
obtained, bacteria always being present. Large Petri dishes,
containing a layer of sterile synthetic agar, were then inoculated
at one edge with a species of scillatoria , and the dishes so
placed that the point of inoculation was farthest away from
the light. The alga grew rapidly (on the surface of the agar)
toward the light, and just before reaching the opposite edge of
the dish, transfers were made from the farthest advanced
filaments. Although transfers to fresh agar surfaces were
continued to the number of six, a pure culture was never
obtained.
The experiment was then repeated, surfaces of silicic acid
jelly replacing those of agar, with the result that numerous
pure transfers were obtained from the second plate. A species
of Microcoleus was obtained in pure culture in an identical
manner.
Most members of the scillatoriacece are provided with a
sharply delimited, gelatinous sheath. Reproduction is effected
by the formation of hormogonia which glide out of the sheath,
move about slowly for a time, and then come to rest. In
forms like Microcoleus, Lynghya, and some species of Oscilla-
ioria in which the hormogonia escape from definite sheaths,
42 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
leaving the latter behind, it is fairly certain that the hormo-
gonium is originally free from bacteria, but becomes contam-
inated in passing through the older portion of the empty sheath
and out of its terminal opening, both of which are more or less
infected with bacteria. The persistence with which the bacteria
chng to the hormogonium of Oscillatoria, once having infected
it, is clearly shown by cultures on agar surfaces. Although
a single hormogonium may have moved as much as two inches
away from its parent filament, creeping all the while over a
sterile agar surface, the hormogonium will be found covered
with bacteria, and the path over which it moved will be clearly
indicated by a continuous, linear colony of bacteria. With
the use of silicic acid jelly, however, the multiplication of the
bacteria is reduced to such an extent that, after a time, hormo-
gonia escape uncontaminated, and begin the development of
pure colonies. Transfers from these, however, grow very
slowly and in most cases eventually die. It seems probable,
when Oscillatoria and Microcoleus have been completely sepa-
rated from the invariably present bacteria, that the media
which were favorable in the presence of the bacteria, become
unfavorable in their absence. Further work will be necessary
to grow these forms successfully after they have been isolated
in pure form. The silicic acid jelly method was also attempted
with the above mentioned heterocystic forms; however, up
to the present time, no successful isolations have been made.
Discussion
It is apparent that the technique involved in the isolations
just referred to depends entirely on mechanical separation
of one kind or another. This method is reasonably efficient
in those species in which zoospores or other free endogenous
spores are readily obtainable, or in which vegetative cells are
either free from bacteria or can be rendered so by mechanical
means. It is true that even in some species forming free
endogenous spores, the above methods have not yielded pure
cultures, as, for instance, in Vaucheria, Oedogonimn, and Spiro-
gyra. In these cases, however, it should be pointed out that
it is not the isolation technique which is at fault but rather
1914]
SCHRAMM — PURE CULTURE METHODS IN THE ALG^ 43
r
the cultural methods. Zoospores and zygospores, respectively,
free from other organisms, were obtained in these cases but
failed to develop in the cultural media subsequently supplied.
There can be little doubt, however, that the latter dijfficulty
will be overcome in time.
Except in the Oscillatoriacece, little progress was made in
the Cyanophycece. The problem appears especially difficult
in the Coccogoneales where all forms of motile reproductive
bodies are absent, and in which the vegetative cells apparently
cannot be rendered free from adhering organisms by mechan-
lica means. Even in the heterocystic Hormogoneales, the
situation is a difficult one, the more slowly moving hormogonia
apparently being unable to escape the bacteria.^ While no
experiments were made along these lines, it appears highly
desirable to attack the problem in the latter group through
the spore. It is well known that the spores of blue-green
algse are extremely resistant to heat, and it does not appear
improbable that the bacteria — especially if they are all in the
vegetative condition — could be killed by heat, leaving the
algal spores unharmed. Chemical sterilizing agents may also
■
prove of value here. The latter may also prove serviceable
with members of the Coccogoneales and certain of the grass -
green algse which have thus far failed to yield to the technique
employed.
Conclusions
1. By adapting methods of pure culture technique to indi-
vidual species of algse, it has been possible to isolate in pure
culture the following forms:
ChlorophycecB. — Chlamydomonas pisiformis Dill forma minor
Spargo, Stichococcus hacillaris Nag., >S. subtilis (Ktitz.) Klercker,
Ulothrix sp., Chlorella vulgaris Bey., Chlorella sp., Pleurococcus
vulgaris, Scenedesmus sp., Kirchneriella sp., Chlor ococcum humi-
cola (Nag.) Rabenh., Protosiphon hotryoides (Kiitz.) Klebs,
Stigeoclonium tenue (Ag.) Kutzing, and a number of others
of uncertain identity.
'In a contribution which has just appeared (Kulturversuche mit Chlorophyll-
fahrenden Mikroorganismen, III. Zur Physiologie der Schizophyceen. Beitr. z. Biol,
d. Pflanzen 12: 49-108. 1913), Ernest G. Pringsheim reports the isolation in pure
culture of a species of Nosloc. The method used was that of repeated transfers to
sterile silicic acid jelly plates.
44 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
I Vol. 1
H eter okontce . — Botrydium granulatum (L.) Greville, and Botry-
diopsis sp.
B acillariales . — Navicula sp,
Cyanophycece — Oscillatoria sp., and Microcoleus sp,
2. In addition, zoospores from Vauchena and Oedogonium,
and zygospores from Spirogyra have been isolated free from
other organisms.
In conclusion, the author wishes to express his gratitude to
Dr. Geo. T. Moore, at whose suggestion the work reported herein
was undertaken, whose advice and interest have been a source
of constant help; and to Mildred Spargo Schramm, for kindly
many
Literature Cited
1
Artari, Alexander, Ueber die Entwicklung der griinen Algen unter Ausschluss
" ' ' A^ssimilation. Bull, de la Soc. Imp. de
der Bedingu
39-47. 1899.
2.
deut. bot. Ges. 20: 172-175. 1902.
griinen
8. Barber, M. A. On heredity in certain micro-organisms. Kansas Univ. Sci^
Bui. 4: 3-48. 1907.
4. Beyerinck, M. W. Kulturversuchc mit Zoochlorellen, Lichengonidien und anderen
niederen Algen. Bot. Zeit. 48: 725-785. 1890.
5- , Bericht iiber meine Kulturen niederer Algen auf Nahrgclatine.
Centralbl. f . Bakt. 13 : 368-373. 1893.
6- , Over gelatineculturen van eenccllige groenwicren. [Reviewed in Cen-.
tralbl. f. Bakt. 8: 460-462. 1890.]
7. Cluck, Harriette. A study of a unicellular green alga, occurring in polluted water,
with especial reference to its nitrogenous metabolism. Proc. Roy. Soc. 71:
458-476. 1903.
8. Chodat, R., et Goldflus, M. Note sur la culture des Cyanophycdcs ct sur le dcvel-,
oppment d'Oscillatori^es coccog^nes. Bull, de 1' Herb. Boissier 5: 953-959.
1897.
P
9. Chodat, R., et Grintzesco, I. Sur Ics mcthodes de culture pure des algucs vcrtes.
Frank, Theodor, Kultur und chemischc ]
tingens. Bot. Zeit. 62: 153-188. 1904
1900
11. Jacobsen, H. C. Kulturversuche mit einigen niederen Volvocaceen. Zeitschr.
f. Bot. 2: 145-188. 1910.
12. Kellerman, K. F. The permeability of collodion tubes. Centralbl. f. Bakt,
11.34: 56-60, 1912.
Kriiger, Wilhehn, Beitrage zu
Schleimflusscs) der Laubbaxma
4: 69-116. 1894.
smen
r
Ernst, Anleitung zur Kultur der Mikroorgam*smen. 105-107, 1907
1914]
SCHRAMM— PURE CULTURE METHODS IN THE ALG^ 45
15. Livingston, B. E. A new method for cultures of algse and mosses. Plant
World II : 183-184. 1908.
16. Miquel, P. De la culture artificielle des Diatom^es. Le Diatomiste 8: 73-75,
1892; 9: 93-99. 1892. [Reviewed in Compt. Rend. 114: 780-82. 1892.]
17. Moore, G. T. Methods for growing pure cultures of algse. Joum. Appl.
Microsc. 6: 2309-14. 1903.
18. Ogata, M. Ueber die Reinkultur gewisser Protozoen (Inf usorien) . Centralbl.
f. Bakt. 14: 165-169, 1893.
19. Radias, M. Sur la culture des algues h Petat de purity, Actes du Congr^s Int,
de Botanique, Paris, 1900, 163-167.
20. Richter, Oswald, Reinkulturen von Diatomeen. Ber. d. deut. hot. Ges. 21:
493-506. 1903,
21. , Die Emahrung der Algen. Monograph, u. Abhandl. z. intemat.
Revue der ges. Hydrobiol. u. Hydrograph. 2: 31. 1911.
22. Smith, E. F. Bacteria in relation to plant diseases. Publ, Carnegie Inst. 27":
37-39. 1905.
23. Tischutkin, N. Ueber Agar-Agarkulturen einiger Algen und Amoben. Cen-
tralbl. f. Bakt. 11. 3: 183-188. 1897.
24. Ward, H. Marshall, Some methods for use in the culture of algse. Ann. Bot.
13:563-566. 1899,
Graduate Laboratory, Missouri Botanical Garden.
THE IDENTIFICATION OF THE MOST CHARACTER-
ISTIC SALIVARY ORGANISM, AND ITS RELATION
TO THE POLLUTION OF AIR^
AUGUST G. NOLTE
Introduction
Bacteriologists and sanitary engineers have, within the last
score of years, given much attention to the detection of excre-
mental pollution in water. They have shown that by making
it possible to recognize certain characteristic accompanying
organisms, bacteriological methods are capable of revealing
this kind of pollution even when it exists to such a small degree
as to be beyond the
of chemical detection. Small
these quantities of contaminating substances may seem, they
may nevertheless endanger the health of a whole community
by exposing it to possible pathogenic organisms derived from
the excreta of a diseased host.
It is not merely by the aggregate bacterial yield that the
potability of a water in its relationship to disease is judged
but more specifically by the species
of bacteria
and
their relative abundance. The micro-organisms which serve
index of
as
examination
made
and for which special quantitative
the members of the colon group.
These, from their
and
abundance
are characteristic of material of excremental origin. Their
presence in water in sufficient quantity indicates pollution,
and their relative abundance serves as an index to the extent
of the latter.
Bacteriological technique has not as yet been applied to
the same extent in the detection of pollution in air. Chemistry
has, up to the present time, been of more practical value here.
1 An investigation carried out at the Missouri Botanical Garden in the Graduate
Laboratory of the Henry Shaw School of Botany of Washington University, and
submitted as a thesis in partial fulfillment of the requirements for the degree of
master of arts in the Henry Shaw School of Botany of Washington University.
Ann. Mo. Bot. Gaed., Vol. 1, 1914
i
(47)
48 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
The proportion of carbon dioxide is still the standard mainly
relied on for estimating pollution of air by materials given off
from the human body, although it is recognized that other
factors may be of more importance. This method of exam-
ining air, however, is of little or no value in furnishing an index
to the probable or possible contamination with disease-pro-
ducing germs, for there is at present no reason for believing
that such organisms are given off in the breath during ordinary
quiet breathing. Thus, M. H. Gordon^ calls attention to the
following: Tyndall observed that expired air is optically purer
than inspired air; Cornet found air expired by tubercular patients
to be free from the tubercle bacillus; and Straus has shown that
expired air is not only comparatively free from bacteria, but
that it is considerably purer in this respect that inspired air.
It nevertheless appears probable that bacteriology rather than
chemistry will furnish a means of investigating the pollution
of air by disease-producing germs. The problem at hand is
to devise, if possible, a method for estimating the degree of
pollution of air by pathogenic organisms (given off from the
human body) in a manner similar to that employed in estimat-
ing the extent of pollution of water by similar organisms of
excremental origin.
HiSTOKICAL
L
It appears that the present status of bacteriological analysis
of air is comparable to that of bacteriological analysis of water
some years ago, when the total number of bacteria in a given
quantity was the chief factor determined. There are various
ways in which pathogenic organisms may gain access to the
air and ultimately to another individual. In addition to trans-
fer by direct contact, disease-producing organisms may be given
off in the urine, in feces, in sputum, or from the surface of the
skin. Recently, also, attention has been called to the possi-
biUty of the pollution of air by the scattering of fine particles
of mucus and saliva from the mouth in the acts of coughing,
sneezing, and loud speaking. The latter methods of air pollu-
J Report on a bacterial test for estimating pollution of air. Supplement to the
Thirty-second Annual Report of the Local Government Board (London), containing
the Report of the Medical Officer for 1902-3. 421-471. 1904.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 49
tion are the ones to be considered in this investigation. They
doubtlessly constitute an important means whereby patho-
genic organisms enter the air from an infected person, subse-
quently to be transmitted to other individuals.
The discharge of sputum furnishes the most obvious way
whereby pathogenic organisms may be expelled from the mouth.
The expectorated mucus dries, and, in the form of dust, may
later be inhaled to produce infection. The work of Flugge
and members of his school/ however, has drawn attention
to a more direct and no less important way by which germs may
be aerially conveyed from the mouth. The problem of trans-
mission of micro-organisms by means of particles of mucus
expelled from the mouth in various expiratory acts, was attacked
in two ways by the investigators referred to above: 1. The
mouth was artificially infected with a culture of Bacillus pro-
digiosus. This organism was chosen because the red pigmenta-
tion of the colonies renders the identification easy. After agar
plates had been placed at various distances from the person
r
being experimented upon, the individual proceeded to speak,
cough, sneeze, etc. At the end of the experiment the agar
plates were covered and incubated at 25° C. for 3 days, during
which time the characteristic red colonies of B. prodigiosus
made their appearance. The possibility of error due to the
previous presence of this organism in the air of the room was
excluded by exposing a series of agar plates immediately before
the experiment began, with the result that in all cases ' the
organism failed to appear. The length of time that droplets
of mucus remained suspended in the air after the several expir-
4
atory acts was determined by exposing plates at various periods
after the experiment had been completed. 2. Glass slides or
empty Petri dishes were placed at various distances from a
tubercular patient. The droplets of mucus expelled during
coughing, and deposited upon the glass slides, etc., were either
examined microscopically or were washed off and injected
intraperitoneally into guinea-pigs. In the former case a bacillus
4
giving the characteristic staining reaction of the tubercle
bacillus was found, and in the latter the development of tuber-
» Gordon, loc. at
[Vol. 1
50 ANNALS OF THE MISSOURI BOTANICAL GARDEN
culosis in the inoculated animals resulted. In other experiments;
guinea-pigs, instead of being inoculated, were directly exposed
to the coughing of tubercular patients with the result that a
number of the animals so exposed contracted tuberculosis.
Varied and repeated experiments along these lines established
the fact that in the acts of coughing, sneezing, and loud speak-
ing, fine droplets of mucus are ejected into the air, that they
float about and may be wafted by air currents, such as obtain
in ordinary rooms, to a distance of from 24 to 40 feet.
The most thorough investigation in recent years of the
problem of air pollution with micro-organisms was made by
Dr. M. H. Gordon. 1 This author believed that the positive
recognition of disseminated saliva constituted an important
step in the development of an applicable bacteriological method
for the examination of air. By bacterial analyses of a number
of samples of saliva obtained from normal individuals, Dr.
Gordon determined that the streptococci are the organisms
most abundantly present in saliva. Of these he was able to
differentiate four morphologically different types— longus , me-
dius, hrems, and conglomer atus . In endeavoring to differentiate
these organisms on a physiological basis a study was made of
their virulence, relation to oxygen, optimum growth temper-
ature, pigment production, motihty, gelatin liquefying power,
indol production, action on litmus milk at ST'^C., and action on
various carbohydrates.
It was found that the micro-organism which is most useful
in the detection of droplets of saliva is Streptococcus hrevis
because it is the only one among the salivary cocci found which
changes the color of neutral red broth to yellowish green, and
produces acid and clot in milk. Having developed a means
of differentiating the coccus most characteristic of saliva, Gordon
next examined the open air for the presence of micro-organisms
characteristic of saliva. In these experiments broth plates were
exposed for a stated length of time and incubated anaerobically
at 37^0. In but very few cases were the organisms isolated
from the air.
A further means of differentiating the characteristic salivary
» Loc. cit.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 51
coccus from the air cocci was sought in the action of the two on
various organic substances. In this capacity the several broths
containing lactose, syringin, and coniferin, proved es^pecially
serviceable. In lactose broth the typical salivary coccus was
positive, i. e., it produced acid, whereas the air cocci were
negative. In the syringin and coniferin broths, the air cocci
were positive, the typical salivary coccus negative.
L
To determine whether or not particles of saliva were dis-
seminated through the air during the acts of coughing, sneezing,
and loud speaking, Gordon performed experiments in a large
and in a small room, using, at first, Fliigge's method of arti-
ficially infecting the mouth with a living culture of Bacillus
prodigiosus and placing sterile agar plates at various distances
in front of and behind the speaker. After |-1 hour of loud
speaking, it was found that B. prodigiosus had been disseminated
to a distance of 40 feet in front of and of 12 feet behind the
speaker. In other experiments in which no artificial infection
of the mouth was resorted to, but in which the characteristic
salivary coccus served as the index of dissemination, it was
found that after f-1 hour of loud speaking Streptococcus hrevis
appeared on broth plates placed as many as 12 feet in front of
and behind the speaker. In similar experiments in which
speaking was continued for one hour in an ordinary conver-
sational tone, no dissemination of the salivary Streptococcus
m
could be detected.
From his experiments Dr. Gordon inferred that there were
certain streptococci normally present in saliva which are appli-
cable for the detection of droplets of saliva in air in much the
same manner that Bacillus coli can be applied for the detection
of fecal matter in water.
*
The Identification of the Most Characteristic Salivary
Organism
With a view of determining the organism most characteristic
of saliva, I have undertaken, as a first step, a bacteriological
analysis of the saliva of a normal individual. In this examina-
tion special attention was paid to the type of organism most
abundantly present. Having determined the type, i.e., whether
bacillus, coccus, or spirillum, characteristic reactions for it
were next sought in order to render its recognition easy. Since
[Vol. 1
52 ANNALS OF THE MISSOURI BOTANICAL GARDEN
a possible relation of the characteristic salivary organism to
the pollution of air was to be investigated, it was necessary
to examine the outdoor air free from human contamination
for the presence of micro-organisms closely allied to those
characteristic of saliva. As particles shed from the skin may
^be present in the air, it was further necessary to examine those
micro-organisms found on the skin which were closely allied
to the ones characteristic of saliva.
■ In examining the saliva for the type of micro-organism
most constantly present, i.e., whether bacillus, coccus, or
spirillum, the dilution method was used. It is reasonably
safe to assume, after repeated trials, that the type of micro-
organism which persists longest in continued dilutions is the
type most abundant in the material examined. This is true
provided the medium on which the organism is grown is approx-
imately equally favorable for the development of all the types '
present. The dilutions were carried out as follows: A sample
of saliva was collected in a sterile test-tube and 1 cc. introduced
into a second tube containing 9 cc. of sterile distilled water.
The contents of the latter were then thoroughly mixed and
I cc. of the Hquid introduced into a third tube likewise con-
taining 9 cc. of sterile distilled water. This procedure was
repeated until 6 dilutions had been effected. Obviously, 1 cc.
quantities of each of the 6 successive dilutions contain re-
spectively iQ, looj 1,000? io^oooj 100,000) ^lld 1,000,000 cc. of saliva.
One plate each from dilutions 4, 5, and 6 was made, 1 cc. of
the respective dilutions being introduced into lOcc. of nutrient
-{- 1 agar. After thorough mixing, the plates were incubated
aerobically for 24 hours at 37°C. The plate made from dilu-
tion 5 produced 20 colonies, whereas the one from dilution 6
showed no growth. From each of the 20 colonies a cover-glass
preparation stained with gentian violet was made. Microscopic
examination revealed the fact that each of the 20 colonies
was composed of micro-organisms of the coccus type. Transfers
were then made to agar slopes which were incubated at 37 °C.,
for 24 hours. The cultures obtained in this manner were num-
bered from 1 to 20 and kept at 20°C., as stock cultures.
In examining the open air, sterile agar plates were exposed
as indicated in table i.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
53
TABLE I
DATA ON THE COLLECTION OF AIR COCCI
1
Place of exposure.
1
1
Time
of
expo-
sure
Total colo-
nies on plate
after 24 hrs.
at 37°C.
Coccus colo-
nies on plate
after 24 hrs.
at 37°C.
Number
given
to
stock
culture
Remarks
Window sill outside of lab-
oratory, 2nd floor.
15 min-
utes.
14
7
1
21 to 27
incl.
i
1
On slielf, center of labora-
tory room.
do.
i
2
One person in
room. Abun-
dance of Mo-
nilia present.
On table, in reading room.
do.
2
do.
On table, in plating room of
laboratory.
do.
1
Monilia sup-
pressed growth.
On table, in basement
do.
3
On lawn, in garden
do.
9
4
28 to 31
incl.
In living room.
4
do.
7
1
6
1
32 to 37
incl .
Window sill, 4th floor,
downtown section
do.
18
■
Much soot on
plate.
On table, in draughting
room.
do.
1
1
38
One person
in room.
After exposure the plates were covered and incubated aerobically
for 24 hours at 37 °C. Stained preparations of all colonies de-
veloping were made and examined under the microscope. The
coccus forms were transferred to agar slopes, and after incuba-
tion for 24 hours at 37°C., were kept in stock at 20''C. Sev-
eral of the plates exposed in various parts of the laboratory
building were rendered worthless by an abundant growth of
Monilia sitophila.
The method of examining the skin for organisms closely
allied to those characteristic of saliva, was as follows: Test-
54 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
tubes, each containing 10 cc. of distilled water and a piece of
linen 2 inches square, were sterilized in the autoclav for 15
minutes at 15 pounds pressure. Samples were taken from
three parts of the body of a normal individual, namely, the calf
of the leg, the thigh, and the chest. This was accomplished by
briskly rubbing the portion of the body from which the sample
was to be taken with the piece of linen held in sterilized forceps,
and later replacing it in the tube of steriUzed water. From
these dilutions, after being thoroughly shaken, about | cc.
quantities were plated in 10 cc. of nutrient agar. From each
plate 2 coccus colonies were selected from which transfers
were made to agar slopes. These, after 24 hours at 37°C.,
were kept as stock cultures at 20 °C.
There were now in stock a total of 44 pure cultures of Cocca-
ceoe, 20 from saliva, 18 from the open air, and 6 from the skin.
MOKPHOLOGICAL CHARACTERS
The form of the individual cell is of little value in differen-
tiating the species of Cocca^ece, for under conditions favorable
to their growth, all appear as regular spheres. Irregular oval
cells occur at times, but the form usually becomes normal after
cultivation. Some writers lay considerable stress on the value
of cell grouping in the Coccacew as a means of differentiation.
With the utmost care in cultivation and staining, however,
this could not be verified in the cultures under observation.
All the cultures examined contained cells occurring singly, in
pairs, in short chains, and in masses, but in no case did the
cells of any specific culture exhibit a distinct tendency to occur
in any one form. A stained cover-glass preparation showed
various cell groupings in different parts of the same microscopic
field.
Cell grouping was studied in the following manner: An oese
of sterile +1 bouillon was placed on a sterile cover glass, inoc-
ulated with a 24-hour culture of the organism to be examined,
and inverted on a Van Tieghem cell containing a few drops of
sterile distilled water. After sealing the cover glass on the
cell with vaseline, the preparation was incubated for 24 hours
at 37°C. At the end of this time the cover glass was removed
and the drop of water containing the organism allowed to
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLTTION 55
evaporate. Then, 3 drops of mercuric chloride solution were
applied and after 2 minutes washed ofif with distilled water.
Following this, the preparation was treated with a few drops
of 1 per cent acetic acid for 5 minutes, again washed in water,
and finally stained for about 15 seconds with a few drops of
gentian violet. After washing, and drying in the incubator at
37° C, the vaseline was removed from the cover glass with
xylene, and the preparation mounted in balsam and examined
under the microscope. The relation to Gram stain was observed
on 2 and 4-day agar cultures incubated at 20°C. The prepa-
rations were treated with aniline oil-gentian violet for 1§
minutes, with Gram's iodine solution for 1^ minutes, and
finally with 95 per cent alcohol for 3 minutes. The reactions
are recorded as " " (decolorized in both tests), " +
(II J >
a I t J)
(stained in one test and decolorized in the other), and "+ +
(stained in both tests).
CULTURAL CHAEACTERISTICS
1
All cultural characteristics were observed in streak cultures
on agar slants after 14 days' incubation at 20°C., and 37°C.
Such differences as developed between the cultures were
almost entirely variations in color and vigor of surface growth.
Under the latter, 5 types were distinguished as follows:
1. Growth very faint and veil-like, or forming scattered
translucent colonies,
2. Growth better, but still meager.
3. Growth good, but not abundant.
4. Growth abundant.
5. Growth very heavy.
In the study of chromogenesis, apparent differences in pigment
production, due to unequal vigor of growth or evaporation, were,
so far as possible, eliminated. This was accomplished by exam-
ining in each case the same amount of material — a loopfuU —
spread evenly on white drawing paper having a rough surface.
After drying at room temperature, the color of the pigment
produced was compared with the colors as given by Ridgway^
» Color standards and nomenclature. 1912. [Published by the author, Washing-
ton, D. C]
[Vol. 1
56 ANNALS OF THE MISSOURI BOTANICAL GARDEN
This author uses as a basis the solar spectrum with its six
fundamental colors and intermediate hues, augmented by a
series between violet and red not in the spectrum.
BIOCHEMICAL REACTIONS
The production of indol was investigated in 5-day peptone
broth cultures incubated at 37°C. One cc. of a 10 per cent
sulphuric acid solution was thoroughly mixed with the broth
■
culture, and then 1 cc. of a freshly prepared 0.01 per cent
solution of sodium nitrite was carefully run in on top of the
mixture. The appearance of a pink ring at the juncture of
the nitrite solution with the acid-peptone solution, was regarded
as an indication of the presence of indol. A blank determina-
tion for purposes of comparison was made in each case. The
action on neutral red broth as regards change in color was
observed in cultures incubated for 5 days at 37°C., in the
presence of hydrogen.
The organisms were further grown in solutions of nitrate
broth to determine whether or not reduction takes place, and
if so, whether to nitrite or to ammonia. In carrying out the
test a tube of nitrate broth was inoculated with the organism
to be tested, and incubated for 4 days at 37°C., an uninoculated
tube of nitrate broth being similarly treated to serve as a check.
At the end of 4 days, 3 cc. of the broth were removed to a clean
test-tube, and 2 cc. each of a naphthylamine solution and of a
sulphanilic acid solution added. The development of a red
color indicates the presence of nitrites, the intensity of the
color being proportional to" the amount of nitrites present in
solution. To test for ammonia in the remaining portion of
the culture, a few drops of Nessler's solution were added.
The appearance of a yellow color or precipitate indicates the
presence of ammonia. In studying the liquefaction of gelatin
by the cocci under observation, the extent of the action
only was determined. This was accomplished by spreading
a suspension of the organism over the surface of gelatin in
10 mm. tubes. It was found that the amount of material used
in this inoculation did not affect the total amount of liquefaction,
i.e., whether the amount of transferred material was large or
1914]
• NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 57
small the extent of the liquefaction after 30 days' growth at
20°C., was the same with any one organism.
In the study of the action on sterile certified milk particular
attention was paid to the coagulation of the milk and to the
production of acid. Observations were further made on the
effect of the organisms on lactose, saccharose, mannite, salicin,
inulin, sorbite, raffinose, and rhamnose. The medium in which
these organic substances were used was prepared according
Dr. Houston's formula, as follows
Liebig's beef extract
1.0 per cent
1 .0 per cent
Peptone
Organic compound to be tested 1.0 per cent
Sodium bicarbonate 0.1 per cent
10 per cent litmus solution 1 .0 per cent
The medium, neutral in reaction to litmus, was sterilized for
15 minutes at 15 pounds pressure in 500 cc. containers, from
which sterile fermentation tubes, provided with glass caps,
were directly filled. In doing this it was necessary to take
utmost precautions to obviate any possibility of contamina-
tion. The various organic media thus prepared were inocu-
lated, not from an agar slope, but from a 48-hour broth culture.
Gas formation and the production of acid in the several media
were observed after 3 days' incubation at 37 °C.
DISCUSSION OF RESULTS
A thorough study of the results will now be made with a
view of finding, if possible, some characteristic or group of
characteristics, morphological or biochemical, which may be
used in differentiating the salivary cocci from the coccus forms
of the air and the skin.
The cell grouping varies throughout, there being
ment characteristic of any particular group.
As observed
the forms occur in groups, chains, and pairs. As regards the
deportment of the various organisms toward the Gram stain,
it was noted that all of the salivary cocci gave a positive reac-
tion in both tests; of those from the air, 3 were positive in
the two tests, 8 alternately negative and positive, and 6 negative
throughout; of the skin cocci, 4 were positive and 2 negative
*
[Vol. 1
58 ANNALS OF THE MISSOURI BOTANICAL GARDEN
in both tests. This stain, as may readily be seen, is of no
differential value here, for, although the salivary cocci react
positively throughout, both positive and negative reactions
occur among the air and skin forms.
The production of indol among coccus forms is very uncom-
mon. Of the salivary cocci under observation, none produced
indol, and of the air and skin forms only one from each group
produced it. The change of color in neutral red broth is,
apparently, more frequently brought about by the salivary
cocci than by the air and skin forms, but this difference is not
sufficiently well marked to be of differential value. Of the
20 salivary cocci, 12 produced fluorescence, whereas only 1 of
the air and none of the dermal forms produced this change.
All of the forms under observation reduced nitrates to ammonia.
Of the salivary forms, 14 out of 20; of the air cocci, 5 out of 18;
and of the skin cocci, 5 out of 6, reduced nitrates to nitrites.
^ +
It thus appears that the reduction of nitrates to ammonia
is very common among members of the Coccacece, but that the
reduction to nitrites only is variable and not characteristic
of any one type.
The average amounts of gelatin liquefied after 30 days'
growth at 20°C., are as follows: by the salivary cocci, 2.8 cc;
by the air forms, 1.9 cc; and by those of the skin, 1.4 cc. Fif-
teen out of 20 of the salivary organisms, 15 out of 18 of the air
forms, and 4 out of 6 of the skin cocci, liquefied gelatin. Sum-
ming up the results obtained from the experiments on gelatin
liquefaction, it is to be. noted that, in general, the salivary
cocci liquefy gelatin more readily than do the air or skin forms,
but aside from this it is apparent that there is nothing to warrant
the use of gelatin as a differential medium.
The results of the experiments on vigor of surface growth
on agar slopes at 20°C., and 37°C., are given in table ii. While
it may be said, in general, — from the results given in this
table — that the salivary cocci grow somewhat more vigor-
ously at 37°C. than at 20°C., the air forms better at 20°C.
than at 37°C., and the skin organisms about equally well at
the two temperatures, the differences are not sufficiently pro-
nounced to impart to the factor of vigor of surface growth
any marked value as a differential characteristic.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
59
TABLE II
DATA ON THE VIGOR OF SURFACE GROWTH OF AIR. SALIVARY. AND DERMAL COCCI
Source of
organism
Temperature
of incubation
No. of
cultures
used
Growth characteristics
■
Saliva
Air
Skm
Saliva
Air
Skin
20°C.
20''C.
20°C.
37°C.
37°C.
37°C.
20
18
6
20
18
6
No
growth
1
Very
faint
Mea-
ger
Good
Abun-
dant
i*
3
1
2
1
1
7
2
7
9
10
4
2
13
6
4
7
1
Very
heavy
6
1
In the following enumeration are listed the colors of the
various pigments produced by the air, skin, and salivary cocci,
the figure on the left having reference to the number in Ridgway
corresponding to the particular pigment produced:
Salivary cocci
15" d Light pinkish cinnamon
3
15" c Intermediate between light pinkish and
pinkish cinnamon
15" b Pinkish cinnamon
15" a Intermediate between pinkish cinnamon
4
4
15"
21' e
and cinnamon
Cinnamon
Intermediate between massicot and
yellow
3
1
23' f Naphthale
19 f Maize yell
19' b Mustard y
1
1
1
1
15
One gave too little growth for determination of
the color.
60 ANNALS OF THE MISSOURI BOTANICAL GARDEN
ITOL. 1
Air cocci
21' i Massicot yellow
21' e Intermediate between massicot and straw
yellow
21' d Straw yellow
21' b Amber yellow
19' d Naples yellow
19' b Mustard yellow
19' Primuline yellow
19 f Maize yellow
19 d Buff yellow
3' b Light Jasper red
One form did not grow.
4
2
1
1
1
4
1
1
1
1
\ 16
Skin cocci
19 f Maize yellow
19 d Buff yellow
19 b Apricot yellow
21' e Intermediate between massicot and
yellow
White
1
1
2
5
1
1
J
At first glance the color of the pigments produced by the
organisms would seem to furnish one mode of differentiation.
In the majority of cases the salivary cocci produced cinnamon
colored pigments, whereas pigments of a yellow color were
usually produced by the air and skin forms. Closer inspection
shows, however, that some of the salivary cocci, as well as
the air forms, produce a maize yellow and a mustard yellow
pigment; also that a maize yellow pigment and one inter-
mediate between massicot and straw yellow are produced
by representatives of both the salivary and skin cocci. It
is apparent that these intergradations make the factor of pig-
ment production largely inapplicable as a differential test.
In milk the salivary cocci with one exception produced acid
and coagulated the medium, whereas none of the air forms
and but one of the skin cocci gave this combined reaction.
This attaches to milk considerable value as a differential me-
dium. In the media containing the various organic substances
1914]
NOLTE — SALIVARY ORGANISMS AND AIR TOLLUTION 61
sugars, etc., none of the coccus forms produced gas. All but
one of the salivary cocci produced acid in the lactose medium,
whereas none of the air cocci and but one of the skin forms
deported themselves in this manner. This marks lactose broth
as another medium of differential value.
, The salivary cocci with but one exception produced acid
in saccharose, the single exception being the organism which
produced no acid in the lactose medium. Two air cocci and
one skin form also produced acid in saccharose, but notwith-
standing these exceptions, it appears that saccharose is a third
valuable differential medium. In the mannite, salicin, inulin,
sorbite, rafhnose, and rhamnose broths none of the organisms
produced acid, thus marking these organic substances as of
no value in differentiating the types of cocci under investigation.
SUMMARY
In reviewing the preceding discussion of results we find
three media, namely, lactose and saccharose broths, and milk,
which are of value in differentiating the cocci most character-
istic of saliva from those of the air and the skin. One of the
salivary coccus forms did not produce acid in lactose and
saccharose broths and formed neither acid nor clot in milk.
This may have been, and probably was, an air or skin form.
Among the air cocci are two which vary somewhat from the
remaining air and skin forms in that they produce acid in
saccharose broth. Neither of them, however, produces acid
in lactose broth, nor acid or clot in milk and in these respects
they differ markedly from the characteristic salivary forms.
Of the skin cocci one gave the characteristic reactions of the
salivary organisms, and it is not at all unlikely that this was a
salivary coccus. In general, then, it appears that the organism
most characteristic of saliva is a coccus form which produces
acid in lactose and in saccharose broths, and acid and clot in
milk.
FURTHER TESTS
To further test the validity of the reactions above referred
to as furnishing a reliable means of differentiating between
salivary cocci and those of other origin, two additional samples
of saliva, from two different individuals, were examined, —
[Vol. 1
62
ANNALS OF THE MISSOURI BOTANICAL GARDEN
one from a middle aged white person (A), the other from a
colored person (B).
The samples were collected and treated in a manner similar
to that outlined in the early part of this paper. In the first
case (A), transfers were made from all colonies on two plates,
representing a dilution of one part saliva in ten billion.
These subcultures, all of cocci, were numbered from 1 to 17
inclusive.
In the second case (B), transfers were made from 36 colonies
which developed on one-third of a plate representing a dilution
of one part saliva in ten billion. The entire series of cultures,
numbered from 1 to 36 inclusive, although made from 36
colonies from a plate containing a total of 100 colonies, were
found to be made up of coccus organisms. After being incu-
bated in + 1 nutrient broth for 2 days at 37°C., each of the
■
cultures from samples (A) and (B) was transferred to the three
differential media, — ^lactose and saccharose broths, and milk.
The results recorded in tables iii and iv were observed after
3 days' incubation at 37°C. No gas was produced in any of
the sugar media. A blank determination gave negative re-
sults throughout on the three media.
TABLE III
■
REACTIONS
OF SALIVARY
COCCI (A)
1
ture
JS
t
Milk
p
"o
1^
:3
o
Milk
+3
1
O
Milk
•
9
QD
m
3
u
QD
Li
1
1
1
o
o
M
1
"o
O
M
"o
I o
u
Acid
Qot
o
1
7
<J
Acid Clot
#
o
Acid Clot
o
05
+
+
+
+
■
03
+
+
+
1
+
+
-— ■ ^ ^^^^^^*
1
+
1
1
+ ^
+
■
+
+
+
o3
+
+
+
+
+
+
c3
1
+ ;
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
1
+
03
+
+
+
+
+
1
+
+
+
1
+
+
1
1
13
14
+
2
8
+
3
9
15
+
4
10
16
+
5
+ +
1
11
17
+
6
+
+
12
+indicateB positive reaction.
O indicates negative reaction.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
63
From the above table it is evident that all but one of the
coccus forms in series (A) produced acid in lactose and saccha-
rose broths, and acid and consequent clotting in milk. The
one exception was probably an air coccus.
TABLE IV
REACTIONS OF SALIVARY COCCI (B)
culture
2
Xi
+
1
+
1
+
+
+
+
+
+
+
+
+
1
1
+
+
' +
1
1
1
+
1
+
+
+
+
1
Milk
1
w No. of culture
o
s
+
+
+
+
+
+
+
+
+
o
1-,
1
+
+
1
+
+
+
+
+
i
+
+
+
+
+
Milk
3
o
25
26
27
28
1
29
o
Xi
OP
QQ
o
o
+
+
1
1
+
1
1
1
1
+
+
+
^ +
f
i
+
+
+
+
x:
o
x^
xi
o
03
1
i
+
1
+
+
+
+ ^
Milk
o
1
Acid
+
+
+
+ J
+
+
+
1
+
+
+
1
Clot
+
+
+
+
+
-■
+
+
+
+
+
Acid Clot '
+ +
Acid
+
+
+
1
1
+
1
1
1
Clot
2
1
1
14
15
16
+ +
+
1
3
4
+
+
T
+
+
i
+
+
+
+
■
+
+
■
+
+
+
+
1
+
+
5
17
18
+ ,+
6
30
31
+
+
7
19
+ +
+
8
20
32
+
1
+
+
+
+
+
+
+
+
+
1
+
9
21
33
+
10
22
34
+
11
23
35
+
X2
24
36
+
+iiidicate8 positive reaction.
O indicates negative reaction.
In series (B), 32 out of the 36 cocci reacted positively through-
out on the three differential media. The remainder were positive
with saccharose, but negative with lactose and milk, agreeing
in this respect with the two air cocci to which reference has
been made.
The reactions of the organisms from saliva (A) and (B)
6
[Vol. 1
64 ANNALS OF THE MISSOURI BOTANICAL GARDEN
further indicate that the production of acid in lactose and sac-
charose broths, and a similar production, together with clot, in
milk, are characteristic reactions of the salivary cocci.
CONCLUSIONS
From the results of the preceding experiments it appears
that a method applicable for the detection of the organisms
characteristic of human saliva has been developed.
It must be acknowledged that the number of organisms
examined is comparatively small, especially where those of
the air and the skin are concerned. An absolute test of the
validity of the adopted mode of identification would necessitate
the examination of many hundreds of strains of cocci from
numerous sources.
Nevertheless, the characteristic reactions of the salivary
cocci examined seem to be sufficiently definite to warrant the
assumption that the most characteristic salivary organism is
a coccus form which produces acid in lactose and saccharose
broths, and acid and clot in milk.
The Relation of the Most Characteristic Salivary
Organism to the Pollution of Air
Having identified the most characteristic salivary organism,
the next problem is to isolate it from the air. Its frequency
of occurrence must also be determined, as this often serves as
an index to the degree of pollution. The isolation of the organ-
ism and the determination of its frequency of occurrence can be
accomplished simultaneously.
Then come the 'problems (1) of devising an air-collecting
apparatus suitable for all occasions, and (2) of determining the
quantity of air to be examined and the terms by which the
sanitary quality of the air shall be expressed.
In searching for a means of expressing the sanitary quality of
air, let us consider the manner in which this is accomplished
in drinking water. Authorities differ markedly on this subject.
Shall a water be considered safe or unsafe for drinking purposes
if B. coll is present in a 100 cc. sample, or shall its presence or
absence in 10 cc. or 1 cc. samples be taken as the basis for the
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
65
classification? In lieu of a definite standard let us assume the
following table 1;
TABLE V
PRESUMPTIVE TEST FOR B. COLI IN WATER
Sanitary quality
cc.
0.01
Safe
Reasonably safe
Questionable
O
cc.
0.1
cc.
1.0
O
cc.
10.0
cc.
100
O
Probably unsafe
+
+
+
o
o
+
+
+
+
Unsafe
+
+
+
+
+
+
+
+
+ indicates positive presumptive test for B. coli.
We shall now endeavor to prepare a similar table for the purity
of air, expressed in the number of salivary cocci present in given
volumes. In the normal life processes, the volume of air inhaled
is obviously much greater than the volume of water consumed,
and this fact must be taken into consideration in establishing
for the bacteriological examination of
It has
been estimated that the tidal air, i.e., the air taken in with each
inspiration and given out with each
normal adult when at rest, to one
amounts
one-half liter. Assuming the
average frequency of respiration to be 15 per minute, the amount
of air inhaled in one minute is 7^ liters, in one hour, 450 liters,
and in one day, at least 10,000 liters. Taking the average
amount of unboiled water drunk in a day as 2 liters, it would
appear that 5,000 times as much air as water is required daily.
Hence, the following table, based on table v, may be used ta
express the sanitarv aualitv of air:
» Whipple, G. C. On the practical value of presumptive tests for B. coli in
water. Techn. Quart. i6:18 e. ra. 31. 1903.
66 ANNALS OF THE MISSOURI BOTANICAL GARDEN
TABLE VI
TEST FOR CHARACTERISTIC SALIVARY COCCI IN AIR
[Vol. 1
Sanitary quality
cc.
50
cc.
500
1
cc.
1
5,000
cc.
50,000
1
cc.
500,000
1
Safe
+
Reasonably safe
1
1
+
+
Questionable
1
1
i
+
+
+
Probably unsafe
+
+
+
1
+
+
Unsafe
+
+
+
+
+ indicates positive reaction in the three differential media adopted.
APPARATUS AND TECHNIQUE
■
As it was the intention to collect samples of air in places other
than the laboratory, a portable apparatus was necessary. As
devised, it consists essentially of a sand filter, a support for same
with an attachment for alternately opening and closing the
exhaust and
and a bulb, having a capacity of 16
with the required amount of rubber tubing
The sand filter is of the standard type.
plate 2.)
tube
It consists of
mm. lone and 10 mm. in diameter, fitted with
hole rubber stopper, through which passes a piece of 6 mm
tubing
This stopper, with its tubing, forms the support for a
disc of bolting cloth with a 10 mm. layer of very fine
sand that passes through a 100, and is retained
mesh
d 12
The support consists of a rectangular piece of wo(
X 1 X I inches, fitted with a double pinch cock arrangement.
Clamps for holding the filter in position are also provided. The
rubber bulb is connected to the apparatus in such a manner that
when pressure is applied to the former and the pinch cock opened,
the air contained in the bulb is expelled through the exhaust
any way
Up
without disturbing the sand in the filter in
operation occupies but a few seconds of time,
the pinch cock, and immediately thereafter the bulb, the
drawn through the sand.
This
releasing
1914]
NOLTE — SALIVARY ORGANISMS AND AlR POLLUTION 67
The volume of air exhausted from the bulb at each pressure
was determined as follows: The bulb, filled with water, was
weighed. Pressure was then applied, forcing out the water,
after which the bulb was again weighed. The difference in
weight in grams is approximately the volume of air in cc. ex-
hausted by a similar pressure. In the calibrations the results
varied but slightly. By placing the fingers on the bulb in a
certain fixed position each time, it was found that the bulb could
be made to deliver 300 cc. of air at each exhaustion and, conse-
quently, to receive 300 cc. of air at each release of pressure.
It was, of course, necessary to have all joints air-tight, this being
accomplished by making all connections with rubber tubing and
glass and using plenty of overlap.
The sand filter, after being plugged at both ends with cotton,
was sterihzed for 30 minutes at 15 pounds pressure. The rubber
stopper support was allowed to fit very loosely into the tube dur-
ing sterilization in order to prevent setting of the rubber. After
the apparatus was removed from the autoclav, the stopper was
immediately fitted in tightly, thus rendering the connection air-
tight. The sand filter was always used within 24 hours after
sterilization. It was connected to the support as shown in
plate 2.
When operated in public buildings or conveyances, the sup-
port, with the filter, was wrapped in stiff paper in such a manner
as to permit of the easy operation of the pinch cock and exhaust.
The apparatus thus wrapped was held in the left hand and from
it heavy rubber tubing passed down the left coat sleeve and then
diagonally across to the right coat pocket where it was connected
to the bulb. This rendered the whole apparatus inconspicuous.
The bulb was operated with the right hand, the pinch cock with
the left. A test tube with a sterile cotton plug was always
carried, the latter being used to replace the plug which was
removed from the intake of the filter at the beginning of the
experiment.
The plating was always carried out within 30 minutes after
the sample was obtained. The sand from the filter was carefully
poured into a 100 cc. flask containing 15 cc. of sterile distilled
water. The bolting cloth, which had a tendency to stick to the
rubber stopper, was removed with sterile forceps and introduced
[Vol. 1
68 ANNALS OF THE MISSOURI BOTANICAL GARDEN
into the flask. The contents of the flask were thoroughly shaken
and aliquot portions, as shown in table ix, were plated with 10
cc. of +1 nutrient agar. In plating, the introduction of much
sand was avoided in the following manner : The end of the pipette
was held immediately above the bottom of the flask while the
: liquid was being drawn up to a point slightly above the gradua-
': tion mark. After a few seconds, enough of the sandy liquid was
allowed to run back into the flask to leave the water just at the
mark. During this short interim a large proportion of the sand
settled in the tip of the pipette and was returned to the flask
as the liquid was lowered to the mark. Blanks were plated
several times during the course of the experiments, but no
growth developed in any case.
The plates were in aU cases incubated for 4 days at 37°C.,
after which the number of bacterial colonies present in each was
determined. Finally, all, or a representative number, of the
colonies were examined for the presence of coccus forms,
table IX.)
The coccus colonies developing on agar are, as a rule, very
small and often grow in the deeper strata of the medium. This
renders the transfer diflacult especially when two are to be
made from the same colony — one for the stained preparation
and one for the agar slope to be used as a stock culture. The
difficulty was partiafly obviated by subculturing (from all the
colonies in certain selected plates) to agar slopes, and incubating
the latter at 37 ''C. After several days an examination served
to eliminate the bacilli and moulds, leaving only the coccus
cultures which were later examined for the presence of the sali-
vary forms. In this examination the three differential media
described above were used.
SOURCES OF SAMPLES
As the investigation in hand seeks to discover a relation be-
reen the presence of a characteristic salivary organism and the
►llution of air, it was thought best to collect the samples of
r under normal conditions, i.e., conditions which are met with
every-day life.
Public conveyances, on account of their usually crowded condi-
»n and frequently inefficient ventilation, suggested themselves
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
69
favorable
for tests. Hence, a local street car was
chosen as a source for air sam
The often poorly ventilated
but well filled motion picture theatres furnished another
posedly promising sam
The third locality chosen
for
samples was a local 5 and 10
It
was thought that this would furnish an ideal source of contami
nated ak because of the large crowds of people who are con
tinually voicing theu- sentiments and desires. In order to deter
mine whether or not the salivary
organism
mosphere which is not in immediate contact with human being
and which is open to the ventilation of nature,the fourth samp!
taken from the
DISCUSSION OF THE EXPERIMENTS
The experiments in table
dates on which the tests w
are arranged a
made. But for
rding
the
this discussion the experiments will be taken up according to
the source of the samples.
Experiment i.— This experiment was carried out primarily
to test the apparatus. The air sample was taken in a labora-
tory on the second floor of an old building. There were usually
at least two people present in the room, and practically no
ventilation was provided, the doors and windows being con-
stantly closed. The apparatus used differed from that used in
the remaining experiments in that two sand filters were used
in tandem instead of the usual one. During the 15 minutes of
operation, 7,800 cc. of air were drawn through the sand of both
filters at the rate of 520 cc. per minute.
The sand of the first filter was introduced into 15 cc, that of
the second into 6 cc. of sterile distilled water. Quantities of
both solutions were plated with the following results :
TABLE VII
[Vol. 1
70 ANNALS OF THE MISSOURI BOTANICAL GARDEN
The reactions of the cocci isolated from the air in the fh-st
filter showed that there was one salivary coccus form present.
The remaining three gave negative reactions on the three differ-
ential media. It should be noted that out of the 15 cc. of solu-
tion from the first filter, only 4 cc. were plated. Eight organ-
isms were present in the quantity examined, making a total of 30
in the entire solution. One characteristic salivary coccus form
developed in the portion examined; making, according to the
law of averages, a total of 4 in the entire solution. The total
volume of air examined being 7,800 cc, the frequency of occur-
rence of the salivary coccus is 1 in 1,950. According to table
VI, the sanitary quality of the air of the room was "probably
unsafe" at the particular time at which the sample was taken.
EXPERIMENTS 3, 5. 6, 8, 10
These experiments were carried out in a local street car.
The same car line was chosen for all of the experiments in order
to eliminate as many variables as possible, such as construction
of car, capacity, rate of locomotion, etc. The car was of the
ordinary "pay-as-you-enter" type now in use in St. Louis.
It had a seating capacity of about 44 people, and could accom-
modate approximately 40 more standing indoors. The air
space in the car in question was about 2,500 cubic feet, or ap-
proximately 30 cubic feet for each passenger when the car was
filled to its capacity.
As the samples were taken at a time when the outside tem-
perature would not permit the windows to be open, the question
of ventilation was carefully studied. As is usually the case,
the transoms were tightly closed, and only when the front and
rear doors of the car were open at the same time was there an
opportunity for a complete renewal of the air. This never
happens when the car is in motion, and there is probably never
a complete renewal of air unless a strong wind is blowing, thus
causing a draught when the car is at a standstill, with both doors
open. This particular car was provided with four vents in the
roof which could be opened or closed at will. In several of the
experiments some of the vents were open; in others, all were
closed.
The degree of pollution of the atmosphere in such a car de-
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 71
pends, of coursej on the amount of coughing, sneezing, speaking,
etc., of its occupants. A car may be very crowded but if no
coughing, etc., is going on, there will, theoretically, be no pollu-
tion of the atmosphere from saliva. Again, if there is much
talking, etc., among those present, the atmosphere may be
greatly polluted by the dissemination of particles of saliva from
the mouth.
The samples were always taken in the early morning between
the hours of six and seven, when the majority of the laboring
class are on their way to work. The tendency of the passengers
at this time of the day is to be quiet, as the morning paper is of
absorbing interest to a majority. The samples of air were taken
in the center of the car, the opening of the apparatus being about
4 feet from the floor level. In these experiments the apparatus
described above was used. In all cases 10,800 cc. of air were
drawn through the sand filter at the rate of 900 cc. per minute.
The sand was introduced into 15 cc. of sterile distilled water
and plated as shown in table ix.
The experiments carried out in street cars will now be taken
up in order and the results discussed. If it can be shown that
the characteristic salivary organism is present in the air of these
cars in sufficient quantity, and if it can later be proved that this
salivary organism is not present in the open air, it follows that
the atmosphere in these cars is being polluted by the dissemina-
tion of particles of saliva from the mouth.
Experiment 3. — While the air sample was being taken for this
experiment, 44 people were seated in the car, but none were
standing. Out of the 20 colonies appearing on the plate (see
table ix), 9 were of bacilli and 11 of coccus forms. Inoculated
into the 3 differential media, 8 of the latter reacted negatively
in all three media, 1 negatively on lactose and milk, but posi-
tively on saccharose, and 2 gave positive reactions in all three
media.
It will be recalled that mention has been made of several
organisms, both among the salivary and air cocci, which gave a
positive reaction with saccharose, but reacted negatively with
lactose and milk. The one above referred to as reacting in
this manner is probably one of these unidentified coccus
forms which seem to be present in both saliva and air. Out of
[Vol. 1
72 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the 11 cocci present, therefore, two were of the characteristic
salivary type, and as only one-third of the sample was plated,
a total of 6 may have been present in the entire volume of air
examined, or a frequency of occurrence of 1 in 1,800. Accord-
ing to table VI, the sanitary quality of the air was ''probably
unsafe."
Experiment 5. — During the sampling process for this experi-
ment, 44 persons were seated and approximately 30 standing.
Of the 26 colonies which developed on plate 5 (see table ix),
14 were of bacilli, 2 of streptothrix, and 10 of cocci. When
transferred to the three differential media, all of the latter gave
negative reactions, indicating that the air in the car at the time
of this experiment was "safe."
Experiment 6. — At the time of sampling, 44 persons were
seated and 30 were standing. On account of the large number
of colonies present, only a representative sector of plate 1 — com-
prising one-twelfth of the total area— was examined (see table ix).
On this area 21 colonies were counted, 5 bacillus and 16 coccus.
On the three differential media, 4 of the latter gave nega-
tive reactions throughout, 6 were negative on lactose and milk
but positive on saccharose, and 6 gave positive reactions on all
three media. It follows that 6 salivary cocci were isolated from
one-twelfth of the plate, making a total of 72 from the entire
plate, or of 1,080 from the total volume of sand solution, — a
frequency of occurrence of 1 in 10. According to table vi, the
air in the car at the time of the experiment was ''unsafe."
Experiment 8. — The number of persons seated and standing
was the same as in experiment 6. On the plate examined (see
table ix), 34 colonies developed — 15 bacillus and 19 coccus. On
the three differential media the coccus forms reacted as follows:
Twelve gave negative reactions throughout, 6 were negative
on lactose and milk but positive on saccharose, and 1 was nega-
tive on lactose and saccharose but positive on milk. The
last form was found, after again staining with gentian violet
and examining under the microscope, to be a short bacillus. It
is to be noted that 6 organisms of the unidentified coccus type
were again present. No characteristic salivary cocci were
present, thereby marking the air of this particular car as "safe"
at the time of the experiment.
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 73
Experiment 10. — During this experiment, 44 persons were
seated and 15 standing. It was noted that one transom was
open. The plate examined (see table ix) gave a total of 12
colonies, of which 5 were of baciUi and 7 of cocci. Of the latter, 6
reacted negatively on all three of the differential media, whereas
1 gave a positive reaction throughout. This makes the fre-
quency of occurrence of the characteristic salivary coccus form
1 in 3,600, and, according to table vi, marks the air in this car
as "questionable" at the time of the experiment.
Summarizing the car experiments, it is to be noted that in
three out of five cases the characteristic salivary coccus form
was isolated, and in such quantity as to mark the air of one
''unsafe," that of another ''probably unsafe," and of a third
"questionable."
EXPERIMENTS 9 AND 11
These experiments were carried out in a local vaudeville house.
The construction of the building appeared modern in every
respect. The lower floor had a seating capacity of about 2j000,
while the balcony accommodated approximately 1,000 people.
The house was filled with spectators on the occasions when the
samples were taken. Upon inquiry, after the surprisingly good
results given below were obtained, it was found that the build-
ing was well ventilated by one of the modern appliances for
this purpose, whereby the volume of air in the building (about
90,000 cubic feet) was being renewed to a greater or less extent
every seven-tenths of a minute. For the collection of the air
samples, the same apparatus was used as in the street car
experiments, 10,800 cc. of air being drawn through the sand filter
at the rate of 900 cc. per minute. The sand was introduced into
15 cc. of sterile distilled water and platings were made as indi-
cated in table ix.
Experiment 9, — The air sample was obtained near the center
of the lower floor of the building about 60 feet from the stage.
The entire lower floor was packed, and in addition about 100
or more persons were standing in the rear. On the plate ex-
amined; a total of 14 colonies developed, — 3 mold, 9 bacillus
and 2 coccus. Molds were very abundant on the other plates.
One of the coccus forms reacted negatively on lactose and milk
but positively on saccharose, whereas the other gave negative
74 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
reactions on all three differential media. The presence of the
single unidentified coccus is again noted. No salivary coccus
forms were isolated, from which fact it appears that the air in
the particular location from which the sample was taken was
"safe."
Experiment 11. -~Thm sample was taken on the balcony of
the building, about 10 feet from the rear wall. Every seat was
occupied. As indicated in table ix, two plates were examined.
On the first, 7 colonies developed— 3 streptothrix, 3 bacillus, and
1 coccus. On the second plate 3 colonies appeared, all of which
were of bacilli. The reaction of the coccus was negative on the
three differential media, thereby indicating that the sample of air
taken was free from salivary coccus forms and therefore "safe."
In summing up the results of the experiments carried out in
the vaudeville house it is to be noted that in both cases no sali-
vary coccus forms were found. Table ix further shows that
the total number of organisms found per unit volume of air
was smaller than in the street car experiments.
EXPERIMENTS 4 AND 7
These samples were obtained in the basement of a local 5 and
10 cent store. The ceiling was rather low, being only about
9 feet from the floor level, the entire basement having a volume
of about 72,000 cubic feet. The samples in these experiments
were taken in the midst of a crowd gathered to listen to a singer
advertising songs. Little attention was given to the matter
of ventilation until after the results of the experiments were
obtained. Subsequently, however, investigation revealed the
fact that ample provision had been made for ventilation.
Transoms at the level of the sidewalk provide openings to the
outside; along the inside wall and near the ceiling are revolving
fans about 20 feet apart. These keep the air in circulation
until it is drawn out by a suction fan situated in one corner,
about 2 feet from the ceiline:. The
same sam
was used as in the preceding experiments. As before, a total
of 10,800 cc. of air was drawn through the sand filter in each
sampHng at the rate of 900 cc. per minute. The
plated as shown in table ix.
sam
4- — The air sample for this experiment
1914]
NOLTE — SALIVAKT ORGANISMS AND AIR POLLUTION 75
in the midst of a crowd of about 100 people in front of a counter.
On the plate examined, a total of 36 colonies developed — 16
bacillus and 20 coccus. Of the latter, 18 gave negative reactions
throughout on the three differential media, and 2 reacted nega-
tively on saccharose and milk but positively on lactose, the
latter sugar being fermented. No salivary coccus forms were
isolated, indicating that the air in the basement at the time of
the experiment was "safe."
Experiment 7. — This air sample was taken under practically
the same conditions as in the previous experiment except that
only about 50 people were in the crowd. The plate examined
contained 2 streptothrix, 13 bacillus, and 8 coccus colonies. All
of the cocci gave negative reactions throughout on the three
differential media. No salivary coccus forms were found, which
fact leads again to the conclusion that the sanitary quality of
the air during the experiment was ''safe."
EXPERLMENTS 2, 12, 13, 14
These experiments were performed outdoors. The air sample
for experiment 2 was collected in a railroad switch yard at a time
when there was no traffic. The samples for experiments 12, 13,
and 14 were collected in the immediate vicinity of large storage
basins belonging to the local water works and located 300 or 400
feet from the bank of the Mississippi River. The apparatus
used was the same as that employed in the previous experi-
ments.
Experiment 2. — The outdoor temperature was 29°F., and
samnle was bein
A
of 22,500 cc. of air was drawn through the sand filter at the rate
of 750 cc. per minute — the operation extending over a period of
30 minutes. Samples were plated as shown in table ix. Of
the 3 plates examined, plate 1 yielded 2 bacillus colonies; plate
2, 1 streptothrix, 1 bacillus, and 6 coccus colonies; and plate 4,
2 mold, 1 streptothrix, 1 bacillus, and 2 coccus colonies. All
of the coccus forms were grown on the three differential media,
7 giving negative reactions throughout, while 1 reacted positively
on saccharose and negatively on lactose and milk. The latter
organism is one of the unidentified coccus forms previously
referred to. No characteristic salivary cocci were found, in-
76
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
dicating that the sanitary quality of the air examined was
"safe."
Experiment 12 —
m
i^- — At the time the air sample was being taken,
a slight drizzling rain was falling, accompanied by considerable
wind and a temperature of 45°r. Prior to that time it had been
raining continuously for about 24 hours. A total of 10,800 cc.
of air was drawn through the sand filter at the rate of 900 cc.
per minutcj the apparatus meanwhile being held about 5 feet
above the ground level. The sand of the filter was introduced
into 15 cc. of sterile distilled water, from which Dlatines were
made,
with the
Table viii gives the details of the experiment, togeth
results obtained.
TABLE VIII
Plate
number
Quantity
plated
Total no.
of colonies
1
3
4
1 cc.
1 cc.
5 cc,
5 cc.
No. of bac-
teria and
molds
Coccus
colonies
No. of sali-
vary cocci
9
3
6
2
3
4
7
7
2
Attention should be called to the fact that on plate 1, in which
only 1 cc. of the solution was used, 9 colonies developed— 7
coccus and 2 bacillus—, while on plate 4, in which 5 cc. of the solu-
d, only 6 colonies appeared
and 4 bacillus
Furthermore, the 7 colonies in plate 1 proved to be of salivary
cocci, whereas none of these organisms were present among the
of
These results unquestionably indicatelocal
tamination.' It is difficult to say just where th
took place. Obviously
did not occur during the collection
of the sample or even during the mixing of the sand
for had this been the case all of the plates should have shown
the greater number should have occurred
salivary cocci, and
hich larger quantities of
plated. In all probability plate 1 was locally contaminated
Experiment
While the sample of air was being taken for
this experiment, the temperature was 63 °F., a light bree
bl
and the sky was very cloudy although no rain had
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION 77
fallen during the preceding 18 hours. A total of 10,800 cc. of
air was drawn through the apparatus at the rate of 830 cc. per
minute. Samples of the sand solution were plated as shown in
table IX. It is to be noted that in those plates containing 1 cc.
of the solution no colonies developed, whereas in those contain-
ing 5 cc, 1 bacterial colony appeared in each. Attention is called
to the consistent results in this experiment to emphasize the
fact that the inconsistencies in experiment 12 are due to local
contamination. No salivary cocci were found.
Experiment 14- — The air sample for this experiment was taken
on a bright, clear day, with a rather strong wind blowing and a
temperature of 55°F. A total of 10,800 cc. of air was drawn
through the sand filter at the rate of 1,080 cc. per minute.
Samples of the sand solution were plated as shown in table ix.
Of the 3 coccus forms, 2 gave negative reactions on all three
differential media, whereas 1 was positive on saccharose and
negative on lactose and milk. The latter will be recognized as
one of the unidentified coccus forms. No salivary cocci were
isolated.
Summarizing the open air experiments, it is to be noted that,
barring the locally contaminated plate 1 in experiment 12, the
characteristic salivary coccus form was not isolated; further-
more, that the total number of organisms in the open air is
comparatively low.
SUMMAKY AND CONCLUSTONS
Examining the entire series of experiments it appears that
in the majority of cases where ventilation was obviously inad-
equate, the characterisitic salivary coccus form was isolated.
On the other hand, the form could in no case be found where
am
artificial or natural ventilation existed
It has been shown that the most characteristic salivary
organism can be differentiated and identified; also, that this
characteristic organism can be isolated from the air.
In the experiment carried on in one of the street cars in which
there were many passengers, the characteristic salivary coccus
form was found to be present in such quantities as to indicate
that the air in this car was "unsafe." It was later shown that
[Vol. 1
78
ANNALS OF THE MISSOURI BOTANICAL GARDEN
TABLE IX
COLLECTION AND EXAMINATION
No. of experiment 1
2
3
4
5
6
7
3/22/13
Date of collection
2/12/13
2/22/13
3/15/13
3/15/13
3/19/13
3/22/13
Sampling place
Lab.
Outdoors
Street cai
5 and
10c. store
Street car Street cai
5 and
10c store
Temperature C*FO
Outdoors
60
29
32
36
50
- ■ ■
23
50
Sampling pi.
80
29
45
70
48 1 45
70
Weather conditions
Sunshine
Snowing
Windy
snowing
Windy
snowing
Sunshine
Sunshine
Sunshine
Approx. no. of
Sitting
44
1
44
44
1
persona
■
Standing 2
100
30
30
50
Approx. volume of sampling pi.
(cu. ft.)
3000
2500
72000
2500
2500
F
72000
Volume of air exam, (co.)
7800
22500
10800
10800
10800
10800 10800
Rate of filtration Ccc. per min.)
520
750
900
900
900
900
900
PI. I. (1 cc.)
2
2
4 30
9
250
i
PL II. (1 CO.)
2
8 3
36
6
250
1
3
No. of organisms
in following
quantities of
sand solution
plated
1
PL III. (2 cc.)
1
4
i
PL IV (5 cc.)
6
12
230
25
1
i
Too num-
erous to
count.
Spreader
PL V. (5 CO.)
1
1
20
200
1
1
1
1
1
26
1
Too num-
erous to
count.
23
Plates examined
I., II. and
III.
1
I., II. and
IV.
V.
II.
V.
1
1/12 of I. V
Total col. on plates exam.
8
16
20
36
26
21 on 1/12
of L
23
No. of bacilli, molds, etc.
4
8
d
16
16
5 on l/l2
of I.
15
No. of cocci
. .
4
8
11
— ■ - *- ■
20
10
16 on l/l2
of I.
8
No. of salivary cocci
1
2
1
6 on l/l2
of I.
Frequency of occurrence
1 in 1950
1 in 1800
1 in 10
Sanitary quality
Probably
unsafe
Safe
Probably
unsafe
Safe
Safe
Unsafe
Safe
No. of org. in total vol. of air
30
42
50
570
95
3750
58
1914]
NOLTE — SALIVARY ORGANISMS AND AIR POLLUTION
79
TABLE IX {Continued)
DATA ON THE COLLECTION AND EXAMINATION OF AIR SAMPLES
No. of experiment
8
9 10 11
12
13
14
Dato of collection.
3/29/ 3
, 3/29/13
4/1/13
4/1/13 '
4/8/13
4/9/13
4/10/13
Sampling place
Street car
Picture
show
Street car
picture
show
Outdoors
1
Outdoors
Outdoors
1
i
Temperature'(**F.)
Outdoors
41
59
50 77 45
63
65
1
SampUng pi.
55
1
1
70 ^ 63
82
]
45
63
55
Weather conditiona
Sunshine
Cloudy
1
Sunshine
Sunshine
Rain,
very
windy ;
Cloudy
Sunshine,
very
windy
Approx. no. of
Sitting
44
3000
44
3000
*
persons
Standing
30
100
15
300
1
1
1
Approx. volume of sampling pi.
(cu. ft.)
2630
90000
2500
— ,
90000
— .
Volume of air exam, (cc.)
10800 10800
10800 10800
10800
!
10800 10800
Rate of filtration (cc. per min.)
900 900
1
900
900
900
830 1080
J
PI. I. (1 cc.)
34 3
4
7
9
2
No. of organisms
PL 11. (1 cc.)
18 2
1
2
3
3
2
in following
quantities of
PL III. (2 cc.)
24
Spreading
mold
i
sand solution
plated
PL IV. (5 cc.)
14
Spreader
Spreader
1
1
1
1
1
3
PL V. (5 cc.)
8
12
' 35*
6
1
!
1
Plates examined
I.
TV.
V.
I. and II.
I., II. and
V.
IV. and V.
.I.,II.JV.,
andV-
Total col. on plates exam.
34
14
12
10
18
2
8
No, of bacilli, molds, etc.
15
12
6
9
9
2
6
No. of cocci
19
2
7
1
1 9
3
No. of salivary cocci
1
1
1
7
Frequency of occurrence
1
1 in 3600
1 in 1235
w. 1
Sanitary quality
1
Safe
Safe
Question-
able
Safe
t
Safe
Safe
No. of org. in total vol. of air
320
35
1
42
85
1
50
J
2
18
* Abundance of molds.
t Local contamination.
6
80 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1, 1914]
the salivary coccus form could not be found in the open air
devoid of the immediate presence of human beings.
It thus appears that the presence of the salivary coccus form
in air indicates the presence of man; furthermore, it indicates
the pollution of air by particles of mucus from the mouth.
Fliigge ^ and his school have shown that pathogenic organisms
may be transmitted into the air, and other workers ^ have shown
that the tubercle organism is capable of being carried by even
such feeble air currents as ordinarily exist in dwellings.
The tubercle organism, as well as the characteristic salivary
organism, is present in the saliva of tubercular patients. If,
therefore, this salivary organism can be isolated from the air
by means of the filter used in the above experiments, does it
not follow that the tubercle organism could be isolated in a
similar way? Since our manner of breathing is comparable
to the operation of the apparatus used, it follows that the tuber-
cle organism may be inhaled by man.
It thus appears that the presence in the air of the most char-
acteristic salivary organism is an index of the possible access
of pathogenic organisms to the atmosphere.
In conclusion, the writer wishes to express his thanks to Dr.
Geo. T. Moore, for valuable suggestions and numerous courte-
sies extended during the progress of the work; to Dr. J. R.
Schramm, for suggestions, and aid in the preparation of the
manuscript; and to Mr. Wilson F. Monfort, Chemist of the
City of St. Louis Water Department, for advice given and op-
portunities provided for the collection and examination of the
samples.
Explanation of Plate
PLATE 2
exhaust
suction tubes, and pressure bulb.
1
Gordon, M. H. loc. cil.
assermann, Handbuch der pathogenen Mikroorganisme
' - $
■\
Anx. Mo. Bot. Gard.. Vol. 1. 1914
Plate 2
f
•1
u
K
t
*
9
»
3
i
>,
\
rf - J
»r4^.
NOLTE— SAL1\ ARY ORGANISMS AND AIR POLLUTION
THE POLYPORACE^ OF OHIO
L. 0. OVERHOLTS
Rufus J. Lackland Fellow in the Henry Shaw School of Botany of
WashtTigton University
Introduction
I
I
h
The Polyporaccce, or "pore fungi," constitute a relatively
small family of the Basidiomycetes, characterized by having
the spores borne on the interior surfaces of tubes or pores which
make up the hymenium of the fungus. In its most compre-
hensive sense the family embraces the two subfamilies Boletece
and Polyporece, including also such aberrant genera as Merulius,
Porothelium, Solenia, etc. More often the Boletece are m.ade a
separate family, the Boletacece, usually distinguished from the
true Polyporacece by the more fleshy nature of the plant and by
the fact that the pores rather easily separate in a smooth layer
from the flesh of the pileus. The true Polyporacece, on the
other hand are more commonly leathery, corky, or woody in
texture, and only in rare cases are the tubes separable from the
context. More recently Dr. Murrill, who has monographed the
North American species of the family for the North American
Flora — now being issued by the New York Botanical Garden — ,
has still further limited the family so as to exclude not only the
genera referred to above, but also certain of the true polypores
which possess a more or less gelatinous or waxy hymenium. For
the reception of certain of these forms he has erected the family
Xylophagacece.
C. G. Lloyd has published monographic papers on certain of
the sections of the family, using for the most part as the generic
names, the sectional names given by Fries. Within the past-
year a third system of classification has been proposed by Miss.
Ames, of Cornell University, who divides the family into groups;
on the character of the context, and these groups are separated
into genera on the form of the fruit body, surface modifications,
spore characters, etc. Various workers in Europe have at-
Ann. Mo. Bot. Gabd., Vol. 1, 1914
(81)
[Vol. 1
82 ANNALS OF THE MISSOURI BOTANICAL GARDEN
tempted to revise the genera of the Polyporaceoe but none of
these classifications have been generally adopted by mycologists.
The family is here taken to include the following genera:
Polyporus (including P oly stictus) , Fomes, Trametes, D(sdalea,
Lenzites, Cyclomyces, Favolus, Gloeoporus, Merulius, and Irpex.
Distributed among these genera are practically one hundred
species found within the state. Of these, 78 have been col-
lected by the writer,4 others have been sent in by correspondents,
and examination has been made of collections of 5 other species
taken within the state and preserved either at the Lloyd Mu-
seum at Cincinnati, or in the herbarium of the New York Botan-
ical Garden. Of the remaining 12 species some are known only
from the records left by Morgan, Lea, Montague, Berkeley,
and Kellerman, others are admitted because there is every reason
to believe that they will be found within the state since they
are known to have been collected in nearby counties of adjoining
states.
The resupinate Polyporacece, usually included in the genus
Porta, have been omitted from this paper. Very little is known
in this country concerning these forms and very few authentic
specimens were available for study and comparison. Most of
the species that have been reported from this country have
been based on scarcely more than a guess, and it is impossible
for the amateur mycologist to determine his material from the
confused and often fragmentary account that has been written.
Until the genus has been thoroughly studied by a competent
mycologist, only added confusion would result from anything
more than a reference to it in this paper.
In the preparation of the keys, relationships, both of genera
and species, have been entirely ignored, the aim being to produce
a usable key rather than to exhibit relationships. The writer
beUeves that the color of the context is one of the most constant
of the gross characters of these plants, and the genera are divided
into sections on that basis. The presence or absence of a stipe,
the duration of the plant, the hymenial configuration, the
surface markings of the pileus, etc., are brought into the key
in an order which the writer believes corresponds to their rela-
tive importance as specific distinguishing characters. Spore
r.VoT.or.fov« Pf=!r»ppi'illv snore colors, are not used in the separation
1914]
OVERHOLTS — THE POLYPORACEjE OF OHIO 83
Sim
of the genera, and in the separation of the species only where
experience has shown that the spores are always easily obtained.
In mauy cases it is impossible to obtain spores, especially if they
be uncolored, from the hymenium of dried plants. However,
when plants are taken in the fresh condition it is usually a
)le matter to obtain them by leaving the fungus over
night in a moist atmosphere and allowing the spores to fall upon
a glass slide. Spores of the perennial woody forms may often
be obtained by this method when an examination of the same
material in the dried state does not reveal their presence. In
this paper spore measurements have been freely taken from
other pubUcations, both European and American. This was
done in order that the descriptions might be made more com-
parable. Due credit is given to the author in every case where
this was done.
An effort has been made to make the descriptions exactly
comparable one with another. For this purpose a definite
sequence of presentation has been arranged for the different
characters and this order preserved in all but a few instances
I
in which entire descriptions were taken from the original sources.
In the comments following each species the characteristic
specific distinctions are pointed out and references are made to
illustrations of one sort or another that give a good idea of the
plant as the writer understands it. Practically all of these
references are to papers published in this country. The writer
has had access to all of the important publications on the family,
both European and American. Most of the European writings
are not available to a large part of those students for whom this
paper is intended and it was believed that a careful selection of
citations to the illustrations published in this country would
be of more value than citations to the less known and often inac-
cessible European publications. Those who are in a position
to look up additional references will have access as well to
volumes 19 and 20 of Saccardo's 'Sylloge Fungorum,' where an
exhaustive index to illustrations will be found.
It is beheved that there can be no question of the need of a
paper worked out along the above indicated lines. No such
publication exists for any state in the Union and the only aids
that students have had in determining their collections have
[Vol. 1
84 ANNALS OF THE MISSOURI BOTANICAL GARDEN
been either the incomplete ''mushroom" books or such extensive
works as 'Sylloge Fungorum' and in more recent years the mono-
graph presented in the 'North American Flora.'
In the matter of citation and nomenclature an attempt has
been made to follow the rules and recommendations of the Inter-
national Botanical Congress at Brussels. Since there has been
little opportunity to compare specimens of our plants with those
of Europe or with type specimens, the procedure in the matter
of synonymy has been very conservative. The only names
cited as synonyms are those of which the writer has a personal
knowledge gained from the examination of authentic material,
usually species described from Ohio. Where there has been a
doubt as to the identity of a plant in this country with that of
one in the old world the procedure has been to use the name
under which it has been described or known in this country.
The first and therefore the most complete set of specimens is
in the herbarium of the writer; a set of all of the more common
forms is in the herbarium of Dr. Bruce Fink, of Miami Univer-
sity, at Oxford, Ohio ; a partial set is in the state herbarium, at
Columbus; and a large number of species, sent to Dr. Murrill for
determination and verification, are in the herbarium of the New
York Botanical Garden.
The writer is under deep obligations to the following persons
in various ways: First of all to Dr. Bruce Fink, under whose
direction the work was begun, whose aid, criticism, and advice
has made this publication possible; to Dr. W. A. Murrill, of the
New York Botanical Garden, for many kindnesses in verifying
and determining specimens sent to him, and for the privilege
of studying the specimens in the herbarium at that place; to
Mr. C. G. Lloyd, of Cincinnati, for the privilege of working in
the Lloyd Library and Museum and for determinations of speci-
mens; to Rev. G. Bresadola, of Trient, Tyrol, for determination
of specimens; to Dr. E. A. Burt, of the Missouri Botanical
Garden, for access to his herbarium and for suggestions as to the
final form of the paper; and to all who have aided in the work
by sending specimens and in various other ways.
It is hoped that the paper will be found useful not only to
Ohio students but in the neighboring states of the Great Lakes
1§U]
OVERHOLTS^ — THE POLYPORACE-E OF OHIO 85
region and in the Ohio valley as well. It is with this idea
mind that the paper has been prepared.
Key to the Genera.
Sporophore entirely resupinate; pilous none Poria^
Sporophore sessile or stipitate, sometimes effused-reflexed but not nor-
mally entirely resupinate
1
1. Hymenium composed of concentric lamellae; pileus stipitate. .Cydomyces p, 147
1. Hymenium not composed of concentric lamellae; pileus sessile or stipitate ... 2
2. Hymenium not distinctly poroid, the pores reduced to shallow pits sep-
arated by narrow ridges or reticulations Merulius p. 150
2. Hymenium distinctly poroid, irpiciform^ dsedaloid or lamellate, but not
pitted 3
3. Hymenium more or less waxy or gelatinous, the layers of tubes separating
smoothly from the context in fresh specimens or when moistened;
pileus sessile, thin and flexible Gloeoporus p. 149
3. Hymenium not at all waxy or gelatinous and not separating smoothly
from the context; pileus sessile or stipitate
4
4. Hymenium either dsedaloid, labyrinthiform or lamellate, at least in part 5
4, Hymenium poroid or sometimes broken up into teeth
6. Context white 6
5. Context brown 8
6. Pileus minutely velvety to glabrous; context more than 1 mm. thick
Dcedalea p. 143
6. Pileus hirsute to villous; context 1 mm. or less thick 7
7. Hymenium lamellate, at least in part Lenzites p. 145
7. Hymenium dsedaloid but never lamellate Dcedalea p. 143
8. Plants woody and perennial, more than 1 cm. thick; hsnmenium not at
all lamellate Trametes p. 138
8. Plants coriaceous or corky, less than 1 cm. thick; hymenium often
lamellate Lenzites p. 145
10
. 13
9. Hymenium broken up into teeth
9. Hymenium poroid, not broken up into teeth
10. Tubes or teeth 5 mm, or more long Irpex p, 151
10. Tubes or teeth less than 5 mm. long 11
11. Hymenium labyrinthiform at first and remaining so at the margin
11. Hymenium never labyrinthiform
Dcedalea p. 143
12
12. Pileus less than 1 cm. broad; fungus mostly resupinate Trpex p. 151
12. Pileus more than 1 cm. broad; fungus not mostly resupinate
Polypoms p.
A
13. Pores large and hexagonal; stipe present
14
13. Pores small and circular or angular; stipe present or absent 15
14. Stipe lateral, often rudimentary; pores usually radiating and longer in
the radial direction Favolus p. 148
14. Stipe usually central or subcentral; pores not radiating Polypoms p.
16
15. Tubes in a single layer; plants annual
16. Tubes in two to several layers; plants perennial
r
»See Introduction p. 82.
17
[Vol, 1
86 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
16. Tubes not in a distinct stratum but appearing to be simken to differ-
ent depths into the context Trametes p. 138
16. Tubes forming a well marked stratum entirely distinct from the
^^^^^t • Polyporus p. 86
17. Hymenium bright yellowish brown; plants growing only on the wood
of coniferous trees Trametes p. 138
17. Hymenium whitish, flesh-colored, dull brown, etc., but not bright yel-
lowish brown; plants growing on the wood of either coniferous or decid-
uous trees p^^^ v. 126
Descriptions and Keys to the Species
years,
POLYPORUS Mich, ex Fries,
Syst. Myc. i: 341. 1821; Mich. Nov. Plant. Gen. 129. 1729.
Plants annual or in rare cases persisting for two or three
terrestrial or epixylous, sessile or stipitate; pileus fleshy,
coriaceous or corky in texture, small or of immense size, often
brightly colored; context white, yellow, red, or brown; tubes in
a single layer, all sunken into the context to an equal depth so
that their bases form a definite continuous straight line; mouths
mostly circular or angular, in rare cases showing a favoloid or
daedaloid tendency and sometimes breaking up into teeth;
stipe (when present) variable in position and texture; spores
white (bluish in one species), or some shade of brown.
KEY TO THE SPECIES
Context white or whitish Section I
Context reddish or yellowish Section II
Context brown or brownish Section III.
Section I,
Sporophore stipitate or substipitate
1
Sporophore sessile or sometimes effused-reflexed but never stipitate 21
1. Pileus and stipe covered with a reddish varnish 2
1. Pileus and stipe not red-varnished 3
2. Varnish disappearing with age, the pileus then whitish or yellowish
61. P. Curiisli
2. Varnish persisting, the pileus not changing color 60. P. lucidus
3. Plant small, not more than 1 cm. high 99. P. mcvla
3. Plant always much larger 4
4. Stipe compound, branching near the base; pileoli usually several or many . 5
4. Stipe simple or not branching more than once; pileus generally single. ... 9
5. Pileoli small (usually less than 5 cm. broad) and numerous 6
6. Pileoli large (5 cm. or more broad) and fpw in number 7
V
II
i
1914]
OVERHOLTS — THE POLYPORACEJS OF OHIO 87
6. Pileoli regular in outline and centrally attached; the branches of the stipe
regular and cylindrical in form 35. P. umbellatiLS
6. Pileoli always laterally attached; the stipe branches irregular. 39, PJrondosvs
7. Spores roughly echinulate P- P- Berkeleyi
7. Spores smooth 8
8. Pileus pallid or light brown; hymenium usually turning black where
bruised and on drying 40. P. giganteus
8. Pileus yellowish green; hymenium not turning black 57. P. flavovirens
9. Context soft and spongy above, firm next to the hymenium; plants often much
distorted; usually growing about stumps 38, P, dislorlus
9. Context uniform; plants not distorted 10
10. Plants growing on the ground H
10. Plants growing on wood 12
11. Stipe black and rooting at the base; pileus some shade of brown. .36, P. radicatus
11. Stipe not black and rooting at the base; pileus yellowish green. .37. P. fluvomrens
12. Sporophore more or less globose; tubes concealed by a volva. .^7. P. volvulus
12. Sporophore not globose; volva absent 13
13. Sporophore arising from a cup-shaped, sterile body that sometimes disap-
pears; pileus white; found only on dead branches of Ulmics 6. P, conchifer
13. Sporophore not arising from a cup-shaped sterile body 14
14. Margin of the pileus projecting 6 mm. or more beyond the hymenium;
hymenium separating smoothly from the context in fresh specimens;
I growing only on Betvia 36. P. betvlinxis
14. Plants not as above -. 15
15. Hymenium bright sulphur-yellow. 4^- P. snlphureus
15. Hymenium not bright sulphur-yellow ' 16
16. Mouths of the tubes minute, averaging 4-7 to a nim 17
16. Mouths of the tubes larger, averaging 1-3 to a mm 18
17. Mouths of the tubes averaging 4 to a mm.; pileus rarely more than 5 cm. in
diameter 55. P. elegans
17. Mouths of the tubes averaging about 6 to a mm.; pileus 4-20 cm, in diameter
34. P. picipes
18. Pileus large, more than 5 mm. thick; plant growing on living trees; stipe
black at the base 33, P. squanwsus
18. Pileus small or medium sized, not more than 5 mm. thick; stipe not
black at the base 19
19. Tubes long-dccurrent on the stipe; context soft and friable when dry
32. P. pennsylvanicus
19. Tubes slightly or not at all decurrent; context not soft and friable when dry . , 20
20. Pileus yellowish brown; mouths of the tubes almost 1 mm. in diameter;
walls thin SI. P. arcularius
20. Pileus darker than above, sometimes sooty-black; mouths of the tubes
averaging 2 to a mm.; walls at first thick 30. P. hrumalis
21. Pileus red-varnished, at least when young
22
21. Pileus never red-varnished 23
22. Varnish disappearing with age, the pileus then whitish or yellowish
61. P. Curtisli
22. Varnish persistent, the pileus not changing color 60. P. lucidus
23. Sporophore more or less globose; tubes concealed by a volva 27. P. volvatus
23. Sporophore not globose; volva absent 24
[Vol, 1
88 ANNALS OF THE MISSOURI BOTANICAL GARDEN
24. Sporophore arising from the under side of a cup-shaped, sterile body;
found only on dead branches of Ulmus 6, P, conchifer
24. Sporophore not arising from a cup-shaped, sterile body 25
25. Margin of the pileus projecting 5 mm. or more beyond the hymenium;
hymenium separating smoothly from the context in fresh specimens; found
only on Betula ^g. p, bciulinus
25. Plants not as above 26
26. Hymenium bright sulphur-yellow ^^. P. sulphtireus
26. Hymenium not bright sulphur-yellow 27
27. Pileus distinctly browTi in color; context usually light browTi; hymenium
changing color when bruised 46, P.
resmosus
27, Pileus not brown in color; hymenium never changing color when bruised . . 28
28. Hymenium more or less smoke-colored or black 29
28. Hymenium not at all smoke-colored or black 32
29. Pileus more than 4 mm. thick 30
29. Pileus not more than 4 mm. thick 31
^ 30. Context fragrant, with the odor of anise S3, P. fragrans
30. Context not fragrant, odor sometimes disagreeable 24, P.furnosus
31. Mouths of the tubes angular, minute, averaging 5-7 to a mm.; dissepiments
*^^n S2, P. adustus
31. Mouths of the tubes circular or subcircular, medium sized, averaging 3-5 to a
mm.
umosus
; dissepiments thick ^ 24, p. /
32. Context fibrous or coriaceous in fresh plants; pileus never more than 1.5
cm. thick, and usually much thinner 33
32. Context either soft, spongy and ful! of water or firm and corky, often
fragile when dry; pileus often more than 1.5 cm. thick 43
33. Hymenium broken up into teeth ; 34
33. Hymenium entire or lacerate but not broken up into teeth 36
34. Context more than 1 mm. thick 9, P, hijormis
34. Context not more than 1 mm. thick 35
35. Plants growing only on the wood of coniferous trees 2.P, abietinus
35. Plants growing only on the wood of deciduous trees 3, P, pargamenus
36. Context 1 mm. or less thick 37
36. Context more than 1 mm. thick 40
37. Mouths of the tubes minute, averaging 4^6 to a mm. ; hymenium never violet
or purple 38
37. Mouths of the tubes larger, averaging 2-3 to a mm.; hymenium often violet
or purple 39
38. Surface of the pileus villous or velvety; pileus multizonate, generally
more than 2 cm. broad 1. P. versicolor
38. Surface of the pileus densely hirsute; pileus azonate or with one or two
zones, generally less then 2 cm. broad 4. P, hirsutuLus
39. Plants growing only on the wood of coniferous trees 2. P, abietinus
39. Plants growing only on the wood of deciduous trees 5. P. pargamenus
40. Mouths of the tubes large, averaging 1-2 to a mm 9, P. Uformis
40. Mouths of the tubes medium sized, averaging 3-4 to a mm 41
41. Tubes more than 2 mm. long 7, P. pubescens
41. Tubes not more than 2 mm. long 42
42. Surface of the pileus velvety to hirsute 5. P. hirsvius
42. Surface of the pileus minutely pubescent or glabrous 8. P. Lloydii
}
1914]
OVERHOLTS — THE POLYPORACEiE OF OHIO 89
43. Plants mostly resupinate 44
43. Plants not mostly resupinate 45
44. Pileus azonate, margin often inrolled 10. P. semipileatiis
44. Pileus zonate, margin always straight SL P zonalis
45. Pileus corky in texture when fresh, usually rather thick and firm 46
45. Pileus soft and spongy in texture when fresh 49
46. Pileus distinctly encrusted; hymenium and context pinkish or rosy when
fresh; plants usually growing on Fraxinus P. fraxineus^
46. Pileus not encrusted; hymenium and context whitish when fresh; plants
not usually on Fraxinus 47
47. Pileus more than 2 cm. thick; tubes more than 4 mm. long 25, P. robiniophila
47. Pileus not more than 2 cm. thick; tubes not more than 4 mm. long 48
48. Plants with a sweet anise odor M. P.fragrans
48. Plants with no odor, or odor disagreeable S4^ P, fumosus
49. Mouths of the tubes minute, averaging 6-7 to a mm.; plants with a sweet
acid odor I4, P. galcwtinus
49. Mouths of the tubes larger, averaging 1-4 to a mm 50
50. Pileus generally less than 4 cm. broad 51
50. Pileus generally more than 4 cm. broad 53
61. Pileus pubescent; mouths of the tubes dentate, lacerate, or irregular 52
61. Pileus glabrous; mouths of the tubes entire; plants with a sweet acid odor
12. P. chioneuB
62. Pileus and spores (in mass) often bluish or slate-colored; tubes equalling
in length the thickness of the context 11. P. caesius
52. Pileus and spores pure white; tubes shorter in length than the thickness
of the context 13, P, lacteus
53. Plants growing only on the wood of coniferous trees 54
63. Plants growing only on the wood of deciduous trees 55
64. Tubes usually more than 5 mm. long, the mouths averaging 2-3 to a mm.
19. P. horealis
54. Tubes usually less than 5 mm. long, the mouths averaging 4-5 to mm,
18, P. guttidatus
65. Margin of the pileus thick and rounded 17. P. ohtusus
65. Margin of the pileus thin and acute 56
56. Mouths of the tubes large, averaging 1-2 to a mm 16. P. deledans
56. Mouths of the tubes small, averaging 3-5 to a mm. . . . ; 57
57. Fresh plant with a disagreeable odor; context very hard when dry . 20. P. Spraguei
67. Fresh plant with no disagreeable odor 15. P, spumeus
Section II
Pileus and hymeniimi deep cinnabar-red .'
Pileus and h^nnenium not deep cinnabar-red (rosy or orange-colored in some
1
species) 2
1. Pileus less than 5 mm. thick, often zonate " 44- P- sanguineus
1. Pileus more than 5 mm. thick, never zonate 4^. P. cinnaharinus
2. Hymenium bright sulphur-yellow in fresh plants 4^. P. sulphureus
2, Hymenium not bright sulphur-yellow 3
8. Plant growing only on the wood of Qu£rcm and Castanea; pileus yellowish or
ge-colored 43^ P. PHotce
oran
I For description see p. 130 under the genua Fomes.
90 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
3. Plant growing usually on Fraxinus; pileus usually stained more or less with
^^ P. fraxineus*
3. Plant growing usually on coniferous wood; rose-colored without and within . , .
P. cameus*
Section III
Pileus stipitate or substipitate 1
Pileus sessile or efTused-reflexed, not stipitate Q^
1. Pileus and stipe covered with a rcxidish varnish at least when young 2
1. Pileus and stipe not red- varnished 3
2. Pileus and stipe at first red-varnished, the varnish disjippearing and the
pileus becoming whitish or yellowish when mature 61. P, Curtisii
2. Pileus and stipe strongly red- varnished, the varnish not disappearing with
age 60, P, lucidus
3. Context not more than 1 mm, thick; plants growing on the ground ........ 4
3. Context more than 1 mm, thick; plants growing on wood or attached to buried
wood
d
4. Surface of the pileus marked with silky striations 59. P. cinnamomeus
4. Surface of the pileus not silky 5
5. Mouths of the tubes small, averaging 2-4 to a mm.; tubes usually less than 3
^^- long 5s. P, perennis
5. Mouths of the tubes large, averaging 0.5-1 mm. or more in diameter; tubes
usually more than 3 mm. long 57, P, focicola
6. Surface of the pileus distinctly encrusted P, lobatus^
6. Surface of the pileus not at all encrusted 7
7. Context decidedly duplex, spongy above, firm next to the tubes . . .65. P. circinatus
7. Context not duplex g
8. HjTnenium some shade of yellow (yellowish brown, yellowish green, etc.),
quickly changing color when bruised; growing on or about trees and
stumps of Pi7ius; spores white 54. P. Schweinitzii
8. Hymenium cinereous to brownish, not changing color when bruised;
growing on the ground or attached to buried wood; spores brown
66. P. obesus
9. Pilei forming a densely imbricate, globose or cylindrical mass P. graveolem*
9. Pilei not forming a densely imbricate, globose or cylindrical mass 10
10. Pileus red-varnished 60. P. lucidus
10. Pileus not red-varnished H
11. Pileus distinctly encrusted p. lobatus*
11. Pileus not distinctly encrusted 12
12. Plants growing on or about stumps or trunks of Pinus. . .64. P. Schweinitzii
12. Plants growing on wood of deciduous shrubs or trees, often on living
trunks I3
13. Context usually less than 7 mm. thick; plants small or medium sized 14
13. Context more than 7 mm. thick; plants large 17
14. Spores white I5
14. Spores brown 1q
>See p. 130 for a description of this plant.
»For description of this plant see p. 131 under the genus Fomes.
•This plant is described on p. 137 under the genus Fomes.
*FoT description see p. 131 under the genus Fomes.
*For description see p. 137 under the genus Fomes.
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 91
15. Pileus spongy and watery when freah; context friable when dry; mouths of
the tubes averaging 2-4 to a mm 47. P. nidulans
15. Pileus firm and rigid; context corky when dry; mouths of the tubes minute,
averaging 5-8 to a mm 4^. P. gilvus
16. Plants growing on the wood Of Alnus and Betula; spores light brown. . .
49. P. radialus
16. Plants growing on the wood of Acer, Fagus, and other deciduous trees . . .
60. P. cuticularis
17. Context very light brown 46. P. resinosus
17. Context yellowish brown or darker 18
18. Surface of the pileus hirsute; plants growing on various diseased decid-
t,rees 61. P. hispidus
uous
18. Surface of the pileus fibrillose or glabrous; plants growing only on the
wood of Quercus 19
19.' Sporophore medium sized, less than 10 cm. broad and 3 cm. thick
63. P. dryophilus
19. Sporophore large, more than 10 cm. broad and 3 cm. thick 62. P. dryadeus
I. P. versicolor L. ex Fries, Syst. Myc. i: 368. 1821.
Boletus versicolor L. Sp. Plant. 1176. 1753.
Pileus sessile or effused-reflexed, imbricate or single, dimid-
iate or encircling twigs and then often orbicular by confluence,
2-5 X 2-7 X 0.1-0.3 cm., coriaceous, prevailing color grayish,
but marked by many narrow, multicolored zones, ranging
from white to yellow, brown, reddish, greenish, blackish, etc.,
villous or velvety, the margin thin and acute, usually sterile
below; context white or whitish, fibrous, less than 1 mm. thick;
tubes 1-2 mm. long, the mouths white or yellowish, sometimes
somewhat glistening, circular to angular, averaging 3-5 to a
mm., the walls thin, entire or slightly lacerate; spores white,
smooth, oblong, sometimes curved, 1.2-2 x 5-6.3 m.
On all kinds of dead wood. Common throughout the year.
Easily distinguished by the multizonate, multicolored pileus.
P. hirsutulus Schw. is often considered to be a form of this
species. P. zonatus Fries, as reported by Morgan, is one of the
many forms of it. The following references contain good illus-
trations of our plant: Hard, Mushrooms/. 343., White, Hymen.
Conn, pi 36., and Moffatt, Higher fungi of the Chicago region
pin.f.L
2, P. abietinus Dicks, ex Fries, Syst. Myc. i : 370. 1821.
Boletus abietinus Bicks. Yasc. PI Crypt. Brit 3:21. 1793.
Pileus sessile or effused-reflexed, dimidiate and broadly
attached, or flabelliform and attached by the attenuate base
[Vol, 1
92 ANNALS OF THE MISSOURI BOTANICAL GARDEN
of the pileus, 0.5-5 x 0.5-5 x 0.1-0.2 cm., coriaceous, white to
cinereous or almost black behind, villous, zonate, margin thin
and acute ; context white or pallid, fibrous, not more than 1 mm.
thick; tubes less than 3 mm. long, the mouths white to bay and
often violaceous toward the mardn. averairine- 1-?, t,n n
mm
the dissepiments thin and soon lacerate and breaking up into
teeth
Growing only on the wood of coniferous trees. In autumn.
Rare.
Closely related to P. pargamenus Fries, from which it is
most easily separated by the habitat. The following spore
dimensions are found in the literature: Karsten—" oblong 4-6
X 1-3 /i"; Murrill— "globose, smooth, hyaline, 4.5-5.5 /x in diam-
eter"; Bresadola— "hyaline, cylindrical, subcurved, 6-7 x 2.5 a^."
3. P. pargamenus Fries, Epicr. Syst. Myc. 480. 1838.
Pileus sessile or effused-reflexed, imbricate, dimJdiate or
flabelhform, sometimes attached by an attenuate base, 1-7
X 1-7 x 0.1-0.4 cm., coriaceous, whitish to cinereous or yellowish
brown, villous, zonate, the zones sometimes differently colored,
margin very thin, acute, broadly sterile below, often violaceous
in color; context white or whitish, fibrous, very thin, less than
1 mm. thick; tubes not more than 2.5 mm. long, the mouths
whitish to bay and often violaceous toward the margin, angular,
averaging 2-3 to a mm., the dissepiments thin and soon break-
ing up into teeth; spores white, smooth, oblong, slightly curved
2-2.5 X 5-6.3 n.
Growing on the wood of deciduous trees, especially of Quercus
and Prunus. September to December. Common.
Close to P. abietinus Dicks, ex Fries, but usually found on
'dead wood of deciduous trees. Well represented by Hard
(Mushrooms /. 345) as P. pergamenus.
4. P. hirsutulus Schw. Trans. Am. Phil. Soc. II. 4: 156
1832.
Pileus sessile or effused-reflexed, often imbricate, dimidiate,
0.5-2 x 0.5-2.7 X 0.1-0.2 cm., coriaceous, gray or cinereous to
yellowish brown, hirsute or strigose, azonate or with 2-3
colored zones, margin thin and acute, usually sterile below;
context white or whitish, membranous, less than 1 mm. thick;
tubes less than 2 mm. long, mouths whitish to vellowish, rarelv
1914]
OVERHOLTS — THE POLYPOBACE^ OF OHIO 93
glistening, circular or angular, averaging 3-5 to a mm., the
dissepiments thin and entire.
On dead branches of deciduous trees, more often on fruit
trees. Found from August to December. Not common.
Separated from P. versicolor L. ex Fries, by the more hirsute
or strigose pubescence on the pileus, and by the smaller size.
Specimens collected at Cincinnati by D. L. James and referred
to P. velutinus Fries are now referred to this species.
5 P. hirsutus Wulfen, ex Fries, Syst. Myc. i : 367. 1821.
Boletus hirsutus Wulfen, in Jacq. Coll. 2: 149. 1788.
Pileus sessile, or effused-reflexed, dimidiate, 1.5-5 x 1.5-7
X 0.2-1 cm., flexible when moist, firm and sometimes rigid
when dry, grayish to yellowish or smoky brown, hirsute or
tomentose, sometimes zonate, sometimes concentrically sulcate,
the margin thin or rather thick, acute, sometimes dark colored;
context white or pallid, tough to soft-corky, 1-6 mm. thick;
tubes 1-4 mm. long, the mouths white, grayish or fuliginous,
circular to somewhat angular, averaging 3-4 to a mm., the
walls rather thick and always entire; spores white, smooth,
cyhndrical, often curved, 2.5 x 5-8 /x.
On dead wood of deciduous trees. Found throughout the
year.
From closely related species with a conspicuous hairy covering
this plant is perhaps most easily separated by the persistently
thick walled tubes that never become torn or lacerate. Any
plant with the characteristics of this group and possessing the
dark-colored marginal band to which reference is made in the
description may always with safety be referred to this species.
From P. versicolor L. ex Fries, the plant is separated by the
absence of the numerous multicolored zones. Hard's figure
(Mushrooms /. 342) is not a good illustration of our plant.
Murrill describes the plant under the name of Coriolus nigro-
marginatus (Schw.) Murr.
6. P. conchifer Schw. ex Fries, Epicr. Syst. Myc. 463. 1838.
Boletus conchifer Schw. Syn. Fung. Car. 98. 1822.
Pileus sessile or attached by a lateral tubercle and then
appearing substipitate, reniform to dimidiate in outline, 1-3
X 1-4 X 0.1-0.3 cm., coriaceous, white to yellowish, glabrous,
[Vol. 1
94 ANNALS OF THE MISSOURI BOTANICAL GARDEN
zonate or azonate, the margin very thin and acute; on the
upper surface and at the base of the pileus a small cup-shaped
or disk-like sterile structure is usually borne, white or brown
and often zoned on the inside; context white, fibrous, less than
1 mm. thick; tubes not more than 2 mm. long, at first white,
often yellowish on drying, the mouths angular and thin- walled
averaging about 3 to a mm., the dissepiments often lacerate;
stipe (?) rudimentary, tubercular; spores not obtained.
Growing only on fallen branches of Ulmus. Common.
This plant has somewhat the appearance of P. puhescens
Schum. ex Fries, from which, however, it is easily separated
by the much thinner pileus, the attenuate base, the presence
of the sterile cup, and the habitat. The cup is sometimes
absent. The development of the cup has not been closely fol-
lowed. Lloyd believes that the fertile pileus is first developed
and from it the sterile cup arises, and that during the winter
the fertile portion falls away, the cup persisting on the sub-
stratum but not giving rise to new pilei the next season. Miss
Ames comes to the conclusion that the sterile cups represent
pilei whose marginal hyphse have been killed by unfavorable
conditions and which as a result may develop a fruiting surface
from the base of the dead cup-like pileus. This would explain
the occasional absence of the sterile cup, its presence depending
upon the death of the marginal hyphse in the early stages of
the production of a first pileus. P. virgineus Schw. described
from North Carolina is said to be this plant. The plant is
exceptionally well illustrated by Lloyd (Myc. Notes, Polyporoid
Issue 3 /. 365-66), and by Moffat (Higher fungi of the Chicago
region yl. 16. f. 2),
7. P. pubescens Schum. ex Fries, Syst. Myc. i : 367. 1821.
Boletus puhescens Schum. Enum. PI. Saell. 2: 384. 1803.
Polyporus Sullivantii Mont. Ann. Sci. Nat. II. 18: 243. 1842.
Pileus sessile, dimidiate, 1.5-5 x 2.5-5 x 0.4-1 cm., fleshy-
tough when fresh, firm when dry, white or yellowish in fresh
specimens, sometimes umber or brown when dry, villous-
tomentose, zonate or azonate, margin thin, acute; context
white or pallid, fibrous-tough when fresh, more firm when
dry, 1-5 mm. thick; tubes 1-4 mm. long, the mouths white,
yellowish, or umber, angular, averaging 3-4 to a mm., the
^
<
1914]
OVERHOLTS — THE POLYFORACEiE OF OHIO 95
dissepiments thin, entire to dentate; spores white, smooth,
^
cylindrical, curved, 2.7-3.6 x 5.4 n.
On dead wood of deciduous trees. August to November.
Common.
Plants collected in the Miami valley by Morgan and referred
by him to P. velutinus Fries belong here. Plants distributed
by Kellerman in his 'Fascicles of Ohio Fungi' as P. molliusculus
Berk, are referred to this species. P. fibula Fries as reported
by Morgan is probably the same as P. pubescens var. Grayii,
here included under P. pubescens. Hard (Mushrooms /. 339)
gives a good illustration of the plant.
8. P. Lloydii (Murr.) Overholts, n. comb.
Coriolus Lloydii Murrill, N. Am. Flora 9: 23. 1907.
Pileus rather thin, laterally connate, rigid, tough, cuneate
to flabelliform, applanate, tubercular-sessile, 2-3 x 3-4 x 0.2-0.4
cm.; surface white or isabelline, scabrous, somewhat rugose,
marked with a few narrow, indistinct, pale latericeous zones;
margin thin, fertile, irregular, lobed; context punky-fibrous,
white, 1.5-2 mm. thick; tubes 1-1.5 mm. long, white within,
mouths angular, subglistening, 4 to a mm., edges thin, firm,
dentate, white or isabelline; spores globose, smooth, hyaUne,
2 n\ hyphse 5 n.
On dead wood. Rare.
The above description is taken from the 'North American
Flora.' The type specimens were collected near Cincinnati,
Ohio, by C. G. Lloyd, and to the writer's knowledge the plant
has not been found since. The species appears to be distinct.
9. P. biformis Klotzsch, Linnaea 8: 486. 1833.
P. molliusculus Berk. Hooker's Lond. Jour. Bot. 6: 320.
1847.
Plants sessile, effused-reflexed or resupinate, often imbricate;
pileus dimidiate or laterally confluent and elongate, 0-5.5 x
1.5-6 X 0.2-1.5 cm., soft and phable when fresh, slightly
flexible to rigid when dry, white, palHd, bay, or ochraceous,
appressed-fibrillose, usually rough, azonate or subzonate, the
margin thin and acute; context white or whitish, fibrous-tough
when fresh, soft-corky when dry, 1-5 mm. thick; tubes white,
becoming bay on drying, 2-5 mm. long, the mouths circular
to angular or sinuous, averaging 1-2 to a mm., the dissepiments
7
4
%
[Vol. 1
96 ANNALS OF THE MISSOURI BOTANICAL GARDEN
rather thin and usually becoming lacerate and broken up into
teeth at an early stage of growth, sometimes remaining poroid,
especially toward the margin of the pileus; spores white, smooth,
oblong, curved, 2-2.6 x 7-8 /x.
Growing on old logs. September to December. Common.
The follow^ing group of characters will usually identify
the species: the semi-resupinate habit of growth, whitish or
tan-colored pileus, and the rather long tubes with large mouths,
soon breaking up into teeth. P. molliusculus was named by
Berkeley from specimens sent to him from Ohio by Lea. Mor-
gan's determination of P. molliuscuhis was an error, his plants
belonging to P. puhescens Schum. ex. Fries. Kellerman re-
peated the error in distributing P. molliusculus in his 'Ohio
Fungi Fascicles.' For illustration see Hard, Mushrooms/. 341.
10. P. semipileatus Peck, Ann. Rept. N. Y. State Mus.
34: 43. 1881.
Plants resupinate or effused-reflexed, rarely strictly sessile;
pileus dimidiate or elongate, 0-1.5 x 0.7-3.5 x 0.1-0.5 cm.,
soft and spongy when fresh, rigid when dry, white, yellowish,
or reddish brown, slightly tomentose to glabrous, azonate,
margin thin, acute; context whitish, soft when fresh, firm when
dry, 1-4 mm. thick; tubes less than 2 mm. long, the mouths
white, greenish or somewhat violaceous, angular, minute,
averaging 4-6 to a mm., the walls entire; spores white, smooth,
oblong, curved, 1 x 3-4 n.
On old limbs on the ground. September to December. Rare.
Easily recognized by the minute pores, the semi-resupinate
habit of growth, and the often violet tinted hymenium. There
is no previous record of the plant occurring in Ohio. Collec-
tions were made at Oxford, in 1911, for the first time.
11. P. caesius Schrad. ex Fries, Syst. Myc. i: 360. 1821.
Boletus caesius Schrad. Spic. Fl. Ger. 167. 1794.
Pileus sessile or effused-reflexed, dimidiate, 1-3.5 x 2-6 x
0.3-2 cm., soft and spongy when fresh, rigid when dry, whitish
to cinereous, often with a bluish tinge, distinctly villous or
tomentose especially behind; azonate, margin thin and acute;
context white, soft, spongy and full of water when fresh, friable
when dry, 0.3-1 cm. thick; tubes 2-7 mm. long, mouths w^hite,
pallid, or bluish gray, angular, averaging 3-5 to a mm., the
;
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 97
walls thin and usually lacerate; spores minute, white, smooth,
cylindrical, sometimes curved, 1.2-1.5 x 4.7-5.2 ju.
On dead wood of deciduous and coniferous trees. October
to December. Rare.
The bluish color of the pileus and hymenium is so often
wanting that other characters must frequently be used in the
identification of the plant. The slender tubes, usually longer
than or as long as the thickness of the context, is apparently
a rather constant character of the plant. The villous or tomen-
tose pileus separates it from P. chioneus Fries and P. lacteus
Fries and these are the only species with which it is likely to
be confused.
12. P. chioneus Fries, Syst. Myc. i: 359. 1821.
Pileus sessile, dimidiate, 1-3 x 2-5 x 0.5-3 cm., soft and
spongy when fresh, rigid when dry, whitish to grayish or yel-
lowish, azonate, glabrous or with a slight strigose tomentum
towards the base, sometimes covered wdth a thin grayish or
yellowish pellicle that becomes more evident on drying; margin
acute, sometimes inflexed on drying; context white, soft and
spongy when fresh, fragile when dry, 0.3-2 cm. thick, azonate,
with a sweet acid odor; tubes 1-8 mm. long, mouths white or
yellowish, usually glistening, angular, averaging 3-4 to a mm.,
the walls thin but entire; spores white, smooth, oblong, slightly
curved, 1-1.7 x 4-5 /x.
On dead wood. September to November.
From P. galaciinus Berk, this plant is most easily separated
by the oblong, curved spores. The usually glabrous pileus
and the absence of bluish tints separates it from P. caesius
Schrad. ex Fries. Whether it is distinct from P. lacteus Fries
may well be doubted. The plant is much in dispute in Europe,
Our plants have been described as P. alhellus Peck.
13. P. lacteus Fries, Syst. Myc. i: 359. 1821.
Pileus pure white, fleshy-fibrous, fragile, triangular, pubes-
cent, azonate externally and internally, margin inflexed, acute;
pores thin acute, dentate, becoming torn and labyrinthiform.
Commonly small and thin but sometimes large and transversely
elongate, often gibbous behind, becoming glabrate and uneven.
(Adapted from Fries, Hymen. Eur. 546.)
On dead wood of deciduous trees. Rare.
[Vol. 1
98 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Until quite recently this and the preceding species have been
held to be quite distinct. Of late years the European mycol-
ogists are coming to believe that they cannot be regarded as
distinct species. Murrill would separate them on the ground
that P. chioneus always has a distinct cuticle which is entirely
lacking in P. ladeus. The writer has endeavored to keep the
plants distinct on the basis of the differences noted by Fries.
If this proves unfeasible then the two must be united as one
species under the name of P. chioneus, at least with reference
to their occurrence in this country.
14. P. galactinus Berk., Hooker's Lond. Jour. Bot. 6:
321. 1847.
Pileus sessile, imbricate or single, dimidiate, 3-7 x 3-7 x 0.5-2
cm., soft and pliant when fresh, more or less watery, rigid and
contorted on drying, white, grayish, or somewhat yellowish,
tomentose to strigose-tomentose, especially at the base, be-
coming glabrous with age, azonate, margin thin and acute;
context white or pallid, watery and spongy when fresh, with
a distinct sweet acid odor, firm when dry, sometimes more
or less duplex, 3-8 mm. thick; tubes 2-7 mm. long, mouths
white to bay, often glistening, circular to angular or sinuous,
minute, averaging about 6 to a mm.; spores white, smooth,
ellipsoid, 2-2.5 x 3.5-4 /i, uninucleate and with a very trans-
parent wall.
Growing on dead wood of deciduous trees. August to
November. Common.
The sweet acid odor mentioned in the description is a dis-
tinguishing character of all collections of this species. No
mention is made of the odor in any published work to the
writer's knowledge, except in Peck's description of P. imviitus
in which the odor is described as subacid. P. immitus is in
all probability this plant. The odor is so constant that whenever
it is noticed in connection with any minute-pored form of this
section one can be sure that the plant belongs to this species.
All of the collections that I have referred to this species
are watery when fresh, have a sweet acid odor, and when dried
shrink much in size and often become much contorted. The
V
context becomes thin and hard and takes on a resinous, dark
brown or black color. This appearance may be uniform through
\
>
\
\
\
I
f
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 99
the context or the dark resinous color may be limited to a
narrow line next to the hymenium or confined to two or three
narrow zones in the context. It is difficult to distinguish these
species with a white watery context and the writer's presenta-
tion of them may be open to criticism.
15. P. spumeus Sow. ex Fries, Syst. Myc. i:358. 1821.
Boletus spumeus Sow. Col. Fig. Eng. Fungi pi. 211. 1797.
Pileus sessile, dimidiate, watery and fleshy-tough when fresh,
firm when dry, 7-12 x 10-20 x 2-3 cm., much smaller on dry-
ing, appearing appressed-tom.entose, white or grayish, some-
what yellowish or brownish on drying, azonate, margin rather
thick but acute; context white, soft, spongy, and full of water,
rather fragile on drying, more or less zonate, 1-3 cm. thick;
tubes 0.5-1.5 cm. long, mouths white or yellowish on drying,
angular, averaging 3 to a mm.; spores white, smooth, globose,
or subglobose, 4.5-5.2 ju in diameter, distinctly uninucleate.
Growing on injured or diseased deciduous trees, especially
Ulmus and Acer. October and November. Rare.
The plant is closely related to P. delectans Peck, with the
same habitat and general appearance, but separated from that
species by the smaller mouths of the tubes and by the distinctly
uninucleate and more globose spores. The plants so referred
do not agree with the figure given by Sowerby, nor with Fries'
description. My plants were determined by Bresadola.
i6. P. delectans Peck, Bull. Torr. Bot. Club ii:26. 1884.
Pileus sessile, sometimes imbricate, dimidiate in outline,
3-7 x 4-15 X 0.7-3 cm., rather spongy and watery when fresh,
firm and rigid when dry, white or whitish, finely tomentose
or glabrous, azonate, margin thin and acute; context white,
in large specimens duplex, with a firm lower layer and a soft
upper layer, in smaller specimens more uniform, 0.5-1.5 cm.
thick; tubes 0.5-1.5 cm. long, mouths white, yellowish on dry-
ing, circular to angular, large, averaging 1-2 to a mm.; spores
white, smooth, ellipsoid to globose, 4.5-5.5 x 6.5-8.5 ju.
On diseased or injured trunks of deciduous trees, especially
Acer; sometimes on logs of Fagus. September to December.
Frequent.
The species is separated from P. spumeus Sow. ex Fries
by the larger tube mouths and the less globose spores that have
r
f
^
I
n
[Vol. 1
100 ANNALS OF THE MISSOURI BOTANICAL GARDEN
not been observed to be uninucleate as in that species. It
is a large white fungus distinct from the other allied species
in size, length of tubes, and habitat.
17. P. obtusus Berk. Ann. & Mag. Nat. Hist. I. 3 : 390. 1839.
Plants annual, sessile, sometimes imbricate; pileus dimidiate,
convex or ungulate, 3-9 x 4-15 x 3-6 cm., somewhat spongy
when fresh, firm, rigid, and very light in weight when dry,
cinereous to yellowish or darker in herbarium specimens,
hirtose-tomentose, rarely becoming glabrous, azonate, margin
thick, obtuse; context white or whitish, spongy to corky, some-
times duplex, 1-3 cm. thick; tubes 1.5-3 cm. long, the mouths
white, bay or brown on drying, circular to angular and sinuous,
1 mm. or more in diameter; spores (teste Murrill) globose,
smooth, hyaline, 6-8 n.
On trunks of diseased deciduous trees, especially Quercus.
R
Always easily recognized by the rounded and obtuse margin,
and the long tubes with large mouths. Excellent illustrations
are given by Spaulding (Ann. Kept. Mo. Bot. Gard. 16 : pi. 13-19)^
18. P. guttulatus Peck, in Sacc. Syll. Fung. 6: 106. 1888.
P. maculatus Peck, Ann. Kept. N. Y. State Mus. 26: 69. 1874.
Pileus sessile, sometimes imbricate, dimidiate, 3-8 x 5-12
x 0.4-1.5 cm., soft and fleshy when fresh, firm and rigid when
dry, white to yellowish or slightly brownish, glabrous, azonate
or sometimes zonate on the margin, sometimes marked with
rounded depressed spots, margin thin, acute; context white
or palHd, soft and fleshy when fresh, soft-corky or friable when
dry, 0.4-1 cm. thick; tubes 1-5 mm. long, the mouths white
to yellowish or umbrinous, angular, averaging 4-5 to a mm.;
spores white, smooth, oblong-ellipsoid, 2.5-3 x 3-5 m. (Cf . Mur-
rill, globose, smooth, hyaline, 5 /* in diameter.)
Growing on wood of coniferous trees. Rare.
The distinguishing character of the species is the presence
of the round depressed spots on the pileus.
19. P. borealis Fries, Syst. Myc. i: 366. 1821.
Pileus sessile, dimidiate, sometimes with an attenuate base,
3-8 X 4-12 x 0.5-2.5 cm., somewhat watery and spongy when
fresh, rigid when dry, white or yellowish, sometimes brownish,
hispid to tomentose, azonate, margin thin and acute; context
1
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OVERHOLTS — THE POLYPORACEiE OF OHIO 101
white or yellowish^ distinctly duplex, firm and fibrous below,
soft and floccose above, 0.5-2 cm. thick; tubes 3-10 mm. long,
the mouths white or yellowish, angular to irregular and uneven,
rather large, averaging 2-3 to a mm.; spores (teste Murrill)
ovoid, smooth, hyahne, 5-6 x 3-4 /x.
Growing only on trunks of coniferous trees. Rare.
The species is most easily separated from its allies by the
size and habitat. For illustrations see Atkinson, Mushrooms
/. 9., Duggar, Fung. Dis. Plants/. 228., and Atkinson, Cornell
Univ. Agr. Exp. Sta. Bui. 193:/. 63.
J-
20. P. Spraguei Berk. & Curt. Grevillea i : 50. 1872.
Plants annual, sessile or decurrent, sometimes imbricate;
pileus dimidiate, 4-12 x 4-10 x 0.6-2 cm., fleshy-tough when
fresh, rigid when dry, white or cinereous, appressed-tomentose
or glabrous, azonate or somewhat zonate, margin thin or rather
thick, acute, often blackening on drying; context white, watery,
tough-fibrous when fresh, sometimes very hard when dry, zonate,
0.3-1.5 cm. thick, with a disagreeable odor in fresh specimens;
tubes 0.3-1 cm. long, mouths white or discolored, circular or
angular, averaging 3-4 to a mm.; spores (teste Murrill) ellipsoidal
smooth, hyaline, 6x4//.
On dead wood of deciduous trees, especially on Fagus, Quer-
cus, and Castanea. July to September. Common.
Fresh specimens are always easily distinguished by the very
disagreeable odor. Dried plants are characteristically very
hard and rigid, the context almost bony in texture.
21. P. zonalis Berk. Ann. & Mag. Nat. Hist. I. io:375.
1842.
effused-reflexed
pileus dimidiate or laterally confluent, 0-2.5 x 1-5 x 0.2-0.^
cm., fleshy and pliable when fresh, rigid and firm when dry
whitish to flesh-colored or isabeUine, finely tomentose to gla-
brous, at first azonate but becoming zoned when mature, th(
margin at first thick, thin with age; context white, fibrous wher
fresh, hard and rigid when dry, 1-2 mm. thick; tubes 1-3 mm
long, the mouths usually more or less flesh-tinted when fresh
angular, averaging 4-5 to a mm., the walls thick and entire
very firm and rigid on drying; spores white, smooth, globose
2.5-5 a broad, with one large nucleus.
i*'.
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102 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
On old rotting logs, especially of Liriodendr on . August to
December. Not common.
The writer has collected this plant several times in the IMiami
valley, almost always on logs of Liriodendron tuUpifera. The
plant is usually entirely resupinate and has doubtless been de-
scribed as a Poria, but good collections were made which showed
beyond a doubt the plicate tendency of the plant. No dispo-
sition could be made of the plant until Dr. Alurrill suggested that
it might belong to P. zonalis. Later, specimens were sent to
Rev. Bresadola who pronounced it that species and an opinion
recently received from Mr. Lloyd expresses the same view. It
is, however, quite different from the usual forms of that plant
and the name is used with some apprehension. The plant
is also abundant in Missouri where the writer has found the
pileate forms to be much more common than in Ohio. P,
zonalis has been supposed to be confined to the Gulf States in
this country, although it is not surprising that semi-tropical
forms found there should extend their range up the large river
valleys to the north.
22. P. adustus Willd. ex Fries, Syst. Myc. i: 363. 182L
Boletus adustus Willd. Fl. Berol. 392. 1787.
Plants annual, sessile, effused-reflexed, or resupinate; pileus
dimidiate, often imbricate, 1-6 x 2-7 x 0.2-0.4 cm., fleshy-
tough when fresh, coriaceous or rigid when dry, white to cin-
ereous or pale tan, fibrillose-tomentose to almost glabrous, zonate
or azonate, the surface usually rough, margin thick and broadly
sterile below when young, becoming thin when mature; context
white or paUid, rather soft when fresh, corky or fibrous-corky
when dry, 1-3.5 mm. thick; tubes not more than 1 mm. long,
the mouths grayish black to black, angular, even, minute,
averaging 5-7 to a mm.; spores white, smooth, oblong to oblong-
ellipsoid, 2-2.5 X 3.8-4.3 //.
On stumps and trunks of dead deciduous trees. August to
December.
This species differs from P. fumosus Pers. ex Fries and P.
fragrans Peck in the smaller size and the uniformly black
hymenium.
V
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OVERHOLTS — THE POLYPORACE^ OF OHIO 103
33. P. fragrans Peck, Rept. N. Y. State Museum 30: 45.
1879.
Plants annual, sessile or effused-reflexed; pileus dimidiate,
imbricate, 2-8 x 4-10 x 0.5-2 cm., fleshy-tough when fresh,
firm and rigid when dry, cinereous to reddish gray, finely tomen-
tose to almost glabrous, subzonate or azonate, the margin thin
and acute; context whitish or pallid, tough when fresh, soft-
corky when dry, 4-8 mm. thick, with a sweet anise-like odor
J
that persists in dried plants; hymenium sometimes separated
from the context by a narrow, dark-colored line; tubes less than
4 mm. long, the mouths whitish or somewhat smoke-colored,
blackish when bruised, angular, the dissepiments becoming
dentate and the mouths unequal in size, averaging 3-4 to a mm. ;
spores (teste Murrill) white, globose to ovoid, smooth, 5-6 /^ in
diameter.
On stumps and trunks, especially of Ulmus. Frequent.
The distinguishing characters of this species are the fragrant
odor and the unequal and irregular pores — characters which
separate it from P. adustus and P. fumosus. The name P.
puherula Berk. & Curtis is sometimes applied to this plant.
24. P. fumosus Pers. ex Fries, Syst. Myc. i : 367. 1821.
Boletus fumosus Pers. Syn. Fung. 530. 1801.
Plants annual, sessile or effused-reflexed; pileus dimidiate,
often imbricate, 2-7 x 3-8.5 x 0.3-2 cm., somewhat fleshy-
tough when fresh, firm and rigid when dry, grayish to very pale
tan-colored, finely tomentose, subzonate or azonate, margin thin
and acute; context white to light umber, soft corky when fresh,
corky when dry, 0.3-2 cm. thick, with a rather disagreeable
odor; hymenium separated from the context by a distinct,
narrow, dark-colored line; tubes short, not more than 3 mm. long,
the mouths whitish or smoky, blackish when bruised, circular
to somewhat angular but thick-walled and entire, averaging
4-6 to a mm., spores white, smooth, elliptical to subcylindrical,
2.6-4 x 5.3-7.2 m.
■
Growing on dead wood of deciduous trees. October to Decem-
ber. Frequent.
Distinguished from P. fragrans Peck by the more circular
and entire tube mouths and, in our plants at least, by the absence
of the fragrant, anise-like odor. The odor is disagreeable in
[Vol. 1
104 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the fresh plants but disappears on drying. Bresadola ascribes
a subanise odor to the plant at times. The plants are, however,
closely related and one may expect to find intermediate forms
that are difficult to refer to either species. Thin, semi-resupi-
nate forms are often scarcely distinguishable from P. adustus
Willd. ex Fries. The plant is illustrated by Bresadola (Fungi
Trident, pi 135),
2$. P. robiniophila (Murr.) Overholts, n. comb.
Trametes robiniophila Murr. N. Am. Flora 9: 42. 1907.
Plants annual, sessile, rarely imbricate; pileus dimidiate,
fleshy-tough or somewhat coriaceous when fresh, firm and rigid
when dry, 3.5-10 x 4-15 x 1-4 cm., white to cinereous or yel-
lowish, finely tomentose to glabrous, azonate or rarely sub-
zonate or concentrically sulcate in large specimens, margin at
first thick and obtuse, becoming thin and acute when mature;
context white, fleshy-tough when fresh, soft and punky when
dry, 0.5-3 cm. thick, usually with a sweet anise-like odor devel-
oping in herbarium specimens; tubes 0.3-1 cm. long, mouths
white, often bay or brownish in dried plants, circular to angular,
averaging 4-6 to a mm,, the walls thick and entire; spores
white, smooth, ovoid to subglobose, 5.5-7 x 7-8.5 n.
On deciduous trees, especially Robinia, Celiis, and Acer.
August to December. Common.
Dried plants are characterized by the tough, punky context
and the sweet odor, as well as by the large size of the plant,
the long tubes, the minute mouths, and the habitat. The plant
was first described as a Trametes but it appears to belong rather
to Polyporus.
26. P. betulinus Bull, ex Fries, Syst. Myc. i : 358. 1821.
Boletus betulinus Bull. Herb. Fr. pi 312. 1786.
Pileus sessile or attached by a prominent lateral umbo, dimidi-
ate to circular in outline, 3-9 x 3-15 x 1-5 cm., somewhat
fleshy when young, firm and rigid when dry, glabrous, azonate,
smooth, covered with a thin pellicle, margin more or less incurved,
with a wide sterile band on the lower surface; context white,
somewhat fleshy when fresh, soft-corky when dry, 1-3.5 cm.
thick: tubes 3-8 mm. loner, mouths white, circular to anffiilar.
averaging 3-4
times covered by
I
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r
sometimes as much as 2 mm. Ions:: tube
1
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OVERHOLTS — THE POLYPORACEJE OF OHIO 105
ing in a smooth layer from the context; spores (teste Murrill)
white, cylindrical, curved, 4-5 n long.
Growing only on Betula. Not common.
Always easily recognized by the habitat, the smooth, pellic-
ulose surface and the inrolled, broadly sterile margin of the pil-
eus. Good illustrations are given by Freeman (Minn. Plant
Diseases /. 1^6), Hard (Mushrooms /. 337), White (Hymen.
Conn. pi. 37), and Kellerman (Ohio Myc. Bui. 10:/. ^3).
37. P. volvatus Peck, Kept. N. Y. State Mus. 27: 98. 1875.
Plants annual, sessile or attached by a stem-like base; pileus
globose or compressed-globose in form, 1-5.5 cm. broad, 1-
3.5 cm. thick, somewhat coriaceous-corky when fresh, hard and
firm when dry, somewhat encrusted, whitish or yellowish, some-
times tinged with red, glabrous, azonate, margin thick and
rounded, extending downward and backward and forming a veil-
like covering over the hymenium; context white or hght colored,
soft-corky, 0.2-1 cm. thick; tubes 2-5 mm. long, the mouths
whitish to brownish, circular, averaging 3-4 to a mm.; the cover-
ing over the hymenium ruptures in from one to three places and
allows the escape of the spores; spores (teste Peck) flesh-
colored, elliptical, 5 x 7.5-9 ju.
On dead wood of coniferous trees. Rare.
An aberrant form easily recognized by the veil-like covering
of the hymenium. This is persistent, being coriaceous in tex-
ture and as much as 1 mm. thick. Peck's illustration (Kept.
N. Y. State Mus. 27. yl. 2. f. 3-6) gives some idea as to the
general form of the plant; Hard's (Mushrooms /. 31^0) is not
much better, von Schrenk gives a good illustration (U. S.
Dept. Agr., Div. \
/
28. P. distortus Schw. ex Fries, Elench. Fung, i : 79. 1828.
Boletus distortus Schw. Syn. Fung. Car. 97. 1822. Poly-
porus ahortivus Peck, Bot. Gaz. 6: 274. 1881.
Plants stipitate or substipitate, variable in form and size,
sometimes with a distinct, well developed, centrally placed
stipe, sometimes the whole plant distorted and the stipe rudi-
mentary, often almost the entire surface of such forms covered
with the tubes; pileus circular to irregular in outline, fleshy-
tough when fresh, firm and coriaceous when dry, variable in
color, whitish, grayish, tan-colored, rufescent, or brownish, vil-
;
mm
^f.^ [Vol. 1
lUO ANNALS OF THE MISSOURI BOTANICAL GARDEN
lous-tomentose, soft to the touch, azonate, margin thin and acute
or thick and obtuse; context white or whitish, with a firm corky
layer next to the hymenium and a lighter colored, softer layer
above, the whole 0.2-1 cm. thick; tubes in well developed speci-
mens 1-6 mm. long, whitish or rufescent when bruised, mouths
angular to da^daloid and irregular, averaging 1-3 to a
stipe central, lateral, or wanting, rarely well developed and up to
6 cm. long, more often rudimentary and tubercular, clothed
like the pileus, soft on the outside and firm within; spores
white, smooth, subglobose, 5.5-8.5 ^ in diameter; conidial (?)
spores sometimes present, white, smooth, ovoid to elliptical
3.3-4.2 X 5.2-7.8 ai.
Usually growing about stumps and probably always attached
to buried w^ood. Common.
^ Well developed specimens of this plant will be easily recog-
nized by the duplex context and the soft, villous pileus; abnor-
mal specimens by their distorted appearance. The duplex
context is always more easily recognized in dried specimens.
According to Lloyd our plant is identical with P. rufescens Fries
of Europe. See Lloyd, Syn. Stip. Polyp. /. 458., for illustration
of one form of the distorted plant.
29. P. pocula Schw. ex Berk. & Curt. Proc. Am. Acad. Arts
Sci. 4: 122. 1858.
Sphaeria pocula Schw. Jour. Acad. Nat. Sci. Phil. 5: 7. 1825.
Enslinia pocula Schw. ex Fries, Summ. Veg. Scand. 2
1849
399
^ Pileus short-stipitate, pendant from dead branches, circular
m outhne, 1-5 mm. in diameter, 1-3 mm. thick, coriaceous when
fresh, rigid when dry, whitish to brown in color, pruinose or
mealy, azonate; context coriaceous when fresh, hard when dry,
less than 1 mm. thick; tubes not more than 0.5 mm. longi
mouths at first appearing pruinose, whitish or brow
cir-
cular, very minute, averaging 5-6 to a mm.; stipe dorsally
attached, concolorous with and expanding into the pileus, prui-
nose, not more than 5 mm. long; spores (teste Murrill) globose,
smooth, hyaline, 4 /i in diameter.
On dead branches, especially of Quercus and Castanea. Rare.
This is the smallest known polypore and easily identified by
its size and habit of growth. It was first decribed as an asco-
<
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 107
mycete {Sphaeria) by Schweinitz and later transferred to the
m
genus EnsUnia (Pyrenomycetes) by Fries. Excellent illustra-
tions are given by Lloyd (Myc. Notes, Polyp. Issue 3: /. 369-
70; Syn. Stip. Polyp. /. 4^5).
30. P. brumalis Pers. ex Fries, Syst. Myc. i: 348. 1821.
Boletus hrumalis Pers. Neues Mag. Bot. i : 107, 1794.
Pileus stipitate, circular in outline, sometimes somewhat urn-
bilicate in the center, 1.5-5 cm. broad, 0.2-0.4 cm. thick,
fleshy-tough when fresh, rigid when dry, varying in color from
yellowish brown to dark brown or almost black, minutely
hispid to glabrous, rarely slightly squamulose, usually azonate
but at times distinctly zoned, margin thin and entire, involute
when young and incurved on drying; context white or pallid,
soft-fibrous when fresh, firm when dry, 2 mm. or less thick;
tubes 1-3 mm. long, usually slightly decurrent, the mouths
white or whitish, at first circular and thick walled, later angu-
lar and the dissepiments thinner, averaging 2-3 to a mm.;
stipe central or subcentral, simple, cylindrical, grayish or brown-
ish, minutely hispid or glabrous, 2-3 cm. long, 0.2-0.3 cm. thick;
spores white, oblong, sometimes slightly curved at one end,
smooth, 2.5 x 9 a*.
Growing on dead wood in the fall and early winter. Common.
P. brumalis and P. arcularius are closely related species that
are not always easy to separate. In general the forms occur-
ring in the early spring and summer are likely to be P. arcularius,
while those found in autumn and often late in winter are more
likely to be P. brumalis. Hard (Mushrooms/. 335) gives a good
illustration of the plant.
31. P. arcularius Batsch. ex Fries, Syst. Myc. i: 342. 1821.
Boletus arcularius Batsch. El. Fung. 97. 1783. P. arcular-
iformis Murrill, Torreya 4: 151. 1904.
Pileus stipitate, circular in outline, convex to umbilicate,
sometimes infundibuliform, 1-8 cm. broad, 1-4 mm. thick,
fleshy-tough or coriaceous when fresh, rigid when dry, golden
brown to dark brown, usually more or less squamulose, azonate,
the margin usually distinctly ciliate, involute on drying; con-
text white or pallid, fibrous-fleshy when fresh, compact-fibrous
when dry, less than 2 mm. thick; tubes 1-2 mm. long, often
decurrent, the mouths white, discolored on drying, angular and
/
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108 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
often radially elongate, averaging 2 to a mm. in transverse
direction and about 1 to a mm. in axial direction; stipe central
or subcentral, concolorous with the pileus, fuscous-squamulose
to glabrous above, often hispid at the base, 2-6 cm. long, 2-4
mm. thick; spores white, smooth, elliptical-cyHndrical, usually
2-3 guttulate, 2-3 x 6-8.5 ^.
On dead wood. Common.
This species is much more common than the preceding and
is distinguished from it by the hghter colored pileus, the ciliate
margin, the hispid stipe base, and the larger and more alveolar
tubes. Is is usually found in the spring and early summer. A
small form of it with the pileus not more than 1 cm. in diameter
is especially common on twigs and bits of wood during the late
spring and early summer. Murrill regards this form as a dis-
tinct species and has named it P. arculariformis (Torreya 4:
151). It is here maintained as a form of P. arcularius. This spe-
cies is w^ell represented by Hard (Mushrooms/. 336).
32. P. pennsylvanicus Sumstine, Jour. Myc. 13: 137. 1907.
Pileus stipitate, circular in outline, depressed, sometimes
umbilicate or somewhat infundibuliform, 4-6.5 cm. in diameter,
0.2-0.5 cm. thick, fleshy-tough, pale tan or ochraceous buff in
color, with a thin cuticle, glabrous, azonate, margin thin and
acute; context white, soft and watery when fresh, with a sweet
acid odor, rather fragile when dry, 2-4 mm. thick; tubes white
at first, discolored on drying, long decurrent on the stipe, 2-4
mm. long, mouths angular, thin walled, large, somewhat longer
in the radial direction, 1-2 mm. long, 0.5 to 1 mm. wide; stipe
central or excentric, whitish, glabrous, expanding above, 2-3
cm. long, 0.4-1 cm. thick; spores white, smooth, oblong-ellip-
tical or fusoid, 4.2-5.7 x 10-14 w. often onoe to spvr^ml
times
guttulate.
old logs in July and August. Frequent
The above description is drawn from notes and specimens
from two collections made at Oxford, Ohio, one in August,
1910, and the other in July, 1911. The odor of the fresh plant
is described by the author as "nitrous". The laree aneular
with Favolus canadensis Klotzsch. From the former
and
separated by the much larger spores and from the latter by
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1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 109
the well developed stipe with the decurrent tubes, the usually
umbiUcate pilous, and the friable context when dry. Possibly
it should be referred to P. Rostkowii Fr. or to P. pallidus Schulz.
& Kalchbr., both of which some regard as being small scaleless
forms of P. squamosus Huds. ex Fries. The spores agree well
with those of P. squamosus, but although it can be shown to be
related to that species, it is worthy of a distinct name.
33. P. squamosus Huds. ex Fries, Syst. Myc. i: 343. 1821.
Boletus squamosus Huds. Fl. Angl. 626. 1798. [2nd ed.]
Pileus short-stipitate or almost sessile, dimidiate to reniform
I
in outline, 6-25 cm. in diameter, 0.5-4 cm. thick, fleshy when
fresh, firm and rigid when dry, whitish to dingy yellowish or
brownish, clothed, especially toward the center, with large, ap-
pressed, brownish scales often concentrically arranged, azonate,
margin thin and acute; context white, tough, soft-corky when
dry, 0.5-3.5 cm. thick; tubes 2-8 mm. long, decurrent, the
mouths white or yellowish, large and angular, 1-2.5 mm. in
diameter; stipe lateral, often rudimentary, black at the base,
reticulate above by the decurrent pores, 1-5 cm. long, 1 cm. or
more thick.
Growing on injured or diseased deciduous trees. Rare.
Lloyd gives the spores as "oblong, 5-6 x 12-15 /i, hyaline,
smooth." Easily recognized by the large pores and the large,
appressed, brownish scales. The plant is well illustrated by
Bresadola (Fung. Trident, pi 133), Freeman (Minn. PL Diseas.
125), Lloyd (Photograph, pi. 5), and Hard (Mushrooms /.
325).
34. P. picipes Fries, Syst. Myc. i: 353. 1821.
P. Jissus Berk. Hooker's Lond. Jour. Bot. 6:318. 1847.
Pileus stipitate, circular to reniform in outline, convex or
plane, when older usually becoming depressed or somewhat
infundibuliform, 4-20 cm. broad, 0.1-0.8 cm. thick, tough and
leathery when fresh, very rigid and brittle when dry, sometimes
yellowish brown but usually dark chestnut-brown to reddish
brown, usually lighter in color towards the margin, azonate,
margin verv thin, usually wavy and often lobed ; context white
somewhat
when fresh, firm when dry
1-7 mm. thick: tubes not more than 2 mm. long, decurrent
{
[Vol. 1
110 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the stipe, the mouths white to brownish in color, circular to
angular, very minute, invisible to the unaided eye, averaging
5-7 to a mm.; stipe central to lateral, dark brown or black on
the lower half, glabrous, 1-6 cm. long, 0.4-1.5 cm. thick.
On stumps and logs late in autumn. Common.
The combination of black stipe base and minute pores char-
acterizes this and the next species. The two are separated
mainly on point of size. Murrill describes this plant under the
name of P. fissus Berk., which was originally described from
specimens collected in Ohio. Patouillard (Tab. Fung. No.
136) says the spores are ovoid. Lloyd now considers this plant
to be a form of P. varius Fries, of Europe. A good illustration
of our plant will be found in Hard, Mushrooms /. 319.
35. P. elegans Bull, ex Fries, Epicr. Syst. Myc. 440. 1838,
Boletus elegans Bull. Herb. Fr. jpl. 46. 1780.
Pileus stipitate, circular to reniform in outline, convexo-
ane or depressed, 1.5-7 cm. in diameter, 0.2-1 cm. thick,
leathery when fresh, rigid and firm when dry, pale ochraceous
to dull orange-color, pruinose to glabrous, azonate, the margin
rather thin, often radiate-striate, even or undulate ; context white
to light ochraceous, tough when fresh, soft corky when dry, 1-6
mm. thick; tubes 1-3 mm. long, decurrent on the stipe, the
mouth whitish to pallid, circular to angular, averaging 4-5 to a
mm.; stipe central, excentric or lateral, slender, black at the
base, light colored above, pruinose or glabrous, 1-8 cm. long,
0.2-0.6 cm. thick.
On dead wood late in autumn. Not common.
Spores were not obtained from the writer's specimens. Mur-
rill gives them as ''oblong, smooth, hyahne, 7-8 x 3-3. 5;u."
The species is closely related to P. picipes Fries but is separated
from it by the smaller size and the uniform ochraceous color
of the pileus that never takes on the darker chestnut shades
assumed by P. picipes. Bulliard (Herb. Fr. pi. 124) gives an
excellent illustration of the plant under the
name
Bull
36. P. radicatus Schw. Trans. Am. Phil. Soc. II. 4: 155.
1832.
P. Morgani Peck, Ann. Kept. N. Y. State Mus. 32: 34. 1879.
Pileus stipitate, circular in outhne, 3.5-20 cm. broad, 0.3-0.8
!
/
[
€
}
(
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1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 111
cm. thick, fleshy or fleshy-tough when fresh, more or less friable
when dry, yellowish brown or darker, finely tomentose or fibril-
lose-scaly, often becoming glabrous, azonate; margin thin and
acute, often involute on drying; context white or light yellow-
ish, soft and spongy, 2-6 mm. thick; tubes 1-5 mm. long,
decurrent on the stipe, the mouths white or brownish on drying,
circular to angular and irregular, averaging 2-3 to a mm. ; stipe
central, simple or rarely branching once or twice, yellowish or
brownish, prolonged below into a long, black, rooting base,
velvety or rough-squamulose above, 6-15 cm. long, 0.5 to 2 cm.
thick; spores white, smooth, ovoid-elliptical, 6-8 x 12-15 ix.
Growing on the ground, sometimes around stumps, and prob-
ably attached to buried wood. Common.
This plant is always easily recognized by the black and radi-
cating base of the stem. The type specimens of P. Morgani
Peck were collected in Ohio by Morgan. For illustrations see
Hard, Mushrooms /. 329., Lloyd, Syn. Sec. Ovinus /. 508;
Syn. Stip. Polyp./. J^65., and Ohio Myc. Bull. 11:/. 4^.
37. P. flavovirens Berk. & Curt. Grev. i : 38. 1872.
Pileus stipitate, circular to irregular in outline, 4-10 cm.
broad, 0.3-0.8 cm. thick, soft and fleshy when fresh, rigid but
friable when dry, yellowish green or yellowish brown in color,
h ^^ ^^
the surface often cracked and areolate and the flesh showing
yellowish in the cracks, slightly tomentose or glabrous, azonate,
the margin thin and acute ; context white or yellow, fleshy when
fresh, soft and friable when dry, 1-4 mm. thick; tubes 1-5 mm.
long, decurrent on the stipe, the mouths white or yellowish,
sometimes reddish on drying, circular to angular, averaging 1-3
to a mm. ; stipe simple or branched, usually excentric but some-
times central, often irregular in form, whitish or yellowish in
color, 3-6 cm. long, 1-1.5 cm. thick; spores white, smooth,
globose, or subglobose, 3-4.7 n in diameter.
Growing on the ground in deciduous woods. Frequent in
July and August.
A species easily recognized by the color of the pileus. The
plant is fairly well represented by Hard (Mushrooms /. 327),
and by Lloyd (Syn. Sect. Ovinus/. 501). According to Lloyd
P, cristatus Pers. of Europe is not different from our plant.
Murrill lists it under the name of Grifola poripes Fries ex Murr.
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112 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
38. P. umbellatus Pers. ex Fries, Syst. Myc. i:354. 1821.
Boletus umbellatus Pers. Syn. Fung. 519. 1801.
Plants stipitate, 7-20 cm. in diameter, the stipe branching
repeatedly and giving rise to many centrally attached pileoli
which are circular in outline, 1-4 cm. broad, less than 5 mm.
thick, fleshy in texture when fresh, rigid when dry, whitish
to smoky brown in color, fibrillose or glabrous, azonate; mar-
gin thin, acute, entire; context white, fleshy or fleshy-tough,
rather brittle when dry, usually not more than 1 mm. thick;
tubes less than 2 mm. long, decurrent on the stipe branches, the
mouths white, angular, averaging 2-4 to a mm.; stipe compound^
the branches cylindrical in form, central or subcentral, white,
usually entirely covered with the decurrent tubes; spores
white, oblong-elliptic, smooth, 2.3-3.5 x 7-9.4 /i.
Growing about the bases of stumps or trees, especially of
Quercus. Rare.
Easily distinguished from its alhes by the more regular and
cylindrical stipe branches, the small and centrally attached
r
pilei which are more or less circular in outline, and by the ob-
long-elliptic spores. Murrill describes it as Grifola ramosissima
Scop, ex Murr. The plant is well illustrated by Lloyd (Syn.
Stip. Polyp. /. 450), Hard (Mushrooms /. 320), and Atkinson
(Mushrooms/. 183).
39. P. frondosus Dicks, ex Fries, Syst. Myc, i:355. 1821.
Boletus frondosus Dickson, Fasc. PL Crypt. Brit, i : 18. 1785.
Plant stipitate, the stipe many times branching and giving
rise to. numerous overlapping pileoli, the whole plant forming
a more or less globose mass often as much as 40 cm. in diam-
eter; pileoli flabelliform or spathulate in outline, 2-7 cm, broad,
2-5 mm. thick, fleshy-tough when fresh, rigid when dry, grayish
to mouse-colored, glabrous or minutely tomentose, azonate, the
margin thin and acute; context white or whitish, fleshy-tough
when fresh, fragile when dry, not more than 2 mm. thick; tubes
2-3 mm. long, decurrent on the stipe, the mouths white, angular
or irregular, averaging 1-3 to a mm.; stipe compound, short
and thick; spores white, smooth, ovoid to elliptical, 4.5-6 x
6-9 n.
Usually found at the bases of trees or stumps, preferably
of Quercus and Ulmus. Common in late fall.
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OVERHOLTS — THE POLYPORACE^ OF OHIO 113
From p. Berkeleyi Fries and P. giganteus Fries this species
is separated by the numerous small pileoli which in those species
are large and few in number. The irregular stipe-branches and
the more spathulate pileoli separate it from P. umhellaius
Fries in which the stipe branches are cylindrical and the pileoli
centrally attached and consequently more nearly circular in
outline. The plant is illustrated in Atkinson, Mushrooms
181-82., Hard, Mushrooms /. 321., and Mcllvaine, Am.
Fungi j)l. 128.
40. P. giganteus Pers. ex Fries, Syst. Myc. i : 356. 1821.
Boletus giganteus Pers. Syn. Fung. 521. 1801. Grifola Sums-
tinei Murrill, Bull. Torr. Club 31: 335. 1904.
Plants composed of a few broad pileoli, 6-15 cm. in diameter
and less than 0.5 cm. thick, dimidiate to flabelliform or spathu-
late in outline, fleshy-fibrous when fresh, more rigid when dry,
grayish to brown, often black when dried — especially on the
margin — ,usually somewhat tomentose or fibrillose, azonate or
subzonate, margin very thin and acute, often lobcd, involute on
drying; context white, fibrous, tough, 1-3 mm. thick; tubes
1-3 mm. long, at first white but blackish where bruised and
on drying, the mouth angular to irregular, often torn, averaging
5-7 to a mm.; stipe short and thick; spores white, smooth,
globose, 4-6 m broad.
Growing on the ground around stumps. Frequent.
Separated from P. Berkeleyi Fries by the smooth spores;
from P. umbellatus Pers. ex Fries, and P. frondosus Fries, by
the much larger and fewer pileoli, and distinct from all of these
in the blackening of the margin or of the entire pileus and
hymenium when bruised or in drjdng. In the 'North American
Flora' it is described under the name of Grifola Sumstinei Murr.
In this country it has always been held to be the same as the
European plant P. giganteus Fries, and European specimens
recently received from Bresadola confirm this view. A very
good illustration will be found in Bresadola, Fungi Tridenti
pi. 134., and in Boudier, Ic. Myc. i: pi. 163. To the writer's
knowledge it has not been illustrated in American mycology.
41. P. Berkeleyi Fries, Nov. Sym. 40. 1851.
P. anax Berk. Grev. 12: 37. 1882.
1
Pileus stipitate, the stipe sometimes branching and giving
[Vol. 1
114 ANNALS OF THE MISSOURI BOTANICAL GARDEN
more
) from 2 to 4 pileoli, sometimes simple with but one large
; pileoli fleshy-tough when fresh, becoming rigid on drying,
or less circular in outline, 6-15 cm. broad, 0.3-1.5 cm.
thick, hght colored, whitish to yellowish, sHghtly tomentose
or glabrous, azonate or obscurely zoned; margin rather thin,
often lobed; context white, fleshy-tough, fragile when dry,
0.3-2 cm. thick; tubes 2-8 mm. long, decurrent on the stipe;
mouths white or whitish, large and irregular, averaging 0.5-2
mm. in diameter; stipe short and thick, more or less tubercular,
whitish in color, 4-7 cm. long, 3-5 cm. thick; spores white,
minutely echinulate, globose, 5.6-8.4 n in diameter.
Growing at the bases of trees and stumps, especially of Quercus.
Frequent.
This is one of the largest of our species and is easily distin-
guished from all of its allies by the echinulate spores. Morgan's
description of P. anax Berk, applies to P. frondosus Fries and
not to P. Berkeleyi for which P. anax is a synonym. (See
Lloyd, Mycological Notes 27: 341-342.) The plant is well
represented by the following illustrations: Lloyd, Photogr.
pi 9-10; Myc. Notes Polyp. Iss. 3: /. 362-63., and Hard,
Mushrooms /. 323 and pi. 1^5.
42. P. sulphureus Bull, ex Fries, Syst. Myc. i: 357. 1821.
Boletus sulphureus Bull. Herb. Fr. pi. 4^9. 1788. P.
cincinnatus Morgan, Jour. Cine. Soc. Nat. Hist. 8: 97. 1885.
Plants annual, often attenuate at the base and appearing
substipitate, imbricate; pileus dimidiate to flabelliform in
outUne, 5-20 x 4-12 x 0.5-2.5 cm., fleshy and watery when
young, becoming firm when old, yellowish to bright orange-
colored, sometimes fading with age, finely tomentose to gla-
brous, azonate or with broad colored zones, the margin thin
and acute, sometimes lobed; context white or light yellow,
fleshy when fresh, rather soft and friable when dry, 0.4-2 cm.
thick; tubes 1-4 mm. long, the mouths bright sulphur-yellow,
sometimes whitish or dull yellow with age or on drying, angular,
averaging 2-4^ to a mm.; spores white, smooth, ovoid to sub-
globose, 4-5 x 5.5-7 m-
Growing on trunks and stumps of deciduous trees. Common.
Specimens usually change color on drying and most of the
red color of the pileus is lost. The bright yellow of the hyme-
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OVERHOLTS — THE POLYPORACEiE OF OHIO 115
nium may or may not persist. The best colored representation
of the fungus is that given by Rostkowius in Sturm, Deutschl.
Flora 4 : pL 20. The plant is widely distributed and well known
and has figured largely in American mycology. The following
illustrations will aid in determinations: Atkinson, Mushrooms
/. 184-85,, Duggar, Fung. Dis. PL /. 226. y Hard, Mushrooms
/. 326.y and von Schrenk, U. S. Dept. Agr., Div. Veg. Path. Bui.
2b: pi. 11. f. 1-4.
43. P. Pilotae Schw. Trans. Am. Phil. Soc. II. 4: 157.
1832.
P. hypococcineus Berk, Lond. Jour. Bot. 6:319. 1847.
Plants annual, sessile; pileus dimidiate, often subungulate,
5-12 X 6-15 X 1-5 cm., soft coriaceous or corky, buff or orange-
]
colored, becoming whitish on drying, minutely tomentose or
glabrous, azonate, margin usually obtuse; context pale buff,
becoming carneous when dry, fibrous, sometimes very hard
when dry, strongly zonate, 0.7-2 cm. thick; tubes 0.5-2 cm.
long, the mouths orange-colored, becoming brownish on drying,
angular, averaging 3-5 to a mm. ; spores (teste Murrill) smooth,
hyaline, 3-4 x 2-3 a*.
On dead wood of Quercus and Castanea. Rare.
Easily distinguished from other species with a predominance
of red or orange colors by the thick pileus and the long tubes.
The plant is said to emit a strong odor when growing. The
type specimens of P. hypococcineus Berk, were collected in
Ohio by Lea. P. castanophilus Atk., described from North
Carolina, is said to be the same plant.
44. P. sanguineus L. ex Fries, Syst. Myc. i: 371. 1821.
Boletus sanguineus L. Sp. Plant. 1646. 1762. [2nd ed.]
Plants annual, sessile; pileus dimidiate to flabelliform,
2-5 X 2-8 x 0.2-0.5 cm., coriaceous, bright red, finely tomentose
to glabrous, often zonate, the margin very thin and acute;
context red or yellowish red, soft and floccose, scarcely more
than 2 mm. thick; tubes 0.5-1.5 mm. long, the mouths red,
more or less angular or circular when young, averaging 2-4
to a mm. ; pileus often attached by an attenuate base and then
F
appearing subs tipit ate. ,
On dead wood of deciduous trees. September to December.
I
Rare.
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[Vol. 1
116 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
The species is distinguished from the following one by the
much thinner pileus and the marked tendency to appear sub-
etipitate. OtherTvdse it scarcely differs, and intermediate forms
are found that are difficult to place satisfactorily It is usually
considered to be a southern species, but Hard reports finding it
in Ohio. His specimens were determined by Peck.
h
45. P. cinnabarinus Jacq. ex Fries, Syst. Myc. i : 371. 1821.
Boletus cinnabarinus Jacq. Fl. Austr. 4:2. 1776.
Plants annual, rarely reviving, sessile or effused-reflexed ;
pileus dimidiate or reniform, 2-6 x 2-10 x 0.5-2 cm., tough
and leathery when fresh, more rigid when dry, orange-colored
to cinnabar-red, often becoming paler or almost white with age,
compactly tomentose or glabrous, usually azonate, margin
thin or thick, acute; context red or yellowish red, floccose-
fibrous to soft-corky, always zonate, 0.4-1.5 cm. thick; tubes
1-4 mm. long, the mouths cinnabar-red, circular then angular
and sometimes somewhat sinuous, averaging 2-4 to a mm.;
spores white, smooth, oblong, 2-2.5 x 4.5-5.5 m-
On dead wood of all kinds. September to December. Com-
mon.
The prevailing deep red color of both pileus and hymenium
separates this species from all others of the genus except P.
sanguineus Fries, from which this species differs only in being
thicker and in having the context more strongly zoned. P.
cinnabarinus is a northern species and much more common in
Ohio than is P. sanguineus.
46. P. resinosus Schrad. ex Fries, Syst. Myc. i: 361. 1821.
Boletus resinosus Schrad. Spic. Fl. Ger. 171. 1794.
Plants annual, sessile or decurrent, more or less imbricate;
pileus dimidiate, 5-15 x 7-25 x 0.8-2.5 cm., somewhat fleshy
and full of water when young, firmer when mature and soft-
corky on drying, velvety-tomentose to glabrous, sulcate or with
a few broad, colored zones, margin at jBrst thick and somewhat
obtuse, becoming thinner and acute; context pallid to light
brown, fleshy and watery when young, soft-corky when dry,
0.5-2 cm. thick; tubes 1-6 mm. long, the mouths white to
pallid, changing to a darker color on drying, circular to angular,
averaging 4-6 to a mm.; spores white, smooth, cylindrical,
curved, 1.2-2 x 5-6.3 m-
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OVERHOLTS — THE POLYPORACE^E OF OHIO 117
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On old logs and stumps in October and November. Common.
Distinguished by the brown pileus and the light brown,
almost whitish context. For illustration see Hard, Mush-
rooms /. S31.
47. P. nidulans Fries, Syst. Myc. i:362. 1821.
Plants sessile or effused-reflexed; pileus dimidiate, 1.5-6 x
2-8 X 0.5-2 cm., very soft, spongy, and full of water when
fresh, firm and friable when dry, umber to cinnamon or tawny
brown, finely villous-tomentose to glabrous, azonate, margin
thin and acute, purpUsh or reddish where bruised; context con-
colorous with the pileus, sometimes with a darker layer next
to the hymenium, soft and watery when fresh, cheesy and
friable when dry, 2-8 mm. thick; tubes 2-7 mm. long, mouths
hoary when young, yellowish or reddish brown when mature,
angular or sinuous, averaging 3-4 to a mm.; spores white,
smooth, globose or subglobose, 2-3.5 m in diameter.
On dead wood of deciduous trees, especially Quercus. June
w
to September. Not common.
Distinguished by the uniform umber brown color of the
whole plant, the soft and watery context, etc.
48. P. gilvus Schw. ex Fries, Elench. Fung, i: 104. 1828.
Boletus gilvus Schw. Syn. Fung. Car. 96. 1822.
Plants annual or reviving for two or three years, sessile or
effused-reflexed, often imbricate; pileus dimidiate, 1-7 x 2-12 x
0.2-2 cm., leathery or corky when fresh, woody and rigid when
dry, yellowish brown or reddish brown, in very young stages
covered by a purplish, villous pubescence, otherwise glabrous,
usually rough, more or less zonate, margin thin and acute;
context yellowish brown, soft-corky to woody, 0.1-1.3 cm.
thick; tubes 1-5 mm. long, the mouths reddish brown or darker,
circular, then angular, averaging 6-8 to a mm., the walls rather
thick and entire; spores white, smooth, oblong-ellipsoid, 3-4
X 5-6 At.
w
On dead wood of all kinds. July to December. Common.
Closely related to P. radiatus Sow. ex Fries and P. cuticularis
Bull, ex Fries, but distinct in the white spores, the lighter
colored surface and the more woody context. P. isidiodes
Schw. as reported by Lea belongs here.
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118 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
49. P. radiatus Sow. ex Fries, Syst. Myc. i: 369. 1821.
Boletus radiatus Sow. Eng. Fungi pi. 196. 1799.
Plants annual, sessile or decurrent; pileus dimidiate or flabel-
liform and attached by an attenuate base, 2-5 x 2-7 x 0.3-2
cm., firm and rigid, yellowish brown or rust-colored, velvety
to glabrous, sometimes conspicuously zonate, sometimes azonate,
margin thin or thick, acute; context yellowish to rusty brown,
corky and somewhat friable, 2-5 mm. thick; tubes 1-8 mm.
long, the mouths grayish umber to rusty red, circular, then
angular, averaging 4-5 to a mm.; spores (teste Bresadola)
yellowish, elliptical, 3.5-4.5 x 5.5-6.5 m.
Growing commonly on Betula and Alnus. Rare.
A species intermediate between P. gilvus Schw. ex Fries,
and P. cuticularis Bull, ex Fries, distinguished from the former
by the habitat, the brighter color and the smoother surface
of the pileus, and by the colored spores, and from the latter
chiefly in the habitat. The species was reported from Ohio by
Lea but I have not examined the plants.
56. P. cuticularis Bull, ex Fries, Syst. Myc. i : 363. 1821.
Boletus cuticularis Bull. Herb. Fr. pi 462. 1809.
Plants annual, sessile, often imbricate; pileus dimidiate or
flabelliform and attached by an attenuate base, 3-7 x 3.5-10
X 0.3-1 cm., spongy and fleshy-tough when fresh, leathery to
rigid when dry, yellowish brown to rusty brown, compact
wooly-tomentose, becoming fibrillose or almost glabrous, some-
times subzonate on the margin, margin thin, acute, often
inflexed; context yellowish brown or rust-colored, tough and
watery when fresh, distinctly fibrous, 2-7 mm. thick; tubes
2-7 mm. long, the mouths hoary brown to rust-colored, angular,
averaging 3-5 to a mm.; spores yellowish brown, smooth,
mibglobose to broadly elliptical, 4.2-5.7 x 5.5-7 ju.
On dead wood of deciduous trees. August to November.
Common.
This species is very closely related to P. radiatus Sow. ex
Fries, but Ohio plants may be distinguished from that species
by the habitat, the thicker and larger pileus, and by the more
tomentose and spongy surface. P. perplexus Peck, the types
of which have been destroyed, is thought by some to be this
species and our plants are frequently referred to it.
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OVEKHOLTS — THE POLYPORACE^ OF OHIO 119
51. P. hispidus Bull, ex Fries, Syst. Myc. i:362. 1821.
Boletus hispidus Bull. Herb. Fr. pi 210. 1791. Polyporus
endocrocinus Berk. Hooker's Lond. Jour. Bot. 6:320. 1847.
Plants annual, sessile, sometimes imbricate; pileus dimidiate,
6-20 X 9-25 X 2-6 cm., spongy and watery when fresh, firm
and rigid when dry, yellowish brown to rusty red, soft from the
covering of the dense hirsute or hispid tomentum or pubescence,
azonate, margin thick or thin, obtuse or acute; context usually
light yellowish brown above and dark reddish brown next
to the hymenium, fibrous, firm when dry, 1-5 cm. thick; tubes
0.6-1.5 cm. long, mouths yellowish brown becoming darker
where bruised, curcular, then angular, averaging 2-4 to a mm.;
spores yellowish brown, smooth, broadly ovoid to ellipsoid,
6.5-7 X 7-9.5 ti.
On living trunks of deciduous trees. September to December.
Rare.
Much larger than P. cuticularis Bull, ex Fries, and P. radiatus
Sow. ex Fries, and expecially distinct by the hirsute or hispid
pubescence. In point of size it more nearly approaches P.
dryadeus Pers. ex Fries, and P, dryophilus Berk., but easily dis-
tinguished from them by the pubescence.
52. P. dryadeus Pers. ex Fries, Syst. Myc. i: 374. 1821.
Boletus dryadeus Pers. Obs. Myc. 3. 1799.
Plants sessile; pileus dimidiate, applanate, 6-30 x 8-35 x 2-6
cm., spongy and watery when fresh, more or less corky or
woody when dry, grayish brown to dark brown or black in
old specimens, glabrous, azonate, margin thick and obtuse,
distilling drops of water when young and growing; context
umber-brown to rust-colored, subshining when dry, soft and
watery, corky or woody on drying, 1.5-4 cm. thick; tubes
0.3-2 cm. long, mouths grayish brown, darker on drying, circular,
then angular, averaging 3-5 to a mm.; spores (teste Bresadola)
globose or subangular, smooth, yellowish, 8-9 x 7-8 /x.
On Hving trunks of Quercus. September to November.
Rare.
Very closely related to P. dryophilus Berk., and probably the
two have been confused in this country. P. dryadeus is usually
considered to be a more applanate form and much larger than
P. dryophilus. There is also said to be a decided difference
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120 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
in spore color in the two plants, P. dryadeus having much paler
spores than P. dryophilus, but for this I cannot vouch. So
far as known, P. dryadeus has not been collected in Ohio
but the species has been reported from Michigan and Kentucky.
Lloyd (Myc. Notes 36. /. 383) gives an illustration.
53. P. dryophilus Berk. Hooker's Lond. Jour. Bot. 6:321.
1847.
Plants annual, sessile; pileus dimidiate, often ungulate,
3-12 X 7-20 X 1-10 cm., rather rigid, grayish brown, to reddish
brown, scabrous with an innate, ferruginous pubescence; azo-
nate or subzonate, margin thick and obtuse; context cinnamon
or rusty brown, subshining, corky to hard and woody; tubes
0.3-2.5 cm. long, ferruginous-yellow within, the mouths cin-
namon-brown, angular, averaging 2-3 to a mm.; spores ferru-
ginous, smooth, ellipsoid to subglobose, 5 x 6.5 n.
On living Quercus and on logs. August to November. Rare.
This species was originally described from specimens collected
at Waynesville, Ohio, by Lea. To the description as given in
Lea's catalogue the following note was added: "Nearly alHed
to Polyporus dryadeus, but a smaller, more rigid species with
larger, differently colored pores. It has also much resemblance
to P. gilvus."
54- P. Schweinitzii Fries, Syst. Myc. i:351. 1821.
Plants stipitate or sessile; pileus circular to dimidiate, 5-15
cm. broad, 0.5-1.5 cm. thick, spongy to soft-corky when fresh,
firm, rigid, and sometimes friable when dry, ochraceous to
orange-colored or brown in mature specimens, strigose-tomen-
tose to almost glabrous, usually more or less zonate, margin
thin or thick, acute; context yellowish to reddish brown,
spongy when fresh, usually friable when dry, 0.2-1 cm. thick;
tubes 1-6 mm. long, the mouths yellowish, darker when bruised
and sometimes dark brown on drying, circular to angular and
soon irregular, averaging 1-3 to a mm.; stipe present and well
developed or entirely absent, central or excentric, agreeing in
color, pubescence and consistency with the pileus, 0-6 cm.
long, 1-2 cm. thick; spores (teste Lloyd) white, elliptical,
smooth, 4 x 6 /x.
Growing on or about Pinus. Autumn. Rare.
This species is a very variable one, yet quite distinct in habi-
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OVERHOLTS — THE POLYPORACEiE OF OHIO 121
tat, consistency, pubescence, color, etc. It is known in Ohio
only from a collection made at Cincinnati (now in the Lloyd
Museum) by Mr. Wm. Holden. For illustrations see Lloyd,
Myc. Notes, Polyp. Issue 1: /. 208., aud von Schrenk, U. S.
Dept. Agr., Div. Veg. Path. Bui. 25: pZ. 1. f. 1., pi 2.
55. P. circinatus Fries, Monogr. Hymen. Suec. 2 : 268. 1863.
Pileus stipitate or substipitate, circular to spathulate or
flabelliform, convex to depressed, 3-9 cm. broad, 0.3-1 cm.
thick, rather soft when fresh, firm when dry, yellowish to
umber-brown, tomentose to velvety, azonate or subzonate,
margin rather thin, acute; context yellowish to cinnamon-
brown, duplex, soft and spongy above, firm next to the tubes,
1-6 mm. thick; tubes 1.5-4 mm. long, the mouths whitish to
cinnamon, subcircular to angular, averaging 2-4 to a mni.;
stipe sometimes rudimentary, usually lateral or excentric,
fulvous to dark brown, tomentose, soft, up to 5 cm. long,
0.5-1.5 cm. thick; spores (teste Lloyd) pale color, 3 x 5 /x.
In coniferous and deciduous woods.
The species has not been reported from Ohio. It is distin-
guished by the duplex character of the context and by the poor
development of a stipe. It is a question whether it is distinct
from P. tomentosus Fr. Certainly P. dualis Peck is the same
plant. Lloyd regards American plants in which the context
is always duplex as belonging under P. circinatus Fries, and
European plants with a uniform context as P. tomentosus Fries.
The plant is illustrated by Lloyd (Myc. Notes Polyp. Issue
/
56. P. obesus (Ellis & Ev.) Overholts, n. comb.
Polystictus ohesus Ellis & Ev. Bull. Torr. Bot. Club 24:125.
1897. •
Stipitate. Stipe central, spongy, velutinous, dark cmnamon,
4-G cm. high, 0.5-1.5 cm. thick above, enlarged below to 1-3
cm.; pileus convex then depressed in the center, obconical
at first with the margin obtuse, then spreading out with the
margin acute, color lighter than that of the stipe, yellomsh
cinnamon, surface uneven, subcolliculose, not zonate, 4-6 cm.
across; pores irregular, short (1 mm.), at first round with margins
thick, finally irregular and subsinuous, 0.5-1 mm. across,
mareins acute ; spores elliptical, ferruginous, 7-8 x 4-5 ft.
[Vol. 1
122 ANNALS OF THE MISSOURI BOTANICAL GARDEN
mm
On the ground, in contact with and partly attached to decay-
ing pine limbs partly buried in the soil. (The above description
is according to Ellis and Everhardt, Bull. Torr. Bot. Club. 24:
125. 1897.)
Distinguished from the next three species by the greater
thickness of the pileus and stipe. From P. circinatus Fries,
it is separated by the absence of a duplex context and by the
slightly smaller pores. The plant is recorded by Morgan as
P. Montagnei Fries, but according to Lloyd the record is based
on plants collected in Canada by Dearness. It is listed in Lea's
catalogue under the same name.
57. P- focicola Berk. & Curt. Jour. Linn. Soc. Bot. 10:
305. 1868.
Pileus stipitate, circular in outline, convex-depressed to
umbilicate, 2-4 cm. broad, 1-6 mm. thick, coriaceous when
fresh, rigid when dry, grayish brown to cinnamon, finely tomen-
tose, striate, zonate, margin thin and acute; context cinnamon-
brown, fibrous, less than 0.5 mm. thick: tubes 1-6
the mouths angular or irregular, cinnamon to rusty brown,
averaging 1 mm. or more in diameter; stipe central, light to
dark brown, minutely velvety, 1.5-3 cm. long, 2-4 mm. thick;
spores (teste Lloyd) pale colored, smooth, elliptical, 5 x 10 /x.
On burned earth in woods. July to November. Rare.
The species differs from P. perennis L. ex Fries only in the
much larger pores. The plants were reported by Lea as P.
connatus Schw. and by Morgan as P. parvulus Klotzsch. The
plant is well illustrated by Lloyd (Myc. Notes Polyp. Issue 1:
f. 203-4).
r
58. P. perennis L. ex Fries, Syst. Myc. i:350. 1821.
Boletus perennis L. Sp. Plant. 1177. 1753.
Pileus stipitate, cu-cular in outhne, convex-depressed to
umbilicate, 1.5-7 cm. broad, 1-3 mm. thick, coriaceous, rigid
when dry, grayish brown to cinnamon or rust-colored but never
silky, finely tomentose, zonate, margin thin and acute; context
cinnamon-brown, fibrous, less than 1 mm. thick; tubes 1-2.5
mm. long, the mouths grayish to cinnamon, angular, averaging
2-4 to a mm.; stipe central or subcentral, cylindrical, con-
colorous with the pileus, velvety, 1.5-5 cm. long, 1-6 mm.
thick; spores (teste Lloyd) pale colored, 4-5 x 8-10 u.
t
1914]
OVERHOLTS — THE POLYPORACEiB OF OHIO 123
Growing on burned earth. July to November. Not common.
The plant closely resembles the next species but is separated
from it by the habitat and the dull cinnamon or cinnamon-gray
color of the zonate pileus. Polystictus proliferus Lloyd is said
by its author to be a form of this species. It was collected
near Cleveland. This species is illustrated by Atkinson (Mush-
rooms /. 187), Hard (Mushrooms /. 346), and Lloyd (Myc.
Notes Polyp. Issue 1:/. 201).
59. P. cinnamomeus Jacq. ex. Fries, Epicr. Syst. Myc. 429.
1838.
Boletus cinnamomeus Jacq. Coll. Bot. etc. i: 116. 1786.
P. suhsericeus Peck, Ann. Rept. N. Y. State Mus. 33 : 37. 1880.
Pileus stipitate, circular in outline, convex-depressed to
umbilicate, 1-5 cm. broad, 1-3 mm. thick, pliant and tough,
bright cinnamon-rufous to bright amber-brown, silky fibrillose,
the fibrils sometimes suberect towards the center of the pileus,
silky striate, sometimes zonate, margin thin and acute; context
cinnamon or rusty brown, fibrous, less than 0.5 mm. thick;
tubes not more than 2 mm. long, the mouths rufous-cinnamon,
angular, averaging 2-4 to a mm.; stipe central, cylindrical,
concolorous with the pileus, velvety to villous, 1-4 cm. long,
1-3 mm. thick; spores (teste Lloyd) pale colored, elliptical,
smooth, 5-6 x 7-10 /x.
Most frequently on clay banks, usually among moss. July
to September. Not common.
Distinguished from P. cirdnatus Fries, and P. obesus Ellis
& Ev. by the very thin context; from P. perennis L. ex Fries
by the silky pileus and the habitat; from P. focicola Berk. &
Curt, by the much smaller pores. For illustrations see Lloyd,
Myc. Notes Polyp. Issue 1 : /. 200., and Bresadola, Fungi Trid.
pi 99.
m
60. P. lucidus Leyss. ex Fries, Syst. Myc. i : 353. 1821.
Boletus lucidus Leyss. Flora Halensis 300. 1783. [2nd
ed.] Ganoderma sessile Murr. Bull. Torr. Bot. Club 29: 604.
1902. Ganoderma suhperforatum Atk. Bot. Gaz. 46: 337. 1908.
Plants stipitate or sessile, annual; pileus dimidiate or reni-
form in outUne, 3-12 x 3.5-20 x 0.4-2.5 cm., coriaceous-corky
when fresh, corky or woody when dry, the upper surface covered
by an encrusting persistent layer of deep reddish chestnut varn-
124 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
isli, often wrinkled, glabrous or pruinose from a coating of
brown conidial (?) spores, zonate or concentrically sulcate, the
margin thin and acute, sometimes lobed; context whitish to
light brown, sometimes separated into an upper, light colored,
soft layer, and a lower darker and firmer layer, but often uniform
in color and texture, 0.2-1.5 cm. thick; tubes 0.3-1.5 cm. long,
not decurrent, the mouths white or umber, darker when bruised,
circular to angular, averaging 3-5 to a mm. ; the hymenium often
with red-varnished patches on which no tubes are produced;
stipe often entirely absent, lateral when present, covered like
the pileus, 1-10 cm. long, 0.5-1 cm. thick; spores yellowish
brown, smooth or apparently slightly verrucose, ovoid with a
truncate base, 5-6.3 x 9.4-1 1;^.
On stumps and trunks of dead or injured deciduous trees.
Common.
The variation in the pileus from stipitate to sessile may be
confusing at first, but the deep chestnut-red color, not changing
to yellowish as in the next species, will usually be found to be
the distinguishing character of the species. The plant is de-
scribed by Murrill under the name of Ganoderma sessile Mutt.
Atkinson has described a new species of Ganoderma from Ohio
under the name of G. subperforatum. At the writer's request
Professor Atkinson very kindly sent the type collection for
examination. Under the ordinary high power of the microscope
the spores of both P. luddus and G. subperforatum appear to be
practically smooth. By the use of the oil-immersion lens vary-
ing degrees of apparent echinulation are to be made out in the
ordinary forms of P. lucidus while in the type collection of G.
subperforatum the spores do not have that appearance, although
Professor Atkinson states that by first boiling the spores in
potassium hydroxide solution the perforations in the spore walls
are faintly visible. I am convinced, however, that the echinu-
late appearance when present is not due to projections on the
outer wall, but, as Atkinson has said, to perforations in the
inner spore wall. An examination of the dozen or more col-
lections of P. lucidus in my own herbarium have given evidence
of a great variability in this character. Since G. subperforatum
is not otherwise to be distinguished from P. lucidus, it has seemed
best to consider the name as a synonym in this paper. Even
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 125
were the character constant one might well question the advisa-
bility of separating the species on a character that requires the
use of the oil-immersion lens for its detection.
This and the following species are included in the genus
Fomes by Saccardo, and many writers have followed his example.
Why this should be done is not clear, for both species are always
annual and the tubes are never stratified. The following
illustrations will aid in determination: Atkinson, Mushrooms
/. 188; Bot. Gaz. 46:/. 6., and Hard, Mushrooms/. 332.
61. P. Curtisii Berk. Hooker's Jour. Bot. Kew Gard. Misc.
i:101. 1849.
Pileus stipitate, reniform or flabelliform in outline, 3-12 x 3-20
X 0.7-2 cm., coriaceous-corky when fresh, corky when dry,
covered with a thin chestnut or reddish varnish that soon begins
1
to disappear, leaving the pileus yellowish or sometimes almost
white, glabrous, zonate or concentrically sulcate, the margin
rather thick, sometimes truncate; context in two layers, a yel-
lowish or pallid upper layer, rather soft in texture, and a brown-
ish lower layer next to the hymenium, firm or corky in texture,
the whole 0.5-1 cm. thick; tubes 0.3-1.2 cm. long, not at all
decurrent, the mouths white to brownish, mostly circular,
averaging 3-5 to a mm.; stipe always lateral, cylindrical, per-
sistently red-varnished and encrusted, sometimes bluish in
color, the context in two layers as in the pileus, 2-10 cm. long,
0.5-3 cm. thick; spores brown, ovoid to elliptic, smooth or
appearing minutely echinulate, with a heavy outer wall, 4.6-7.2
X 8.5-11.8 fx.
On and about stumps and trunks of trees. Rare.
This is typically a more southern plant and is rarely found
north of the Ohio River. - It is distinguished from the preceding
species by the yellowish color assumed by the mature pileus,
the change in color being due to the disappearance of the reddish
varnish. It is sometimes classed as a Fomes but is probably
never truly perennial. For illustration see Atkinson, Bot.
Gaz. 46:/. 1-3,
126 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
SPECIES DOUBTFUL OR EXCLUDED
The following species reported by either Morgan or Lea are
now believed to have been misdetermined, but the writer does
not know to what species the plants should be referred: P.
ovinus Schaeff. ex Fries; P. leucomelas Pers. ex Fries; P. lentus
Berk.; P. fragilis Fries; and P. badius Schw.
P. intybaceus Fries reported by Morgan is possibly a form of
P. giganteus, P. frondosus, or a closely related species.
P. phcEOxanthus Berk, was originally described from material
collected in Ohio by Sullivant. The type specimen is said to
be in fragments and the plant has never been collected since
Sullivant's time.
FOMES Fries, ex GiU,
Champ. Fr. 682. 1878. Fries; Nov. Symb. 31. 1851.
Plants typically perennial, epixylous, sessile (in our species) ;
pileus corky or more often woody in texture, often becoming
rimose, anoderm, or encrusted; context white, reddish, or brown-
ish, soft and punky to hard and woody; tubes as in Polyporus
except that they are arranged in definite or indefinite layers
corresponding to periods of growth of the plant, the mouths
circular or angular, never dsedaloid or irpiciform; spores white
or brown.
The genus Fomes includes all of the perennial forms which
have the tubes as in the genus Polyporus. Each season one
layer of tubes is produced and plants of the first season's growth
are likely to be referred to the genus Polyporus. The key to that
genus has been made to include a few such forms, the descrip-
tions of which are always to be sought in the genus Fomes. A
few species are so constantly annual iilduration that they might
perhaps better be included in the eenus Polyporus.
KEY TO THE SPECIES
Context white or only slightly colored {Species with wood-colcrred, flesh-colored,
or rose-colored context indvded here)
Context yellowish brown or darker
1
7
1. Sporophore small, scarcely more than 2 cm. broad; context white 2
1. Sporophore larger, more than 2 cm. broad; context whitish or somewhat
colored
3
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 127
2, Pileus entirely dark brown or black; plants growing only on the wood of
Abius and Hainameli^ 1. F. scutellatiis
2. Pileus not entirely black, the margin at least remaining white; plant
growing on the wood of other deciduous trees, often on structural
timber : S. F. ohiensis
3. Hymenium or context pinkish or reddish 4
3. Hymenium or context whitish or yellowish 5
4. Tubes more than 3 mm. long; plants usually growing on stumps and
trunks of Fraxinus 6. F. fraxineus
4. Tubes not more than 3 mm. long; plants usually growing on the wood of
coniferous trees 7. F. carjieus
5. Hyxaenium distinctly stratified, the strata of tubes separated by distinct
layers of context; mouths of the tubes angular, usually glistening , ,4- F. connatics
5. Hymenium indistinctly stratified or if somewhat distinctly so the layers not
separated by distinct layers of context; mouths of the tubes mostly circular,
not glistening 6
6. Plant growing on dead wood; usually of coniferous trees; mouths of the
tubes small, averaging 3-5 to a mm 5. F. jdnicola
6. Plant growing only on living Frax{nw5; mouths of the tubes rather large,
averaging 2 to a mm S. F. fraxinophilus
7. Pilei forming a densely imbricate, globose or cylindrical mass. . . .8. F. graveolens
7. Pilei not forming a densely imbricate, globose or cylindrical mass 8
8. Siu"face of the pileus not distinctly encrusted 9
8. Surface of the pileus distinctly encrusted 13
9. Context less than 5 mm. thick; sporophore often effuscd-reflexed or entirely
resupinate; growing usually on dead wood 9. F. conchattcs
9. Context more than 5 mm. thick; sporophore generally sessile; often growing
onliving trees 10
10. Sporophore found only on Robinia IL F. rimosus
10. Sporophore found on some other host 11
11. Tubes in the older layers distinctly white encrusted or stuffed '. . . . 12
11. Tubes in the older layers not distinctly white encrusted or stuffed 12. F, Everhartii
12. Surface of the pileus black, somewhat shining, and rimose; margin
rather thin and acute IS. F, igniarius
12, Surface of the pileus dull brown; margin thick and somewhat obtuse ....
14- F, nigricans
13. Encrusting layer thin, easily indented; plants annual or sometimes reviving
the second season but with the pileus distinct from and coming out below
that of the first season 17. F, lohatus
13. Encrusting layer thick and homy; plants strictly perennial 14
14. Plant growing only on species of Prunus 10, F, fulvus
14. Plant growing on some other host 15
15. Context hard and woody 13. F. igniarius
15. Context punky * . 16
16. Mouths of the tubes medium-sized, averaging 3 to a mm.; spores white
15. F. fomenlarius
16. Mouths of the tubes minute, averaging 5 to a mm; spores brown
16. F. applanatiis
9
128 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
1. F. scutellatus Schw. ex Cooke, Grevillea 14: 19. 1885.
Polyporus scutellatus Schw. Trans. Am. Phil. Soc. II. 4: 157.
1832.
Plants perennial, sessile, often attached by the apex of the
pileus; pileus dimidiate or circular, convex, 0.5-1.5 x 0.5-2 x
0.1-0.5 cm., corky when fresh, hard and woody when dry, dark
brown or black at least when mature, velvety, azonate or some-
what concentrically sulcate, margin rather thick, acute; context
white to wood-colored, corky, not more than 2 mm. thick; tubes
1-2 mm. long, indistinctly stratified, the mouths white to umber,
circular or subcircular, averaging 4-5 to a mm.
Chiefly on dead limbs of Alnus and Hamamelis. Rare.
This species is distinguished from F. ohiensis Berk, ex Murrill
by the habitat and the black surface of the entire pileus including
the margin. Specimens have been received from Mr. Claassen,
Cleveland, Ohio.
2. F. ohiensis Berk, ex Murrill, Bull. Torr. Bot. Club 30:
230. 1903.
Trametes ohiensis Berk. Grevillea i : 66. 1872.
Plants perennial, sessile, often attached by the vertex of the
pileus; pileus dimidiate or shield-shaped, convex to ungulate,
0.5-3 X 0.5-4 X 0.2-1 cm., soft-corky when fresh, hard and
woody when dry, at first pure white but becoming cinereous or
yellowish and often black at the base but the margin remaining
white, finely tomentose to glabrous, often zonate or concen-
trically sulcate, margin rather thick, acute or obtuse; context
white to wood-colored, soft-corky to woody, 1-3 mm. thick;
tubes 1-5 mm. long, often arranged in more or less definite rows,
indistinctly stratified in two to six layers, the mouths white,
circular, averaging 3-5 to a mm., the dissepiments almost as
thick as the diameter of the pores.
On dead wood of deciduous trees, especially on structural
timber. Common.
By its small size this species is separated from all perennial
forms except F. scutellatus Schw. ex Cooke. It differs from that
species in habitat and in the margin of the pileus always remain-
ing white.
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 129
3. F. fraxinophilus Peck ex Sacc. Syll. Fung. 6 : 172. 1888.
Polyporus fraxinophilus Peck, Ann. Kept. N. Y. State Mus.
35: 136. 1882.
Plants perennial, sessile or effused-reflexed, often imbricate;
pileus dimidiate, convex to compressed-ungulate, 2-25 x 3.5-40
X 1.5-10 cm., woody, white at first, becoming blackish and
often somewhat rimose with age, not encrusted, soon glabrous,
concentrically sulcate, margin thick, obtuse or acute; context
white to cinnamon wood-color, corky or woody, 0.5-1 cm. or
more thick; tubes 2-3 mm. long, indistinctly stratified in many
layers, the mouths white to cinereous or yellowish, circular,
averaging 2-3 to a mm., the walls thick and entire; spores white,
smooth, ellipsoid to ovoid or pyriform, 5-6,3 x 7.3-8 //.
Growing only on living trunks of Fraxinus. Common.
In habitat the species corresponds closely to F. jraxineus
Bull, ex Cooke, from which it differs in the entire absence of any
rosy or reddish colors and in being always perennial. An
excellent illustration is given by Hard (Mushrooms/. 350).
4. F. connatus Weinm. ex Gill. Champ. Fr. i: 684. 1878.
Polyporus connatus Weinm. Fl. Ross. 332. 1836. P. con-
natus Fries, Epicr. Syst. Myc. 472. 1838.
Plants perennial, sessile or effused-reflexed, sometimes imbri-
cate; pileus dimidiate, convex, 2-10 x 3-15 x 0.5-4 cm., corky
when fresh, somewhat woody when dry, whitish, cinereous,
or slightly yellowish, sometimes blackish toward the base, not
encrusted, velvety-tomentose to glabrous, usually azonate,
margin thick, acute or obtuse; context white or pallid, punky
to soft corky, 0.3-1 cm. thick; tubes 1.5-5 mm. long, distinctly
stratified, the different strata separated from each other by
a thin layer of context, the mouths whitish to yellowish, glis-
f
tening, angular, averaging 4-5 to a mm., the walls entire to
sHghtly dentate; spores (teste Bresadola) white, globose, 3-4 m
in diameter.
Growing on living deciduous trees, more often at the bases
of species of Acer, and frequently covered with moss. Common.
The distinguishing characters are the habitat, the layers
of context interposed between successive layers of tubes, and
the glistening mouths of the tubes. In but one other species
of Fames do we find the second character developed and that
[Vol. 1
130 ANNALS OF THE MISSOURI BOTANICAL GARDEN
is in F. applanatus Pers. ex Wallr. That species always grows
on old logs and stumps and has a rusty brown context.
4
Bresadola and Murrill regard F. populinus Schum. ex Cooke,
to be the same plant as this species. This may be the case
but the figure of F. connatus in Fries' Icones' (/. 185) represents
our plant much better than the figure of F. populinus in 'Flora
Danica' (pi. 1791). Most of the specimens distributed in
exsiccati in both this country and Europe are under the former
name and that one is here given preference. A study of the
types of both species should show whether they are the same
or not, but from the evidence at hand our plants must be
referred to F. connatus.
5. F. pinicola Sw. ex Cooke, Grevillea 14: 17. 1885.
Boletus pinicola Sw. Sv. Vet. Akad. Handl. 88. 1810. Pol-
porus pinicola Fries, Syst. Myc. i:372. 1821.
Plants perennial, sessile; pileus plane to convex, rarely ungu-
late, dimidiate, 4-15 x 6-20 x 3-10 cm., woody and rigid, gray-
ish to black, partly or entirely covered with a reddish gluten
that forms a crust over the surface, glabrous, sometimes con-
centrically sulcate, margin thin or thick, often obtuse; context
pallid or wood-colored, corky to woody, 0.5-2 cm. thick; tubes
3-5 mm. long, distinctly or indistinctly stratified, the mouths
white to umber, circular, averaging 3-5 to a mm., the walls
thick and entire.
On dead wood, usually of coniferous trees.
Distinguished from closely related species by the resinous,
somewhat sticky, reddish crust found on the pileus. The
plant is common wherever coniferous woods are found. Hard
(Mushrooms /. 348) gives a photograph of it but does not state
that he ever collected it in Ohio.
6. F. fraxineus Bull, ex Cooke, Grevillea 14: 21. 1885.
Boletus fraxineus Bull. Herb. Fr. pi. 433. f. %. 1789. Poly-
poTus fraxineus Bull, ex Fries, Syst. Myc. i:374. 1821.
Plants annual or perennial, sessile, sometimes imbricate;
pileus dimidiate, 4-10 x 6-15 x 0.6-2 cm., corky when fresh,
rigid and woody when dry, light colored, always with reddish
or reddish brown stains, or altogether reddish, encrusted with
a thin hard crust, minutely velvety to glabrous, more or less
zonate, margin thin or thick, acute; context floccose-punky
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 131
to corky, whitish or palhd when dry, tinged pinkish or flesh-
colored when fresh, 0.2-2 cm. thick; tubes 2-6 mm. long,
usually in a single layer but sometimes stratified, mouths
whitish, palHd, or flesh-colored, circular or subcircular, averaging
4-6 to a mm., the dissepiments rather thick and entire; spores
(teste Murrill) subglobose, smooth, subhyaline, 5-6 x 6-7 /i.
Usually found on living Fraxinus but sometimes on other
hosts. Rare.
The habitat, the reddish blotches on the pileus, and the pink-
ish hymenium and context in fresh specimens will identify
the plant. But three collections are known from Ohio; one
by Morgan, one by W. G. Stover near Columbus, in 1910, and
one near Camden, Ohio, by the writer, in 1912. All of these
collections are of the annual form.
7. F. cameus Nees ex Cooke, Grevillea 14: 21. 1885.
Polyporus cameus Nees, Nova Acta Acad. Leop. Carol. 13:
pi 3. 1827.
Plants annual or perennial, sessile; pileus dimidiate, 1.5-5 x
1.5-10 X 0.3-1.5 cm., soft-corky when fresh, firmer when dry,
pinkish or rosy, sometimes blackish with age, velvety to gla-
brous, usually azonate, margin thin and acute; context pinkish
or rosy, floccose or punky to soft-corky, 0.2-1 cm. thick; tubes
0.5-3 mm. long, usually in a single layer but sometimes strati-
fied, mouths pinkish or rosy, circular or subcircular, averaging
3-5 to a mm., dissepiments thick and entire.
Usually on wood of coniferous trees. Rare.
The species will be recognized by the uniform pinkish color
of the whole plant. The color is well retained on drying.
Specimens are in the herbarium of the New York Botanical
Garden, collected by James, in Ohio. Authorities disagree as
to the identity of F. cameus and F. roseus Fries ex Cooke.
8. F. graveolens Schw. ex Cooke, Grevillea 13:118. 1884.
Boletus graveolens Schw. Syn. Fung. Car. 97. 1822. Poly-
porus conglohatus Berk. Hooker's Lond. Jour. Bot. 4:303. 1845.
Plant composed of numerous overlapping pilei arising from
a central solid core and forming a more or less globose or cylin-
drical mass 5-12 cm. in diameter; pilei not more than 2 cm.
broad, but connate laterally, corky when fresh, rigid and firm
when dry, grayish browm to dull cinnamon-brown, becoming
[Vol. 1
132 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
black in weathered specimens, slightly encrusted; pulverulent
to glabrous, azonate or marked with fine grayish zones, margin
thick, deflexed and almost concealing the pores; context fulvous
to golden brown rust-colored, floccose-fibrous, 1-4 mm. tliick;
tubes 2-4 mm. long, the mouths grayish, hoary brown, or
umber, circular, averaging 3-4 to a mm., the dissepiments
thick and entire.
On logs or trunks of deciduous trees, especially Fagus, Quercus,
and Acer. Rare.
A characteristic plant that will be recognized at sight. It
is commonly known as "sweet knot" from the sweet, powerful
odor that it is said to give off. The writer has made four
different collections of this rare plant in different stages of
growth but has never been able to detect the sHghtest semblance
of a sweet odor. The plant is exceptionally well illustrated by
Lloyd (Myc. Notes, Polyp. Issue 3: /. 367-68; Syn. Stip.
Polyp. /. 455), and Hard (Mushrooms /. 334). The first and
the last of the figures cited are upside down.
^
9. F. conchatus Pers. ex Gill. Champ. Fr. i:685. 1878.
J
Boletus conchatus Pers. Obs. Myc. i:24. 1796. Polyporus
conchatus Fries, Syst. Myc. i:376. 1821.
Plants perennial, sessile, or more often effused-reflexed and
frequently entirely resupinate; pileus dimidiate to conchate,
0-7 x 4-12 X 0.2-3.5 cm., woody, grayish brown, yellowish
brown or black, rarely encrusted, tomentose at least on the
margin, becoming glabrous behind, zonate or concentrically
sulcate and sometimes somewhat rimose, margin thin, mostly
acute; context yellowish brown to dark brown, woody, 1.5-3
mm. thick; tubes 1-2 mm. long, indistinctly stratified, mouths
fulvous to dark brown, usually glistening, circular or subcircular,
averaging 4-6 to a mm.
On dead wood, rarely on living trees. Common.
The plant is most frequently found entirely resupinate on
old oak logs. Distinctly sessile forms are sometimes found,
especially on living trees. The hymenium usually has a silky
luster when viewed in changing positions, and on the whole
the plant is so characteristic that when once recognized, the
collector usually has no trouble with subsequent collections
notwithstanding the fact that the species often presents great
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 133
differences in size and habit. In Europe the plant is usually
known as F. salicinus Pers. ex Gill, and it was so reported from
Ohio by Morgan. It is entirely different from all other species
of Fomes except F. fulvus Scop, ex Gill, and F. rihis Schum.
ex Fries in the thin pileus, often conchate in form and with a
concave hymenium. Usually the pileus is not more than 1
cm. thick. F. fulvus Scop, ex Gill, is distinct in its habitat
as is also F. rihis Schum. ex Fries.
10. F. fulvus Scop, ex Gill. Champ. Fr. i: 687. 1878.
Boletus fulms Scop. Fl. Carn. 2:469. 1772. [2nd ed.] Poly-
porus fulvus Fries, Epicr. Syst. Myc. 466. 1838.
Plants perennial, sessile, effused-reflexed or entirely resupinate;
pileus dimidiate, convex, 0-4 x 3-8 x 0.5-3 cm., woody, fulvous to
ferruginous when young, becoming grayish black or black
in age, encrusted, minutely velvety to glabrous, sometimes
sulcate, margin rather thick, acute or obtuse; context dark
brown, woody, 3-8 mm, thick; tubes 2-4 mm. long, rather
distinctly stratified, the mouths circular to slightly angular,
grayish brown to tawny, averaging 4-5 to a mm., dissepiments
rather thick, entire.
Growing only on wood of species of Prunus. Not common.
One should have no trouble in identifying this species if
the habitat is taken into consideration as it is the only perennial
form that grows on Prunus. Morgan reported it under the
name of F. supinus Schw.
11. F. rimosus Berk, ex Cooke, Gre^dllea 14: 18. 1885.
I .
Polyporus rimosus Berk. Hooker's Lond. Jour, Bot. 4:54.
1845. Pyropolyporus rohiniae Murrill, Bull. Torr. Bot. Club
30:114. 1903.
Plants perennial, sessile; pileus dimidiate, convex to ungulate,
3-20 x 6-30 X 1.5-10 cm., woody, at first fulvous, becoming
dark brown or black, not encrusted, velvety in young speci-
mens, glabrous and very rimose in old plants, concentrically
sulcate, margin thick or thin, obtuse or acute; context fulvous
to rusty brown, woody, 0.5-3 cm. thick, zonate; tubes 1-5 mm.
long, indistinctly stratified in many layers, the mouths fulvous
to rusty brown, circular, averaging 5-6 to a mm., walls rather
thick and entire; spores brown, smooth, globose, 4-5m in
diameter.
.r
134 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
Growing only on living trunks of Robinia. Common.
The type locality for F. rimosus is given by Berkeley as the
Swan River, Australia, and not Demerara and the Cape of
Good Hope, as cited by Saccardo and by Murrill. If the speci-
mens Murrill examined are from the two latter places, it is still
possible that our plants belong under F. rimosus. Our species
also occurs in South Africa as specimens examined from that
locality agree well with our plants.
The plant is never found on any other host than the locust
tree. This will distinguish it from all of its allies. Its closest
relatives appear to be F. Everhartii EUis & Gall, and F. igniarius
L. ex Gill. The plant is well illustrated by Hard (Mushrooms
/. 5^7), and by von Schrenk (Ann. Rept. Mo. Bot. Gard. 12: pi 2),
12. F. Everhartii Ellis & Gall.^
Mucronoporus Everhartii Elhs & Gall. Jour. Myc. 5:141.
1889.
Plants perennial, sessile or decurrent; pileus dimidiate, convex,
rarely ungulate, 2.5-10 x 4-20 x 2-6 cm., woody, entirely
fulvous when young but becoming grayish brown or black
and rough and rimose with age, velvety when young, glabrous
when mature, scarcely encrusted, concentrically sulcate with
age, margin thin or thick, acute or obtuse, usually remaining
fulvous in color; context fulvous to rusty brown, shining (at
least in herbarium specimens), zonate, woody, 1-4 cm. thick;
tubes 3-6 mm. long, indistinctly stratified, tubes of the older
layers sometimes partly stuffed with mvcelium. the mouths con-
with the
5 to a mm.,
the walls rather thin but entire, sometimes ghstening; spores
distinctly brown, smooth, globose, 4-5.3 m in diameter.
On living trees, usually of Quercus. Not uncommon.
Distinguished from F. igniarius L. ex Gill, and F. nigricans
Fries ex Gill, by the absence of the distinct encrustation or
V
stuffing of the tubes in the old layers, by the more shining
context; the somewhat thinner dissepiments, the hyaline spores,
and the absence of a distinct crust on the pileus. The two
J F. Everhartii was originally described under the genus Mucronoporus and as
far as I have been able to find, no specific statement of transfer to the genus Fomes
was ever made. At the present time I have not been able to satisfy myself as to
who was the first to make (unknowingly, it seems) the new combination, and there-
fore I do not know to whom credit for the transfer should be given.
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 135
species are closely related, however, and without the spores
it is sometimes difficult to decide between them. The species
differs from F. fomentarius L. ex Gill, and F. applanatus Pers.
ex Wallr. in the unencrusted pileus, the woody context, and the
short tubes.
13. F. igniarius L. ex Gill. Champ. Fr. i:687. 1878.
Boletus igniarius L. Sp. Plant. 1176. 1753. Polyporus igni-
arius Fries, Syst. Myc. i:375. 1821.
Plants perennial, sessile; pileus dimidiate, convex to some-
what ungulate, 2.5-11 x 4-25 x 1.5-12 cm., woody, grayish
black, or entirely black, encrusted, sometimes somewhat rimose
in age, glabrous, concentrically sulcate in older plants, margin
rather thin, acute, usually grayish in growing specimens;
context rusty red or rusty brown, scarcely shining, zonate,
woody, 0.5-4 cm. thick; tubes 2-5 mm. long, usually indis-
tinctly stratified, the older layers becoming distinctly whitish
encrusted, the mouths circular, grayish brown to dark brown,
averaging 4-5 to a mm., the walls thick and entire; spores
(teste Romell) hyaline, subglobose, 5-7.5x4-7 m, often 1-guttate.
On trunks of living deciduous trees. Not common.
In no other species is the stuffing or encrusting of the tubes
by a whitish substance so evident as in this and the next one.
In F. Everhartii Ellis & Gall, the tubes appear to be sometimes
filled with a whitish mycelium but the character is scarcely
evident except on close examination, while in F. igniarius
and F. nigricans Fries ex Gill, in sections through the hymenium
the whitish encrustation is plainly visible, and seems to be a
I
distinguishing character. The plant is further to be distin-
guished from F. Everhartii by the hyaline spores, and the
thicker dissepiments. The pores are somewhat smaller, but
in measuring them the thick dissepiments are included, so
that the number per mm. is about the same in the two species.
From F. fomentarius L. ex Gill, and F. applanatus Pers. ex
Wallr. the species is separated by the more woody context,
the thinner crust, and the much shorter tubes, as well as by
the hyaline spores. In F. igniarius the pileus is darker in color
and is usually much more rimose than in F. nigricans. For
I
illustrations see Atkinson, Cornell Univ. Agr. Exp. Sta, BuL
193:/. 7S-4'
136 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
14. F. nigricans Fries ex GilL Champ. Fr., Hymen, i: 685,
1878.
Polyporus nigricans Fries, Syst. Myc. i:375. 1821.
Plants perennial, sessile; pileus dimidiate, convex to ungu-
late, distinctly triangular in cross-section, 5-10 x 7-13 x 2-7
cm., woody, dull brown or becoming brownish black, not en-
crusted, smooth or cracking somewhat in age but scarcely
rimose, azonate or with one or two concentric furrows, the
margin thick, acute or obtuse, with a broad ferruginous band;
context rusty brown, zonate, woody, 0.6-2 cm. thick; tubes
2-7 mm. long, distinctly or indistinctly stratified, becoming dis-
tinctly white encrusted or stuffed in the older layers, the mouths
dark brown, circular, minute, averaging about 5 to a mm.,
the walls thick and entire; spores white, subglobose or globose,
6.5^ in diameter.
On trunks of trees, especially on Betula. Not common.
I have one collection of this fungus from W. A. Kellerman.
The species has been confused with the preceding one from
which it differs in the smoother and differently colored pileus
and in being more decidedly triangular in cross-section. The
best illustration is that given by Boudier (Ic. Myc. i: pi 155).
15 F. fomentarius L. ex Gill. Champ. Fr. 1:686. 1878.
Boletus fomentarius L. Sp. Plant. 1176. 1753. Polyporus
fomentarius Fries, Syst. Myc. i:374. 1821.
Plants perennial, sessile; pileus dimidiate, convex to strongly
ungulate, 3.5-15 x 6-20 x 2-9 cm., hard and woody, grayish to
cinereous, brownish, or black, covered with a thick horny crust
that appears black and shining when cut, glabrous, smooth,
never rimose, zonate or concentrically sulcate, margin thick
and obtuse; context fulvous to ferruginous, never shining,
punky to soft-corky, zonate, 0.3-3 cm. thick; tubes 0.5-2.5
cm. long, rather distinctly stratified, mouths grayish to cin-
namon, averaging 3 to a mm., the walls thick and entire.
On living deciduous trees. Not common.
Distinguished from all of the preceding species by the punky
or soft-corky context and the usually longer tubes. Most
closely related to F. applanatus Pers. ex Wallr. but distinguished
from it by the much longer pores and the hyaline spores. For
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 137
illustrations see Kellerman, Journ. Myc. 9: pi 3., and White,
Hymen. Conn. pi. 35. f. 2.
16. F. applanatus Pers. ex Wallr. Crypt. Fl. Ger. 2:591.
1833.
Boletus applanatus Pers. Obs. Myc. 2: 2. 1799. Polyporus
applanatus Fries, Epicr. Syst. Myc. 465. 1838. P. leucophcBus
Mont. Syll. Crypt. 157. 1856.
Plants perennial, sessile; pileus dimidiate, convex or plane,
not ungulate, 3-30 x 5-50 x 1.5-7 cm., woody, usually grayish
becoming brownish or blackish, glabrous, covered with a thick
horny crust, zonate or concentrically sulcate, margin thin or
thick, acute or obtuse; context dark ferruginous brown, floccose
to soft corky, 0.6-2 cm. or more thick; tubes 0.5-1.5 cm. long,
distinctly stratified after the first season, the strata separated
by thin layers of context, mouths whitish to umber, darker
when bruised, cu-cular, minute, averaging 5-6 to a mm.
On dead wood of deciduous
Common
This is our most common species of Fomes and may be found
in every woodlot, usually on stumps or old logs. It is dis-
tinguished from F. fomentarius L. ex Gill, by the more applanate
pileus and the minute mouths of the tubes. For
see Atkinson, Mushrooms /. 15., 'White, Hymen. Conn, pi
f. L, and Atkinson, Cornell Univ. Agr. Exp. Sta. Bui
17. F. lobatus Schw. ex Cooke, Grevillea 14:18. 1885.
Polyporus lohatus Schw. Trans. Am. Phil. Soc. 11.4:157. 1832.
P. reniformis Morgan, Journ. Cin. Soc. Nat. Hist. 8:105. 1885.
Plants annual, frequently reviving for two or three years
but the second year's growth distinct from and coming out
below that of the first year, sessile or more often appearing
substipitate; pileus dimidiate or reniform, plane, depressed, or
somewhat convex, never ungulate, 4-12 x 4-15 x 1-3 cm.,
corky or somewhat flexible when growing, usually umber to
yellowish or dark rusty brown, glabrous, covered with a thin,
easily indented crust, zonate or concentrically sulcate, margin
thin and acute; context dark rusty brown, soft and floccose to
punky, 0.3-1 cm. thick; tubes 0.4-1 cm. long, not stratified,
mouths circular or subcircular, white, yellowish or umber-
ici» ANNALS OF THE MISSOURI BOTANICAL GARDEN
brown, darker when bruised, averaging 4 to a mm., walls rather
thin but entire.
On dead wood of deciduous trees. Common.
This species is easily separated from F. applanatus Pers. ex
Wallr. in that it is not perennial, and in that, if the plant revives
the second year, the pileus comes out below that of the first
year, and the latter persists as a dead decaying pileus. The
second difference is in the character of the encrusting layer
of the pileus. In F. applanatus Pers. ex Wallr. the crust is hard
and horny and one cannot indent it with the thumb nail, while
in F. lohatus the crust is thin, and often becomes cracked and
brittle when old, but is always rather soft and easily indented.
TRAMETES Frie.-, Gen. Hymen. 11. 1836.
Plants annual or perennial, epixylous, sessile; pileus corky
or woody in texture, small or medium sized; context white or
brown (never red), descending into and forming the walls of
the tubes; tubes typically appearing sunken into the context
to unequal depths so that their bases are not in a continuous
straight line; mouths circular or angular, never breaking up
mto teeth and rarely showing a dacdaloid tendency.
One species here included in the genus is perennial, all
others are annual. The chief generic distinctions are the uneq...
depths to which the tubes are immersed in the context, and
the homogeneous texture of the context and trama. The first
distinction is often not apparent except on very close examina-
tion, and at times appears to break down entirely. Conse-
quently, students will meet with some diflSculty at times in
deciding between the two genera, Trameies and Polyporus.
the
KEY TO THE SPECIES
Context white or whitish I
Context brown or brownish a
1. Pileus densely hirsute or hispid 7. T Peckii
1. Pileus slightly pubescent to glabrous ................'. .'..... 2
2. Mouths of the tubes minute, averaging 4-6 to a mm .' .T. robiniophila^
2. Mouths of the tubes larger, averaging 1-3 to a mm 3
8. Pileus rather large; context more than 5 mm. thick; plant growing only
on Salix • ™, ,
„ _,., S. T. siiaveolens
6. l^ileus small, sometimes mostly resupinate; context less than 5 mm.
thick; foxmd on some other host 4
J For description of this plant see p. 104 under the genus Polyporus.
1914]
OVERHOLTS — THE POLYPORACEiE OF OHIO 139
4. Hymenium light brown in color 4. T. malicola
4. Hymenium white or whitish
5. Pileus white or light colored; mouths of the tubes averaging 1-2 to a mm.
5
1. T. sepium
5. Pileus cinnamon-brown; mouths of the tubes averaging about 3 to a mm.
f . T. serialis
6. Sporophore woody, perennial; hymenium bright yellowish brown in color;
mouths of the tubes often somewhat daedaloid; growing only on
Pinus 8. T, Pint
6, Sporophore coriaceous or corky; hymenium white or dull brown; growing
on wood of deciduous trees 7
7. Pileus hirsute or hispid 8
7. Pileus finely tomentose or glabrous * 9
8. Mouths of the tubes large, averaging 1 to a mm.; pileus more than 4 mm.
thick 7. r. Peckii
8. Mouths of the tubes medium sized, averaging 2-3 to a mm.; pileus less
than 4 mm. thick 6, T. rigida
9. Context less than 1 mm. thick 5. T. mollis
9. Context more than 1 mm. thick ^. T, malicola
1. T, sepium BerL Hooker^s Lond. Jour. Bot, 6:322. 1847.
Plants annual, sessile or semirespuinate, imbricate or single;
pileus dimidiate, 0.7-1 x 0.8-2.5 x 0.2-0.7 cm., flexible when
fresh, corky when dry, grayish to pallid or wood-colored, mi-
nutely tomentose to glabrous, azonate, margin thin and acute;
context white or pallid, tough when fresh, soft-corky when
dry, less than 1 mm. thick; tubes 2-5 mm. long, mouths w^hite
or pallid; circular or rarely angular or sinuous^ large, averaging
almost 1 to a mm., the dissepiments thick and always entire;
spores (teste Murrill) oblong, smooth, hyaline, 12 x 5 //.
On fence posts, pickets, and other structural timber or dead
wood.
Distinguished from T, serialis Fries by the short tubes, the
whitish color of the pileus, and by the much larger mouths of
the tubes; from T. rigida Berk. & Mont, by the lighter colored
context, and the larger tube mouths.
2. T. serialis Fries, Hymen. Eur. 585. 1874. [2nd ed.]
Polyporus serialis Fries, Syst. Myc. i:370. 1821.
Plants annual, sessile, effused-reflexed, or resupinate; pileus
dimidiate, 0-1 x 1-4 x 0.3-0.8 cm., corky when fresh, hard
and firm when dry, cinnamon-brown to coffee brown, glabrous,
zonate, margin rather thick but acute; context white, fibrous,
not more than 1 mm. thick; tubes 2-6 mm. long, the mouths
[YOL. 1
140 ANNALS OF THE MISSOURI BOTANICAL GARDEN
white or slightly discolored, sometimes slightly glistening,
circular to angular, averaging 3 to a mm., the walls firm and
entire; spores (teste Bresadola) hyaline, elongate, 7-10 x 3-3.5 fx.
On dead wood. Rare.
The white pores and the internally white tubes contrast
strongly with the rich brown color of the pileus. It is distinct
from T. rigida Berk. & Mont, in the glabrous, thicker pileus.
From T. sepium Berk, it differs in the much smaller pores
and the brown pileus; T. malicola has no white color in the tubes-
and the dissepiments are much thicker.
3. T. suaveolens L. ex Fries, Syst. ]\Iyc. i:366. 1821
Boletus suaveolens L. Sp. Plant. 1177. 1753.
Plants annual, sessile; pileus dimidiate, 3-9 x 6-14 x 1-3
cm., corky when fresh, firm and rigid when dry, white to grayish
or sHghtly yellowish, finely villous-tomentose to glabrous,
azonate, margin thin or thick, acute; context white or palhd,
compact-corky to somewhat indurate, 0.5-2 cm. thick; tubes
0.2-1.5 cm. long, the mouths white or cinereous, circular to
:htly
3 to a mm
On dead or diseased Salix, Rare.
Distinguished from T. Peckii Kalchbr. by the prevailing
whitish color and the more nearly glabrous pileus.
4. T. malicola Berk. & Curt. Journ. Acad. Nat. Sci. Phil.
11.3:209. 1856.
Plants annual or reviving for two or three seasons, effused-
reflexed
0-1 x 1-5
X 0.3-0.8 cm., coriaceous and leathery when fresh, corky when
dry, avellaneous to cinnamon-brown or wood-colored, azonate,
margin thick but acute; context wood-brown or lighter, soft-
corky, 2-5 mm. thick; with a distinct pleasant odor when fresh;
tubes 2-5 mm. long, sometimes indistinctly stratified in two or
three layers, mouths wood-colored to cinnamon-brown, circular
to angular or somewhat sinuous, averaging about 2 to a mm.,
the walls thick and entire; spores white, smooth, oblong, 2.8
-3.5x7.5-10//.
Growing on dead wood of deciduous trees, especially species
of Acer. Common.
Entirely distinct from T. sepium Berk, in the semiresupinate
1914]
OVEKHOLTS — THE POLYPORACE^ OF OHIO 141
habit of growth, the prevaiUng dull brown color of both hyme-
nium and pileus, and the smaller-mouthed tubes. In this last
respect the plant more nearly approaches T. serialis Fries and
T. rigida Berk. & Mont. From the former it is separated by
the browner color of the hymenium, the lighter color of the
pileus, the internally brown tubes, and the sHghtly larger
and more irregular mouths. From the latter it differs chiefly
in the more glabrous and less developed pileus and the longer
tubes.
The type specimens of T. vialicola were collected on the trunk
of an apple tree by Schweinitz and referred by him to P. popu-
linus Fries. Murrill has placed the name as a doubtful syno-
nym for P. galactinus Berk. The writer has not examined the
type of T. malicola, but our plants bear no resemblance to
either P. populinus Fries or P. galactinus Berk. Our plants
w^ere determined by Lloyd and by Bresadola.
5. T. mollis Sommerf. ex Fries, Hymen. Eur. 585. 1874.
Daedalea mollis Sommerf. Suppl. Fl. Lapp. 271. 1826.
Plants annual or rarely reviving, rarely sessile, more often
eifused-reflexed or entirely resupinate; pileus dimidiate or
elongate, 0-2.5 x 1-4 x 0.1-0.5 cm., coriaceous to rigid, umber-
brown to almost black, finely tomentose to glabrous, zonate or
multizonate, margin thin and acute; context Hght brown,
fibrous, less than 1 nmi. thick; tubes 2-3 mm. long, rarely in
two or three layers, mouths light brown or grayish, subcircular
to somewhat angular, often becoming sinuous or labyrinthi-
f orm, averaging 1-3 to a mm. ; spores (teste Bresadola) elongate-
ellipsoid, smooth, hyaline, 9-11 x 4-4.5 ju.
On dead wood. Rare.
The species differs from T. rigida Berk. & Mont, in the
distinctly brown and almost glabrous pileus. From T. serialis
Fries it differs in the light brown context, the much thinner
pileus and the usually larger and more irregular pores. The
context is much thinner than in T. malicola Berk. & Curt, and
the general color is decidedly different.
6. T. rigida Berk. & Mont. Ann. Sci. Nat. III. ii:240.
1849.
Plants annual or rarely reviving, sessile, effused-reflexed or
entirely resupinate, sometimes imbricate; pileus dimidiate,
142 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
0-3 X 2-6 X 0.1-0.3 cm., coriaceous when fresh, coriaceous or
rigid when dry, cinereous to yellowish or slightly brownish,
hirsute to hispid, usually zonate, sometimes with multicolored
zones, margin very thin and acute; context Hght umber, fibrous,
0.5-3 mm. thick; tubes not more than 1 mm. long, the mouths
white or brownish, circular to somewhat angular, averaging 2-3
to a mm., the walls rather thin but entire.
On dead wood. Not common.
Distinguished from all of its allies in the hirsute or hispid
pubescence of the pileus. The pileus is thin and coriaceous
and more nearly resembles the thin coriaceous species in Poly-
porus.
7. T. Peckii Kalchbr. Bot. Gaz. 6: 274. 1881.
Plants annual, sessile or effused-reflexed ; pileus dimidiate,
1.5-6 x 2.5-12 X 0.5-2 cm., somewhat coriaceous when fresh,
firm and rigid when dry, yellowish brown or reddish brown,
densely hirsute or hispid, concentrically sulcate at times, margin
thick or thin, acute; context light brown, fibrous, soft and spongy
to firm and woody, 1-10 mm. thick; tubes 2-10 mm. long, the
mouths dull brown or grayish brown, angular to irregular,
averaging about 1 to a mm.; spores (teste Murrill) oblong or
sHghtly curved, smooth, hyaline, 11-13 x 3.5-4 ju.
On dead wood of Populus, Liriodendron, and Salix. Sep-
tember to December. Frequent.
Easily recognized by the densely hirsute or hispid pubescence,
the large pores, and the habitat. In Europe the species is known
as T. hispida Fries.
8. T. Pini Thore ex Fries, Epicr. Syst. Myc. 489. 1838.
Boletus Pini Thore, Essai Chlor. Dep. Land. 487. 1803.
Plants perennial, sessile or effused-reflexed; pileus dimidiate,
often ungulate, 3-15 x 5-20 x 1-6 cm., woody, yellowish brown
to reddish brown or becoming black, the growing margin hir-
sute to tomentose, glabrous behind, zonate or concentrically
sulcate, margin usually thick and somewhat obtuse; context
yellowish brown to rusty brown, corky to woody, not more than
5 mm. thick; tubes 2-7 mm. long, indistinctly stratified, the
mouths usually golden brown, subcircular to daedaloid and laby-
rinthiform; spores (teste Bresadola) hyaUne, subglobose,
5-6 x 4-5 a.
1914]
OVERHOLTS — THE POLYPORACE.E OF OHIO 143
On coniferous wood. Rare.
The bright color of the hymenium usually contrasts strongly
dth the darker colors of the upper surface. P. piceinus Peck
= Trametes Abietis Karst.), which by some is regarded as a
form of T. Pini, has never, to the writer's knowledge, been
collected within the state.
SPECIES DOUBTFUL OR EXCLUDED
r. nivosus Berk, was erroneously reported from Ohio by
Morgan. It is a tropical and subtropical species.
DAEDALEA Pers. ex. Fries,
Syst. Myc. i: 331. 1821; Pers. Syn. Fung. 499. 1801.
Plants annual or rarely reviving for two or three years, ses-
sile or effused-reflexed, growing on wood; pileus coriaceous to
corky in texture, not encrusted; context white or whitish,
fibrous or corky; hymenium typically daedaloid or labyrinthi-
form, but sometimes poroid, irpiciform or lamellate; spores
white.
KEY TO THE SPECIES
Pileus small, thin and coriaceous, hirsute or villous; hymenium at first sin-
uous and dsedaloid but soon breaking up into teeth 1. D. unicolor
Pileus rather large and thick, corky, minutely velvety or glabrous; hyme-
nium poroid, dsedaloid, or somewhat lamellate but never breaking up
into teeth 2
2. Mouths of the tubes less than 1 mm. broad 2. D, amhigua
2. Mouths of the tubes more than 1 mm. broad 3
3. Pileusless than 1.5 cm. thick; walls of the tubes thin; plant found abun-
dantly on Salix 5. D. confragosa
3. Pileus more than 1.5 cm. thick; walls of the tubes thick; plant growing
on Quercus and Castanea ^. D
qv£rana
I. D. unicolor Bull, ex Fries, Syst. Myc. i: 336. 1821.
Boletus unicolor Bull. Herb. Fr. pi. 408. 1788.
Plants annual or sometimes the marginal hyphse reviving and
continuing growth the second year, sessile, or effused-reflexed,
imbricate; pileus dimidiate to flabeUiform, 0.5-5 x 2-8 x 0.2-
0.5 cm., coriaceous, white to cinereous or light
times green from a covering of algse, villous or hirsute, zonate or
concentrically furrowed, margin thin, acute, sterile below; con-
text white or pallid, fibrous, less than 1 mm. thick; tubes 1-4
mm. long, the mouths white to cinereous or umber, at first
10
brown, some
[Vol. 1
144 ANNALS OF THE MISSOURI BOTANICAL GARDEN
dsedaloid and sinuous, but soon breaking up into teeth — though
retaining the sinuous character at the margin of the pileus
averaging about 2 to a mm.
On dead wood. Common.
This plant may at first prove puzzUng to the collector, as it
was to me when first collected, for the thin, flexible pileus and
the usually toothed hymenium indicate a close relationship with
the thin coriaceous species of Polyporus, or even with Irpex.
But the pores are decidedly sinuous, at least in young plants.
The thin pileus and the hirsute or villous pubescence separate
the species from other members of the genus.
2. D. ambigua Berk. Lond. Jour. Bot. 4: 305. 1845.
Trametes laciea Berk. Hooker's Lond. Jour. Bot. 4 : 305. 1845.
Plants annual or rarely reviving for two or three years, sessile,
sometimes appearing substipitate; pileus dimidiate to reniform,
3-14 X 5-20 X 0.3-1.5 cm., sUghtly flexible when fresh, corky
when dry, pure white to umbrinous, sometimes purpUsh black
at the base, minutely velvety to glabrous, azonate or subzonate
on the margin, margin rather thin, acute; context white or pal-
lid, floccose-punky to corky, 0.2-1 cm. thick; tubes 2-4 mm.
long, sometimes stratified in two or three layers, mouths whit-
ish or yellowish, circular to sinuous and dsedaloid, never lamellate,
in transverse direction, walls rather
thick and entire.
On stumps and trunks of deciduous trees. Common.
Distinguished from D. confragosa Bolt, ex Fries by the white
color, the white context, the smaller pores and the habitat.
Hard (Mushrooms /. 355-56) gives excellent illustrations of the
plant.
3. D. confragosa Bolt, ex Fries, Syst. Myc. i : 336. 1821.
Boletus confragosus Bolt. B.mt. Fung. Suppl. 3:160. 1791.
Lenzites Cratcegi Berk. Hooker's Lond. Jour. Bot. 6 : 323. 1847.
Plants annual, sessile; pileus dimidiate, 2-10 x 3-15 x 0.2-1.5
cm., slightly flexible to rigid, grayish or cinereous, rarely slightly
brownish, minutely tomentose to glabrous, zonate, margin thin
and acute; context whitish, floccose to corky, 0.2-1 cm. thick;
tubes 0.1-1 cm. long, mouths whitish to cinereous, sometimes
shghtly reddish, darker when bruised, subcircular at times but
usually sinuous, dsedaloid, or labyrinthiform, sometimes becom-
mm
1914]
OVERHOLTS — THE POLYPORACE.E OF OHIO 145
ing lamellate in old plants, 0.5-1.5 mm. broad; spores white,
smooth, oblong, mostly curved, 1.5-2 x 6.2-7.5 n.
On dead wood or on living trees, especially of Salix. Com-
mon.
This is a very variable species. Sometimes very thin forms
are found and such have been considered as species at different
times. Trametes ruhescens Alb. & Schw. ex Fries is a thin
form with a reddish hymenium. For illustrations, see Hard,
Mushrooms /. 358., White, Hymen. Conn, pi 84. f. 2., and
Moffatt, Higher fungi of the Chicago region yl. 18,
4. D. quercina L. ex Fries, Syst. Myc. i: 333. 1821.
Agaricus quercinus L. Sp. Plant. 1176. 1753.
Plants annual, or sometimes reviving, sessile; pileus dimid-
iate, convex, 4-12 x 4-15 x 1.5-6 cm., corky, whitish to umbri-
nous or almost black, glabrous, margin usually thick and obtuse;
context whitish, corky, 0.2-1 cm. thick; tubes 1-2 cm. long, the
mouths whitish to umber, rarely circular, more often labyrin-
thiform and elongate or lamellate, 1 mm. or more broad, edges
thick and entire.
On Castanea and Quercus, sometimes on the living trees.
Rare.
This species is distinct from all of the others in its habitat,
the thickness of the pileus, and the larger sinuous pores. Hard
(Mushrooms /. 357), and White (Hymen. Conn, pi 34. f.
give illustrations of the plant.
LENZITES Fries, Gen. Hymen. 10. 1836.
Pileus coriaceous to corky, dry and floccose in texture.
Lamella) coriaceous, firm, sometimes simple and unequal,
sometimes anastomosing behind and forming pores; trama floc-
cose and similar to the pileus, the edge subacute. Dimidiate,
sessile, persistent fungi growing on wood and resembling
Dcedalea. (The above description is according to Fries, Epicr.
Syst. Myc. 403.)
This genus is intermediate in position between the Agarica-
ceoB and the Polyporacece and is sometimes included among the
white spored genera of the former family.
[Vol. 1
146 ANNALS OF THE MISSOURI BOTANICAL GARDEN
■y
KEY TO THE SPECIES
r
Context white
Context brown " ■
1 Tubes or interspaces 1 mm . or more broad ; lamella usually not much anas-
3. L. soBviaria
tomosmg ■ ;
1 Tubes or interspaces less than 1 mm. broad; lamellae freely anastomosmg
2.L. vialis
0.3
I. L. betulina L. ex Fries, Epicr. Syst. Myc. 405. 1838.
Agaricus hetulinus L. Sp. Plant. 1176. 1753.
Plant annual, sessile, often imbricate; pileus dimidiate, 2-5
X 2-9 X 0.3-1 cm., coriaceous to somewhat corky, prevailing
color grayish to brownish, marked with many narrow, multi-
colored zones, tomentose; margin thin and acute ; context white,
usually not more than 1 mm. thick; hymenium usually lamellate
but sometimes poroid, the lamellae coriaceous, about 1 mm.
-1 cm. broad, white or whitish; spores globose, smooth,
hyaUne, 5-6 n in diameter.
On all kinds of dead wood of deciduous trees. Common.
L. flaccida Fries as reported from Ohio is but a form of this
species. The plant is well represented by Hard (Mushrooms
/. 185-86), Lloyd (Photogr. pi 14), and fairly well in Cooke,
Illustrations pi. 114^.
2. L. vialis Peck, Ann. Rept. N. Y. State Mus. 26:67.
1874. , ^ ■ ^
Dcedalea pallido-fulva Berk. Hooker's Lond. Jour. Bot.
6:322. 1847.
Pileus annual, sessile, imbricate or single, dimidiate or lat-
erally connate, 1-4 x 2-7 x 0.2-0.8 cm., coriaceous or corky;
grayish brown to cinnamon-brown, often darker at the base,
subtomentose to glabrous, azonate or subzonate, margin acute
context brown, floccose-fibrous to soft-corky, 1-4 mm. thick
hymenium more or less poroid or labyrinthiform, rarely decid
edly lamellate, pores or lamellae averaging 2-3 to a
mm. broad, concolorous with the surface of the pileus; sporei
vcyUndrical to elliptical, smooth, hyaline, 2.7-4 x 7-8.2 n.
On dead wood of both deciduous and coniferous trees. Com
mm.. 1-3
mon
The species was described from Ohio as Dcedalea pallid
fulva Berk, and so reported by Morgan.
\
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 147
3. L. saepiaria Fries, Epicr. Syst. Myc. 407. 1838.
Dwdalea scepiaria Fr. Obs. Myc. i: 105. 1815.
Plants annual, sessile, often imbricate; pileus dimidiate or
reniform, 1-5 x 2-7 x 0.3-1 cm., coriaceous to corky, bright
yellowish red to dark ferruginous, often lighter or discolored with
age, strigose-tomentose, zonate, margin thin; context fulvous
to ferruginous, floccose to soft-corky, not more than 3 mm.
thick ; hymenium usually lamellate, the lamellae about 1 mm.
apart, 2-5 mm. broad, rarely anastomosing, fulvous to rusty
brown; spores cyhndrical, smooth, white, 2.7-4 x 2-10.2 fi.
Always found on dead wood of coniferous trees. Frequent.
Easily distinguished from the preceding species by the deeper
color throughout and by the more distant lamellae that rarely
anastomose.
LinnaDaS:512. 1830.
Plants annual, terrestrial and
ceous, fuscous or cinnamon-colored; context brownish, some-
times rusty brown, floccose to fibrous; hymenium poroid at
first but soon breaking up into concentric lamellae.
The genus is distinct from all others in the concentric arrange-
ment of the lamellae.
C. Greenei Berk. Hooker's Lond. Jour. Bot. 4: 306. 1845.
I.
Pileus stipitate, circular in outline, usually depressed on
top, 2.5-9 cm. broad, 0.5-2 cm. thick, coriaceous when fresh,
rigid when dry, yellowish brown to rusty or purphsh brown,
tomentose at first but becoming glabrous, more or less zonate,
margin thin and acute; context fulvous to cinnamon-brown,
soft floccose to fibrous or somewhat friable, thin at the margin,
thicker next the stipe; tubes 5-8 mm. long, soon breaking up
to form brownish concentric lamellae; stipe central or subcentral,
expanding above into the pileus, velvety, somewhat spongy,
2-7 cm. long, 0.7-2 cm. thick, fulvous to rusty brown in color.
On the ground in woods. Rare.
The species was reported from Ohio by Hard but I think has
not otherwise been collected. For illustration see Hard, Mush-
rooms /. 360-61.
[Vol. 1
148 ANNALS OF THE MISSOURI BOTANICAL GARDEN
FAVOLUS Fries, Elench. Fung, i : 44. 1828.
Plants annual, epixylous, more or less stipitate; pileus fleshy-
tough when fresh, small or medium sized; context white, thin;
tubes in a single layer, the mouths angular, usually hexagonal,
often radiating outward from the stipe and somewhat longer in
the radial direction; spores white.
In our species the stipe is much reduced and is usually lateral
or at least eccentric. The genus is separated from Polyporus
by the large favoloid pores, although some stipitate species of
Polyporus closely approach in pore form the condition ascribed
this family
KEY TO THE SPECIES
Plants about 2 cm. long and broad; hymenium more or less waxy or gelat-
inous iF_ rhipidium
Plants larger than above; hymenium not gelatinous or waxy S. F. canadensis
h
J
I. F. rhipidium Berk. Hooker's Lond. Jour. Bot. 6:319.
1847.
Plants stipitate; pileus reniform, caespitose-imbricate, 2 cm.
long and broad, coriaceous, alutaceous to white, the cuticle
breaking up into minute furfuraceous squamules, concentrically
sulcate; context whitish, thin; tubes short, less than 2 mm. long,
more or less waxy and gelatinous, the mouths white, angular
to elongate, denticulate, averaging 2-3 to a mm. ; stipe lateral,
pruinose, 6-7 mm. long.
On dead wood. Rare.
The above description is adapted from the original. The
species was originally described from Ohio from specimens
collected by Lea. Morgan also probably collected it, but
otherwise it is not known from the state. In habit and color
it resembles Partus sfypticus.
2. F. canadensis Klotzsch, Linnsea 7; 197. 1832.
F. ohiensis Berk. & Mont. Syll. Crypt. 171. 1856. F.
striatulus Ellis & Ev. Am. Nat. 31 : 339. 1856.
Plants stipitate, the stipe often reduced to a lateral tubercle ;
pileus dimidiate to reniform, 1-4x1-8x0.1-0.7 cm., fleshy-
tough when fresh, rigid when dry, at first reddish brown due
to the presence of innate fibrils of that color, later becoming
glabrous and fading to cream color or pure white, azonate,
1914]
OVERHOLTS — THE POLYPORACEiE OF OHIO 149
margin thin and acute, often involute, especially on drying;
context white or whitish, fleshy-tough, becoming firmer on
drying, 0.5-2 mm. thick; tubes 1-5 mm. long, the mouths whit-
ish to yellowish, distinctly angular, usually rhomboid or hex-
agonal, often radiating outward from the stem and longer in the
radial direction, very variable in size, 0.5-3 mm. long and
averaging 1-3 to a mm. in transverse direction; stipe lateral or
rarely subcentral, often rudimentary, not more than 1 cm. long,
1.5-7 mm. thick.
On dead branches of deciduous trees, especially Hicoria.
Common.
F. stnatulus Ellis & Ev. is supposed to differ from F, cana-
densis in having a pileus white in color from the first, and in the
smaller pores. In Ohio both of these forms are found and the
writer has come to the conclusion that F. stnatulus is to be
regarded as only a form of this rather polymorphic species, for
the following reasons: First, specimens of F, canadensis fre-
quently become whitish in color quite early in development;
second, the small pores said to be characteristic of F. striatulus
are also frequently found in specimens with the reddish brown
pileus. In attempting to separate the plants into two species
one finds reddish brown specimens with either large or small
pores, and white specimens with either large or small pores.
The soecies is illustrated in Hard, Mushrooms /. S59.
GLOEOPORUS Mont. Hist. Cuba 385. 1838.
Plants annual, sessile or effused-reflexed; pileus small, thin
and coriaceous; context fibrous, thin, usually white; tubes short,
more or less gelatinous or waxy and in our species separating
from the context in a thin, elastic layer when fresh or when
moistened. The genus is distinct from all others in the gelat-
inous and at the same time separable hymenium. One species
I.
is found in our flora.
G. conchoides Mont. Hist. Cuba pi 15. f. 1. 1838
Sessile or effused-reflexed; pileus dimidiate or conchate,
0.5-3x1-4x0.1-0.5 cm., coriaceous when fresh, rigid when
dry, white or cream-colored, velvety to glabrous, azonate,
margin thin, acute, with a narrow sterile band below; context
white, soft-fibrous, 1-4 mm. thick; tubes less than 1 mm. long,
IVOL. 1
loO ANNALS OF THE MISSOURI BOTANICAL GARDEN
gelatinous or waxy and separating from the context in a thin
elastic layer when fresh or when moistened, the mouths flesh-
colored to reddish purple or purplish black, circular, minute,
averaging 5-6 to a mm.
On dead wood of deciduous trees. Common.
The waxy separating hymenium, reddish purple in color, will
serve to distinguish this species. The plant has been known as
Polyporus dichrous Fries.
MERULIUS
Syst. Myc. i: 326. 1821; HaUer, Hist. Stip. Helv. 3: 150. 1768.
Hymenophore formed from a mycehal mucedinous c
and giving rise to shallow irregular pores formed by the
of obtuse folds of the hvmenium
more
Growing on rotting wood
This genus is a very natural one and forms a transition stage
from the Polyporacece to the Thelephoracece through the genus
Phlebia of the Hydnacece. No special study of the genus has
been made and only the two common species are included here,
although several others have been reported from the state.
KEY TO THE SPECIES
Pileus always present, distinctly pinkish red when fresh 1. M. rubdlus
Pilous when present whitish or somewhat flesh-colored but not distinctly
pinkish red g^ m. Iremellosus
I. M. rubellus Peck, Bot. Gaz. 7: 44. 1882.
Pileus sessile or cffused-reflexed, dimidiate, often imbricate,
3-5 x 5-7.5 X 0.2-0.5 cm., coriaceous-cartilaginous, scarcely
waxy or gelatinous, deep pinkish red, often fading with age,
finely tomentose, azonate, margin thin, acute; context white
or light colored, tough when fresh, soft when dry, 1-4 mm. thick;
tubes short, less than 1 mm. long, formed by anastomosing veins,
averaging 1-2 to a mm., cream-colored or whitish; spores (teste
Peck) minute, elliptical, hyaline 4-5 x 2.5-3 m-
On dead wood of deciduous trees. Common.
This plant is distinguished from the next one by the firmer
consistency and the color, although the color of the pileus often
fades in mature plants. Hard (Mushrooms /. 3S3) gives a
good illustration of the plant.
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO 151
2. M. tremellosus Schrad. ex Fries, Syst. Myc. i : 327.
1821.
M. tremellosus Schrad. Spic. Fl. Ger. 139. 1794.
Sessile, effused-reflexed, or entirely resupinate; pileus dimid-
iate, 0-5 X 3-8 X 0.1-0.3 cm., fleshy or gelatinous-waxy, white
or whitish, tomentose, azonate, margin thin and acute; context
whitish, soft, 1-2 mm. thick; tubes very short, formed by anas-
tomosing ridges or veins, averaging 1-2 to a mm., whitish or
somewhat flesh-colored, in resupinate forms with a wide, thin,
sterile border.
On old logs in woods. Common.
I
Quite often the plant is entirely resupinate and probably
always so in young stages. The form of the hymenium is
exceptionally well shown in Atkinson, Mushrooms /. 191-92.
Besides the above species, M. lacrymans Jacq. ex Fries has
been included in practically every list of fungi reported from
the states east of the Mississippi River, but its frequency of
occurrence is probably in inverse ratio to the number of times
reported. At any rate it is to be considered as a rare fungus
in this country. I have never met with specimens in Ohio that
I could so refer.
mPEX Fries, Elench. Fung, i : 142. 1828.
Hymenium inferior, dentate-lacerate from the first. Teeth
concrete with the pileus, firm, subcoriaceous, acute, reticulately
disposed or arranged in rows, in sessile forms connected at the
base and gill-like, or favoloid in resupinate forms. Basidia
4-spored. Woody, subsessile or resupinate fungi allied to
I
Lenzites and Dcedalea. (Adapted from Fries, Hymen. Eur. 619.)
This genus is sometimes included in the Hydnacece but in at
least one of the three species here described the hymenium is not
toothed from the first, but is decidedly poroid and shows very
close relationships to certain species of the thin pileate members
of the genus Polyporus, e. g., P. biformis, P. proUficans etc., in
which the hymenium soon becomes broken up into teeth. For
this reason and because the plants are very common in our
woods the three following species are described and most of the
collections usually obtained will be found to answer to one of
these descriptions.
[Vol. 1
152 ANNALS OF THE MISSOURI BOTANICAL GARDEN
KEY TO THE SPECIES
Context white or whitish 1
Context brown or brownish 2
1, Context less than 2 mm. thick; tubes or teeth less than 5 mm. long; pileus
villous /. /. tuUpifcra
1. Context more than 2 mm. thick; tubes or teeth more than 5 mm. long. .J?. 7. mollis
2. Hymenium cinnamon-brown 5. 7. cinnanwvieas
2. Hymenium grayish green to olivaceous 4. /. farinaceus
1. I. tulipifera Schw. ex Fries, Epicr. Syst. Myc. 523. 1838.
Boletus tulipifera Schw. Syn. Fung. Car. 99. 1822.
Plants sessile, effused-reflexed, or entirely resupinate; pileus
dimidiate to elongate in outline, 0-1x1-3x0.1-0.6 cm., cori-
aceous, white or whitish, villous, zonate, margin thin and acute;
context white, fibrous, 0.5-2 mm. thick; tubes 1-5 mm. long,
the mouths light colored, averaging 2 to a mm., soon breaking
up into compressed teeth that are connected at the base, and
often with a concentric arrangement.
On dead wood of deciduous trees. Common.
From I. cinnamomeus Fries, and /. farinaceus Fries this
plant is separated by the white or whitish color, and from /.
mollis Berk. & Curt, by the much thinner pileus and the shorter
tubes or teeth.
2. I. mollis Berk. & Curt. Jour. Bot. & Kew Misc. i: 236.
1849.
Pileus sessile or effused-reflexed, dimidiate, 2-5 x 5-10 x 1-3
cm., coriaceous, white or whitish, minutely tomentose to glab-
rous, azonate, margin thin and acute; context white, 2-6 mm.
thick, fibrous; hymenium usually irpiciform, the teeth white,
coriaceous, 0.5-1.5 cm. long, compressed, united at the base.
On dead wood of deciduous trees.
This plant was reported from the Miami valley by Morgan.
I have not collected it in Ohio. It is much thicker than I. tulip-
ifera Schw. ex Fries, and the teeth are much longer.
3. I. cinnamomeus Fries, Epicr. Syst. Myc. 524. 1838.
Pileus none, fungus usually entirely resupinate, coriaceous in
texture, 2-5 mm. thick, entirely cinnamon-brown; context
brown, not more than 1 mm. thick, fibrous; tubes or teeth 1-5
mm. long, becoming toothed at a very early stage, cinnamon-
brown in color, more or less flattened, connected at the base.
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO
153
On dead wood, especially of species of Acer. Rather com-
mon.
Distinguished from the other species here listed by the uni-
form brown color.
4
farinaceus Fries, Linnsea 5: 523
1830
Pileus sessile, effused-reflexed, or resupinate, dimidiate,
0-0.5 X 1-3 X 0.1-0.3 cm., coriaceous, deep brown, finely tomen-
tose, zonate, margin thin and acute; context dark brown,
fibrous, less than 1 mm. thick; tubes 0.5-1.5 mm. long, mouths
usually grayish green or yellowish green, averaging 2-3 to a mm.,
on breaking up into teeth.
On dead wood of deciduous
Not common
Sometimes the fungus is entirely resupinate and then it
usually has a narrow brown margin. It is distinct from all
of the other species in having a greenish hymenium.
INDEX TO THE SPECIES
Names in italics are synonyms, rejected species, etc.
Page Pag
abietinus (Polyporus) 91 chioneus (Polyporus) 97
Abietis (Trametes) 143 cincinnatiis (Polyporus) 114
ahortivus (Polyporus) 105 cinnabarinus (Polyporus) 116
adustus (Polyporus) 102 cinnamomeus (Irpex) 152
albellus (Polyporus) 97 cinnamomeus (Pol3T)oru3) 123
ambigua (Dadalea) 144 circinatus (Polyporus) 121
anax (Polyporus) 113 conchatus (Fomes) 132
applanatus (Fomes) 137 conchatus (Polyporus) 132
applanaius (Polyporus)
t « «
137 conchifer (Polyporus) 93
arcidariformis (Polyporus) 107 conchoides (Glocoporus) 149
arcularius (Polyporus) 107 confragosa (Dacdalea) 144
conglobatu^ {Polyporus) 131
hadiiis (Polyporus) 126 connatua (Fomes) 129
Berkeley! (Polyporus) 113 connatus (Polyporus) 122
betulina (Lenzites) 146 Cratcegi (Lenzites) 144
betulinus (Polyporus) 104 cristatus (Polyporus) Ill
biformis (Polyporus) 95 Curtisii (Polj^jonis) 125
borealis (Polyporus) 100 cuticularia (Polyporus) 118
brumalis (Polyporus) 107
delectans (Polyporus) 99
CECsius (Polyporus) 96 dichrous (Polyporus) 150
canadensis (Favolus) 148 distortus (Polyporus) 105
cameus (Fomes) 131 dryadeus (Pol3T)orus) 119
cameus (Polyporus) 131 drj^ophilus (Polyporus) 120
castanopJiilus (Polyporus) 115 dualis (Polyporus) 121
[Vol. 1
154
ANNALS OF THE MISSOURI BOTANICAL GARDEN
Pa{;e
Page
elegans (PobT)orus) 110 malicola (Trametcs) . 140
endocrocinus (Polyporus) 119 mollis (Irpex) 152
Evcrhartii (Fomes) 134 mollis (Trametcs) 141
molliusculus (Polyporus) 95
farinaceus (Irpex) 153 Morgani {Polyporus) 110
Jihula {Polyporus) 95
fissus {Polyporus) 109 nidulans (Polyporus) 117
flaccida {Lenzites) 146 nigricans (Fomes) 136
flavovirens (Pol\T)orus) Ill nigromarginalus {Coriolus) 93
focicola (Polyporus) 122 nivosns {Trametcs) 143
fomentarius (Fomes) 136
fragrans (PoljTporus) 103 obesus (Polyporus) 121
fraxineus (Fomes) 130 obtusus (Polyporus) 100
fraxinetts (Polyporus) 130 ohiensis (Favolus) 148
fraxinophilus (Fomes) 129 ohiensis (Fomes) 128
fraxirwphilus (Polyjwnis) 129 ohiensis (Trametcs) 128
frondosus (PoljT)orus) 112 ovinus (Polyporus) 126
fulvus (Fomes)" 133
fumosus (Polyporus) 103 pallido-fulva (Dcedalea) 146
palltdus (Polyporus) 109
galactinus (Polj-porus) 98 pargamenus (Polyporus) 92
giganteus (Polyporus) 113 parvulu^ (Polyporus) 122
gilvus (Polyporus) 117 Peckii (Trametcs) 142
gravcolcns (Fomes) 131 pennsylvanicus (Polyporus) 108
Greenei (Cyclomyces) 147 perennis (Polyporus) 122
guttulatus (PohT)orus) 100 pergamenus (Polyporus) 92
per plexus (Polyporus) 118
hirsutulus (Polyporus) 92 piceinus (Polyporus) 143
hirsutus (Pol}T)orus) 93 picipes (Polyporus) 109
hispida (Trametcs) 142 Pilots (Polyporus) 115
hispidus (Polyporus) . 119 Pini (Trametcs) 142
hypococdneus (Polyporus) 115 pinicola (Fomes) 130
pocula (Enslinia) 106
igniarius (Fomes) 135 pocula (Pol}T)oru3) 106
immitus (Polyporus) , 98 pocula (Spharia) 106
intybaceus (Polyporus) 126 popidinus (Fovies) 130
isidioides (Polyporus) 117 poripes (Grifola) Ill
proliferus (Polystictus) 123
lacrymans (Merulius) 151 prolificans (Polyporus) 151
lactea (Tramdes) 144 puhcrula (Polyporus) 103
lacteus (Polyporus) 97 pubescens (Polyporoe) 94
lentus (Polyporus) 126
leucomclas (Polyporus) 126 quercina (Da^dalca) 145
leucophccus (Polyporus) 137
lobatus (Fomes) 137 radiatus (Polyporus) 118
lobatus (Polyporus) 137 radicatus (Polyporus) 110
Lloydii (PoljT)orus) 95 ramosissima (Grifola) 112
lu(;idu8 (PoljTDorus) 123 rcniformis (Polyporus) 137
resinosus (Polyporus) 116
vuiculaius (Polyporus) 100 rhipidium (Favolus) 148
1914]
OVERHOLTS — THE POLYPORACE^ OF OHIO
155
Page
Page
rigida (Trametes) 141 striatdus {Favolus) . . 148
rimosus (Fomes) 133 suaveolens (Trametes) . . 140
roUnice (Pyropolyporus) 133 svbperforatum (Ganoderma) 123
robiniophila (Polyporus) 104 svbsericem {Polyporm) Ui
robiniopMla (Trametes) 104 Svllivantn {Polyporus) 94
roseus {Forms) 131 sulphureus (Polyporus) 114
Rostkomi {Polyporus) 109 Sumstinei {Grifola) 11^
rubellus (Merulius) 150 supinus {Fomes) l^i
rubescens {Trametes) 145
rufescens {Polyporus) 106 tomenlosus {Polyporus) 1^1
tremellosus (Merulius) lol
Bffipiaria (Lenzites) 147 tulipifera (Irpex) 152
salidnus {Fomes) 133
Banguineus (Polyporus) 115 umbellatua (Polyporus) 112
Schweinitzii (Polyporus) 120 unicolor (D^dalea) 14d
scutellatus (Fomes) 128
scutellatus {Polyporus) 128 varius {Polyporus) . 1 10
semipileatus (Polyporus) 96 velutinus {Polyporus) y^
Bepium (Trametes) 139 versicolor (Polyporus) 91
senalis (Polyporm) 139 vialis (Lenzites) 14b
serialis (Trametes) 139 virgineus (Polyporus) 94
sessile (Ganodemm) 123 volvatus (Polyporus) 105
Spraguei (Polyporus) 101
spumeus (Polyporus) 99 zonalis (Polyporus) 101
squamosus (Pol>T)oru8) 109 zorialus (Polyporus) 91
Graduate Laboratory, Missouri Botanical Garden.
Annals
of the
Missouri Botanical Garden
Vol. I
MAY, 1914
No. 2
A CONTRIBUTION TO OUR KNOWLEDGE OF THE
RELATION OF CERTAIN SPECIES OF GRASS-
GREEN ALGiE TO ELEMENTARY NITROGEN
JACOB R. SCHRAMM
Assistant to the Director of the Missouri Botanical Garden
Instructor in the Henry Shaw School of Botany of
Washington University
A general survey of the literature pertaining to the relation
of algse to free atmospheric nitrogen reveals the fact that com-
paratively few forms have been experimented with under
conditions which render the conclusions reached free from
objection. The principal fault which may be found with most
of the work done is that the experiments were carried out with
impure cultures. Representatives from not more than four or
five genera of green alg2e have thus far been studied in pure
culture, and while the general conclusion reached is that these
forms are unable to fix free atmospheric nitrogen either in the
presence or in the absence of combined nitrogen and energy-
furnishing materials, it is by no means certain that forms do not
exist which, under one or all of these conditions, are able to
utilize elementary nitrogen. This thought is especially justified
when the small number of free-nitrogen-fixing species among
the bacteria is considered. In the present investigation, there-
fore, an attempt has been made to extend the observations over
a greater variety of forms in pure culture, — understanding by the
latter a single species of alga free from all other organisms.
Ann. Mo. Bot. Gaso., Vol. 1, 1914
(157)
[Vol. 1
158 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Historical
As early as 1854 Laurent (20^ 21), and Morren (24) occupied
themselves indirectly with the relation of algse to free atmos-
pheric nitrogen. Morren was led to the conclusion that the
sudden death of cultures of infusoria and algse was due to
the insufficient quantity of combined nitrogen furnished when
the number of organisms became considerable. The nitrogen
requirement; he found, could be satisfied by ammonium car-
bonate, organic nitrogenous compounds (decaying insects), and
other nitrogenous substances in the water; but in no case did
he find that free nitrogen from the atmosphere could serve as
the source of nitrogen. While it is difficult to say with what
organisms Morren worked, it is altogether probable that mem-
bers of the Volvocaccce were present among his ''green," "brown,"
and "red infusoria."
No additional contribution to the subject, so far as the author
is aware, was made until the appearance of Frank's paper (9)
in 1888. In his investigation of the question of a possible
fixation of free atmospheric nitrogen in natural soil without the
instrumentality of cultivated plants, Frank exposed samples of
unsterilized soil, poor in organic matter, in containers under a
glass roof, watering them only with distilled water. During
the 134 days that the experiment was continued, no phanero-
gams appeared, but in all cases the surfaces of the soil samples
became covered with a thin, crustlike, greenish layer composed
of "zwei spangriine Oscillariaformen, die eine dick-, die andere
sehr dtinnfadig; ferner griines Chlorococcum humicola, viel-
leicht auch Pleurococcus, sowie Vorkeimfaden von Moosen,
also kryptogame Gewiichse . . . Diatomaceen waren nicht
zu finden." Analysis showed an undoubted increase in total
nitrogen in the experiments. No increase in the nitrate content
was observed, — the additional nitrogen being wholly in the form
of organic nitrogenous compounds. These facts led the author
to the conclusion that the abundance of algal cells, which are
rich in protoplasm and therefore in organic nitrogen, accounts
for the presence of the increased nitrogen in an organic form.
That the appearance of the nitrogen in an organic form (algal
substance) does not represent the primary fixation of free nitro-
1914]
SCHRAMM — GRASS-GREEN ALGiE AND ELEMENTARY NITROGEN 159
gen and that the latter depends on an inorganic process, the inor-
ganic compounds thus produced being subsequently assimilated
by the algae, is not rendered probable by later experiments. In
these, Frank exposed samples of soil, kept free from vegetation,
for long periods of time and at various temperatures. Plant
growth was prevented by leaching the samples daily with hot
water. In this manner any traces of nitrogen compounds
formed were also obtained. Only at high temperatures — too
high for plant growth — did he find a slight increase in total
nitrogen and therefore believes that this process is of no impor-
tance under conditions which admit of plant growth. From
these observations Frank concludes that the algae themselves
are the immediate agents in the fixation of free atmospheric
nitrogen and inclines to extend this faculty to green plants in
general.
In the same year, Gautier and Drouin (11) ascribed an entirely
different function to soil algae. Samples of artificial soils, free
from organic material and containing only ammoniacal nitrogen,
were exposed in a sheltered position for a considerable period of
time. During the progress of the experiments the soil became
more or less covered with a layer of green algae {Pleurococcus
vulgaris, Protococcus viridis, etc.). Analysis showed, in every
even greater loss in ammoniacal
and an intermediate
The
authors assumed that the nitrogen lost was in the form of
ammonia and that the amount of nitrogen appearing in the
organic form was that part of the escaping ammoniacal nitrogen
which, in bathing, so to speak, the algal cells on the surface, was
absorbed, and subsequently built into organic nitrogen
com
pounds. In support of this hypothesis the authors state that
in proportion to the intensity of the algal growth loss in total
nitrogen was diminished, and the amount of ammoniacal nitro-
gen converted into organic nitrogen increased. Gautier and
Drouin thus looked upon the algae as fixers of gaseous ammonia,
which the soil tends to give off constantly, rather than as direct
agents in the fixation of free atmospheric nitrogen.
In 1889, Frank (10) made the fixation of elementary nitrogen
by soil-inhabiting algae the subject of a special investigation.
Four flasks containing sand moistened with distilled water and
[Vol. 1
160 ANNALS OF THE MISSOURI BOTANICAL GARDEN
plugged with cotton were treated as follows : Two were at once
placed in the light; the third was covered with black paper and
without further treatment placed with the first two; the fourth
was exposed for six hours to a temperature of 100°C. and then
placed with the rest. In the first two, rich algal growths de-
veloped; composed of two species of Oscillatoria, a blue-green
" N ostoG-¥ orm , " a yellowish green "Nostoc-Form," a yellowish to
pure green Microcystis, and a Gloeocapsa. In the third and fourth
flasks no growth of any kind developed. Analyses demonstrated
that the total nitrogen content in the first two flasks had been
doubled, whereas that in the latter two had suffered a distinct
loss. The experiments were repeated with unsteriUzed soil^
all air gaining access to the flasks being first passed through
sulphuric acid to remove any ammonia present. The same
characteristic algal flora developed and analysis again showed
a decided increase in total nitrogen. On the basis of these
experiments, Frank makes the generalization that the soil, as
such, is unable to fix free atmospheric nitrogen, and that when
the process does take place, it is effected by means of the vege-
tation of low algse which develop in the soil, and which pos-
sess the ability of assimilating free gaseous nitrogen into vege-
table, nitrogen-containing compounds. He goes still farther and
states that the fact that low algae utilize free nitrogen makes
it more and more probable that the assimilation of elementary
nitrogen is a faculty appertaining to the entire plant world
provided with chlorophyll, and that, since the simple algal cell
is endowed with this faculty, the thought is justified that the
assimilation of free atmospheric nitrogen is as absolute and
fundamental a process of the entire plant kingdom as is the
assimilation of carbon dioxide.
Prantl (27) , in cultivating fern prothallia in solutions with and
without combined nitrogen, observed that whereas an abundant
algal vegetation appeared in the former, only an Anabcena, or
a Nostoc, grew in the latter. When placed in nitrogen -free media,
the blue-green alga always grew abundantly. From this
observation, and without analytical data, Prantl assumes that
free-nitrogen assimilation had taken place, either a direct one
by the alga, or an indirect one in which the alga assimilated the
ammonium nitrite which, according to the theory of Schoenbein,
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 161
is formed in the vaporization of water. Of interest are the
observations by the same author on the unicellular grass-green
algse, which he was unable to cultivate in solutions free from
combined nitrogen. To these^ therefore, he assigned the power
of elementary-nitrogen fixation in a much smaller degree than
to Nostoc,
Frank's conclusions were confirmed by the work of Schloes-
ing and Laurent (31). These investigators supplemented the
usual indirect method of analyzing the soil and harvest, with
the direct method of determining at the beginning and at the
end of the experiment the composition of the atmosphere in
which the plants had been growing. To 2000 or 2500-gram
quantities of a poor sandy soil 2.5 grams of limestone, 5 grams
of a mixture of several rich soils, and a certain volume of a
mineral nutrient solution containing, in some cases, a little
potassium nitrate were added, and the whole placed in large
flasks. In some, seeds of Jerusalem artichoke, oats, peas, and
tobacco were planted; others, to be used as checks, remained
unplanted. To each flask were added 5 cc. of a liquid obtained
by diluting 5 grams of rich soil with 20 cc. of water. After four-
teen weeks, during which time the seeds germinated and pro-
duced plants, the direct analytical method, confirmed by the
results obtained by the indirect method, showed, except in two
checks, an absorption of free atmospheric nitrogen. But the
surfaces of the soils, during the progress of the experiments, be-
came covered with green, cryptogamic plants, among which were
mosses {Bryum, Leptohryum) , and algse {Conferva, scillatoria ,
Nitzschia). This fact led the authors to repeat the first series
of experiments, in every case suppressing the growth of chloro-
phyllous cryptogams by covering the soils with a thin layer of
dry, calcined, quartz sand. No trace of algae or mosses appeared,
and, except in the case of the peas, no absorption of free atmos-
pheric nitrogen was observed. This fact, together with the
evident fixation of nitrogen in the checks of the first series (in
which an abundant chlorophyllous cryptogamic vegetation but
no phanerogamic vegetation developed), and the absence of
fixation in those checks in which little or no algal growth devel-
oped, led Schloesing and Laurent to conclude that there are
some "inferior green plants" which are able to utilize free atmos-
[Vol. 1
162 ANNALS OF THE MISSOURI BOTANICAL GARDEN
pheric nitrogen. In the same year, Gautier and Drouin (12)
reasserted their former conclusion as to the role of algse in nitro-
gen fixation, holding that the methods of those who adhere to
the opinion that algse fix free nitrogen are too faulty to make
conclusions drawn from them convincing.
In the work reported by Schloesing and Laurent in 1892
(32, 33) an attempt was made to reduce the complexity of the
algal cultures by introducing into a single experiment only one
or at most a few species of the algse. All cultures were made on
600-gram quantities of either a subsoil or quartz sand to which
was added (except in the two checks) a small quantity of an
infusion prepared from soils. The cultures were allowed to de-
velop for from three to six months, and, as in the previous experi-
ments of these authors, analyses were made both of the contained
atmosphere and of the soil and algal growth. The chlorophyl-
lous plants which appeared in the various cultures are described
as follows: i and ii — essentially a mixture of Nostoc puncti-
forme Hariot and Nostoc minutum Desmazieres, with a few
colonies of Cylindrospermum majus Kuetz.; iii — almost a pure
culture of Nostoc punctiforme ; iv — Nostoc punctiforme (less
pure than in iii), one colony of Phormidium papyraceum, and
a small quantity of Nostoc minutum; v — two mosses — Brachy-
thecium rutahulum and Barhula muralis; vi — an almost pure
culture of an OscillariecB and Microcoleus vaginatus, with
traces of Tetraspora, Protococcus, Stichococcus , Ulothrix, and
Lynghya; vii and viii — checks with no growths, or at most
a few small patches of Phormidium autumnale Gomont and
Nostoc punctiforme. Both analytical methods showed abun-
dant nitrogen fixation in the first four cultures but not an ap-
preciable one in the fifth, — a fact which the authors explain on
the basis of specific differences in plants in their ability to fix
free atmospheric nitrogen. The checks showed no appreciable
fixation. Separate analyses were made of the top-soil layers,
containing the algal growths, and the deeper layers, the in-
creased nitrogen being found in the algal stratum, — a fact which
the authors consider important in proving that the algae were
responsible for the free-nitrogen fixation. In conclusion, Schloe-
sing and Laurent admit the possibility that the bacteria present
in the cultures had something to do with the fixation of free
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 163
nitrogen, and state that it is not possible to affirm with certainty
that the algae, free from other organisms, are able to effect
fixation. Having observed, however, but few bacteria in the
cultures they conclude that the algse after all are the active
agents in the fixation of elementary nitrogen.
Similar results were obtained by Koch and Kossowitsch (17).
Sixty grams of washed, calcined sand were placed in large Erlen-
meyer flasks and moistened with a mineral nutrient solution
free from combined nitrogen. Since previous experiments had
shown that algae do not grow on sand free from combined nitro-
gen, 0.04 gram of calcium nitrate dissolved in 50 cc. of water
were added to each flask. After inoculation with a suspension
of algal cells obtained from heaps of lime, a continuous slow
stream of air, washed in sulphuric acid, was passed through all
the flasks. Three cultures were placed in a north window, three
in the dark (to determine whether the bacteria contained in
the cultures fixed free nitrogen), and the remainder were
used in determining the initial total nitrogen. After fifteen
weeks, during which time a rich algal vegetation^ developed on
all cultures exposed to the light, the contents of the flasks were
analyzed in toio. Those exposed to the light showed an un-
doubted increase in total nitrogen, whereas those in the dark
showed a slight loss in each case. Of particular interest was one
culture which was brought into the light after it had remained
in the dark for a considerable length of time. After the removal,
a moderate growth of algse appeared, and analysis showed a
slight gain in total nitrogen, which, however, was less than that
found in the cultures which had been exposed to the light during
the entire period. In agreement with the earlier workers, these
authors ascribed to algae the faculty of free-nitrogen fixation,
and emphasized the observation that the extent of this fixation
was directly proportional to the intensity of the algal develop-
ment. Petermann (26) reached a similar conclusion on the basis
of experiments conducted on sterilized and unsterilized soils,
which were respectively inoculated and uninoculated with
algae. The former in each case showed a distinct gain in nitro-
gen, whereas the latter showed either no increase or a slight loss.
> The authors failed to state what algSD developed, merely mentioning the presence
of green and blue-green forms.
[Vol. 1
104 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Incidental to his work on the respiratory quotient in algse,
Schloesing (30) reported that in a culture containing prin-
cipally Protococcus vulgaris Ag., and smaller quantities of
Chlorococcum infusionum Menegh., Ulothrix suhtilis Kiitz., and
Scenedesmus quadricauda Br^b., there was at the end of two
months no diminution of nitrogen in the supernatant atmos-
phere. This fact led the author to place these algae among
those forms which do not fix free atmospheric nitrogen.
As will have been observed, the work reported upon in the
contributions cited was done with impure cultures. A\Tiile in
some cases but a single species was used^ bacteria were present
in all cases. Although in many instances this is not expressly
stated, the author's experience convinces him that the technique
employed by these earlier workers made the contamination of
their cultures with bacteria very probable. It is evident,
therefore, that in the work done thus far it is impossible to
state with certainty whether the results obtained are due to the
activity of the algae, or to the bacteria, or to both.
The first work done on the fixation of free nitrogen by algse
in which pure cultures were used was that of Kossowitsch (18),
in 1894. The only form isolated in pure culture by this investi-
gator w^as one which he states resembled both Cystococcus
(Nageli) and Chlorella vulgaris Bey. He leaves its identity
uncertain but designates it, for convenience, Cystococcus. Pre-
liminary experiments with impure cultures of this alga had
demonstrated that asparagin and ammonium tartrate could
not serve as the source of nitrogen and that growth took place
only when nitrates were supplied. In the experiments with pure
cultures, flasks containing 70 grams of clean sand moistened
with a mineral nutrient solution containing a known amount
of calcium nitrate were inoculated with a carefully tested pure
culture of Cystococcus and allowed to remain four months.
To a number of the cultures dextrose was added, and to others,
in addition to this sugar, pea-tubercle bacteria. At the con-
clusion of the experiments the cultures were carefully tested
for purity. Analysis in every case showed an absence of free-
nitrogen fixation, and demonstrated clearly for the first time
that an alga, Cystococcus, under the conditions reahzed in the
experiment, did not fix free atmospheric nitrogen. That the
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 165
same holds true for this alga in nature seemed probable to
Kossowitsch, who found that it grew vigorously only so long
as a nitrate was present. He further observed that after growth
had ceased in any culture, it was promptly resumed upon the
addition of a nitrate solution, but not when the nitrogen-free
nutrient solution was added. Similar cultures were started in
which the inoculation material was either a mixture of algse
and bacteria derived from soil or Hme, or a mixture of soil bac-
with a pure culture of Cyst
In each case the
with and without dextrose. Table
of these experiments
TABLE I
RESULTS
+ or
Sugar
Content of cultures
Mg. of N in cultures
Initial
Final
+
+
Cystococcus (pure culture)
Cystococcus, Phormidium, soil bacteria,
moulds
2.6
2.6
2.7
2.7
2.6
2.6
7.1
9.5
+
Pure Cystococcus culture and bacteria
2.6
2.6
3.1
8.1
+
+
Stichococcus and bacteria
Nostoc, large round alga, Scenedesmus,
soil bacteria
2.6
2.6
2.G
2.6
2.3
2.7
?
19.1
+
Nostoc, a Cylindrospermwn (small
form), soil bacteria
2.6
2.6
8.8
25.4
Cystococcus, in pure culture, was again unable to fix free
gaseous nitrogen, and the same conclusion is reached by Kosso-
witsch for Stichococcus, which even in the presence of a mixture
of bacteria failed to
elementary
Of
3 the cultures of pure Cystococcus with bacteria, as in
; fixation is ascribable only to the bacteria. Which
of the organisms in the remaining cultures are responsible for
these th
[Vol. 1
166 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the marked fixation of free atmospheric nitrogen it is impossible
to say, the author states. However, from his own results, and
those of previous investigators, that the presence of algae exer-
cises a favorable effect on the process of free-nitrogen fixation,
and, further, that the algae thus far studied in pure culture do
not possess this faculty of fixation, Kossowitsch concludes that
the algae play an indirect role. He believes they do this by
furnishing, through their photosynthetic activity, carbohydrates
to the nitrogen-assimilating bacteria. He would look upon the
algae as occupying the same position with reference to free-living,
nitrogen-fixing bacteria as the legumes do with reference to the
nodule organisms.
Stocklasa (35), while not making his conclusion very clear,
leads one to believe that he considers certain algie (which he
fails to enumerate) capable of fixing free atmospheric nitrogen.
Unfortunately, all of Stocklasa's experiments were carried out
with impure cultures. Molisch (23), in conducting experiments
with algsB relative to the necessary nutrient elements, attempted
to cultivate Microthamnion Kiltzingianum Nag., Stichococcus
hacillans Nag., S. major Rbh., Ulothrix subtilis (?) Kutz., and
Protococcus sp.— all in impure culture— on a nitrogen-free
mineral nutrient solution. In every case the alga) failed to grow,
and Molisch was led to the conclusion that algae require com-
bined nitrogen for their development. Although no experi-
ments in which combined nitrogen was furnished to the algje
were conducted, the author nevertheless makes the statement,
based principally on the work of Kossowitsch just reviewed, that
algae are not able to fix free atmospheric nitrogen.
, In the next year Bouilhac (4) reported that he had succeeded
in isolating in pure culture Schizothrix lardacea, Ulothrix flaccida,
and Nostoc punctiforme. Unfortunately, this author does not
give a detailed account of his isolation methods. Six flasks
containing a mineral nutrient solution free from combined
nitrogen were inoculated with each alga, and to three of each a
drop of soil suspension was added. No growth whatever devel-
oped in any of the Schizothrix and Ulothrix cultures, nor in the
Nostoc cultures to which the suspension had not been added.
But in those cultures of the latter to which a drop of soil suspen-
sion had been added, a splendid growth appeared and in each
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 167
culture analysis showed a nitrogen fixation of from 11 to 23
milligrams. From a second series (in which the cultural solu-
tion contained per liter 0.1 gram arsenic acid in the form of
potassium arsenate) a similar result was obtained, with fixation
of nitrogen of from 5 to 60 milligrams. The presence of Ulo-
ihrix or Pleurococcus in addition to the Nostoc and bacteria
seemed to have no appreciable effect on the quantity of nitrogen
fixed. Bouilhac thus concluded that Schizothrix lardacea and
Ulothrix flaccida (either alone or in the presence of soil bacteria)
and Nostoc punctiforme (in the pure state) are unable to fix free
atmospheric nitrogen in the absence of combined nitrogen.
The abundant fixation in the cultures containing a mixture of
Nostoc and soil bacteria is not ascribed by the author to the
activity of either organism alone.
Richter (28) observed pots of soil with and without plants,
some placed in the dark, others in the light. While a rich algal
vegetation developed in the latter, none appeared in the former.
Only in a few cases was the growth accompanied by a marked
free-nitrogen fixation, but in these instances the author believes
it due to the algge. Pure cultures were not employed. Benecke
(1) contributed some observations made on cultures of Hormi-
diu?n, Vaucheria, Cladophora, and members of the Conjugates,—
all containing bacteria. In nitrogen-free cultures there appeared
what Benecke termed "nitrogen-hunger," a condition which is
characterized in Hormidium by the production of very long, pale
filaments, the cells of which become extremely long and in which
the development of the chloroplast is so meager that the cells
are almost colorless. Stocklasa (36) found that the ''Alinit"
bacteria fix free gaseous nitrogen in much larger quantities
when grown in the presence of species of Stichococcus and Nostoc.
This influence he considers to be due to the pentosans which,
according to his behef, are present in large quantities in various
algse, and which, because of their ready solubility in water,
serve as a favorable energy-furnishing medium for free-nitrogen-
fixing bacteria.
A noteworthy contribution to the subject is that of Kriiger
and Schneidewind (19). These authors for the first time con-
ducted extensive experiments with a variety of algse in pure
culture, including Stichococcus chtoranthus, S. major, S. hacil-
168 ANNALS OF THE MISSOUBI BOTANICAL GARDEN
[Vol. 1
laris, and S. sp., the latter isolated from five different sources ;
Chlorella sp., from tlie group of which Chlorella vulgaris Bey. is
typical (also isolated from five different localities); Chlorella
protothecoides and three other isolations of a form or forms
belonging to the same group; Chlorothccium saccharophihi7n &nd
five other isolations of forms belonging to the same group;
and lastly, Cystococcus humicola. The media employed by
the authors included the following :
1. One per cent dextrose, 0.2 per cent K3PO.J, 0.04 per cent
MgS04, 0.02 per cent CaCl2, and 1 drop of a 2 per cent
FcCU solution to each 100 cc. of solution.
2. Ignited sand moistened with solution 1.
3. Solution 1 plus 0.25 per cent (NHO^SOi, and 0.25 per cent
NaNOs.
4. Ignited sand moistened with solution 3.
5. One-half per cent beef extract, | per cent peptone, and ^ per
cent dextrose.
6. Ignited sand moistened with solution 5.
7. Diluted beerwort.
8. Ignited sand moistened w^ith solution 7.
9. Humolis clay soil plus 35 per cent sand moistened with distilled
water.
The results obtained were uniform in that the media, free
from combined nitrogen, failed to produce a healthy growth,
whereas those containing nitrogen in a combined form showed
an abundant growth,— some of the alga) preferring the nitrogen
in an organic and others in an inorganic form. Further, no fixa-
1 of free atmospheric nitrogen was noted in any of the cultures.
iger and Schneidcwind conclude that there is a strong
probability that all other chlorophyllous soil algai of this kind
are unable to fix free atmospheric nitrogen, and, in general,
agree with the opinion of Kossowitsch that the soil-inhabiting
algae supply the free-living, nitrogen-fixing organisms with the
necessary non-nitrogenous, energy-furnishing material.
Conclusions similar to those of Kossowitsch were reached by
Deherain and Demoussy (8), who succeeded in cultivating blue
lupines free from root nodules in humus-free sand, the surface
of which became covered with Phormidium autumnale and
Uloihrix flaccida in the course of the experiments. The authors
Ki
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 169
explained the growth of the lupines by supposing that the soil
bacteria fixed free nitrogen at the expense of energy-furnishing
organic materials supplied by the algse, and that the nitrogen
so fixed in organic form became available to the legumes.
A return to the conclusion that members of the Cyanophyceoe
fix free atmospheric nitrogen is found in an investigation by
Beyerinck (2). From 1| to 2-liter portions of tap or distilled
water containing 0.02 per cent dipotassium acid phosphate were
inoculated with 1-2 grams of garden soil, and placed in the light.
After several weeks a characteristic growth of blue-green algse
developed, containing, among other species, Anahoena catenula, a
form related to or identical with Nostoc paludosum, and Nostoc
syhcericum , — all non-motile species of Cyanophycece. The devel-
opment of the blue-green algae in an almost nitrogen-free medium
led Beyerinck, without analytical data, and in spite of the evident
contamination of his cultures with soil bacteria, to the con-
clusion that the Cyanophycece belong to the class of organisms
possessing the faculty of free-nitrogen fixation. He regards the
Cyanophycece as the only known organisms capable of synthe-
sizing their organic materials from carbon dioxide and free
nitrogen, and considers as significant in this connection the
observations of Graebner (13) and Treub (37), who found that
in the sequence of floras on fresh sand and lava soils, species of
Cyanophycem are the first to appear.
Cystococcus humicola was once more subjected to a careful
investigation by Charpentier (7). His previous experiments
had demonstrated that the dry weight of algal growth obtained
in liquid glucose media was about one-half that of the weight of
glucose consumed, and that 5.14 per cent of this dry weight
was nitrogen. He then pointed out that the quantity of nitro-
gen furnished by Kossowitsch to his pure cultures of Cystococcus
humicola in the form of potassium nitrate was sufficient to
produce at least 40 milligrams of growth (dry weight) , and that
while this growth was being produced it might not be neces-
sary for the alga to seek nitrogen from the atmosphere. Once
the dextrose was exhausted, the alga might, it is true, develop at
the expense of atmospheric carbon dioxide, but the author holds
the opinion that this would mean a double expenditure of energy
for the assimilation of both carbon dioxide and free nitrogen and
i
170 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
that under these conditions growth would be difficult. Because
of the vast amount of energy necessary for free-nitrogen fixation,
as illustrated by Clostridium P asteurianum , the author suggests
that there is a strong probability that Cystococcus is capable of
assimilating free nitrogen only when the expenditure of energy
in carbon assimilation is reduced to a minimum,— that is to
say, when abundant available organic materials are furnished.
He further emphasizes the necessity of employing combined
nitrogen in a less readily available form than nitrates, suggesting
organic nitrogenous compounds. On media composed of a
decoction of beans to which were added 1 per cent and 2 per cent
of dextrose and gelatin, respectively, Cystococcus was grown
and the entire culture analyzed for total nitrogen. Although
care was taken to have an abundance of available organic
material (dextrose) present, Charpentier found that in no case
was there any indication of free-nitrogen fixation. He further
found that ammonia, asparagin, and peptone w^ere each able to
serve as the sole source of nitrogen.
The association of blue-green algae and soil bacteria is again
referred to as an effective agent in free-nitrogen fixation by
Bouilhac and Giustiniani (5, 6). Buckwheat, white mustard
corn, and cress were planted in clean sand moistened with a
mineral nutrient solution free from combined nitrogen, and the
substrata inoculated with Nostoc punctiforme and Anahcena sp.
covered with bacteria. The phanerogams grew to maturity and
Of
showed a marked fixation of free atmosph
particular interest are the observations of Heinze (14),
who, however, fails to state whether or not the Chlorella ex-
perimented with was in pure culture. He found that no appre-
ciable growth took place in cultural solutions free from combined
nitrogen, but that in the presence of the latter a rich growth ap-
peared, unaccompanied, however, by a definite fixation of nitro-
gen. More important are his experiments with Nostoc in impure
condition, a good growth of the form being obtained in a min-
eral nutrient solution free from combined nitrogen and sugar.
These cultures, as well as others on soil inoculated with a similar
N'ostoc culture contaminated with bacteria and fungi, showed
a definite amount of free-nitrogen fixation. Heinze was unable
to find Azotohacter present, and this, together with the observa-
1914]
SCHRAMM — GRASS-GREEN ALGiE AND ELEMENTARY NITROGEN 171
\
tion that the contaminating fungus in pure culture was unable
to fix free nitrogen, led the author to the conclusion that the
Nostoc iS; in all probability, directly responsible for the free-
nitrogen fixation. Further, he would place Azotohacter in close
relationship with the Chroococcacece, a family in which, he sug-
gests, some forms capable of fixing free atmospheric nitrogen
may be found.
Richter (29), working with pure cultures of Nitzschia palea
and Navicula minuscula, reached the conclusion that the former,
and probably the latter also, is unable to assimilate elementary
nitrogen in the absence of combined nitrogen. Heinze (15),
in experimenting with a Nostoc culture which he had purified
until it contained as a contamination only a Streptothrix , found
that in solutions free from combined nitrogen and sugar but
containing respectively mono, di, and tripotassium phosphate,
a clearly demonstrable amount of free atmospheric nitrogen
was fixed. The Streptothrix was subsequently isolated and tested
as to its ability to fix elementary nitrogen, both with and with-
out sugar, but always with negative results. In conclusion,
Heinze reasserts his former belief that Nostoc is capable of fixing
elementary nitrogen.
Mameli and Polacci (22) succeeded in growing Oedogonium,
Spirogyra, Zygnema, and Protococcus in nutrient solutions free
from combined nitrogen, and demonstrated by analysis an
increase in total nitrogen. They ascribed to these forms, and
to chlorophyllous cells in general, the faculty of synthesizing
ammonia from free nitrogen and nascent hydrogen. Pure cul-
tures were not used. Boresch (3) found that Phormidium
corium. Cohn became brown when grown in solutions containing
very small amounts of combined nitrogen, but that the green
color reappeared following the addition of potassium nitrate
or organic nitrogen compounds. Several species of Oscilla-
ioria, Rivularia, and Chroococcus behaved similarly. But
Anabcena sp. did not change color even when the solution in
which it was growing had become completely exhausted of its
combined nitrogen. While the investigation concerns itself
primarily with the relation of nitrogen to the color in algse,
the observations point once more to species of Anabcena as pos-
sibly belonging to the class of free-nitrogen-fixing organisms, and
172 ANNALS OF THE MISSOUKI BOTANICAL GARDEN
[Vol. 1
equally clearly to the conclusion that the remaining forms
experimented with do not belong to this class.
Oes (25) made the observation that Azolla with its endophytic
Anahcena AzoUcb grew exceedingly well in mineral nutrient solu-
tion free from combined nitrogen. Analysis showed a distinct
fixation of nitrogen. Attempts to cultivate the Anabcena in
pure culture failed. While calling attention to the possible
direct role of the associated bacteria in the observed fixation,
the author inclines to the view that Anabcena Azollce is itself
capable of fixing free atmospheric nitrogen.
The preceding survey of literature shows that in all of the
earlier investigations, and in a considerable number of the later
ones, impure cultures were used. In experiments conducted
under these conditions, it is evident that negative results are, in
general, more reliable than positive ones. Attention should
therefore be called to the negative results which have been ob-
tained from investigations with impure cultures. These, as
will be seen from the literature cited, include a large number of
genera and species from both grass-green and blue-green algae,
and indicate in many cases with a reasonable degree of cer-
tainty that the faculty of elementary-nitrogen fixation is absent
in a very considerable number of species of both the Chloro-
phycece and Cyanophycece. In the former class, all investiga-
tions conducted with pure cultures have led without exception
to the conclusion that these forms are unable to fix free atmos-
pheric nitrogen.
As regards the Cyanophyceos, it should be stated at the outset
that while many observations are on record both affirming and
denying free-nitrogen fixation in the group, it is questionable
whether experiments have been conducted with more than a
single species in pure culture. Bouilhac, it is true, claims to
have isolated Schizothrix lardacea and Nostoc punctiforme in
pure culture. From the meager account given of the isolation
technique, it appears very improbable that the latter form was
actually obtained in culture free from bacteria, although the
former may have been. However, the work of Heinze, while
not conducted with pure cultures, renders free-nitrogen fixation
in Nostoc probable, and it appears especially desirable, there-
r
r
f
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 173
\
fore, to study representatives from this genus as well as other
members of the group Cyanophyceoe.
In the progress of the work about to be reported, it soon be-
came obvious that the development of pure culture methods
would constitute a very considerable portion of the investigation,
and it was deemed advisable to hmit the nitrogen phase of the
problem to the fixation of atmospheric nitrogen in the complete
absence of combined nitrogen, leaving for a subsequent report
the problem of elementary-nitrogen fixation in the presence of
combined nitrogen. The work concerning pure culture methods
will be found reported elsewhere (34).
Experimental
MATERIALS AND APPARATUS
Most of the algsB isolated were soil-inhabiting species. Pre-
liminary experiments showed that in nearly every case better
growth was obtained on sohd media than in liquid ones. For
this reason it was decided to conduct all experiments concerning
the fixation of free nitrogen in the complete absence of combined
nitrogen on a solid medium. Agar could not be sufficiently
freed from all traces of combined nitrogen, and the difficulties
involved in preparing large quantities of silicic acid jelly suffi-
ciently pure were so great that a No. 2| ground quartz was
finally decided upon.
Preparation of the Sand. — The sand, after being thoroughly
washed, was boiled in concentrated hydrochloric acid for two
hours, subsequently washed free from chlorides with distilled
water, and then heated almost to dull redness for from four to
five hours. The sand was then boiled a second time in chemi-
cally pure concentrated hydrochloric acid and again washed
with distilled water until chlorides could no longer be detected.
When this stage had been reached the washing with distilled
water was continued a dozen times more, after which the sand
was drained as thoroughly as possible and the washing completed
with from five to ten changes of nitrogen-free water. After
drying the sand in a clean evaporating dish, a sample was boiled
in nitrogen-free water and the liquid tested for ammonia, nitrites
and nitrates, but only uniformly negative results were obtained.
2
[Vol. 1
174
ANNALS OF THE MISSOURI BOTANICAL GARDEN
Nitrogen-free Water. — The distilling apparatus used was, in
general; like that described by Jones and Mackay (16) for the
preparation of water with a very low electrical conductivity,
except that the water was triply distilled in place of doubly, and
from glass throughout in place of being condensed in a block tin
tube. Fig. 1 represents the distilling apparatus, and it need
only be pointed out that flask in was added to obviate any
possibility of contaminating the distillate with spray from
flask II. The water obtained from this still gave uniformly
negative results when tested for ammonia, nitrites and nitrates.
HHi
R
Fig. 1 . Distilling apparatus for nitrogen-free water
Cultural Apparatus. — One hundred cc. flasks, carefully cleaned
in acid-dichromate cleaning mixture, rinsed in nitrogen-free
water and dried, were connected in series of ten each in the
1912 experiment (eight in the 1913 experiment) by means of
glass tubing and rubber stoppers as shown in pi. 3 fig. 1. The
glass tubing was cleaned in the same manner as the flasks, and
the rubber stoppers were boiled in dilute alkali, then in dilute
hydrochloric acid, and subsequently washed with distilled and
nitrogen-free water. Into each flask of the 1912 experiment an
accurately weighed 40-gram quantity (in the 1913 experiment
1914]
SCHRAMM — GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 175
30 grams) of sand was placed. For purposes of aeration the
separate series of flasks were joined together in groups of five,
as shown in pi. 3 fig. 2, and the free end of the common
connecting tube provided with three sets of triple wash-bulbs,
the two nearest the flasks containing nitrogen-free water,
which served to moisten the air after passing through the third
bulb containing 25 per cent sulphuric acid. In order to aerate
any particular series of flasks it was only necessary to attach
a filter pump to the rubber tube at the end of the series which
it was desired to aerate and to open the pinchcock until the
desired stream of air passed through the wash-bulbs.
Chemicals. — The inorganic compounds used were all Baker and
Adamson's analyzed chemicals; the organic compounds were
Merck's highest purity chemicals.
Cultural Solutions. — In the 1912 experiment, in which each
series contained ten flasks, the following ten cultural solutions
w^ere used and in the following order, the flasks being numbered
correspondingly:
1. NH4NO3 0.5 grams,
MgS04.7H20 0.2 grams,
K2HPO4 0.2 grams,
CaCl2.?H20 0.1 grams,
FeS04 trace,
Nitrogen-free water 1000 grams.
2. The same as No. 1, but containing 0.250 grams of NH4NO3 in-
^
stead of 0.5 grams.
3. The same as No. 1, but containing 0.100 grams of NH4NO3 in-
stead of 0.5 grams.
4. The same as No. 1, but containing 0.050 grams of NH4NO3 in-
stead of 0.5 grams,
5. The same as No. 1, but free from combined nitrogen.
6. The same as No. 5, but containing 2 per cent d-glucose.
7. The same as No. 3, but containing 2 per cent d-glucose.
8. The same as No. 5, but containing 2 per cent mannite.
9. The same as No. 3, but containing 2 per cent mannite.
10. The same as No. 3, but containing 2 per cent saccharose.
In the 1913 experiment, in which each series contained eight
flasks, the following eight cultural solutions were used :
[Vol. 1
176 ANNALS OF THE MISSOURI BOTANICAL GARDEN
!• The same as No. 5 in the 1912 experiment.
2. The same as No. 1 in the 1912 experiment.
3. The same as No- 5 in the 1912 experiment, but with 5.26 grams
of c?-glucose (making a glucose solution isotonic with a 1 per
cent saccharose solution) added.
4. The same as No- 1 in the 1912 experiment; but with 5.26 grams
of (/-glucose (making a glucose solution isotonic with a 1 per
cent saccharose solution) added.
5. The same as No. 5 in the 1912 experiment, but with 5.32 grams
of mannite (making a mannitc solution isotonic with a 1 per
cent saccharose solution) added.
6. The same as No. 1 in the 1912 experiment, but with 5.32 grams
of mannite (making a mannite solution isotonic with a 1 per
cent saccharose solution) added.
7. The same as No. 5 in the 1912 experiment, but with 10.00
grams of saccharose (making a 1 per cent saccharose solution)
added.
8. The same as No. 1 in the 1912 experiment, but with 10.00
grams of saccharose (making a 1 per cent saccharose solution)
added.
The solutions containing the organic compounds were all made
isotonic in order to obviate possible differences in growth due to
different osmotic pressures of the cultural solutions.
The exact volume of solution necessary to just saturate the
amount of sand used in each flask was determined and this
amount of the various solutions added to the corresponding
flasks. The stoppers were then lightly inserted and one group
sterilized at a time in a large Kny-Scheerer horizontal autoclav,
at six pounds pressure for one and one-quarter hours.
INOCULATION
The groups of flasks were transferred directly from the auto-
clav to the inoculating room which had previously been" steamed
down." All inoculations were made with a DeVilbis atomizer,
which, with the excseption of the bulb, is made of metal and
glass throughout. The bulb was removed and the opening of
the metal tip, to which the former is attached, plugged with
cotton. After filling the glass container one-half full of solution
No. 5 (1912 series), the whole (with the exception of the bulb)
1914]
SCHRAMM — GRASS-GEEEN ALGiE AND ELEMENTARY NITROGEN 177
was sterilized. After cooling, the liquid was inoculated with the
desired organism (care being taken to avoid introducing any
agar, which can readily be done if hard agar cultures are used
from which to make the inoculation). The DeVilbis atomizer
is provided with an adjustable metal tip so that the spray may
be directed downward. The metal tip further admits of steril-
ization by flaming. By exercising care and keeping the hands
moist with alcohol, comparatively few contaminations result,
only four having appeared in a total of 320 inoculations.
Attention should be called to the importance of inoculating
in such a way that an approximately equal number of organisms
are introduced and that they are uniformly distributed over the
substratum. Unless this is done growth comparisons cannot
be made with any considerable degree of accuracy, as differences
may be due to localized and unequal inoculation. This is
especially true in algse which do not form motile cells and which,
therefore, are unable to spread rapidly over the substratum.
GROWTH AND OBSERVATIONS
■
All groups of flasks of the 1912 experiment were placed in the
light of north windows at the ordinary room temperature and
the cultures aerated at intervals of from three days to a week.
The 1913 experiment was set up in duplicate, one-half being
placed in a glass incubator kept constantly at from 29.5 to 30.5
C.J and the other half in a similar incubator at the ordinary room
temperature. Both series of cultures were placed directly in
front of a north window and were aerated from time to time.
Space will not permit the detailed tabulation of the observa-
tions on growth. In the following tables, growth is indicated
without reference to time. A few general statements may,
however, serve to give some idea as to the relation of the com-
position of the cultural medium to the time elapsing before a
macroscopically visible growth appeared. In almost every
case, growth was observed first on the glucose-containing medium
and almost as soon or slightly later on the one containing sac-
charose. It should be said, however, that a healthy growth
w^as maintained on these two media, in most cases, for but a
short time. Chlamydomonas pisiformis Dill forma minor
Spargo is a marked exception in this respect, a splendid, healthy
178
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
TABLE II
RESULTS OF THE 1912 EXPERIMENT
(August 6, 1911— July
1, 1912.)
Organism
Sol. 1*
(with
comb.
nitrogen)
Sol 5
(with-
out
c.n.)
Sol. 6
(with-
out
c.n.)
1
Sol. 7
(with
c.n.)
■
Sol. 8
(with-
out
c.n.)
1
Sol. 9
(with
c.n.)
Sol. 10
(with
c.n.)
Chlamydomonas pisifor^
mis Dill foima minor
Spargo
-t
++++
++++++++
Chlorella sp., large form
with clathrato chro-
matophore
+ + +
^
++
1
1
1
—
+++
+ +
Kirchneriella sp.jO, form
without marked gela-
tinous envelope
+
P
—
^
1
1
1
1
1
+
1
1
—
+
+
Protosiphon botryoides
(Kiitz.) Klebs
-t
-t
1
+++
1
1
—
+++
+++
Chlorococc urn hu micola
(Nag.) Rabenh;
+ + +
_
++++
^
+++
++++
Chlorella vulgaris Bey.
+ + +
-t
_
++
—
+
++
Stichococciis bacillaris
Nag.
F
+ + +
—
_
++
1
1
Slight, fair, good, and splendid growths are respectively indicated by +, ++,
+ + +, and + + + + . No growth is indicated by — .
The mention of growth in solutions 2, 3, and 4 is omitted because growth dif-
ferences were not marked. These solutions were introduced in each series in order
r
to note the effect of steadily decreasing quantities of ammonium nitrate on the in-
tensity of growth.
t A scarcely detectable growth developed in these cases which, however, disap-
peared in all cases within a short time. The growth was so slight as to be notice-
able only when the flasks were compared with others absolutely free from growth.
growth being maintained for a very long time. Growth on the
mannite-containing medium was usually slower in making its
appearance, but, in general, remained healthy for a longer
period of time than those on glucose or saccharose. With
very few exceptions, growth appeared last on the purely syn-
thetic medium, but was maintained in a state of vigor longer
1914]
SCHRAMM — GRASS-GREEN ALGJE AND ELEMENTARY NITROGEN 179
TABLE III
RESULTS OF THE 1913 EXPERIMENT
(March 29-April 16, 1913.)
Organism
Sol. 1
(with-
out
c.n.)
Sol. 2
(with
c.n.)
Sol. 3
(with-
out
c.n.)
Sol. 4
(with
c.n.)
Sol. 5
(with-
out
c.n.)
Sol. 6
(with
c.n.)
Sol. 7
(with-
out
c.n.)
Sol. 8
(with
c.n.)
Chlamydomonas pisi-
formis Dill forma
minor Spargo. '
Temp. 18.5-24°C.
++
+++ -*
++
+ + +
Ditto. Temp. 29.5-
30.5"C.
_
++
1
i
1
1
—
+ +
Chlorella sp., large
form with clath-
rate chromato-
phore. Temp. 18.5
-24°C.
1
+
1
1
+
1
i
]
1
1
1
1
+ + +
Ditto. Temp. 29.5-
30.5°C.
—
+
- + +
1
1
1
+
+ +
Stichococcus bacillaris
Nag. Temp. 18.5-
24 °C.
1
+
'
^^ . .
1
++
L
+ +
Ditto. Temp. 29.5-
30.5°C.
1
1
—
1
++
1
— 1
_-
+ +
Chlorococcum humi-
cola (Nag.) Ra-
benh. Temp. 18.5
-24°C.
+
1
i
]
1
1
1
1
+
1
■
1
1
1
+ + + +
Ditto. Temp. 29.5-
30.5°C.
1
1
M
+++
- 1 -
+ + +
Protosivhon hotryoi-
des (Kiitz.) Kleba.
Temp. 18.5-24°C.
1
+
+++
1
1
+ +
1
+ + +
Ditto. Temp. 29.5-
30.5°C.
—
+++
"
—
+ + +
Chlorella wlgarisBey.
Temp. 18.&-24°C.
+
++
+
+ +
Ditto. Temp. 29.5-
30.5°C.
_
^_^^.^,^_^^_
—
^
Kirchneriella sp.
Temp. 1S.5-24°C.
—
—
—
-^
Ditto. Temp. 29.5-
30.5°C.
1
1
—
H
1
_
A scarcely detectable and rapidly disappearing growth developed as in the
1912 experiment.
180 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
[Vol. 1
than on any of the organic-compound-containing media. Glu-
cose, saccharose, and mannite were chosen as energy -furnishing
compounds because of their general usefulness in this capacity
among free-nitrogen-fixing bacteria, and also because they are
representatives from three great classes of carbon compounds.
Certain unpublished experiments, carried out by B. M. Dug-
gar on nitrogen fixation in the fungi, indicate that, whereas no
fixation takes place at ordinary temperatures, it does take place
at elevated temperatures. It was thought desirable, therefore,
to investigate the effect of elevated temperature on the process
of elementary-nitrogen fixation by algse in the absence of
combined nitrogen. However, the results tabulated in table
III show clearly that not only did no growth on any nitro-
gen-free medium appear at the higher temperature, but also
that that appearing on nitrogen-containing media was, in many
cases, poorer than that obtained in cultures kept at ordinary
temperatures. It should further be noted that growth was in
some cases entirely suppressed. It would appear, therefore, that
in the species investigated, growth at elevated temperatures is
less vigorous than at ordinary temperatures and that, in all
probability, no favorable effect on free-nitrogen fixation is to be
expected by growing these species at the higher temperature
maintained in the experiment.
The incipient, ephemeral growth which was observed in a
few cases where combined nitrogen was not furnished is be-
lieved to be due to the minute quantity of combined nitrogen
which was unavoidably introduced in the inoculation process.
The inoculating material was, of necessity, derived from agar
containing ammonium nitrate, and while no agar was trans-
ferred it is altogether probable that enough combined nitrogen
was carried over in the water adhering to the cells to account for
the trace of growth. It should be emphasized again that in
every case this growth was so shght as to have escaped detec-
tion had not a comparison been made with a flask absolutely
free from growth.
In table iv the results of the two experiments are combined
and show that in seven species complete results have been ob-
tained. These results indicate with perfect uniformity that
growth, under the conditions realized in the experiments, is
impossible in the absence of combined nitrogen, even when readily
1914]
SCHRAMM— GRASS-GREEN ALG^ AND ELEMENTARY NITROGEN 181
TABLE IV
SUMMATION TABLE OF 1912 AND 1913 EXPERIMENTS
(Solutions numbered
as in 1913
experiment.)
Organism
!
1
1
Sol. 1
1
(with-
out
c.n.)
Sol. 2
(with
c.n.)
Sol. 3
(with-
out
c.n.)
Sol. 4
(with
c.n.)
1
1
Sol. 5
(with-
out
c.n.)
Sol. 6
(with
c.n.)
Sol. 7
(with-
out
c.n.)
Sol. 8
(with
c.n.)
Chlamydomonas
pisiformis Dill
forma minor
Sparge
i
1
+ +
1
++++
1
1
+ + + +
1
1
+ + + -I-
Chlorella sp.
—
+++
—
++
-
- +++
+ + +
Stichococcus bacil-
laris Nag.
—
+++
++
-^
+
—
+ +
Chlorococcum hu-
micola (Nag.)
Rabenh.
—
+++
1
++++
—
+ + +
^^■^^
+ + + +
Protosiphon botry-
oides (Kiitz.)
Klebs
1
1
++
!
+++
1
J
+ + +
—
+ + +
Chlorella vulgaris
Bey.
+++
1
++
+
, ^^
r
+ +
Kirchneridla sp.
_
+
+
—
+
-^
+
assimilable energy-furnishing compounds like glucose, mannite
and saccharose are suppHed; and that, therefore, these forms
under the conditions stated, are totally unable to fix free atmos-
nheric nitroeen in the complete absence of combined nitrogen.
Conclusions
1, In agreement with all work that has previously been done
on the assimilation of elementary nitrogen by grass-green algae
in pure culture, it has been found that Chlamydomonas pisiformis
Dill forma minor Spargo, Protosiphon hotryoides (Kiitz.) Klebs,
Chlorococcum humicola (Nag.) Rabenh., Chlorella vulgaris Bey.,
Stichococcus hacillaris Nag., Chlorella sp., and Kirchneriella sp.,
are unable to fix free atmosph
nitrogen in the com
182 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
absence of combined nitrogen, under the conditions realized in
the experiments.
2. A sHghtly elevated temperature (from 5 to 10° C. above the
ordinary range of room temperature— 18-24 °C.) does not, as
is the case in certain fungi, enable the algse investigated to fix
free gaseous nitrogen in the complete absence of combined
nitrogen.
In conclusion, the author wishes to express his sincere apprecia-
tion and gratitude to Dr. George T. Moore, at whose suggestion
the work reported upon in this paper was undertaken and under
whose constant attention and generous aid it was carried to
completion; to Dr. B. M. Duggar, for many valuable suggestions
and innumerable courtesies; to Mildred Spargo Schramm, for
kindly encouragement and help throughout the investigation;
and to Dr. George R. Hill, Jr., for substantial aid during the
of the work
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Jahrb. 35:889-910. 1906.
16. Jones and Mackay, A contribution to the study of water solutions of some of
the elemcnta. Am. Chem. Jour. 19: 83-117. 1897.
17. Koch, A., und Kossowitsch, P. Ueber die Assimilation von freiem Stickstoff
durch Algen. Bot. Zeit. 51 : 321-25. 1893.
18. Kossowitsch, P. Untersuchungen iiber die Frage, ob die Algen freien Stick-
stoff fixiren. Bot. Zeit. 52:97-116. 1894.
19. Kriiger, W., und Schneidewind, W. Sind niedere chlorophyllgnine Algen im-
stande den freien Stickstoff der Atmosphare zu assimilieren und den Boden an
Stickstoff zu bereichem? Landw. Jahrb. 29 : 776-804. 1900.
20. Laurent, M. P. Recherches physiologiques sur lea animalcules des infusions
vegctales compar^es aux organes 61^mentaires des v^g^taux. Pans. 1854.
21. , Recherches sur les Infusoires. Reclamation de priority addressee k
I'occasion d'une communication de M. Morren. Compt. rend. acad. Paria
39: 1034. 1854.
22. Mameli, E., and Polacci, G. Su I'asimilazione diretta dell'azoto atmosferico
libero nci vegetah. Atti Istit. Bot. Pavia 15: 159-257. 1911.
23. Molisch, H. Die Emahrung der Algen (Susswasseralgen : I Abhandl). Sitzungs-
ber, d. k. Akad. d. Wiss., Wien, math.-naturw. Kl. 104: 783-800. 1895.
24. Morren, M. De I'absorption de I'azote par les animalcules et le3_ alguea.
Compt. rend. acad. Paris 38:932-34. 1854.
25. Oes, Adolph. tjber die Assimilation des freien Stickstoffs durch AzoUa. Zeit-
schr. f. Bot. 5:145-63. 1913.
26. Petermann, A. Contribution h la question de I'azote. Bull. Acad. Belg. III.
25:267-76. 1893.
27. Prantl, K. Die Assimilation freien Stickstoffs und der Parasitismus von Nostoc.
28. Richter, 0.
135-36. 1889.
:;ar Frage der Stickatoffernahrung der Kulturpflanzen. Landw.
221-41. 1898,
29. , Zur Physiologie der Diatomeen. Sitzungsber. d. k. Akad. d. Wias
Wien
1906
30. Schloesing, Th. Sur les ^changes d'acide carbonique et d'oxygSne entre les plantes
et ratmosphSre. Compt. rend. acad. Paris 117: 813-16. 1893.
31. Schloesing, '1 h. fils, et Laurent, Em. Sur la fixation de I'azote libre par lea
plantes. Compt. rend. acad. Paris 113 1 776-79. 1891.
32. , — , Sur la fixation de I'azote libre par les plantes. Compt.
rend. acad. Paris 115:659-61. 1892.
33. ^ , Recherches sur la fixation de I'azote libre par les plantes.
" Ann. Inst. Pasteur 6: 65-115, 824-40. 1892.
34. Schramm, J. R. Some pure culture methods in the algae. Ann. Mo. Bot. Gard.
1 : 23-45. 1914.
35. St'ocklasa, J. Studien uber die Assimilation elementaren Stickstoffs durch die
Pflanzen. Landw. Jahrb. 24: 827-63. 1895.
36. , Assimilieren die Alinitbakterien den Luftstickstoff? Centralbl. f.
Bakt. II.' 5: 3.50-54. 1899;6:22-24. 1900.
37. Treub, M. Notice sur la nouvelle flore de Krakatau. Ann. Jard. Buitenzorg 7:
213-23, 1888.
. Q [Vol. 1, 1914 ]
I04 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
Fig. 1.
arrangement
PLATE 3
; quartz s£
_ liQ 2 Five series of culture flasks arranged in a group with a common connect-
ing tube (on the left) and a series of three triple wash-bulbs. On the right, the rubber
tubing, provided with pinchcocks, is shown attached to each series for use in aeration
Ann. Mo. Box. Gard.,Vol. 1. 1914
Plate 3
Fl(i 1
SCHRAMM
GREEN ALGAE AND ELEMENTARY NITROGEN
toe K A Y N E . HOST( ) N .
THE THELEPHORACEJE OF NORTH AMERICA. I
EDWARD ANGUS BURT
Mycologist and Librarian to the Missouri Botanical Garden
Associate Professor in the Henry Shatv School of Botany of
Washington University
1
Inteoduction
This monographic study of the North American Thelephoracem
was begun in 1894 as the author's contribution towards a greatly-
needed manual of the Basidiomycetes of the United States, — a
need that still confronts us. It has been necessary to carry on
these investigations in connection with college and other work
which required most of my time, but the long peripd covered
has been an advantage; for during these two decades there has
been such widespread interest in the Thelephoraceoe on the part
of American students of fungi that it has been possible to study
this family and its distribution from extensive series of freshly
collected specimens from all the important regions of North
America with the exception of Alaska, Mexico, and the Colorado-
New Mexico region of the United States, from which but
small collections have been received. These specimens have
been preserved unpoisoned in my herbarium in insect-proof
tin boxes which receive herbarium sheets, and each will be
cited by the number or other designation adopted by my cor-
respondents in order that their specimens may be as useful for
future reference as my own. The quantity of material always
awaiting examination has confined my work to a systematic
treatment of this family.
Except in the case of types of species, specimens of published
exsiccati, and the specimens of Schweinitz's herbarium, I cite
but few specimens from the large herbaria. This is done on
account of the difficulty and large amount of time involved in
making a study of the material contained in them. Serious
changes in the condition of the specimens in these herbaria have
been occasioned partly by time but more largely by the poison-
ous solutions with which the specimens were soaked for preser-
vation under old-fashioned methods of herbarium procedure;
Issued July 1, 1914.
Akn. Mo. Bot. Gabd., Vol. 1, 1914
(185)
186 ANNALS or THE MISSOURI BOTANICAL GARDEN
[Vol. 1
methods well enough adapted for flowering plants but not for
fungi.
Early in the w^ork it became apparent that the diagnoses of
known species of resupinate Thelephoraceoe had failed utterly to
enable the leading working mycologists of any country to recog-
nize with certainty in the species about them those described in
other countries, or those described for their own country by
earlier students. The truth of this statement is shown by the
errors and confusion in names of the common species which
have been distributed in exsiccati, by the fact that in the large
herbaria several different species are Ukely to bear the same
specific name on the same or successive sheets, and by the
vastly more important fact that the masters of mycology of each
age, when relying wholly on the diagnoses published by their
contemporaries or predecessors, have described as new species
common and conspicuous resupinate fungi which had been
accurately described by immediate contemporaries or prede-
cessors, and in very many cases just as accurately by still earlier
students. All the mycologists concerned in these redescriptions
have been
truth, I am convinced, and
would have preferred to employ the earlier names for their plants,
could they have known that those earlier names referred with
certainty to their specimens. All these people were relying,
as was the usage of their time, on a few words of published
description in some other than their mother tongue.
It is time to recognize generally that the resupinate i77/7?2e72o-
mycetes, and especially the Thelephoraccce, are extremely diffi-
cult taxonomic problems. Descriptions must include more
than a rather vague and generalized characterization of the
mere superficial appearance and habit of the specimen with
possibly a reference to spores which some one recorded for what
was perhaps this species. The fungus itself is an individual of
the species; the description in words and by illustration has
merit in proportion to the success it has in producing in the
mind of any educated stranger exactly the ideas which he could
derive from the study in detail of the specimen itself. From
the specimen, exact ideas may be had of coloration, of form,
of dimensions, of texture, of consistency, of internal structure,
of organs of minute size, of place of growth, and of host and
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA, I 187
m
substratum. If the description fails to give the color as exactly
as if it had been noted by comparison with such a standard
work as Ridgway's 'Color Standards' or Saccardo's 'Chromo-
taxia/ then it is inferior to the specimen; if the description con-
tains no information as to whether the basidia are simple or
cruciate, making up the whole hymenium or arranged side by
side with other organs of characteristic form, standing directly
on the substratum or separated from it by densely or loosely
interwoven hyphse or other form of subhymenial layer; — if it
does not contain all this information in exact terms and as much
in addition as the specimen itself could afford, then it is an im-
perfect description of the species. It may be so imperfect that
a dozen different species of fungi could be assembled, to any
one of which it would apply as well as to any other, as is the
case with the supposedly common and cosmopolitan Corticium
lacteum and C. calceum. Published exsiccati probably contain
the full dozen under each of these names.
In the case of resupinate Hymenomycetes, types and authentic
specimens of the species are of the highest importance to supple-
ment the prevailingly imperfect descriptions with full and
exact data. Hence, the types of fungi on which the descriptions
are based and the authentic specimens from the authors of the
species are of importance in proportion to the degree in which
these plants may yield data not afforded by the descriptions and
existing illustrations of the species. In the case of the resupi-
nate Hymenomycetes, the early descriptions are of slight prac-
tical value except as they are backed up by types and specimens
from their authors. For this reason, if there had been no
other, the International Botanical Congress, at Brussels, acted
for the best interests of mycology in fixing the beginning of the
naming of Hymenomycetes with the publication of Fries* 'Sys-
tema Mycologicum/ — the time when the preservation of types
and authentic specimens of such fungi in herbaria became so
prevalent that it was possible for later mycologists to distin-
guish the resupinate species by taking the trouble to study the
types, if authentic specimens could not be obtained.
My method of becoming acquainted with our described
species of Thelephoracece has been to study and arrange by
species in my herbarium the specimens as they have accumu-
188 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
lated. In this arrangement due regard has been given to origin
nal descriptions of species and to all details of internal structure.
Spore collections on glass shdes have been made for each species
whenever possible, and about five thousand mounts of sectional
preparations in glycerin have been made from collections and
preserved for reference in connection with internal structure of
the specimens. From time to time I have taken my Thele-
phoracecE to herbaria where the types of our American species
are stored and have there painstakingly matched them with
the types. I have made sectional preparations from a frag-
ment of each of these types in order to make sure that my
specimens match the types not only in external characters but
also in all details of internal structure. The sectional prepara-
tions of type specimens have been preserved in glycerin. Speci-
mens from my herbarium which have been so matched with type
specimens have been used by me later for the determinations
of subsequent collections. Such methods of investigation are
probably too laborious and require too much time to become
popular and they afford little opportunity for the inspirational
flights attributed to genius, but they do afford a means of deter-
mining within very narrow limits the species of North American
Thelephoracece.
I am under especial obligation to Dr. W. G. Farlow for sug-
gesting this work, for interest in its progress, and for frequent
access to the Curtis Herbarium for comparisons with types. I
am indebted also to Dr. C. H. Peck for opportunity to study
his types in the New York State Herbarium, to the late Dr.
L. M. Underwood for similar opportunity with the Ellis types
in the Herbarium of the New York Botanical Garden, to Dr.
S. W. Dixon and Professor S. Brown, of the Philadelphia Acad-
emy of Natural Sciences, for the privilege of studying in the
Schweinitz Herbarium, to Sir W. T. Thistleton-Dyer and Mr.
G. Massee for access to types and authentic specimens in Kew
Herbarium, to the late Dr. T. M. Fries for the privilege of
studying in the Herbarium of Elias Fries, at Upsala, and to Mr.
Lars Romell, of Stockholm, Dr. P. A. Karsten, of Mustiala,
and Abate G. Bresadola, of Trient, for many authentic speci-
mens of their own species and for specimens which they had
compared with types of early authors of Thelephoracece of
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 189
Europe. In the later pages names of the many botanists who
have participated in this work by the contribution of speci-
mens from their respective regions are given in connection
with the specimens. I feel my obhgation to each of these cor-
respondents.
Having become thorouglily familiar with the species of a
family of fungi, one then faces the task of deciding under what
genera they shall be grouped in order that others may more
easily recognize them. Our studies in systematic botany and
the accumulations of plants in herbaria are primarily for the
purpose of enabling those who wish to obtain information about
any particular plant, however obscure, to determine its name
accurately and so be in a position to get at the world's literature
and knowledge concerning that species; and also to enable
botanists so to entitle and index their researches that the
results will be more available to the world at large. Stability
in the nomenclature of plants is therefore important, and revo-
lutionary changes in generic conceptions should not be lightly
and frequently made. Whenever one proposes new genera to
supersede a well-established genus which has satisfactorily
embraced the related species of the world, the burden of proof
should be on the one who makes the change to demonstrate
that the advantages from the innovation will more than com-
pensate for the confusion which would result as well as for the
loss of knowledge indexed under the superseded name.
Many new genera of fungi have been proposed during recent
years. These have frequently come from students with a
limited knowledge of the species of the world. It is not sur-
prising that a botanist working on the few species of a limited
region should be led to the establishment of new genera on the
basis of what seem to be sharp differences in his species or groups
of species. When, however, his knowledge encompasses just
as definitely the structure of the many species of some large
portion of the world, his perspective changes, and he may now
find that the species which he formerly regarded as generically
distinct are so closely connected by intermediate species that
the contemplated generic separation would be unnatural and a
hindrance to botanical progress. It is fundamental that genera
be so sharply defined that any accurate observer who will make
3
[Vol. 1
190 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the study necessary for the application of the generic definition
may be sure ninety-nine times out of a hundred that the fungus
on which he is working is a Stereum, for example, and not a
Thelephora, nor a Craterellus, nor a Cladoderris, nor a Corticium,
nor a Peniophora, nor a Sebacina. It is an obhgation on authors
to group their species so accurately under genera that Stereum,
for example, shall comprise all the species of this genus known to
science, and no others. The synonomy of species in later pages
will show how vaguely the genera of Thelcphoracece have been
comprehended.
It is desirable that a genus should consist of but few species
in those cases where the group is sharply and naturally set off
from others, that is, where no intermediate species connect the
genus with other groups. While such small genera are desir-
able, if wholly natural, it is in the highest degree objectionable to
create small artificial genera by arbitrarily segregating the
species of a natural genus and so establishing indefinite fines
of demarkation between genera. Under such a procedure the
generic location of certain species becomes wholly arbitrary
and always continues as a stumbfing block for new students
and this leads to the loading of our literature with so-called new
species. A case in point is Saccardo's scheme in the ^Sylloge
Fungorum' in which he separates Hypochnus from Corticium
and Peniophora without any natural generic planes of cleavage.
In practical work one needs to know exactly what the generic
limits of Corticium, Peniophora, and Hypochnus are. The
question naturally arises as to just how loose and open the
structure of the fructification must be to be included in the
genus Hypochnus rather than in Corticium or Peniophora.
Henning's violation of the principle involved is still more fla-
grant, for he separated the Hypoch7iacea as a new family from
the Thelcphoracece^ and placed Hypochnus of Saccardo in the
Hypochnacece, and Corticium and Peniophora in the Thelephora-
cece. As all students of the Thelcphoracece have found Hypoch-
nus, as understood by Saccardo, wholly unworkable, it would
increase the usefulness of the 'Sylloge Fungorum' if Saccardo were
to distribute among Corticium and Peniophora, the species
which he now includes under Hypochnus.
lEnglcr und Prantl, Nat. Pflanzenfam. (I. 1**): 114. 1898.
1914]
BUKT — THELEPHORACE.E OF NORTH AMERICA. I 191
Probably all species of Corticium, as originally understood,
have an hymenium composed of basidia arranged side by side
between non-sporebearing organs termed paraphyses. In many
species, it is difficult to distinguish between the basidia and
the paraphyses except by prolonged study of special prepa-
rations or by observations made at the time the basidia bear
spores. In other species the sterile organs are conspicuous and
distinct from the basidia either by their larger size, different
form, or thicker or incrusted walls. Such conspicuous bodies
are called cystidia, but if the paraphyses are merely finely but
characteristically branched near their tips, they are not called
cystidia. Such branched paraphyses occur in the hymenium
of occasional species of several genera of the Thelephoracece and
are valuable characters for specific diagnosis.
In 1880, Cooke proposed, from Kew Herbarium, to divide
the old genus Corticium into two genera, — the name Corticium
to be retained for those species having the non-sporebearing
organs of the hymenium not distinguishable from the basidia,
and the generic name Peniophora to be given to those species
having cystidia. As the species of Corticium were very num-
erous and extremely difficult taxonomically, this proposal was
hopefully received, and for more than thirty years the transfer
of species from Corticium to Peniophora has been going on and
the end has not been reached yet. During this long period
there has been confusion as to which species of the old genus
Corticium belong in the emended Corticium and which in the
genus Peniophora.
Peniophora is an artificial rather than a natural genus, how-
ever, and its adoption has given to many species a position
intermediate between this genus and Corticium. These inter-
mediate species have to be classed with the one genus or the
other according to personal judgment, for no one can state just
how conspicuous the sterile organs must be, nor of how constant
occurrence, to merit the name cystidia. In Corticium Samhuci
Fr., for example, cystidia are readily found in preparations from
some collections, but several preparations may have to be made
to demonstrate them in other collections. In the same species ,
and in different parts of the same section, cystidia may some-
times be sparingly and sometimes not at all incrusted. Some
[Vol. 1
192 ANNALS OF THE MISSOURI BOTANICAL GARDEN
species which I have placed in the genus Peniophora because
of the presence of cystidia students may look for under Cortidum
when, by a more hasty study of their collections, they fail to
detect these organs. On the other hand, students using more
discriminating methods than mine may detect cystidia in species
in which I have overlooked them, and such students will search
in Peniophora for species which I have placed under Corticiwn.
Species intermediate between genera always cause such trouble.
There are many intermediates between Peniophora and Cortic-
ium, yet in this particular case the advantage from the separa-
tion undoubtedly more than compensates for the disadvantages
occasioned by the intermediate species.
The case of Peniophora has been considered at length, be-
cause this genus is being regarded as a precedent for subdividing
Stereum and grouping under Lloydella all those species which
have conspicuous non-sporebearing organs between the basidia.
Such a separation, however, would be artificial and give rise to
a troublesome series of intermediate species, without the com-
pensating advantage which accrued in the case of Peniophora
and Corticium. Stereum is not a genus of difficult species nor does
it comprise an immense number of species. It is just a fine, nat-
ural group of species capable of being more sharply defined
than it was by Fries, so as to receive some species from The-
lephora of Fries and to part with some to Corticium, So defined,
even beginners will have no trouble in recognizing species of
Stereum. Systematic work in mycology should strive to estab-
lish and maintain just such natural, clean-cut genera as Stereum.
It seems to me best to work along constructive rather than
destructive lines. Fries had a wonderful ability for the per-
ception of the natural grouping of fungi on the basis of gross
morphology and habit. Since his time, research has greatly
enlarged the knowledge of the internal structure of fungi and of
the organs of propagation. The value of such organs in the
classification of seed plants is well known. It is feasible to
modify somewhat the genera of ThelephoracecB as defined by
Fries, in accordance with the true relationships and differences
shown by the present knowledge of internal structure, basidia,
and spores, and a system results which is the natural evolution
of taxonomic and morphologic study of Thelephoraccce. This
1914J
BURT — THELEPHORACE^ OF NORTH AMERICA. I 193
system has been communicated to my correspondents in con-
nection with specimens. Its principal features are:
1. To restrict Thelephora to pileate species with simple basi-
dia and colored spores.
2. To follow Karsten and Bresadola in placing under Hypoch-
nus only resupinate species with colored echinulate spores.
3. To restrict Stereum to pileate species which have simple
basidia and colorless spores and lack sctse in the
hymenium.
4. To include in Hymenochccte all species having setse.
5. To include in Coriicium species always resupinate, which
have colorless spores and lack cystidia, excepting those
species which for other reasons are placed in Exobasidium.
Include in Coriicium hypochnoid as well as compact
species.
6. To include in Peniophora all species which differ from
Coriicium merely by the presence of cystidia.
I find this system workable and very satisfactory for the
accurate location of species in genera, except in the case of the
species intermediate between Peniophora and Coriicium. The
proposals to subdivide Peniophora into Gloeocysiidium, Penio-
phorella, GlGeopeniophora, etc., would create large numbers of
species intermediate between the new genera, without compen-
sating advantages.
I have studied the species of my predecessors and co-workers
sympathetically and have endeavored to find real differences
between their species and those previously known so that the
validity of theirs might be confirmed. The great area of land
covered by the present work, the differences in climate and
substratum, and the keen search by my correspondents have
brought to hand a very large number of specimens. I have
earnestly striven to place them under species already known,
but it has been necessary to describe many as new. I regret
that there are so many of these. Should any one have reason
to believe that in any case I have described as new a species
already known, I shall esteem it a favor to receive an authentic
specimen of the older species or to be informed where such a
specimen can be consulted.
[Vol. 1
194 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Colors of specimens were noted and recorded during the first
years of my work by comparison with Saccardo's 'Chromo-
taxia' in accordance with his descriptive terms. Recently I
have been using Ridgway's 'Color Standards and Nomen-
clature/ 1912, which has a greater variety of colors useful in
the characterization of the species of Thelephoracece.
In my own work with collections of living fungi I am endeav-
oring to gather for each species a spore collection on a glass
slip. The spores adhere well so that they may be covered by
paper and preserved in the envelope mth the dried specimens
from which the spores were obtained. Such collections give
the exact color and dimensions of mature spores. These dimen-
sions are generally rather larger than those obtained from spores
of sectional preparations of dried herbarium specimens. The
spores of dried specimens, i. e., those remaining attached to the
specimens, are probably too immature to be of normal size, and
sometimes there arc so few of them that one must exercise
caution to avoid errors due to the study of spores foreign to the
fungus.
Latex exists in many species of several of the genera and is
more abundant and cons])icuous in some species than in others,
and its containing elements often extend to the hymenial surface.
When specimens are in the vegetative condition, injury to the
hymenium may liberate the fluid contents of the latex bodies
so that this fluid exudes in colored drops at the edges of the
wound, or discolors the bruised surface. For many of our species
there is a lack of data concerning the color of this fluid or
the discoloration. The latex bodies are pale brown in micro-
scopic preparations made by my methods and must not be con-
fused with setae or cystidia. Latex is well shown in Stereum
spadiceum, S. sanguinolentum , and Corticium lactescens.
There has been a disposition on the part of some authors to
regard the Thelephoracece as not sharply separated from the
Hyphomycetes. The specimens which I have collected, in striv-
ing to find all the Thelephoracem of my collecting region, and the
specimens received from my correspondents afford no embar-
rassment in recognizing the most hypochnoid species of Thele-
phoracece by the basidia which characterize the families of
Hymenomycetes in g(jneral.
1914]
BURT — TIIELEPHORACE.E OF NORTH AMERICA, I 195
The microscopical technique has been simpUfied as much as
possible. Usually dried herbarium material had to be used for
study and proved very satisfactory except in the case of speci-
mens which had been subjected to poisoning processes for pres-
ervation in herbaria. A small bit of the fructification having
a promising hymenial surface 2 or 3mm. square — but smaller if
the specimen is a valuable type — is first moistened with alcohol,
then wet with water and cut out from the rest of the specimen
and from the substratum. This bit is then placed in a holder
of elder pith and oriented so that the sections may be cut per-
pendicular to the surface of the hymenium and also contain as
long hyphse as possible. The sections are cut as thin as possible,
free hand, with a very keen section razor flooded with alcohol.
The thinnest sections are placed on a slide in a drop of water
and then a drop of seven per cent aqueous solution of potassium
hydrate is added.
Close observation of the sections should be made when the
potassium hydrate solution comes in contact with them. For
most species, the sections are merely cleared and the hyphse
swelled to the normal size of vegetative hyphse. In a few specieS;
the alkaline solution may dissolve out the color of the section
on coming in contact with it, or it may change this color to a
violet, which finally disappears, or it may cause disorganization
changes in certain structures leading to their disappearance or
destruction. Such changes should be observed and noted, for
they are of help in the determination of the species. In the
cases in w^hich potassium hydrate solution exerts a destructive
action, lactic acid should be employed with other sections in
the manner described for potassium hydrate. Lactic acid
clears and swells sections well, but so much more slowly than
potassium hydrate that I have used it only where the latter is
not satisfactory. After the sections have been cleared, the
potassium hydrate should be drained off, the sections lightly
stained on the slide with alcoholic solution of eosin (but not
overstained), mounted in water, and studied at once.
For a thorough study of the species of the family at least one
permanent preparation of each species should be retained for
future comparisons. Permanent preparations may be made
from the temporary water mounts by adding dilute glycerin
196 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
two-thirds glycerin and one-third water— at the edge of the
cover glass and allowing the glycerin to run under the latter as
the water evaporates. When concentration of the glycerin is
adequate, the excess should be wiped away with moist filter
paper and the resulting smear removed to the very edge of the
cover glass with a soft cloth moistened with 95 per cent alcohol.
The preparations may then be sealed from the atmosphere by
painting a ring of microscopical cement about the edge of the
cover glass. At least two coats should be used for this ring, a
hght and very narrow one, and, after this dries, a very heavy,
broad one. I have used Bell's Microscopical Cement, made in
London, and Brunswick Black Cement. A variable percentage
of the rings crack in the course of a few years and allow the
glycerin to escape from under the cover glass, but the sections in
such preparations can be remounted. Dr. Thaxter has very
recently informed me that he has been using King's Transparent
White Cement and King's Amber Cement for fifteen years and
that none of the rings made with these cements have cracked.
By the use of circular cover glasses rather than square ones,
a microscopist's turn table may be used, thereby materially
lessening the labor of preparing the rings.
Systematic Account
THELEPHORACEAE
Thelephorece Persoon, Myc. Eur. i: 109. 1822; Fries,
Hym. Eur. 629. 1872; Saccardo, Syll. Fung. 6:513. 1888.
Hymenomycetes with the hymenium inferior or amphigenous
(on the lower surface or surrounding the fructification), cori-
aceous or waxy, even, rarely ribbed or papillate.
Through several of the genera the TJielephoraceoB connect
closely with all tlie other families of the Hymenomycetes.
Hypochnus approaches Grandinia of the Hydnacece in the gran-
ular hymenial surface of many of the species, but can be sepa-
rated from this hydnaceous genus by the spore characters.
Lachnodadium , with coriaceous structure, hairy stem, and
colorless spores, is an intermediate genus between Clavaria, of
the Clavariacem, and ThelepJwra but can be separated from the
latter by the spore characters. Craterellus connects with
1914]
BURT — THELEPHORACE.E OF NORTH AMERICA. I 197
C anthar ellus , of the Agaricacece. Some species of Corticium must
be cautiously separated from Merulius, of the Polyporacece.
The species of Tremellodendron, Hirneolina, and Sehacma were
formerly distributed among Thelephora, Stereum, and Corti-
cium respectively, but are now separated from these genera by
the cruciate character of the basidia, — such basidia as are pres-
ent in many Tremellacece. All these connecting genera will
be included in the present monograph.
Michenera and Heterohasidium are excluded genera. Lyman
has shown^ that Michenera artocreas B. & C. is only a stage in
the life history of Corticiuin subgiganteum B. & C, and that the
genus Michenera has ceased to be a genus of the Basidiomycetes.
My own study of the type of Heterohasidium chlorascens Massee,
which is the type species of the genus, failed to locate any
basidia whatever.
Very many ThelephoracecB are of great economic importance
on account of the dry rot induced by the growth of the mycelium
in sills, floors, mine, bridge, and dock timbers, and other wooden
structures located in moist, poorly ventilated places. Conio-
phora puteana is a common species which rots coniferous wood.
Only a very few Thelephoracece are classed as serious plant
parasites. Of these the rhizoctonial stage of Corticium vagum
is the most important.
4
Key to the Genera
I. EU-THELEPHOREiE:
Fructification not containing green lichen gonidia.
Fructification fleshy or membranaceous, often infundibuliform, with the
hymenium distinct, continuous, even, ribbed or at length rugose; ba-
sidia simple Cralerellus
Fructification submembranaceous, cup-shaped, often pendulous; hymenium
typically concave, discoid; basidia simple Cyphella
Fructification consisting of only a fleshy hymenium on the surface of Uving
leaves and shoots; basidia simple Exohasidium
Fructification coriaceous or hard 1
1. Basidia globose or p^Tiform, longitudinally cruciately 4-septate or divided
when mature; fructification erect, clavariform, more or less branched .......
Tremellodendron
1 Cultural studies on the polymorphism of Hymenomycetea, Proc. Boston Soc. Nat. Hist. 33 1
151-60. 1907.
[Vol. 1
198 ANNALS OF THE MISSOURI BOTANICAL GARDEN
L Basidia cruciate as in Tremellodendron; fructification effuso-reflexed or cup-
shaped with the niar^in free Eirneolina
1. Basidia cruciate as in Tremellodendron; fructification ahvays resupinate.5e{>ac^^
1. Basidia simple but with such large sterigmata as to resemble lon^ntudinally
divided ba^idia^ Tulasnella
1. Basidia at first globose and simple, at length elongated and transversely v*?ep-
tate, straight or curved, bearing sterigmata on the convex side; fructification
resupinate Scptobasidium
1. Basidia simple, usually 4-spored 2
2. Spores colored; fructification pileate Thelepl
lora
2. Spores colored, rough-walled to echiuulate; fructification resupinate
Hypochnus
2. Spores ochraceous, ferruginous or fuscous, even; fructification resupinate . .
Coniophora
2. Spores white or rarely bright colored, even or rarely uneven 3
3. Seta) (bro^\Ta, cylindric, rigid, even-walled bodies) present in the hynienium;
fructifications range from pileate to resupinate Hyinenochcete
3. Cylindric teeth composed of many consolidated hypha^ protrude from the
hymenium but are not covered by it. Our southern species was originally
described as a Ihjdnum Mycdbonia
8. Neither setae nor teeth present in the hymenium 4
4. Fructification coriaceous, erect, clavariform; stem often hairy. .Lachnocladium
4. Fructification cup-sliaped, resupinate with free margin or simply resupi-
nate; hymenium pulverulent; with some two or three of the following
characters: (1) large white spores ranging from 14-34 x 12-20 m; (2)much
granular matter in the fructification; (3) prominent moniliform or
branched paraphyscs; (4) racemose organs in the hymenium which pro-
duce a crop of conidia before basidiosi)ores develop Aleurodiscus
4. Fructification pileate ranging from infundibuhform and flabelliform to
very narrowly reflex ed forms; hymenium even. Some reflexed species
may occur resupinate Slereum
4. Fructification like that of an urn-shaped ^S/crc urn but hard and stuffed.
One tropical species Hypohjssus
4. Fructification like that of Slereum but with the hymenium hardened and
with radiating branched ribs. Species tropical Cladoderris
4. Fructification always resupinate; structure not as in Alcurodiscus 6
5. Subhymenial tissue contains conspicuous brown stellate organs composed of
several radiating arms Asleroslroma
5. Such brown stellate organs not present 6
6. Cystidia present in hymenium, or in subhymenial tissue, or in both;
structure may be compact or hypochnoid Peniophora
6. Cystidia not present; structure compact or hypochnoid Corticium
t With regard to the nature of these bodies see 11. O. Juel. Bihang till K. Sv. Vet.-Akad. Handl.
23": Afd. III. 3-27. 1897.
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 199
II. HYMENO-LICHEXS:
Fructification regularly containing green lichen gonidia.
Species tropical.
Fructification pileate, coriaceous-membranaceous, with hymenium on the
lower surface and somewhat waxy; gonidial layer composed of some-
what cubical masses of algal cells ^'^^^
Fructification like Cora in most respects but with the hymenium somewhat
gelatinous and the gonidial layer composed of algal cells arranged in
(cateniform) Rhipidonema
rows
THELEPHORA
Thelephora EhrhsiTi [Crypt. Exsic. No. 178. 1785] Fries,
Syst. Myc. i: 428. 1821 (in part).-Persoon, Myc. Eur. i:
110. 1822 (in part).-Saccardo, Syll. Fung. 6: 521. 1888 (in
part) .-Hennings, in Engl. &. Prantl, Nat. Pflanzenfam. (i. 1**):
125. 1898 (in part).
The type species of the genus is Thelephora terrestris Ehrh.
ex Fries.
Fructifications pileate or clavate, coriaceous; hymenium con-
tinuous with the hymenophore and similar to it, inferior, or
amphigenous in a few species, even or faintly ribbed or papillose;
basidia simple, 4-spored; spores colored, typically muricate but
even, or rough-walled in a few species.
As more broadly defined by Fries and the other authors cited,
Thelephora has been heterogeneous, consisting chiefly of the
natural and homogeneous group of species defined above but
also of some pileate species with simple basidia and hyaline
spores, transferred to Stereum; also of some species with globose,
longitudinally septate basidia, transferred to Tremellodendron,
if with erect fructifications, or to Sehacina, if resupinate; and
also of some resupinate species having simple basidia, of which
those with muricate and colored spores may be found in Hypoch-
nus, those with colored and even spores, in Coniophora, and
those with hyaUne spores, in Corticium and Peniophora.
probable that the species of Fatouillard's section Dendrocladium
of the genus Lachnocladium as understood by Patouillard ^
jht be transferred to Thelephora with advantage both to
Thelephora and Lachnocladium, but these species are not within
the geographical limits of my work.
» Fragments Mycologiques (suite). Jour, de Bot. 3:33-37. 1889.
It
mi
[Vol. 1
200 ANNALS OF THE MISSOUBI BOTANICAL GARDEN
Key to the Species
Erect species, usually with central stem and pileus divided into very narrow,
brandling, flattened or cylindric divisions; hymcnium inferior or am-
phigenous
Erect species, usually with central stem and more or less infundibuliform,
cup-shaped or flabellifonn pileus, wiiich may be radially split into
1
lobes and divisions 2
Species of incrusting, efTuso-reflexed, dimidiate, or applanate habit 5
1. 2-6 cm. high, much branched, glabrous, with fetid odor when growing,
perhaps rarely odorless t.T. yalmata
1. 3-5 cm. high, much branched, minutely pubescent; stem villose, without fetid
odor. Compare T, mulivpariiia 2, T, aiithoccphala
1. Less than 2| cm. high, branching at or below surface of ground, dusky drab
except at base s. T. ojespiiulans
1. Less than 2 cm. high, very slender and fragile, cinereous. Known only from
State of Washington 4. 7\ scissilis
1. Large species, highly branched, with body of spore of regular obovoid form.
Known only from Central America 5. T, angiisiata
2. Hymcnium dark colored, i. e., brown to fuscous 3
2. Hymcnium light colored, i. e., pallid to gray 4
3. Small species, l|-3 cm. high, upper surface usually drying pallid, usually
deeply cleft or many-parted into narrow divisions; stem villosc^. T . muUrpartita
3. Small species, 6 mm.-2J cm. high, infundibuliform or deeply divided into
two or three triangular divisions, or flabelliform; stem villose. Closely related
to T, muUiparlita 7. y. regularis
3. Fructification 1 cm. high, white; stem white, glabrous. Known only from
Guadaloupe s, T. pusiola
3. 1^—5 cm. high, larger species than the three preceding but with thinner pileus,
fuscous purple (Rood's brown) throughout, often with the thin lobes imbri-
cate like the petals of a carnation; stem villose 9, T. caryophylka
3. 2-4 cm. high, somewhat tubular, hymcnium vinaceous brown to drab; stem
fiulcate and pitted but not villose; spores 10-14 fx in diameter. Known only
from Jamaica lo. T. magnispora
3. Large species, 2J-7 cm. in diameter, with upper surface pallid except at the
center and with the hymeniimi dark IS.T. malts
4. Small species, less than 2 cm. in height and in diameter, somewhat pallid
to brick-red 7. t. regularis
4. Pileug with outer lobes forming a cup and with inner lobes distinct,
crowded, erect, cinereo-fuscoue. Known from Costa Rica and Brazil.
11, T. corbifonnis
4. Large species, 5-7 cm. broad, deeply infundibuliform, habit and color of
Crater cllusconiiicopioidcs. Costa Rica and Jamaica. ./;?. T. cornucopimdes
5. Growing in applanate clusters, effuso-reflcxed, or dimidiate 6
5. Always incrusting {T, albido-brunnea is sometimes incrusting) g
6. Hymenium pale and colored like the pileus, cinnamon-buff; pileus
6-8
and
15, r. lutosa
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. T 201
6. Hymenium drab, becoming sage-green when crushed in 7 per cent potas-
sium hydrate solution; pileus pinkish buff to omnamon-brown with a
broad pale margin 16. T. cuticularis
6. HjTnenium ferruginous brown (Rood's brown) to fuscous ' 7
7. Pileus, when squamulose, with the fibers matted and agglutinated into ap-
pressed and wholly adnate squamules, margin dilated and whitish fimbriate
becoming entire and concolorous 17. T, intyhacea
7.^Pileu3 not zonate, fibrous-squamulose and usually strigose, margin fibrous-
fimbriate 18, T, terrestris
7, Pileus zonate, in other respects resembling the preceding species
19. T, griseozonata
8. Incrusting and ascending small plants, free branches somewhat terete but
flattened towards the tips; spores umbrinous SO. T.fimbriata
8. Resupinate on leaves and twigson the ground and sending up free, simple
or branching trunks; spores fuscous. Known from Cuba only 21 , T, perplexa
8. Incrusting leaves, etc., on the ground and ascending as sessile flabclhform
pilei which are dentate at the upper end or deeply divided, honey-
yellow to tawny olivaceous throughout. Known from Cuba only. . . .
SS. T. denlosa
8. Typically effused, rising obliquely upward from the support as a cluster of
small trunks which branch and terminate in spiculous tips. ^5. T. spiculosa
!• Thelephora palmata Scop, ex Fries, Syst. Myc- i:432.
1821.
Plate 4. fig. 4.
Clavaria palmata Scop. FL Carn. 2: 483, 17(jO~Ramaria
palmata Holmsk. Fun. Dan. i: 106. pL — .1799. -Merisma
foetidum Pers. Syn. Fung. 584. 1801. -M. palmata Pcrs. Myc.
Eur. i;113. 1S22 -Thelephora palmata americana Peck, Rep.
N. Y. State Mus. 53: 857. 1900.
Illustrations: Greville, Crypt. FL i : pi. ^^.-Holmskiold,
Fun. Dan. i : pi. of Ramaria 'pahnata.-KTombholz, Abbild. und
Beschr. jpl. 5Jf.f.2Jf, ^5.-Nees, System 2??- 16. f. 151 ^.-Baillon,
Dictionn. de Botan. i : 737. /. 7.-Loudon, Encyc. of Plants
/. 16131.-WmiQv, Crypt. Flora i: 321.
Fructification coriaceous-soft, fuscous purple, drying cinna-
bar-brown or chestnut-brown, erect, very much branched, with
very fetid odor; pileus with numerous somewhat fastigiate,
palmate divisions which are even, flattened, dilated above, and
with fimbriate and whitish tips; stem simple or soon branched;
hymenium amphigenous; spores pale umbrinous under the
microscope, sparingly echinulate,10 x 7-8 n.
Fructification of American specimens 2-6 cm. high, 1-3 cm.
broad; stem 1-1| cm. long, 1-2 mm. thick.
[Vol. 1
202 ANNALS OF THE AIISSOURI BOTANICAL GARDEN
On moist ground in coniferous woods and also in grassy fields.
Prince Edward Island to North Carolina and west to Illinois.
June to October.
In the American collections of this species the divisions of the
pileus are narrow and a short stem is present. The habit is so
similar to that of Thelephora anthocephala that record of the
fetid odor should always be made if observed. The ultimate
branches may be more or less terete, leading to the variety
amcricana Pk.
Specimens examined:
Exsiccati: Ell. & Ev., N. Am. Fungi, 1937.
Austria: G. Bresadola.^
Sweden: L. Romell, 53.
Canada: Rustico Bay, Prince Edward Island, J. Macouri, 3^4.
New Hampshire: Chocorua, W. G. Farlow.
? Vermont: no locality data for specimen in Frost Herb., Univ.
of Vermont.
Connecticut: Manchester, C. C. Hanmer, 1398.
New York: Fischer's Island, C. C. Hanmer, 196.
New Jersey: C. G. Lloyd, 4612.
Pennsylvania: Bethlehem, Schweinitz, Syn. N. Am. Fungi, 612
(in Herb. Schw.) ; Trexlertown, Dr. W. Herbst; Kitanning,
D. R. Sumstine, 2; West Chester, B. M. Evcrhart, Ell. &
Ev., N. Am. Fungi, 1937.
Delaware: Newark, H. S. Jackson.
Dist. of Columbia: Washington, 0. F. Cook, comm. by P. L.
Ricker, 1, 3.
N. Carolina: Asheville, H. C. Beardslee, 924-
Ohio : Connecticut, C. G. Lloyd, 4493.
Illinois: Glencoe, E. T. and S. A. Harper, 664, 665.
Missouri: St. Louis, N. M. Glatfelter (in Mo. Bot. Card. Herb.,
42560).
1 With regard to the citation of specimens all except those of "Exsiccati" are
in Burt Herb, which are cited without explicit reference to place in other herbaria.
For example, the specimen cited, "Connecticut: Manchester, C. C. Hanmer, 196,"
is in Burt Herb. The data given is that received with the specimen and may identify
a duplicate in another herbarium. The location of all specimens in herbaria other
than my own is designated by the name of the herbarium in parenthesis with the
prefix "in." For example, the specimen cited, "Louisiana: St. Martinville, A. B.
Langlois (in Lloyd Herb., 3000)," is in Lloyd Herb., but not in Burt Herb.
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 203
2. T. anthocephala Bull, ex Fries, Syst. Myc. i: 433. 1821.
Plate 4. fig. 1.
Clavaria anthocephala Bull. Herb, de la France 2 : 197, pi. 1^52.
/. 1. 1789.
Illustrations: Bulliard, Ihid. pi. 4^ 2 . f . 1 .-^ow eihy , Col.
Figs. Eng. Fun. pi. ^^^.-Berkeley, Outlines Brit. Fung. pi.
17. f. ^.-Dufour, Atlas des Champ, pi. 70.
Fructification coriaceous-soft, somewhat ferruginous, drying
fawn-color or cinnamon-brown, inodorous; pileus pubescent,
divided to the stem into flaps which are dilated upwards and
fimbriate and whitish at the apex or divided into irregular,
branched, erect branches; stem simple, equal, villose; hymenium
even; spores pale umbrinous under the microscope, ranging from
angular-tuberculate to tuberculate-echinulate, 8-10 x 7-8iu.
Fructifications 3-5 cm. high, 1-3 cm. broad; stem 1-1| cm.
long, 1-2 mm. thick.
On the ground in woods. Massachusetts and Ohio to Louisi-
ana. June to August. Rare.
Our specimens of T. anthocephala and T. palmata have the
same habit but may be separated, even when dried, by the fine
pubescence of the pileus visible with a lens, and by the villose-
tomentose stem of the former. The spores of T. anthocephala
are further slightly paler and have shorter spines with broader
bases than those of T. palmata.
Specimens examined:
Austria: G. Bresadola.
Massachusetts: Newton, W. G. Farlow (in Farlow Herb.).
New York: Van Cortlandt Park, N. Y. City, L. 0. Overholts
(in Overholts Herb., 688).
Pennsylvania: Kitanning, D. R. Sumstine, 10; Bethlehem,
Schiveinitz (in Herb. Schw.), the 614 of Syn. N. A. Fungi
under the name T. jldbellaris.
North Carolina: Asheville, H. C. Beardslee, 0268.
Louisiana: St. Martinville, A.B. Langlois, unnumbered specimen,
and 1971, and by the same collector (in Lloyd Herb., 3000).
Ohio: Norwood and Linwood, C. G. Lloyd, 1472 and 02164 re-
spectively.
Kentucky: C. G. Lloyd, 1395.
204 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[YOL. 1
Missouri: St. Louis, N. M. Glatfelter (in Mo. Bot. Gard. Herb.,
42559) .
3. T. caespitulans Schw. Trans. Am. Phil. Soc. N. S. 4: 166.
1831.1
Herb. Schw
Fructification erect, coriaceous, dusky drab to oUve-brown
below, paler above, very much branched, forming clusters 2|
cm. high by 2| cm. broad; pileus with numerous divisions joined
together into a solid base but assurgent above and pressed to-
gether closely, compressed, subcanaliculate, frequently obtuse
and whitish at the apex; hymenium amphigenous; spores um-
brinous under the microscope, sparingly tuberculate, 7-8 x
5-6/i.
On the ground in mixed woods, Vermont to South Carolina,
and in dense coniferous woods, Washington. September. Rare.
This species is related to T. palmata but is more olivaceous,
and it is probably inodorous, — at least no odor has been noted.
The dimensions for the clusters given above, as stated by
Schweinitz, are probably maximum dimensions, for the speci-
mens recently collected have been rather smaller. My Vermont
specimens were growing with the thick, solid base buried in
sandy ground in a wood road; they have dried palhd except at
the base and are slightly pubescent. The general habit of
this species is somewhat suggested by a small cluster of Tremel-
lodendron pallidum (Schw.) Atk.
Specimens examined:
Vermont: Lake Dunmore, E. A. Burt.
Pennsylvania: Bethlehem, Schweinitz, type (in Herb. Schw.,
Acad. Nat. Sci., Phila.).
South Carolina: Santee Canal, Ravenel, 1660 (in Curtis Herb.
under name T. vialis).
Washington: Chebalis, C. /. Humphrey, 1287; Bingen, W. N.
Suksdorf, 689.
4. T. scissilis Burt, n. sp. Plate 4. fig. 8.
Type: in Burt Herb.
Fructifications gregarious, coriaceous, erect, clavariform,
branched, longitudinally ridged by the bases of numerous,
' A figure will be given in Part II.
1914]
BURT— THELEPHORACE^ OF NORTH AMERICA, I 205
small, appressed, acicular branches, the larger of which are at
the apex of the fructification and spread slightly outward in
fan-shaped manner; stem glabrous, castaneous; hymenium am-
phigenous, on upper half of the fructification, avellaneo-ciner-
eous; basidia simple, hyaline, 4-spored; spores pale umbrinous
under the microscope, angular, 6-8 x 5-6/i.
Fructifications l§-2 cm. high; spread of branches at the top
2-6 mm.; stem 7-10 mm. long, 1 mm. thick.
On the ground. Washington. January.
This species is very distinct by its slender erect habit, cinereous
color, and only slightly spreading branches.
Specimens examined:
Washington: Bingen, KHckitat Co., W. N. Suhsdorf, 716, type.
5. T. angustata Fries, (Nov. Symb. Myc. 92.) Actis R. Soc.
Sc. Upsal. III. i: 108. 1851.
Type: in Herb. Fries.
f
Fructification erect, cinereo-fuscous, pliant, becoming rigid
and somewhat woody; stem elongated, radicated, rugose, gla-
brous, compressed, irregularly divided at the upper end into
unequal, fastigiate, compressed branches, which are clothed on
the whole outer surface with the hymenium; hymenium amphi-
genous, subrugose, gray; basidia simple; spores umbrinous under
the microscope, obovoid, apiculate at base, flattened on one
side, echinulate, 14 x 7-9At.
On decaying wood. Central America.
Substance, color, and hymenium exactly as in T. cornuco-
pioides, but of the very different form of Clavaria rugosa and
having highly branched forms; stem 5 cm. long; color fuliginous.
The fructification is fleshy-pliant when fresh, but on drying
hardens much more than species of Stereum.
Specimens examined:
Costa Rica: Oersted (in Herb. Fries), type.
6. T. multipartita Schw. in Fries, Elenchus Fung, i : 166.
1828.
Type: in Herb. Schweinitz.
Plate 4. fig. 7a.
Fructifications gregarious, erect, coriaceous, fusco-cinereous,
usually drying pallid; pileus infundibuliform, sometimes cleft
4
[Vol. 1
206 ANNALS OF THE MISSOURI BOTANICAL GARDEN
more or less deeply and unequally into a few lobes, sometimes
divided to the stem and spreading so as to appear dimidiate,
very often deeply divided and subdivided into many narrow
and spreading divisions more or less dilated and whitish at the
apex; stem erect or incurved, equal or tapering upward, some-
times branched above, drying walnut-brown or palUd, villose;
m
hymenium inferior, glabrous, even, fawn-color or vinaceous
drab; spores unbrinous under the microscope, tuberculate, 7-9
X 5-6/1.
*
Fructification lf-3| cm. high, 1-3 cm. broad; stem 1-2 cm.
long, 1-3 mm. thick.
On ground in groves of broad-leaved trees, especially under
oak. New York and Pennsylvania to Illinois. July to Sep-
tember.
The upper surface of the pileus was originally described as
glabrous, but it is minutely pubescent under a lens, or sometimes
fibrillose. This species is very perplexing by its close relation-
ship to T. regularis. The multipartite pileus is the only char-
acter which seems available to separate collections of the former
from the latter species. If a given collection consists wholly
of specimens with pileus many-parted and subdivided into nar-
row divisions, or if it contains some such specimens in addition
to others with more regular infundibuliform pileus, I refer the
collection to T. multipartita, as in the cases of the collections
cited below from C. 0. Smith and Dr. C. H. Peck respectively.
As yet, I know of no characters by which to assort and separate
into their respective species specimens mixed together of typical
T. regularis and those specimens of T. multipartita which have
the pileus infundibuliform or merely cleft more or less deeply
and unequally into a few lobes* Therefore it is my opinion
that T. multipartita is a variety of T. regularis^ but the collec-
tions which have so far been submitted to me, have been com-
posed of too few fructifications to assure me that this opinion
is correct.
Specimens examined:
Exsiccati: Ell. & Ev., N. Am. Fungi, 280G, under the name T.
caryophyllea.
1914]
BURT — THELEPHORACE.I; OF NORTH AMERICA. I 207
New York: Bolton, C. H. Peck, S, 4, 5; Ithaca, C. 0. Smith,
Cornell Univ. Herb., 13359, and C. 0. Smith and W. H.
Long, Cornell Univ. Herb., 7743.
New Jersey: Newfield, /. B. Ellis, Ell. & Ev., N. Am. Fungi,
2806.
Pennsylvania: on island in Lehigh River, Schweinitz, type (in
Herb. Schw.); Bethlehem, Schweinitz (in Herb. Schw.),
the T. tuber osa of Syn. N. Am. Fungi, 613; Trexlertown,
W. Herhst, n, 36.
Ohio: A. P. Morgan, Lloyd Herb., 2581, 2647; Oxford, L. 0,
Overholts (in Overholts Herb., 1685).
Illinois: River Forest, E, T. and S. A. Harper, 666.
7. T. regularis Schw. Schrift. d. Naturforsch. Gesell., Leipzig,
Plate 4. figs. 6, 7b.
1 : 105. 1822.
Thelephora Ravenelii Berk. Grevillea i : 148. 1873.- r. hiscens
Berk. & Rav. Grevillea i : 148. 1873.
Type: in Herb. Schweinitz, Acad. Nat. Sci., Phila.
Pileus coriaceous, solitary, infundibuHform or divided to the
stem into triangular divisions or flabeUiform, fibrillose, drying
pallid or tawny-olive, darker at center of the cup or at base of
the divisions, margin lacerate; hymenium usually hair-brown,
sometimes pallid; spores melleus to umbrinous under the micro-
scope, angular-tub erculate, 6-7 x 4^-5/x.
Fructification 6 mm.-2| cm. high; pileus 5 mm.-2| cm. broad;
stem 3-15 mm. long, 1-1^ mm. thick.
In moss in wet places and on humus. Ontario to Alabama
and westward to Kansas.
The differences in form of the pileus of T. regularis are well
shown by the type in Herb. Schweinitz; this type consists of
three fructifications, two of which are infundibuHform, the
third and largest, flabeUiform. The hymenium is sometimes
merely pallid, as in the case of the specimen which is the T,
pannosa of Schweinitz, Syn. N. Am. Fungi, No. 606, but is not
T. pannosa Fr. The cotypes of T. Ravenelii and T. hiscens
agree in all respects with the authentic specimen of T. regularis
in Curtis Herb. Specimens of T. regularis which have the pileus
infundibuHform and little cleft are suggestive of small specimens
of T. caryophyllea but differ from the latter by the thicker pileus
[Vol. 1
208 ANNALS OF THE MISSOURI BOTANICAL GARDEN
and paler coloration of T. multipartita which is wholly lacking
in the rufescent coloration of T. caryophyllea. There is a diffe-
rence of form between specimens of these two species which is
brought out well by the figures in pi. 4.
Specimens examined:
Canada: Shannonville, Ontario, J, Macoun, 330.
Maine: Portage, L. W. Riddle, 4-
New Hampshire: Chocorua, W. G. Farlow (in Farlow Herb.).
Massachusetts: near Boston, Sprague, 246 (in Curtis Herb.
under the name T. anthocephala) ; Newton, W. G. Farlow
(in Farlow Herb.).
Pennsylvania: Bethlehem, Schweinitz, station cited by Schwei-
nitz; also the specimen (in Herb. Schw.) under the name
T. pannosa of Syn. N. Am. Fungi, No. 606; Trexlertown,
C. G. Lloyd; Kitanning, D. R. Sumstine.
Delaware: Clayton, H. S. Jackson.
North Carolina: Salem, Schweinitz, type (in Herb. Schw.);
G. F. Atkinson, Cornell Univ. Herb., 23254.
South Carolina: Greenville, Ravenel, 1498, type and cotype (in
Kew Herb, and in Curtis Herb, respectively) of T. Rav-
enelii Berk.; Santee Canal, Ravenel, type and cotype (in
Kew Herb, and in Curtis Herb, respectively) of T. hiscens
Berk. & Rav.
Alabama: Peters, 576 his (in Curtis Herb, under the name
T. anthocephala).
Wisconsin: Madison, W. Trelease (in Farlow Herb.); Lake
Geneva, E. T. and S. A. Harper, 882, and (in Harper Herb.,
883).
Illinois: East St. Louis, A''. M. Glatfelter (in Mo. Bot. Gard.
Herb., 42563).
Iowa: Johnson County, T. J. Fitzpatrick, 39.
Missouri: St. Louis, N. M. Glatfelter (in Mo. Bot. Gard. Herb.,
42564).
Kansas: Bourbon County, A. 0. Garrett, 80.
8. T. pusiola Pat. in Duss, Champ. Guad. & Martinique 12.
1903.
Pileus with divisions triangular, white, hard, thin, entire or
cut4obed, glabrous, even or rugose, sometimes zonate, atten-
1914]
BUET — THELEPHORACEjE OF NORTH AMERICA. I 209
uated into a slender stem; stem colored like the pileus, glabrous,
cylindric, woody; hymenium inferior; even, brown; basidia clav-
ate, 25 x lO/i, four-spored; spores globose-angular, colorless or
somewhat fuliginous, 6/x in diameter; no cystidia.
Fructification 1 cm. high, divisions 5 mm. broad.
Solitary or in clusters on dead wood. Guadaloupe. Forest
of Bains-Jaune, Duss, 589.
Var. terrestris Pat. Ibid, has the divisions of the pileus nar-
rower, laciniate, divergent, rigid.
On the ground, Matouba, Guadaloupe, Duss.
I have seen no specimens of either this species or its variety,
neither of which have been reported since their original dis-
cover3^
9. T. caryophyllea Schaeffer ex Fries, Syst. Myc, i : 430.
1821. Plate 4. fig. 9.
Elvella caryophyllea Schaeffer, Icon. Fung, 3: ll5.pl. 325.
17 62-17 7 4.~Craterella ambigua Pers. Obs. Myc. i: 36. pi. 6.
/. 8-10. 17 9Q.-Thelephora caryophyllea 7 amhigua Pers. Myc.
Eur. i: 112. 1822.
Illustrations: Schaeffer, Icon. Fung. pi. 5^5.-Persoon, Obs.
Myc. i: pi. 6.f. (5-iO.-Schnizlein, in Sturm, Deutsch. Flora 3:
m
fasc. 31. pi. ^.-Lanzi, Fungi di Roma pL 11. f. -^-Saunders
and Smith, Mfe^c. 111. pi. 41.f. 7-12.-^mith, W. G. Brit. Basid.
399./. 96 a, h.
Fructifications solitary or cespitose, coriaceous, fuscous purple
but drying wood-brown; pileus infundibuliform, simple, or
doubled by proliferous growth of smaller pilei from the disk of
the principal pileus or of wedge-shaped lobes rising from its
upper surface, upper surface radiately ridged or striate with
masses of agglutinated fibers which are often dark colored, ob-
scurely zonate when moist, margin incised ; stem usually central,
cylindric, villose, simple or branched; hymenium inferior, even,
grayish olive to light yellowish olive; spores pale umbrinous,
tuberculate, 7-8 x 6/*.
Fructification l|-5 cm. high, 1^-5 cm. broad; stem 1 cm. long,
2-3 mm. thick.
On the ground under pines. Canada to South Carolina and
west to Ohio, also in the Pacific states. August to November.
Abundant locally.
210 ANNALS or THE MISSOURI BOTANICAL GARDEN
[Vol. 1
T. caryophyllea may be distinguished from our other northern
species which have a central stem and dark hymenium, by the
thin lobes of the pileus which dry paler than the hymenium,
and by the frequent occurrence of specimens with the pileus
consisting of many lobes and pilei imbricately arranged in a
manner suggestive of a double pink or carnation, as shown by
Schaeffer's fig. 5, and Persoon's fig. 10 of the illustrations cited.
Our specimens agree well with the figures of SchaefTer and Per-
soon — those of Persoon are especially good but unfortunately
occur in a work which is very rare.
We find occasionally specimens which agree well with T.
radiata (Holmsk.) Fr., but these specimens are connected so
closely by intermediate forms — often in the same collection —
with others which are undoubtedly T. caryophyllea that I refer
them to the latter species.
Specimens examined:
Sweden: K. Starhach, in Romell, Fun. Scand., 121.
Canada: /. Macoun, 54 and 75 of 1903.
Quebec: Hull, /. Macoun, 190.
Ontario: London, J. Dearness (in Lloyd Herb.).
New Brunswick: Restigouche River, T. F. Allen, comm. by
Dr. Farlow.
Maine: Orono, L. W. Riddle, 9.
New Hampshire: Shelburne, W. G. Farlow.
Vermont: Newfane, C. D. Howe; Middlebury, E. A. Burt, four
collections.
Massachusetts: Sprague, 47, Russell, 131, and D. Murray, 545
(all in Curtis Herb.); Worcester, G. E. Francis, 105.
Connecticut: East Hartford, C. C. Hanmer, i44^; Central Vil-
lage, J. L. Sheldon, 68, comm. by New York Bot. Card.
New York: Bolton, C. H. Peck; Ithaca, G, F. Atkinson, 9993,
9994; Saranac Lake, E. A. Burt; East Galway, E. A. Burt.
Pennsylvania: Bethlehem, Schweinitz (in Herb. Schw.), the 608
of Syn. N. Am. Fungi.
Dist. of Columbia: Zoological Park, Coville and Cook, No. A,
comm. by P. L. Ricker.
North Carolina: Schweinitz (in Herb. Schw.).
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 211
Michigan: C. G. Lloyd, 4547; Sailor's Encampment, E. T. and
S. A. Harper, 439, and Univ. of Wis. Herb., 2.
Ohio: C. G. Lloyd, 1422, 2720; Cincinnati, A. P. Morgan, Lloyd
Herb., 2641, and (in Lloyd Herb., 1152); Loveland, D. L.
James (in Herb. U. S. Dept. Ag.).
Kentucky: C. G. Lloyd, 1152.
Washington: Bingen, W. N. Suksdorf, 717, 690.
California: Jackson, J. H. Barber, comm. by W. A. Setchell;
Stanford University, C. F. Baker, 255, distributed by Baker,
Pacific Slope Fungi, 3743, under the name T. radiata
(Holmsk.) Fr.
■
10. T. magnispora Burt, n. sp. Plate 4. fig. 5.
Type: in Burt Herb.
Fructifications solitary or gregarious, coriaceous, stipitate;
pileus irregularly infundibuliform, somewhat tubular, with
ascending recurved lobes, drying avellaneous, becoming fuscous
at the center with age, fibrous torn becoming radiately striate,
margin incised; stem equal, soUd, drying hard, irregularly angled,
sulcate and pitted, vinaceous brown to drab; hymenium inferior,
even, vinaceous brown; basidia simple; spores pale cinnamon,
subglobose, echinulate, 10-14 /x in diameter.
Fructification 2-4 cm. high; pileus 1-2 cm. in diameter; stem
7-12 mm. long, 2-5 mm. thick.
On mossy ground. Chester Vale, Jamaica. December.
In some of the specimens the pileus is decidedly eccentric
through greater growth on one side than on the other, and it is
not always lobed. The offensive odor of the dried specimens
and the color of the hymenium are suggestive of T. cuticularis.
Specimens examined:
Jamaica: Chester Vale, W. A. and Edna L. Murrill, type, New
York Bot. Gard., Fungi of Jamaica, 295.
11. T. corbiformis Fries, (Nov. Symb. Myc. 92.) Actis. R.
Soc. Sc. Upsal. III. i: 108. 1851.-Romell, Hymenomycetes
Austro- American!. Bihang till K. Sv. Vet.-Akad. Handl. 26":
Afd. III. 44. 1901.
Type: in Herb. Fries.
Fructification sessile, rigid, cinereo-fuscous, with cespitose
lobes of which the outer ascend and coalesce into a rounded
212 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
cupulate pileus here and there lacunose-pervious, and the inner
are distinct, crowded, erect, narrow; hymenium inferior, uneven,
whitish ; basidia simple ; spores slightly colored, becoming uneven,
ovoid, 5-6 X 4-5 m.
On the ground. Costa Rica and Brazil. January.
''In substance, texture, color, etc., this species agrees exactly
with Thel. cornucopioides and Thel. angustata but in form it ex-
hibits a type unique in the Hymenomycetes. The clusters are
regularly rounded, very dense, divided all the way to the base
into innumerable lobes, of which the interior are free and erect,
the exterior regularly ascendant, broader, compressed, clothed
underneath by the hymenium and grown together into a cup
here and there lacunose-pervious, undulate-crisped at the apex
and fimbriate." — Translation of the original comment on this
species.
In 1899, I found the type in Herb. Fries to be cinereo-palhd
with a sHght fuscous tinge and with basidia and spores as stated
above but many of the spores even. Romell describes the spores
of his specimens from Brazil as "hyalinae, laeves, elHps., 5-7
X 3-4 mmm.," and as agreeing with the type. I have reexam-
ined my sections frora the type; the spores are certainly colored
and many of them rough-walled.
Specimens examined :
Costa Rica: San Jos^, Oersted (in Herb. Fries, Univ. Upsal.),
type.
12. T. cornucopioides Fries, (Nov. Symb. Myc. 91.) Actis
R. Soc. Sc. Upsal. III. i : 107. 1851. ^
Type: not known to be in existence; not in Herb. Fries, at
Upsala, nor in Kew Herb.
Pileus pliant becoming rigid, deeply infundibuliform, 5-7^
cm. broad, radiately rugose, glabrous, fuscous; stem solid,
rather glabrous, pallid; hymenium inferior, somewhat rugose,
gray.
On the ground. Near San Jos^, Costa Rica.
This species bears so singular a resemblance to Craterellus
cornucopioides that from pictures they are scarcely to be dis-
tinguished. The present species has the stem truly solid and
the substance fleshy pliant when living, nearly stony-woody
when dry; stem 5-7^ cm. long, 4-6 mm. thick, equal or attenu-
m
1 A figure will be given in Part II.
1914]
BURT — THELEPHORACEiE OF NORTH AMERICA. I 213
ated at the base, compressed, rather glabrous, very tough, pallid;
pileus membranaceous-cartilaginous, when dry quite rigid,
radiately rugose, with the ridges elevated towards the undulate
and at first fimbriate margin, not zonate after the manner of
species of Stereum; hymenium inferior, hardened. Related to
Cladoderris.
I refer to T. cornucopioides a collection made in Jamaica by
Prof. F. S. Earle, in 1902, the specimens of which agree well
with the original description, as translated above, except in size.
They are 3-3| cm. high and 2 cm. broad with stem about 1 cm.
long by 2-4 mm. thick. The dried fructification is very hard
and stony and softens so little with water that the edge of the
razor is turned in sectioning. The spores are colorless and even
at first and become slightly colored and angular, 9-10 x 6ai.
Specimens examined:
Jamaica: Castleton Gardens, F. S. Earle, New York Bot. Gard.,
Plants of Jamaica, 238.
13. T. vialis Schw. (Syn. N. Am. Fungi) Trans. Am. Phil.
Soc. N. S. 4: 165. 1834. Plate 5. fig. 15.
T. tephroleuca B. & C. Grevillea i :149. 1873.
Type: in Herb. Schweinitz.
Fructification coriaceous, dirty whitish or pallid, sometimes
wood-brown at the center, upper surface usually radiately pli-
cate or rough with masses of agglutinated fibers ; pileus polymor-
phic, sometimes composed of ascending lobes or small pilei which
arise from a common base and grow together above to form a
broad cup^ or sometimes with the whole interior of the cup filled
with small pilei and lobes many of which arise proliferously
from the upper surface of the outer lobes; stem central when
present; hymenium inferior, rugose, somewhat papillose, yel-
lowish pallid becoming avellaneous or somewhat fuscous;
spores olive-buff under the microscope, bluntly angular (i. e.,
tips of the angles obtuse), 4^-7 x 4|-5iu.
Fructification 2§-5 or 6 cm. high, 2|-7 cm. broad.
On ground in frondose woods. Vermont to South Carolina
and west to Illinois. September.
This is a fine, large species well marked by the dirty whitish or
yellowish, fibrillose upper surface of the pileus, thick substance
of the same color unless the specimen is old, and the brown,
[Vol. 1
214 ANNALS OF THE MISSOURI BOTANICAL GARDEN
slightly wrinkled hymenium. As in the otherwise very different
T. caryophyllea, large specimens sometimes resemble a double
flower from the great number of small pileoli and lobes present
in the center. Schweinitz described the species as sometimes
having dimidiate pilei, but I have seen no such specimens. My
collection assumed a disagreeable odor in drying but no such
odor has been noted by others.
Specimens examined:
Exsiccati: Ell. & Ev., N. Am. Fungi, 1110, and Fun. Col., 1593,
in both under the name T. caespitulans .
Vermont: Lake Dunmore, E. A. Burt.
New Jersey: Newfield, J. B. Ellis (in Mo. Bot. Gard. Herb.,
5155), also in the exsiccati cited.
Pennsylvania: Bethlehem, Schweinitz, type (in Herb. Schw.);
Michener, 1504 (in Curtis Herb, and in Kew Herb.), the
cotype and type respectively of T. tephroleuca; Trexlertown,
W. Herhst, 43, C, G. Lloyd and W. Herbst, 2866, 3088
(both in Lloyd Herb.) ; N. M. Glatfelter (in Mo. Bot. Gard.
Herb., 42561).
Dist. of Columbia: Washington, F. J. Braendle, comm. by C. H.
Peck.
North Carolina: G. F. Atkinson (in Cornell Univ. Herb., 23253);
Asheville, H. C. Beardslee; Schweinitz cited North Caro-
F '
lina as a station.
South Carolina: Caesar's Head, Ravenel, one of the types (in
Curtis Herb, and Kew Herb.) of T. tephroleuca.
Ohio : C. G. Lloyd, 4000.
Illinois: Glen Ellen, E. T. and S. A. Harper, 669.
14. T. albido-brunnea Schw. Trans. Am. Phil. Soc. N. S.
4:166. 1834.
Plate 5. fig. 13.
Stereum Micheneri B. & C. Grevillea i: 162. 1873 (in part).-
Stereufn spongiosum Massee, Jour. Linn. Soc. Bot. 27: 172.
lSS9.-Thelephora odorifera Peck, Rep. N. Y. State Mus. 44:
132 (22). 1891.
Type: in Herb. Schweinitz.
Pileus sessile or with very short stem, coriaceous, spongy when
dry, uniformly cinnamon-buff or with the older portions chest-
nut-brown, sometimes assuming mesopod form when encircling
small twigs or shrubs, sometimes effuso-reflexed, usually dimidi-
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 215
ate and somewhat imbricated, fibrous-tomentose, margin thick
and entire; substance concolorous with the upper surface,
spongy, more than 2 mm. thick, with hyphae 4^-5^^ in diam-
eter; hymenium inferior, even, not polished, cinnamon-buff;
basidia simple; spores deep olive-buff under the microscope,
echinulate, 8-10 x 6-8iU.
Pileus 2-4 cm. in diameter when circular, or 1-2| cm. long,
2-4 cm. broad, often 1 cm. thick at base when dimidiate.
Running up and encircling twigs on the ground and against
the base of shrubs. Canada to Louisiana and west to Wiscon-
sin. August.
Peck describes the odor as quite fragrant at first but states
that it is lost after a few weeks; I did not notice any especial
odor for my collection. T. albido-brunnea may be distinguished
from our other dimidiate and reflexed species of Thelephora by
its even and pale hymenium and thick spongy pileus. Schwei-
nitz confused one collection of this species with T. biennis Fr.,
from the specimen of which in the Fries Herbarium, at Upsala,
it is clearly distinct. The types of Stereum spongiosum Massee,
Curtis, 3582, and Raveriel, 1 732, in Kew Herbarium, have
colored echinulate spores 8-10 x 6-7//, although described by
Massee as " ellipsoidese 6-7 x 4m" without mention of color and
projections of the wall. The type of Thelephora odorifera Peck,
in Coll. N. Y. State, is somewhat bleached or faded but quite
typical.
Specimens examined:
Exsiccati: Ravenel, Fun. Car. IV, 12, the type distribution of T.
Micheneri B. & C; Ell. & Ev., N. Am. Fungi, 1599, and
Fun. Col., 1209, under the name T. Micheneri in both.
Canada: Toronto, /. Dearness (in Lloyd Herb.).
Vermont: Lake Dunmore, E. A. Burt.
New York: Selku-k, C. H. Peck (in Coll. N. Y. State), the
type of T. odorifera Pk.; Alcove, C. L. Shear, 1010, 1163,
1184; Jamesville, L. M. Underwood.
Pennsylvania: Bethlehem, Schweinitz (in Herb. Schw.), the
type, and also the Nos. 627 and 625 of Syn. N. Am. Fungi
under the names respectively of T. biennis and T. laciniata;
Michener (in Curtis Herb., 3582, and also in Kew Herb.,
same number), type of Stereum spongiosum Massee; Trex-
lertown, W. Herbst, 18, and (in Lloyd Herb., 3052).
viz.,
iiiO ANNALS OF THE MISSOURI BOTANICAL GARDEN
North Carolina: Blowing Rock, G. F. Atkinson, 4322.
South Carolina: Ravenel, 790 (in Curtis Herb, and in Kew
Herb.), under the name Thelephora biennis; Santee Canal,
Ravenel, 1732 (in Curtis Herb, and in Kew Herb.), type of
Stereum spongiosum Massee.
Louisiana : Bogalusa, C. J, Humphrey, 466.
Ohio: Cincinnati, A. P. Morgan, Lloyd Herb., 2627.
Michigan: Saugatuck, E. A. and S. A. Harper, 654.
Wisconsin: Milwaukee Co., comm. by Mrs. F. W. Patterson.
15. T. lutosa Sch w. Trans. Am. Phil. Soc. N. S. 4 : 166. 1834. *
Type: in Herb. Schweinitz.
Pilei cespitose, densely imbricated, at first somewhat fleshy
but at length hard, undulate-phcate, yellowish, almost sub-
tomentose with pulverulence, somewhat horizontally attenu-
ated behind, margin sublobate, at length inflexed; pilcus less
than 2 mm. thick, with hyphae 3/i in diameter; hymenium be-
coming yellowish, even; spores oHve-buff under the microscope,
angular, 5-6 x 3|-4/t.
Cluster about 1^ cm. high and broad.
On the ground in roads and in woods. North Carohna.
_ The type is distinct from T. alhido-hrunnea , having thinner
pileus, finer hyphae, and smaller and paler spores. The pilei
were crowded together into a small buff-colored cluster about
li cm. high and broad, somewhat as in Tremellodendron palli-
dum (Schw.); I failed to find stems at their bases.
Specimens examined:
North Carolina: Salem, Schweinitz (in Herb. Schw.), type.
16. T. cuticularis Berk. Hooker's Lond. Jour. Bot. 6: 324.
1847. Repubhshed in Lea, Catalogue of Plants in Vicinity of
Cincinnati 66. d
Plate 5. fig. 14.
Type: in Kew Herb., and a portion of it from Berkeley „.
Curtis Herb.
Pilei coriaceous-soft, effuso-reflexed or dimidiate, imbricate,
sometimes laterally confluent, drying pinkish buff to cinnamon-
brown, with a broad, pale margin, surface radiately rugose,
soft, silky fibrillosc; substance of the same color as pileus; hy-
menium inferior, concave, even, drab to brownish drab; spores
umbrmous under the microscope, flattened on one side or some-
what kidney-shaped, not angular, echinulate, 8-9 x 6-7/x.
* A figure will be given in Part II.
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 217
Pileus 1-1^ cm. long, 2-4 cm. broad, 1 mm. thick.
On mossy bark at the base of trees and on fallen twigs in
groves. Vermont to Texas and west to Missouri. June to
August.
In his description Berkeley noted that the odor of this species
is strong and unpleasant; my specimens retained such an odor
for several years but I did not notice it before they were dried.
T. cuticularis may be distinguished from our other species by
its drab hymenium, portions of which become sage-green when
crushed under a cover glass in a 7 per cent solution of potassium
hydrate, and by its spores, which are not at all angular or irreg-
ular as regards the main body of the spore, but ovoid and flat-
tened on one side or slightly kidney-shaped and sparingly studded
with slender spines.
Specimens examined:
Vermont: Middlebury, E. A. Burt.
Rhode Island: Olney, 1851 (in Kew Herb, and in Curtis Herb.).
Pennsylvania: Bethlehem, Schweinitz (in Herb. Schw,); the Nos.
628 and 629 of Syn. N* Am- Fungi, under the names respec-
tively of T. fuscocinerea^ and T. gausapata; Kitanning, D.
R. Sumstinej L
Delaware: Newark, H, S. Jackson.
North Carolina: Asheville, H. C. Beardslee^ 03195.
Florida: Mrs. Sams, comm. by C. G. Lloyd.
Texas: W. H. Long, Jr., 351, 387 (in Cornell Univ. Herb.).
Ohio : Waynesville, T. G. Lea (in Kew Herb.), type; Preston, A.P.
and L. V. Morgan, comm. by C. G. Lloyd, also C. G. Lloyd,
specimen dated July 26, 1896; Cincinnati, C. G. Lloyd, 4492.
Wisconsin: Blue Mounds, E. T. and S. A. Harper, 861.
Missouri: Columbia, B. M. Duggar, 289.
17. T. intybacea Pers. ex Fries, Syst. Myc. i : 431. 1821.
Plate 5. fig. 11.
T. intybacea Pers. Syn. Fung. 567. 1801-1807; Myc. Eur.
i:110. 1822.
Illustrations : BuUiard, Champ, de la France pi. ;^75.-Bigeard
et Guillemin, Champ. Super. France 436. pi. 44- /• ^•
Fructifications cespitose, soft, whitish, then rufous-ferru-
ginous, drying chestnut-brown to Rood's brown, with stems
[Vol. 1
218 ANNALS OP THE MISSOURI BOTANICAL GARDEN
somewhat lateral and growing into one another ; pilei imbricated,
fibrous, usually with the fibers matted and agglutinated into
appressed and wholly adnate squamules, margin dilated and
whitish-fimbriate at first, at length becoming entire and colored
like the rest of the pileus; hymenium inferior, concolorous with
the upper surface, papillose; spores concolorous with hyme-
nium, snuff-brown under the microscope, angular-tuberculate,
7-9 X 6-8/x.
Clusters often 5-8 cm. in diameter; individual pileus 2-3 cm.
long, 2-4 cm. broad, 1 mm. thick.
On the ground in pine woods, growing up from the layer of
fallen leaves. Ontario to North Carolina and westward to
Ohio and Michigan. August to October.
The clusters are sometimes central but more often with the
pilei lateral and triangular; sometimes the mass ascends small
sticks and then extends out from this support in reflexed forms;
the upper surface is usually uneven and dries somewhat depressed
between the adnate squamules. This species is distinguished
from ferruginous specimens of T. terrestris by the thicker and
entire margin of the pileus and by the absence of free squamules.
Specimens examined:
Exsiccati: Ell. & Ev., Fun. Col, 1410.
Austria: G. Bresadola.
Ontario: Toronto, /. Dearness, comm. by C. G. Lloyd; Harraby,
Lake Rosseau, E. T. and S. A. Harper, 682.
Maine: Portage, L. W. Riddle, 3.
New Hampshire: Shelburne, W. G, Farlow.
Vermont: Middlebury, Sudbury, Grand View Mt., E. A. Burt
Massachusetts: A. P. D. Piguet, comm. by Dr. Farlow; Natick,
G. E. Morris, No. E.
Connecticut: East Hartford, C. C. Hanmer, 1434-
New York: Alcove, C. L. Shear, 1009; East Gal way, E. A. Burt;
Ithaca, G. F. AtJcinson, Cornell Univ. Herb., 3050, 19652.
Dist. of Columbia: Takoma Park, C. L. Shear, 799, 796; Wash-
ington, 0. F. Cook, 4, comm. by P. L. Ricker.
North Carolina: Asheville, H. C. Beardslee, 0341.
Ohio: A. P. Morgan (in Lloyd Herb.).
Michigan: C. G. Lloyd, 4546; Lawton, L. A. Hawkins; Sailor's
Encampment, Allen and Stuntz, 1, Univ. of Wis. Herb.
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 219
i8. T. terrestris Ehrh. ex Fries, Syst. Myc. i: 431. 1822.
Plate 5. fig. 10.
T. terrestris Ehrh. Crypt. Exsicc. No. 178. 1785.-Persoon,
Syn. Fung. 566. 1801; Myc. Eur. i : 113. 1S22. -Stereum lacin-
iatum Pers. Obs. Myc. i: 36. 179Q.-Thelephora laciniata Feis.
Syn. Fung. 567. 1801.-7". caryophyllea /? laciniata Pers. Myc.
Eur. 1 : 112. 1822.- T. laciniata Fries, Syst. Myc. i : 431. 1821.
Illustrations: Batsch, Elenchus Fung. pi. 24- f- 121.-'Nees,
System der Pilze pi 3^, /. ^5i .-Bolton, Hist. Fung, pi 173.-
Sowerby, Col. Fig. of Eng. Fungi pi 213. -Qooke, Handbook
i: 310.-Stevenson, Brit. Hym. 2: 261.-Smith, Brit. Basid.
399. /. 96 C-E.
Fructifications dark fuscous to fawn-color, coriaceous-soft,
cespitose, obconic, with a short stem-like base, or dimidiate and
sessile, or incrusting and eff uso-reflexed ; pileoli more or less
imbricated, sometimes laterally confluent, fibrous-squamulose
and usually strigose, thin, margin fibrous-fimbriate and lacin-
iate ; hymenium inferior, papillose, fuscous to fawn-color ; spores
pale fuscous, irregular, angular, sometimes slightly tuberculate,
6-9 x 6/i.
Clusters 5-8 cm. in diameter, with single pileolus about 3 cm.
long and broad; obconic pileus 2-3 cm. in diameter; dimidiate
pileolus lf-2 cm. long, 2-3 cm. broad, about 1 mm. thick.
On sandy ground in bare fields and at base of trunks and
from fallen twigs and leaves in pine woods. Canada to South
CaroHna, and in Michigan, Jamaica, and Alaska. July to De-
cember.
My observations of this species acquired from specimens
received and from seeing it growing abundantly near Middle
Grove, N. Y., seem to show that the medium from which this
fungus derives its food produces an interesting effect on the
fructification. Growing from bare, sandy ground the fructifica-
tions are dark fuscous in color, and may be flattened clusters of
imbricated pileoli, or of the obconic-pileus type composed of
ascending pileoli confluent laterally, or dimidiate, sessile pileoli.
When growing on abundant woody matter, as is the case in the
specimen in Sowerby's illustration already cited, the fructifica-
tion assumes a redder color and replaces its dimidiate, sessile
pileus on earth by a reflexed one on the wood. With regard to
220 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
other forms of the clusters and pileoli, the covering of the upper
surface, and the spore characters there is no difference between
those fructifications produced without woody food and those
having it. There is no sharp color separation between these
color extremes.
Specimens growing on the ground usually have a short stem-
like base, while those growing on wood are reflexed; the same col-
lection may show both these conditions, as, for example, that
from Skagway, Alaska, if some of the fructifications start from
sticks and others directly from the ground. Persoon regarded
the stem in T. terrestris as the chief character separating that
species from his T. laciniata, as may be seen from his own de-
5G6 and 567. as folio
the two in his 'Synopsis Fungorum
a
3. Thel, terrestris: subimbricata obscure fusca, pileo applanato
fibrose- strigoso,"
''Hab. m arenosis ad terram. Stipes breuis, lateralis omnino adest.
Substantia submollis, non ita coriacea sicca, vti in ceteris speciebus."
((
^ 4. Thel. laciniata: imbricata obscure fusca, pileo tenui laciniato
crispo subtus papillis congestis scabro."
"Hab. ad radices truncorum. Cespitem difformem efformat, 2 vnc.
lata, tenuis. Stip. vix adest distinctus."
These descriptions supplement each other as a description
for one species; each has special application to fructifications
growing side by side under such conditions as to show that they
are from a common mycelium. Persoon never claimed that his
differed from
Fries gave a different
description of T. laciniata in his works cited — to the injury of
T. intybacea—, but the characters he gives are not satisfactory.
European mycologists with a wide knowledge of the Thele-
phoraceoB as they grow are unable to distinguish these two species.
In letters to me, Brosadola regards T. laciniata as a synonym
of T. terrestris; and Romell does not know T. terrestris if it is
distinct from T. laciniata.
Specimens examined:
Exsiccati: Ellis, N. Am. Fungi, 5U; Ell. & Ev., N. Am. Fungi,
2732, under the name T. intybacea.
Austria: G. Bresadola.
Sweden: G. Romell, 52, 55, 56, 57.
Newfoundland: A. C. Waghorne, 276 (in Mo. Bot. Card. Herb.).
1914]
BURT — THELEPH0RACE2B OF NORTH AMERICA. I 221
Quebec: Gaspe, /. Macoun, 229.
Ontario: Ottawa and Belleville, J. Macoun.
Maine : Wells, J. Blake, comm. by P. L. Ricker.
New Hampshire : Chocorua, W. G. Farlow.
Massachusetts: Magnolia and Woods Hole, W. G. Farlow; Ips-
wich, G. E. Morris, No. F.
Connecticut: South Windsor, East Hartford, and Rockville,
C. C. Hanmer, 1227-29, 9U, 1057.
New York: East Gal way and Middle Grove, E. A. Burt, three
collections from the latter station; Ithaca, G. F. Atkinson,
Cornell Univ. Herb., 22976.
New Jersey: Belleplain, C. L. Shear, 1246; Newfield, /. B. Ellis,
Ellis, N. Am. Fungi, 511.
Pennsylvania: Schweinitz (in Herb. Schw.), the 624 of Syn.
N. Am. Fungi.
North Carolina : Asheville, H. C. Beardslee, 02280; Salem, Schwei-
nitz (in Herb. Schw.), the 624 of Syn. N. Am. Fungi.
Alabama: Tuskegee, Beaumont, 199 (in Curtis Herb.).
South CaroUna: Society Hill, M. A. Curtis, 2693 (in Curtis
Herb.).
Michigan: Agricultural College, G. H. Hicks, Ell. & Ev., Fun.
Col., 2732.
Alaska: Skagway, J. Macoun, Iff; Evans, J^IO (in Mo. Bot.
Gard. Herb.).
Jamaica: Cinchona, W. A. and E. L. Murrill, New York Bot.
Gard., Fun. of Jamaica, 451.
19. T. griseozonata Cooke, Grevillea 19: 104. 1891.
Plate 5. fig. 12.
Tyi^e: in Ravenel, Fun. Amer., 444.
Fructifications cespitose, coriaceous-soft ; pileoli extended into
a short sublateral stem, imbricate, applanate, silky-strigose,
zonate with alternating cervine (Rood's brown) and light buff
zones, margin subfimbriate; hymenium inferior, castaneous
when fresh, drying Rood's brown, rugose, somewhat papillose;
spores pale fuscous, angular, 6-9 x 6-7/i.
Cluster 3-6 cm. in diameter; obconic pileus and single pileo-
lus each 2-3 cm. in diameter.
s
[Vol. 1
222 ANNALS OF THE MISSOURI BOTANICAL GARDEN
On sandy ground in pine woods. New Jersey to Louisiana.
August to November.
This species is closely related to T. terrestris and has the same
habitat, habit of growth, and spore characters, but is distin-
guished from that species by its zonate pileus. The fructifica-
tions usually occur in flattened clusters with spreading pileoli;
sometimes the individual pileoU acquire an infundibuliform
appearance by the growing together for part of their length of
opposite edges of individual pileoli; sometimes a small obconic
pileus occurs composed of two or more pileoli with adjacent
edges confluent. In the collection cited below from Mississippi,
small lobes are present in the cavity of the cup, as in T. vialis
and T. caryophyllea.
Specimens examined:
Exsiccati: Ravenel, Fungi Am., 444, type distribution; Ravenel,
Fun. Car, II, 28, under the name T. caryophyllea; Ellis, N.
Am. Fungi, 714; Ell. & Ev., Fun. Col., 1305.
New Jersey: Newfield, J. B. Ellis, in his exsiccati cited.
South Carolina: Aiken, //. W. Ravenel, Fungi Am., 444, type
collection.
Alabama: Auburn, C. F. Baker, Lloyd Herb., 3462.
Mississippi: Biloxi, Mrs. E. S. Earle, 32.
Louisiana: St. Martinville, A. B. Langlois, by.
20. T. fimbriata Schw. ex Schweinitz, Trans. Am. Phil. Soc.
N. S. 4: 166. 1834.
Plate 4. fig. 3.
Merisma fimhriatum Schw. (Syn. Fung. Car., No. 1067)
Schrift. d. Naturforsch. GeselL, Leipzig, i: 110. 1S22 -Thele-
phora scoparia Peck, Rep. N. Y. State Mus. 42: 123 (27). pi.
2. f. 20, 21. 1889.
Illustrations: Peck, Rep. N. Y. State Mus. 42: pi. 2. f. 20,
21.
Type: in Herb. Schweinitz.
Fructification coriaceous-soft, incrusting and ascending small
plants (mosses, etc.), here and there emitting fascicles of
branches united below, subterete, acuminate or fimbriately
incised, at first pale or whitish, soon ferruginous brown, dry-
ing Rood's brown; hymenium even, pruinose-pubescent; spores
umbrinouSj tuberculate, 7-11 x 6-9 n.
19U]
BURT — THELEPHORACE^ OF NORTH AMERICA. I 223
Incrusting and ascending upward 1-3 cm.; free branches 5-10
mm. long, 1 mm. thick, sweep of fascicle about 5-10 mm.
In moist places. New York to South Carolina, and west to
Illinois. July and August.
The type is an incrusting specimen, covering as its main axis
a small twig in one specimen and a moss in the other, and send-
ing out a few lateral branches which are flattened towards the
free ends and subfimbriate ; main trunk is cylindric, latericius
(of 'Chromotaxia'), ends of branches paler; spores umbrinous
under the microscope, tuberculate, 7-8 x 6 /i. Schweinitz de-
scribed the species as becoming hard and cartilaginous, but
this is an error probably due to the foreign matter surrounded
by the main trunk. Several other specimens are present in
his herbarium under various names.
Specimens examined:
Exsiccati: Ellis, N. Am. Fungi, 512, under the name T. cristata.
Massachusetts: Weston, A. B. Seymour, T 1 (in Mo. Bot.
Gard. Herb., 45573).
New York: Bethlehem and Selkirk, C. H. Peck (in Coll. N. Y.
State), type of T. scoparia; Syracuse, from Herb. Cornell
Univ., 19474.
New Jersey: Newfield, J. B. Ellis, N. Am. Fungi, 512.
Pennsylvania: Bethlehem, Schweinitz (in Herb. Schw.), the 615
of Syn. N. Am. Fungi, under the name T. stahularis.
North Carolina: Salem, Schweinitz (in Herb. Schw.), type, and
also the 1063 of Syn. Fung. Car., under the name Merisma
fuscescens.
Indiana: Millers, E. T. and S. A. Harper, 670.
Illinois: Havana, H. C. Beardslee; Riverside, E. T. and S. A.
Harper, 668.
21. T. perplexa Burt, n. sp.i
Type: in Curtis Herb.
Fructification incrusting, coriaceous, consisting of a resupinate
membrane from the central portion of which arise cylindric
trunks either simple or digitately branched; resupinate portion
spongy, firm, separable, fuscous at the center, margin thin,
determinate, pinkish buff; ascending portions spongy, firm,
1 A figure will be given in Part II.
[Vol. 1
224 ANNALS OF THE MISSOURI BOTANICAL GARDEN
fuscous, simple and tapering upward or soon branching and
terminating in paler either subulate tips or somewhat flattened
€nds; spores fuscous, subglobose, echinulate, 8-10 x 8-9 /i.
The resupinate membrane may be 3 cm. in diameter ; ascending
portion of fructification 2-3 cm. high, l|-2 mm. thick.
On decaying leaves and sticks on the ground. Cuba.
Berkeley & Curtis based their description of Thelephora den-
tosa on two collections made in Cuba by C. Wright in different
years; these collections are different specifically. The original
description applies chiefly to the earlier collection, made in
1857, which is unnumbered. I take my type of T. perplexa
from the later collection, C. Wright, 238.
Specimens examined:
Exsiccati: Fungi Cubenses Wrightiani, C. Wright, 238, under
the name Thelephora dentosa B. & C.
Cuba: C. Wright, 238, type (in Curtis Herb.).
22. T. dentosa Berk. & Curtis emend Burt.^
T. dentosa B. & C. (Fungi Cubenses) Jour. Linn. Soc. Bot.
10 : 329. 1867.
Type: type and cotype in Kew Herb, and Curtis Herb, re-
spectively.
Fructification coriaceous-soft, incrusting leaves and small
twigs on the ground and ascending as free, sessile, dilated, tri-
angular, flabelliform pilei which are dentate at the upper end
or deeply divided into a few finger-shaped divisions, honey-
yellow to tawny olivaceous throughout, minutely hairy under
a lens; spores honey-yellow, globose to ovoid, weakly echinulate,
6-10 X 6-8 M.
Pileus 1 cm. high, 5 mm.-l cm. broad.
On rotten vegetation. Cuba. June.
As already stated in connection with T. perplexa, Berkeley
& Curtis cited for types of their T. dentosa specimens from two
collections made in Cuba by C. Wright. These collections were
made with an interval of several years between the collections,
which differ specifically. As noted by Berkeley & Curtis, their
description applies better to the earlier collection, to which I now
* A figure will be given in Part II.
1914]
BURT — THELEPHORACEiB OF NORTH AMERICA. I 225
restrict their species. This earlier collection was distributed
by C. Wright; unnumbered, under the name Thelephora dentosa
B. & C. before the publication of the description of this species,
and the cotype in Curtis Herb, is unnumbered also. By what
was apparently a slip of the pen, Berkeley cited this type as
C. Wright, 507. By the kindness of Dr. Farlow I have been
permitted to examine the manuscript records which show that
Wright collected only one No. 507, which was determined by
Berkeley as Xylaria ohovata Berk, and is cited under this species
by Berk. & Curtis, Jour. Linn. Soc. Bot. lo: 380. 1867. I
find in Curtis Herb, such a specimen labelled Xylaria ohovata
Berk., Cuba, C. IVrightj 507. I conclude that the type and
cotype of T. dentosa B. & C, first cited In their description,
are from the collection distributed by C. Wright, unnumbered,
under the name Thelephora dentosa B. & C.
Specimens examined:
Exsiccati: Plantae Cubenses Wrightianae, unnumbered, under
the name Thelephora dentosa B. & C.
Cuba: C. Wright, cotype (in Curtis Herb.).
23. T. spiculosa Fries, Syst. Myc. i: 434. 1821; Epicr. Syst.
Myc. 539. 1836-38. Plate 4. fig. 2.
Illustrations: Persoon, Syn. Fung. pi. 3. f. 16.
Type : an authentic specimen from Fries, in Kew Herb.
Fructifications cespitose, from byssoid becoming fleshy, vari-
able by incrusting habit, pale buff at first, main portions
becoming purplish-fuscous (Rood's brown) with age, ramose-
spiculous, tips penicillate and whitish; spores umbrinous under
the microscope, irregular, echinulate, 8-9 x 6-7 ju.
Clusters 1-2 cm. high, 2-4 cm. in diameter, single fructifica-
tion 1-2 cm. high, about 1 mm. in diameter, with branches
spreading 4-6 mm.
On leaves on ground in moist groves. Ohio to Wisconsin.
July. Rare.
The best specimens which I have seen have main trunks of
the fructifications running side by side over partially decayed
beech leaves and confluent into an effused mass. These trunks
ascend obHquely from the leaves to a height of 1-2 cm., branch
sparingly, and terminate in spiculous tips. The fructification
[Vol. 1, 1914]
226 ANNALS OF THE MISSOURI BOTANICAL GARDEN
must be inconspicuous in the woods since tlie general color of
the mass is the same as that of the leaves on which it is effused,
although the main trunks may be darker.
Specimens examined :
Exsiccati: Kunze, Fun. Sel. Exsic, 560.
Sweden: specimen from Fries (in Kew Herb.).
Austria: G. Brcsadola.
Ohio : Preston, C. G. Lloyd.
Michigan : Glen Lake, C. G. Lloyd, 02^71 .
Wisconsin: Lake Geneva, E. T. and S. A. Harper, 883.
(To be continued.)
Explanation of Plate
PLATE 4
All figures of plates 4 and 5 have been reproduced natural size from
photographs of dri(;d herbarium specimens of species of Thelephora.
Fig. 1. Thelephora anthocephala. From specimen collected at Lin wood, Ohio, by
C. G. Lloyd, No. 02164.
Fig. 2. T. spiculosa, a, from specimen on leaves of Fagus collected in Europe by
Bresadola, which I compared with the specimen from Fries in Kew Herbarium;
6, from specimen collected at Glen Lake, Mich., by C. G. Lloyd, No. 02471.
Fig. 3. T, Jimbriata. From specimen incrusting living strawberry {Fragaria)
plant, collected at Riverside, 111., by E. T. and S. A. Harper, No. 668.
palmat
4612.
Fig. 5. T. magnispora. From type specimens collected at Chester Vale, Jamaica,
y W. A. and Edna L. Murrill, No. 295. a shows upper surface and side of
ileus, and 6, the hymenium.
Fig. 6. T. regularis. From a sketch of the type in Herb. Schwelnitz.
Fig. 7 a. T. multiparlita. From specimens collected at Trexlertown, Pa., by Dr,
W
Fig. 7 b. T. regularis. From specimens collected at Clayton, Del., by H. S. Jack-
son.
Wa^h
Suksdorf
Fig. 9. T. caryophyllea. From specimens collected in Michigan, by C. G. Lloyd,
No. 4547.
\
\
V
Ann. Mo. Bot. Gard_,Vol. 1, 1914
Plate 4
r
\
)
*1
^
i
b
BLTRT—THELEPHORACEAE OF NORTH AMERICA
1. THKLEPHORA ANTHOCEPHALA.— 2. T. SPICUL0SA.--3. T. FIMBRIATA.
4.T. PALMATA. — 5. T. MAGNISPORA.— 6 and 7 b. T. REGULARIS.— 7 a. T. MULTIPARTITA,
— 8. T, SCISSILIS — 9. T. CARYOPHyLLEA.
COCKAYNE. BOSTON.
228
[Vol. 1, 1914
ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 5.
Middle Grove, N. Y.
terrestris. From specimens collected on ground in open
fimbriate
Fig. 11. r. intybacea.
ne leaves and twies
hym
woods incrusti
r surface with
adnate squamules and whitish, thick, entire margin; h, the hymenium
fiurface
griseozonata
Fun. Amer.. No. 444
specimen of type collection, distributed in Ravenel
hrunnea
upper side of specimen collected
and S. A. Harpe
The specimen is about
hymenium of specimen
cuticvlaris. From specimens collected at Blue Mounds, Wis
tmder
and S. A. Harpe
viewed obliquely from above: 6. viewed
vialis.
collected at Lake Dunmi
^
c
AxN. Mo. BoT. Gard., Vol. 1. 1914
Plate 5
b
BURT — THELEPHORACEAE OF NORTH AMERICA
10. THELEPIIORA TERRESTRIS. — U. T. INTYBACEA.— 12. T. GRISEO-ZONATA
13. T. xYLBIOO-BRUNNEA. — 14. T. CUTICULARIS. — 15. T. VIALIS.
i
I
COCKAYNE. BOSTON.
INDICATIONS REGARDING THE SOURCE OF COM-
BINED NITROGEN FOR ULVA LACTUCA
G. L. FOSTER
Teaching Fellow in the Henry Shaw School of Botany of
Washington University
Introduction
Very little attention has been given the question of the
sources of nitrogen for marine algse. Nevertheless, the ques-
tion is an interesting one both physiologically and ecologically,
because of the extremely small amount of nitrogen supposed
to be present in sea-water, and because of the very noticeable
change in the type of algal flora when the nitrogen content of
the environment is increased, as by the presence of sewage.
The literature bearing on the subject is practically limited to
a debate between a few authors as to the amount and form
of nitrogen in sea-water, and the way in which the supply is
maintained. This dispute involves some questions of funda-
mental importance for marine biology; consequently, a brief
statement of the different views is pertinent.
Natterer (13) reports that careful analyses of water from
the high seas show scarcely a trace of nitrates. Nitrites are
somewhat more abundant, but not sufficiently so to admit
of quantitative determination. Ammonium compounds, on the
other hand, according to Thoulet, are present in sufficient
amount to be quantitatively determined, and vary from .13
to .34 mg. per liter (.013-.034 per cent) according to the
locality. Reinke (15) considers these amounts of nitrogen
reported to be insufficient for the production of the enormous
amount of living material in the sea, especially when the activity
of nitrifying and denitrifying bacteria is taken into account.
He considers as of prime importance in this question the nitro-
gen-fixing bacteria which have been demonstrated in sea-water
by Benecke and Keutner (4), and others. Reinke found Azo-
tohader embedded in the gelatinous material on the surface
of Laminaria fronds and argues for a symbiotic relation between
the algse and bacteria.
Akn. Mo. Bot. Gabd., Vol. 1, 1914
(229)
[Vol. 1
230 ANNALS OF THE MISSOFIU BOTANICAL GARDEN
Brandt (7), however, attaches Uttle or no importance to
Reinke's view, and maintains that the nitrogen content
of sea- water is determined by a balance between the activity
of denitrifying bacteria, on the one hand, and the great amount
of nitrogenous material carried to the sea by the rivers, on the
other. Brandt (5, G) considers that the nitrogen content of
sea- water is at a "minimum" and is the limiting factor in tlie
production of marine organisms. Considering especially the
plankton life, he finds that the amount of plankton is propor-
tional to the nitrogen content of the water, and correlates the
comparative poverty of tropical seas in plankton life with the
relatively greater activity of denitrifying bacteria in the warmer
waters.
More recently Putter (14) has reported that analyses of the
water from the Gulf of Naples give per liter .18 mg. of nitrogen
in nitrates and nitrites, and .56 mg. in ammoniacal nitrogen.
Furthermore, he claims that these figures represent less than
half the total combined nitrogen actually present in sea-water.
In his opinion there is no need for considering the nitrogen
content to be at a "minimum" since it is present in greater
concentration than the carbon dioxide. It is impossible to
say which of these views is the correct one, and further work
in this field is much needed.
The above named authors incidentally assume that nitrogen
is available for the algae only in the form of nitrates or ammo-
nium salts. This is entirely an a priori assumption, as no data
are offered in support of such a view. On the contrary, it
seems more likely that the alga) can use many organic ni-
trogen compounds. This would seem probable in the light of
recent work which has been done on the fresh-water algae.
In regard to the nutrition of the fresh-water forms we have
departed far from the old idea that green plants are strictly
autotrophic. Thus, by the work of Beyerinck (3), Charpentier
(8), Chick (9), Artari (1, 2), and others, it has been established
that many of the fresh-water algae have, with respect to nitrogen,
distinct saprophytic tendencies,— preferring organic to inor-
ganic nitrogen. Artari, especially, has shown that several
algae {Chlamydornonas, Stichococcus , ChloreUa, Scenedesmus, and
others) can grow and retain the chlorophyll under completely
others, ammo
1914] „ql
FOSTER— SOURCE OF COMBINED NITROGEN FOR ULVA ZUL
saprophytic conditions, as in solutions containing amino acids
and glucose in the absence of light and carbon dioxide. In all
these cases, however, growth is more rapid under so-called
mixotrophic conditions, i. e., with both organic nitrogen and
carbon present in addition to sunlight and carbon dioxide.
Artari takes up the question of the relative value of different
nitrogenous compounds, and shows that they vary greaUy
with different algae,— some .
acids and ammonium salts, and a few, nitrates. On the whole,
the majority of forms investigated grow best in the presence
of amino nitrogen. Many alga, especially of this last class,
are often found in water polluted with sewage or decaymg
organic matter.
Among the marine algse, there is a more or less definite flora
characteristic of sewage-polluted waters. Most conspicuous
among the plants of this group are the species of Ulva. Letts
and Richards (11), in their reports on sewage in British har-
bors, state that Ulva latissima grows in excessive quantities
in polluted waters, and they find that the nitrogen content of
this seaweed varies with the degree of pollution of the water.
Cultural experiments conducted by Letts and Richards showed
that Ulva latissima grows more rapidly in a mixture of sewage
and sea-water than in pure sea-water alone.
Experimental
Preliminary experiments were made at the Woods Hole
Laboratory to determine the sources of available nitrogen for
Ulva lactuca. The algal material used in the experiments was
collected at the mouth of an inlet where the water was at all
times highly polluted with
tained in the laboratory in glass tumblers containing 150 cc.
of solution. When brought in, the fronds of Ulva were well
rinsed in clean sea-water and cut into strips exactly 3 cm
main
C
length and about 2 cm. wide. Three such strips were placed
each vessel, and the cultures kept at a temperature of 21
by placing the vessels in a tray of running water. In each
case the solution was renewed at the end of 5 days. After 10
days the strips were again measured and the increase in length
recorded.
^6^ ANNALS OF THE MISSOURI BOTANICAL GARDEN
^^ Two main types of nutrient solution were used,— one (solu-
A) being natural sea-water, the other (solution B) being
artificial
These stock sea-waters
non-
were made double strength and subsequently diluted by the
addition of distilled water and the stock solution of the nitro-
genous compounds to be tested. The following nitrogen com-
pounds were used in the experiments: ammonium nitrate,
urea, acetamid, sodium asparaginate, acetanilid, and dimethyl-
anilin. Parallel experiments were run, adding these compounds
to solution A and solution B.
Preliminary tests roughly determined the maximum
toxic concentrations of these compounds when added t
water to be:
Ammonium nitrate 0.011 gram molecular
^^^^ . 010 gram molecular
Acetamid 0.250 gram molecular
Asparagin 0.080 gram molecular
The table presents the results of the experiments. All figures
for concentrations represent fractions of gram molecules per
liter, except in the case of dimethylanilin. Here the solubiUty
was not known and the figures represent fractions of a saturated
solution in distilled water at 20«C. In the column headed
tt i-L.)}
growth" is recorded the increase in length in millimeters of
the strips of Ulva after 10 days in the solution. In each case
the figures for growth represent the average o^ three or more
cultures. Checks show the growth in solutions A and B with
no additional nitrogen.
^ It is apparent from the following table that, under the condi-
tions of the experiment, ammonium nitrate and urea are consider-
ably better nutrients for Ulva than the other compounds used.
These two cause a marked increase in growth over that of the
controls, in both the artificial and natural sea-waters. The
nutritive value of these compounds was also indicated by the
healthy appearance of the cultures. The alg£e were of a deep
green color, very turgid, and considerably curled by rapid
growth. Judging from the growth and general appearance of
the cultures, there is httle choice between the nutrient values
of ammonium nitrate and urea.
1914]
FOSTER — SOURCE OF COMBINED NITROGEN FOR ULVA
233
COMPARATIVE TABLE SHOWING GROWTH OF STRIPS OF ULVA IN VARIOUS
NITROGEN-CONTAINING SOLUTIONS
Ammonium nitrate
Urea
Acetamid
/^
Growth
-
1. ^
Cone.
Growth
Cone.
Growth
Cone,
Sol*
A
Sol.*
B
0.3
Sol.*
A
0.8
Sol.*
B
Sol.*
A
Sol.*
B
Check
0.8
1.0
Check
0.3
Check
0.8
0.8
1.0
0.3
0.00005
0.5
0.0005
1.4
0.5
0.001
0.4
. 0001
1.4
1.5
0.001
0.005
1.6
0.6
0.005
0.4
0.0005
1.9 2.0
1.6
1.4
0.01
0.7 0.5
0.001
1.4
1.2
0.01
0.9
1.3
1
0.10
1.0 1.0
Sodium as
paraginate
1
1
Acet
1
anilid
Dimethylanilin
y^
Growth
Cone.
Gro
Sol.*
A
0.8
iwth
Conc.f
Growth
Cone.
1
Sol.*
A
Sol.*
B
Sol.*
B
Sol.*
A
Sol*
B
Check
0.8
0.3
Check
0.3
Check
0.8
0.3
0.002
0.7
0.6
0.0005
0.6
0.5
0.0
0.0
0.002
0.3
0.4
0.01
0.9
0.2
0.0025
0.0
0.0
0.02
0.0
0.5
0.05
0.7
0.0
0.0125
0.10
0.0
0.0
*Sol. A= natural sea-water; sol. B = artificial sea-water.
t Cone, under dimethylanilin represents fractions of a sa
tilled water at 20*^0.
Acetamid has a somewhat lower nutrient value than ammo-
nium nitrate or urea, but still it causes a greater growth than
do the control solutions to which no foreign nitrogen was added.
The alga in acetamid solutions appeared normal in every way.
The results with the sodium asparaginate were rather unex-
pected. This compound is well known to be a good nutrient
for many fungi and fresh-water algae. For Ulva, on the other
hand, sodium asparaginate appears to have no appreciable
nutrient value. In no case did it cause any notable increase
in growth, although the algal material appeared perfectly normal.
[Vol. 1
234 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Acetanilid and dimethylanilin are in a separate class, — being
decidedly toxic at all the concentrations used. At the lowest
concentrations there was slight growth at first, but in ten days
all cultures were dead and discolored. The results with these
last two compounds are comparable to those obtained with
similar substances by Czapek (10) and by Lutz (12) working
on fungi and fresh- water algse. They found that compounds
having the nitrogen attached directly to a benzene nucleus are
toxic.
Pure culture methods were not attempted on account of the
brief time available for this work, and the question of the pos-
sible interaction of ammonifying bacteria is therefore pertinent.
However, the rapid augmentation of growth upon the addition
of the amido compounds, and the comparative absence of
bacteria both suggest a direct absorption of these substances.
Moreover, since rapid growth of the alga occurs in concentra-
tions of the amido compounds considerably greater than the
toxic limit for ammonium salts, and since, further, no evi-
dence of toxicity of fairly strong solutions of urea and aceta-
mid developed during the interval of these experiments, no
support is given to the thought that ammonification may be
an important factor. However, in further continuation of this
work it is proposed to control this possibility by quantitative
tests.
It seems probable from the facts brought out here, as well
as from the work of Letts and Richards, that Ulva is not lim-
ited to an inorganic nitrogen supply, since growth occurs with
urea or acetamid as the sole source of nitrogen, and, as Letts
and Richards have shown, that it grows more rapidly in sewage-
polluted water than in pure sea-water. Undoubtedly, further
experiments would show that other organic compounds can
supply available nitrogen for Ulva.
The results also indicate that for Ulva, at least, the amount
of available nitrogen in the water is the limiting factor in growth.
This is shown by the fact that growth is more rapid in sea-
water containing additional nitrogen (ammonium nitrate, or
urea) than in pure sea- water. The above mentioned results
of Letts and Richards also point to the same conclusion, as does
1914]
FOSTER — SOrRCE OF COMBINED NITROGEN FOR ULVA 235
the abundant growth of Ulva in nature in waters polluted with
sewage.
In conclusion, the writer is pleased to express his thanks for
the generous assistance given during this study by Prof. B*
M. DuggaF; under whose direction the work was carried out
while occupying a research table maintained by Dartmouth
College at the Marine Biological Laboratory, Woods Hole,
Massachusetts.
Graduate Laboratory^ Missouri Botanical Garden.
Literature Cited
1. Artari, A. Zur Frage der physiologischen Raseen einiger griinen Algen. Ber. d.
deut. bot. GeB. 20: 172-75. 1902.
2. Zur Physiologic der Chlamydomonaden. Jahrb. f. wiss. Bot. 52:
410-66. 1913.
andcren
W. Culturvereuche mit Zoochlorellen, Lichenengonidien und
rftn Aken. Bot. Zeit. d8: 725-54, 757-67. 781-85. 1890.
W.. und Keutner, J, tjber stickstoffbindende
333-46. 1903.
Brandt
Meeresunters. N.F. Abt.
215-30. 1899.
6.
7.
im Meere. 2. Abhandl. Ibid. 6: 25-79. 1902
un
Meere, Beih. bot. Centralbl. 16: 383-402. 1904.
Charpentier, P. G. Alimentation azot(5e d'line algui
Ann. Inst. Pasteur 17: 321-34. 369-420. 1903.
9. Chick, H. A study of a unicellular green alga occurring in polluted water, with
special reference to its nitrogenous metabolism. Proc. Roy. Soc. 71: 458-77.
1903.
10. Czapek, Fr. Zur Kenntniss der Stick st off versorgung und Eiweissstoffwechsel
bei Aspergillus niger.
11. Letts, E. A., and Richards, E. H.
130-39. 1902.
issun
in relation to the pollution of the waters in which they occur. Seventh Report
of the Roy. Comm. on Sewage Disposal, Appendix 3: 72-100. 1911.
12. Lutz, L. Sur Taction exerc^e sur les vdg^taux par les composes azot6es organ-
iques a noyau benzenique. Compt. rend. Cong. Soc. Sav. Paris 1903: 65-69.
13. Natterer, K. Chemische Untersuchungen von der Expedition S. M. Schiff
'Tola." Denkschr. k. Akad. Wiss., Wien, math.-naturw. Kl. 65: 445. 1898.
14. Putter, A. Der StofThaushalt des Meeres. Zeitschr. f. allgcm. Physiol. 7:
321-68. 1907.
15. Remke, J. Die zur Emahrung der Meeresorganismen disponiblen Quellcn
an
Ber. d. deut. bot. Ges. 21: 371-80. 1903.
THE EFFECT OF CERTAIN CONDITIONS UPON THE
ACIDITY OF TOMATO FRUITS
B. M. DUGGAE
Physiologist to the Missouri Botanical Garden
Professor of Plant Physiology in the Henry Shaw School of Botany of
Washington University
AND M. C. MERRILL
Research Assistant to the Missouri Botanical Garden
In a recent communication the senior author (4) has referred
to the possibility that the total acid content of tomato fruits
ripened at a temperature of 30 °C., or above, may be related
in some way to the failure of lycopersicin development at that
temperature. It was determined that the ''total acidity for
green, ripening, and ripe fruits, grown under the same condi-
tions, is unexpectedly uniform, amounting to .57 to .58 per
cent citric acid." The fruits just referred to were of the
same variety picked at the same time. The tests of acid con-
tent of incubated fruits were made later in the season, and
these indicated a lower acidity than that of normally green or
ripe fruits. At that time the requisite material was obtained
from the Department of Horticulture, Cornell University.
During the past summer several varieties of tomatoes were
grown in the Missouri Botanical Garden in order to furnish
material for further pigment studies, and incidentally this
material has enabled us to determine with greater care the acid
content of tomato fruits, especially of different varieties, and
likewise the comparative acidity of fruits direct from the field
and of those of the same picking incubated for various intervals.
The tests included below were made by pulping thoroughly
a weighed quantity of the tissue (15 gm.), diluting with 150
cc. distilled water, employing for each titration 25 cc. of this
solution diluted with distilled water to 50 cc, and titrating
with n/10 NaOH, using phenolphthalein as indicator. Not less
than two titrations were made in any case, and these were from
one or more samples of tissue. The accompanying table
Ann. Mo. Bot. Gabd., Votj. 1, 1914
6
(237)
238
ANNALS OF THE MISSOUEI BOTANICAL GARDEN
[Vol. 1
indicates the variety and condition of the fruit; quantities of
n/10 NaOH required to neutralize; and the per cent of acidity
in terms of citric acid.
TABLE SHOWING ACID CONTENT OF TOMATO FRUITS
^V Y
Condition
Average no.
of cc. of
n/10 NaOH,
to neutralize
Total per
Variety
When
picked
Interval
or incubation
When
titrated*
cent of acid
as citric
Dwarf Stone
Dwarf Stone
Dwarf Stone
Dwarf Stone
Dwarf Stone
Ripe
Half grown
Half grown
Half grown
Half grown
Ripe
Half grown
Incub. 32° C. 10 days
Lab. 24 days
Incub. 32** C. 10 days
1
Red
Green
Artif. yellow
Red
Green
1 . 695
1.82
2.135
1.375
1.485
.52
.50
.66
.42
.46
Sparks* Earliana
Sparks* Earliana
■m
Red
Green
1.695
1.87
.52
.58
Truckere' Favorite
Truckers' Favorite
Half grown
Half grown
r
Incub. 32° C. 22 days
Lab. 24 days
Artif. yellow
Red
2.56
1.66
.79
.51
Red Peach
Red Peach
Half grown
Half grown
Incub. 32° C. 22 days
Lab. 24 days
1
Artif. yellow
Red
* 2.115
1.675
.65
,52
1
Yellow Peach
Yellow Peach
Half grown
Half grown
Incub. 32° C. 22 days
Lab. 24 days
1
Artif. yellow
Yellow
2.47
2.065
.76
.64
Yellow Plum
Yellow Plum
Ripe
Half grown
Yellow
Green
2.12
1.92
.65
.59
Yellow Pear
Yellow Pear
Half grown
Half grown
Incub. 32° C. 20 days
Lab. 24 days
Artif. yellow
Yellow
1.60
1.395
.49
.43
♦All fruits designated "red," "yellow," and "artificial yellow" were, at the same time, ripe.
The results above reported may not yet be as extensive as
might be desired in order to follow closely the changes in acid-
ity under different conditions; but they consistently point out
certain relations of interest which may be briefly enumerated
follows: (1) A
the acid content of green
pened fruits was made, using Dwarf Stone
and normally r
Sparks' Earliana, and Yellow Plum, all direct from the field
There were no marked differences between the green and ripe
stages within the variety; yet the acidity of the green fruits
of the red varieties in these tests is somewhat higher, while
the acid content of the green fruits of the one vellow vanptv
tested is somew
(2)
Stone, Truckers
Favorite, Red Peach, Yellow Peach, and Yellow Pear which
1914]
DUGGAE AND MERRILL^ — ACIDITY OF TOMATO FRUITS 239
were picked green and ripened in the incubator at 32-33°C.
(10-22 days) exhibit a higher acid content than either those
ripened on the vines or those ripened at the temperature of the
laboratory. (3) There are considerable differences in the acid-
ity of varieties, but judging from the results of these tests
the normally ripened fruits of yellow varieties commonly con-
tain as much acid as those of red varieties.
The several facts brought out by these tests render it obvious
that there is now no sufficient evidence to justify relating
pigmentation to total acidity. The acidity changes are, how-
ever, interesting in themselves, in these as well as in other
fruits. No attempt was made to follow progressively any
changes in acidity induced by conditions; but in titrating on
one occasion, after an interval of two days, new samples of
both red and yellow fruits which had been ripened in the labo-
ratory, it was found that the acidity had noticeably declined
since the previous titrations from the same lots of fruits.
We have reckoned the acidity of the tomato in terms of
citric acid, as is customary. It should be noted, however,
that while Bowman (3) and others report citric as the chief
acid of the tomato, Albahary (1), on the contrary, gives .48
per cent as the malic acid content and .09 per cent as that of
citric acid in the fresh fruits. The author last mentioned
gives no indications respecting the variety or condition of the
fruit employed. In a later contribution (2) he reports the
results of analyzing tomato fruits in different stages of matura-
tion, as follows: ''1° le fruit vert avant Tapparition de la graine
dans la pulpe; 2° le fruit vert au moment ou la graine est
completement f ornate; 3° le fruit rouge arriv^ k sa pleine matu-
ration." In the second stage, corresponding to practically
full grown, green, he finds .58, and in the ripe fruits .42 per
cent of organic acids. This is in complete agreement with
our findings. In the earliest stage of fruit development Alba-
hary finds an acid content of only .116 per cent. Wehmer
(5), after quoting Albahary (1) as to the percentage of the
various acids in the fruit, remarks, ''Die Aciditat wechselt
stark je nach dem Reifestadium (von 0,06-0,697% des Saftes
auf Citronensaure berechnet)." He does not indicate the
(Vol. 1, 1914]
240
ANNALS OF THE MISSOURI BOTANICAL GARDEN
urce of these data, and certainly the smaller percentage gh
n refer only to the youngest stages of fruit development
Graduate Laboratory^ Missouri Botanical Garden.
Literature Consulted
1. Albahary, J. M. Analyse complete du fruit du Lycopersicum eeculentum ou
Tomate- Compt. rend. acad. Paris 145: 131-33. 1907.
, fitude chimique de la maturation du Lycopersicum esculentum (To-
2.
mate). Compt. rend, acad, Paris 147: 14&-47. 1908.
3. Bowman, W. Tomatoes: chemical examination of fruits. Va. Agr. Exp. Sta.
Bui. 9: 16-18. 1891.
4. Duggar, B. M. Lycopersicin, the red pigment of the tomato, and the effects of
conditions upon its development. Washington Univ. Studies i : 22-45. 1913,
5. Wehmer, C. Die Pflanzenstoffe 685-86. 1911.
A METHOD FOR THE DIFFERENTIAL STAINING OF
FUNGOUS AND HOST CELLS
R. E. VAUGHAN
Assistant in Plant Pathology, University of Wisconsin
Exchange Fellow in the Henry Shaw School of Botany of Washington University
In making histological studies of fungi on living or dead
plant tissues the use of the stain known as ''Pianeze Illb"
has been found very satisfactory in differentiating the fungus
from the plant substratum, this differentiation occurring both
in lignified and unlignified cell walls. The host tissue stains
green and the mycelium a deep pink. This stain, devised
by Dr. Pianeze for the study of cancer tissue,^ is made up as
follows :
Malachite green . 50 gm.
Acid f uchsin 0.10 gm.
"Martius gelb" 0.01 gm.
Water, distilled 150.00 cc.
Alcohol, 95 per cent 50 , 00 cc.
Dr. Pianeze reports that it gives the following staining
reactions: green in chromatin of resting or dividing nucleus,
rose in cell protoplasm and membrane, and red in cancer bodies.
For use with plant tissues the procedure is as follows: Wash
in water or alcohol, stain in the undiluted mixture 15-45
minutes, remove excess stain in water, and decolorize in 95
per cent alcohol to which a few drops of hydrochloric acid have
been added. For permanent mounts, clear with a carbol-
turpentine mixture, remove clearer in xylol, and mount in
balsam. Preparations of Stereum^ Corticiumy and Polystictus
have been made with great success.
This stain is also valuable for staining germinated spores on
the surface of a leaf. The procedure in this case is as follows;
Infect marked portions of a leaf with a suspension of spores
applied with a pipette, and place the plant under suitable
conditions for fungous growth for 24-48 hours. Then permit
^ Pianeze, G.
und
Beitrag^
z. path. Anat. u. z. allg. Path. Supplement i: 1-193, 1896. [cf. p. 58,]
Amr. Mo. BoT. Gaed., Voii. 1, 1914
(241)
[Vol. 1, 19141
242 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the leaf to dry in the air, remove the area desired from the
balance of the leaf, and place in a killing fluid. The best com-
bined killing and tissue-clearing mixture for this purpose is
one recommended by Dr. Duggar, composed of glacial acetic
acid and 95 per cent alcohol. I have used equal parts of these
agents most advantageously. This dissolves the chlorophyll,
renders the leaf transparent or nearly so, and at the same time
fixes the fungus with little plasmolysis. Allow the killing mix-
ture to act for 24-36 hours; wash in 50 or 70 per cent alco-
hol^ to remove the acid; and pass successively through the stain
(15-30 minutes), water (2 minutes), acid alcohol (as short a
F
time as possible), carbol-turpentine (until clear), xylol (until
clearing agent is removed), and then mount in balsam. This
process of differential staining has been successfully used
with Ascochyta Pisi on pea, Helminthosporium sativum on
barley, and Phoma Brassicoe on cabbage.
Pianeze's stain has not given as good results with the rusts
as Durand's combination of Delafield's haematoxylin and eosin.
Durand's stain^ was not uniformly successful, however, and it
was found that one of the chief difficulties often experienced
finds its explanation in the killing solution which the stain
follows. Fleniming's solution, which was first used, gave
very poor results. A modification of Gilson's mercuric chloride
solution was found most satisfactory. This solution, as recom-
mended by Dr. Durand, is made up as follows:
Water, distilled 60 cc.
Alcohol, 95 per cent 42 cc.
Acetic acid, glacial 18 cc.
Nitric acid, concentrated 2 cc.
Mercuric chloride, sat. aq. sol 11 cc.
Diseased tissue may be fixed from 6 to 24 hours, then washed
in 65 per cent alcohol, run through the alcohols, infiltrated
■
with cedar oil, and imbedded in paraffin. This method is
undesirable for nuclear structures, but gives excellent prepara-
tions for gross histological work.
Graduate Laboratory, Missouri Botanical Garden.
* Durand, E. J, The differential staining of intercellular mycelium. Phyto-
pathology i: 129-30. 1911.
1
TWO TRUNK DISEASES OF THE MESQUITE
HERMANN YON SCHRENK
Pathologist to the Missouri Botanical Garden
The diseases of the mesquite (Prosopis glandulosa Torr.)
hitherto recorded are comparatively few in number; Heald and
Wolf (5) enumerate seven from southern Texas as due to fungi.
The pods are frequently affected by an anthracnose, GloBosporium
leguminum (Cke,) Sacc; the leaves are attacked by Cercospora
proiopidis Heald and Wolf, a species of powdery mildew {Ery-
siphe ?), and by a rust, Ravenelia arizonica Ell. & Ev. ; and a
leaf blight due to some unknown cause is also mentioned. The
large limbs and smaller branches show galls, evidently not due
to insect attack, and the mistletoe {Phoradendron flavescens
(Pursh) Nutt. is sometimes destructive- In addition to the
above, the writer has frequently noted the weakening effect,
particularly near the ends of branches, brought about by vigor-
ous growths of the ball moss {Tillandsia recurvata L.). Birge
(1) has given a good description of the effects of this plant on
trees in Texas.
Of the insect injuries of the mesquite, that of the mesquite
hoTi)r (Cyllene antennatus White) is of interest. The insect is
described by Horn (6) as attacking mesquite wood in Arizona,
but no description of its work is given. While T have not seen
the insect at work in Texas, the holes found in the mesquite
trees are so like those described for other species of Cyllene^
notably Cyllene robinice Forster (10) — which attacks the locust
— that the assumption seems w^arranted that the Texas insect
is the one referred to by Horn. The tunnels extend straight
■
through the bark into the heart-wood, and up and down in the
latter, thus forming ideal channels for the entrance of fungous
spores.
The only reference to trunk diseases which has been found is
a brief statement by Havard (4), in an account of the mesquite,
in which he mentions that '' unfortunately it too often happens
that the zones of the heart-wood are fissured, decayed or de-
Ann. Mo. Bot. Gaed., Vol. 1, 1914
(243)
244 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
taclied from each other, so that it is difficult to get flawless
boards."
In 1912 the writer found the older mesquite trees in the
vicinity of San Antonio, Texas, seriously affected by a trunk
disease, caused by one of the polyporous fungi. In one small
field some twenty or more trees were found bearing the fruiting
bodies of this fungus. Its distribution in the vicinity of San
Antonio was general, and it is probable that it extends over a
wider range, as evidenced by the finding of a sporophore by
Underwood in the vicinity of Austin, in 1891.
Where the mesquite develops into a bush with several trunks,
sometimes only one of the several trunks is affected, but in
other cases several or all of them contract the disease. The
age of the affected trees was difficult to estimate. The mesquite
grows rather rapidly at first, but very slowly after eight or ten
years. According to Sargent, trunks thirty years old may be
seven to eight inches in diameter, while trees one foot in diam-
eter are probably over one hundred years old. The trees
found affected were from two to ten inches in diameter and
all over twenty years of age, some of them probably very much
older.
The decay is confined entirely to the heart-wood of the main
trunks, extending from the ground up into the trunk for varying
distances. The distribution is such that it is obvious that the
fungus gains entrance through wounds in the trunk above the
ground, chiefly through old branch stubs and borer holes, as is
so frequently the case with trunk diseases of this kind. One
instance was found which made it appear obvious that the holes
made by the borer had served to give the fungus a start.
Sections of diseased trunks showed that the heart-wood was
decayed to a greater or less degree (pi. 6 fig. 2). Mesquite
wood has very sharply defined heart and sap-wood. The latter
is light yellow or almost white and very narrow, being composed
of but a few rings of wood, whereas the heart-wood is rich brown
or reddish. The decay of the heart-wood begins near the center,
and gradually spreads outward towards the bark; there is very
little, if any, change in color (except that the decayed wood is a
lighter shade of brown), and here and there irregular, thin lines
of undecayed wood can be seen extending through the diseased
1914)
VON SCHRENK — TWO TRUNK DISEASES OF MESQUITB 245
part. The decayed wood is very brittle, but still remains fibrous,
that is, it does not crumble into powder like charcoal. It splits
like sound wood, but is spongy and soft. The wood of the
mesquite is very hard and heavy, a cubic foot weighing 47.69
pounds when absolutely dry. It consists of numerous, distinct
medullary rays, and distinct but irregularly distributed bands
of very thick-walled wood fibers, between which occurs a
thinner-celled wood parenchyma. In the heart-wood the
lumina of the cells of the latter tissue are usually completely
filled with a yellow-brown substance, largely composed of tan-
nin. McMurtree (8) found tannic acid in large quantities in
mesquite wood, 6.21 percent in the heart-wood, 0.5 per cent in
the sap-wood, and 0.5 per cent in the bark. Besides tannin
he found of materials other than tannin, insoluble in water but
extracted by ether, 0.6 per cent in the heart-wood, 6.7 per cent in
the sap-wood, and 1.84 per cent in the bark. A considerable
number of large, open ducts are found in the early part of each
wood ring. These also are filled with a yellow-brown substance
similar to that found in the wood parenchyma.
The fine, colorless mycelium of the fungus spreads throughout
the wood substance. Unlike Polyporus rimosus in locust wood
(10), the fungus does not destroy the wood as a whole, but
attacks only the heavily lignified groups of wood fibers. These
are wholly destroyed, leaving holes or gaps between the vessels
and wood parenchyma. The dissolution of the wood fibers evi-
dently proceeds with great rapidity, starting with the secondary
thickening of each cell. The cells disappear entirely, and in
advanced stages of decay small masses of mycelium are the only
evidence of their former presence. Although the wood paren-
chyma and the vessels are filled with hyphae, they resist destruc-
tion almost completely,^a fact which may be connected with
the very high tannin content of both of these tissues. The
recent results of Wehmer (11), who found that for certain
species of fungi tannin exerts a retarding influence on develop-
ment, and the similar findings of Knudson (7), and of Cook and
Taubenhaus (3), who state that "tannin has a tendency to
retard or inhibit the growth of fungi," and that "the parasitic
forms are more sensitive to the action of tannin than the sapro-
phytic forms," lend support to this idea. Cook and Taubenhaus
[Vol. 1
246 ANNALS OF THE MISSOURI BOTANICAL GARDEN
also found that for the parasitic fungi tested, concentrations of
from 0.1 per cent to 0.6 per cent were sufficient to retard growth.
While the mere presence of considerable tannin may not entirely
prevent the development of a fungus, it may retard its growth,
and in the mesquite may explain the comparative immunity of
the wood parenchyma to its attacks. The selective destruction
of the wood fibers will serve to distinguish this form of decay
from the other types of hardwood decay.
From the material found it was not possible to judge of the
ultimate stages of the disease. In view of the fact, however,
that sporophores four years old were observed, it seems that the
resistance of a part of the wood structure is more or less per-
manent. No mesquite trees were found broken off as a result of
the action of the fungus. It is conceivable, however, that very
severe storms might break off trees weakened by the disease. '
^ The fungus which causes the decay is Polyporus texanus (Mur-
rill) Sacc. & Trott. The sporophores, which are annual and
very distinct and easily recognized, develop around old knots.
At the end of one year the sporophore dries and cracks (pi. 6
fig. 1, and pi. 7 figs. 1,2), and many of them become badly eaten
by insects. The latter may completely destroy the fruiting
structure, thereby preventing the formation of new pilei from
the original one. The sporophores occur either singly or in
groups. In the latter case the oldest sporophore of the group
is situated near the trunk, and gives rise during the second year
to another pileus; from the latter a third one may grow out
during the following year. This habit is well shown in pi. 6 fig.
1, and in pi. 7 fig. 1. The photograph reproduced in pi. 6 fig. 1
shows a group of three sporophores from below; the oldest one
(in the back), dried and cracked; the second one formed immedi-
ately below the oldest one; and the youngest one developed at
the side. This condition is also evident in pi. 7 figs. 1, 2. On
the trees observed there was usually only one sporophore or a
single group of sporophores, and while the internal decay ex-
tended in some cases for ten to twelve feet up and down in the
trunk, in no case did the sporophores develop at more than one
point.
Polyporus texanus (Murrill) Sacc. & Trott., was first described
by Murrill (9) in 1904 from a specimen collected by Under-
1914]
VON SCHRENK — TWO TRUNK DISEASES OF MESQUITE 247
wood on a mesquite (?) tree near Austin, Texas, in 1891. Mur-
rill's description of this fungus is as follows :
"Pileus ungulate, attached by the vertex, 3x5x4 cm., surface
rimose
age, the separated areas imbricated; margin very obtuse, concolor-
ous, context corky, concentrically banded, fulvous to umbrinous,
very thin, only one-tenth the length of the tubes in thickness; tubes
mm
guttulate
^Tiile this description was made from one specimen, the
characterization is a good one and well defines the sporophores
recently collected, and now in the herbarium of the Missouri
Botanical Garden. One of the marked characters of the fruit-
ing structure is the concentrically and radially rimose surface
(pi. 7 figs. 1, 2) with imbricated areas, particularly in the older
specimens. The tubes are very long, 2-3^ cm. (as stated by
Murrill), and make up the larger part of the mass of the sporo-
phore. The largest specimen found measured 9.5 cm. in width,
7 cm. in length, and 5 cm. in thickness. Using Ridgeway's
color scale, the top is avellaneous gray, the tubes tawny, the
substance antique brown (umbrinus of Saccardo's scale); near
the margin the color is verona brown to warm sepia. Murrill's
statement that the sporophore is attached by the vertex should
be amplified, as many of the sporophores are practically dimidi-
ate. With the additional material now available for study, the
modified description of the fungus in question is as follows :
L
Polyporus texanus (Murrill) Sacc. & Trott. Syll. Fung. 2i:
272. 1912.
Inonotus texanus Murrill, Bull. Torr. Bot. Club 31 : 597. 1905.
Pileus ungulate, attached by the vertex or dimidiate, 4-9.5
cm. wide, 3-7 cm. long, and 4-5 cm. thick; surface avellaneous
gray to fulvous, concentrically and radially rimose, especially
in age, the separated areas imbricated; margin very obtuse,
verona brown to warm sepia; context corky, concentrically
banded, antique brown, very thin, only one-tenth the length of
the tubes in thickness; tubes 2-3^ cm. long, 2-3 to a mm., tawny,
polygonal, edges thin, entire; spores ovoid, smooth, very dark
brown, 1-2 guttulate, 8 x 10 /z. Parasitic on living mesquite trees.
-^40 ANNALS OF THE MISSOUKI BOTANICAL GARDEN
In the same locality in which Pohjporus iexanus occurred, one
mesquite tree was found bearing a sporophore of Fomes rimosus
Berk. This fungus causes the heart rot of RoUnia Pseudo-Acacia
(10), and it is of interest to note its occurrence on a new host.
The specimen found is a typical sporophore of Fomes rimosus,
measuring about two inches in length; unfortunately it was not
recognized at the time of collection, and sections of the affected
tree were therefore not made. In view of the destructive char-
acter of this fungus when found on Rohinia, however, it is prob-
able that it causes a similar heart rot of the mesquite. Further
search will be made in the San Antonio region for additional
evidences of its occurrence.
The wood of the mesquite is usually described as being very
resistant to decay after it has been cut from the tree. For many
years mesquite posts have been used in the southwest in prefer-
ence to other kinc^s. Mesquite ties, foundation posts, etc.,
have also proved that the wood is very resistant to decky. This
applies only to the heart-wood, however. The sap-wood is very
short-lived, and where small trunks are cut, as is now frequently
the case, and used for fence posts, the length of life is very short,
ometimes not over two to three years. The destruction of
the sap-wood is due to a number of insects and saprophytic
fungi, all of which are common on dead branches, posts, etc.,
in the vicinity of San Antonio. Of the more common fungi, the
following were recently collected: Polystictus Lindheimeri B. <?.
C., Stereum Leveillianum Fr., Schizophyllwn commune Fr., Len-
zites proiractus Fr., and Stereum alhohadlum Schw.
The author acknowledges assistance from the following:
Mr. Kearney Mason, of San Antonio, for permission to fell
trees on his land and assistance in doing so; Dr. E. A. Burt,
Mycologist and Librarian to the Missouri Botanical Garden,'
and Mr. C. G. Lloyd for aid in the identification of species of
fungi.
1914]
VON SCHRENK — TWO TRUNK DISEASES OF THE MESQUITB 249
Literature Cited
1. Birge, W. L. The anatomy and some biological aspects of the "Ball Moss/*
TiUandsia recurvata L. University of Texas Bui. 194: 1-24. 1911.
2. Bray, Wm. L. The mistletoe pest in the Southwest. U. S. Dept. Agr., Bur.
PL Ind. Bui. 166: 1-39. 1910. [cf. pp. 18, 25.]
3. Cook, M. C, and Taubenhaus, J. J. The relation of parasitic fungi to the con-
tents of the cells of the host. Part I. The toxicity of tannin. Del. Ag. Exp.
Sta. Bui. 91: 1-67. 1911.
4. Havard, V. The Mezquit. Am. Nat. i8: 451-59. 1884. [cf. p. 457.]
5. Heald, F. D., and Wolf, F. A. A plant disease survey in the vicinity of San An-
tonio, Texas. U.S. Dept. Agr., Bur. PL Ind. Bui. 226: 1-112. 1912. [cf.p. 72.)
6. Horn, G. H. American Coleoptera. Trans. Am. Entom. Soc. 8: 135. 1880.
7. Knudson, L. Tannic acid fermentation. Jour. Biol. Chem. 14:159-202, 1913,
8. McMurtree, Wm. Rept. U. S. Commissioner of Ag. for 1875 182. 1876.
9. Murrill, Wm. The Pol3Tporaceae of North America — IX. Bull. Torr. Bot. Club
31: 593-610. 1904. [cf. p. 597.]
10. von Schrenk, Hermann. A disease of the black locust. Ann. Rept. Mo. Bot.
Card. 12: 21-31. 190L
11. Wehmer, C. Der wachstumshemmende Einfluss von GerbsSuren auf Merulius
lachrymans in seiner Beziehung zur Resistenz des Eichenholzes gegen Haus-
schwamm. Mycologisches Centralbl. i: 138-48. 1912,
4
9
[Vol. 1, 1914]
250 ANNALS OF THE MISSOURI BOTANICAL GARDEN
i
1
Explanation of Plate
PLATE 6
Disease of the mesquite due to Polyporus texanus
FiQ. 1. View showing the manner in which a group of sporophores of Polyporus
texanus grows on the trunk; also the lower surfaces of the sporophores.
Fig. 2. Two sections of diseased mesquite trunk showing the manner in which
the wood is destroyed.
4
Ans. Mo. Bot. Gaud., Vol. I. 1914
Plate 6
Pig. 1.
FIG. 2.
VON SCHRENK — TRUNK DISEASES OF MESQUITE
^ .
\
\
^
\
COCK A Y \ K. HOSTi ) N.
[Vol. 1, 1914
252 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 7
Disea3e of the mesquite due to Polyporus texaniis
Fig. 1. Side view of a group of sporophores of Polypoms texamis growing on a
living mesquite tree.
FiQ, 2. Front view of a group of aporophorea of Polyporus texanus growing on a
living mesquite tree.
Ann. Mo. Bot. Gard, Vol. 1. 1914
Plate 7
FIG. 1
Fig. 2.
VOX SCHRENK — TRUNK DlSEAvSES OF MESQUITE
COC K A Y N E . BOSTO N .
I
'J
A TRUNK DISEASE OF THE LILAC
1 HERMANN VON SCHRENK
Pathologist to the Missouri Botanical Garden
A general discussion of diseases of the common lilac {Syringa
mlgaris L.) was recently published by Klebalm (3). This
author enumerates a number of diseases, such as the one of
bacterial origin ascribed to Pseudomonas Syringm, various leaf
: diseases due to species of Microsphoera, Gloeosporium, and other
» leaf parasites, and a disease due to Botrytis cinerea. The
major part of the work, however, deals with a disease due to
Heterosporium Syringce Oud., affecting the leaves, and a serious
twig blight due to Phytophthora Syringce Klebahn. Subsequent
papers by various writers deal with one or the other of the
diseases mentioned by Klebahn.
I During recent years a destructive trunk disease of the com-
mon Ulac (Syringa mlgaris L.) has been noted a number of
timers in the Missouri Botanical Garden, and in grounds in the
vicinity of St. Louis. The affected plants were usually old
bushes which had been more or less neglected, and the tops of
the leading trunks were frequently dead. Long shoots from the
root and others from the part of the trunks near the ground made
a dense tangle around the main stem; on the latter sporophores
of Polyporus versicolor were found in various stages of develop-
ment, sometimes isolated, but more frequently in groups. Sec-
tions were made of the trunks on which this fungus was growing
and it was found that such trunks were invariably diseased,
while those close by, either from the same root system or from
adjacent bushes — which were free from the fungus — were al-
ways sound.
In pi. 8 fig. 1 two affected trunks are shown cut at points
about three feet from the ground. In both cases the larger
part of the stem was aUve, as evidenced by the presence of
vigorous shoots along the entire length. PI. 8 fig. 2, and pi.
9 figs. 1, 2 represent sections of lilac trunks taken from different
bushes to show different stages of the disease.
T
T
(
Ann
7
(253)
254 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
The wood of the lilac is white in color, hard, and close-grained.
In younger trunks there is no appreciable difference between
heart-wood and sap-wood; as the trunks grow older, however,
the heart-wood turns darker, and in those twelve years old, or
thereabouts, it is distinctly darker than the rather thin, white sap-
wood.
The disease first manifests itself in the inner heart-wood,
frequently in close proximity to the holes made by the lilac
borer. This lepidopterous insect {Podosesia syringce Harris)
(for whose identification I am indebted to Dr. E. P. Felt) has
been found very destructive to lilac bushes, and, according to
Beutenmiiller (1), occurs from New England and the middle
states westward to Colorado and southwest to Texas. Quoting
from Beutenmtiller's account: ''The female deposits her eggs
in patches on roughened or knotty places on the bark of ash and
lilac. The eggs, according to Hulst, hatch in about six days,
and the newly born larvae at once eat their way through the
bark into the solid wood. They run their channels longitudi-
nally for about 8-10 inches through the wood. The larvae
pupate in slight cocoons after cutting their way to the bark, of
which they leave only a thin outer skin. The pupation usually
takes place early in May, and the moths emerge in about three
weeks." Felt (2) briefly described the habits of the larva,
stating that "a sign of its presence in midsummer being largely
the sudden wilting of a shoot. " He quotes from an observation
made by Dr. Kellicott in which the latter states that he ''watched
20 or more issue from a single tree in one day, and found that
often there were more than one hundred in one tree." Felt
recommends cutting and burning all infested wood in the early
spring.
In the vicinity of St. Louis the lilac borer has been very active
in recent years, judging from the fact that very few lilac bushes
over five years old were found free from its attacks. Without
much doubt the fungous spores get into the interior of the lilac
trunks through the borer holes, and start to develop within the
heart-wood on the edges of the borer holes. In pi. 9 fig. 1 two
borer holes, still filled with pieces of the borings, can be seen in
the lower right-hand trunk, and one small hole in this same sec-
tion occurs in the sap-wood. The fungus, after it has begun to
1914]
VON SCHRENK — A TRUNK DISEASE OF LILAC 255
grow in the hole, rapidly spreads up and down in the heart-
wood, and soon grows out from the center toward the bark. As
the disease progresses, the wood is converted into a soft, pithy,
white mass, having the consistency of corn-stalk pith. The
line of demarcation between the sound and completely destroyed
wood is very sharp (see pi. 9), resembling in this respect the
type of decay caused by this same fungus in living catalpa trees
((5), pi. 26). The line between sound and decayed wood is so
sharp that entirely decayed fibers adjoin perfectly sound ones.
Between the wholly unaffected wood and the completely de-
stroyed fibers, is a narrow ring of darker wood, which is, to all
intents and purposes, sound ; the wood cells are partially invaded
by the mycelium of the fungus, and the lumina are filled with a
yellow-brown liquid, which when seen in mass gives the section
the dark color referred to. This liquid dries out in some places
and leaves a brown amorphous substance, such as has frequently
been found in the early stages of decomposition of hardwood
wood fibers (6). It probably consists of decomposition products
which are infiltrated into the sound wood immediately in ad-
vance of the fungus. In cases where the fungus starts in several
centers, rings of the darker colored wood surround each decayed
portion, a condition which is well shown in pi. 9 figs. 1, 2, where
the fungus is growing in the center of the trunk and in addition
in three more peripheral localities. In the lilac the brown sub-
stance referred to is ultimately destroyed (see the middle trunk
of the lower tier, pi. 9 fig. 2, where the wood is destroyed up to
the bark).
The completely decayed wood, which readily absorbs water,
resembles pith, and in general is very similar to catalpa wood
destroyed by Polyporus versicolor (5)- It has some of the
attributes of wood, i.e., it can be split, is fairly compact, and
cannot be crumbled into powder. Sound lilac wood is very
heavy and hard, and is composed almost wholly of very thick-
walled wood cells, with small vessels scattered with considerable
regularity throughout the annual ring; wood parenchyma is
almost wholly absent. The hyphse of Polyporus versicolor
attack and very rapidly destroy the layers of secondary thick-
ening of the wood cells. The middle laniellce retain the nature
of lignified fibers and resist destruction alnost entirely, although
[Vol. 1
256 ANNALS OF THE MISSOURI BOTANICAL GARDEN
here and there some of them are dissolved, giving rise to small
separated cell groups. Entire dissolution rarely takes place
(this was also found to be true for diseased catalpa wood ((5)
pi. 52)). With the removal of the secondary thickening, the
resulting decayed wood has a skeletonized appearance. It
has all of the elements, but these are very thin-walled. The
fine medullary-ray cells are destroyed here and there, producing
radial, isolated masses, but more frequently the decayed mass
hangs together firmly. The dissolution of the layers of secon-
dary thickening goes forward very evenly, bringing about the
sharp dividing line between sound and decayed wood already
referred to.
The only difference between the catalpa and lilac diseases is
that in the catalpa the entire wood mass is skeletonized, whereas
in the lilac hard areas of undestroyed wood fibers are left here
and there, surrounded by decayed wood (pi. 9 figs. 1,2). These
masses are either entire rings (pi. 9 fig. 1) or irregular areas
lying detached within the decayed parts, and represent portions
of the heart-wood which for some reason have temporarily es-
caped total destruction; the wood fibers are filled with the yellow-
brown substance, but do not otherwise differ from normal wood
fibers. As the disease progresses, however, they are finally
destroyed. This was made evident by the fact that in the
upper parts of diseased trunks these immune areas were always
found coexistent with the early stages of the disease, while
lower down in the trunks, where the advanced stages of decay
had been reached, they were practically absent. The temporary
immunity may be due to the presence of more resistant groups
of wood fibers, possibly also to a high concentration of decom-
position products.
The development of the fungous mycelium from the center of
the trunk out toward the bark differs radically from that of any
other disease known to the writer. In most trees the destruc-
tion of wood by a fungus growing in the dead heart-wood is
confined to the latter, — further growth ceasing as soon as the
mycelium reaches the sap ring. As has been suggested by
Munch (4), this is probably due to the fact that most mycelia
of wood-destroying fungi require a balance between the amounts
of oxygen and water contained in the wood fiber. Any undue
1914]
VON SCHRENK — A TRUNK DISEASE OF LILAC 257
percentages of either may make the conditions unfavorable for
further development.
In the lilac disease the fungus may grow outward concentrically
in a regular manner (pi. 9 fig. 1). Very frequently, however,
the fungus grows out into the sap ring at one side, at first slowly,
then more rapidly. This is well shown in pi. 9 fig. 2, where
four successive stages are represented by photographs. In
the upper left-hand trunk the fungus has almost reached the
barkj and in the three lower ones it has reached the bark and is
gradually killing it. The probable explanation for this behavior
is to be sought in the water content of the wood fibers. It was
found that in many cases where the fungus actually grew up to
the bark and through it, that on that side the lilac borer had
been active; the wood fibers in the vicinity of the holes dried
sufficiently to make growth possible for the mycelium, and as
the destruction took place more drying occurred in adjacent
areas until ultimately the whole sap region on that side was
invaded and destroyed.
A number of water determinations were made of the wood
fiber in the immediate vicinity of the growing mycelium, and
the results compared with those obtained from normal sap-wood.
In all cases the sap-wood about to be invaded was found to
have a very much lower water content than the normal sap-
wood. Unfortunately, it was impossible to get exact data which
would indicate accurately the highest moisture content at which
growth was possible; infected wood had obviously already
reached and gone beyond that point, and as to sound new wood,
even that which was near the borer holes, nothing could be
postulated with certainty concerning its susceptibility or non-
susceptibility to fungous attack. It would be an interesting
problem to test the water susceptibility of Polyporus versicolor
in its relation to lilac wood. It seems probable, however, that
the drying out of one side of the trunks was at least one of the
determining factors in the rather striking and exceptional
method of growth of the fungus. Whether the fungus would
eventually have destroyed the entire trunk it is impossible to
state, because no such wholly destroyed trunks were found.
There seems to be no reason, however, why this should not
258 ANNALS OF THE MISSOrRl BOTANICAL GARDEN
rvci.. 1
occur; in fact, the right-hand trunk of the lower tier in pL 9
fig. 2 has very little live wood left.
After the mycelium has reached the bark it grows through it,
and fruiting bodies develop on the outside. The latter some-
times occur singly, but more commonly in linear groups parallel
to the long axis of the trunk. Frequently one or more fruiting
bodies grow out from the holes made by the borer. In the
right-hand trunk in pi. 8 fig. 1 sporophores are shown growing
out at the base of vigorous, live shoots; in the left-hand trunk
the shoot is dead, having been killed during the year as the
fungus invaded the wood from which the shoot was growing.
The sporophores found were typical of Polyporus versicolor L.
This fungus is so common on the dead wood of various hard-
woods that a detailed description is hardly necessary. It is
interesting to note here that this is the second instance where
this fungus attacks living plants. In the case of the catalpa the
fungus grows vigorously only in the live tree; infected wood
rarely, if ever, is decayed after it is cut from the tree. Many
thousand posts of catalpa, the heart of which had been partially
destroyed by Polyporus versicolor, have served as fence posts
during the last ten years without showing a sign of decay of that
part of the wood which was sound at the time of cutting. With
the lilac it is different; the dead wood is just as subject to attack
as is dead oak, beech, or gum wood.
The age at which lilac bushes are attacked has not been defi-
nitely determined. Those examined were about 15-20 years old
(2^ inches in diameter) . The trunk shown in pi. 8 fig. 2 was over
thirty years old. It is probable that the disease is not serious
until the bushes are ten or more years old, although this will
depend somewhat on the rate of growth. Trunks l|-2 inches
in diameter were frequently found diseased. The effect of the
disease is to gradually kill the top of the trunk; side shoots then
develop farther down, which in turn are killed by the fungus,
and eventually the trunk is broken off by the wind or snow.
The prevention of the disease is possible by continued atten-
tion to the borers. A careful examination for the latter should
be made in June or July, and if any are present these should be
killed by means of a wire, and the holes— after antiseptic
treatment with some coal-tar compound — plugged. Painting
»
1914]
VON SCHRENK — A TRUNK DISEASE OF LILAC 259
m
or brushing the lower parts of trunks with whale oil soap, or its
equivalent, should also prove of value. Wherever a diseased
trunk is found it should at once be cut out and burned. All
dead wood in the neighborhood of lilac bushes should be cleaned
up, so that the chances for infection may be reduced.
Literature Cited
1. Beutenmliller, Wm. Monograph of the Sesiida. Mem. Am. Mub. Nat. Hist.
i:244. 1903.
2. Felt, E. P. Insects affecting park and woodland trees. Rep. N. Y. State Mus.
59»: 8-332. 1905. [of. p. 104.]
[For other insects attacking lilac see Rep. N. Y, State Mus. 59*: 839. 1905.]
9. Klebahn, H. Krankheiten des Flieders 1-60. Berlin. 1909. llllust.J
4. MQnch, Ernst. TJntersuchungen Uber Immunitat und KrankheitsempfSnglichkeit
der Holzpflanzen. Naturwiss. Zeitschr. f. Forst u. Landw. 7:54-75,
87-114, 129-60. 1909.
-6. von Schrenk, Hermann. The diseases of the hardy catalpa. U. S. Dept. Agr.,
Bur. Forestry Bui. 37; 51-58. 1902
e. , A disease of white ash. U. S. Dept. Agr., Bur. PL Ind. Bui. 32: 1-20.
1903. [cf. p. 14.1
260 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1, 191 4J
• .
Explanation of Plate
PLATE 8
Trunk disease of lilac due to Polypoms versicolor
Pia. 1. View of two diseased lilac trunks showing the sporophores of Polyporu^
versicolor^ and the manner in which living branches grow from diseased trunks.
Fia. 2, Sections of an old diseased lilac trunk showing the decayed heart-wood
Ann. iMo, Bur. Ctard. Vol. 1. 1934
Plate R
\-
\
\
Fig, I
mm. 3
Fic. 2
VOX SCHRENK — TRUNK DISEASE OF EIEAC
COC KAY N'K. IIOSTON.
262
[Vol. 1, 1914]
ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 9
Trunk disease of the lilac due to Polj/porus versicolor
Fig. 1. Sections of three diseased trunks showing early stages of the disease
Fia. 2. Sections showing progressive stages of the lilac trunk disease.
Anm. Mo. Box. Gakd,Voi.. 1. 1914
J'late y
Fig. 1
FIG. 2.
VON SCIIR.ENK — TRUNK DLSKASR OF LILAC
COCKAYXE. rOSTON.
F
Annals
of the
Missouri Botanical Garden
Vol. I SEPTEMBER, 1914 No. 3
DESCRIPTIONS OF NORTH AMERICAN SENECIONE^^
J. M. GREENMAN
Curator of the Herbarium of the Missouri Botanical Garden
Associate Professor in the Henry Shaw School of Botany of
Washington University
The following descriptions and notes are the results obtained
from a critical study of material in several herbaria during the
preparation of a monograph 'of the North American species of
the genus Senecio. Some of the species here described have
been in manuscript a number of years and a few of them have
been withheld from pubHcation, because of incomplete speci-
mens, hoping that additional material might be brought together
before pubhcation. In many cases supplementary and sub-
stantiating material has been obtained from which it is now
possible to make fairly complete diagnoses. In one or two
instances a reconsideration of certain natural groups within the
genus, in the light of recent collections, has made it possible to
combine forms which formerly were taken to represent distinct
species. Very few new species have resulted from recent col-
lections, but there are still many regions, particularly in Central
America, which are inadequately explored. The writer would
welcome material in this genus from any part of North America
in order that the geographical range of species may be recorded
as ac(;urately as possible in his forthcoming monograph. The
sections indicated in parentheses immediately following the
generic name are in accordance with my preliminary paper to
which reference is made under the snecies.
Issued September 30, 1914.
Ann. Mo. Bot. Gabd., Vol. l, 1914
(2G3)
[Vol. 1
264 ANNALS OF THE MISSOURI BOTANICAL GARDEN
4
Senecio (§ Aurei) hyperborealis Greenm. Monogr. Senecio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
S.resedijolius Hook.Fl.Bor. Am.i : 333. pi 117. 1833, not Less.
Herbaceus perennis; caule simplice vel ramoso suberecto 1-2
dm. alto plus minusve foliaceo juventate glabro vel parce floc-
culoso-tomentuloso S2epe ad basin et in axillis foliorum per-
sistenter lanato-tomcntoso; foliis inferioribus petiolatis indivisis
vel plerumque irregulariter lyrato-pinnatifidis 4-10 cm. longis
1-2.5 cm. latis, lobis remotis; foliis superioribus multum reductis
sessilibus et bracteiformibus; capitulis paucis terminalibus radi-
atis 10-12 mm. altis 2-3.5 cm. (radii inclusis) diamctro; floribus
femineis 10-12, ligulis flavis 10-12 mm. longis ca. 2 mm. latis;
disci flosculis numerosis; achseniis sa^pe paulo hispidulis.
Specimen examined:
Canada: Arctic America, Hooker (Gray Herb.), type.
Var. columbiensis (Gray) Greenm. Monogr. Senecio, pt. 1,
24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
/S. resedif alius var. columbiensis Gray, Syn. Fl. i^: 390. 1884.
Habitu formae typicse; capitulis heterogamis, ligulis florls
femineis quam squamis involucri paulo brevioribus; acha^niis
glabris.
Specimen examined :
British Columbia: Mucklung River, 25 July, 1882, Mr. Mackay
(Gray Herb.).
Senecio (§ Lobati) prolixus, comb. nov.
S. diffusus Greenm. Monogr. Senecio, pt. 1, 24. 1901; in
Engl. Bot. Jahrb. 32: 20. 1902, nomen, not Linn. f.
Herbaceus perennis glabrus vel in axillis foliorum albo-
tomentosus; caule tereti striato simphci vel ramoso erecto 2-5
dm. alto; foliis petiolatis vel sessilibus inferioribus lyrato-pin-
natifidis petiolo incluso usque ad 15 cm. longis 1.5 5 cm. latis
utrinque glabris, segmentis lateralibus oblongo-cuneatis cum
sinis altis rotundatis disjunctis granditer dentatis, superioribus
remotis sessilibus pinnatifidis sursum multum reductis; inflores-
centiis laxe corymboso-cymosis 1-2.5 dm. diametro; capituhs
circiter 1 cm. altis radiatis; involucris campanulatis parce cal-
yculatis glabris; involucri squamis plerumque 21 lanceolatis
vel lineari-lanceolatis 5-6 mm. longis acuminatis acutis; flos-
culis liguliferis ca. 13, ligulis oblongis 5-6 mm. longis flavis;
1914]
GREENMAN— NORTH AMERICAN SENECIQNE^ 265
^ ri
floi-ibus disci numerosis 50-60; achteniis maturitate 2-3 mm.
longis striatis glabris.
Specimens examined:
California (?): "Mohave Region," April-May, coll. of 1884,
J. G. Lemmon, 3130 (Gray Herb.), type.
Arizona: Wickenburg, valley of the Hassayampa River, April,
1876; Dr. Edward Palmer, 6I4 (Gray Herb, and Mo. Bot.
Gard. Herb.).
The specimens cited may be looked for in herbaria under
S. muUilobatus Torr. & Gray, to which the species here proposed
is related, but from which it differs in well developed specimens
in the outline and size of the leaves, loose inflorescence, and
larger heads with 21 instead of 13 involucral bracts. S. pro-
lixus has rather more the aspect of S. Breweri Davy.
Senecio (§ Tomentosi) appendiculatus Greenm. Monogr.
Senecio, pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32 : 20. 1902, nomen.
S. neo-mexicanus Gray, Proc. Am. Acad. 19: 55. 1883, in part;
Syn. Fl. i^: 392. 1884, in part, as to plant of Thurber.
Herbaceus perennis ubique plus minusve albo-tomentosus;
caulibus subcsespitosis erectis 1.5-3 dm. altis striatis ssepe foli-
aceis; foliis radicalibus oblanceolatis vel oblongo-obovatis
petiolo incluso 3.5-10 cm. longis 0.5-2 cm. latis dentatis ad
basin in petiolum paulatim angustatis integris, eis caulinis petio-
latis vel sessilibus 2-7 cm. longis ad basin plerumque ampliatis
irregulariter dentatis subamplexicauhbusque; inflorescentiis ter-
minalibus corymboso-cymosis 6-12-cephahs; capitulis 10-12
mm. altis radiatis; involucris campanulatis minute calyculatis;
involucri squamis plerumque 21 lanccolatis 5-7 mm. longis
acutis sparsissime tomentulosis; flosculis liguHferis ca. 13, HguHs
flavis; fioribus disci numerosis ca. 70; achaeniis glabris.
Specimens examined:
New Mexico: Mule Spring, May, 1851, Geo. Thurber, 280 (Gray
Herb.), type; Organ Mountains, Dona Ana Co., 25 April,
1907, E. 0. Wooton, 3370 (Mo. Bot. Gard. Herb.).
This species is related to S. neo-mexicanus Gray, to which it
has been usually referred, but from which it differs in having
a more leafy stem, undivided leaves, and with the stem-leaves
commonly ampHated into a more or less dentate half-clasping
base, and finally in having glabrous instead of hirtellous achenes.
[Vol. 1
266 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Senecio (§ Tomentosi) convallium Greenm. Monogr. Senecio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
Herbaceus perennis ubique sericeo-pubescentes; caulibus
csespitosis erectis 3 dm. altis; foliis inferioribus rosulatis petiolatis
elliptico-lanceolatis vel oblongo-oblanceolatis 2.5-6 cm. longis
5-12 mm. latis acutis integris vel supra mediam partem pauci-
dentatis basi longe cuneatis integriusculis juventate utrinque
sericeo-pubescentibus setate supra plus minusve glabratis, foliis
superioribus spathulato-oblanceolatis angusti-petiolatis; inflores-
centiis corymboso-cymosis paucicapitatis; capitulis circiter 1
cm. altis subradiatis; involucri bractcis 13-15 lineari-attenuatis
7-9 mm. longis acutis sparse sericeo-tomentulosis; floribus
femineis subligulatis; floribus disci 30-35; achaeniis 3.5 mm.
longis striatis glabris.
Specimen examined :
Utah: Rabbit Valley, altitude 2130 m., August, 1875, L. F.
Ward, 704 of the "U. S. Geological and Geographical Sur-
vey of the Territories" (Gray Herb.), type.
The species here characterized has been hitherto confused
with S. canus Hook., from which it is readily distinguished by
the subsericeous pubescence and technical characters of the
head.
Senecio (§ Tomentosi) kernensis Greenm. Monogr. Senecio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
Herbaceus perennis ubique dense lanato-tomentosus; caule
tereti erecto ca. 1 dm. alto; fohis inferioribus rosulatis petiolatis
elliptico-oblongis vel oblongo-rotundatis 1-3 cm. longis 3-10
mm. latis apice obtusis vel rotundatis basi abrupte angustatis
vel subtruncatis utrinque dense lanato-tomentosis, marginibus
integris vel subcrenato-dcntatis revolutisque, foliis superioribus
bracteiformibus multum reductis; inflorescentiis terminalibus co-
'>-pvmnsis nnnpinnnitfl.tis! nanitulis 8 10 mm. altis radiatis
rymboso-cymosis paucicapitatis; capitulis 8 10
5-8 mm. (radii cxclusis) diametro parce calyculatis; involucri
squamis ca. 13 lineari-lanceolatis 5-0 mm. longis acutis floccoso-
tomentulosis subglabratis; achaeniis glabris.
Specimen examined:
California: South Fork of Kern River, altitude 3760 m., Septem-
ber, 1875, Dr. J. T. Rothrock, S34 of the ''Explorations and
Survevs west of the 100th Meridian" (Gray Herb.), type.
ca.
GREENMAN — NORTH AMERICAN SENECIONE.E Zb/
Senecio (§ Tomentosi) macropus Greenm. Monogr. Senecio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
S. arizonicus Gray, Syn. Fl. i^: 392. 1884, in part, as to plant
of Rusby.
Radix robusta in sicco 2.5 cm. diametro; caulibus erectis usque
ad 7.5 dm. altis glabris vel in axillis foliorum albo-tomentulosis
striatis plus minusve purpurascentibus; foliis radicalibus petio-
latis lyrato-pinnatifidis petiolo incluso 10-14 cm. longis 4-5 cm.
latis, segmentis paucijugis insequalibus terminali njajore ovato-
oblongis 5-0 cm. longis grosse dentatis, ceteris cunpatis et den-
tatis vel linearibus et integris; foliis caulinis remotis sessilibus
pinnato-lobatis semlamplexicaulibusque sursum scusim reductis;
inflorescentiis terminalibus corymb
1 cm. altis radiatis, ligulis flavis ; involucris campanjilatis minute
caJyculatis; involucri squamis circiter 21 lineari-lailceolatis 6.5-
8 mm. longis acutis glabris maturitate retrorsis; floribus disci
numerosis; achseniis glab
Specimen examined:
Arizona: without defmite locality, coll. of 1883, H. H. Rushy
i 75 (Gray Herb
Professor Rusby's plant was referred by Dr. ' Gray to S
arizonicus Greene, but from the very large root, the sublyrate
smooth and even somewhat glaucous radical leaves, and nearlj
naked stem it seems amply distinct. |
Senecio (§ Tomentosi) oreophilus Greenm. Monogr. Senecio
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
S. neo-vwxicanus Gray, Proc. Am. Acad. 19: 55. J883, in part
Syn. Fl. i^: 392. 1884, in part, as to plant of Greene.
Herbaceous perennis iuventate ubique tomentulosus deniqu
plus minusve glabratus; caule tereti erecto striato 2-3 dm
subnudo 2-3-bracteato; foliis rosulatis petiolatis oblon
obi an
cm
0.7-2.5 cm. latis supra mediam partem crenato-dentatis basi
in petiolum sensim angustatis integriuscuhs juventate utrinque
albo-tomentulosis mox glabratis; bracteis cauUnis linearibus
apice basique parum amphatis dentatisque; inflorescentiis laxe
corymboso-cymosis usque ad 1 dm. diametro; capituUs 10-12
mm. altis calyculatis radiatis; involucris campanulatis basi
tomentulosis ceteris glabris; involucri squamis plerumque 21
[Vol. 1
2G8 ANNALS OF THE MISSOURI BOTANICAL GARDEN
lanceolatis G.5-8 mm. longis acutis; flosculis liguliferis ca. 12,
ligulis oblongis 8 mm. longis 3 mm. latis 4-5-nerviis; floribus
disci iiumerosis ca. 50; achseniis in angulis sursum hispidulis.
Specimen examined:
New Mexico: Pinos Altos Mountains, 6 Alay, 1880, Edward Lee
Greene (Gray Herb.), type.
A plant similar in habit to S. neo-mexicanus Gray, to which
Dr. Greene's specimen was referred by Professor Gray in estab-
lishing that species. A careful study of all the original material,
which has been made possible through the courtesy of Dr. B.
L. Robinson, has shown that the S. neo-mexicanus of Dr. Gray
consisted of at least three recognizably distinct forms of which
Wright's No. 1415, as the first specimen cited, must be taken as
the type. With the Wright plant several specimens at hand are
almost the exact counterpart. Tte Greene plant in question,
namely S. oreophilus, differs in several important particulars,
notably in its essentially naked stem, oblong-cuneate leaves
with subentire or sinuate-dentate margin, and a marked ten-
dency for the foHage to become glabrous with age.
Senecio (§ Tomentosi) oreopolus Greenm. Monogr. Senecio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
Plate 11.
Herbaceus perennis ubique albo-tomentosus; caulibus cses-
pitosis simplice vel ramosis 0.8-3 dm. altis; foliis inferioribus
petiolatis ovato-ellipticis vel elliptico-lanceolatis vel rarius sub-
obovatis 0.8-3.5 cm. longis 5-18 mm. latis obtusis vel supra
mediam partem paucidentatis basi abrupte vel longe cuneatis
integriusculis juventate utrinque albo-tomentosis ietate supra
paululo subinde glabratis, petiolatis 1-6.5 cm. longis, fohis
supremis grosse reductis petiolatis vel sessilibus integris vel
rarius irregulariter dentatis basi sxpe cxpansis et subauricu-
laribus; inflorescentiis corymboso-cymosis; capitulis plerumque
ca. 1 cm. (8-14 mm.) altis radiatis parce calyculatis; involucri
squamis plerumque 13 (9-13) lanceolatis vel lineari-lanceolatis
5
glab
liguliferis 5-13; floribus disci 20-30; pappi setis albis bracteis
involucri longioribus; acha^niis 3-3.5 mm. longis glabris.
1914]
GREENMAN — ^NORTII AMERICAN SENECIONE^ 269
4P
Specimens examined:
California : Rock Creek Canon, Basin of the Upper Kern Kiver,
Tulare Co., altitude 3050 m., July, 1904, H. M. Hall &
H. D. Bahcock, 5526 (Gray Herb.), type; Natural Bridge,
Volcano Creek, Basin of the Upper Kern River, altitude
2285 m., July, 1904, H. M. Hall & H. D. Bahcock, 51^33
(Gray Herb.); gravelly slopes, Little Kern River, altitude
3045-3350 m., April-September, 1897, C. A. Purpus, 52W
(Gray Herb, and Mo. Bot. Card. Herb.); Castle Peak,
near the highest point, altitude 2740 m., 5 August, 1903,
A. A. Heller, 7102 (Gray Herb, and Mo. Bot. Gard. Herb.);
Sierra Nevada, coll. of 1875, John Muir, U52 (Mo. Bot.
Gard. Herb.) ; near the summit of Silver Mountain, altitude
3350 m., coll. of 1863, W. H. Brewer, 2050 (Gray Herb.);
Ebbett's Pass, T^. H. Brewer, 2005 (Gray Herb.); Sonora
Pass, W. H. Brewer, 2686 (Gray Herb.) ; Mono Pass, coll.
of 1866, H. N. Bolander, 6I40 (Gray Herb.).
Nevada: Mt. Rose, Washoe Co., altitude 3200 m., 26 August,
1911, A. A. Heller, 9882 (Mo. Bot. Gard. Herb.).
Forma aphanactis, forma nova.
Caulis circiter 1 dm. altus; foliis petiolo incluso 1.5-2.5 cm.
longis 5-7 mm. latis; capitulis discoideis.
Specimen examined:
California: mountain peak near Sonora Pass, altitude 3200 m.,
coll. of 1863, W. H. Brewer, 1905 (Gray Herb.), type.
Senecio (§ Tomentosi) Wrightii Greenm. Monogr. Senccio,
pt. 1, 24. 1901; in Engl. Bot. Jahrb. 32: 20. 1902, nomen.
S. fastigiatus Gray, PL Wright, ii. 99. 1853, not Nutt.
Herbaceus perennis ubique subtomentosus; caule erecto 1-4
dm. alto foliato; foliis oblongo-oblanceolatis vel lanceolatis
indivisis et integris vel supra mediam partem paucidentatis
juventate albo-tomentosis plus minusve glabratis, inferioribus
basi integriusculis in petiolum sensim angustatis, eis caulinis
sessilibus basi ssepius ampliatim et irregulariter dentatis
amplexicaulibusque ; inflorescentiis terminalibus subcorymboso-
cymosis multicapitatis; capitulis 8-10 mm. altis minute calycu-
latis radiatis; involucris campanulatis basi subincrassatis
tomentosis, bracteis involucri plcrumque 13 lanceolatis 5-7 mm.
londs acutis tomentulosis ; flosculis liguliferis 6-8, ligulis anguste
[Vol. 1
270 ANNALS OF THE MISSOHIII BOTANICAL GARDEN
oblongis ca. 8 mm. longis 4-5-nerviis; floribus disci ca. 30;
achaeniis glabris.
Specimens examined :
New Mexico: ravines hefwpon fhn pnm^Ar
S
): ravines between the copper mines and the Mim-
bres, October, 1851, Charles Wright, 1289 (Gray Herb,
and Mo. Bot. Gard. Herb.), type; Santa Rita del Cobre,
22 September, 1880, E. L. Greene (Mo. Bot. Gard. Herb.);
among spruce, Lookout Mine, Sierra Co., altitude 2680
nh, 0. B. Metcalfe, 1179 (Mo. Bot. Gard. Herb.).
§ Amplectentes) subauriculatus Grccnm. Monogr
Senecio, pt. 1, 25. 1901; in Engl Bot. Jahrb. 32: 21. 1902,
nomen
Plate 14.
Herbaccus perennis; caule erecto ramoso striato glabro; foliis
in partibus superioribus caulinis anguste lanceolatis 5-15 cm.
longis 0.5-1.5 cm. latis acuminatis acutis integris vel remote
apiculato-dcnticulatis sessilibus et auriculo-semiamplexicauli-
bus vel basi in petiolum sensim angustatis et subdccurren-
tibus membranceis supra glabris juventate subtus floccoso-
tomentosis denique plus minusve glabratis; infloresccntiis ter-
minalibus laxe subcorymboso-cymosis; pedunculis bracteatis,
bracteis lineari-attenuatis; capitulis radiatis 12-14 mm. altis
heterogamis; involucris campanulatis calyculatis albo-floccoso-
tomentulosis, bracteolis calyculatis linearis acutis suberoso-
squamis plerumque 21 lineari-lanceolatis
cm
13, ligulis oblongis flavibus; floribus disci numerosis (50-60);
pappi setis albis; achaeniis pubescentibus.
Specimen examined :
Mexico: State of Oaxaca, mountains southeast of Miahuatlan,
altitude 2750-3170 m., coll. of 1895, E. W. Nelson, 2526
(Gray Herb.), type.
A well marked species related to S. Warszewiczii A. Br. &
Bouchd and to S. prionopterus Rob. & Greenm.
Senecio (§ Mulgedifolii) alatipes Greenm. Monogr. Senecio,
pt. 1, 25. 1901; in Engl. Bot. Jahrb. 32: 21. 1902, nomen.
Herbaceus perennis ubique glabrus; caule tereti striato erecto
1 m. vel ultra alto; foliis parte inferiori ignotis, eis cauUnis
petiolatis vel sessilibus amplexicaulibusque oblongo-ovatis vel
oblongo-lanceolatis 0.5-1.5 dm. longis 2-5 cm. latis acutis vel
1914]
GREENMAN — NORTH AMERICAN SENECIONE^ 271
acuminatis indivisis vel subpandurifortnibus utrinque glabris
subtus pallidoribus, margine irregulariter calloso-dentatis; peti-
olis usque ad 12 cm. longis anguste alatis; inflorescentiis
terminal ibus^paniculatis; capitulis 8-10 mm. altis discoideis
20-25-floris ; involucris anguste campanulatis calyculatis glabris;
involucri squamis plerumque 13 Hneari-lanceolatis acutis penicil-
latis ca. 6 mm. longis; achseniis striatis glabris.
Specimen examined:
Mexico: State of Chiapas, between Teneapa and Yajalon, alti-
tude 900-1520 m., 13 October, 1895, E. W. Nelson, 3277 (U.
S. Nat. Herb., fragments and tracing in Gray Herb.), type.
498
Senecio (§ Mulgedifolii) callosus Schz. Bip. in Flora 28:
S. eximius Hemsl. Biol. Cent.-Am. Bot. 2: 239. 1881, as to
synonomy.— >§. doralophyllus Hemsl. l. c, in part, as to Bour-
No. 1086, not Benth.— >Sf. viejensis and S. latipes Greenm.
Jahrb
Monogr. Senecio, pt. 1, 25.
1902, nomen.—Cacalia Toluccana DC. Prodr. 6: 328. TsS?.—
C. prenanthoides Gray, Proc. Am. Acad. 19: 53. 1883, in part,
as to Bourgeau's No. 1086, not liBK.— Erechthites runcinaia
Hemsl. Biol. Cent.-Am. Bot. 2: 234. 1881, in part, as to Bour-
geau's No. 1086, not DC.
Herbaceus perennis ubique glabrus vel sparsissime tomen-
tellus; caule tereti erecto circiter 1 m. alto striato plus minusve
purpurascenti; foliis radicalibus et eis caulinis infimis petiolatis
vel sessilibus amplexicaulibusque runcinato-pinnatifidis, lobis
remotis, usque ad 4 dm. longis 3-18 cm. latis utrinque glabris
subtus pallidioribus calloso-dentatis, summis sessilibus et auri-
culato-amplcxicaulibus indivisis lanceolato-attenuatis; inflores-
centiis terminalibus laxe paniculatis poly ceph alls; capitulis
discoideis 10-12 mm. altis calyculatis 15-34-floris; involucri
squamis plerumque 13 (8-13) lineari-lanceolatis 8-10 mm. longis
acutis glabris et coroUis plus minusve purpurascentibus ; pappi
setis albis; acha^niis striatis glabris.
Specimens examined:
Mexico: State of Mexico, Desierto Viejo pres Mexico, Bourgeau,
1086 (Gray Herb, and Berlin Herb.) ; near Guapimalpam,
coll. of 1855, Schaffner (Gray Herb.); fir woods, Sierra de
las Cruces, 11 December, 1892, C. G. Pringle, 5313 (Gray
[Vol. 1
272 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Herb.) ; Sierra de las Cruces, altitude 3350 m., 11 February,
1899, C, G. Pringle, 7709 (Mo. Bot. Gard. Herb.); Mt.
Ixtaccihuatl, altitude 2430-3350 m., C. A. Purjpus, 100
(Gray Herb.) ; fir forests, Mt. Ixtaccihuatl, altitude 3350-
3050 m., February, 1903, C. A. Purpus, 1^5 (Mo. Bot.
Gard. Herb.). State of Vera Cruz, Las Vigas, near Jalapa,
2 December, 1903, C. G. Pringle, 11S69 (Gray Herb.),
forma. State of Oaxaca, without definite locality, Cuming
(Gray Herb.). State of Colima, coll. of Jan. 9-Feb. 6,
1891, Dr. Edward Palmer, 1145 (Gray Herb.), distributed
DC
>j
The examination of a large suite of herbarium specimens,
particularly in the light of recently acquired material, has led
the writer to place a somewhat different interpretation on this
species than formerly; hence, a brief description is here given
and a few specimens from widely distributed exsiccati, well
illustrating the species, are cited.
Senecio (§ Mulgedifolii) Coulteri Grecnm. Monogr. Senecio,
pt. 1, 25. 1901; in Engl. Bot. Jahrb. 32: 21. 1902, nomen.
Cacalia runcinata Less. Linna^a 5: 1G2. 1830, not HBK.
Herbaceus perennis; caulibus erectis 3-6 dm. altis striatis
paulo tomentulosis plus minusve purpurascentibus ; foliis
inferioribus petiolatis runcinato-pinnatifidis usque ad 3 dm.
longis 1.5-6 cm. latis supra glabris subtus arachnoideo-tomentu-
losis ina^qualiter et obtuse calloso-dentatis, foliis supcrioribus
gradatim reductis sessilibus amplexicaulibusque; inflorescentiis
terminalibus subcorymboso-cymosis; capitulis numerosis dis-
coideis ca. 1 cm. altis brevi-calyculatis; bracteis involucri ple-
rumque 13 lanceolatis acutis 8 mm. longis glabris et purpura-
scentibus; floribus disci 30-40; achocniis glabris.
Specimens examined:
Mexico; State of Vera Cruz, Real del Monte, Dr. Thomas
Coulter, 429 (Gray Herb.), type, C. Ehrenhcrg, 381 (Berlin
Herb, and Gray Herb.) ; Mt. Orizaba, Schiede, 363 (Berlin
Herb.). State of Mexico, on Nevada de Toluca, 15 October,
1903, J. N. Rose & J. N. Painter, 7940 (U. S. Nat.
Herb, and Gray Herb.).
Senecio (§ Mulgedifolii) iodanthus Greenm. Monogr. Senecio,
Jahrb
Plate 12.
1914]
GREENMAN — NORTH AMERICAN SENECIONEiE 273
Herbaceus perennis; caulibus 5-9 dm. altis foliaceis striatia
glabris plus minusve purpurascentibus; foliis inferioribus pler-
umque lyrato-pinnatifidis oblongo-lanceolatis 1.5-3 dm. longis
3.5-9 cm. latis acutis vel acuminatis sinuato-calloso-dentatis
supra glabris subtus juventate arachnoideo-tomentulosis et
ssepe crispo-puberulentis fere glabratis, foliis superioribus sur-
sum gradatim reductis sessilibus amplexicaulibusque; inflores-
centns racemoso-paniculatis 2-5 dm. longis 0.3-1.2 dm. latis;
capitulis 10-12 mm. altis discoideis calyculatis; bracteis involucri
circiter 13 lanceolatis 8 mm. longis acutis vel obtusis penicillatis
glabris vel sparse puberulentis purpurascentibus; floribus disci
ca. 24; pappi setis albis quam corolla brevioribus; coroUis albisvel
purpurascentibus; achseniis glabris.
Specimens examined:
Mexico: State of Mexico, in pine woods, Nevada de Toluca,
altitude 3000-3600 m., 26 September, 1892, C. G. Pringle,
4302 (Gray Herb, and Mo. Bot. Gard. Herb.), type.
State of Morelos, Tres Marias Mts., altitude 2895 m., 5
November, 1903, C. G. Pringle, 11498 (Gray Herb.).
This species is closely related to S. CouUeri Greenm. but
differs in having a smooth and more leafy stem, nearly glabrous
leaves, and distinctly racemose-paniculate inflorescence.
Senecio purpurascens Klatt, Leopoldina, Heft 24, p. 126.
1888.
Var. fossanervius Greenm. Monogr. Senecio, pt. 1, 25. 1901;
in Engl. Bot. Jahrb. 32: 21. 1902, nomen.
Formse typicse habitu simili; foliis inferioribus petiolatis, peti-
olo inclusoj usque ad 11 cm. longis 1.5 cm. latis sinuato-dentatis
vel ad basin sublyratis supra glabris fossanerviis subtus tomen-
tellis et in nerviis pilosis; involucri squamis fere glabris.
Specimen examined :
Mexico: without definite locality, E. W. Nelson, 1308 in part
(U. S. Nat. Herb., fragments in Gray Herb.), type.
Senecio (
osi) carnerensis Greenm. Monogr. Sene
Engl. Bot. Jahrb. 32: 21. 1902, nomen.
Perennis basi suffrutescens ubique plus minusve 1
ule tereti erecto simplici vel ramoso; foliis indivisis
1 sessilibus lanceolatis vel oblanceolatis 1.5-5
cm
ad 1 cm. latis acutis denticulatis iuventate utrinoue tomentosia
[Vol. 1
F
274 ANNALS OF THE MISSOURI 'BOTANICAL GARDEN
supra plus minusve glabratis subtus persistenter albo-tomen-
tosis, superioribus subauriculatis ; inflorescentiis terminalibus
brevi-calvculatis radiati
bracteis
■labris vel narce tomentulosis ; flosculis
umque 8, ligulis flavis 4-nerviis; floribus disci 30-40 acliaeniis
sursum brevi-sericeo pubescentibus.
Specimen examined:
Mexico: State of Coahuila, mountains, Carneros Pass, altitude
3050 m., 8 September, 1889, C. G. Pringle, 2857 (Gray
Herb., photograph in Mo. Bot. Gard. Herb.), type.
This species was originally referred to S. longilobus Benth.,
but it is more closely allied to S. siocchadiformis DC. and S.
Picridis Schaucr; it is readily separated from both these species
by having fewer involucral bracts, short, appressed and
black-tipped bracteoles suggesting those of S. vulgaris L.
Senecio (§ Suffruticosi) filicifolius Grcenm. Monogr. Sene-
cio, pt. 1, 25. 1901; in Engl. Bot. Jahrb. 32: 21. 1902; Contr.
U. S. Nat. tierb. 16: 19. 1912, nomen.
Herbaceus perennis (?) erectus ramosus 1.5-4 dm. altus ubique
glabrus; caule tereti ad basin plus minusve lignescenti; ramis
ramulisque striatis stramineis; foliis sessilibus vel subalato-peti-
olatis pectinato-pinnatifidis 1.5-8 cm. longis 1-6 cm. latis; seg-
mentis linearis attenuatis acutis; inflorescentiis subcorymboso-
cris cam
ligocephalis; capitulis ca. 1
panulatis calyculatis; involucri squamis plerunique 21
bracteolis calyculatis duplo longioribus lineari-lanceolatis acutis
glabris vel juventate parce tomentulosis mox glabratis; flosculis
liguliferis ca. 12, ligulis flavis; floribus disci 50-60; pappi setis
albis; achaniis sursum sericeo-hispidulis.
Specimens examined:
Arizona: Valley of the Santa Cruz River, 11 May, 1881, C. G.
Pringle, 316 (Gray Herb.), type; Tucson, 12 March, 1892,
/. Tf. Tourney, 708 (Gray Herb.); Tempo, coll. of 1892,
Ganong & Blaschka (Gray Herb.) ; Hart's Ranch, 17 miles
south of Tucson, 11 April, 1903, J. J. Thornber, 436 (Mo.
Bot. Gard. Herb.); Ft. Huachuca, coll. of 1894, Maj. T. E.
Wilcox (Mo. Bot. Gard. Herb.); open caiions, San Francisco
Mts., April, 1887, H. H. Rushy, 2H in part (Mo. Bot. Gard.
Herb.").
GBBENMAN— NORTH AMERICAN SENECIONE^ ^/O
Mexico: Sandy plains near Altar, State of Sonora, 4 April,
1884, C. G. Pringle (Gray Herb.).
This species has been hitherto included with S. Douglasii
DC. from which it differs in being essentially glabrous through-
out, in having usually more numerous and shorter lateral
leaf-segments, fewer, shorter, and less conspicuous calyculate
bracteoles.
4
Senecio (§ Suffruticosi) teliformis Greenm. Monogr. Senecio,
pt. 1, 26. 1901; in Engl. Bot. Jahrb. 32: 22. 1902, nomen.
Herbaceus perennis; caule erecto tereti superne striato
stramineo floccoso-tomentoso plus minusve glabrato; fohis su-
premis sessilibus lanceolato-attenuatis 3-6 cm. longis ad basin
ampliatis usque ad 1.5 cm. latitudine semiamplexicaulibusque
supra juventate floccoso-tomentulosis plus minusve glabratis
subtus persistenter albo-tomentosis, margine dentatis vel denti-
culatis revolutisque ; foliis inferioribus ignotis; inflorescentiis
terminalibus corymboso-cymosis multicapitatis bracteatis floc-
coso-tomentulosis; capitulis 8-10 mm. altis radiatis calyculatis,
bracteolis calyculatis lineari-attenuatis conspicuis subflaccidis
floccoso-pubescentibus; involucri bracteis plerumque 21 lineari-
lanceolatis 5-6 mm. longis acutis glabris penicillatis; flosculis
liguliferis sa^pius 8, ligulis oblongis 5-6 mm. longis flavis;
floribus disci ca. 40 quam bracteis involucri longioribus, pappi
setis albis; acha^niis sursum adpresso-scriceo-pubescentibus
maturitate 3 mm. longis.
Specimen examined:
Mexico: State of Oaxaca, mountains of Telixtlahuaca, altitude
2500 m., 10 December, 1894, Rev, Lucius C. Smith, 367
(Gray Herb., photograph and fragments in Mo. Bot. Gard.
Herb
of the
to the writer, nevertheless it evidently belongs to the section
Suffruticosi and appears to be most closely related to S. Picridis
Schauer and S. alvarezensis Greenm. From the former it differs
by the usually broader base of the upper stem leaves, more
numerous heads and conspicuous bracteoles, while from the
latter it is readily separated on foliar characters
Senecio (^ Palmatinervii) albonervius Greenm. Mon
Senecio, pt. 1, 26. 1901; in Engl. Bot. Jahrb. 32: 22. 1902,
nomen.
[Vol. 1
276 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Arborescens 2-4 m. altus; caule tereti prime albo-tomcntuloso
maturitate glabrato et cortice brunnco tecto; foliis petiolatis
basi palmatinerviis late ovatis 3-5 cm. longis latisque simiato-
5-11-lobatis remote calloso-mucro-dcnticulatis basi cordatis
tomentiilosis plus minusve glabratis
umque
in-
m nerviis persistenter albo-tomentulosis, petiolis
3-10 (usque ad 14 cm.) longis; inflorescentiis terminalibu
culatis multicapitatis; capitulis 10-12 mm. altis radiatis,
yolucris anguste campanulatis vel subcylindricis brevicalyculatis;
involucri squamis circiter 8 lineari-lanceolatis vol oblongis ob-
tusis 5-6 mm. longis glabris vel i)arce tomentulosis; flosculis
liguliferis plerumque 5, ligulis 5-7 mm. longis flavis 4-nervatis,
pappi setis tubo corollse longioribus; floribus disci 8-10; ach-
seniis glabris.
Specimens examined ;
Mexico: State of Mexico, Valley of Temascaltepec, April, 1831,
Schiede (Berlin Herb, and Gray Herb.), type; open woods,
Ixtaccihuatl, altitude 2430-3350 m., March-July, 1903,
C. A. Purpus, 201 (Gray Herb, and Mo. Bot. Gard. Herb.).
State of Vera Cruz, Mineral del Monte, Ehrcnhcrg, 3^
(Berlin Herb, and Gray Herb.). State of Morelos, Sierra
de Tres Marias, altitude 3050 m., 15 April, 1904, C. G.
Pringle, 8903 (Gray Herb, and Mo. Bot. Gard. Herb,
State of Michoacan, north slope of Mt. Tancitaro, altitude
2280-3200 m., 24 February, 1903, E. W. Nelson, 6904
(U. S. Nat. Herb, and Gray Herb.).
^ The broadly ovate, shallowly sinuate-lobed leaves with per-
sistent white tomentum on the veins of the upper leaf-surface,
together with a terminal many-headed panicle and yellow ray-
flowers, render this species distinct and easily recognized among
all those of the palmately veined section to which it belongs,
Senecio angulifolius DC., var. ingens, var. nov.
Habitu et foliis forma; typicae; inflorescentiis compactis
pauci- vel multi-capitatis, bracteis bracteolisque perconspicuis;
capitulis 1.5-2 cm. altis 40-45-floris radiatis vel discoideis.
Specimens examined :
Mexico: Mt. Ixtaccihuatl, above timber line, March- July,
1903, C. A. Purpus, 193 (Mo. Bot. Gard. Herb.), type;
rocky slopes, Mt. Ixtaccihuatl, altitude 5790-6090 m'
1914]
GREENMAN — NORTH AMERICAN SENECIONE^ 277
November, 1905, C. A. Purpus, 1517 (Mo. Bot. Gard.
Herb.). State of Puebla, Mt. Orizaba, near Chalchicomula,
25 February, 1892, Jared G. Smith, 473 (Mo. Bot. Gard.
Herb.).
On account of the conspicuous more or less foliaceous bracts
of the inflorescence S. angulifolius DC. is a very characteristic
species and is almost always recognized without difficulty.
There is, however, a considerable variation in the size of the
heads and in the number of flowers of the disk, as well as in the
degree of development of the ray-flowers. In fact the latter
may be well developed, more or less reduced, or entirely absent.
The extremely large headed form, which is well exemplified by
the specimens cited above, seems well worthy of varietal rec-
ognition. Doctor Purpus's No. 1517 is somewhat intermediate
between the species and the variety.
Senecio (§ Palmatinervii) brachyanthus Grccnm. Monogr.
Senccio, pt. 1, 2G. 1901; in Engl. Bot. Jahrb.32 : 22. 1902, nomen.
Verisimiliter frutex; caule tereti cortice brunneo tecto juven-
tatc hirtcllo-puberulento glabrato; foliis longipetiolatis sub-
peltatis palmatinerviis suborbicularis circiter 7-lobatis mem-
branaceis utrinque parce hirtellis subtus pallidioribus mucro-
denticulatis, petiolis usque ad 13 cm. longis minute puberulentis;
inflorescentiis terminalibus subglanduloso-hirtellis; capitulis sub-
cylindricis 10-12 mm. altis heterogamis; involucri bracteis 8
lanceolatis 8-10 mm. longis acutis vel obtusis plus minusve
purpurascentibus extus subglanduloso-hirtellis; flosculis fem-
ineis 5 multum reductis, ligula nulla, tubo gracili squamis invol-
ucri breviore; floribus disci 8-10; pappi setis albis; achaeniis
glabris.
Specimen examined:
Mexico: State of Guerrero, between Ayusinapa and Petatlan^
altitude 1540-2155 m., E. W. Nelson, 2137 (Gray Herb.
and U. S. Nat. Herb.), type.
The leaves and reduced ray-flowers of this species are similar
to those of S. cordovensis Hemsl., but the character of the
involucre indicates a closer relationship with S. chapalensis
Watson.
Senecio (§ Palmatinervii) chapalensis Watson, Proc. Am.
Acad. 25: 155. 1890.
^IV> ANNALS OF THE MISSOURI BOTANICAL GARDEN
Var. areolatus Greenm. Monogr. Senecio, pt. 1, 26. 1901;
in Engl. Bot. Jahrb. 32: 22. 1902, nomen.
A forma typica recedit foliis utrinque glabratis subtus areo-
latis, petiolis usque ad 15 cm. longis plus minusve purpur-
ascentibus ; flosculis liguliferis granditer reductis.
Specimen examined:
]\Iexico: State of Morelos, on shaded bluffs of a wet canyon
above Cuernavaca, altitude 1980 m., 15 February, 1899,
C. G. Pringle, 8010 (Gray PIcrb. and Mo. Bot/card!
Herb.), type.
Senecio (§ Palmatinervii) Chrismarii Greenm. Monogr.
Senecio, pt. 1, 2G. 1901; in Engl. Bot. Jahrb. 32: 22. 1902,
nomen.
Frutex;^ caule primo parce pubescenti maturitate glabro;
)lns petiolatis palmatinerviis circumscriptione triangulari-
ovatis 7-10 cm. longis 5-8 cm. latis hastatis 3-5-lobatis ciliatis
mucro-dcnticulatisque granditer cordatis supra sparse hirtello-
pubcrulentis subtus glabris vcl in nervis puberulentis, lobiis
mucronato-acutis; petiolis gracilibus 4-9
f
cm
bus remote bi
tellis vel glabris; inflorescentiis terminalibus laxe paniculatis
paucicapitatis dense glanduloso-pubcrulentis, pedunculis gra-
icteatis; capitulis 1.2-1.5 cm. altis discoideis
paucicalyculatis; involucri squamis sa^pius 8 lanceolato-oblongis
ca. 1 cm. longis acutis penicillatis extrinsecus hirtello-pubcru-
lentis plus minusve purpurasccntibus interioribus scarioso-
marginatis; floribus disci plerumque 20 involucri bracteis
longioribus; pappi setis albis; achaniis glabris.
Specimen examined :
Mexico: without definite locality, Chrismar (Berlin Herb.,
tracing and fragments in Gray Herb.), type.
Tlie afhnity of this species is with S. hedercefolius Hemsl., S,
anisophyllus Klatt, and S. alicnus Robinson & Scaton. From
the first two it differs in having deeply cordate leaves with more
or less reflexcd lateral lobes, and from the last it is readily sep-
arated by the deeply cordate loaves and absence of peltation.
Senecio (§ Palmatinervii) hypomalacus Greenm. Monogr,
pt. 1, 20. 1901; in Engl. Bot. Jahrb. 32: 22. 1902,
Sen
nomen.
Plate 10
1914]
GREENMAN — NORTH AMERICAN SENECIONE^ 279
Fnitex erectus; caule tereti primo dense sordid o-puberu-
lento, saepissime lenticellis intermixtis, maturitate cortice
brunneo tecto; foliis petiolatis vel supremis sessilibus circum-
scriptione ovato-rotundatis vel ovato-oblongis palmato-3-5-
nerviis distincte 5-11-lobatis supra crebe crispo-hirtellis subtus
lanato-tomentosis basi cordatis vel subtruncatis, margine sinua-
tis calloso-denticulatis ciliatis; petiolis usque ad 6 cm. longis;'
inflorescentiis terminalibus paniculatis polycephalis subglandu-
loso-hirtellis; capitulis 10-12 mm. altis parce calyculatis radiatis;
bracteis involucri plerumque 8 (non-nunquam 7) oblongis vel
subobovatis 5-6 mm. longis obtusis vel acutis extus crebe sub-
glanduloso-hirtellis, interioribus late scarioso-marginatis; fios-
culis femineis liguliferis, ligulis anguste oblongis 5-6 mm. longis
flavis; floribus disci circiter 10 (7-13) quam involucrum bis tanto
fere longioribus; pappi setis albis; achseniis glabris.
Specimens examined :
Mexico: State of Oaxaca, mountains of Telixtlahuaca, altitude
2375 m., 10 December, 1894, Rev. Lucius C. Smith, 368
(Gray Herb., photograph and fragments in Mo. Bot. Gard.
Herb.), type; Sierra de San Felipe, altitude 2130-2440 m.,
17 November, 1894, Charles L. Smith, 210 (Mo. Bot. Gard.
Herb.); Cerro de San Felipe, altitude 1900 m., 25 September,
1895, C. Conzatti, 119 (Gray Herb.).
This species is related to ;S. oaxacanus Hemsl., but differs
from it in having distinctly lobed leaves which are thicker in
texture, densely subglandular-hirtellous above and soft tomen-
tose beneath; moreover, the leaf-margin of S. hypomalacus is
markedly sinuate and the lobes show a tendency to become again
lobate. C. and E. Seler's No. 1581 from Tillantongo, which
has been referred to S. oaxacanus Hemsl., is somewhat intermedi-
ate between the two species, but it has the leaf-outline and
thinner texture of Mr. Hemsley's species.
Senecio (§ Palmatinervii) Kerberi Greenm. Monogr. Senecio,
pt. 1, 26. 1901; in Engl. Bot. Jahrb. 32: 22. 1902, nomen.
Herbaceus robustus perennis usque ad 3m. altus; caule
tereti erecto glabro vel parce tomentulosb; foliis petiolatis pal-
mato-5-7-nerviis ovato-oblongis 5-10 cm. longis 5-8 cm. latis
5-7-lobatis carnoso-denticulatis reticulato-venosis supra sparse
hirtellis subtus subarachnoideo-tomentulosis, lobis obtusis vel
2
280 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Voii. 1
subrotundatis et mucronato-acutis; pctiolis 2-2.5 cm. longis;
inflorescentiis terminalibus paniculatis niulti-capitatis pubes-
centibus, pedunculis minute bracteatis; capitulis 7-8 mm. altis
radiatis; involucris cainpanulatis minute calyculatis fere glabris;
involucri squamis 13 lineari-lanceolatis vel lanceolato-oblongis
4.5-5 mm. longis acutis glabris; flosculis femineis 5 ligulifcris,
ligulis oblongis 4-5 mm. longis flavis; floribus disci ca. 14, pappi
sctis albis; achscniis glabris.
Specimen examined:
Mexico: "Tromptcro, Mesa del Arrero," 21 November, 1880,
Kerher, 94 (Berlin Herb., fragments and tracing in Gray
Herb.), type.
This species is known at present from a single specimen in
the Royal Botanical Museum of Berlin. From this specimen
the writer was permitted, as in a number of other cases, while
making a study of the genus several years ago, to make a tracing
and take fragments for the Gray Herbarium of Harvard Uni-
versity. The species is related to S. Hariwegi Benth. and S.
rcglcnsis Greenm., but from these and from other species of the
section Palmaiinervii to which it belongs, it is readily distin-
guished by the somewhat elongated more or less fan-shaped
and bluntly lobed leaves.
Senecio (§ Palmatinervii) velatus, sp. nov. Plate 13.
Frutex; caule tereti carnoso ramoso ad apicem sordido-tomen-
toso cetero glabro in sicco cortice brunneo tecto; foliis petiolatis
palmato-7-nerviis circumscriptione ovato-rotundatis ca. 10 cm.
longis latisque anguIato-7-9-lobatis membranaceis integris ju-
ventate utrinque plus minusve albo-tomentosis subtus persis-
tenter arachnoideo-tomentulosis, lobis triangulari-ovatis mu-
cronato-acutis; petiolis ca. 8 cm. longis floccoso-pubescentibus;
inflorescentiis terminalibus dense cymoso-corymbosis minute
bracteatis multicapitatis glabris vel in axillis ramulorum floccoso-
tomentulosis; capituhs ca. 1.5 cm. altis radiatis; involucri sub-
cylindrici squamis ssppius 8 lanceolato-linearis vel lanceolato-
oblongis 7-10 mm. longis acutis vel obtusis; flosculis ligulifcris
3-5, ligulis anguste oblongis ca. 1 cm. longis; floribus disci 6-7,
pappi setis albis; achaeniis glabris striatis.
1914]
GREENMAN — NORTH AMERICAN SENECIONE^ 281
Specimen examined :
Mexico : State of Jalisco, on bluffs of barranca, near Guadalajara,
20 May, 1891, C. G. Pringle, 5160 (Gray Herb., photograph
and fragments in Mo. Bot. Gard. Herb.), type.
The writer has withheld publication of this species for several
years with the hope that additional material might be secured.
Mr. Pringle's specimen, from which the above description is
drawn, is in the Gray Herbarium and consists of a terminal
portion of a flowering stem and two detached leaves. In stem
and inflorescence characters it corresponds very well with typical
specimens of S. prcecox DC. except that the terminal portion
of the stem and branches are covered with a tawny pubescence,
not glabrous as is usually the case with the DeCandollean spe-
cies. On account of the similarity of stem and inflorescence
and because of the detached leaves the plant has been referred
doubtfully to the peculiarly characteristic and well known S.
prcecox DC.
The extreme care with which Mr. Pringle prepared his plant
material and the fact that the leaves on the specimen under
consideration, although detached from the stem, accord with the
type of foliage of the section Palmatinervii lead me to believe
that we have to deal in the present case with an unrecorded
species related to but distinct from S. prcecox DC, and in all
probability one of limited geographical distribution.
Senecio Klattii, nom. nov.
S. rosews Klatt, Ann. k. k. Naturhist. Hofmus. Wienp: 366.
1894. not. S. roseus Schz. Bin. in Flora 28: 498. 1845.
282 ANNALS OF THE MISSOURI BOTANICAL GARDEN
fVoL, 1, 1914]
Explanation of Plate
PLATE 10
Senecio hypomalacus Grconm.
Mexico
From the type specimen, Rev. Lucius C. Smith No. 368, in the Gray
Herbarium of Harvard Uuivertiity.
Anv. Mo. Box. Gard., Vol. 1, 1914
Plate \0
GREENMAN— NORTH AMERICAN vSENEClOXEAE
COCKAVMK. BOSTON.
284
(Vol. I, 1914]
ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 11
Senecio oreopolus Greenm.
California
From the type specimen, Hall and Babcock No. 5526, in the Gray
Herbarium of Harvard Univeraity.
AiN-N. Mo. 150T. GakDm Vol. 1. I*.tl4
1*1. ATK 11
'''l^^x
. >
grki:njman
NORTH AMERICAN SENrX'IONEAE
COCKAYNE, IJOSrON
IVoL. 1, 1914]
286 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 12
Senecio iodanthus Greenm.
Mexico
om the type specimen, Pringle No. 4302, in the Gray Herbarium
Harvard Universitv.
A^'N. Mo. lioT. (tAUI>., Vol. 1, 1914
Platk V2
)
\
GREEN MA>J
T
NORTH AMERICAN SENECIONEAE
COCKAV^K, liOSXON
[Vol. 1. 1914}
288 ANNALS OF THE MISSOUIU BOTANICAL GARDEN
Explanation of Plate
PLATE 13
Senecio velatus Greenm.
Mexico
From the type specimen, Pringle No. 5160, in the Gray Herbarium of
Harvard University.
Ann. Mo. But. Gard., Vol. 1, 1914
rLATE 13
4 >■ <w'',';'-'*'^
GREENMAN — NORTH AMERICAN SENECIONEAE
COCKAYNE, JIOSTOX
(Vol. 1, 19141
290 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 14
Senecio suhauriculatus Greenm.
Mexico
iimen
barium of Harvard University.
1-
Ann. Mo. Bot. Gaud., Vol. 1, 1914
I'LATJ': 14
^«"-
G R E L N M A N
NORTH AMERICAN SENECIONEAE
COCKAYNK, liOSTON
A STUDY OF THE PHYSIOLOGICAL RELATIONS OF
SCLEROTINIA CINEREA (BON.) SCHROTER
T
J. 8. COOLEY
W
Formerly Rufiis J, Lackland Fellow in the Henry Shaw School of Botany of
Washington University
Introduction
This paper reports the results of an experimental study regard-
ing certain physiological activities of the brown-rot fungus of
stone fruits. The investigation concerns itself primarily with
the conditions influencing the penetration and infection of
green and ripe fruits by the fungus in question, the action of
the parasite on the host cell, and the secretion of the enzymes
which act upon the cellulose and pectic substances of the host.
The work was undertaken with the hope of throwing some further
Hght upon the factors concerned in fungous parasitism. Our
present conception of this subject is based upon fragmentary
and, in some respects, contradictory evidence. However, each
year there are acquired new facts, or new applications of known
facts, bearing upon this exceedingly involved and complex
question. An examination into the history of investigations
concerning the interaction of host and parasite shows that the
study of this subject dates back to the work of the pioneers in
plant pathology; modern methods and recent discoveries have,
however, given an added impetus to research along this line. '^
Progress in combating fungous diseases depends not only
upon a familiarity with the life history of the parasite, but more
especially upon an intimate knowledge of the metabolism of the
■m
parasite and the nature of the changes which it induces in the
host. Indeed, many of our recommendations for controlling
parasitic diseases of plants will perhaps be modified when a
more exact knowledge of the interrelations of host and parasite
is gained. Furthermore, a more intimate knowledge of the
physiological aspects of plant pathology will undoubtedly throw
much light on the question of immunity and susceptibility.
We should, of course, like to know more about the factors
favoring or inhibiting parasitic action, as well as the conditions
Ann. Mo. Bot. Gard., Voii. 1, 1914
(291)
;
[Vol. 1
292
ANNALS OF THE MISSOrRI BOTANICAL GARDEN
\
^
\
V
\
\
which influence the infection and the penetration of parasitic
fungi. It would also be interesting to know why some fungi
are so virulent and rapid in their destructive action on the host;
for instance, it would be instructive to know whether it is due
to the secretion of an enzyme, or a toxic substance (e. g., some
acid), or to the disturbance of the osmotic relations of the host
cells, or to some other perhaps unknown factor. For a study of
some of these problems the writer has chosen as the organism
Sclcrotinia cinerea (Bon.) Schroter, the fungus causing the brown
rot of stone fruits. This form is particularly suitable for the pur-
pose since it is a virulent parasite, yet grows well as a saprophyte
— readily lending itself to cultivation in the laboratory.
Historical Review
Space will permit only a brief review of some of the more
important papers dealing with certain aspects of this subject.
Much of the literature that is indirectly concerned with the
problem, or that is fully reviewed or superseded by subsequent
publications, will not be discussed here.
In the period from 1858 to 1878 little experimental evidence
appeared concerning the nature of the action of fungous para-
sites, although several writers make mention of the penetration
of host cells by fungous hyphae. Penetration was then fre-
quently spoken of as merely a process of boring through (^^durch-
bohrung'^) the host tissue, Kiihn (34), as early as 1858, men-
tiQnitxQ:..this fact in a discussion of the potato-bhght fungus.
A few years later, in 1863, de Bary (1) speaks of the penetration
of the host by Peronosj)ora, and further makes mention of this
fact in connection with his work on the rusts (2); again in his
work 'Morphologie und Physiologic der Pilze, Flechten, und
Myxomyceten^ (3) he discusses the penetration of the host, but
says he has no knowledge of the force that causes this boring
into the host tissue.
Hartig (26), in his early work on wood-destroying fungi, as
well as in his later investigations, emphasizes the fact that fungi
are able to destroy cellulose. By a microscopical study of
diseased wood he found that the properties of the latter are
very materially changed by the fungus; he did not, however,
attempt to isolate an enzyme.
■s.
1914]
COOLEY — SCLEROTINIA CINEREA 293
De Bary (4), in 1886, gives us the first important contribu-
tion to our knowledge concerning the action of parasites on host
cells. This author, in his epoch-making research on the fungus
now known as Sderotinia libertiana, reports that the organism
secretes a substance that discolors, plasmolyzes, and finally
kills the host cells. This toxic secretion penetrates the host
cells in advance of the fungus, killing them before they are
actually pierced by the fungous filaments. De Bary was able
to isolate this toxic substance, which he considered as probably
enzyme, and found that it would
the
host tissue similar to that produced by an attack of the fungus
itself. He holds that the fungus will not grow on living tissues,
for it attacks only through a wound and kills the cells in ad-
vance of itself, thus not actually growing upon the living tissue.
The product resulting from the disintegration of the cell wall
of the host was thought to be a sugar that served as food for
the fungus. In this connection de Bary also mentions finding
oxalic acid encrusting the older fungous filaments.
t
The next important paper on the interaction of host and
parasite was that of Marshall Ward (51) pubhshed just two years
after de Bary's work and concerning itself with a species of
Botrytis causing a lily disease. In this excellent piece of work
the author showed that the fungous hyphae on coming in
contact with such solid substances as sections of a lily bulb, or
even a cover glass, secrete from the tips drops of a subs+ance that-
has a very peculiar effect on the host cell. He found that a
water extract of this secretion when applied to sections of a
lily bulb will cause the cell walls to swell and to assume an ab-
I
normal appearance; the middle lamella is first dissolved and
finally the entire cell wall is disorganized. Ward does not con-
sider that this toxic secretion is stimulated by starvation.
Several investigators have held that the penetration of
many fungi is due to chemotropism, i. e., that penetration of
the fungous hyphae is due to some stimulus which the constit-
uents diffusing slowly from within the host cells exert. Biisgen
(16), Miyoshi (39), Behrens (6), Schmidt (44), and others have
adhered to the view that chemotropism is important, but more
recent work, such as that of Fulton (25), does not uphold the
theory.
[Vol. 1
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X
^
■>
^, f
294 ANNALS OF THE MISSOURI BOTANICAL GARDEN
\
Behrens (6) investigated some of the physiological relations
of saprophytes in comparison with parasites, using Mucor sto-
lonijcr, Penicillium sp., Botrytis cinerea, and Oidium { — Sclero-
titiia^) frudigenum. This author holds that Sclerotinia docs
not produce a cellulose-dissolving enzyme, and that the fungus
merely forces its way through the host tissue by a purely me-
chanical force, or that, in some cases, it sphts the middle lamella
but does not dissolve it. In the case of the other fungi mentioned
above he believes that an enzyme is secreted which dissolves
the middle lamella. The cause of the injury due to Sclerotinia,
he holds, is not that the cellulose walls or the pectin of the
middle lamella is dissolved, but that the turgor and the osmotic
relations of the penetrated cells are materially modified. Ac-
cording to this author some substance diffuses through the
walls and stimulates the fungus to bore through or between the
cell walls. He demonstrated in Botrytis and Pcnicilliujn, more-
over, a thermo-stable toxic body whicli disintegrated the host
cells, and believes that these fungi secrete a pectin-dissolving
enzyme which is different from that which acts upon cellulose.
Nordhausen (40), at about the same time, made similar studies
on Botrytis cinerea and comes to similar conclusions. He finds
that the enzyme does not cause a strong swelling of either the
middle lamella or the cellulose cell walls, the action in this respect
being more like that of de Bary's Sclerotinia. Smith (46) stud-
ied the parasitism of Botrytis cinerea, but in certain particulars
did not get the same results as de Bary and Ward. Like them
he finds that the parasite secretes some soluble substance that
penetrates and kills the living cells in advance of the fungous
filaments, but unlike Ward he could detect no swelling of the
cell wall. Smith beheves that this toxic substance is not an
enzyme, for boiling does not inactivate it, but thinks that it is
perhaps oxalic acid, since this substance is always present in the
cultures and amounts in some cases to as much as two per cent.
The analytical methods whereby the oxalic acid was determined,
unfortunately, are not given.
Schellenberg (43) investigated the action of several sapro-
phytic and parasitic fungi on hemicelluloses from a number of
1 Wehmer, C. Ber. d. dcut. bot. Ges. i6: 298-307. 1898; Saccardo, Syll. Fung.
4: 34. 1886.
1914]
COOLEY— SCLEROTTNIA CINEREA 295
different sources. He claims that these fungi act differently
toward different celluloses, dissolving some and having no effect
on others. The nature of the penetration and the action of
certain parasites on the host tissue were also studied. There
was no case in which Botrytis dissolved true cellulose, but it
readily dissolved the hemicellulose part of the cell; leaving the
cellulose intact. According to this author, therefore, the pene-
tration and dissolving action of such parasites as Botrytis vul-
garis is due to their ability to dissolve hemicelluloses. He
considers that the middle lamella is largely composed of hemi-
celluloses or closely allied substances. According to this view,
therefore, organisms that dissolve the middle lamella are essen-
tially hemiccllulose-dissolving forms. As a result of his studies
on Sclcrotinia fructigena and S. cinerea, Schellenberg finds a
different action on different fruits, but in no case does he report
a splitting of the cells along the line of the middle lamella, as
some previous investigators have reported. He believes that
there is a slight dissolving action on that part of the cell wall
which is in immediate contact with the fungous filament, but
that the rest of the cell wall remains intact. In the twigs also
he finds that the fungus dissolves the hemicellulose and leaves
the true cellulose unacted upon.
An extensive literature has developed concerning the enzymes
of importance in the nutrition of fungi, but since these invcsti-
gations either deal with saprophytes, or are only indirectly
concerned with the work to be reported in this paper, it will be
unnecessary to do more than mention some of the papers here.
Among the more important contributors may be mentioned
Ward (50, 52), who was the first to use pure cultures of a wood-
destroying fungus {Stereum)^ Biff en (9), who studied the biology
of Bulgaria polymorpha^ Bourquelot and Herissey (13), who in-
vestigated the enzymes in sporophores of Polyporus sulphureuSy
Czapek (18), who made his investigations with natural infections
of Merulius lacrymans and with other fungi, Kolmstamm (33),
who worked on some species of Merulius^ Buffer (14, 15), who in-
vestigated sporophores of Polyporus squainosus, Van Iterson (28),
who developed methods for isolating cellulose-dissolving bac-
teria and fungi, and Dox (19), who investigated the enzyme
action of species of Penicillium and Aspergillus. It is interest-
3
[Vol. 1
290 ANNALS OF THE MISSOURI BOTANICAL GARDEN
ing to note that although we have every reason to believe that
cytase is present in timber-decay organisms yet its presence has
been demonstrated only indirectly by cytological methods.
It is true, however, that many of the investigators mentioned
above who found no cytase used the sporophores in their
experiments and not the mycelium.
The status of the subject of the enzymes concerned in the
metabolism of parasitic fungi is given in Reed's recent publica-
tion (42), which concerns itself with the enzymes produced by
the parasitic fungus Glomcrella rufomaculans. This author has
proved that the parasite produces many of the enzymes that
had previously been reported for saprophytes, and by quan-
titative methods has demonstrated different enzymes acting
on the several classes of nutritive substances, such as carbo-
hydrates, glucosidcs, fats, and proteins. He did not, however,
investigate the cytolytic activity of the fungus but states that
the nature of the diseased host would indicate that cytase very
probably is not produced by this fungus. Peltier (41), as a re-
sult of his investigations with Botrytis Fuckdiana, finds that
the host cells are killed in advance of the fungous penetra-
tion, and that the parasite secretes a thermo-stable toxic sub-
stance, but, unlike Smith, finds no oxalic acid. The method
of testing for oxalic acid unfortunately is not given.
The action of bacteria on cellulose and other plant products
has been extensively studied by a number of investigators, but
for the purpose at hand it will suffice to cite some of the more
recent publications in which the earlier literature is reviewed.
The work of Jones (29, 30), which gives a good resum^ of the
early work on this subject, is reviewed below under the dis-
cussion of pectin.
McBeth and Scales (38) report that a number of bacteria and
fungi hydrolyze cellulose and claim that filamentous fungi play
a very important role in the destruction of cellulose in soils.
The cellulose-destroying fungi, according to these authors, act
differently toward different kinds of cellulose, but their experi-
ments do not seem to support this conclusion. Kellerman and
McBeth (32) have also contributed to our knowledge of the cyto-
lytic activity of fungi. Kellerman (31) has employed a method
I
1914]
COOLEY — SCLEROTINIA CINEREA 297
by which it is demonstrated that cytase diffuses in agar consid-
erably beyond the region of hyphal penetration, and that a
portion of the agar containing the enzyme dissolves cellulose in
a manner similar to that of the fungus itself.
The organism employed in my work was isolated from an
infected plum twig, at Madison, Wisconsin. The original cul-
tures were taken from a single colony in a Petri dish, this pro-
cedure giving reasonable assurance that I was working with
a single strain of the organism. Regarding the systematic
relations of this organism a word may not be out of place here,
since considerable confusion has arisen in the literature regarding
the specific name of the organism causing the brown rot of
stone fruits (27, 53, 37). Woronin (56) has made an important
contribution designed to establish the systematic position of the
two species Sclerotinia dnerea and S. frudigena. It has gener-
ally been held that S. frudigena causes the brown rot of stone
fruits in this country, while in Europe this fungus is found only
on pome fruits ; but Matheny (37) has recently given good evi-
dence tending to show that it is S. dnerea which causes the
brown rot of stone fruits both in this country and in Europe.
Experimental Studies
INFECTION
Some investigators, as, for instance, Zschokke (57), have held
that Sderotinia dnerea is unable to penetrate sound fruit, while
Smith (45), among others, has held that the fungus rapidly pene-
trates and infects sound and unwounded fruit (peaches) . Casual
observation in the field would seem to justify the former view,
for those fruits in contact with other fruits or twigs, and there-
fore liable to puncture or abrasion, are the ones that are usually
found infected; indeed, field observations and laboratory exper-
iments point to the conclusion that infection takes place much
more readily, especially with immature fruits, when the cuticle
is broken. One would, therefore, naturally raise the question
as to whether or not infection can take place when the cuticle is
unbroken, and if so under what conditions and in what stages
of the development of the fruit. During the summer of 1913
«
1
{Vol. 1
298 ANNALS OF THE MTSSOX^III BOTANICAL GARDEN
the writer performed a number of experiments which throw
more light on the question of the infection of the host.
Methods and Result s .—The methods employed were as fol-
lows: Plum twigs bearing leaves and fruit w^ere broken off and
brought into the laboratory, washed with a mercuric chloride
solution (1-1000) and in sterile water. They were then sus-
pended in sterile moist chambers prepared by placing moistened
absorbent cotton in the bottom of wide-moutlicd one-liter Erlen-
meycr flasks that had previously been plugged and sterilized.
Twigs having one or more green leaves were used in every case,
for in this way green plums hang on the twigs and remain alive
for some time. This method was especially applicable here, for
it enabled one to maintain absolutely sterile conditions in a
moist atmosphere and at the same time keep the host living
and in a normal condition. The results of these infection ex-
periments are given in table i.
Discussion of Re
dent that
cted as early as June 27, at which time they were
immature, in fact not more than half-grown. ' Infection did not
take place when a spore suspension was placed on very green
and immature plums unless the epidermis was broken or punc-
tured. There were, however, some instances where plums re-
mained healthy in the flask for two or three weeks and became
infected only after the lapse of time had brought about an
artificial maturity. On the other hand, plums that were ap-
proaching maturity, thougl
may be infected by aj
the
,ce which has not been punctured or injured
in any way. In this connection it should be mentioned that
infection was much more readily accomplished when two plums
were hanging so as to be in contact with each other than when
they were not touching. This, no doubt, was due to the fact
that a drop of water containing spores may be held between
the plums long enough for spore germination and infection to
take place. These results also indicate that infection takes
readih'
felt
: should be noted here that one can sometimes find plum
field onlv half-crown which are affected with the brown
1914]
COOLEY — SCLEROTINIA CINEIIEA
299
TABLE I
RESULTS OF INFECTION EXPERIMENTS WITH SCLEROTINIA CINEREA
Date
Fruit
Inoculating
material
Treatment
of surface
Method of
inoculation
Results
June 27
Green
plums
' Spore
suspension
Cuticle killed
by steam
Surface
" application
++=^
June 27
Green
plums
]
Spores
Skin punctured
with needle
Needle
puncture
+
July 2
Green
plums
1
Spores
1
Skin punctured
with needle
Needle
puncture
: + +
July 8
Green
plums
1
Spores
Skin punctured
with needle
1
Needle
puncture
1
*
++
1
July 8
1
Green
plums
1
Spore
suspension
Untreated
1
Surface
apphcation
1
\
July S
1
Sour
cherries
^ Skin punctured
Spores . , f „
^ with needle
Needle
puncture
+
July 23
G reen
plums
Spores
Skin punctured
wiih needle
Needle
puncture
++
Julv 23
; Green
plums
Spore
suspension
1
Untreated
1
1
Surface
application
July 23
Green
])lums
1
Spore
suspension
1
Skin cut
Surface
application
++
1
July 23
1
1
1
1
Ripe
plums
Mycelium
1
1
Untreated
Surface
application
++
1
1
July 23
1
1
1
1
Green
plums
Mycelium
Untreated
Surface
application
++
July 30
Green
plums
Mycelium
Untreated
Surface
application
++
Aug. 5
Green
plums
Spore
suspension
Untreated
Surface
application
+
1
Aug. 13
Xearly
ripe
plums
Spore
suspension
Untreated
Surface
application
4
++
^
Aug. 13
Ripe
plums
Spore
suspension
Untreated
Surface
application
+ +
* + + indicales that practically every inoculated fruit became infected.
+indicates that only a portion of the inoculated fruits became infected.
— indicates that none of the inoculated fruits became infected.
]
300 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
fungus, but so far as the writer's observation indicates, infection
in these cases takes place through the twig, or, in some cases,
through another pkim with which it is in contact and which in
turn is infected through the twig. Nevertheless, field observa-
tions also verify the laboratory work in that plums (especially
'certain varieties, such as Wood) when approaching maturity
may be infected in the field without being in contact with other
fruits and without having any visible punctures or wounds in
the skin. All these experiments and observations point to the
conclusion that penetration of the cuticle is a very important
factor in the infection of fruits, especially immature fruits; that
infection of very green fruits without punctures is rare; and, on
the other hand, that maturing fruits without punctures may be
readily infected both by spores and by a mycelial felt in the
field and in the laboratorv.
PENETRATION
The nature of penetration and the course of the hyphse of
parasitic fungi in piercing host tissue is an interesting and im-
portant question in connection with a study of the nature of
parasitic action. In the case of the brown-rot fungus growing
on the plum it is of importance to know whether or not the
hyplia3 merely follow the middle lamella; or whether they enter
the cells wherever they come in contact with them. Previous
investigators differ very widely in their opinions as to the nature
and course of the penetration of the fungus in question, a condi-
tion which is perhaps partly explained by the fact that different
hosts were employed in the various investigations. Further-
more, it appears that the methods employed in some of the
researches were not of such a character as to readily yield com-
plete information concerning all the facts in the case.
In my own work a study of the penetration of the host tissue
by the fungus was made by examining a number of sections of
infected tissue in which the disease had reached various stages
of development, and comparing them with sections of healthv
same
method
emnloved
Methods and ResuUs.Smsll pieces of fruit composed of
diseased and sound tissue were cut from plums inoculated with
t
i
i
)
1914]
COOLEY — SCLEROTINIA CINEREA 301
a pure culture of the fungus. These segments were immersed
in 70 per cent alcohol just long enough to partially kill the
fungous filaments and the host cells, yet not long enough to
discolor the sound tissue or to modify or change the color of the
diseased tissue in any way. From this material razor sections,
containing both diseased and healthy tissue, were made, stained
for a short time in eosin, and then partially desta'ned with
alcohol. If the pieces of plum had not remained in the alcohol
for a sufficient length of time, the razor sections were immersed
in 70 or 95 per cent alcohol before staining. By employing
this method it is possible to stain the fungous filaments deeply,
while the host tissue remains unaffected. Indeed, this method
permits of a rather sharp color differentiation between the
healthy and the diseased tissue, the latter being blackened by
the disease. This method, though quite applicable for the pur-
pose at hand, was primarily developed for another purpose,
which will be discussed below.
Since every fungous filament is very sharply differentiated,
one may readily study the course of the hyphse with reference
to the host cells. By staining, sectioning, and examining dis-
eased material taken from the margin of the infected area, one
finds the fungous hyphse penetrating the cells at any point of
contact; indeed, after examining a number of specimens by the
method reported above, the writer finds no indications that the
fungous hyphae follow the middle lamellae, as has been reported
by other investigators (57, 6) for pears and other fruits.
The above method also enables one to contrast the cell walls
of infected and penetrated cells with those of normal tissue.
It is entirely possible that the fungous filaments, on coming in
contact with a cell wall, secrete just enough enzyme to dissolve
their way through the cell walls, leaving the walls of the host
cells surrounding the hyphae entirely normal, i. e., without
swelling or disorganization.
Another and somewhat different experiment was performed
to get additional evidence on this point. From sound plums
which had previously been rendered sterile by washing in bi-
chloride of mercury solution (1-1000) and sterile distilled water,
free-hand sections were cut with a razor sterilized in 50 per cent
alcohol. The sections were arranged in hanging drop cultures
V
[Vol. 1
302 ANNALS OF THE MISSOURI EOTAXICAL GARDEN
and each inoculated with a drop of a very dilute spore suspen-
sion containing two or three spores per drop. The progress
of the fungus and the condition of the host cells were noted
from day to day but no visible disintegration of the cell walls
could be observed, nor did the fungus show any particular
affinity for the middle lamellse.
Conclusions. — We would conclude, therefore, as a result of
direct observation on the host tissue, that the fungus penetrates
the host very readily and rapidly, that it does not necessarily
follow the middle lamella? in the plum and the peach, and that
there is no visible general disintegrating action on the middle
lamella,' or on the cell walls of the living host.
ACTION OF THE FUNGUS ON THE LIVING HOST CELLS
A significant fact in the metabolism of the brown-rot fungus
is that it induces such an exceedingly rapid decay in the infected
fruits. This rapid decay might be connected both with a
rapid growth of the fungus and with a pronounced power which
the organism possesses of breaking down and changing the
constituents of the host. Moreover, several representatives of
the genus Sclcrotinia have been reported to have the power of
secreting an enzyme or some other substance which kills the
host cells in advance of penetration. Were this the case, it
would be expected that rapid decay would accompany the action
of the parasite. Is this view applicable to the action of Sclcro-
tinia cincrea? The investigators who have made a study of
this organism differ very widely in their views regarding the
effect which it has on the host tissues, and it seemed desirable,
therefore, to determine the relation of hyphal penetration to
the death of the cells.
Methods and Results. — In order to fix the material for this
study, it was found satisfactory to proceed as follows: Small
pieces of the host tissue were taken from the mai-gin of the dis-
eased area and placed in 95 per cent alcohol for a short time.
Free-hand sections were made of this material so as to include
both diseased and healthy cells, and the sections stained for a
short time in eosin and subsequently decolorized in part with
alcohol, if necessary to give the desired contrast. By this
i
f
1914]
COOLEY — SCLEROTINIA CINEREA 303
method the fungus may be distinctly differentiated from the
host tissue, the kiUing and staining agents having little or no
effect on the host cells. There is a more or less sharply differ-
entiated line of demarcation between the Injured and the sound
cells, as indicated by the darker color of the former. The effect
of the fungus is readily discerned by the blackening of the host
tissue, this being especially noticeable in green plums. The
discolored and poisoned cells are not at first plasmolyzed, and
it is to be noted here that discoloration rather than plasmolysis
should be taken as the index of the toxic action of this fungus
on its host. It should perhaps be mentioned here, too, that the
blackened cells shade off somewhat gradually into the hyaline
healthy ones, and that, therefore, there is not always a sharp
line of demarcation between the diseased and the healthy cells.
However, in spite of these difficulties, I was convinced, after
having examined a large number of sections of diseased and
healthy tissue, that there is no positive evidence that the host
cells are discolored, and therefore injured and poisoned, in
advance of actual penetration by the fungus.
The indirect method employed to determine the same point
consisted in applying to sound fruits an extract from decayed
plums. Fruits were disinfected with mercuric chloride solution,
washed in sterile distilled water, and inoculated with Sclerotinia
cinerca. When the plums had become thoroughly decayed
the juice was extracted and filtered under sterile conditions
through a Chamberlain filter. The juice thus obtained was
incubated for one week at a temperature of 22-25° C, and also
tested on nutrient agar plates, and found to be sterile by both
methods. From sound plums, which had been disinfected in
the usual manner, a cone-shaped plug was cut out and the
resulting cavity filled with this sterile extract, — the controls
being prepared in a similar manner, using sterile water instead
of the plum extract. The results were negative, that is, the
controls were not unlike those treated with the extract from
decayed plums.
The same experiment was repeated
by
using thin razor sections of both green and ripe plums, the
sections being made under sterile conditions as before, and ob-
served in a hanging drop of sterile juice from decayed plums.
1
[Vol. 1
304 ANNALS OF THE MISSOURI BOTANICAL GARDEN
By means of this method one could readily observe any changes
that might take place in the cells and make accurate compari-
sons with controls. Frequent observations were made, and
throughout this experiment, which continued for several days,
one could not distinguish between the appearance of those
sections in a drop of sterile water and those in the sterile extract
from decayed plums. It is possible and perhaps probable that
this fluid, being merely the juice of the fruit, was too dilute to
be effective, but the experiment was made because of the
possibility of positive evidence.
Discussion of Results. — The initial stage in the injury caused
by this fungus is shown by discoloration only and not by plas-
molysis, and therefore one cannot draw conclusions with ab-
solute certainty as to the poisoning effect of the extract on
the cells of a cut surface, for the latter turn brown as soon as
exposed to the air, just as when infected with the organism.
It was comparatively easy, however, to observe that the extract
had no effect on the cell walls, for no difference could be observed
between the cell walls of the tissue thus treated and those of
the control specimens. Even where the sections were left in
the extract for several days neither swelling nor disorganization
of the cell walls or middle lamelhc was noted. When sections
of plum tissue were inoculated with one or more spores of the
brown-rot fungus no cell-wall disintegration resulting from the
growth of the fungus could be observed. A comparative study
of sections of tissue, respectively exposed and not exposed to
the action of the extract from decayed fruit, showed that no dif-
ference could be detected between the two, and that, therefore,
no enzyme with a perceptible cytolytic action exists under these
conditions. It has been held by some, notably by Behrens (6),
that the injury to the host cell is largely physical in that the
fungus penetrates at such a prodigious rate that the fluids of the
host cell are allowed to escape with loss of turgor to the latter;
furthermore, that the osmotic equilibrium is soon destroyed,
asmolvsis and death
obable
tissue.
of the rapid injury to the host can be explained on purely
ical grounds, but this may not be the only factor involved,
ugh we do not now know what chemical activity of the
s cells may be concerned in the rapid killing of the host
1914]
COOLEY — ^SCLEROTINIA CIXEREA 305
ACTION OF THE FUNGUS ON CELLULOSE
A number of investigators have regarded cellulose dissolution
as a very important factor in the parasitism of many fungi;
indeed, some of the earlier workers seemed to consider this the
prime factor involved. While it is a well known fact that there
are many fungi, especially saprophytes, which hydrolyze, or dis-
solve, certain celluloses, research extending over a wide field
has revealed the nature of parasitism to be a very complex one
in which other factors are as important as the dissolution of cel-
lulose and the cell wall.
It has been the writer's purpose to study from two different
points of view the action of the brown-rot organism on celluloses,
(1) by observing the action of the fungus on pure cellulose iso-
lated from the host tissue, and (2) by studying microscopically
its action on the host cell walls themselves. In the former
study cellulose agar was used, the cellulose being isolated from
plums by the methods discussed below\
Methods and Results. — In the above mentioned study of the
action of the fungus on pure cellulose, a variety of reagents,
media, and methods for the preparation of cellulose were em-
ployed, a brief account of which follows. Schweizer's reagent
was prepared by adding a slight excess (40 grams to the liter)
of copper carbonate to dilute ammonium hydroxide solution
composed of three parts of water to ten parts of ammonium
hydroxide (sp. gr. 0.90). The copper solution was then shaken
vigorously, allowed to stand over night, and the supernatant
solution siphoned off. This is the procedure employed by Mc-
Beth and Scales (38).
Paper cellulose from filter paper was prepared according to
the method given by McBeth and Scales (38) by dissolving 15
grams of sheet filter paper in Schweizer's reagent, diluting about
ten times with water, and precipitating the cellulose with a
solution of one part of hydrochloric acid to five parts of water.
This mixture was then further diluted to 15 or 20 liters, the
supernatant liquid siphoned off, and the residue washed re-
peatedly with water until the precipitated cellulose was free
from both copper and chlorine. After standing quietly for
several days the clear liquid was siphoned off and the precipitate
used for the preparation of cellulose agar.
[Vol. 1
306 ANNALS OF THE MlSSOriU BOTANICAL GARDEN
Cellulose agar was made by adding about one per cent (esti-
mated by the weight of the paper before treating with Schwei-
zer^s reagent) of precipitated x>aper cellulose, prepared as stated
above, to a mineral nutrient solution^ the complete medium
having the following composition :
Cellulose suspension 500 cc.
Agar 10 grams.
Alonopotassium phosphate, 1 gram
Magnesium sulphate, 1 graui
Sodium chloride, 1 gram
Animoniuin sulphate, 1 gram
Calcium carbonate, 2 grams
Tap water, 1000 cc.
The insoluble precipitate appearing in the mineral nutrient
solution was filtered off before the cellulose suspension and agar
were added. Good results were also obtained by using 0.5
gram of calcium nitrate instead of 2 grams of calcium carbonate,
in which case filtering is unnecessary. The mineral nutrient
solution having the composition tabulated above will be referred
500 cc.
to as nutrient "A."
Another nutrient solution very low in organic matter was
also employed in the cellulose agar, but with rather unsatis-
factory results. This solution, which will be referred to as
nutrient "B,'' is that employed by Reed (42), and is made up
as follows, the only organic material present being the small
amount of sodium citrate :
fframs
Ammonium nitrate 10
Dipotassium phosphate " 5 grams
Magnesium sulpliate 1 gram
Sodium eitratr
1 ffram
Tap water 1000
cc.
In making the cellulose agar this nutrient solution was used in
exactly the same way as nutrient "A."
Since previous investigators have held that the celluloses from
various sources differ in their resistance to hydrolyzing enzymes,
an attem.pt was made in this investigation to prepare a cellulose
from a natural host — plums — of the parasite. In order to secure
a cellulose that is modified as little as possible in the process
1914J
COOLEY — SCLEROTINIA CINEIIEA 307
of isolation three different methods were employed in preparing
from
ducts being designated
for convenience in reference, respectively as soda cellulose,
washed cellulose, and potassium chlorate cellulose.
In the preparation of soda cellulose ripe plums were squeezed
through cheese cloth and the pulp was washed thoroughly with
water. The pulp was then treated with an 8 per cent solution
of sodium hydroxide and heated in the autoclave at- ten pounds
pressure. After thoroughly washing the pulp with water the
heating with alkaU was repeated and the product given final
washings until free from alkali.
The second method of isolating cellulose — washed cellulose —
consisted in washing the fruit pulp with water until free from
substances soluble in cold water. Water was then added and
the mixture heated in the autoclave at 15 pounds pressure,
and washed. The operation was repeated as long as any water-
oluble substances could be detected. This method, of course,
;ives an impure cellulose, yet the product is one that is free from
i^ater-soluble substances.
The third method consisted in oxidizing, dissolving, and wash-
the
chlor
cellulose— was obtained. Pulp, secured from ripe plums in
the manner stated above, was washed with cold water until
the wash water was free from solutes, and then treated with a
cold solution composed of 30 grams of potassium chlorate dis-
solved in 520 cc. of cold nitric acid (sp. gr. 1.1). This mixture
was kept in the ice box for about three weeks, at the end of
which time the pulp was entirely white. This method ^ is said
to yield a product that differs only very slightly from the original
cellulose.
obtained by these various methods
lowed to dry, for it is possible that drying changes the nature
of cellulose so that it is more resistant to the action of cytolytic
enzymes. A part of the cellulose obtained by each of the pre-
ceding methods was treated with Schweizer's reagent and pre-
cipitated with hydrochloric acid and washed as stated above
under the preparation of filter-paper cellulose. These three
ffillnlose nreoarations thus treated with Schweizer's reagent, as
1
Fowler, G. J. Bacterial and enzymatic chemistry. 159. 1911.
308 ANNALS OF THE MISSOrHI BOTANICAL GAKDEN
[Vol. 1
well as the three corresponding untreated portions, were used
in the preparation of cellulose agars, according to the method
given above. The media were placed in test tubes of very small
mm.) diameter, and sterihzed. The tubes of melted agar
were then cooled rapidly in cold water in order to bring about
the hardening of the agar before the cellulose had had time to
settle to the bottom of the tubes.
Tubes of the various cellulose agars were inoculated with
Sderotinia cinerea and others with a species of Penicillium,
which will be designated as P. expansum\ isolated from decay-
ing peaches and apples. Since these two fungi, viz., Sclerotinia
cinerea and Penicillium expansum, act very differently toward
the host, a word contrasting their action may not be out of
place here. As a result of inoculating apples, peaches, or pears
with a pure culture of Sclerotinia the host tissues are promptly
killed, while the fruits remain practically as firm after complete
decay as before inoculation. On the other hand, the fruits inocu-
lated with the Penicillium become very soft and watery, develop-
ing a pustule or sunken area where the infection took place.
One may assume, therefore, that the Sclerotinia does not materi-
ally affect the celluloses and pectic substances that make for the
firmness of the fruit, while, on the other hand, Penicillium does
alTect these substances, causing the fruit to lose its firm consis-
tency. Since these two fungi show such entirely different and
opposing characteristics as regards their effect on the same
host, it is interesting to compare their action in pure cultures on
cellulose and pectin-hke substances. Such a comparative study
was made, the results of which are given in table ii.
Discussion of Results.— The results given in table ii indicate
both Sclerotinia cinerea and
ited
f
drolytic action when
cellulose isolated from the plum, there being very slight action
with both fungi on the soda cellulose and also on the potassium
chlorate cellulose and no action on the washed plum cellulose.
On the other hand, both fungi very readily dissolve filter-paper
' A culture of this organism was sent to Dr. Chas. Thorn, who very kindly examined
it and gave as liis opinion that it was P. expan.wm, or perhaps a strain of that species.
The organism in question, when grown on the media employed by Thorn, showed
characters very similar to those of P. expansnm, as given by Thorn (48).
1914]
COOLEY — SCLEROTINIA CINEREA
309
TABLE II
ACTION OF SCLEROTINIA CINEREA AND PENICILLIUM EXPANSUM ON CELLULOSE
■
1
1
Nutrient
Sclerotinia cinerea '
1
Penicilllumcxjjansura
Type of cellulose used
solution
added
1
Growth
Cellulose
hydrolysis
1
1
Growth
CelTulose
hydrolysis
Soda cellulose
A
+ +t
1
+ +
+
Soda cellulose
B
+ +
Potassium chlorate cellulose
A
+ +
1
+
+ +
+
Potassium chlorate cellulose
;
B
+ +
1
+
1
1
Washed ligno-ccllulose*
A
+
1
\\ ashed ligno-cellulose*
B
+
'
1
\\'ashed cellulose
A
—
+
Soda cellulose (Schweizer's)
B
+
F
r
1
+
Soda cellulose (Schweizer's)
A
1
T
+ +
+
1
1
1
\\'ashcd cellulose (Schweizer's)
1
A
+
X
1
1
Soda cellulose
1
Peach
juice
1
^_ ^_-
1
+ + +
1
+ + +
1
1
Filter paper strii)s
1
Peach
juice
1
+ + +
+ + +
—
Filter pai)cr strips
A
+ +
™
+ +
^
Filter paper strips
B
+
+ +
II ■ . — - '^
Filter paper strips
0.5%
glucose
solution
r
+ + +
+ + +
.....
Filter-paper cellulose
A
+ +
+ + +
+ + + + +
iigno
plum, i. e., that part of the pulp which did not go through the cheese cloth.
fGrowth and cellulose hydrolysis are indicated by +, the relative intensities of
growth and degrees of hydrolysis being indicated by one or more + marks. Absence
of growth and absence of hydrolysis are indicated by — .
310 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
cellulose, and, strange to say, Sclerotinia is just as active in this
respect as Pcnicillium. In many cases the growth was as good
on the plum cellulose as on the filter-paper cellulose, yet the
hydrolytic action of the fungi was very much weaker on the
former medium. No cellulose hydrolysis occurred where peach
juice or some soluble carbohydrate, such as glucose, was added.
It seemed probable at first that a very small amount of glucose,
or peach juice, or sodium citrate would give the fungus a vigor-
ous start and thus accelerate its cyto-hydrolytic activity, but
the quantities of these substances employed was sufficient to
exert a protective influence, there being a vigorous growth but
no apparent cellulose hydrolysis.
The fact that these fungi do not dissolve cellulose, derived
either from the host or from paper, when other organic nutri-
d, verifies the writer's observation that Sdei
d
of th
Furthermore, the fact that the fungus dissolves paper cellulose
very readily when it is the only carbohydrate supplied, leads
one to conclude that the action of the fungus on paper cellu-
lose in a nutrient solution low in carbohydrates is not neces-
sarily a good criterion for judging the behavior of the fungus
in the host tissue. In the host tissue there may be a form of
cellulose different from that of paper, and it is furthermore
very evident that there is present in the fruit an abundance of
organic material evidently operating in a protective manner.
The fungus fails to produce cytolytic enzymes when grown on
plum or paper cellulose to which peach juice or even a very little
sugar has been added, but acts vigorously on paper cellulose
to which no organic nutrient has been added. It is rather
peculiar that both fungi act much more readily on paper cellu-
lose than on cellulose isolated from the fruits which are natural
hosts for these organisms.
Sclei
transferred
■lent medium poor in soluble carbohydrate
d no aerial mycelium being produced. At
ated
observe that the fungous
' layer of the agar, and
5, in case the fungus is g
may be observed
I
1914]
COOLEY — SCLEROTINIA CINEREA 311
in the agar just below the fungous filaments, thus indicating
that the cellulose is being hydrolyzed. With increasing age of
the fungus, this clear and almost transparent area gradually
enlarges downward, although the fungus shows little or no cor-
responding penetration. At the expiration of three weeks or a
month, there is a very distinct, clear, and nearly transparent
zone in the medium below the region occupied by the fungous
mycelium. Since one could see very distinctly how far the
fungous filaments had penetrated into the substrate, it was
very evident that the cyto-hydrolytic enzyme had diffused
beyond the limits of the mycelium.
The method employed in this investigation for the demonstra-
tion of cellulase was the same as that used by Kellerman in his
recent work (31) and was utilized to demonstrate the fact that
the cyto-hydrolytic enzyme secreted by this fungus penetrates
the substrate considerably beyond the limits of the filaments
themselves. Tubes containing cellulose agar, in which the
fungus had been growing for four weeks, were disinfected exter-
nally by washing with a bichloride of mercury solution, and cut
off at a point about 12 mm. below the clear portion of the me-
dium
then flamed and pushed
tube with a glass rod until the agar was partially shoved out
of the cut end of the tube. The clear portion of the agar was
then cut into disks about 12 mm. in thickness, which were laid
on plates poured with nutrient cellulose agar, great care, of
course, being exercised throughout the operation to maintain
aseptic conditions. The plates so prepared were then placed
in an incubator at 25°C, where they remained for two weeks,
at the expiration of which time the cellulose was very distinctly
hydrolyzed in a ring about the sterile shces of agar. Micro-
scopic examination confirmed the macroscopic observation that
these agar disks were free from any infection.
As might be expected, the activity of the secretion
enzyme cellulase is influenced by temperature, a fact which is
well illustrated by the following experiment: Tubes containing
cellulose agar inoculated with the brown-rot fungus were kept
at temperatures of 10-12, 16-20, and 24-26°C. respectively, and
at the end of twelve days the following results were noted: In
the cultures maintained at 10-12''C. no apparent growth or
4
of the
I
[Vol. 1
312 ANNALS OF THE MISSOURI BOTANICAL GARDEN
hydrolysis had taken place; those kept at 16-20°C. showed a
good growth but no visible cellulose hydrolysis; and in those
maintained at 24-2G°C. there was about the same extent of
growth as in the preceding series but accompanied by a very
evident cellulose hydrolysis, a distinctly clear zone of dissolved
cellulose surrounding the region occupied by the fungous myce-
lium. It is therefore evident that even with approximately the
same amount of growth cellulose hydrolysis is much more rapid
at the higher temperature.
An effort was made to determine whether or not it is possible
to ''train up" more active cyto-hydrolytic strains of the Sclero-
tinia and Penicillium. in question. On the one hand, these fungi
were grown for several successive generations on peach-juice
agar — a medium in which the organisms show no cytolytic ac-
tivity. On the other hand, these fungi were cultivated for sev-
eral successive generations on paper-cellulose agar — a medium
which is low in soluble carbohydrates, and one in which the
fungi exhibit considerable cytolytic activity. Tubes of paper-
cellulose agar were then inoculated with the fungi grown in
these two ways and careful observations were made to detect
any differences in cyto-hydrolytic activity. No differences
developed, however, from which it would appear that the source
of cultures of Sclerotinia or of Penicillium does not materially
affect the cellulose-dissolving capacity of these organisms, i. e.,
-r
each fungus shows the same cellulose-hydrolyzing power wheth-
er the organism was cytolytically active during the immedi-
ately preceding generations or not.
EFFECT OF THE FUNGUS ON PECTIC SUBSTANCES
The power of organisms to change pcctic substances has.
been considered an important factor in the disintegration and
softening of host tissue by certain plant parasites- Before enter-
ing into a discussion of the experimental phases of this subject,
it will perhaps be well to give some idea of the present status of
this question, as well as a very brief resum6 of the extensive
Hterature which has accumulated about it.
Fremey (23, 24), in 1840, was the first to report an enzyme act-
ing on pectic substances. This enzyme, which he isolated and
called pectase, induced the coagulation of pectin, Fremey attrib-
I
i
1914]
COOLEY — SCLEROTINIA CINEREA 313
uting this action of the enzyme to the presence of calcium salts.
It is of interest to note that pectase was one of the first plant
enzymes to be described. Bertrand and Mallevre (7, 8) con-
cluded that pectose and pectase are almost universally present
in green plants, being especially abundant in the leaves. These
authors showed that acidity is an important factor in the inhi-
bition of coagulation of pectic bodies by pectase, and also that
of
barium, calcium, or strontium is necessarv for the
. Mangin (35, 36), by microscopic tests, has thrown much
Hght on the nature of the middle lamella and holds that pectose
is very pronounced in the cell walls of young tissue. In the
older cell walls, on the other hand, this author beheves that
calcium pectate predominates in the middle lamella, considering
that the latter is largely if not entirely composed of this sub-
stance and that it frequently collects on the surface of the cell
walls adjoining intercellular spaces. Bourquelot (11), and
Bourquelot and H^rissey (12) secured a thermo-labile enzyme
from barley malt extract which acted upon a solution of pectin
(taken from the gentian root), changing the latter in such a
way that it was no longer coagulated by pectase. The action
of this enzyme, which they called pectinase, was thought by
them to be that of converting the pectin into reducing sugar.
They also designated as pectinase an enzyme which dissolves the
pectic coagulum (the latter has been supposed to be calcium
pectate). A good resum^ of the status of the chemistry of
pectic substances is given by Bigelow and others (10).
A number of investigators have reported upon the action
of bacteria on plant cells, including the effects of the organisms
on the middle lamella. Winogradsky (55), Behrens (5), and
others attributed the changes taking place in the flax plant dur-
ing retting to the dissolving action which the bacteria exert on
the middle lamella. It will be unnecessary to review here any
more of the earlier work which has been done along this line,
since it has been so thoroughly discussed in the comprehensive
pubhcations by Jones (29), and Jones, Harding, and Morse
(30) on the soft rot of vegetables. These authors studied the
effect of the soft-rot bacillus {Bacillus carotovorus) on the host
and find that the organism is identical with what has been
>
I
[Vol. 1
314 ANNALS OF THE MISSOLRI BOTANICAL GARDEN
designated as B. oleracece Harrison, and B. omnivorous Van Hall,
and that it may possibly be identical also with Potter's B.
destructans. By many tests Jones has shown that this organism
secretes an enzyme which causes the disintegration of the host
cells by dissolving the middle lamella, which, according to the
majority of investigators, is composed of salts of pectic acid.
This author has further isolated from pure cultures of the or-
ganism an extra-cellular enzyme, which he designated pcctinase,
that destroys the middle lamella of the cells just as does the
growing organism. Jones, therefore, considers this enzyme
responsible for the disintegrating action of the bacillus.
In my own work I shaU adopt the nomenclature used by Jones
('29. 30) and Euler (21), namely, employing pectinasc as the term
to designate the enzyme inducing coagulation of a pectin solu-
tion and also the hydrolysis of calcium pcctate, or pectinate.
Methods.— In order to determine the effect of the fungus on
the middle lamella I have used two methods, (1) a microscopic
study of the effect of the fungus on the host cells, and (2) a
study of the effect of the organism on the substances (isolated
from the host) which are commonly reported to be constituents
of the middle lamella. The first method has been discussed
above and may be dismissed here by stating that it yielded no
positive evidence that the fungus dissolves the middle lamella.
By the second method the problem was studied by isolating pec-
tin from the host and studying the effect of the fungus on it
and also on its salts, as, for instance, calcium pectinate.
Pectin was isolated from plums by the following method:
Thoroughly ripe fruits were steamed— no water being added, the
juice filtered off and treated with Almen's reagent' (to precipi-
tate the protein) and with a very dilute solution of oxalic acid
(to precipitate the calcium). It was found that under these
conditions neither a calcium nor a protein precipitate was thrown
down either by Almen's reagent or the oxalic acid, and this pro-
cedure, therefore, was deemed unnecessary and was abandoned.
The plum juice was carefully filtered through a Buchner filter
lAbdorhaldcn, E. Handbuch d. biochcm. Arbeitsmethoden 2 : 391-92. 1910.
Almen's tannic acid solution is made by treating 4 grams of tannic acid with 8 cc.
of a 25 per cent solution of acetic acid, and making up to 190 cc. with 40 or 50 per
cent alcohol.
4
)
\
K
1914]
COOLEY — SCLEROTINIA CINEREA 315
and the filtrate treated with 95 per cent alcohol until a floccu-
lent coagulum of pectin was produced. This pectin was sepa-
rated by means of a Buchner filter, redissolved in water, reprecip-
itated with alcohol, again separated by means of a Buchner
funnel, and finally dried at a temperature slightly higher than
room temperature,— the reprecipitation being for the purpose
of purification. It should be noted here that the plums were
sufficiently acid to make the addition of hydrochloric acid to
the alcohol unnecessary.
Experiments with pectin and peclinase. — From the pectin
isolated by the above method a saturated aqueous solution was
prepared — some of the mineral nutrient solution^ minus calcium
being added, and the resulting solution rendered sterile by frac-
tional sterilization. Test-tubes of this pectin solution were
inoculated with Sclerotinia cinerea and Penicillium expansum
with the result that both organisms produced a rather vigorous
growth of myceUum and a few spores. At the expiration of
one week the inoculated tubes showed a shght clear area just
below the fungous felt due to the coagulation and settling out
of the pectin in that part of the solution. The coagulation was
at this time somewhat more pronounced in the Penicillium cul-
tures than in those of Sclerotinia, yet very noticeable in both
cases, beginning directly below the fungous felt and progressing
toward the bottom of the tube. After two weeks the greater
part of the pectin solution was coagulated, the flocculent coagu-
lum, or precipitate, being very different from the precipitate
produced in a pectin solution by a calcium salt. It should be
emphasized here that every precaution was taken to maintain
a calcium-free solution, and when it is considered that the addi-
tion of calcium develops a reaction very different from that
produced by the enzyme, and, furthermore, that the check gave
no coagulation whatever, not even when allowed to stand a
month or more, the conclusion would seem to be warranted that
calcium is not necessary for the production of a gel by pectinase.
Both Sclerotinia and Penicillium, therefore, produced a coag-
ulum in an aqueous solution of pectin, while no such results
were obtained in the controls, thus justifying the conclusion
'Nutrient solution employed was the same as mineral nutrient solution A used in
preparing cellulose agar, but without the calcium.
r
[Vol. 1
316
ANNALS OF THE MISSOURI BOTANICAL GARDEN
f
that these two fungi are capable of producing pectinase. The
cultures were kept at a temperature of 18-20°C.
Experiments with calcium pectinate.— Cdlcmm pectinate was
prepared by treating a water solution of pectin with freshly-
made lime water (care being exercised to avoid an excess of lime),
the product thus obtained being filtered off and thoroughly
washed until it was no longer alkaUne. The calcium pectinate
thus prepared was used in making a pectinate agar in a manner
similar to that employed in the preparation of cellulose agar,
the same mineral nutrient solution (nutrient A) being used and
the whole rendered sterile by fractional sterilization. After
the last heating, care was taken to distribute the pectinate,
which quickly settles to the bottom of the tubes, uniformly
throughout the agar by stirring the medium with a sterile glass
rod. These tubes were then inoculated with Sclerotinia and
with Peniciltium, the object being to compare the action
toward pectic substances of two fungi that have entirely dif-
ferent effects on the host
s
)
the former prod
ing effects, while the latter causes a very rapid softening and
disorganization of the host tissue.
The inoculated tubes of pectinate agar prepared by the above
method were kept at a temperature of 22-24°C. Contrary
to expectations, there was very Httle growth when no soluble
bohydrate was
the calcium
per cent
plied, and, furthermore, no dissolving
Dectinate. On the other hand, when 0.5
added, both fungi produced a vigorous
but neither one gave any indication of pectinate hyd
sis, or dissolution. Here
drolysis
the two fungi, Sclerotinia and Pcnicillium, behave alike
This
is not in accordance with the observed bch
organisms toward the host tissue.
of
ACID RELATIONS OF THE FUNGUS
Some investigators have held that the content of tannin (47)
and of maUc and other acids of the host determines whether or
fungus can grow in the tissues and rot the fruit
In
accordance with this view a fungus may not so readily attack
green as ripe fruit, the former being sunnosed to exhibit a higher
J
H
^
1914]
COOLEY— SCLEROTINIA CINEREA 317
of these
The Question of the acid
relation of the host tissue is one of fundamental significance
and one that is worthy of considerable investigation; it is im-
portant to know to what extent acidity may be a Umiting factor
in parasitism.
A case in which a certain acid content is favorable for the
fungus is developed by Falck (22). He finds the acidity of the
substrate to be a conditioning factor for the growth of several
species of Merulius. In this connection the author observes that
Coniophora, in particular, acts to pave the way for Merulius in
that the former organism renders the nutrient substrate deci-
dedly acid, and thereby provides favorable conditions for the
germination of the spores and the subsequent growth of myce-
lium and fruit bodies of Merulius. In connection with the in-
vestigation of the plum disease here discussed it would be well
to know if the acidity of the fruit changes during the progress of
its growth, and if so in what direction. It is also essential to
know whether or not a change in the acidity of the host can
account for the fact that ripe fruit is more susceptible to the
disease than green fruit. Some experiments were planned,
therefore, to determine to what extent the acidity of the host
influences the attack of the parasite, and also to investigate
what effects, if any, the fungus has with respect to the acid
content of the host.
In order to determine the changes in acidity which take place
during the growth of the fruit (plums), several analyses for acid-
ity were made at intervals during the summer. The plums for
all of the analyses were taken from the same tree, a known
weight of pulp being ground up in a mortar and squeezed
through mushn. The acidity was reckoned in the number of
cc. of N/10 NaOH required to neutraUze one gram of plum
pulp. The results were as follows:
June 28, 1 gram plum pulp required 0.66 cc. N/10 NaOH for
neutralization,
Aug. 2, 1 gram plum pulp required 2.12 cc. N/10 NaOH for
neutralization,
Aug. 19, 1 gram plum pulp required 2.46 cc. N/10 NaOH for
neutralization,
318
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
the fruit being market ripe on August
In these tests my
for peaches, and
for some other fruit
th those obtained by Bigelow and G
(10)
th those of Thompson and Whittier (49)
3. The last mentioned investigators, how
rity.
found that the acidity of peaches decreases toward matu-
have been unable to secure data covering the acidity of
plums throughout the
The above results show that the acid content of plums in-
creases rather than diminishes toVard the maturity of the fruit.
The results of the experiments and field observations show that
mature and ripe fruit is much more susceptible than the green
and immature fi
The above facts, showing that as the fr
approaches maturity the acidity increases while the suscepti-
bihty to the disease also increases, indicate that there is no
close relationship between the low acid content of the host and
susceptibihty to the brown-rot fungus, and that we must look
to other factors to explain infection as observed in the field.
As pointed out, my experiments indicate that Denetration is a
TABLE III
REL.VTION OF THE GROWTH OF SCLEROTINI.\ CINEREA TO THE RE\CTIUN OF
THE MEDIUM
Medium
Cherry juice
Acidity
+2.3
Cherry juice
+1.5
Cherry juice
+ 1.0
Cherry juice
+0.15
Cherry juice
-0.15
Cherry juice
-o.r^o
Growtli after Growth after
8 days
t
IG days
+t
++
++
+++
++
+
++
++
Spore
I^roduction
+
+ +
+ +
+
♦Acidity IS given in cc. of N /lO xXaOII necessary to neutralize 1 cc. of the juice.
TThc + sign indicates a fairly good mycelial growth, or spore format ion, and the
- sign indicates that the growth was ju.'^t perceptible; indicates no growth, or no
spore formation.
1914]
t
1
^
*
COOLEY — SCLEROTINIA CINEREA 319
very important factor. It is possible that a study of the tannin
content' might yield some relation of interest.
A preUminary experiment was planned to determine the
acidity at which the optimum growth and spore production of
the fungus occurs. For this purpose the juice from ripe sour
cherries was used. The juice was squeezed out of the cherries
(no water being added) and a portion titrated to determine the
acidity. Then 50 cc. of this liquid were put into each of a
number of Erienmeyer flasks of 125 cc. capacity; some of the
flasks were left untreated, while others received various quanti-
ties of N/10 NaOH to bring each to the desired acidity or alka-
linity. The flasks were then sterilized and inoculated. The
results are given in table iii.
It is clear, therefore, that although the fungus eventually
grows on a medium as acid as the natural juice of sour cherries,
it grows more luxuriantly on a somewhat less acid medium. It
is a rather significant fact that on the media near the neutral
line the fungus at first shows no perceptible growth, but at the
expiration of two weeks has produced nearly as much mycelial
growth as on the acid medium. It is also of interest to note
that we find spore formation abundant on the very acid media
but entirely lacking on the alkaline media. This experiment
indicates that the fungus can adjust itself to a slight degree of
alkalinity.
OXALIC ACID PRODrCTION BY THE FUNGUS "
The first important reference to oxalic acid production by
fungi is in the publication by de Bary reviewed in a preceding
section. He reports that the older hypha^ of the fungus were
encrusted with crystals of oxalic acid, and he attributed some
of the poisonous action of the parasite to the production of this
substance; in fact, he mentions oxalic acid fermentation. Since
the appearance of de Bary's paper a limited number of investi-
^ Cook and Bassett and their associates (17) believe that there are enzymes in the
host plant which may act upon cell constituents and play the role of alexins. They
are of the opinion that tannin, as such, is not abundant in fruits, but that it may be
formed by the action of oxidizing enzymes upon certain phenols. Injuries produced
by parasitic fungi may accelerate the activity of the host in the production of tannin,
the latter perhaps being toxic to the growth of parasitic fungi.
[Vol. 1
!
f
320 ANNALS OF THE MISSOURI BOTANICAL GARDEN
r
gators have reported the presence of oxaHc acid resulting from
the growth of both fungi and bacteria, but unfortunately much
of this work is of little value, because methods of analysis are
not given. The detection of this acid by some methods is very
unsatisfactory.
A few years after de Bary's work, Wehmer (54) published an
extensive series of articles on this subject. He studied a number
of fungi (mostly saprophytic) with reference to oxalic acid excre-
tion, and of these he found Aspergillus to be the most active and
Pcnicillium next, and, therefore, he confined his studies to these
two fungi. Some of the factors concerned in the production of
oxalic acid or its salts, according to Wehmer, may be summed
up here: (1) A large yield of oxahc acid is not produced in the
presence of free organic or inorganic acids, not being found in
the medium when free acids exceeded 0.2-0.3 per cent, while,
on the other hand, it can be formed in the presence of as much
as 2-3 per cent of the salts of these acids. (2) The sources of
nitrogen are very important, for the amount of the oxalic acid
produced varies according to the kind and quantity of nitrog-
enous compounds supplied. (3) Abundant oxalic acid forma-
tion is favored by the addition of some basic phosphate, or at
least some compound with which the acid can combine to form
a soluble salt. (4) The effect of light or darkness on oxalic
acid formation is inappreciable. (5) Temperature is an influ-
encing factor in oxalate production, for the latter is inhibited by
a high temperature, the temperature for a maximum oxalate
production being, in fact, very near the minimum for the growth
of the organism.
Wehmer's analytical method consisted in precipitating out
the oxalic acid, or its soluble oxalate, as the calcium salt, which
was filtered off, dried to a constant weight, and weighed. Al-
though this method is perhaps as well suited for this purpose
as any other reported, it is open to criticism. A detailed dis-
cussion, however, will not be given here.
Wehmer holds that oxalic acid is a type of excretion, and that
it is in some way connected with respiration, that is, with CO 2
elimination. He considers that the variability in the amount of
oxalic acid produced is due to its use in the metabolism of the
fungus. Emmerling (20), in his contribution to this subject,
f
i
I
i
J
\
1914]
COOLEY — SCLEROTINIA CINEREA 321
emphasizes the influence of such nitrogenous substances as
proteins, amino acids, and amides in the nutrient. He finds
that Aspergillus niger when grown in non-amino acids, for
example, tartaric, lactic, etc., produces no oxalic acid, whereas
an abundant oxalic acid production results on such substances
as peptone or aspartic acid.
Smith (46) and Peltier (41) both conducted experiments to
determine whether or not oxalic acid is present in media in
which Botrytis has been growing. Peltier reported negative
results, but Smith found oxalic acid and thinks that the pois-
oning effect of the fungus is perhaps due to the presence of this
acid. Unfortunately, neither of these authors gives his methods
of analysis, and, with the exception of one incident in Smith's
publication, the quantity of oxalic acid found is not reported.
Peltier and others have been able to produce an injury with
oxahc acid similar to that produced by certain parasitic fungi,
such as Botrytis, yet this is not conclusive evidence that oxalic
acid is the toxic substance secreted by the organism.
The articles mentioned above constitute the chief publica-
tions that have to deal with the production of oxalic acid by
fungi. The publications on the production of oxalic acid by bac-
teria and other plants will not be reviewed here. Whether ox-
alic acid production is a phenomenon peculiar to certain genera
or to certain species of the fungi, whether it is purely the result
of external conditions, or whether it results primarily from cer-
tain constituents of the medium, has not been clearly demon-
strated. A series of experiments was planned in the hope of
throwing some light on its production in the fungus here studied.
The method of analysis employed was a modification of Weh-
mer's method of precipitating the oxalate with calcium chloride
and determining the amount of oxalate thus precipitated.
This method, however, is not well adapted to the purpose at
hand, especially when quantitative methods are used, and fruit
juice is employed for the medium on which to grow the fungus.
An attempt is being made to develop a method that will be
better suited to our purpose.
Culture media were prepared from peaches and plums by
filtering the juices of these fruits through a Hill pressure filter
under sterile conditions. The product thus obtained was
[Vol. 1
322 ANNALS OF THE MISSOURI BOTANICAL GARDEN
placed in flasks and incubated for a week and found to be sterile,
after which the flasks were inoculated with Sclerotinia cinerea.
At the expiration of thirty-seven days these cultures were ana-
lyzed and were found to contain the following amounts of oxalic
acid per 50 cc. of the respective juices:
Plum juice 0.0019 grams of oxalic acid,
Peach juice 0.0077 grams of oxalic acid,
Peach juice 0.0094 grams of oxalic acid,
Control No trace of oxalic acid.
Plum and peach juices that had been sterilized by heat; thereby
precipitating some of the contained proteinaceous material,
were also used as culture media, and here, too, every culture
containing the fungus gave a positive test for oxalic acid.
For investigating the production of oxalic acid by the fungus
in the unaltered fruit, lots of 500 grams each of peaches were
disinfected with bichloride of mercury solution, inoculated
respectively with Sclerotinia, Penicillium, and Aspergillus niger,
and kept under sterile conditions until the fruits were decayed,
or, in the case of the Fenicillium and AspergilluSy until partially
decayed. The decayed fruits were then digested with hy-
drochloric acid and analyzed for their oxalic acid content with
the following results:
Poach inoculated with Penicillium . . No trace of oxalic acid,
Peach inoculated with Aspergillus . . .No trace of oxalic acid,
Peach inoculated with Sclerotinia cinerea
t
0.0087 grams of oxalic acid.
Peach control No trace of oxalic acid.
The results of these experiments with oxalic acid show that
Sclerotinia cinerea when grown either on fruit juices or on
peaches produces more or less oxalic acid as a result of its meta-
bohsm. It is also significant that the other two fungi employed,
namely, Aspergillus and Penicillium^ which are not natural
parasites on the plum or the peach, produced no oxalic acid
tinder the conditions in which the experiments were carried out.
Summary
1. The brown-rot organism will infect fruits which are im-
mature, even penetrating those which are not more than half-
grown or those in which the pits are still soft, provided the
1914] ooo
COOLEY — SCLEROTINIA CINEREA oZ6
V
skin is punctured. Infection of green fruits is also effected when
a portion of the mycehal felt of the fungus is laid on the surface
of the plum. On the other hand, ripe or nearly mature fruits
may be readily inoculated by sowing a spore suspension on the
unpunctured surface.
2. The fungus does not show any particular aflTmity for the
middle lamella, but penetrates and permeates with equal avidity
any part of the host tissue.
3. A study of the effect of the organism on the host gives no
r^ncitUrp PAnrlpnnp thnt a toxic substance is abundantly secreted
ad
The fungus shows very slight cytolytic action with
m
o ^ow.i*..v.v. .x^... ; . the other hand, the
readily hydrolyzes cellulose from filter paper when
thFs is the only carbohydrate supphed. No general cytolytic
action of the organism on the cell wall of the host is perceptible.
5. An aqueous solution of pectin isolated from plums was co-
agulated by Sclerotinia, thus indicating the secretion of the
enzyme pectinase. In respect to its action on pectic substances,
Sclerotinia cinerea behaves in a manner similar to that of Pen-
icillium expansum, yet these two organisms produce very dif-
ferent effects on the host, the former producing a firm rot and
the latter a soft one. Neither organism will dissolve calcium
pectinate.
6. The experiments on the acid relations of the fungus mdi-
cate that the changing acidity of the host as the fruit reaches
maturity does not explain the fact that ripe fruit is more sus-
ceptible to the disease than green fruit.
7. The brown-rot fungus produces oxaUc acid when grown
either on a fruit juice medium or on peaches.
The writer takes pleasure in acknowledging his indebtedness
to Professor B. M. Duggar for his advice and helpful criticism
in this investigation. Part of this work was
summer of 1913 in the Laboratory of Plant 1
Universitv of Wisconsin
done during the
'athology of the
's to ftxnress his
Jones for the courtesy extended
him while at Madison.
Graduate Laboratory, Missouri Botanical Garden.
324 ANNALS or THE MISSOURI BOTANICAL GARDEN
[Vol. 1
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1914]
COOLEY — SCLEROTINIA CINEREA 325
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IVoL. 1, 1914J
326 ANNALS OF THE MISSOURI BOTANICAL GARDEN
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THE THELEPHORACE.E OF NORTH AMERICA. IP
Craterellus
EDWARD ANGUS BURT
Mycologist and Librarian to the Missouri Botanical Garden
Associate Professor in the Henry Shaw School of Botany of
Washington University
CRATERELLUS
Craterellus Pers. Myc. Eur. 2:4. 1825.— Fries, Epicr. 531.
1838; Hym. Eur. 630. 1874. — Saccardo, Syll. Fung. 6:514.
1888.— Ilennings, in Engl. & Prantl, Nat. Pflanzenfam. (i. 1**
127. 1898.
The type species of the genus is Craterellus cornucopioides
L. ex Pers.
Fructifications fleshy or membranaceous, pileate, often tubi-
form, infundibuliform, or flabelliform, sometimes clavate;
hymenium waxy-membranous, distinct, continuous, adnate to
the hymenophore, even or rugose; basidia simple; spores usually
white.
Craterellus is closely related by its fleshy C. Cantharellus,
C. odoratus, C. lutescens, etc., with the genus Cantharellus.
These species resemble so closely in coloration and habit species
of the latter genus that careful examination of the hymenium
should be made for generic determination. Craterellus has
its hymenium even or slightly rugose. In exceptional con-
necting species, such as C clavatus, it is somewhat lamelliform
for a part of the distance from margin of the pileus to the stem.
The clavate C. pisiillaris and C. unicolor connect Craterellus
closely with Clavaria.
Craterellus cornucopioides, C. ochrosporus, C. clavatus, C.
Cantharellus, and C. odoratus are edible species, which are often
abundant locally.
1 Issued September 30, 1914.
Note. — Explanation in regard to the citation of specimens studied is given in
Part I, Ann. Mo. Bot. Gard. i : 202, footnote. The technical color terms used in
this work are those of Ridgway, Color Standards and Nomenclature. Washington,
D. C, 1912.
Ann. Mo. Bot. Gakd., Vol. 1, 1914
5
(327)
[Vol. 1
328 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Key to the Species
Ilyinenium somewhat radiately lainelliform — at leaat near the margin j
stem solid ^
Hymcnium plane, rugose-wrinkled, or ribbed and rugose-wrinkled 2
1. Fructification large, 4-10 cm. high; stem about 1 cm. thick; spores 10-13 x
4_4i ^ .....i.e. clavatus
1. Fructification small, 1-li cm. high; stem 1 mm. thick; pilcus umbiHcate;
spores 9 X 7 M H- C. tklilescens
2. Fructification with pileus infundibuliform and pallid rose; hymenium
and stem white. In N. Carolina in moss near Kalmia bushes ... .4- C. roseus
2. Fructification entirely egg-yellowy about 3-9 cm. high, 2^-9 cm. broad ... 3
2. Fructification neither entirely egg-yellow nor with pileus pallid rose and
hymenium and stem white 4
3, Pileus convex, then depressed or nifundibuliform; stem solid. . .2. C. Caniharellus
3. Pileus convex, then depressed or cyathiform; stem hollow or cavernous;
fructification sometimes branched 3. C. odoiatus
4. Pileus tubiform with cavity extending nearly or quite to the base of
the stem 5
4. Pileus not tubiform, but instead infundibuliform. depressed, truncate,
convex, or flabelliform 6
5. Pileus and stem smoky brown to blackish; hymenium cinereous drab; spores
12-16 X 6-10 M 5. C. cornucopioides
5. Pileus drying avellaneous to snuff-brown; stem black with chamois-colored
pubescence at its base; hymenium chamois-colored or colored hke the pileus;
spores 12-15 x 7-8 m ^C. ochrosporus
5. Pileus somewhat tubiform; hymenium dark cinereous; spores 6-7] x 4J-5 m
7. C. duhius
6. Pileus somewhat tubiform or umbilicate, yellowish brown to fuscous; hyme-
nium and stem yellow; spores 10-12 x G-8 m 8.C. lulescens
6. Pileus infundibuliform, 2-3 cm. broad; hymenium pallid cinereous;
spores 10-12 x 6-7 m 9. C, sinuosus
6. Pileus deeply cup-shaped, 4-8 mm. broad; hymenium cream-buff;
spores 8 x 6 m ^0. C. calycuhis
6. Pileus convex, then umbilicate, 5 mm. broad; hymenium sometimes
obscurely lainelliform, chamois-colored; stem chamois-colored; spores
9 X 7 ix li' C'- delilescens
6. Pileus merely depressed, truncatCj convex, or clavate 7
6. Pilcus flabelfiform 8^
7. Fructification small, 1-3 cm. high, 4-9 mm. broad, narrowly obconic, white;
spores 3--1 fi in diameter 12. C. taxophilus
7. Fructification 2-5 cm. high, from obconic often becoming abrui)tly enlarged
and somewhat cerebriform at the upper end but with the stem remaining
comparatively slender IS. C. unicolor
7. Fructification large, 0-15 cm. high, clavate or obconic and truncate, tapering
downward; stem often bulbous at the base. Fructification dries sorghum-
brown to fuscous H' C. pistillaris
8. Pileus hgulate at first, then spreading laterally and becoming somewhat
palmately cleft into a few branches, fawn-color shading into bone-
brown. Knowm from Ohio 15, C. palmatus
1914]
BURT— THELEPHORACE^ OF NORTH AMERICA. II 329
8. Pileus somewhat triangular, drying a dirty pinkish buflf; hymenium
drying Isabella-color to day-color. Known only from Florida ..16.C. dilatus
8. Fructification entirely white; pileus reniform, dimidiate, attached
laterally to a slender erect stem. Known only from Washington
17, C. Humphreyi
I. Craterellus clavatus Pers. ex Fries, Epicr. 533. 1836-
1838. Plate 15. fig. 6.
Merulius clavatus Pers. Obs. Myc. i: 21. 18dQ .—C aniharellus
clavatus Fries, Syst. Myc. i: 322. 1821. —Nem^ophyllum clava-
ium Fries ex Patouillard, Tab. Anal. Fung, i : 193. /. 1^34. 1883-
18SQ. —Cantharellus hrevipes Peck, Rep. N. Y. State Mus.
33: 21. pi l.f, 18-20. 1879.
Illustrations: Schaeffer, Icon. Fung. pi. 164, 276.— Kromh-
holz, Abbild. und Beschr. pi. 45. f. IS-n.—FriGS, Sverig. Xtl.
Svamp. pi. Pi.— Richon et Roze, Atlas Champ, pi. 50. f. 10-1 4.—
Bresadola, Funghi Manger, pi. 5^.— Peck, Rep. N. Y. State
Mus. 33: pi. l.f. 18-20. —Harper, Mycologia 5: pi 93, 94.
Fructifications solitary or cespitose, fleshy, flesh whitish;
pileus narrowly obconic, turbinate, truncate or depressed, gla-
brous, ochraceous buff, attenuated into the stem, the margin
thin and erect; stem short, sohd, tomentose at the base; hyme-
nium lamelliform near the margin, rugose-wrinkled elsewhere,
becoming pruinose with the spores, light vinaceous drab, drying
drab; spores pale ochraceous in the mass, 10-13 x 4-4| /x.
Fructifications 4-10 cm. high; pileus 3-8 cm. broad; stem 1-2
cm. long, 8-15 mm. thick.
On the ground in coniferous woods. Maine to Connecticut
and west to Minnesota, and in Montana. July to September.
This species is intermediate between Craterellus and Cantharel-
lus. The marginal portion of the hymenium is like that of a
Cantharellus, and the remainder of the hymenium, like that of a
Craterellus. There is good authority for including this species
in Cantharellus and there is the authority of Fries and herbarium
usage for classing it in Craterellus. C. clavatus is edible but too
Specimens examined
be common in herb
Exsiccati: De Thuemen, Myc. Univ., 1807
Austria: G. Bresadola.
Maine: Sprague (in Curtis Herb., 5786).
[Voii. 1
330 ANNALS OF THE MISSOURI BOTANICAL GARDEN
New Hampshire: Shclburne, W. G. Farlow (in IMo. Bot.
Card. Herb., 4SG8).
Vermont: Lake Dunmore, E. A. Burt.
Connecticut : Rainbow, C. C. Hanmer, 1454 (in Hannicr Herb.).
New York: Ballston, C. H. Peck, the type of Cantharellus
hrevipes (in Coll. N. Y. State).
2. C. Cantharellus Schw. ex Fries, Epicr. 534. 183G-1838.
Plate 15. fig. 7.
Thelephora Cantharclla Schw. Schrift. d. Naturforsch. Gesell.,
Leipzig, i: 105. 1S22. —Cratcrcllus lateritius Berk. Grevillea
1 : 147. 1873.
Illustrations: Peck, Rep. N. Y. State Mus. 49: pi 44-
f. 1-5; Mem. N. Y. State Mus. 3^: pi 56. f. 17-21.— Hard,
Mushrooms /. 375.— Marshall, Mushroom Book 73. /.
Type: in Herb. Schweinitz.
Fructifications single or cespitose, fleshy, firm, egg-yellow;
pileus convex, becoming depressed or infundibuliform, glabrous,
yellow, the margin often lobcd or irregular; stem solid, cylindric
or tapering downward, glabrous, yellow; hymenium nearly
even or rugose wTinklcd, yellow, or with a reddish salmon tinge
and drying ochre-red; spores 7-10x3|-5| m-
Fructifications 4-9 cm. high; pileus 2^-8 cm. broad; stem
2|-5 cm. long, 5-10 mm. thick.
On the ground in open woods. Massachusetts to Alabama
and westward to Ohio; also in Mexico. June to September.
Abundant locally.
This species is so similar to Cantharellus cibarius in habit,
coloration, size and form— differing from the latter only in the
more even hymenium, that figures of C. cibarius will serve very
well for Craterellus Cantharellus, if allowance is made for the
different hymenium. The firm and solid stem of C. Cantharellus
distinguishes this species from C. odoratus easily. The latter
species sometimes has its pileus greatly branched. My illus-
tration of this species is photographed from the dried herbarium
specimen of the cotype of C. lateritius Berk. In this specimen
the lobes of the pileus were pressed together above before drying.
The hymenium of this specimen is now ochre-red and agrees
in color with that of the authentic specimen of C. Cantharellus
in Curtis Herb.; both these specimens have been poisoned. I
1914]
BURT — THELEPHORACE^ OP NORTH AMERICA. II 331
found the spores of the type in Herb. Schw. 8-9 x 3|-4 fx, or
a little slenderer than in northern specimens. Hard states
that the spores are yellowish or salmon colored in the mass
w
when collected. This species is edible.
Specimens examined:
Exsiccati: Ell. & Ev., N. Am. Fungi, 1921.
Massachusetts: Sprague (in Curtis Herb.); Milton, H. Webster.
Connecticut: East Hartford, C. C. Hanmer, 2391, 21^68 (both
in Hanmer Herb.).
Pennsylvania: West Chester, B. M. Everhart, Ell. & Ev., N.
Am. Fungi, 1921.
West Virginia : Eglon, C. G. Lloyd, 02292.
North CaroHna: Schweinitz, type (in Herb. Schweinitz);
Blowing Rock, G. F. Atkinson, 4313.
South Carolina: Clemson College, P. H. Rolfs, 1830.
Alabama: Peters (in Curtis Herb., 4539, and in Kew Herb.),
the ootype and type respectively of C. lateritius; Auburn,
F. S. Earle (in Mo. Bot. Card. Herb., 4928).
Ohio: A. P. Morgan (in Lloyd Herb.).
Kentucky: C. G. Lloyd (in Lloyd Herb.).
Mexico (?) : Botteri, 27 (in Curtis Herb.). If the stem is hollow
this specimen is C. odoratus.
3. C. odoratus Schw. ex Fries, Epicr. 532. 1836-1838.
Plates 15, 16. figs. 8-10.
Merulius odoratus Schw. Schrift. d. Naturforsch. Gesell.,
Leipzig, i: 91. 1S22. —Cantharellus odoratus Fries, Elenchus
Fung, i: 51. 1828.— C. confluens Berk. & Curtis, Jour. Linn.
Soc. Bot. 9: 423. 1867.
Type: in Herb. Schweinitz.
Fructifications gregarious, sometimes cespitose, simple or
branched, egg-yellow; pileus thin, convex, then depressed and
somewhat cyathiform, sometimes pervious, yellow, the margin
deflexed, often lobed or irregular; stem cylindric or somewhat
tapering towards the base, concolorous with the pileus, hollow
or cavernous; hymenium even or somewhat rugose-wrinkled,
ochraceous orange or with a reddish tinge approaching San-
ford's brown; spores even, 7-9 x 4-5 n.
Fructifications 3-7 cm. high; pileus 2-9 cm. broad; stem 2-4
cm
8 mm. thick
[Vol. 1
332 ANNALS OF THE MISSOURI BOTANICAL GARDEN
am
In moist places in woods. North Carolina and Georgia to
Ohio and Missouri. June to October.
Specimens of this species have sometimes been confused in
r
recent years with the better known C. Cantharellus, which ranges
farther north. The color and general habit of these species is
the same; both have the egg-yellow color and the characteristic
fragrance of Cantharellus cibarius when moistened after drying,
and all three are edible. Craterellus odoratus is more membra-
naceous than C. Cantharellus and it differs from both this species
and Cantharellus cibarius in having a hollow or cavernous stem
whose pliant walls may be pinched together, hke those of a
rubber tube, before the specimens are dried. Highly branched
forms may occur as shown in pi. 16 fig. 10a; this character was
unduly emphasized in the original description. The
collections in the Glatfelter Herbarium seem to show that Cra-
terellus odoratus is the most frequent Craterellus in the vicinity
of St. Louis. Dr. Glatfelter notes on his collection that he has
eaten this species and found it quite good. In pi. 15 fig. 8, I
give a figure, natural size, from a photograph of the dried her-
barium cotype of C. confluens B. & C., to show how close the
resemblance is to the specimens of C. odoratus, collected at St.
Louis and figured in the following plate. The type of C. con-
fluens has the hymenium rugose-wrinkled, as is often the case
in specimens of C. odoratus; its habit, dimensions, structure,
coloration, and spores are quite those of C. odoratus.
Specimens examined:
North Carohna: Salem, Schweinitz, type (in Herb. Schweinitz).
South Carolina: Society Hill, Ravenel, 192 (in Curtis Herb.).
Georgia: Station cited by Schweinitz.
Alabama: Auburn, L. M. Underwood.
r
Ohio: Oxford, L. 0. Overholts, 1721 (in Overholts Herb.).
Missouri: near St. Louis, N. M. Glatfelter, 348 (in Mo. Bot.
Gard. Herb., 42590), and /. B. S. Norton (in Mo. Bot. Gard.
Herb., 4926).
Mexico: near Orizaba, Botteri, 6 (type and cotype in Kew
Herb, and Curtis Herb., respectively, of C. confluens).
4. C. roseus Schw
1838.
Merulius roseus Schw. Schrift. d. Naturforsch. Gesell
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. II 333
zig, i: 91. 1S22. —Caniharellus roseus Fries, Elenchus Fung.
53. 1828.
Fructifications solitary, somewhat fleshy; pileus infundibuli-
form, somewhat strigose, pallid rose, the margin lobed and
inflexed; stem apparently stuffed, attenuated downward, white;
hymenium somewhat rugose, white.
In mosses, especially in proximity to Kalmia. North CaroHna.
Specimens of this species have the habit of Caniharellus
cibarius but are thinner. Fries received a specimen of Cratercl-
lus roseus from Schweinitz and expressed the opinion in 'Elen-
chus' that the species is good. I have seen no specimens of C.
roseus and base the above on the original description and the
comments by Schweinitz and Fries.
5
C. cornucopioides L. ex Pers. Myc. Eur
Plate 17. fi
Peziza cornucopioides L. Sp. PI. 1181. 1753. [1st ed.] —
Elvella cornucopioides Scop. FL Carn. 2: 476. 1760— Merulius
cornucopioides Pers. Syn. Fung. 491. ISOl. — Caniharellus
cornucopioides Fries, Syst. Myc. i: 321. 1821.
Illustrations: Vaillant, Botan. Paris, pi 13. f. S, 5.— Bolton,
Hist. Fung, pi 103.—¥lor. Dan. pi 384, :r^^0.— Holmskiold,
Fung. Dan. 2. pi J.— Sowerby, Brit. Fung, pi 74-— Schteffer,
Icon. Fung, pi ^^5.— Bulliard, Herb, de la France pi 150.—
Schnizlein, in Sturm, Deutsch. Flora 3: fasc. 31. pi 5.— Bresa-
dola. Fundii Manger. 75. pi 53.— Cooke, Brit. Edible Fung.
pi 11. f. 55.— Dufour, Atlas Champ, pi 70. /. ^57.— Hard
Mushrooms 451. /. 575.— Peck, Rep. N. Y. State Mus. 48: pi
24. f. 7-10.— d. Saccardo, Syll. Fung. 19: 478, for other refer
ences to illustrations.
somewhat cespitose; pileus thin
som
iwhat membranaceous, tubseform, pervious, sometimes
granular or minutely squamulose, smoky brown to blackish,
usually drying Front's brown, with the erect, spreading, or de-
curved margin generally lobed, wavy, or irregular; stem short,
hollow, even, blackish brown; hymenium even or rugose-
wrinkled, cinereous drab; spores hyaline, even, 12-16 x 6-10 n-
Fructification 5-8 cm, high; pileus 2^-5 cm. broad; stem 1-3
cm. Ions:, 3-5 mm. thick.
A'
061 ANNALS OF THE MISSOURI BOTANICAL GARDEN
On earth in mixed woods. Canada to South Carohna and
westward to Missouri. June to September.
The cornucopia craterellus is well characterized by its cornu-
copia-shaped or narrowly trumpet-shaped pileus ashy to sooty
brown in color, by thin flesh which is somewhat tough and
flexile, cinereous drab hymenium which sometimes has a brown-
ish tinge, and black stem. This species is too infrequent to
afford more than a few herbarium specimens in the regions where
I have collected fungi, but it is reported so plentiful in some
states as to be highly regarded as an edible species.
Specimens examined:
Exsiccati: Ravenel, Fung. Car. II. 27; Ellis, N. Am. Fungi,
321; Ell. & Ev., Fung. Col., 1723; Shear, N. Y. Fungi, 49^
Rabenhorst-Winter, Fung. Eur., 3640.
Sweden: L. Romell, 48.
Canada: /. Macoun, 72, 73,
Ontario: Casselman, J. Macoun, 347.
Vermont: Grand View Mt., E. A. Burt.
Massachusetts: Sprague, 211 (in Curtis Herb.).
Connecticut: W. A. Setchell.
New York: Sand Lake, C. H. Peck (in Coll. N. Y. State);
Alcove, C. L. Shear, Shear's N. Y. Fungi, 49; Ithaca, //
Schrenk (in Mo. Bot. Card. Herb., 4763, 42584), W. H.
Long, Jr., Ell. & Ev., Fung. Col., 1723.
New Jersey: Newfield, H. Leahy, Ellis, N. Am. Fungi, 321.
Pennsylvania: locality cited by Schweinitz, Syn. N. Am.
Fungi; W. Herbst (in Lloyd Herb.).
North Carolina: (in Curtis Herb., 502); locality cited by
Schweinitz, Syn. Fung. Car. Sup.
South Carohna: M. A. Curtis (in Curtis Herb.).
Ohio: Loveland, D. L. James, comm. by U. S. Dept. Agr.
Kentucky: Mammoth Cave, C. G. Lloyd.
Missouri: Perry ville, C. JL Dernetrio, Rabenhorst-Winter, Fung.
Eur., 3640; Meramec Highlands, P. Spaulding (in Mo.
Bot. Card. Herb., 4869).
6. C. ochrosporus Burt, n. sp. Plate 17. fig. 15.
An C. ocreatus Pers. Myc. Eur. 2 : 5. pi 13. f. 2. 1825?
Type: in Mo. Bot. Card. Herb., 42585.
Fructifications gregarious or cespitose; pileus thin, somewhat
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. II 335
membranaceous, tubaeform, pervious, minutely floccose-squam-
ulose, drying avellaneous to snuff-brown, the margin erect or
decurved; stem ghort, hollow, black, with chamois-colored
pubescence at the base; hymenium even or somewhat rugose,
sometimes colored like the pileus but in the type chamois-
colored: spores straw-3^ellow in the mass, even, obtuse. 12-15
X 7-8 ^.
Fructifications 4-7 cm. high; pileus 1-3^ cm. broad, 1-2 1
cm. long, 2-4 mm. thick.
On the ground among mosses in woods. New York and
Missouri. June to September. Probably abundant in Missouri.
Dr. Glatfelter noted a pleasant minty odor for the specimens.
This species closely resembles C. cornucopioides in form, but
differs from that species in having hymenium, spores, and base
of stem yellow. A collection from the same spot from which
the type collection came, but made in June two years later,
has the hymenium snuff-brown and approaches C. cornucopioides
in this respect. I am not aware of any data on C. ocreatus Pers.
except that based on the original description which is cited
above. That species has presumably not been collected by
European mycologists since the original collection from the
environs of Paris a century ago. Our specimens differ from
that description in having the stem yellow pubescent at the
base and the hymenium somewhat rugose, and they may differ
in other characters, e. g., spore colors, etc., not given in the
brief description of C. ocreatus. Hence I give to our American
specimens a distinct name.
Specimens examined:
New York: East Galway, E. A. Burt.
Missouri: Meramec Highlands, A^. M. Glatfelter (in Mo. Bot.
Card. Herb., 42585, type, and 42586-87); Columbia, B. M.
Duggar, 134-
7. C. dubius Peck, Rep. N. Y. State Mus. 31 : 38. 1879.
Illustrations : Hard, Mushrooms /. 380.
Type: in Coll. New York State.
Fructifications solitary or cespitose; pileus thin, infundi-
buliform or subtubiform, subfibrillose, dark brown or lurid
brown, pervious, the margin generally wavy and lobed; stem
short, hollow, colored hke the hymenium; hymenium dark
I Vol. 1
336 ANNALS OF THE MISSOURI BOTANICAL GARDEN
cinereous and rugose when moist, the obscure crowded irregular
wrinkles abundantly anastomosing, nearly even and paler when
dry; spores broadly elliptical or subglobose, G-7^ x 4^-5 ^t.
Fructification 5-7^ cm. high; pileus 2|-5 cm. broad, 4 mm.
thick.
On ground in woods. Ontario and New York to Illinois.
August to October. Rare.
The specimens of this species have the same coloration as
those of C. cornucopioides but differ from the latter in having a
shorter and more funnel-shaped pileus, and smaller spores.
Moffatt reported C. duhius as abundant at Glencoe, Illinois.
Specimens examined:
Ontario: Belleville, J. Macoun, 228 (in Coll. N. Y. State).
New York: Adirondack ^Its., C. //. Peek, type (in Coll. N.
Y. State).
Michigan: Sailor's Encampment, Univ. of Wis. Herb., 46.
8. C. lutescens Pers. ex Fries, Epicr. 532. 1838.
Plate 17. fig. 20.
Merulius lutescens Pers. Syn. Fung. 489. 1801; Albertini
& Schweinitz, Consp. Fung. 234. 1805. — Cantharellus lutescens
Fries, Syst. Myc. i: 320. 1821. — Merulius xanthopus Pers.
Myc. Eur. 2 : 19. pi 13. f. 1. 1825.
Illustrations: Vaillant, Botan. Paris. /)?. 11. f. 9, 10.
Scha3ffer, Icon. Fung. pi. 157. — Bolton, Hist. Fung. pi. 105. J. 2.
— Persoon, Myc. Eur. 2: pi. 13. f. :?.— Hennings, in Engl. &
Prantl, Nat. Pflanzenfam. (i.l**): 129./. 70 /f.— Stevenson,
Brit. Hym. 2:259.
Fructifications soUtary to cespitose; pileus thin, somewhat
membranaceous, varying from convex and umbihcate to tubi-
form or funnel-shaped, often pervious, yellowish brown to
fuscous, with margin often lobed or irregular; stem flcxuous,
cylindric, hollow, yellow, drying ochraccous buff, often hairy at
the base; hymenium remotely ribbed, even or rugose- wrinkled,
yellow, drying cadmium-yellow to ochraceous buff; spores
even, 10-12 x G-8 fx.
Fructifications 2-0-5 cm. high; pileus l|-3 cm. broad, stem
1^-4 cm. long, 2-4 mm. thick.
On moist ground in woods and swamps. Newfoundland to
North Carolina and westward to Michigan. August to October.
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. II 337
This species probably ranks next to C. cornucopioides in
frequency in the United States. The long and yellow stem
readily distinguishes this species from C. ochrosporus. Speci-
mens of Cantharellus injundihuliformis resemble those of Craterel-
lus lutescens in form, size, and color, but those of the former
species have true lamellse.
Specimens examined:
Exsiccati: Ellis, N. Am. Fungi, 1302; De Thuemen, Myc.
Univ., 404.
Sweden: Stockholm, L. Rornell, 49; Femsjo, L. Romell.
Newfoundland: Bay of Islands, A. C. Waghorne, 34 (in Mo. Bot.
Card. Herb.).
New Hampshire: Shelburne, W. G. Farlow, Ellis, N. Am. Fungi,
1302, and (in Mo. Bot. Card. Herb., 4932).
Vermont : Lake Dunmore, E. A . Burt.
Massachusetts: Worcester, G. E. Francis, 100.
New England: Sprague, 1689 (in Curtis Herb.).
New York: Sand Lake and Helderberg Mts., C. H. Peck (in
Coll. N. Y. State); East Galway, E. A. Burt.
Pennsylvania: locality cited by Schweinitz, Syn. N. Am. Fungi.
North Carolina: locality cited by Schweinitz, Syn. Fung. Car.
Sup.
Michigan: Glen Lake, C. G. Lloyd, 02462.
9. C. sinuosus Fries ex Fries, Epicr. 533. 1836-1838.
Cantharellus sinuosus Fries, Syst. Myc. i : 319. 1821.
Illustrations: Vaillant, Botan. Paris, pi. 11. f. 11-23.— Fries,
Icon. Hym. 2 : pi. 196. f. 2. — Dangeard, Le Botaniste 4 : 147. /. —
Gillet, Champ. France Hym. pi.
*
Fructifications ccspitose, slightly fleshy; pilcus infundibuli-
form, undulate and floccose, light drab; stem cylindric, stuffed,
pallid cinereous; hymenium at length with interwoven wrinkles,
pallid cinereous; spores 10-12 x 6-7 /i.
Fructifications 2-3 cm. high; pileus 2-3 cm. broad; stem l|-2
cm. long, 2-4 mm. thick.
On ground in mixed woods. South Carolina. Rare.
I have seen only dried herbarium specimens of Craterellus
sinuosus. The spore measurements are those of a specimen
from Sweden received from Romell. In this specimen the
hymenium has dried somewhat chamois-colored.
[Vol. 1
338 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Specimens examined :
Exsiccati: Rabenhorst, Fung. Eur., 208 (in Kew Herb.).
Sweden: L. Romell, 60.
South Carolina: Ravencl (in Curtis Herb., 2982).
C. crispus Fr., sometimes regarded as a variety of C sinu-
osus, was reported from New^ England, Sprague, by Berkeley &
Curtis, Grevillea i : 147, but the specimen is not satisfactory
for study. I do not, therefore, like to include C. crispus as
one of our species.
10. C. calyculus (B. & C.) Burt, n. comb.
Siereum calyculus Berk. & Curtis, Hooker's Jour. Bot. and
Kew Card. Misc. i: 238. 1849; Grevillea i: 161. 1873.
Type: type and cotype in Kew Herb, and Curtis Herb,
respectively.
Fructifications somewhat fleshy-membranaceous; pileus thin,
deeply cup-shaped, minutely tomentose, drying Saccardo's
umber, opaque ; stem apparently hollow, cream buff, attenuated
below, tomentose at the base; hymenium even or slightly venose,
cream buff; spores slightly yellowish under the microscope,
even, 8 x 6 a*.
Fructifications 2-3 cm. high; pileus 4-8 mm. broad; stem
ft
1 cm. long^ 1-2 mm. thick.
On ground in damp shady woods. North and South Caro-
lina. August and September.
Upon moistening, the type in Kew Herbarium proved too
soft and fleshy and the hymenium too waxy for a Stereum. The
sections have the structure of Craferellus, The speci(^s is near
C sinuosus and may prove to be a small form of this when
ample material gives more complete knowledge of the species,
but, for the present, I regard C calyculus as a distinct species.
I refer to C calyculus a collection made by Professor Atkinson
at Blowing Rock, North Carolina, the rough-dried and cespitose
specimens of which show a somewhat tubiform pileus and
spores 7-8 x 4| ti.
Specimens examined:
North CaroHna: Blowing Rock, G. F. Atkinson^ 1^200.
South Carolina : Santee River, Ravenel^ Curtis Herb., 1716
(the type and cotype in Kew Herb, and Curtis Herb.
respectively).
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. II
339
II.
C. delitescens Burt
17, fig. 18
Type: in Burt Herb.
Fructifications gregarious, cespitose, somewhat fleshy; pileus
thin, convex, then umbilicate, dry, fibrillose, sepia-colored, the
margin inroUed; stem equal, sohd, gLabrous, chamois-colored;
hymenium even or sometimes obscurely lamelliform, chamois-
colored; spores white, even, broadly ovoid, 9 x 7 m, borne four
to a basidium.
Fructification 10-15 mm. high; pileus 5 mm. broad; stem
10-15 mm. long, 1 mm. thick.
Growing among mos^s on vfery thin soil on rocks by water-
fall. Vermont. August.
This species is intermediate between Cantharellus and Craterel-
lus in its hymenial structure, but, as some of the specimens have
the hymenium even and bearing mature spores, I include the
species in Craterellus. The specimens are much smaller than
those of C. calyculus and have the pileus becoming
umbilicate. The little fructifications were well concealed
among the mosses; I have found them but once.
Specimens examined:
Vermont: Falls of Lana, Lake Dunmore, E. A. Burt, type.
12. C. taxophilus Thom, Bot. Gaz. 37: 215-19./. 1-8. 1904.
mere
Plate 17. fi
ns: Thom, ibid. f. 1-8.
Cornell Univ. Herb., 15445.
Fructifications single, rarely gregarious, fleshy - membra-
naceous, entirely white when young, becoming pallid to ochra-
ceous buff with age, drying cinnamon buff; pileus narrowly
obconic, slightly viscid, the apex truncate, plane, or depressed
and with a thin margin which is erect or expanded ; stem solid,
equal or tapering downward, flexuous, pruinose, with scattered
white hairs at the base; hymenium even, becoming longitudi-
nally rugose-WTinkled with age or upon drying; spores white,
even, subglobose, 3-4 m in diameter, borne four to a basidium.
Fructifications 1-3 cm. high; pileus 4-9 mm. broad; stem \-2
cm. long, ^-1 mm. thick.
On rotten twigs and leaves under prostrate branches of Taxus
canadensis. New York. October and November.
This delicate fungus was under observation by Dr. Thom
[Vol. 1
340 ANNALS OF THE MISSOURI BOTANICAL GARDEN
for a month and is described in detail and beautifully illustrated
in connection with his original description in tlie work cited
above. I reproduce merely some simple outline sketches of
C. taxophilus; this is a very distinct species. The specimens
were found in Fall Creek Gorge and nowhere except under
prostrate branches of Taxus, yet they grew on rotting twigs
and leaves of other species as well as on pieces of Taxus.
Specimens examined :
New York: Ithaca, C. Thorn, Cornell Univ. Herb., 15445.
13. C. unicolor Rav. Grevillea i: 148. 1873.
Plate 16. fig. 11, 12.
C. corrugis Peck, Bull. Torr. Bot. Club 26: 09. 1899.
Type: in Ravenel, Fung. Car. II. 2G.
Fructifications solitary or ccspitose, fleshy, with the flesh
white, soft, soon shrinking and leaving the pileus hollow;
pileus at first clavate, obtuse, flesh-colored tinted with violet,
soon obconic or turbinate, broadly convex or truncate, and
often abruptly cercbriform at the upper end, glabrous, ochra-
ceous buff, drying Rood's brown to Natal-brown, the margin
obtuse, corrugated by the hymenial \ninklcs; stem short, equal
or tapering downwards, colored like or a little paler thiui the
pileus; hynienium wrinkled or corrugated, colored like the
pileus; spores white, 8-12 x 4-G m.
Fructifications 2-5 cm. high; pileus l§-5 cm. broad; stem 1-2 1
cm. long, 5-8 mm. thick.
On ground in thin woods. Massachusetts, Pennsylvania,
and South CaroHna. October to January.
This fungus presents strikingly the vagaries in the distri-
bution of fungi. It was originally collected at Black Oak,
South Carolina, in 1850, by Ravenel, in sufficient nuantitvso
distributed
Appar
ently, this fungus, whenever collected, was referred to other
species until 1898, when members of the Boston IVIycological
Club found it in several localities in Massachusetts and it was
adequately described by Peck, as C. corrugis, from specimens
received from Dr. Francis. I have received no specimens of
this species since that season; I searched for it in vain for several
years in the adjoining state, Vermont. I have compared the
specimens of C. corrugis, received from Dr. Francis, with Peck's
1914]
BURT— THELEPHORACE^ OF NORTH AMERICA. II 341
type and with the specimens of C. unicolor in five different
copies of Ravenel's 'Fungi Caroliniani.' C. corrugis is certainly
the same species as C. unicolor. It is very strange that in the
interval of nearly half a century from the time of the original
collection, C. unicolor did not attract attention from an inter-
mediate station.
Specimens examined:
Exsiccati: Ravenel, Fung. Car. II. 26; Ell. & Ev., N. Am.
Fungi, 1922a under the name C. pistillaris.
Massachusetts: Worcester, G. E. Francis, 61, 84, and col-
lection dated Nov. 2, also the type (in Coll. N. Y. State)
of C. corrugis; Lynn, H. Webster; Medford, Mrs. Page and
Mrs. De Long, ex Herb. Boston Mycological Club, 420;
Arlington Heights, E. A. Burt.
Pennsylvania: Trexlertown, W. Hcrhst, the C. clavatus of his
Tungal Flora'; West Chester, B. M. Everhart, Ell. & Ev.,
N. Am. Fungi, 1922a.
South Carolina: Black Oak, Ravenel, 14-06 (in Curtis Herb, and
in Kew Herb.), and type, Ravenel, Fung, Car. II. 26,
14. C. pistillaris Fries, Epicr. 534. 1836-1838.
Plates 16, 17. figs. 13, 14.
Illustrations: Schfcffer, Icon. Fung. pi. i^P.— Harper, Myco-
logia 5: 263. pi. 95.
Fructifications gregarious, fleshy-spongy, drying sorghum-
brown to fuscous; pileus somewhat clavate to turbinate or
narrowly obconic, truncate, or somewhat convex, at first yel-
lowish cinnamon, then becoming tinged with fuscous, the edge
obtuse; stem solid, paler than the pileus, often bulbous at the
base; hymenium corrugated and rugose-wrinkled, colored like
the pileus, drying sorghum-brown to fuscous; spores even, 10-
X 6-8 IX.
Fructifications 6-12 cm. high; pileus 2-3 1 cm. broad; stem
3-6 cm. long, 4-12 mm. thick.
On ground in woods under coniferous trees. New Hampshire,
Vermont, and Michigan. August to October.
Specimens of this species have so nearly the coloration of C.
unicolor that those, small and undeveloped, in a collection of
C. pistillaris cannot readily be distinguished from partially
developed specimens of C. unicolor; but with age, those of C.
12
[Vol. 1
342 ANNALS OF THE MISSOURI BOTANICAL GARDEN
unicolor—oT at least some of them— have the pileus enlarge
abruptly in diameter near the upper end and become abruptly
globose-ccrebriform on a slender stem, as shown in figs. 11 and
12, while C. pistillaris increases in length as well as in diameter,
tapers downward more uniformly from the truncate upper endi
and may have the stem bulbous at the base.
It is a vexed question with mycologists whether Craterellus
pistillaris Fr. is Clavaria pistillaris L. The specimens which I
refer to Craterellus pistillaris agree well with specimens of this
species in Curtis Herbariuju, collected at Upsala, Sweden, in
1853, and communicated by E, P. Fries. PL 16 fig. 13 is from a
photograph, natural size, of these specimens. Their spores are
9 X G /x. The Friesian specimens have the same dark color as
our American specimens. Only one of the former shows a
bulbous tendency at the base of the stem; in this respect our
specimens are more like the illustration of Scha^ffer, cited above.
I believe, therefore, that we have Craterellus pistillaris Fr. in
our flora. I have collected in mixed frondose woods in Mis-
souri what I refer to Clavaria pistillaris as understood by Euro-
pean mycologists. As compared with the former species it is
of softer structure, much paler in color, more regularly clavate
sometimes splitting at the apex. The illustrations of
ropean authors agree well in regard to Clavaria pis-
tillaris. The colored figures of this species in Batsch, Bulliard,
Sturm, Dufour, Flora Danica, Hussey, Krombholz, Quelet,'
and Sowerby present fructifications of the same habit and
bright coloration which we have by Peck, Bull. N. Y. State
Mus. 94: pi 93. f. 1-4. and I\Icm. N. Y. State Mus. 4: pi. 66
/. 15-17.
Specimens examined:
Sweden: Upsala, E. P. Fries (in Curtis Herb.).
Austria: G. Brcsadola.
New Hampshire: Shclburne, W. G. Farlow (in Mo. Bot. Card
Herb., 4933).
Vermont: Middlebury, E. A. Burt.
15. C. palmatus Burt & Overholts, n. sp. Plate 17. fig. 19,
Type: in Mo. Bot. Card. Herb, and in Overholts Herb
most
Fructificat
gregarious or perhaps cespitose, fleshy-soft
shading into bone-brown towards the stem
1914]
BURT — THELEPHORACEiE OP NORTH AMERICA. II. 343
glabrous, flattened and ligulate at first, then spreading out
laterally at the apex, and at length somewhat palmately cleft
into 2-12 unequal, obtuse, finger-shaped branches; stem curved,
solid, equal or somewhat tapering towards the base, bone-
brown, sometimes swollen where attached to the substratum;
hymenium even or but sUghtly venose, inferior, colored like
the pileus; spores white, even, pyriform, tapering to the base,
6-8 X 3-4 M.
Fructifications 1-2| cm. high; pileus 3-15 mm. broad, 1
mm. thick; stem 8-15 mm. long, 1-1 1 mm. thick.
On rotten chunks of wood in frondose woods. Ohio. June.
All specimens of the collection except one have the pileus
flabelliform; in this exceptional specimen, the pileus is narrowly
turbinate, depressed, and with the finger-shaped branches
arranged in a circle on the margin, pi. 17 fig. 19b. This species
makes for Craterellus the same connection between the central-
stemmed, cup-shaped type of pileus and the flabeUif orm type
that Thelephora multipartita shows in Thelephora, and that is
common in Stereum. The hymenium of the flabelhform speci-
mens of Craterellus palmatus is so similar to the upper surface
of the pileus in color and consistency that one cannot readily
distinguish between these surfaces in the dried specimens.
For these reasons, the present species cannot be referred to
either Skepperia or Friesula, and it is of especial interest in
showing that Craterellus has a natural section of species with
flabelliform pileus. The spores of C. palmatus are noteworthy.
Specimens examined:
Ohio: Oxford, L. 0. Overholts, 1649, type (in Mo. Bot. Gard.
Herb, and in Overholts Herb.).
i6. C, dilatus Burt, n. sp. Plate
Type: in Farlow Herb.
Fructifications single, fleshy; pileus flabelliform, somewhat
triangular, glabrous, drying a dirty pinkish buff, the margin
somewhat irregularly lobed, crisped, and curving upward; stem
solid, equal, flexuous, drying Natal-brown, with white myce-
Hum at the base; hymenium even, drying Isabella-color to clay-
color; spores white, even, broadly ovoid, obtuse, 8-10 x 6-7 /x.
Dried fructification 4 cm. long; pileus 15 mm. long, 15 mm.
broad, | mm. thick; stem 2^ cm. long, hardly 1 mm. thick.
6
17. fig. 16
[Vol. 1
344 ANNALS OF THE MISSOURI BOTANICAL GARDEN
On sandy ground in swamp. Florida. September.
Only a single fructification was collected; the description is
based upon this dried specimen. The species is distinguished
by its fan-shaped, triangular pileus and the comparatively long
and slender stem. Its characters are those of a true Craterellus
and yet such that we cannot regard it as a flabellate form of any
other species-
Specimens examined:
Florida: Sorrento Swamp, R. Thaxtcr, type (in Farlow Herb.).
17. C. Humphreyi Burt, n. sp. Plate 17. fig. 22.
Type: in Burt Herb, and in Humphrey Herb.
Fructifications gregarious, fleshy, moderately tough and
flexible, entirely white, usually with the pileus standing out
horizontally at the apex of the erect stem; pileus reniform,
dimidiate, sometimes clasping behind, convex, becoming plane
or somewhat depressed, usually even, dry, minutely pubescent,
the margin entire, even or slightly crisped; stem lateral, erect,
often bent at right angles just before joining the pileus, cylindric
below, equal, solid, pubescent; hymenium nearly even, some-
times radiately venose near the stem, brittle when fresh;
spores white, even, subglobose, 3|-4^ x 3| tx.
Fructifications 3-7 cm. high; pileus 6 mm. - 2 cm. long, 1-3 1
cm. broad, f mm. thick; stem 2|-6 cm. long, 2 mm. thick.
On humus and among mosses in low swampy thicket. Wash-
ington. October.
The habit of this curious species is very suggestive of Hydnum
auriscalpium; many of the specimens have the erect stem bent
at right angles near the apex so that the pileus extends out in a
horizontal plane. Sometimes the stem branches at its upper
end and bears two pilei. The pubescence on the stem is rather
coarse and is most abundant towards the base. All parts of
the fructification were rather brittle in vegetative condition,
and broke when bent too far. It is a connecting species be-
tween Craterellus and Arrhenia, but with the hymenium rather
too even for Arrhenia, in my opinion.
Specimens examined:
Washington: Hoquiam, C. J. Humphrey, 1386, type.
1914]
BURT — THELEPHORACEiE OF NORTH AMERICA. II 345
Berkeley & Curtis, Jour. Linn. Soc. Bot. lo: 328, described
three species of Craferellus from Cuba, which have been trans-
ferred to other genera by Patouillard, Bull. Soc. Myc. France
15: 193-94. pi 9, as follows: C spathularius to Skepperia
and C. marasmioides and C. pulverulentus to Cymatella. I have
received no collections referable to these genera and defer
their consideration to the final part of my monograph in the
hope that some specimens may be received in the meantime.
Craterellus canadensis Kl. ex Saccardo, Syll. Fung. 6: 519.
1888, was published by Berkeley, Ann. Nat. Hist. 3:380. 1839,
under the name Cantharellus canadensis Kl. from a specimen in
Hooker Herb, bearing manuscript notes by Klotzsch. The
specimen was collected in Canada by Richardson. In connec-
tion with the original description, Berkeley noted that the
nearest affinities of C. canadensis are with C. clavatus. In 1856,
after studying the specimens in Herb. Schweinitz, Berkeley &
Curtis, Jour. Acad. Nat. Sci., Phila. N. S. 3: 206. 1856, note
that Cantharellus canadensis Kl. is apparently the same species
as Cantharellus floccosus Schw. I have seen no specimens of
C. canadensis and follow Berkeley's final disposition of the
species.
(To be continued.)
\
[Vol. 1, 1914]
346 ANNALS OP THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 15
All figures of this plate have been reproduced natural size from photo-
graphs of dried herbarium specimens.
Fig. 1. Thelephora cmspitulam. From authentic specimen in Curtis Herb., col-
lected by Schweinitz in North Carolina.
Fig, 2. TAutosa, From authentic specimen in Curtis Herb., collected by Schwei-
nitz in North Carolina.
Fig. 3. T, dentosa. From cotype in Curtis Herb., collected in Cuba by C.
Wright.
Fig. 4. T. perplexa. From type in Curtis Herb., collected in Cuba by C Wright,
238. a shows a rcsupinate portion, and 6, an ascending portion of the specimen.
Fig. 5. T. comucopioides . From specimen collected in Castleton Gardens,
Jamaica, by F. S. Earle, 238.
Fig. 6. Craterellus clavatus. From specimen collected at Lake Dunmore, Vt.
Fig. 7. C. Cantharellus, From the cotype in Curtis Herb,, 4539, of C, lateritius^
collected in Alabama, by Peters.
Fig. 8. (7. odoralus. From the cotype in Curtis Herb, of C. confluens, collected
near Orizaba, Mexico, by Botteri, 6.
Fig. 9. C. odoratus. From the specimens in Curtis Herb., collected at Society
Hill, S. Carohna, by Ravenel, 192.
A.N^. Mo. JjOT. Gaud.. Vol. 1, 1011
Plate 15
2
a
8
1
w***
I
¥^
\\
liURT— TIIETJ:riIURACEAE OF NORTH AMERICA
1. TllKLKrHOKA OAESriTULANS. — 2. T. LUTOSA. — ;^. T. OENTOSA.— 4. T. FKUTLKXA
5. T. COr.NUCOrTOIDES. — 0. CUATEllELLUS CLAVATIJS. — 7. C. CANTIIARELLUS.
ANi> -.». C. OnOKATUS.
COCKAYNE, L'.USTO.\
348 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1, 19141
Explanation of Plate
PLATE 16
All figures of this plate have been reproduced natural size from photo-
graphs of dried herbarium specimens, but in the case of fig. 10 the
specimens were moistened.
Fig. 10. C. odoratus. From specimens collected near St. Louis, Mo., by N. M.
Glatfelter, 348. The rough dried specimens were moistened before being photo-
graphed, a shows a branched specimen; I, a fructification spHt longitudinally to
show extent of depression of the pileus and the hollow stem; c, view of hymenium.
Fig. 11. C. unicolor. From authentic specimen in Curtis Herb., collected at
Black Oak, S. Carolina, by Ravenel, 1406.
Fig. 12. C. unicolor. From specimen of C. corrugis collected at Medford, Mass.,
by Mrs. Page and Mrs. DeLong.
Fig. 13. C. pistillaris. From specimen in Curtis Herb., collected at Upsala,
Sweden, by E. P. Fries.
Ajsn. Mo. Box. Gakd., Vol. 1, 1014
rLATK 16
y
BURT — THEi.KlMIORACKAK OF NORTH AMERICA
10. CKATEUKLLUS ODORATUS. — 11 Anu 12. C. UMCOLOK. — 13. C. TlKriLLAUlS
COCKAVNK, HOSTON
[Vol. 1, 1914]
350 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 17
All figures are natural size. Figure
um
photographed in case of specimens used for figs. 15 and 17.
Fig. 14. C, pistillaris. From specimen collected under hemlock (Tsuga) tree, at
Middlebury, Vt.
Fig. 15. C. ocJirosporus. From type specimens in Mo. Bot. Gard. Herb., col-
lected near St. Louis, Mo., by N. M. Glatfclter, 1253. a is spht longitudinally to
show the depth of depression of the pileus; 6, side view.
Fig. 16. C. dilatus. From type in Farlow Herb., collected at Sorrento Swamp,
Florida, by R. Thaxter. a shows upper surface of pileus, and ft, the hymenium.
Fig. 17. C. comucopioides. From specimen collected in Canada, by J. Macoun
72.
Fig. 18. C. delitescens. From type specimens collected at Lake Dunmorc, Vt.
Fig. 19. C. pabnalus. From type specimens in Mo. Bot. Gard. Herb, and Over-
holts Herb., collected at Oxford, Ohio, by L. O. Overholts, 1649. a shows specimens
having flabelliform pileus, and 6, a specimen with turbinate pileus.
Fig. 20. C. lutescens. a shows hymenium of specimen collected at Shelburne,
w
Lake
Fig. 21. C. taxophilus. From sketches of photographs of type specimens when
in vegetative condition, collected at Ithaca, New York, by C. Thorn.
Fig. 22. C. Humphreyi. From sketches of the type specimens when in vegetative
condition, collected at Hoquiam, Wash., by C. J. Humphrey, 1386.
Ann, Mo. Bot. Gaud., A^ol. 1, 1914
Plate 17
^3?^^
a
1>
20
21
BURT — TIlia.r:ril()RA('KAE OF NORTH AMERICA
14. CRATKKKLT.rS IMSTILLAIMS. — 15. C. OCIIUOSrOUUS.
C\ DELITESC^ENS.- li>. C. TALMATUS
17. C. COUNUCOriOIDKS.- 18
21. C. TAXOrHlLUS. — 22. C
IG. C. DILATT'S.-
-20. C. LUTEfet KNS
HUMPIIUEYI.
rUCKAYNP:, HUSTON
THE EFFECTS OF SURFACE FILMS ON THE RATE OF
TRANSPIRATION: EXPERIMENTS WITH POTTED
POTATOES
B. M. DUGGAR
Physiologist to the Missouri Botanical Garden, in Charge of Graduate Laboratory
Professor of Plant Physiology in the Henry Shaw School of Botany of
Washington University
AND J. S. COOLEY
Formerly Rufus J. Lackland Fellow in the Henry Shaw School of Botany of
Washington University
In a previous report^ we have presented data which is be-
lieved to justify the conclusion that an application of a surface
film of Bordeaux mixture to the leaves of the castor bean or
the tomato increases materially the rate of transpiration. The
importance of a careful determination of various physiological
effects of this spray mixture was suggested primarily by the
increased vitality and yield exhibited by potatoes {Solanum
tuberosum) treated with this fungicide during seasons when
fungi and insects were unimportant factors. In our previous
experiments the potato was not included, and it seemed most
important, as a next step, to ascertain the effects of certain
sprays upon the transpiration of this plant.
Experience has demonstrated that the potato may not be
used satisfactorily in potometer experiments. Moreover, it
was desired to arrange the experiment so that the transpiration
quantities obtained might represent an interval of a week or
more. On the other hand, it had been found as a result of our
previous work with potted tomatoes that a very considerable
amount of labor is required when it becomes necessary to add
measured quantities of water every day to a series of fifty or
more potted plants. Accordingly, for this and for other work
proposed, a method was devised whereby we were able to employ
a self-watering device based on a principle often used in the
laboratory.
^ Duggar, B. M., and Cooley, J. S. The effect of surface films and dusla on the
rate of transpiration. Ann. Mo. Bot. Gard. 1:1-22. pL L 1914.
Ann. Mo. Bot. Gard., Vol, 1, 1914
(351)
[Vol. 1
352 ANNALS OF THE MISSOURI BOTANICAL GARDEN
The apparatus is shown in pi. 18. The rack, or support,
is made of a single sheet of galvanized iron 18 cm. wide and 55
cm. long, these dimensions being adequate for a stand 33 cm.
high. Besides cutting a hole in the upper part for the insertion
of the neck of the bottle, the operation of making a stand will
be clear from the plate and involves merely a few slits with the
shears, the balance being accomplished by bending. Two or
four rivets may be used if additional strength is required. With
regard to other features of the apparatus it is well to note that
(1) the shoulder of the flower pot rests on the rim of a tin cup
somewhat deeper than the pot, the latter containing the im-
mediate supply of water; (2) there is an inverted bottle with a
capacity of about 1500 cc. serving as a reservoir of water and
aspirator ; and (3) the bottle is connected with the cup by glass
and rubber tubing.
In setting up an experiment the exposed area of the pot
(above the shoulder) and the soil are covered with paraffin or
parawax; the cup is filled with water to such height that when
the pot is inserted the water will rise to the height of about
2 cm. on the side of the pot, thus insuring adequate absorption;
while a notch in the side of the cup makes it possible to introduce
the rubber tube connecting with the bottle, this tube being
adjusted to reach just below the new level of water in the cup.
With a tube of proper diameter, the water level in the cup is
kept practically constant so long as the bottle contains water.
This apparatus, complete, may be quickly and sufficiently
accurately weighed on the Troenmer scales. To prevent up-
setting, after arranging in the experimental area, it is well to
make the stand secure by providing a small hole in the base,
through which a bamboo stick may be thrust into the soil. To
this stake, also, for further support, the bottle may be fastened
by cord or rubber band.
The device above described has saved much time and has
enabled us to obtain a soil moisture content practically uniform
in all the pots used in the experiment. It possesses the disad-
vantage of tending to maintain a moisture content which ^or
long-term cultures is too high for the best growth of the potato.
A slight modification of the method would seem to be practi-
cable in several aspects of transpiration work.
1914]
DUGGAR AND COOLEY — FILMS AND TRANSPIRATION 353
Three weeks before the experiment began the plants were
repotted, new 5-inch pots of good quality being used, and at
the time of the installation of the experiment the drainage holes
in the pots were carefully corked, so that all transfer of water
would be through the porous walls. The potato plants em-
ployed were grown in the greenhouse during the early spring,
but on April 20, about two weeks before the test was made, they
were placed outside, to insure hardiness. When used, the
plants were from 25 to 45 cm, high, each plant with from about
15 to 30 leaves. Some plants were blossoming, and tubers
were forming.
The experiment embraced 7 series, or lots, of 10 plants each,
sprayed with mixtures as follows: (1) strong Bordeaux, (2)
control, no spray, (3) weak Bordeaux, (4) lime wash, (5) lime
sulfur, (6) strong Bordeaux and lampblack, and (7) lime wash
and lampblack. The strong Bordeaux (designated hereafter
Bordeaux) contained 12 grams CuS04 and 14.4 grams CaO
per liter of water, being approximately the 5-6-50 formula of
agricultural practise. It was made up in the usual way. The
weak Bordeaux was one-half the strength of the stronger
mixture. The lime wash was a Ca(0H)2 suspension consisting
of 60 grams of CaO per liter of water. A commercial prepara-
tion of lime sulfur was used, and this was diluted, as usual, to
about 1-25. The Bordeaux-lampblack and the lime-wash-
lampblack preparations were made by rubbing into small
quantities of the Bordeaux and lime wash 5 and 10 grams re-
+
spectively of lampblack, then diluting to one liter.
The method of selecting the plants for the different lots was
precisely that described in the previous report; that is, selecting
at one time 7 plants (as many as there were lots) between which
there could be little or no choice, and distributing these at
random, 1 to each lot until each included 10 plants. All plants
(except controls) were sprayed on May 5, but a rain that night,
before protection was provided, necessitated respraying the
following day. After spraying, the plants were placed on the
stands and each connected
They were
arranged on an exposed lawn, each lot occupying a row, with
the plants 4 feet apart. Moreover, several rows of potted pota-
toes were arranged around the entire area in order that all
[Vol. 1
354
ANN
OF THE MISSOURI BOTANICAL GARDEN
plants in th'- oxperimental area might have equal exposure
Over the
whole ari
ntal plot a frame was provided, so that the
be protected by tarpaulins in case of rain.
Fortunately, however, no rain occurred during the period of the
experiment.
After a prehminary exposure of 24 hours, which enabled us to
determine that the 70 plants of the experimental area were in
good condition, the initial weighings were made. A definite
order was estabhshed, this being crosswise of the different lots.
The same order was observed at the close of the period, and
similarly in the second period a consistent scheme was followed,
in order that the time interval might be as uniform as possible.
After the weighings at the close of the first period, all plants
were discarded which showed any signs of weakness or injury
of the experiment. It should be
ons were taxing. The weather
from the condit
stated, too.
these condit
was bright and warm, the pots were severely exposed, and, as
already noted, the water content of the pots was necessarily
fairly high. With the plants remaining in a condition appar-
ently normal and vigorous from the first period, a second
"run" was made, the latter including from 4 to 7 plants in
TABLE SHOWING WATER LOSS AND GREEN WEIGHT OF THE PLANTS
Lot
1st period, May 6-11,
10 plants
1
2
3
4
5
6
7
Film covering
Ave.
1
water
loss per
plant
Bordeaux, strong
520. 6
Control
413.8
Bordeaux, weak
642.4
Lime wash
584.5
Limo sulfur
443.0
Bordeaux and lamp-
black
792.1
Lime and lampblack
596,6
Ave.
green
weight
per plant
Water
loss per
g., green
weight
2nd period, May 11-15,
7 plants
Ave.
water
loss per
plant
50.3
61.0
60.9
70.7
62.8
66.1
58.3
10.46
6.78
10.54
8.27
7.06
463.3
433.3
574.0
613.6
450.7
11.97
10.20
653.0
585.6
Ave.
green
weight
per plant
Water
loss per
g ., green
weight
55.0
63.1
61.7
76.3
70.0
75.2
66.8
8.42
6.8G
9.30
S.Ol
0.44
8.69
8.78
each lot. In selecting plants for this second period, the size
factor was again taken into consideration, as far as possible.
1914]
DUGGAR AND COOLEY— FILMS AND TRANSPIRATION
355
More stress should, however, be laid upon the data from the
first period. The green weights of the plants discarded at the
close of the first period were taken immediately, while those
plants used in the second period could not be weighed until the
close of that interval. This small interval of time, however,
could cause no material change in the weights. In the accom-
panying table there are given in grams the average water loss
per plant, the average green weight per plant, and the water
loss per gram of green matter.
From the data exhibited it is obvious that with potted
potatoes, as with castor bean leaves and potted tomatoes in our
earlier experiments, there is a marked acceleration of transpira-
tion induced by spraying with Bordeaux mixture, as also with
some other films. Of the several films employed, lime sulfur
alone yields an average water loss comparable with that of
unsprayed plants. Of all lots showing increased transpiration
those treated with weak Bordeaux and lime wash were in some
respects most satisfactory, inasmuch as the plants used, like
those in the control, were, in general, in very good condition
throughout the period of the experiment. On the other hand,
those treated with the stronger Bordeaux, the Bordeaux and
lampblack, and the Hme and lampblack gave, towards the close
of the periods, evidences of the injurious effects of the increased
transpiration (apparently) upon the vitality of the plants.
These statements may not seem to be in entire accord with
the figures presented, for during the second period of the ex-
periment, for example, the transpiration quantity is relatively
greatest in the case of those plants sprayed with weak Bordeaux
mixture. Nevertheless, our observations enabled us to predict
that certain lots, especially numbers 1 and 6, would give in the
second period, particularly, transpiration values less than
might be anticipated. The smaller quantities in the lots re-
ferred to, as contrasted with the weak Bordeaux, are to be
explained, in fact, as a direct result of incipient wilting and
sHght injury, brought about by the higher transpiration capacity
induced under conditions already accentuating transpiration.
It is believed, in the first place, that the experiments here
reported confirm our earlier conclusion, namely, that a film of
Bordeaux mixture facilitates water loss; but, in the second place,
o-p rV'OL. 1, 1914]
OiiO ANNALS OF THE MISSOURI BOTANICAL GARDEN
treatment with a fairly thick lime wash or lime wash and lamp-
black also increases the transpiration rates, the latter more than
the former. Lampblack added to Bordeaux seems also to give
a higher rate than the Bordeaux alone. It is to be emphasized,
however, that the strength of the lime wash employed is four
times as great as the lime in the stronger Bordeaux mixture;
likewise, more lampblack is used with the lime wash than with
the Bordeaux. It seems to be definitely established that cer-
tain specific characters of the film are important, but these
results suggest, further, that the additional quality of color is
a factor requiring consideration. The fact that injury may
result from the accelerated transpiration induced by a heavy
film of Bordeaux under the conditions of our experiment does
not mean that under normal conditions of growth in the field a
benefit may not accrue to certain plants-
ciated with a high transpiration rate.
Graduate Laboratory, Missouri Botanical Garden.
from factors
EX5>LA NATION OF PlATE
PLATE 18
View of the apparatus (with tomato plant) by means of which watering
was automatically controlled. It has been found convenient to have
both stand and cup painted green. For description see text, p, 322.
A->'N. Mo. Hot. Gard., Vol. 1, Vji4
Plate 18
J-
DUCGAR AND CUOi.KY — TRANSTIRATION
COOKAV^i: BOSTON
Annals
of the
Missouri Botanical Garden
*
Vol. I NOVEMBER, 1914 No. 4
THE THELEPHORACE.E OF NORTH AMERICA lll^
Craterellus borealis and Cyphella
EDWARD ANGUS BURT
Mycologist and Librarian to the Missouri BotaJiical Garden
Associate Professor in the Henry Shaw School of Botany of
Washington University
Since the publication of Part II, on Craterellus, Dr. Farlow
has very kindly called my attention to, and permitted me to
study, a specimen of a rare species from Labrador which was
not included in my account of our North American species.
This species is now described here so as to bring its description
and illustration continuous with those of our other species of
Craterellus.
The following is suggested for insertion in *'Key to the
Species," on page 328 (Ann. Mo. Bot. Gard. i: 328. 1914).
6. Pileus membranaceous, iiifundibuliform, pale buff; hymenium pale buflf; spores
5-7x4-51^1; from Labrador. See page 357 (Ann. Mo. Bot. Card. i:357,
1914) C. borealis.
Craterellus borealis Burt, n. sp. Plate 19. fig. 1.
Type: in Farlow Herb.
Fructifications solitary, small; pileus infundibuliform, taper-
ing uniformly to the stem, glabrous, drying between cartridge
buff and cream-buff, the margin entire; stem nearly equal,
Note. — Explanation in regard to the citation of epecimeiis studied is given in Part
I, Ann. Mo. Bot. Gard. i:202. 1914, footnote. The technical color terms used in
this work are those of Ridgway, Color Standards and Nomenclature, Washington,
D. C, 1912.
* Issued January 30, 1915,
Ank. Mo. Bot. Qabd., Vol. 1, 1914
(357)
[Vol. 1
358 ANNALS OF THE MISSOURI BOTANICAL GARDEN
slender, minutely downy, pale mouse-gray; hymenium colored
like the pileus, remotely ribbed, with the ribs radiating from
the stem, thin, branching; spores colorless, even, 5-7x4-52 /z.
Fructification 2 cm. high; pileus 1 cm. broad, 13 mm. long;
stem 7 mm. long, § mm. thick, enlarging to 1 mm. where joining
the pileus.
In moss. Labrador. August 8, 1908.
The above description is based on the single dried specimen
collected by the Bryant Labrador Expedition. The small size,
regular obconic form, and very pale color of the membranaceous
pileus and the slender stem are characters making C. horcalis
clearly distinct from other species of Craterellus.
Specimens examined:
Labrador: Gready Harbor, Gready Island, Owen Bryant, type
(in Farlow Herb.).
CYPHELLA
Cyphella Fries, Syst. Myc. 2: 201. 1823.
Fructifications somewhat membranaceous, cup-shaped, rarely
plane, adnate behind, commonly extended in stem-like form,
pendulous; hymenium typically concave or disk-shaped, defi-
nitely inferior in the pendulous species, even or at length rugu-
lose; basidia typically four-spored ; spores subovate or globose,
hyaline, rarely colored.
C digitalis Fries is the type species of this genus.
The fructifications of all our North American species are com-
paratively small, ranging in diameter from a fraction of a milli-
meter for some species to five to fifteen millimeters for those
of the largest species. The fructifications are produced on
the bark of small rotting twigs on the ground and on dead herb-
age, and can only be distinguished from small Pezizce by dem-
onstrating basidia rather than asci in the hymenium. This
demonstration is simply made by crushing under a cover glass
a portion of a fructification in water containing a little seven
per cent solution of potassium hydrate, and then examining
the preparation with the compound microscope. The basidia
are usually four-spored ; in a few species I have as yet been able
to detect only two-spored basidia.
Cyphella is closely related to Solenia by such species as C.
fasciculata and C. mellea, but is separated from it in such cases
1914}
BURT — THELEPHORACE^ OF NORTH AMERICA. Ill 359
by the absence of a hyphal subiculum over the area on which
the fructifications are distributed, and by the less cylindric
form of fructifications of CypJiella. Cyphella is allied to Meru-
lius by C. muscigena and also to Craterellus by this species, speci-
mens of which were described as a Craterellus.
A few species of Cyphella are common and widely distributed,
but most of our North American species are apparently ex-
tremely local and are known only from their respective type
collections. The lack of specimens available for carrying
about to compare with types has been a serious disadvantage
in my study of this genus. Basidia and basidiospores have not
as yet been found for some species which, although originally
referred to Cyphella, have to be regarded as even doubtful Basi-
diomycetes. I have supplemented the original descriptions with
measurements of dried fructifications and with such data in
regard to basidia and spores as the specimens afford. In the
case of very scanty types, the few fructifications are too precious
for gross comparison to be used for microscopic study. For
such species, it seems to me that the descriptions should stand
on the original data, without prejudice, until new collections
become available. Such imperfectly known and partially de-
scribed species are grouped together under the heading ''Species
Imperfectly Known/' Cyphella convoluta Cke., C. Cupressi
(Schw.) Fries and C. suhcyanea Ell. & Ev. are excluded species.
Key to the Species
Fructifications sulphur-colored; hymenium even; spores 4^x2 §-3 fi LC. sulphurea
Fructifications sulphur-colored; hymenium minutely pitted; spores 6-8 x
3-4 M. £. C, Ma
Fructifications white or whitish; on mosses 1
Fructifications white; not on mosses 2
Fructifications neither white nor sulphur-colored 3
1. Fructifications helmet-shaped; hymenium slightly wrinkled; spores 10 x
8 M 5. C. galeaia
1. Fructifications flattened, irregular In form, sometimes stipitate; spores 3-5
X 2-3 /x 4. C. muscigena
1. Fructifications seated upon or developing from webby strings of mycehum
6. C. arachnoidea
2. Fructifications villose, not easily crushed, with a firm base or a short
stem; spores 12-18 x 6-6^ fx 6. C. Tilice
2. Fructifications villose, easily crushed, sessile; spores 10-12x5-7 ^
■ *
7. C villosa
2. Fructifications whitish, minutely webby-hairy, easily crushed, sessile;
spores 8-13 x 4 m 8. C
• *
cancma
[Voiu I
360 ANNALS OF THE MISSOURI BOTANICAL GARDEN
2. Fructifications glabrous, with an oblique stem; spores 4^-^x3-31 m
9. C. capula
2. Fructifications villose, snow-white, sessile, very minute and delicate;
spores 5-6 x 4-4 J fx] from New England 10, C. minutissima
2. Compare with C. cinereo-fasca, C. Palmarum, C. Peckii, C. percxigua, C. pezi-
zoides and C. trachychccla of "Species Imperfectly Known."
3. Fructifications wholly pale ivory-yellow, downy-pubescent, cup-shaped, ses-
sile; spores 4-7 X 3-4 M • ^^- ^' Langloisii
3. Fructifications wholly cream-color, not hairy, helmet-shaped, sessile, re-
supinate-reflexed; hymenium wrinkled; spores 7^x41 fx; on prickle-bear-
ing stems, Jamaica ^^* C^' pomgens
3. Fructifications mineral-gray, tomentose, cup-shaped, sessile; hymenium fus-
cous; spores angular, 4^-6 x4i ^x] on Juniperus IS. C. capidieformis
3. Fructifications wholly gray-pallid, flocculose, sessile; spores 4 x 3 ^
14- C. griseo-jxillida
3. Fructifications externally cinereous, farinaceous, flattened, sessile; hymen-
ium convex, brown; spores 8 x 3J m; on Alnus 15. C. subgelaiino sa
3. Fructifications darker colored than the above 4
4. Fructifications vinaceous-buff, hairy, sessile, f mm. broad; spores 10-12
x 6-8 fx; on bark of Carya 10^ C, Ravenelii
4. Fructifications drying Isabella-color, hairy, sessile, 1-lJ mm. broad;
spores 13 X 8 ju; on Qaercus 17. C, iexensis
4. Fructifications Isabella-color, hau-y, sessile, f-J mm. broad; some spores
colored, 5-6 x 4-4^ ix; on Salix 18. C. mellea
4. Fructifications tawny-oHvo, tomentose, stipitate; often cespitose; spores
7-9 X 2-2| fx; usually on Alnus : 10. C. fasciculata
4. Fructifications fuscous when moist, drying mouse-gray, cespitose and ses-
sile on a common short trunk, glabrous, structure gelatinous 20. C. conglobata
4. Fructifications sepia or olive-brown, cup-shaped, probably glabrous, ses-
sile or with a very short stem; spores 6-8 x 3^-4 /*; on rotting leaves of
Gladiolus ^^. C- fumosa
4. Compare C. BanancB, C. filicicola and C. miisoecola in "Species Imperfectly
Known.''
I.
Iphurea Batsch ex Fries, Hym. Eur. 665. 1874
Peziza sulphurea Batsch, Elenchus Fung. Contin. i! 209.
pi 21. /. 11^6. 1786.— P. campanula Nees, System d. Pilze 268.
/. 295. 1816. — Cyphella sulphurea Batsch, in Patouillard, Tab.
Anal. Fung. 114. /. 266. 1883; Peck, Rep, N. Y. State Mus.
31: 38. 1879.
Illustrations: Batsch, Elenchus Fung. Contin. pi 27. f. 1^6.
—Nees, System d. Pilze /. ;^55.— Patouillard, Tab. Anal. Fung.
^^^.— Oudemans, Ned. Kruidk. Archief XXL 2: pi 3. f. 1-5.
Fructifications scattered or gregarious, membranaceous,
broadly campanulate, somewhat irregular, extended into a short
stem, even, glabrous, sulphur-yellow, the margin somewhat re-
pand; hymenium even; basidia cylindric, 16 x 4J /i>4-spored;
t
1914]
BURT — THELEPHORACEiE OF NORTH AMERICA. Ill 361
spores colorless, even, broadly ovoid, somewhat flattened on
one side, 4^ x 2|-3 n.
Fructifications about 2-3 mm. high; pileus 1-2 mm. broad;
stem 1 mm. long, | mm. thick.
On living stems of herbs in damp places. New York. Sep-
tember. Rare.
The minimum dimensions given above for the fructifications
are about those of European specimens of this species as figured;
the American specimens run rather larger in Peck's collection.
Peck noted that some of his specimens were white when col-
lected, but that they dried yellow like the others of the collec-
tion. In other respects our American specimens agree closely
with the figures and description of European specimens. Oude-
mans gives the spore dimensions as 10-12 x 4-5 n, but Patouil-
lard gives them as they are in American specimens.
Specimens examined:
New York: Griffins, Delaware Co., C. H. Peck (in Coll. N. Y.
State).
2. C. lata Fries, Epicr. 568. 1836-1838.
Illustrations: Patouillard, Tab. Anal. Fung./. 362.
Fructifications membranaceous, obliquely cup-shaped, ex-
tended at the vertex into a stem, pendulous, entire, everywhere
glabrous and sulphur-colored ; stem straight or somewhat flexu-
ous, hymenium minutely pitted; spores colorless, even,
3-4 ju, borne four to a basidium.
Fructifications 3-5 mm. high, 2-4 mm. broad; stem 1-2 mm.
long, about ^ mm. thick.
On dead stems of large herbs lying on the ground. New York.
August .
Fries described the fructifications as 6-8 mm. broad; the
dimensions given above are those of Patouillard 's figures
and of the specimens collected by Peck. Patouillard notes
that the specimens blacken when old; Peck states, ''The
beautiful sulphur-color is lost in drying." The pitted
surface of the hymenium is a noteworthy character of C. Iceta
and this and the larger spores of C. Iceta distinguish it from C.
sulphurea.
Specimens examined:
New York: East Berne, C. H. Peck (in Coll. N. Y. State).
6-8
362 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. I
3. C. galeata Schuin. ex Fries, Epicr. 567. 1836-1838.
Plate 19. fig. 2
Merulius galeatus Schum. Plant. Saellandise 2: 371. 1803
Cantharellus galeatus Fries, Syst. Myc. i : 524. 1821; Flor. Dan.
12: fasc. 34. 11. pi 2027. f. 1. 1830.
Illustrations: Flor. Dan. pi. 2027. f. 1.
Fructifications membranaceous-soft, somewhat sessile, ob-
versely cup-shaped and then dimidiate, helmet-shaped, even,
whitish, the margin entire; hymenium at length rufescent,
slightly \\Tinkled; spores ovate or obovate, 10 x 8 m-
Fructifications 4-15 mm. in diameter.
On mosses. Ohio.
When young entire, cup-shaped; gray when moist, snow-
white when dry, then rufescent. The above description is that
given in European works. The species has been reported from
Ohio by Morgan but I have not studied his specimens nor any
European specimens of this species. The form and coloration
of the pilous and the large spores should distinguish C. galeata
from the other species which occur on mosses in North America.
4. C. muscigena Pers. ex Fries, Epicr. 567. 1836-1838.
Plate 19. fig. 3.
Thelcphora muscigena Pers. Syn. Fung. 572. 1801; Fries, Syst.
Myc. 1 :524. 1821.— T. vulgaris a Candida Pers. Myc. Eur. i :115.
/. 6. 1822. —Cantharellus Icevis Fries, Syst. Myc. i :524. 1821
Elenchus Fung. 55. 1S2S. —Craterellus Pogonaii Peck, Bull. Torr.
Bot. Club 33:218. 1906.
Illustrations: Persoon, Myc. Eur. i : pi. 7.f. ^.— Patouillard,
Tab. Anal. Fung. /. 4^5.— Oudemans, Ned. ICruidk.' Archief
III. 2: pi 11. f. 2.
Pileus mcmbranaceci
ttached by
upper surface, irregular, flattened, white, externally minutely
tomentulose or silky under a lens; stem when present lateral
or eccentric, slender, white; hymenium even or sometimes
rugulose, drying pinkish buff; spores white in collection on slide,
even, apiculate at base, flattened on one side, 4^-5 x 2^-3 n but
only 3-4| x 2-3 n in preparations of the hymenium, borne four
to a basidium.
Pileus 2-6 mm. in diameter; stem when present 3-5 mm.
long, \ mm. thick.
1914]
BURT — THELEPHORACEiE OF NORTH AMERICA. Ill 363
On Polytrichum and other mosses. New England and New
York. August and September.
The fructifications are very variable in form and they are
attached in various ways to the moss plants; they may be some-
what incrusting but at some distance above the ground. The
substance of the pileus is very soft and its upper surface is some-
what bibulous and shows its interwoven fibers under a lens.
The spores of this species are given in Saccardo's 'Sylloge' as
8-10 X 5 /x, but the European specimens of exsiccati cited below
have small spores of the dimensions which I give for American
specimens, and Bresadola, Ann. Myc. i: HI. 1903, gives the
spore dimensions as 3-4 x 3 ju. The specimens of C. Pogonati
were described as sterile by Peck; I find them to be rather im-
mature but bearing spores 3x2/1.
Specimens examined:
Exsiccati: Karsten, Fung. Fenn., 441; Krieger, Fung. Sax., 1564.
Finland: Karsten (in Herb. Fries), and Fung. Fenn., 441.
Germany: Saxony, W. Krieger, Krieger, Fung. Sax., 1564.
Vermont: near Falls of Lana, Salisbury, E. A. Burt.
Connecticut: South Windsor, C, C. Hanmer, 1956, the type
collection of Craterellus Pogonati Pk.
New York: Floodwood, E. A. Burt.
5. C. arachnoidea Peck, Rep. N. Y. State Mus. 44: 134
(22). 1891.
Type : in Collection New York State.
Fructifications membranaceous, very thin, tender, white,
externally downy, irregularly cup-shaped; hymenium some-
what uneven in large specimens; spores colorless, even, some-
what flattened on one side, 4-5 x 3|-4 m, borne at least two to a
basidium.
Fructifications 2-4 mm. in diameter.
On bark and mosses. Vermont and New York. September.
The cups are seated upon or developing from fine, white,
loosely branching, webby strings of mycelium. This is a marked
character in the type and is the chief character for separating
this species from C. muscigena. The spores are slightly more
globose than in the latter and it may be that the hymenium of
C. arachnoidea is superior; in C. muscigena it is inferior. The
hyphse are about 2 ju in diameter in each species.
(Vol. 1
364 ANNALS OP THE MISSOURI BOTANICAL GARDEN
Specimens examined:
Vermont: South Lincoln Notch, near Middlebury, E. A. Burt.
New York: Carrollton, C. II. Peck, type (in Coll. N. Y. State).
6. C. Tiliae Peck ex Cooke, Grevillea 20: 9. 1891.
Plate 19. fig. 16.
Peziza Tilim Peck, Rep. N. Y. State Mus. 24: 96. 1872.
Trichopeziza Tilioe (Peck) Sacc. Syll. Fung. 8: 428. 1889;
Seaver, Proc. Iowa Acad. Sci. 12: 116. 1905; Mycologia i:
110. 1909.
Type : in Collection New York State and a portion from it in
Kew Herbarium.
Fructifications gregarious, rather fleshy, minute, sessile or
nearly so but with firm base, white, globose, then expanded and
concave, drying cup-shaped, densely white villose; hairs straight,
cylindric, granular incrusted, 200 x 6 m; hymenium concave,
even, ivory-yellow to vinaceous buff; spores white in a collec-
tion on a slide, simple, even, ovate, somewhat curved, 12-18 x
6-6 1 fjL, borne four to a basidium.
Fructifications f-1 mm. high, ^-1 mm. broad; stem, when
present, about one-half the height of the whole fructification.
On bark of dead branches of Tilia Americana and Ulmus on
the ground. Canada and Vermont westward to Missouri.
March to October. Probably common.
C. Tilice has somewhat the habit of C. alho-violascens but
differs from the latter in having no violaceous tints, in being
more hairy, in having slenderer spores, and in having at the
base a very firm tubercle which offers considerable resistance
when the fructification is crushed under a cover glass or sec-
tioned. While not cespitose the fructifications of C. Tilice are
often so near together that seven or eight have been counted on
an area a centimeter square. I refer to C. Tilioe many American
specimens which have been distributed under the name C. pezi-
zoides Zopf. The European specimens which Sydow has dis-
tributed under the latter name seem to me from the studies and
comparisons which I made in Kew Herbarium to be C. Curreyi
B. & Br. rather than C. Tilice.
Specimens examined:
Exsiccati: Shear, N. Y. Fungi, 55; Ell. & Ev., N. Am. Fungi,
2316a, under the name C. pezizoides; Ell. & Ev., Fung.
r
V
1D14]
BURT — THELEPHORACE^ OF NORTH AMERICA. Ill 365
Col., 5, under the name C. pezizoides; Rabenhorst, Fung.
Eur., 3942, under the name C. pezizoides.
Quebec: Hull, J. Macoun, 672.
Ontario: Ottawa, J. Macoun, 318, 430; London, J. Dearness,
Ell. & Ev., N. Am. Fungi, 2316a, and Fung. Col., 5.
Vermont: Middlebury, C. 0. Smith, and also E. A. Burt.
New York: Knowersville (Altamont), C. H. Peck, type (portion
in Kew Herb.); Alcove, C. L. Shear, Shear, N. Y. Fungi, 55.
Ohio: Oberlin, F. D. Kelsey (in Mo. Bot. Card. Herb., 4942).
Michigan: Agricultural College, G. H. Hicks, comm. by W. G.
Farlow, 6 (in Mo. Bot. Card. Herb., 43807).
Wisconsin: Blue Mounds, I. E. Melhus, comm. by C. J. Hum-
phrey, 2410 (in Mo. Bot. Card. Herb.).
Missouri: C. H. Demetrio, Rabenhorst, Fung. Eur., 3942.
7. C. villosa Pers. ex Karsten, (Mycol. Fenn. 3) Bidrag Finska
Vet.-Soc. 25 : 325. 1876. Plate 19. fig. 13.
Peziza villosa Pers. Syn. Fung. 655. 1801; Fries, Syst. Myc.
2: 104, pr. p. 1823. — An Cyphella pezizoides Zopf, in Morgan,
(Myc. Fl. Miami Val.) Jour. Cincinnati Soc. Nat. Hist. 10:
202. 1888?
Illustrations: Patouillard, Tab. Anal. Fung./. 257.
Fructifications gregarious, membranaceous, sessile, drying
globose or obconic and with the pore nearly closed by the hairs,
white, externally white-villose; the hairs granular incrusted,
cylindric, 200 x 5-6 n; hymenium even, concave; spores hya-
line, even, ovoid, flattened on one side, broadest near the base,
10-12 x 5-7 ju.
Fructifications about I mm. high, ^-j mm. broad.
On dead stems of Artemisia, Helianthus, and Solidago. South
Carolina, Missouri and California. June and July.
The fructifications of C. villosa resemble those of C. Tilice in
form, color, and hairiness but are much smaller than those of
C. Tilice, more membranaceous and easily crushed under a cover
glass, and have smaller spores. The hymenium is very pale
with not more than a very slight yellowish tint.
Specimens examined:
Exsiccati: Krieger, Fung. Sax., 1457; Ravenel, Fung. Am.,
459; Ell. & Ev., N. Am. Fungi, 2316b, under the name
Cyphella pezizoides Zopf.
[Vol. 1
366 ANNALS OF THE MISSOURI BOTANICAL GARDEN
South Carolina: Aiken, Ravenel, Ravenel, Fung. Am., 459.
Missouri: Emma, C. H. Demetrio, Ell. & Ev., N. Am. Fungi,
2316b.
California: Half-moon Bay, San Mateo Co., E. B. Copeland,
Baker, Pacific Coast Fungi, 3611 (in Mo. Bot. Gard, Herb.,
4944).
8. C. caricina Peck, Rep. N. Y. State Mus. 33: 22. 1880.
Plate 19. fig. 8.
Type: in Collection New York State.
Fructifications scattered, membranaceous, sessile, wholly
white, externally minutely webby-hairy; hymenium glabrous,
uneven in large specimens; basidia cylindric, 20 x 5 /x, 4-spored;
spores colorless, even, lanceolate or subclavate, pointed at base,
8-13 X 4 ju.
Fructifications 1-2 mm. broad.
On culms and leaves of carices. New York. August.
The spores of the type are noteworthy by their tapering base.
Specimens examined:
New York: Verona, C. H. Peck, type (in Coll. N. Y. State).
9. C. capula Holmsk. ex Fries, Epicr. 568. 1836-1838.
Plate 19. fig. 4.
Peziza Capula Holmsk. Nov. Act. Havn. i : 286. /. 7; Fung.
Dan. 2:41. pi. 22. 1899.
Illustrations: Holmskiold, Nov. Act. Havn. i : 286./. 7; Fung.
Dan. 2: pi. 22.—Y\g^. Dan. 33: pi. 1970. f. 3.— Patouillard, Tab.
Anal. Fung, i : /. 35.
Fructifications membranaceous, obliquely campanulate, ex-
tended into an oblique stem, glabrous, whitish, the margin sin-
uate, irregularly shaped; hymenium even. ... On dead
stems of herbaceous plants.
— Translation of description in Fries' 'Epicrisis.'
Fructifications in the figures of Holmskiold 4-9
mm
pileus 2-7 mm. long, 2-4 mm. broad; stem 1-2 mm. long.
On dead stems of Fa^mculum and other herbs. New York
and South Carolina.
I have not been able to study any European specimens of
species. In the copv of Cooke's 'Fungi Britannici' in the
this species. In the copy
herbarium of the Missouri Botanical Garden the packet labeled
C. capula, 1 12, contains onlv some nieces of stubble. The Amer-
1014]
BURT — THELEPHORACE^ OF NORTH AMERICA. Ill 367
ican specimens distributed in Ravenel's 'Fungi Americani,'
458, were determined by Cooke. In their present dried condi-
tion these specimens agree well with Holmskiold's illustrations
in form; the stem of these specimens is now hair-brown and the
pileus pale olive-buff; their dimensions are: fructifications 1-3
mm. long, pileus 1-2 mm. long and broad; stem |-1 mm. long
X 100 IX thick. The basidia are 16-20 x 3^-4^ n; spores color-
less, even, flattened on one side, 4^-6 x 3-3 1 (i.
Specimens examined:
Exsiccati: Ravenel, Fung. Am., 458.
South Carohna: Aiken, Ravenel, Ravenel, Fung. Am., 458.
10. C. minutissima Burt, n. sp. Plate 19. fig. 5.
Type: in Mo. Bot. Gard. Herb, and in Farlow Herb.
Fructifications gregarious, very minute, membranaceous and
very delicate, sessile, globose, snow-white, externally villose,
often with mouth oblique, margin inrolled; hairs white, in-
crusted, 75-90 X 4 /x; hymenium concave, white; basidia cla-
vate, 16x4 /x; spores colorless, even, 5-6 x 4-4|jli.
Fructifications 200-500 m broad, about 200-500 ix high.
On inner bark of Populus. New Hampshire. August.
The characters of this species agree in some details with those
in the incomplete description of C. glohosa Pat., the specimens
of which were collected on the under side of leaves of ferns in
Ecuador by von Lagerheim, but as no mention is made of spore
characters for C glohosa and as other species of Cyphella have
not been found to vary widely with regard to kind of substra-
tum, it seems best to regard our New England species as proba-
bly distinct. C. punctiformis (Fries) Karst. is a small white
Cyphella, described by Karsten as having spores 5-8x2-4 n]
I have not been able to study authentic specimens of C. punc-
tiformis, but comparison of C. minutissima with this species of
northern Europe should be made.
I refer to C. minutissima a collection made by myself in Ver-
mont on bark of rotting locust limbs. The fructifications of this
collection lack spores but agree in all other respects with the
type.
Specimens examined:
New Hampshire: Chocorua, W. G. Farlow, 3, type (in Mo.
Bot. Gard. Herb., 43803, and in Farlow Herb.).
Vermont: Middlebury, E. A. Burt.
[Vol. 1
368 ANNALS OF THE MISSOURI BOTANICAL GARDEN
11. C. Langloisii Burt, n. sp. Plate 19. fig. 6.
Type: in Farlow Herb, and Burt Herb.
Fructifications gregarious, membranaceous, cup-shaped, ses-
sile, drying pale ivory-yellow, externally downy pubescent, the
margin inrolled; hairs colorless, somewhat crinkled together,
granular incrusted, 100-150 x 3|-4| /x; hymenium concave, even,
pale ivory-yellow to cream color; spores colorless, even, pointed
at the base, 4-7 x 3-4 fj.; basidia clavate, 20 x 5m, 2-spored.
Fructifications about | mm. high; *-^ mm. broad.
On dead stems of Arundinaria and on decaying pieces of wood
lying on the ground. Louisiana. September and April,
The fructifications of C Langloisii are about as small as those
of C, minutissima but differ from them in being somewhat ex-
tended laterally and occasionally somewhat laterally confluent
rather than always globose, in having an ivory-yellow rather
than snow-white color, and in having the hymenium colored
and the hairs longer than in C. minutissima. Comparison should
be made with C . fraxinicola B. & Br., of which I have studied no
specimens but which seems distinct by some characters of the
incomplete published description.
Specimens examined:
Louisiana: St. Martinville, A. B. Langlois, 1802, type (in Farlow
Herb.), and cz, type, in Burt Herb., and cy, and from the
same collector but comm. by W. G. Farlow, 5 (in Mo.
Bot. Card. Herb., 43791).
12. C. porrigens Burt, n. sp. Plate 19. fig. 7.
Type: in Burt Herb, and New York Bot. Gard. Herb,
Fructifications scattered, membranaceous, thin, wholly cream-
color, sessile, obversely cup-shaped or helmet-shaped, resupinate
by the upper surface of one side but with the greater portion of
the pileus extended and reflexed; hymenium inferior, somewhat
wrinkled when moistened, concave, basidia clavate, 20-25 x
4-4^ /i, with four sterigmata; spores colorless, even, flattened on
one side, obovate, 7-| x 4| n.
Fructifications ^-1 mm. broad.
On dead prickle-bearing stems, possibly Ruhus sp. Wet
mountainous region at altitude 4500-5200 feet. Cinchona,
Jamaica. About January 1.
This species does not appear closely related to any other
BURT — THELEPHORACE^ OF NORTH AMERICA. Ill OD^
species; it is marked by the resupinate-reflexed habit of most
fructifications; only rarely is a fructification attached by its
vertex. The dried specimens are externally minutely fibrillose
under a lens but do not show hairs in microscopic preparations.
When the fructifications are moistened the hymenium shows
two or three minute wrinkles radiating from an eccentric point.
Specimens examined:
Jamaica: Cinchona, W. A. and Edna L. Murrill, N. Y. Bot.
Gard., Fungi of Jamaica, 607, type.
13. C. cupulaeformis Berk. & Rav. Grevillea 2: 5. 1873.
Plate 19. fig. 9.
Type: type and cotype in Kew Herb, and in Curtis Herb.
respectively.
Fructifications scattered, rarely in clusters of two or three,
sessile, cup-shaped, somewhat globose, externally mineral gray
and obscurely tomentose, the margin incurved; hymenium
concave, even, fuscous; basidia clavate, 20-25 x 4-6 n, having
2-4 sterigmata which become finely attenuated ; spores colorless,
angular, 4|-6 x 4| /i.
Fructifications | mm. high, ^-1 mm. broad.
On bark of Juniperus lirginiana. South Carolina and Georgia.
The hairiness of the exterior of the pileus is due to the irregu-
larly curved and interwoven hyphse which form the surface
layer of the pileus ; these hyphaj are colorless and about 3 /x in
diameter, and they bear scattered but large incrusting granules.
The angular spores of this species are often octahedral in form
and are noteworthy for Cyphella; at maturity, they are attached
to the basidium by sterigmata becoming 6 m long and so finely
attenuated that the attachment of the spores to the basidia is
made out with difficulty. This species may be readily known
by its occurrence on bark of Juniperus virginiana and by its
angular spores.
Specimens examined:
Exsiccati: Ravenel, Fung. Am., 224.
South Carolina: Ravenel, U03, type (in Kew. Herb.).
Georgia: Darien, Ravenel, Ravenel, Fung. Am., 224.
14. C. griseo-pallida Weinm. Hymeno- et Gastero-mycetes in
Rossico. 522. 1836.
Illustrations: Patouillard, Tab. Anal. Fung. /. 255.
[Vol. 1
370 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Fructifications gregarious, adnate-sessile, membranaceous,
wholly gray-pallid, externally flocculose; hymcnium glabrous,
even.
At first having the form of globose, closed granules, soon
open, campanulate or crateriform, often dimidiate in old stages.
Fructifications ^ mm. high, ^-2 mm. broad.
On moist ground and on pine wood thinly covered with earth
and on old cracked trunks of Lonicera iartarica (in Europe).
— Translation of original description.
On bark, twigs and leaves lying on the ground. New York
and Ohio. November.
I have not seen the type of C. griseo-pallida nor any European
specimens which have been compared with it, but Peck, Rep.
N. y. State Mus. 30: 48. 1879, has referred to this species a
collection which he made at Sand Lake, New York. Peck notes
that his specimens sometimes have a very short stem. I found
the spores of these specimens hyahne, even, somewhat flattened
on one side, 4 x 3 ju; basidia 12 x 4 /x.
Specimens examined:
New York: Sand Lake, C. H. Peck (in Coll. N. Y. State).
15. C. subgelatinosa Berk. & Rav. Grevillea 2: 5. 1873.
Type: in Kew Herb.
Fructifications scattered, somewhat gelatinous, sessile, flat-
tened, externally cinereous and farinaceous, the thin margin
inflexed; hymenium shghtly convex, even, brown; basidia cla-
vate, about 25 x 5-6 m, probably 2-spored; spores colorless, even,
ellipsoidal, 8 x 3| /i.
Fructifications about U mm. broad.
On Alnus serrulata. South Carolina.
The fructifications of the type have dried with the slightly
convex hymenium so prominently visible that they resemble
brown apothecia of hchens with a pale margin (exciple). The
most of the basidia are immature; I found one showing two sterig-
mata distinctly. No spores were found attached to basidia;
the spore characters, which are given above, are those of loose
spores in the preparation. (7. subgelatinosa is so very distinct
from our other species of Cyphella that it will probably be over-
looked by botanists collecting Basidiomycetes only, unless es-
pecially kept in mind.
1914]
BURT — THELEPHORACEiE OP NORTH AMERICA. Ill 371
Specimens examined :
South Carolina: Aiken, Ravenel, 1714, type (in Kew Herb.)-
i6. C. Ravenelii Berk. Grevillea 2 : 5. 1873. Plate 19. fig. 14.
Type: type and cotype in Kew Herb, and in Curtis Herb.
respectively.
Fructifications single or gregarious, sessile, subglobose, some-
what flattened, depressed at the pore, minutely hairy under a
lens, vinaceous buff; hairs minutely rough, about 300 ^u long,
4 ju thick, tapering towards the free end, olive-yellow under the
microscope; spores hyaline, or perhaps very slightly colored,
even, broadly ellipsoidal, 10-12 x 6-8 ju.
Fructifications 0.6 mm. high, 0.8 mm. broad; pore 0.15 mm.
in diameter.
On bark of Carya. South Carolina.
The specimens of this species which I have seen have been on
thick and cracked portions of bark apparently from large
branches or the main trunk of the tree. Sometimes only one
fructification occurs on a piece of bark a centimeter square;
sometimes such a piece bears from 3 to 6 fructifications with
some of them barely in contact with one another. The type
specimen contains so few fructifications that I made a micro-
scopic preparation at Kew Herbarium from the specimen dis-
tributed by Ravenel in Ellis, N. Am. Fungi, 721, which seems
to me to be certainly the same species as the type. Berkeley
described the spores in his original description as ''elliptic,
.00025 (in.) long"; I found them about twice this length in my
preparation referred to and also in a preparation recently made
from the specimen in Ravenel, Fung. Am., 130, in the Mo. Bot.
Gard. Herb.
Specimens examined:
Exsiccati: Ravenel, Fung. Am., 130; Ellis, N. Am. Fungi, 721.
South Carolina: Aiken, Ravenel, 1755, the type and cotype (in
Kew Herb, and in Curtis Herb, respectively); and also
Aiken, Ravenel, Ravenel, Fung. Am., 130, and Ellis, N. Am.
Fungi, 721.
17. C. texensis Berk. & Curtis, Grevillea 20: 9. 1891.
Plate 19. fig. 10.
Type: in Kew Herb.
Fructifications scattered, sessile, pallid but at present time
372 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. I
Isabella-color (melleus of 'Chromotaxia'), cup-shaped, at length
flattened and disk-shaped, externally hairy; hairs olive-ocher
under the microscope, granular incrusted, cylindric, 300-400
X 4^-G n; basidia clavate, 25-30 x 6-8 n, 4-spored; spores hyaline,
even, broadly ellipsoidal, 13 x 8 ju.
Fructifications 1-1^ mm. broad.
On Quercus. Texas.
The type is scanty, consisting of three fructifications, but
these fructifications are in fine condition and present well the
characters of the species. C. texensis now impresses me as more
closely related to C. Ravenelii than I observed when studying
the specimens of both in Kew Herbarium. The fructifications
of C. texensis are the melleus of Saccardo's ' Chromotaxia' and
the hairs are of a little greater diameter and have larger incrust-
ing granules than those of C. Ravenelii, but the spores and
basidia are very similar in form and dimensions in both species.
Specimens examined:
Texas: Wright, 3779, type (in Kew Herb.).
i8. C. mellea Burt, n. sp. Plate 19. fig. 12.
Type: in Burt Herb, and in U. S. Dept. Ag. Herb.
Fructifications closely gregarious, sessile, Isabella-color, spher-
ical and with margin inroUed in the dried state, sometimes
obconic, externally hairy; hairs granular incrusted, baryta-
yellow under the microscope, cylindric, 80-100 x 3|-4 n] hy-
menium even, whitish or pale olive-buff; basidia clavate, 12-16
x 6 M) spores mostly colorless but some pale baryta-yellow, even,
broadly ellipsoidal, 5-6 x 4-4^ /x.
Fructifications about J-i mm. high and broad.
On rotten wood of Salix nigra. Louisiana. December.
In the specimen upon which the description is based, the most
of the fructifications are about ^ mm. high and broad and are
distributed on the rotten wood at the rate of about 200 per
square centimeter. Rarely a short stem-like base is visible when
the fructifications emerge from the bottom of small crevices
between the fibers of the wood, but the fructifications are gen-
erally sessile. The species is intermediate between Cyphella
and Solenia but is included in the former genus because the fruc-
tifications do not arise from a common subiculum and are more
globose than in Solenia. The description of C. mellea suggests
1914]
BURT—THELEPHORACE^ OF NORTH AMERICA. Ill 373
those of C. Ravenelii and C. texensis in many respects, but the
fructifications are much smaller and more numerous than in
either of these species, and their various parts are also much
smaller and some of the spores are colored.
Specimens examined:
Louisiana: Bohemia, Plaquemines Co., A. B. Langlois, 864a,
type, in Burt Herb, and also (in U. S. Dept. Ag. Herb.);
A. B. Langlois, 864 (in U. S. Dept. Ag. Herb.).
19. C. fasciculata Schw. ex Berk. & Curtis, Jour. Acad. Nat.
Sci. Phila. 3 : 207. 1856. Plate 19. fig. 17.
Caniharellus fasciculatus Schw. Trans. Am. Phil. Soc. N. S.
4: 153. 1S31 .~~C . fasciculatus Schw. in Saccardo, Syll. Fung. 5:
495. 1S87. —Cyphella fasciculata Berk. & Curtis, Grevillea 2:
6. 1873. — Solenia anomala Pers. var. orhicularis Peck, Rep. N.Y.
State Mus. 47: 168 (42). ISM.—Cyphella fulva Berk. & Rav.
Grevillea 2: 5. 1873.— C. Ravenelii Saccardo, Syll. Fung. 6: 672.
1888.— C. Saccardoi Sydow, in Saccardo, Syll. Fung. 14: 233.
1900.— C. furcata Berk. & Curtis, Grevillea 2 : 5. 1873.
Type: in Herb. Schweinitz.
Fructifications gregarious, sometimes fascicled, pezizoid,
tawny olive; pileus stipitate, cup-shaped, extended vertically or
pendulous, tomentose with tawny-olive, even-walled hairs
which are flexuous or somewhat spirally curved towards the
tips, the margin strongly inrolled; stem short, variable in length,
cylindric, tomentose, colored like the pileus; hymenium concave,
even, drying olive-buff; spores hyaline, even, cylindric, shghtly
curved, 7-9 x 2-2^ n, borne four to a basidium.
Fasciculate clusters about 2 mm. in diameter, 1 mm. high;
fructifications f-l mm. in diameter, 1-2 mm. high; stem §-1
mm. long, 3-i mm. thick.
On bark of twigs of Alnus in swamps and rarely on Prunus
virginiana and Pyrus Malus. Canada and Newfoundland to
South CaroHna and westward to Wisconsin. Throughout the
year, more highly fasciculate from autumn to spring. Common.
This fungus is very common on dead twigs of Alnus in swamps.
The color is similar to that of Solenia anomala but the fructifi-
cations are rather larger and more cup-shaped than those of the
latter and have the hymenium merely concave rather than lining
a tube. The fructifications burst out through the outer bark
2
[Vol. 1
374 ANNALS OF THE MISSOURI BOTANICAL GARDEN
either singly or in clusters of from two to twenty individuals
more or less connected together at the base. The differences in
habit between the extremes of highly fascicled forms and those
with fructifications gregarious and largely single, impress one
as of specific weight at first and I should like to recognize these
extremes as two species but they intergrade too completely.
The dated collections which I have seen, indicate that the speci-
mens become highly fasciculate in autumn and winter.
I do not understand why Berkeley attempted authorship for
this species. The C. fasciculata B. & C. is certainly that of
Schweinitz both in description and in fascicled form of types;
and as for C. fulva B. & Rav., it is noted in the original descrip-
tion that it is the same as Cantharellus fasciculatus Schw.
Specimens examined:
Exsiccati: Ellis, N. Am. Fungi, 936, fascicled form; Ell. & Ev.,
Fung. Col., 1818; fascicled form under the name C. RaveneUi
Berk.; Shear, N. Y. Fungi, 308, fascicled form under the
name Solenia anomala (Pers.) Fr. var. orhicularis. Pk.
Peck det.; Ravenel, Fung. Car. IV., 16, the type distribu-
tion of C. fulva B. & Rav.; Ravenel, Fung. Am., 129 (bear-
ing spores in abundance) ; Shear, N. Y. Fungi, 56.
Newfoundland: Headquarters, B. L. Robinson & H. von Schrenk
(in Mo. Bot. Card. Herb., 4764 and 43789, the latter com-
municated by W. G. Farlow); Bay of Islands, A. C. Wag-
Jiorne, 127 (in Mo. Bot. Card. Herb., 42593).
Quebec: Hull, J. Macoun, S55.
Ontario : Ottawa, J. Macoun, S3.
Maine: /. Blake (in Curtis Herb., 6926, and in Kew Herb.).
New Hampshire: Conway, W. G. Farlow; North Conway, W.
G. Farlow (in Mo. Bot. Card. Herb., 43786); Shelburne,
H. von Schrenk (in Mo. Bot. Card. Herb., 4765), W. G.
Farlow (in Mo. Bot. Card. Herb., 43787); FrankUn Falls,
Mrs. J. B. Harrison, Ellis, N. Am. Fungi, 936.
Vermont: Middlebury, on Alnus and on Prunus virginiana,
E. A. Burt.
Massachusetts: Newton, W. G. Farlow (in Mo. Bot. Card,
Herb., 42591, 42592 and 43788).
New York: Torrey, type (in Herb, ^(ih.-^.) •,Sartw ell, Qoiy^e and
typeof C./asc7CwZatoB.&C. (in Curtis Herb., 2659, and in
1914]
BURT — THELEniOEACE^ OF NORTH AMERICA. Ill 375
Kew Herb, respectively) and specimen (in Mo. Bot. Gard.
Herb., 4937); Ithaca, G. F, Atkinson; East Galway, E. A.
Burt; Keeseville, C. 0. Smith, Ell. & Ev., Fung. Col., 1818;
Alcove, C. L. Shear, Shear, N. Y. Fungi, 56 and 308; Albany,
C. H. Peck, comm. by H. D. House (in Mo. Bot. Gard.
Herb., 43821); Earner, C. H. Peck, comm. by H. D. House
(in Mo. Bot. Gard. Herb., 43820).
South Carolina: Ravenel, 1683 (in Curtis Herb, and in Kew
Herb.), and in Ravenel, Fung. Car. IV., 16; Aiken, Ravenel,
Ravenel, Fung. Am., 129.
Alabama: Beaumont, the cotype and type of C. furcata (in Curtis
Herb., 4022, and in Kew Herb, respectively).
Wisconsin: Madison, W. Trelease (in Mo. Bot. Gard. Herb.,
42594).
20. C. conglobata Burt, n. sp. Plate 19. fig. 15.
Type: in Mo. Bot. Gard. Herb, and in Farlow Herb.
ons cespitose, 10-30 together, sessile on a common
Fructificat
short trunk which is erumpent through the bark; individual
fructifications subglobose, fuscous and glabrous when moist,
drying mouse-gray and with the margin inrolled; hymenium
concave, black or nearly black; basidia simple, with four sterig-
mata; spores colorless, even, cylindric, shghtly curved, 8-10
X 2^-3/1.
Cluster 1-2 mm. in diameter, emerging about ^ mm. from the
bark; cups 400-500 n broad, nearly as high.
Clusters scattered on small limbs of Alnus. New Hampshire
and New York. July and September.
The clusters of this curious fungus are distributed at the rate
of about 5 or 6 clusters to the square centimeter on what I con-
clude to have been the under side of a horizontal limb — perhaps
a limb prostrate on the ground; for cups in clusters exactly on
this presumably under side have the pore central while in the
clusters which emerged more obliquely from the limb the cups
are somewhat auriform with oblique pore and are arranged in
imbricated manner. The outer surface of the cups is composed
of irregularly branched and interwoven pale brownish hyphae
about 2 /x in diameter. The substance of the fructifications
and common trunk-like base is composed of colorless hyphae
with walls gelatinouslv modified.
[Vol. 1
376 ANNALS OF THE MISSOURI BOTANICAL GARDEN
One might regard this fungus as the type species of a new
genus distinct from Cyphella or Solenia by the common central
mass on which the individual cups are borne, but in Cyphella
fasciculaia the cups sometimes occur singly and sometimes
branching from a common central or basal mass . For this reason
it seems best to include the present species in Cyphella through
its relationship in plan of structure to C. fasciculata, from which
it is specifically distinct in other respects, however. Both these
species are excluded from Solenia by their short and globose
fructifications and by the absence of a subiculum on the general
area over which the clustered fructifications are distributed.
Specimens examined:
New Hampshire: Lower Bartlett, R. Thaxter, comm. by W. G.
Farlow, 4, type (in Mo. Bot. Gard. Herb., 43806, and in
Farlow Herb.).
New York : Adirondack Mts., C. H. Peck, comm. by H. D. House
(in Coll. N. Y. State and in Mo. Bot. Gard. Herb., 43818);
North Elba, C. H, Peck, comm. by H. D. House (in Mo.
Bot. Gard. Herb., 43819).
ai.
fumosa Cooke, Grevillea 20: 9. 1891. Plate 19. fig
Type: in Kew Herb.
Fructifications gregarious, membranaceous, cup-shaped,
flcxuous, sepia or olive-brown and blackening, even, attenuated
below into a very short stipe, or sessile; hymenium even; basidia
cylindric-clavate, 20 x 4-5 m; spores colorless, even, somewhat
flattened on one side, 6-8 x 3^-4 /x.
Fructifications 1-2 mm. broad.
On rotting leaves of Gladiolus. South Carolina.
Cooke described the spores of this species as globose, 4 m in
diameter, but I found no such spores in my preparation from
the type. Spores 6-8 x 3W m are abundant and are probably
the spores of this species, although I could not find any spores
still attached to the basidia. I conclude from my microscopical
preparations that the fructifications are glabrous.
Specimens examined:
South Carolina: Aiken, Ravenel, 3071, type (in Kew Herb.).
1914]
BURT — THELEPHORACE^ OF NORTH AMERICA. Ill 377
SPECIES IMPERFECTLY KNOWN
C. cinereo-fusca Schw. ex Saccardo, Michelia 2: 303. 1881.
Peziza cinereo-fusca Schw. Schrift. d. Naturforsch. GeselL,
Leipzig, i: 119. 1822; Fries, Syst. Myc. 2: 97. 1823.— Ct/-
phella cinereo-fusca (Schw.) Sacc. Syll. Fung. 5: 674. 1888.
Lachnella cinereo-fusca (Schw.) Sacc. Syll. Fung. 8: 399.
1889.
Fructifications minute, gregarious, sessile, externally fari-
naceous-hirsute and ash-green, the margin incurved ; hymenium
fuscous-bay.
On decorticated branches of Cercis. [North Carolina.]
3 mm. broad. Cups often closed.
— Translation of original description.
I have not seen an authentic specimen of this species nor any-
thing on Cercis which seems referable to it. The species is
given here on the authority of Saccardo, /. c, who refers to this
species a Cyphella collected on Vitis vinifera near Toulouse,
France, by Roumeguere. Saccardo does not state that he
made comparison with an authentic specimen from Schweinitz,
and he has entered the species in the 'Sylloge Fungorum' in
both the B asidiomy cetes and the Discomycetes.
C. Palmarum Berk. & Curtis, (Fung. Cub.) Jour. Linn. Soc.
Bot. 10: 337. 1867.
Type: type and cotype probably in Kew Herb, and Curtis
Herb, respectively.
White, pileus cyathiform, externally obscurely pruinose; stem
short, tomentose, rather thick.
Scarcely 2 mm. high; stem rather thick for the size of the
pileus, often oblique.
On petioles of palms. Cuba. June. C. Wright, 753.
;ed from original descript
Sacc. Syll. Fung. 6
C. Candida Peck, Rep. N. Y. State Mus. 27: 99. 1875.
Type: in Coll. N. Y. State.
Fructifications scattered or gregarious, membranaceous, soft,
obconic, nearly or quite sessile, sometimes deflexed, wholly
white, externally tomentose; hairs tapering to a sharp point,
roueh -walled, 60-70 x 31 j".
378 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
Fructifications about 1 mm. broad.
On dead stems of ferns, Osmunda cinnamomea. New York.
September.
The type specimens of this species are immature. I could
make out neither distinct asci nor basidia in the hymenium.
In a crushed preparation I found one spore, colorless, even,
pointed
It may have been a basidio
whitish
of this species or it may have been a foreign spore.
Specimens examined:
New York: Forestburgh, C. H. Peck, type (in Coll. N. Y,
State).
C. perexigua Sacc. Michelia 2: 136. 1880.
Cups bell-shaped, very short and obliquely stipitate, small,
L. long, thin-membranaceous, internally and externally
cinereous, externally minutely puberulent; spores not
seen. Appears related to C. erucceformis and cupuliformis but
is one-third as large. ... On decorticated branches.
South CaroHna. Ravenel. — Translation of original description.
I have not seen the type of C. 'perexigua, which is probably in
Saccardo Herb. As basidia and basidiospores have not been
found for American specimens, it is uncertain whether this
species is a Cyphella. Patouillard, Tab. Anal. Fung. 19. /. 34.
1883, referred to C. perexigua a species of Cyphella which he
collected at Poligny, France, but that reference is doubtful in
the absence of knowledge in regard to basidia and basidiospores
for American specimens.
C. pezizoides Zopf, in Morgan, (Myc. Fl. Miami Val.) Jour.
Cincinnati Soc. Nat. Hist. 10: 202. 1888.
Type: probably in the State Univ. of Iowa Herb.
"Fructifications membranaceous, nearly sessile, globose then
cup-shaped, clothed externally with long erect white hairs.
Hymenium even, brownish; spores obovate, .012-.013
length.
"On old herbaceous stems; not common, cupule pezizoid
scarcely pedicillate, about half a line in diameter. The lonj
hairs are erect and connivent over the hymenium; they are hya^
mm
and incrusted with crvstals of calcium
The type is not accessible at present.
Original description.
BURT — TIIELEPHORACEiE OF NORTH AMERICA. Ill 379
C. trachychseta Ell. & Ev. Jour. Myc. 4:73. 1888.
Type: in New York Bot. Gard. Herb.
Fructifications gregarious, sessile by a narrow base, white,
cup-shaped, clothed outside with appressed hairs; hairs subhy-
aline, very rough, with a smooth tapering tip 12-15 m long; hairs
paler around the base of the fructification and coarsely roughened
by irregularly shaped tubercles, some of which are prolonged
into short spines; hymenium nearly white with a slight tinge of
slate color; basidia and spores could not be well made out, but
the latter are apparently very minute.
Fructifications 300-400^ high and broad, occasionally 1 mm.
broad and with the margin distinctly lobed.
On fallen leaves of Quercus. Louisiana. July.
The above description is arranged from that originally pub-
lished. I am under obligation to Dr. Murrill for recently send-
ing to me a portion of the type for study, but the specimen
proves too immature to show whether this species is a basidio-
mycete. The hymenium of this specimen is now pale olive-
buff; the hairs are 50-75 x 6 /i, heavily encrusted except near the
tips, but I failed to find any hairs roughened by tubercles or
bearing spines.
Specimens examined:
Louisiana: il. B. Langlois, I424, type (in N. Y. Bot. Gard.Herb.).
C. Bananae Cooke, Grevillea 6: 132. 1878.
Type: probably in Kew Herb.
Fructifications fuliginous or wood-brown, finger-shaped,
pendulous-extended behind, glabrous, the margin entire; hy-
menium white, rugose; spores linear, obtuse, curved, 10-12 x
2h n
of original descript
On dead leaves of Musa. Gainesville, Florida. Ravenel
C. filicicola Berk. & Curtis, Grevillea 2:5. 1873.
Type: type and cotype probably in Kew Herb, and Curtis
Herb, respectively.
Stem very short; cups irregular, sometimes oblique, externally
very obscurely tomentose, umber.
On dead fern. North Carolina. Curtis Herb., 4934, type.
The above contains all the items of the original description;^!
overlooked this species when studying in Curtis Herb, and in
Kew Herb.
380
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
C. musaecola Berk. & Curtis, Jour. Linn. Soc. Bot. lo: 337.
1867.
Type: type and cotype in Kew Herb, and Curtis Herb,
respectively.
Pileus crucible-form, pallid purple, with very short stem or
sessile, externally tomentose; hymenium luteus (cadmium-
y^^^ow). —Translation of original description.
About 2 mm. across.
On sheaths of plantain leaves. Cuba. C. Wright, 75 L
By the kindness of Dr. Farlow I have been permitted to
examine a specimen from the type collection. I fail to find any
fructifications of a Cyphella present. A leaf-spot fungus has
caused some dark purple discolcrations 1-2 mm. in diameter at
Specimens
surface of the leaf
Cuba: C. Wright, 751, comm. by W. G. Farlow (in Mo. Bot
Card. Herb
EXCLUDED SPECIES
C. convoluta Cooke, (Fungi of Texas) Ann. N. Y. Acad. Sci
1 : 179. 1878.
Type: In Kew Herb.
"Scattered, cup-shaped, then flattened, 1 to 2 mm. wide
margin membranaceous, involute, externally \
fleshy-red; spores oblong (.007 mm. long).
hit
''On trunks. Ravenel (295)."— The original description.
I examined the type of this fungus, which was collected at
Houston,^ Texas, and do not regard it as a Cyphella. The
''basidia" are filiform and only 1-spored; spores are abundant,
hyaline, even, 4-5 x 2-2^ n.
C. Cupressi Schw. ex Fries, Epicr. 567. 1836-1838.
Merulius Cupressi Schweinitz, Schrift d. Naturforsch. Gesell
Leipzig, i: 92. 1822. '
This species is an insect gall, not a Basidiomycete. Its true
nature seems to have been first pointed out by Berkeley &
Curtis, Jour. Acad. Nat. Sci. Phila. 3: 207. 1856.
C. subcyanea EII. & Ev. Jour. Myc. 2: 37. 1885.
As this species is not mentioned in Saccardo's 'Sylloge Fun-
gorum' and as the early numbers of the Journal of Mycology are
pare, I quote the original description as follows :
1914]
BURT — THELEPHORACEiE OF NORTH AMERICA. Ill 381
"On living leaves of Sabal Palmetto, Louisiana, Nov. 1885.
Rev. A. B. Langlois, No. 57. Shallow cup-shaped, thin, substi-
pitate, oblique, less than 1 mm. across, whitish and nearly
smooth outside, hymenium bluish or lead colored. Spores
filiform multinucleate, upper end thickened, curved into a
semicircle, 40-60 m long by 1§ /z thick, on short (11-12 x l|-2 ^
subcylindrical sporophores, which are a little thickened below."
This species was distributed in 1891 in Ell. & Ev., N. Am.
Fungi, 2602, the specimens having been collected on living
stems of Smilax in Louisiana by Mr. Langlois. Mr. Langlois
communicated to me still better specimens on dead canes of
Arundinaria. The fructifications occur scattered here and there
in grayish areas 2-4 mm. long by ^-1 mm. broad on the surface
of the stems. Dr. Farlow informs me in a letter as the proofs
are at hand that the above species is the lichen Heterothecium
Augustinii Tuckm.
(To be continued.)
[Vol, 1, 1914
382 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 19
The figures of this phxle have been reproduced natural size from photo-
graphs of dried herbarium Hi)ecimens except in the cases noted otherwise.
Fig. 1. Cralerdlus horealis. From the type specimen collected at Gready Island,
Labrador, by Owen Bryant.
Fig. 2. Cyphella galeata. From photograph, natural size, of the figure hi Flor.
Dan. pi S027,f. L
Fig. 3. C. muscigcna. The two figures on the left are from specimens collected
at Floodwood, New York, by E. A. Burt; the two on the right are from the type
collection of Craterellus Pogonaii collected at South Windsor, Connecticut, by C C.
Hanmer, 1956.
Fig. 4. C capula. From photograph, natural size, of the figure in Fung. Dan.
2 : pL 22.
Fig. 5. C. minulissima , From the type specimens collected at Chocorua, New
Hampshire, by \V. G. Farlow, 3. Drawmgs of, a, two fructifications, xl4; h^ spores,
x510; c, a hair from outer wall of fructification, x510.
Fig. G. C. Langloisii. From the type specimens collected at St. Martinville,
Louisiana, by A. B. Langlois, cz. Drawmgs of, a, two fructifications, xl7; &, spores,
X510; r, a hair from outer wall of fructification, x510.
Fig. 7. C porrigcns. From the type specimens collected at Cinchona, Jamaica,
by W. A. and Edna L. Murrill, GOT. Drawings greatly enlarged of, a, a fructification
showing attachment to a piece of woody stem; b, diagrammatic section of the same
fructification; c, two spores, x510.
Fig. 8. C caricina. Three spores, x510, from the type specimen collected at
Verona, New York, by C. H. Peck.
Fig. 9. C cupuhiejormis. From the specimens in Ravenel, Fung. Am., 224,
collected at Darien, Georgia, by Ravenel. Drawings of, a, two fructifications, x6; 5,
a basidium, x510; c, four spores, x510.
Fig. 10. C. texensis. Three spores, x510, from the t5T)e specimens collected in
Texas, by C. Wriglit, 3779.
Fig. 11. C. funiosa. Three spores, x510, from the type specimens collected at
Aiken, South Carolina, by Ravenel, 3071.
Fig. 12. C. mcllea. From the type specimens collected at Bohemia, Louisiana,
by A. B. Langlois, 86ia. Photograph, a, of a piece of w^ood bearing many fructi-
fications, and drawings of, 6, median longitudinal section of a fructification, x60j c,
three spores, x510; d^ a hair from outer wall of fructification, x510.
Fig. 13. C. villosa. Three spores, x510, from the specimens in Krieger, Fung.
Sax., 1457, collected at Kunigstcin, Germany, by W. Ivrieger.
Fig. 14. G. Ravenelii. From the specimens in Ravenel, Fung. Am., 130, collected
at Aiken, South Carolina, by Ravenel. Drawings of, a, a fructification on a piece of
bark, x6; b, two spores, x510.
Fig. 15. C. conglobaia. From the type specimens collected at Low^er Bartlett,
New Hampshire, by R. Thaxter. Photograph, a, of a portion of a branch bearing
many clusters of fructifications, and drawings of, 6, a median vertical section through
one cluster of fructifications, x6; c, two spores, x510.
Fig. 16. C. Tilm. From specimens collected at Middlebury, Vermont, by E. A.
Burt. Photograph of, a, a piece of limb bearing many fructifications, and drawing
of, &, three spores, x510.
Fig. 17. C, fasciculata. From specimens collected at Ottawa, Canada, by J.
Macoun, 23. Photograph of, a, a piece of bark bearing many fructifications, and
drawings of, &, a cluster of fructifications, x6; c, three fructifications, xlO; d, two
spores, x510.
An?^. Mo. Rot. Gard.. Vol 1, 1914
PUME 19
1
90
b
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o o
10
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8
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9
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15
16
a
BURT — THELEPHORACEAE OF NORTH AMERICA
t. CRATERELLUS B0REALIS.-2. CYPHELLA GALEATA.-3. C. MUSCIGENA.— 4. C. CAPULA.-
5. C. MINUTISSIMA.~6. C. LANGLOISII.— 7. C. PORRIGENS.— 8. C. CARICINA.~9. C. CUPULAEFORMIS
10 C. TEXENSIS.- 11. C. FUM0SA.-12. C. MELLEA.— 13. C. VILLQSA.— 14. C. RAVEN ELIL—
15. C. C0NGL0BATA.-16. C. TILIAE.— 17. C. FASCICULATA.
COCKAYN E- BOSTON,
SOME (ENOTHERAS FROM CHESHIRE AND
LANCASHIRE '
R. R. GATES
University of London
Formerly Research Assistant to the Missouri Botanical Garden
Observations
(Enotheras are known to have been naturalized on the Lan-
cashire coast since 1805, and probably existed there much
earlier. They are now found on the sand dunes in many places,
from Liverpool and the vicinity of Birkenhead northwards
along the coast to Southport and Blackpool. They are not-
ably abundant at St. Anne's-on-Sea, where they have been
described by Bailey ('07), and in certain localities near Birken-
head (MacDouga! '07). I have grown, chiefly at the Mis-
souri Botanical Garden, extensive cultures of plants from the
latter region, from seeds obtained through Dr. D. T. MacDougal
in 1907, and have visited the Lancashire coast in 1910 and
again in July, 1914, when I travelled along the coast from Liver-
pool to Southport and from Blackpool to St. Anne's. The
Oenotheras everywhere appear to be spreading, although chil-
dren gather the flowering shoots in armfuls. The profusion of
individuals is greatest at St. Anne's, where acres of waste land
in the town are dotted over with them. Smaller colonies occur
in various other places, notably at Bidston Junction, near
Hightown and at Formby. Small groups of half a dozen plants
are sometimes found in isolated places on the dunes.
I will first refer to some of these colonies as I saw them during
my last visit, and will then describe a few of the many forms
observed in cultures.
The Bidston Junction colony, referred to in MacDougal ('07),
is a compact and almost uniform one occurring on a triangular
piece of ground between railway tracks, about five minutes*
walk down the foot path from Bidston Junction towards Wal-
lersy, on the right-hand side. Some years ago, quantities of
sand were dumped here from the coast between Wallersy and
New Brighton. Soil from neighboring gardens has also been
Issued January 30, 1915.
Ann. Mo. Bot. Gabd., Vol. 1, 1914
(383)
[Vol. 1
384 ANNALS OF THE MISSOURI BOTANICAL GARDEN
deposited here, and the advent of the Oenotheras is doubtless
from one or other of these two sources.
The plants closely resemble the ''Isle of Wight" race of CE.
Lamarckiana (to be described in a book now in process of pub-
lication) and the species as it generally appears in English
gardens. The rosettes in this colony differ in having green
midribs (both dorsally and ventrally), or pink midribs (both
dorsally and ventrally), but the depth of red varies. The same
applies to the stem-leaves. This is curiously different from
other races, such as OS. mut. rubrincrvis, in which the midribs
are red dorsally and green ventrally. The rosette leaves are
usually nearly or quite smooth, but some may be crinkled.
The plants were short, their average height being about twenty-
two inches, though some reached a height of over three feet.
L
The stems bear many red papillae. The smaller plants were
unbranched, the lower stem-leaves being closely crinkled and
curled while the upper leaves and bracts are often quite smooth,
A peculiarity of the race was the irregular disposition on the
stem of much-crinkled and nearly smooth leaves, without
gradual transitions between them such as usually occur in de
Vries's race of (E. Lamarckiana. Not infrequently crinkled
and smooth leaves alternate. The buds have fewer long hairs
than in the above mentioned race, and the sepals have uniformly
the red color pattern 5-7 of CE. mut. rubrinerviSj though they
vary somewhat in depth of shade. The dimensions of the
flowers were as follows: bud cone 50 mm., hypanthium 43 mm.,
ovary 11 mm., diameter of cone at base 11 mm., length of petals
50 mm., width 60 mm. One plant was identical with the race
of de Vries, except in its larger flowers, reddish sepals and fewer
long hairs. In most plants there is also a strong distinction
between the smooth and crinkled leaves.
This colony differs, therefore, in minor peculiarities from
any race of (E. Lamarckiana previously observed, and it ex-
hibits a relatively narrow range of variation.
Along the electric railway tracks north of Liverpool, between
Crosby and Hightown, an equally extensive and uniform colony
of CE. biennis was found. Thousands of plants, in flower and
rosettes, were growing on uncultivated land with a nearly pure
sandy soil, behind the coast range of sand hills in a long narrow
p
1914]
GATES — SOME (ENOTHERAS FROM CHESHIRE AND LANCASHIRE 385
area near a clump of small poplar trees. Near the upper end
of this area the plants differed in having smaller flowers (petals
21 mm.) and narrow leaves (20 mm. broad). The remainder of
the plants had somewhat larger flowers (petals usually 25-27
mm. long), and broader leaves (extreme width 50 mm.).^ This
was almost the only variation observed, and the race comes very
close to the type of (E. biennis L. The dimensions of the buds
were as follows: bud cone 20 mm., hypanthium 25 mm., ovary
11 mm., anthers surrounding the stigma. The rosette-leaves and
stem-leaves all have red midribs both dorsally and ventrally. On
the same stem some leaves are smooth and some more or less
crinkled. The buds are green, devoid of red, with some long
hairs, and there are no red papillae on any part of the plant.
Some of the larger plants are well-branched and with very stout
stems, a huge pith and a very narrow ring of wood.
This colony is even more uniform than the previous one,
and must have originated from one or a very few plants.
Small colonies of CE. biennis were seen at Formby, near the
station and in other places. A race of (E. Lamar cUana also
grows here on the dunes, although I did not succeed in finding
the spot, but local gardens cultivate it. The species is depicted,
however, in a rose window erected in St. Luke's Church, Formby,
in 1898, containing representative plants of the local flora.
The central portion of the window is divided hexagonally and in
the six sections the evening primrose alternates with the sea
holly. The foliage and large flowers of the former are distinctly
shown. Around the margin of the window are Pyrola rotundi-
folia and irises.
At Blundell Sands, near Crosby, a small colony of (E. Lamarck-
iana was seen on waste ground, and again on the extensive
sand dunes between Birkdale and Ainsdale, near Southport.
In the latter case there were only three plants, and these pos-
sessed red sepals, color pattern 7, green midribs, crinkled leaves,
and about - long hairs.
4
By far the greatest abundance of plants was found at St.
1 These apparently correspond to Lysimachia virginiana altera, Joliis latiorihus,
floribus luteis majoribus, Cat. AltdorfF. See Gates, R. R. The mutation factor
in evolution [pp. 61, 65, 70]. Macmillan. London.
[Vol. 1
386 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Anne's. In addition to those in the town, which are in great
profusion, numerous smaller colonies are scattered along the
adjacent sand dunes. The great majority of the plants is the
same as at Bidston Junction except in the crinkHng of the leaves,
having foliage closely resembling that of de Vries's (E. Lamarck-
iana, midribs red both above and below, the red absent in
some individuals. The flower measurements were, length of
petals 50 mm., hypanthium 45 mm., ovary 10 mm. Several
aberrant individuals were also observed. One dwarf mutant
was found growing in the shade of a large plant. It resembled
(E. mut. nanella but had red midribs. One large rosette, having
leaves very obtuse and pale pink midribs, probably belonged to
(E. mut. hrcvistylis. A number of plants represented a shorter
spindling type with very narrow rosette-leaves (18 mm. wide
X 14 cm. long). Another plant belonged to a new type, large
and branching with thicker, narrower leaves (33 mm. x 13 cm.),
stiffer and narrowly pointed, midribs white, and later in beginning
to flower (buds only half developed, July 16).
In addition to these probable mutants, there were found in
one field a few plants of a small-flowered (E. biennis race grow-
ing with the (Z;. Lamarckiana. They differed from the latter
only in the small flowers (petals 22 mm., style short), and hence
were unlike the (E. biennis race previously described. Near by
were also found plants, evidently hybrids of these two races,
with petals about 30 mm. in length.
Cultures
Some of my cultures of Oenotheras from near Birkenhead have
already been described in a general way (Gates, '13). Here I
wish to describe a few of these forms in detail, and also to refer
to my experiments with plants from St. Anne's. I have not
seen the colony from which the Birkenhead seeds were obtained,
but it evidently contains a great profusion of forms belonging
to both (E. Lamarckiana and (E. grandiflora, while all the
colonies I have observed have a much more uniform Donulation
CE. MULTIFLORA
One of the distinct races in these cultures I have already
(Gates, '10) referred to as (E, multiflora. It is descended
1914]
GATES — SOME (ENOTHERAS FROM CHESHIRE AND LANCASHIRE 387
■
entirely from one individual from a sowing of Birkenhead seeds
at Woods Hole in 1908. From this individual an Fi of 376
plants was grown in the two following years. About 4 per cent
of these plants showed viresccnce, as described in the above
paper. In 1910 a total of 297 plants were grown, most of which
belonged to the Fa. An F3 numbering 193 plants in nine fam-
ilies was grown in 1911, and an F4 of 356 plants in eight fam-
ilies in 1912. The plants were by no means uniform, and they
varied considerably from year to year. The description given
is therefore a generahzed one, and the condition of variability
is no doubt similar to that of many wild ''species." By isol-
ating the offspring of a larger number of individuals, no doubt
this variation could have been further analyzed, but more
pressing problems have prevented this being done.
Plate 20 fig. 1 shows a typical rosette of my 1909 culture,
pi. 20 fig. 3 the full-grown plant, and pi. 20 fig. 6 a flowering
shoot on a larger scale. Specimens of this species are preserved
in the herbarium of the Missouri Botanical Garden from my
cultures of 1909, and in the British Museum (Natural History)
from the 1912 famiUes.
Description : Rosette of few leaves, broad and obtuse-pointed,
somewhat crinkled. Full-grown plant pyramidal in outline,
with lateral branches and persisting rosette leaves. Average
height about 88 cm. Stems slender, stem-leaves smooth, lance-
olate, bracts broadly cuneate at base with a very short petiole,
tip long-pointed, more or less curled, margin irregularly repand-
denticulate. Inflorescence compact, flowers numerous; buds
squarish, slender with very long and slender sepal tips, sepals
thin, bud cone 35 mm. long, hypanthium 37 mm., sepal tips 7
mm.', ovary 10 mm., petals 43 mm., very broad and overlapping
when flower is open, long hairs fairly numerous. Few red
papillae on main stem, many on side branches. In 1909 culture
the buds were all green, but in 1911 they had the red color
pattern of (E. mut. ruhrinervis and the stems were also reddish.
As regards variations, virescence appeared in the first two
generations but not in the last two. On the other hand, a var.
elUptica was first observed in F2 and further studied in F3 and
F4. This variety differs essentially in being smaller and having
Tiflrrnwpr leaves and narrow, more or less elUptical petals. Plate
[Vol*. 1
388 ANNALS OF THE MISSOUBI BOTANICAL GARDEN
20 fig. 2 shows a rosette of this variety in Fa (1911). One family
of 50 plants in 1910 contained 5 of this variety. Usually these
plants show partial variability, some flowers having broad
petals and others narrow and elliptical ones. Even the differ-
ent petals of the same flower may show these differences.
Flowers with elliptical petals are invariably smaller and are fre-
quently found on the side branches when those of the central
stem have normal petals. Hence this variation may be a matter
of strength in the plant. The variation, from petals which are
broad and truncate or emarginate to those which are narrow
and elliptical, or even almost cruciate, is continuous. Thus
on one plant in 1911, the dimensions of the petals in two flowers
were as follows:
Flower 1. Petal (1) 31 mm. x 21 mm.
Petal (2) 25 mm. X 17 mm.
Petal (3) 20 mm. X 12 mm.
mm
In this flower the petals are very small and
size but all elliptical.
Flower 2. Petal (1) 38 mm. x 39 mm.
Petal (2) 37 mm. x 37 mm.
Petal (3) 34 mm. x 36 mm.
Petal (4) 35 mm. x 36 mm.
In this flower the petals were nearly full size, :
scarcely elliptical.
this condition is on a sliding
with only broad petals giving some offspring with elliptical
petals, and plants with elliptical petals giving some offspring
having only broad petals, though in the latter case the plants
bearing elliptical petals are more numerous than in the former
case. Thus the F3 family from a normal plant contained 14
specimens having broad petals only and 15 having some ellipt-
ical petals ; while another F3 family of 44 plants derived from a
plant having elliptical petals contained only 5 plants having
exclusively broad petals. These peculiarities of the petals are
probably to a large extent under the control of environmental
features such as temperature and water supply.
The difference between broad and narrow leaves is much
sharper. Thus in my F4 cultures in 1912 certain families contain
1914]
GATES — SOME CENOTHERAS FROM CHESHIRE AND LANCASHIRE 389
both the broad or normal type (pi. 20 fig. 5) and the elliptica
variety (pi. 20 fig. 4). The latter had a number of flowers with
elliptical petals and it also had a different method of branching.
Plate 21 fig. 12 is representative of a uniform F4 culture of 49
plants of the variety elliptica. This photograph is taken on a
larger scale, and the nodding of the stem is merely due to wilting.
This differs from typica (pi. 20 fig. 5) constantly in having nar-
rower leaves and short branches, as well as in the occasional
elliptical flowers which appear to be largely under environ-
mental control.
The variability of this race is therefore as interesting as are
the features, such as the general bud and leaf characters, in
which it is constant. The fact should also be mentioned that a
lata-Wke mutant, doubtless having 15 chromosomes, appeared in
the F4 generation, and also a mutant resembling CE. mut. albida.
CE. EUBKINEEVOIDES
This race resembles CE. mut. ruhrinervls in many features,
and yet differs from it constantly throughout. I have pre-
viously referred to this Birkenhead race as No. 25 (Gates, '11,
p. 350) and studied the variation of the red stripes on the buds.
In all, 1968 plants of this race have been grown in the years
1909-1912, so that four generations of offspring from a single
individual have been cultivated. An illustration of that indi-
vidual has already been published (Gates, '12, pi. 3). One fam-
ily of offspring was grown in 1909, two in 1910, eight in 1911 and
nine in 1912. Usually the variability of families progressively
decreased, since each family was derived from the selfing of
previous veneration. The discussion of
individual of the
the precise ancestry of this race is of course out of the question,
but Its characters bear nearly though not quite the same rela-
tion to the (27. Lamarckiana from this region that the Lamarck-
iana and ruhrinervis of de Vries's cultures bear to each other.
The 1909 family, or Fi, numbered 111 plants. Plate 21 fig. 8
shows one of these as a rosette. The leaves are narrower and
more pointed than in mut. ruhrinervis, and nearly smooth.
About 20 of the plants in this culture omitted the rosette stage
altogether and shot up a stem directly from the seedhng stage
(pi. 20 fig. 7). A normal mature plant of this family is shown
8
[Vol. 1
390 ANNALS OF THE MISSOURI BOTANICAL GARDEN
in pi. 21 fig. 11. It will be seen that there is no indication of a
rosette, and the branching is quite different from that of (E.
nmt. ruhrinerms. In many cases, however, a rosette is formed.
When the rosette is omitted the branching is changed. Plate
21 fig. 10 shows on a larger scale another individual in flower.
The stem-leaves differ from those of (E". mut. ruhrinerms in being
narrower, more pointed and smoother.
In this race the red papillsc on the stem were very numerous,
and the buds Hkewise were slightly more red than in (E. mut.
rubrincrvis. The modal color pattern of the whole population
was 5 as in 0^. mut. rubrincrvis, but plants with their mode at
7 were much more numerous than in the latter (see Gates, '11,
p. 351). The race as a whole inherited the capacity for pro-
ducing a slightly greater amount of pigment. The ovary usu-
ally bore many long hairs arising from red papilla); on the hy-
panthium were few long hau's from slight green mounds; and
on the bud cone scattered long hairs from conspicuous red
papilla). In occasional buds, when the color pattern was only
3, the green papilla) were more numerous. In addition to the
color pattern of the sepals there was usually weak red on the
1
hypanthium.
The same conditions as regards pigmentation have been main-
tained in later generations. The plants WTre, however, by no
means uniform in all respects, and this was not to be expected
since they were derived from one individual of a freely inter-
crossing population. Plate 21 fig. 9 represents a rosette of
one of the F2 plants. The latter differs obviously from the one
represented in pi. 21 fig. 8, but the race retained in this and
subsequent generations the long, narrow, smoothish leaves as
well as the pigmentation. The various F3 and r4 famihes, each
derived from a selfed individual, produced sub-races differing
more of less from each other and varying within narrower limits.
It docs not appear that the Mendelian theory of the sorting out
of factors, or ''genes," affords an adequate explanation of all
these phenomena.
' Since this condition of bud-pignieiitation resembles that obtained in certain F|
and F3 hybrids of (E. mut. ruhricalyx and (E. grandiflora (see Gates '14), it ie possi-
ble that it may have arisen in a similar way, i. e., by the appearance of a red-budded
mutation which subsequently crossed with other species, in which crosses some blend-
ing of pigmentation occurred giving rise to the present condition.
1914]
GATES — SOME ODNOTHERAS FROM CHESIIIIiE AND LANCASHIRE 391
CE. TARDIFLORA
This name I have used for another race having many pecuUar-
ities and showing more resemblance to (E. grandiflora in its
m
flowers and foliage. It is race No. 52 from the same source as
the above. A single individual produced in 1909 nineteen plants
which were fairly uniform. The rosettes contained only a few
leaves, but large plants w^ere formed, one of which is shown in .
pi. 22 fig. 17. Although this photograph w^as taken on August
21, the plants with one exception had not begun to flower. The
leaves resembled those of (E, grandiflora. They were large
with long and acute tips, tapering to the bases, often bearing
reddish blotches, sometimes much curled, somewhat crinkled
along the midrib. The margin was conspicuously serrately
toothed (see pi. 22 fig. 17). At the end of the season (Septem-
ber) these plants came into bloom, and pi. 22 fig. 20 shows a
plant photographed on October 2. The buds resembled those
of (E. grandiflora but were small. The bud cones were pointed,
smooth and rounded, the petals slightly larger than in (E. bien-
nis, or in a few cases much larger. The petals were also deeply
emarginate, strongly cuneate and narrow; and the bracts were
very small, narrowly lanceolate and yellowish, giving a peculiar
appearance to the flowering shoot. The margins of the bracts
were nearly entire or in some cases distantly denticulate.
The offspring of the plant in pi. 22 fig. 20 were grown and
showed the same peculiarities. The race has not been culti-
vated further. It was doubtless of hybrid origin and was more
nearly allied to (E. grandiflora than to the Lamarckiana complex.
CE. RUBRITINCTA
Reference may be made to one further race which was known
as ''type m." It originated from one plant in a sowing of the
Birkenhead seeds in 1909. It will be understood that scarcely
two plants from this sowing were alike, but some were much
more distinct than others. The plant in question was a hand-
some one with very narrow leaves and bright red midribs. Its
offspring, grown in 1911, were lost with the exception of one
-.
plant which was the same as the parent. It is shown in pi. 22
fig. 16. The basal leaves were very long with long petioles, the
stem leaves very narrow, smooth, with margin closely repand-
[Vol. 1
392 ANNALS OF THE MISSOURI BOTANICAL GARDEN
denticulate, blade narrowing gradually to a very short petiole,
midribs and petioles bright red dorsally and ventrally; lower-
most bracts 17 mm. in width by 9 cm. in length, upper bracts
11 mm. wide by 58 mm. in length. "^The buds most resemble
those of CE. grandijlora, being nearly devoid of long hairs, slender
and somewhat rounded, with setaceous sepal tips and some red
on the sepals; length of petals 32 mm., hypanthium 43 mm.,
sepal tips 9 mm., ovary 10 mm.
In 1912 three families of Fa offspring, numbering in all 236
plants, were grown from the plant just described. All three
families agreed in containing several types exhibiting a remark-
able degree of variability.
An attempt was made to place the plants in five classes, but
the categories overlapped and made classification for the most
part impossible. The majority of the plants resembled the
parent individual in their main features but they varied enor-
mously in width of leaf from broad (21 mm.) to very narrow
-6.5 mm.). These conditions were connected by interme-
diates, and, moreover, there were considerable variations within
the individual, one branch with very narrow leaves being found
on a plant with broad leaves. In addition to these variants,
the three famiUcs contained 35 dwarfs, or 14.8 per cent, and
the latter varied in leaf- width in the same remarkable manner.
The dwarfs agreed only in having short internodes. Two of
them are shown in pi. 21 figs. 13, 14, the former having narrow
leaves and extremely short internodes, the leaves of the latter
being quite linear. The plant would never be taken for an
cenothera.
The advent of a large percentage of dwarfs in this family is
similar to their occurrence in other (E. grandijlora races from
that locality (see Gates, '14, p. 246). The precise manner in
which this capacity for producing dwarfs is inherited, is a diffi-
cult question which need not be considered here, particularly
as it has been discussed elsewhere (Gates, '14).
Plate 22 fig. 15 represents one of the LamarcJdana-\ike
rosettes from this source, grown in 1909. Others approached
de Vries's race more closely, to the point of identity. Plate 22
figs. IS, 19 represent selected rosette-leaves taken from this cul-
ture to show the range of types exhibited. Such leaves as the
GATES— SOME (ENOTHERAS FROM CHESHIRE AND LANCASHIRE 6J6
two on the right in ph 22 fig. 18 were greatly overgrown and
were far larger than ever appear even in (E. mut. gigas. These
forms have not been sufficiently studied since to give an ade-
quate account of them.
It will be obvious that the forms described here under the
names multiflora, multijiora elliptica, ruhrinervoides , tardiflora
and rubritincta are not pure species or even true-breeding races.
They are undoubtedly as diverse from each other as average
species, however, and many systematic species if bred experi-
mentally would probably not breed true within narrower hmits
than these races have done. One feature of interest attaching
to these races is the fact that the main type persists essentially
unchanged, though various mutants and heterozygous forms
are thrown off. The behavior is not, in the main, hke the Men-
delian process of recombination. Repeated selfing of each race
usually decreases its variability by eliminating various hybrid
elements. But this process does not extend to the basal differ-
ences between the races, which, as we have seen, remain as
unhke as they were before. In this aspect the hereditary
behavior of these races resembles that of CE. Lamarckiana.
But there are a number of differences which I need not fully
consider. Thus (E. muliifl
elliptica
much as though it were split off from a heterozygous condi-
tion, and the variabihty of rubritincta in leaf-width, as well as
its production of numerous dwarfs, is unlike anything in the
behavior of (E. Lamarckiana.
Many other equally distinct types were derived from this
locality (see, e. g., pi. 22 figs. 18, 19), but they have not been
cultivated in subsequent generations.
CE. LAMARCKIANA FROM ST. ANNe's
In 1910 I obtained seeds from a colony of (E. Lamarckiana
growing by the Manchester Children's Hospital Convalescent
Home, at St. Anne's-on-Sea. Many of these were found in
later cultures to agree exactly with the Lamarckiana of de Vries
except in the red color pattern of the sepals. I was formerly in-
clined to lay little stress on this difference but there is no doubt
that it is inherited. The fact therefore remains that a precise
duplicate for de Vries's race of (E. Lamarckiana is relatively
[Vol. 1
394 ANNALS OF THE MISSOURI BOTANICAL GARDEN
infrequent on the Lancashire coast, although many forms ap-
proach it very closely and differ only in this one feature. As
will be seen below, certain other plants agreed with de Vries's
Lamarckiana except in the shape of the buds.
In 1911 a sowing of the seeds yielded 22 plants. The rosettes
were for the most part uniform and very similar to (E. La-
marckiana, two, however, having red midribs and lighter green
leaves {rubrinervis type). One plant was aberrant, resembling
(E. mut. semilata in its buds, which were, however, small as in
(E. biennis. The bud cone was also somewhat rounded and
barrel-shaped, length of ovary 11 mm., hypanthium 37 mm.,
cone 19
short so that anther
round base of stigma. The features of this plant make it
scarcely likely that it arose as a hybrid. It produced plenty of
pollen and seeds.
Another sowing of these seeds in 1912 yielded 140 plants,
which included one mut. lata with bad pollen (doubtless having
15 chromosomes) and one variegated Lamarckiana plant. The
variegation was noticed when the plant was a young seedling.
It reached maturity and proved to be a periclinal chimera.
Nearly all the leaves were variegated green and yellow. Many
leaves were green bordered with yellow, showing the absence of
chloroplasts from the epidermal and probably also the hypo-
dermal layer. Occasional leaves were almost entirely yellow,
and some were yellow on one side of the midrib and green on
the other. There were also broad white bands on the
of the sepals. The pollen was abundant and plentv of seed
mar
were set.
Two sowings of seeds from this plant were made in 1912.
The seeds numbered respectively 121 and 145. Only two seeds
in one pan were observed to germinate, and the seedlings quickly
died, probably from lack of chlorophyll. Regarding the origin
of this periclinal mutation, it would appear to have originated
fr
mbryo after
The foliage in the rest of the culture agreed with the type of
(E. Lamarckiana. One plant differed in having stem-leaves more
or less pointed at the base, not crinkled, midribs pink, and smaller
flowers (petals 29 mm. long x 38 mm. broad, style short, buds
1914]
GATES — SOME CENOTHERAS FROM CHESHIRE AND LANCASHIRE 395
squarish) . Two other plants agreed exactly with CE. Lamarck-
iana except in the buds. The petals were 35 mm. long x 48
mm. broad, emarginate, anthers reaching nearly to top of stigma
lobes, sepals green and with the same pubescence as in (E.
Lamarckiana, from which these two plants therefore differed
only in the somewhat smaller flowers and shorter style. One
mut. nanella also occurred in this culture, and several other
slightly aberrant individuals, including a plant with broadly
elliptical foliage. The '^ Lamarckiana foliage" was also more
variable than in cultures from de Vries, this no doubt being due
to the continued inbreeding in the latter case.
It will be understood that the new forms described here are
scarcely to be looked upon as ''new species" according to the
usual interpretation at the present time. They merely represent
a partial analysis of a complex interbreeding colony of forms,
and their variability is one of their most interesting features.
Nearly all if not all the differences observed are inherited, how-
ever, and the mutations can in many instances be separated
from the characters arising through hybridization. The forms
are. moreover, as distinct from each other as many species of
(Enothera.
In conclusion, I am indebted to the Missouri Botanical Garden
and the John Innes Horticultural Institution for the facilities
provided for
and to Mr. E. J. Allard for
of the photographs. A portion of the expenses of my
second visit to Lancashire was defrayed by a grant from the
Royal Society.
LiTEEATURE CiTED
Bailey, C. ('07). De Lamarck's evening primrose {(Enothera Lamarckiana) on the
sandhills of St. Anne's-on-the-Sca, North Lancashire. (Address, Annual Meeting
Manchester Field Club.) 1-28. pi. 1-6. 1907.
Gates, R. R. ('10). Abnormahties in (Enothera. Kept. Mo. Bot. Gard. 21: 175-84.
vl. 29-31 . 1910.
ffino
thera. Zeitschr. f. induct. Abst.- u. Vererbungsl. 4: 337-72. pi. 6. J. 1-5.
1911.
• ('12). An onagraceous stem without internodes. New Phytologist 11:
50-53. vl. 2-3. 1912
, ('13). A contribution to a knowledge of the mutating Oenotheras. Trans
Linn. Soc. Bot. 8: 1-67. pi. 1-6. 1913.
O\)0 ANNALS OF THE MISSOURI BOTANICAL GARDEN
xperiments
are independent phenomena. Zeitschr. f. induct. Abst- u. Vererbungsl. ii:
/. 1~S5. 1914.
MacDougal, D. T., Vail, A. M., and Shull, G. H, ('07). Mutations, variations, and
relationships of the Oenotheras. Carnegie Inst, of Washington PubL 81 : 1-92
pi 1-22. /. 1-7S. 1907.
Explanation of Plate
PLATE 20
Fig. 1. (E. mnUiJlora, rosette, 1909
Fig. 2. (E. ynuliijlora elliptica, roset
Fig. 3. (E, multijlora, full-grown pi
Fig. 4. (E, multijlora elUplica, 1912
Fig. 5. (E, multijlora, 1912.
Fig. 6. (E. multijlora, flowering she
Fig. 7. (E. rubrinervoides, vounc: n]
Ann. Mo. Bot. Gard., Vol 1. 1914
Plate 20
vA,
3
\
6
GATES— OENOTHERAS
COCKAYNE, BOSTON,
(Vol. 1, 1911
398 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 21
Fig. 8. (E. rubrinervoidcSy rosette, 1909.
Fig. 9. ffi. ruhrincrvoidesj rosette, 1910.
Fig. 10. (E. ruhririervoideSj Bhowiiig nearly smooth, pointed leaves, 1909.
Fig. 11. (E. ruhrinervoideSj no rosette, 1909.
Fig. 12. (E, mtfJtijlora ellipticaj 1912. (Tip of plant drooped from wilting.)
Fig. 13. Linear-leaved dwarf in offspring of CE. rubritincta, 1912.
Fig. 14. Dwarf offspring of (E. rubrilinda, 1912.
Ann. Mo. Bot. Gard., Vol 1, 1914
Pl,\te 21
10
n
14
GATES— OENOTHERAS
L> A "^.J
400 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1, 1914J
Explanation of Plate
PLATE 22
Fig. 15. (E. Lamarckiana-liko rosette, 1909.
Fig. 16, CE. rubritincta, 1911.
Fig. 17. ffi. tardiflora, showing serrated leaves and absence of flowers, August
21, 1909.
Fig. 18. Selected leaves from various rosettes, 1909.
Fig. 19. Selected leaves from various rosettes, 1909.
Fig. 20. (E. tardiJIorUj showing late appearance of buds, October 2, 1909.
Ann. Mo. Bot. Card., Vol t, 1914
Plate 22
GATES— OENOTHERAS
C0CKAY>4E:, 30ST0N,
A TEXAN SPECIES OF MEGAPTERIUMi
R. R. GATES
r
University of London
Formerly Research Assistant to the Missouri Botanical Garden
While looking over some material in the herbarium of the
Missouri Botanical Garden, a sheet was found containing three
specimens which were so distinctive that it seemed desirable
to describe them. The interest in them was enhanced by the
fact that one of the specimens differs strikingly from the other
two in such a wav as to suggest that it may be a mutation.
The
collected at Amarillo Creek
Northern Texas, by J. Reverchon, who had recognized them a:
representing a new species of Megapterium.
I am indebted to Dr. Greenman for suggesting a very appro
priate name for this species. The diagnosis is as follows:
Megapterium argyrophyllum, sp. nov. Plate 23. figs. 1 and 2
Herba csespitosa; foliis lanceolatis, petiolatis obscure gland
uloso-denticulatis, utrinque dense canescento-pubescentibus
caulibus et alabastris (hypanthio et ovario incluso) canescenti
pubescentibus; ovarium quadrialatum, pedicellatum; hypan
thium 9-10 cm. longum, paulatim ad basin coni dilatum
petala 3-4 cm. longa.
Var. retusifolium
Plate 23. fig
A forma typica differt foliis subrotundis bis oblongo-obo-
vatis, retusis, mucronatis; flora? grandiorai (petala 45 mm.
longa).
Specimens examined:
Texas: on rocky bluffs at Amarillo Creek, in northern Texas,
29 May, 1902, /. Reverchon, 2749 (Mo. Bot. Gard. Herb.),
type; stony bluffs along Red River, Randall Co., northern
Texas, 12 August, 1900, H. Eggert (Mo. Bot. Gard. Herb.,
4 sheets).
Two of the specimens, one slightly older than the other (see
pi. 23 fig. 1, 2), represent the type of the species. The plants
are cffispitose or with very short internodes, leaves coriaceous,
lanceolate, broad-pointed, tapering below to a petiole, about
» Issued January 30, 1915.
Ann. Mo. Bot. Gaed., Vol. 1, 1914
(401)
[Vol. 1
402 ANNALS OF THE MISSOURI BOTANICAL GARDEN
8 cm. long by 2 cm. in greatest width, margin distantly and
obscurely glandular -denticulate, very densely and uniformly
covered on both surfaces with an appressed canescent pubescence
of long, pointed, tuberculate hairs. Stems and buds less densely
covered with the same type of pubescence, ovary four-winged,
10-15 mm. in length, densely canescently pubescent, pedicel-
late; hypanthium 9-10 cm. in length, 2-2.5 mm. thick, grad-
ually widening to base of cone; bud cone 30-35 mm. in length,
diameter at base 8 mm., sepal tips appressed, 3-4 mm. in length,
petals 3-5 cm. long, stigma surrounded by or slightly exceeding
the stamens; capsules immature.
The remarkable canescent pubescence covering the whole
plant, as well as the caspitose habit, distinguish this species
from Mcgapterium missouriensis (Sims) Spach, and M. macro-
carpum.^ The flowers are also smaller, there are no purple
spots on the sepals, and the hypanthium is shorter than in
these species, which differ in foliage as well. The present
species is apparently perennial. Its nearest relative is M.
Frernontii (Watson) Britton, from which it differs in the more
ca^spitose habit, larger flowers, and much broader leaves.
The variety reiusifolium is founded on the third specimen on
the sheet (see pi. 23 fig. 3). It differs sharply from the species
in the shape of the leaves, which arc very broad and blunt at
the point, subrotund to oblong-obovate, retuse, and distinctly
mucronate. The margin of the leaves is also nearly or quite
entire. The flowers are larger (petals 45 mm., bud cone 9 mm.
in diameter at base). Microscopic examination of the hairs
disclosed considerable variation in size, but apparently no con-
stant difference from those of the species.
The Eggert specimens, while obviously belonging to the same
species, show much more variability in fohage. The leaves
specimens from
mm
in width) to broad oblong- lanceolate (30-36 mm. mde) and
acuminate. The latter resemble var. retusifolium except the
leaf tips, which are only sHghtly retuse in one specimen. One
of the broad-leaved specimens also has a smaller flower (petals
20 mm.). Cultures from seeds from this locahty would doubt-
^ Megapterium macrocarpum (PursJi), comb. nov.
Oenothera macrocarva PnrsTi. Fl Am Spr^f -»• T'ii i«i/i
1914]
GATES — A TEXAN SPECIES OF MEGAPTERIUM 403
less disclose a considerable number of forms. The ripe fruits
from these specimens are broadly winged, nearly orbicular,
about 35 mm. long and 25 mm. wide, retuse or acuminate at the
apex.
Examination of herbarium specimens of M. missounensis
(Sims) Spach makes it evident that the polymorphism in this
species as now understood is quite as great as in many species
of CEnothem. There are included races varying in amount and
character of pubescence, in width of leaf from broadly lanceolate
to almost linear, in presence or absence of purple spots on the
senals. in size of flower, and other features.
[Vol. 1. 19HI
404 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 23
P'igs. 1 and 2. Megaplerium argyrophyllurn. From the type specimens, J. Rever-
chon, No. 2749 in part, in the Herbarium of the Missouri Botanical Garden.
Fig. 3. M. argyrophylluin var. retusifolium. From the type specimen, J. Rever-
chon No. 2749 in part, in the Herbarium of the Missouri Botanical Garden.
Anm. Mo. Bot. Garp.. Vol 1. 1914
Plate 23
GATES—MEGAPTHRrJM
COCKAYNE., BOSTON-
DIAGNOSES OF FLOWERING PLANTS, CHIEFLY
FROM THE SOUTHWESTERN UNITED STATES
AND MEXICO 1
i
J. M. GREENMAN
Curator of the Herbarium of the Missouri Botanical Garden
Associate Professor in the Henry Shaw School of Botany of
Washington University
AND C. H. TPIOMPSON
Assistant Botanist to the Missouri Botanical Garden
The present paper is the result of a study of several collec-
tions of plants from the southwestern United States and Mexico,
especially the relatively large series of specimens secured by Mr.
Harley P. Chandler at Rio Hondo, Texas, and by Mr. Charles
Russell Orcutt along the Texas-Mexican boundary and in vari-
ous parts of Mexico. These collections have been received at
the Missouri Botanical Garden for identification and incidental
to the work thereto the following plants seem to the writers to
be worthy of record and characterization.
Anthericum (Hesperanthes) Chandleri Green man & Thomp-
son, sp. nov.
Fibrae radicales carnosse apice clavatse, collo radicis parce
fibroso; foliis plurimis 12-15 graminoidcis planis lanceolato-
linearibus sensim attenuatis acutis 3.5-4.5 dm. longis 7-10 mm.
latis circiter 24-nerviis cum venis transversis conjunctis utrinque
glabris integerrimis; scapo 1 m. alto tereti glabro bracteato,
bracteis plus minusve foliiformibus sursura gradatim reductis;
inflorcscentiis paniculatis usque ad 3.5 dm. longis glabris, racemo
terminah 2-2.5 dm. longo, racemis lateralibus 1-1.5 dm. longis,
bracteis triangulari-acuminatis vel lanceolato-attenuatis sub-
scariosis 3-20 mm. longis; floribus 2-4 in axillis bractearum;
pedicellis 10-12 mm. longis infra medium articulatis; perianthio
palhdo-flavo vel stramineo, laciniis oblongo-lanceolatis triner-
viis circiter 1 cm. longis; staminibus perianthio duplo breviori-
bus, filamentis muricatis; stylo 8 mm. longo glabro; capsula
matura ignota.
Issued January 30, 1915.
Ann. Mo. Bot. Gaed., Vol. 1, 1914
4
(405)
406 ANNALS OF THE MISSOUEI BOTANICAL GARDEN
[Vol. 1
Specimen examined:
Texas: vicinity of Rio Hondo, Cameron County, September,
1913, Barley P. Chandler, 7059 (Mo. Bot. Card. Herb.),
TYPE.
This species belongs to the subgenus Hesperanthes according
to Baker's treatment of this group (Jour. Linn. Soc. Bot. 15:
253-363.1876); it is apparently most nearly related to A.
stenocarpum Baker, a co-type of which is in the herbarium of
the Missouri Botanical Garden, from which it is readily distin-
guished by the broader leaves, entire leaf-margins, the presence
of anastomosing cross-veins, and by the leafy scape and yellow
flowers.
Zephyranthes chrysantha Greenman & Thompson, sp. nov.
Bulbus subglobosus 2-2.5 cm. diametro tunicis brunneo-
nigrescentibus vestitus, collo 3-5 cm. longo 6-8 mm. diametro;
fohis 2-4 sub anthcsi evolutis linearibus 2.5-4.5 dm. longis 2-3
mm. latis glabris; scapis 2-3 dm. altis glabris; spatha mem-
branacea 2.5-3.5 cm. longa inferne tubulosa, tubo 1-1.5 cm.
longo, lobo unilaterah lanceolato 1.5-2 cm. longo; pedicelHs
2.5-3.5 cm. longis gracihbus; perianthio infundibuhformi 3-3.5
cm. longo flavo 6-lobato, tubo cylindraceo circiter 5 mm. longo,
lobis oblanceolatis 3-3.2 cm. longis 5-12 mm. latis acutis
staminibus ad apices tubi perianthii insertis segmentis perian-
thii duplo brevioribus; stylo brevitrilobato staminibus suba;-
quantibus; capsula depresso-globosa 10-12 mm. longitudine et
diametro, seminibus numerosis irregulariter compressis 5-6 mm.
longis 2-5 mm. latis atratis et sa^pe nitidis.
Specimen examined:
Texas: Rio Hondo, Cameron County, September, 1913, Harley
P. Chandler, 7056 (Mo. Bot. Card. Herb.), type.
The species here characterized is allied to Z. Eggersiana Urb.,
particularly in the size and color of the flowers, but differs in
having more numerous and broader leaves, shorter perianth-
tube and longer spathes.
Sisyrinchium angustissimum (Rob. & Greenm.) Greenman &
Thompson, comb. nov. Plate 24.
S. alatum Hook, var.? angustissimum Rob. & Greenm. Am.
Jour. Sci. 50: 166. 1895.
Radices carnoso-fibrosi fasciculati : cauHbus erectis strictis vel
\
i
i
*
u
T
ji t
1014]
GREENMAN AND THOMPSON — FLOWERING PLANTS 407
subflexuGsis 2.5-9 dm. altis multo-ramosis angustissime ancipiti-
alatis foliosis glabris vel obscure hirtello-puberulentis basi
reliquiis brunneis fibrosis squamarum et foliorum primorum
obtecto; foliis radicalibus linearibus gramineis usque ad 4.5 dm.
longis 1-4 (rarius 6) mm. latis crebrenerviis glabris vel margin-
ibus hirtellis, eis caulinis conformibus sed sursum gradatim
reductis; spatha diphylla, bracteis foliiformibus 1.5-2 cm. longis
glabris marginibus plus minusve purpurascentibus, pedicellis
2-4 ex eadem spatha 1.5-2.7 cm. longis gracilibus glabris; perian-
thio profunde 6-partito verisimiliter flavo, lobis ovato-ellipticis
acutis vel emarginatis et submucronatis 5-7-nerviis; ovario
oblongo-obovato juventate ssepe pubescenti glabrato; capsula
matura oblonga 5-10 mm. longa 4-6 mm. diametro glabra,
seminibus subglobosis circiter 1.5 mm. diametro in sicco nigres-
centi.
Specimens examined:
Mexico: State of Oaxaca, Sierra de San Felipe, altitude 2895
m., 22 June and 29 August, 1894, C. G. Pringle, 4703 (Mo.
Bot. Gard. Herb.), co-type; Sierra de San Felipe, altitude
3048 m., August-September, 1894, Charles L. Smith, 758
(Mo. Bot. Gard. Herb.). State of MoreloS; lava beds
above Cuernavaca, altitude 2590 m., 19 November, 1902,
C. G. Pringle, 11191 (Mo. Bot. Gard. Herb.). State of
Puebla, vicinity of San Luis Tultitlanapa, near Oaxaca,
June, 1908, C. A. Purpus, 3356, 3357 (Mo. Bot. Gard.
Herb.).
After a careful reexamination of the original material on
which this variety was based, particularly in the light of addi-
tional specimens from subsequent collections, it seems unde-
sirable to retain the plant as a variety of S. alatum Hook. Mr.
Hooker's species was founded on specimens collected in Deme-
rara, British Guiana, by Dr. Hancock; and specimens secured
by Mr. Gardner in the Organ Mountains of Brazil and by
Tweedie on the marshes of the La Plata River were considered
conspecific. While the writers have not seen any of these
specimens, yet from the original description and the illustration
accompanying it that species is interpreted as having a broadly
winged stem, short and relatively broad ensiform leaves, and
broad spathes. These characters can not be applied properly
[Vol. 1
408 ANNALS OF THE MISSOURI BOTANICAL GARDEN
to the Mexican plant in question. It seems advisable, there-
fore; to regard the south Mexican plant as a distinct species
which mav be further characterized as above.
OECOPETALUM Greenman & Thompson, gen. nov. Icacinaccce
Calyx 5-lobus. Petala 5 hypogyna valvata intus costata,
margine et apice inflexa. Stamina 5 hypogyna pctalis alterna
et iis basi cohscrentia; filamentis dilatis glabris apice contractis;
anthera? erectse lanceolatae basi sagittatse connectivo latiusculo;
thecse lateralse remota^ et in cavitatibus petalorum receptee.
Discus obsoletus. Ovarium uniloculare, stylus erectus conicus,
stigma tcrminale. Ovulum 1 pendulum. Fructus ignotus. —
Frutices vel arbores. Folia alterna coriacea integerrima.
Florcs cymis brevibus axillaribus dispositi.
O. mexicanum Greenman & Thompson, sp. nov. Plate 25.
Frutex (?) vel arbor (?); ramis cortice griseo tectis; ramulis
juvcntate sericeo-pubescentibus mox glabratis; foliis alternis
petiolatis eHiptico-lanceolatis 1-2.5 dm. longis 3.5-10 cm. latis
brcvi-acuminatis obtusis integerrimis utrinque glabris vel prae-
scrtim in nerviis sparsissime adpresso-puberulentis subtus palh-
dioribus basi sensim angustatis acutis, pctiohs 7-15 mm. longis
supra canalyculatis; inflorescentiis in axillariis superioribus
cymosis plus minusve adpresso-sordido-pubescentibus, pedun-
culo usque ad 2 cm. longo; floribus cum pedicello articulatis et
caducis; calyce griseo-tomentoso parvo circiter 2 mm. alto
5-lobato, lobis ovatis obtusis 1 mm. longis; petalo 5 oblongo-
lanceolato 8 mm. longo 2 mm. lato verisimiHter albo utrinque
glabro intus longitudinaliter insigniter unicostato; ovario et
stylo glabro; fructu et seminibus ignotis.
Specimen examined:
Mexico: State of Vera Cruz, Sierra Madre near Miscantla,
August, 1912, C. A. Purpus, 6159 (Mo. Bot. Gard. Herb.)
TYPE.
Specimens of the plant here described were submitted to the
Missouri Botanical Garden for determination by Mr. T. S.
Brandegee who suggested its probable relationship with Mappia,
After a careful study of the material at hand it seems unmis-
takably to belong to the Icacinaceae, but until the fruit is
known its exact position in the family must remain doubtful.
1914]
GREENMAN AND THOMPSON — FLOWERING PLANTS 409
In habit and in the structure of the flower it possesses certain
characters in common with Mappia, Kummeria and Poraqueiha,
but in a combination of the floral characters, particularly in the
free or merely coherent glabrous and strongly ribbed petals,
the broad smooth filaments, elongated anthers, which in cross
section are distinctly x-shaped, and in the single suspended
ovule the plant in question differs from the genera above men-
tioned. Generic rank is therefore given to it and w^e propose
the name Oecopetalum, from okos house and irkaXov petal, in
reference to the httle recesses or pockets formed by the adjacent
petals in which the anthers rest.
Stemodia linearifolia (Morong) Greenman & Thompson,
comb. nov.
Stemodiacra linearifolia Morong, Ann. N. Y. Acad. Sci. 7: 183.
1893.
Stemodia tomentosa (Mill.) Greenman & Thompson, comb.
nov.
Erinus tomentosus Mill. Diet. 1768. [8th ed.]—Herpestes
tomentosa Schlecht. & Cham. Linnsea 5 : 106. IS^O.Stemodia
lanata Ruiz & Pav. in DC. Prodr. 10: 383. 1846; Hemsl. Biol.
Cent.-Am. Bot. 2 : 450. 1882. — Stemodiacra tomentosa 0. Kuntze,
Rev. Gen. 2:466. 1891.
Siphonoglossa Greggii Greenman & Thompson, sp. nov.
Suffruticosa; caulibus erectis vel adscendentibus 0.5-2 dm.
longis subcyhndratis et ssepe quadrisulcatis pubescentibus in
lineis decussatis cum pilis reflexis; foliis oppositis brevipetio-
latis lanceolatis vel obovatis 0.5-2.5 cm. longis 3-7 mm. latis
acutis vel obtusis vel rotundatis integris basi in petiolum grada-
tim angustatis supra glabris subtus paulo pallidioribus juventate
secundum nervos venasque adpresso-puberulentis; floribus in
axillis supernis solitariis sessiHbusque, bracteis subspathulatis;
calyce profunde 5-partito 4 mm. longo, laciniis Hneari-lanceolaf
glabris; corolla 1.5-2 cm. longa bilabiata, labio anteriore horizon-
taliter patenti trilobulato, labio posteriore suberecto emarginato
tubo gracih 9-14 mm. longo extus pubescenti; ovario et style
glabro; capsula circiter 7 mm. longa glabra, seminibus suborbic-
ularibus compressis verrucosis circiter 2 mm. diametro.
Specimens examined:
Mexico : State of Tamaulipas, Matamoras, 7 June, 1847, Dr. J
[Vol. I
410 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Gregg, 915 (Mo. Bot. Gard. Herb.), type; Cervallo, 29 May,
1847, Dr. J. Gregg, 8^5 (Mo. Bot. Gard. Herb.).
Texas: Rio Hondo, Cameron County, September, 1913, Harley
P. Chandler, 7081 (Mo. Bot. Gard. Herb.).
The species here proposed is nearly related to S. Pilosella Torr.
from which it is distinguished by the pubescence of the stem,
namely reflexed hairs disposed in decussating lines, somewhat
narrower leaves, and uniformly shorter fruit.
Siphonoglossa Pilosella Torr. Bot. Mex. Bound. 124. 1859.
This species is well represented in the herbarium of the Mis-
souri Botanical Garden by a suite of more than thirty specimens.
To it should be referred one of Lindheimer's Texas plants,
namely number 1065, collected in 1851, which by clerical error
was distributed as "Ruellia Parryi Gray."
Randia Gaumeri Greenman & Thompson, sp. nov.
Frutex ramosus; caule ramisque cortice griseo glabro tectis;
spinis axillaribus usque ad 1.5 cm. longis divaricatis; foliis
obovatis 0.5-1.5 cm. longis apice plerumque rotundatis integris
basi in petiolum marginatum contractis utrinque glabris vel
supra in nervis ad basin puberulentis ; floribus axillaribus sessi-
libus; calyce toto 1-1.5 mm. longo 4-lobato glabro; lobis triangu-
laribus acutis ciliatis; corolla hypocraterimorpha parva 4-lobata,
tubo cylindraceo circiter 2.5 mm. longo extrinsecus glabro, lobis
contortis ovatis tubo subaequantibus; antheris ad faucem corollas
sessilibus exsertis; ovario biloculari; bacca ignota.
Specimen examined :
Mexico: State of Yucatan, at Izamal, coll. of 1895, Dr. Geo. F,
Gaumer, 589 (Mo. Bot. Gard. Herb.), type.
The divaricately spreading axillary spines, relatively small
obovate leaves, and the minute flowers amply distinguish this
species from all others of the genus. It is with pleasure that
the authors dedicate this new species to Dr. Gaumer, who has
done so much to further our knowledge of the flora of Yucatan.
Randia Purpusii Greenman & Thompson, sp. nov.
Verisimiliter frutex; ramis ramulisque cortice brunneo vel
griseo tectis; spinis ad apices ramorum plerumque quaternis
vel binis, vel rarius nullis, 3-6 mm. longis; foliis lanceolatis vel
obovato-Ianceolatis 1.5-5.5 cm. longis 0.8-2 cm. latis obtusis
1914]
GREENMAN AND THOMPSON — FLOWERING PLANTS 411
»
vel acutis integris basi in petiolum marginatum gradatim angus-
tatis supra hirsutis subtus paulo pallidioribus et subtomentosis;
stipulis triangulari-ovatis utrinque pubescentibus; floribus sessil-
ibus axillaribus terminalibus ; calyce toto 6-7 mm. longo 4-
lobatO; tubo 1.5 mm. longo sericeo, lobis linearibus vel anguste
spathulatis 3-3.5 mm. longis patentibus parce pubescentibus;
corolla hypocraterimorpha profunde 4-Iobata, tubo cylindraceo
fere 1.5 cm. longo extus parce piloso, lobis oblongo-lanceolatis
tubo subsequantibus ; antheris ad faucem coroUse paulum exser-
tis; ovario biloculari, ovulis plurimis; fructu ignoto.
Specimen examined:
Mexico: State of San Luis Potosi, Minas de San Rafael, May,
1911, C. A. Purpus, 6208 (Mo. Bot. Gard. Herb.), type.
Randia truncata Greenman & Thompson, sp. nov. Plate 26.
Frutex erectus 3-4 m. altus ramosus; caule ramisque tereti
cortice griseo tectis juventate parce strigulosis mox glabratis;
spinis 0.5-1 cm. longis binis ad apices ramorum; foliis obovatis
vel spathulatis 0.5-3 cm. longis 0.3-1.7 cm. latis ad apicem
rotundatis obtusis vel submucronato-acutis integris utrinque
glabris basi in petiolum marginatum plus minusve abrupte
contractis; floribus sessilibus axillaribus terminahbus; calyce
toto 1.5-2 mm. longo, limbo cupuliformi truncato; corolla
hypocraterimorpha in sicco atrato, tubo cylindraceo 1-1.5 cm.
longo extus glabro intus sparse piloso, lobis subovatis 4-5 mm.
longis 3-4 mm. latis apice rotundato vel brevissime acuminato;
antheris ad faucem corollae sessilibus semiinclusis; bacca im-
matura globulosa circiter 0.5 cm. diametro.
Specimens examined:
Mexico: State of Yucatan, vicinity of Izamal, coll. of 1895, Dr,
Geo. F. Gaumer, 713, type, and 506 (both in Mo. Bot. Gard.
Herb.); road to Progresso north of Merida, 7 April, 1865,
Schott, 262 (Mo. Bot. Gard. Herb.), distributed as "R,
aculeata."
Co-types of the above species may be looked for in herbaria
under R. xalapensis under which name Dr. Gaumer's material
cited above was distributed. From this species, however, R.
truncata differs in the more obovate outline and the less conspic-
uous veins of the leaf, the somewhat longer and more slender
corolla-tube, and in the smaller truncate calvx.
[Vol. 1
412 ] ANNALS OF THE MISSOURI BOTANICAL GARDEN
Sclerocarpus elongatus (Greenm.) Greenman & Thompson,
comb. nov.
S. Schiedeanus var. elongatus Greenm. Proc. Am. Acad. 32 : 309.
1897.
Herbaceus; caule tcreti ramoso erecto vcl adscendenti 1-1.5
m. alto striato sparse strigoso plus minusve purpm^ascenti basi
lignesccnti; foliis brevipctiolatis trinerviis inferioribus oppositis
superioribus alternis anguste lanceolatis 2.5-13 cm. longis 0.3-
1.5 cm. latis aciiminatis acutis integris vel remote denticulatis
basi in petiolum gradatim angustatis supra tuberculato-hispidis
subtus paulo pallidioribus secundum nervos venasque hirsutis;
inflorescentiis laxe paniculatis, pedunculis gracilibus 0.5-8 cm.
longis strigosis; capitulis 6-8 mm. altis; involucris subcampanu-
latis circiter 5 mm. altis, squamis biseriatis oblongo-lanceolatis
ovatis vel subobovatis extus strigoso-pubescentibus ciliatis
levitcr atratolineatis; flosculis liguliferis 5-8, ligulis oblongig
6-10 mm. longis flavis; floribus disci circiter 30; achseniis matur-
itate obliquis striatis glabris.
Specimens examined:
Mexico: State of Morclos, fields around Cuernavaca, altitude
1585 m., 31 October, 189G, C. G. PriiigU, 6606 (Mo. Bot.
Gard. Herb.), co-type; valley, near Cuantla, altitude 1370
m., 28 October, 1900, C. G. Pringk, 9061 (Mo. Bot. Gard.
Herb.). State of Vera Cruz, Ojapa, 30 June, 1910, C. R.
Orcuit, 5156 (Mo. Bot. Gard. Herb.).
Venezuela: without definite locahty, A. Fcndler, 685 (Mo. Bot.
Gard. Herb.).
A further study of co-type material of this species, supple-
mented by subsequent collections, and a careful comparison of
it with S. Schiedeanus (DC.) Benth. & Hook, f., as represented
by Schiede's number 225 preserved in the herbarium of the
Missouri Botanical Garden and Pringle's number 8338 from the
type locality, shows several important differences between the
species and the plant referred to it as variety elongatus. The
latter has narrowly lanceolate leaves, a much-branched stem,
open inflorescence, and more numerous and smaller heads which
altogether indicate that the plant in question should be regarded
as of equal specific rank rather than a variety of S. Schiedeanus,
hence it is here raised to specific rank and a somewhat amplified
description is appended.
1914]
GREENMAN AND THOMPSON — FLOWERING PLANTS 413
Flaveria longifolia Gray, PI. Fendl. 88. 1849.
Var. subtomentosa Greenman & Thompson, var. nov.
Formse typicse habitu simili; caule plus minusve tomentosoj
foliis lanceolato-attenuatis basi plerumque ampliatis amplexi-
caulibusque utrinque subtomentosis.
Specimens examined:
Mexico : State of San Luis Potosi, Minas de San Rafael, Novem-
ber, 1910, C. A. Purpus, 1^776 (Mo. Bot. Gard. Herb.),
type; Rio Verde, 17 November, 1910, C. Tt. Orcutt, 51^21
(Mo. Bot. Gard. Herb.); Rio Verde, 2-8 June, 1904, Dr.
Edward Palmer, 26 (Mo. Bot. Gard. Herb.).
414
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1. 19141
Explanation of Plate
PLATE 24
Sisyrinchmm ai^gusiisnmxm (Rob. & Grccnm.) Greenm. & Thoinp.
Mexico
From the type number, Pringle No. 4703, in the Herbarium of the Missouri
Botanical Garden.
Ann. Mo. Box. Gard.. Vol U 1914
Pl^TE 24
GREENMAN AND THOMPSON— DIAGNOSEvS OF FLOWERING PLANTS
COCKAYNE, BOSTON.
[Vol, 1, 1914
416 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 25
Oecopetalum mexicanum Greenm. & Thomp.
Fig8. 1 and 2j flowering branches; 3, flower; 4, inner face of petal; 5 and 6, front
and side view of stamen; 7, longitudinal section of pistil; 8, diagrammatic cross-
section of flower bud; 9, diagrammatic cross-section of anther before and after
dehiscence.
L
Mexico
From the type specimen, Purpus No. 6159, in the Herbarium of the Missouri
Botanical Garden.
1
i
4
Ann. Mo. Bot. Gard., Vol 1. 1914
Plate 25
I
GREENMAN and THOMPSON— DIAGNOSES OF FLOWERING PLANTS
COCKAYNE:. BOSTOrJ,
.-lo [Vol. I. 1911
415 ANNALS OF THE MISSOURI BOTANICAL GARDEN
Explanation of Plate
PLATE 26
Randia truncata Grccnm. & Thomp.
Mexico
From the type specimen, Gaumer No. 713, in the Herbarium of the Missouri
Botanical Garden.
Ann. Mo. Bot. Gard., Vol 1, 1914
Plate: 26
GREENMAN AND THOMPSOxN—DlAGNOvSES OF FLOWERING PLANTS
COCK A > NE, c-OStON.
I ■
ENZYME ACTION IN FUCUS VESICULOSUS L.
B. M. DUGGAR
Physiologist to the Missouri Botanical Garden^ in Charge of Graduate Laboratory
Professor of Plant Physiology in the Henry Shaw School of Botany of
Washington University
AND A. R. DAVIS
Rufus J. Lackland Fellow in the Henry Shaio School of Botany of
Washington University
Little is known regarding the metabolism of the Fucaceoe.
The chemical nature of the chief accumulation products has not
yet been sufficiently investigated. In fact, prior to 1905 very
little work of importance had been contributed on the products
of any group of the marine algse. Even the chemical deter-
mination of the carbohydrates, for example, in some of the larger
groups of algse, afforded no suggestion as to the nature of these
products. More activity in this general field of work has been
manifest, however, since the date referred to. Diverse views
prevailed regarding the nature of the various granules which had
been long detected microscopically. In the earlier literature
Hansteen's ('92, '00) opinion has generally dominated, by which
it was claimed that the granular bodies of the cell — and par-
ticularly the larger vesicular forms — contain fucosan, a carbo-
hydrate, which was considered the first visible product of
photosynthesis. On the other hand, Crato ('92, '93) main-
tained, from microchemical reactions, that the larger vesicles,
physodes as he called them, contained phloroglucin, or some
derivative of this body. Miither and Tollens ('04) found a
methylpentosan in Fucus and Laminaria, while Koenig and
Bettels ('05) among others found glucose and fructose, as well
as pentoses and methyl pentoses, in Laminaria after hydrolysis.
Swartz ('11) gives an extensive summary of the previous^work
on carbohydrate occurrence in the algse, and contributes much
data on the digestion of the hemicelluloses, but she studied no
brown algse. The existence of reducing sugars in Fucacece was
clearly shown by Tihomirow ('10). Recently the carbohydrates
have been more completely investigated by Kylin ('12, '13).
Nevertheless, much remains to be done on these products, while
Ann. Mo. Bot. Gabd., Yol. 1, 1914
(419)
1
I
i
1
[Vol. 1
420 ANNALS OF THE MISSOURI BOTANICAL GARDEN
the proteins (aside from agar agar and related compounds) and
other organic substances are scarcely known.
In view of the very considerable data on the carbohydrate
metabolism in higher plants, it seems particuharly desirable to
investigate further this relation in the brown algse. Moreover,
no general study having been made, as far as we could learn, of
the enzymes of the Fucacece, it seemed possible that a determi-
nation of the more characteristic enzymes, and of their distri-
bution in Fucus, might lead to a better comprehension of the
nature of the metabolism of these plants. Accordingly, during
the summers of 1913-14 we have made an examination of
Fiicus vesiculosus with respect to its enzyme content.
In preparing the Fucus material for study we have followed
several of the customary methods which have been found
satisfactory in yielding enzymes of a high degree of efficiency.
Since our results with Fucus have been so generally negative
with respect to the presence of the commoner enzymes of plant
metabolism, it may be well to indicate briefly how the material
was handled. The Fucus plants were obtained in quantity,
apparently in a condition of active growth, and the material
was carefully picked over to avoid the contamination of attached
animals and smaller alga?, then washed, and finally treated by
one of several methods. Some of it was hung in a shaded, warm
room until quite dry and brittle, then ground in a mill to an
extremely fine powder, the latter being preserved in dry bottles
for extraction, as subsequently indicated. For other phases of
the work the plants fresh from the water were ground almost to
a pulp in a meat grinder, sometimes passing the material twice
or oftener through the machine. In some cases this fresh pulp,
further comminuted in a mortar, or an extract from it, was used
directly, while in other cases an alcohol-acetone dry preparation
was made from it — the latter by treating alternately with 95 p(T
cent alcohol (15 minutes) and acetone (5-10 minutes) until
practically dehydrated, with a final brief treatment with absolute
alcohol or ether, when the material was spread out on filter
paper to dry. The alcohol-acetone material was thoroughly
pulverized in a mortar for further use.
In the preparation of extracts the dry material was treated
with distilled water (usually 10 parts of water to 1 part of
19U]
DUGGAR AND DAVIS — FUCUS VESICULOSUS 421
material), or in some cases with sea- water, using commonly 20
per cent alcohol or 2-3 per cent toluene as a preservative. In
general, toluene has proved the most satisfactory antiseptic.
The filtered extract was then precipitated with 95 per cent
alcohol, the precipitate caught on a filter, washed with alcohol
and dried. In a few cases the extract was used direct, and in
certain respects the common practices were variously modified
in the hope of detecting some simple explanation of the large
number of negative results.
The hydrolytic experiments were carried out in small Erlen-
meyer flasks or test-tubes, and always in dupHcate or tripli-
cate. In addition, nearly every series was repeated once or
oftener. A special effort was made to determine the presence
of carbohydrases, and for this purpose weak solutions, usually
0.5 per cent, of starch, glycogen, dextrin, saccharose, maltose,
and lactose were employed in numerous tests. No reduction,
or no change in the reducing value of the substrate, by the
Fehling method, was found in any case in our final experiments,
although in some cases a relatively large quantity of the sup-
posedly enzyme-containing material was used. We found it
purify the best dextrin obtainable by
with 95 Der cent alcohol from
In the
k
preliminary experiments, and chiefly with one preparation,
traces of reduction were found with glycogen, but in many later
experiments this finding was not confirmed.
Owing to the consistently negative results with these carbo-
hydrates it seemed possible that there might be an adjustment
of enzyme action in Fucus such that a relation of the mineral
salts, as in sea water, might be requisite for highest action.
Consequently the enzyme solution in one large scries of experi-
ments was diluted with double strength sea- water; in another
case the material was extracted with sea- water; and finally,
fresh material was used, making with it a diffusion in sea water.
In every instance the result was negative.
Another possibility then suggested itself, namely, that the
presence of certain inhibiting substances might account for the
absence of hydrolytic change. Accordingly, the effect of the
Fucus material on the activity of taka diastase was determined
in this way: To 10 grams of ground fresh material 100 cc. of
5
t
[Vol. 1
422 ] ANNALS OF THE MISSOURI BOTANICAL GARDEN
water and 1 gram of taka diastase were added, this being per-
mitted to stand for 6 hours, as in extraction, and the filtrate
from this extraction was tested upon starch solution. The
results were positive, indicating that no free substances were
present which could inhibit diastase action. In another test
1000 grams of Fucus material were divided into two lots of 500
grams each. To one of these, 5 grams of commercial malt
diastase were added, and both were then treated by the alcohol-
acetone method, and subsequently extracted and precipitated
in the usual way. The material to which diastase had been
added gave positive tests for the hydrolysis of carbohydrates
in an extensive series with dextrin, glycogen, saccharose, and
laminarin; but a solution of the precipitate from the lot receiving
no diastase produced no changes in these substrates. These
experiments included controls of several lands. With every
substrate, boiled material was also used, and it is interesting
to note that the "enzyme" material increased in reducing power
with boiling.
The tests referred to in the previous paragraph seemed all the
more important inasmuch as the Fucus material had been found
to be strongly acid, and it seemed possible that this acidity
alone might prove an injurious factor. From the experiments
just mentioned it is seen, however, that acidity could scarcely
have been an important consideration. A quantitative deter-
mination of the acidity was nevertheless made, by titration with
NaOH, and it was found to be about .0565 N HCl. There is a
slight increase in the acidity, if the pulp is permitted to remain
in water 12 hours.
Owing to the determination by many, as, for example, Mlither
and Tollcns ('04), Kylin ('13), Swartz ('11), and others of the
presence of hemicelluloses, especially pentosans, in the marine
algai, and, further, since the commoner carbohydrate enzymes
had not been identified by us, it seemed desirable to examine the
material for pentosanasc. The most available pentosan was
that of cherry gum, accordingly this material in fresh condition
was obtained and utiUzcd in many tests with the Fucus prepara-
tion, the flasks being maintained at temperatures ranging from
27-40° C. Although the experiments were permitted to run for
a period of several days, no reduction above the amount found
1014]
DUGGAR AND DAVIS — FUCUS VESICULOSUS 423
in the controls was obtained, and certainly no pentosanase active
on this material could be assumed to occur abundantly in Fucus
tissues.
Only one series of tests has been made to identify cellulase
in the material here reported upon, and the results are presented
with much reserve. Precipitated cellulose, prepared from filter
paper, was employed, and the experiments were conducted at
40° C» The indications were that slight cellulase activity may
occur.
By means of the action of the alcohol-acetone preparation
upon a 4 per cent olive oil-casein emulsion, the lipolytic activity
was investigated in the usual way. With the emulsion used
alcohol is most serviceable as a preservative. In the tests
referred to there was no indication of hydrolysis after one week;
so the preparations were permitted to stand for two months,
but still without change. That the conditions in the above
case were otherwise favorable for lipolytic action is shown by the
fact that the same substrate yielded with an alga of another
family a decidedly positive test in two days. Several series of
experiments were likewise carried out for the determination of
esterases. With methyl acetate, ethyl acetate, and ethyl but}^-
rate the Fucus material produced no change, irrespective of the
concentrations employed.
In some of our preliminary experiments it had appeared that
urease was present, but a careful investigation of this point
demonstrated an error in the earlier results, and no amidases
were discovered through the action upon 0.5 per cent solutions
of urea, acetamid, methylamine, asparagin, diphenylamine, and
acetanilid. In these experiments NH3 determinations were
made according to the method of Folin.
No liquefaction of gelatin or of agar occurred during a ten-day
interval in a large series of test-tubes arranged with these two
substrates. In the different tests these media were made
neutral, alkaline, and slightly acid. In the neutral and slightly
acid tubes no observable change occurred; but in those tubes
containing a higher percentage of acid — both in those contain-
ing the Fucus preparation and in the controls — general lique-
faction occurred. It is obvious, therefore, that these gel-
forming proteins are not noticeably affected by any enzymes
424 ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
occurring in the Fucus material. More extensive scries of tests
were arranged to determine the presence of proteinases which
might act upon some more widely distributed native proteins,
such as albumin, casein, and legumin. No tests were made to
determine the transformation of these bodies into proteoses or
peptones, but the formaldehyde method of determining amino
acids was employed, and in no case had any" transformation of
these substances proceeded to the amino acid stage.
Glucose, levulosc, and galactose were used in two series of
experiments designed to determine the presence of zymase in
the alcohol-acetone Fucus powder. No sufficient evidence,
however, of the occurrence of this enzyme was obtained even
when the most delicate tests were employed to determine the
liberation of CO2. The action of Fucus extract from the
alcohol-acetone preparation upon tannin was tested by means of
quadruplicate experiments. Two concentrations of tannin
were used, 1 per cent and 2.5 per cent. The determinations
were made by means of Jean's iodine method, but in no case did
the flasks receiving the Fucus extract exhibit hj^drolysis greater
than that shown by the controls. Neither prepared nor fresh
Fucus material gave sufficient evidence of oxidase or peroxidase
action to be considered positive. Negative results were obtained
both by the direct method with gum guaiacum, and by the in-
direct method, in which the reagent mentioned is used with
hydrogen peroxide, and apparently acidity is not a determin-
ing factor. The use of benzidine seemed to indicate oxidase
activity, but it has been clearly shown that the ease with
which this reagent undergoes ''spontaneous" oxidation in boiled
solutions necessitates caution in using it as a test of oxidase
activity. Tests for catalase by the usual method, evolution of
oxygen on the addition of hydrogen peroxide, have clearly in-
dicated that this enzyme is widespread in Fucus. It should be
noted that the findings with respect to oxidase and catalase
activity are in agreement with those of Atkins ('14). Catalase
was very generally identified by him in the algae, but evidence
of oxidase in the Fucaceoe was obtained only with benzidine as a
reagent.
The unexpectedly negative character of the experimental
I
work here briefly outlined prompted us to make many repeti-
i
K
1014]
DUGGAH AND DAVIS — FUCUS VESICULOSUS 425
tions of experiments and minor modifications in technique not
referred to in this preliminary account. The nature of the
results, furthermore, made it seem desirable that a much moi*e
general study be made of the abundance and distribution of the
enzymes in the various families of the marine algse, and such an
investigation is now in progress by one of us.
It would seem idle to attempt here an explanation of the nega-
tive results obtained, yet two or three possibilities have occurred
to us which may be mentioned. The conditions of life of the
Fucacece, especially the temperature relation, make it possible
that metabolic chang
If
this is the case, it might be assumed that the commoner metabo-
lic enzymes might be present in such small quantity that an
indication of their presence would not be apparent by utilizing
the methods ordinarily employed. The very fact that the
capacity for food accumulation, that is to say, the ''storage" of
food materials, has not become highly developed in these forms
suggests that the usual enzymes might not be found in abun-
dance. Nevertheless, if such is the case, it may be pointed out
that the present methods of enzyme work are very inadequate
when applied to metabolic processes in general dealing with the
transformation of products which do not accumulate in some
quantity in the cell. In this connection attention may be
drawn to Arbor's ('01) observation on the slow rate of trans-
formation of starch in the thallus of Ulva latissima, where a
darkening period of from three to five weeks was required for
the disappearance of this product.
The other possibility which has suggested itself is that in the
cells of the Fucacem there may occur inhibiting substances which
upon the death of the cell may form with the enzymes com-
pounds from which the ferments cannot be again recovered.
We have no evidence of the existence of any such bodies. Fur-
ther investigation of Fucus and related algse should perhaps
throw some light upon the negative evidence produced by our
extensive data.
Graduate Laboratory, Missouri Botanical Garden,
«
.o^ tVoL. 1, 1914]
4zb ANNALS OF THE MISSOURI BOTANICAL GARDEN
Literature Cited
4
Arl)cr, E. A. N. (^01). On the effects of salts on the assimilation of carbon dioxide
in Ulva latissima. Ann. Bot. 15:39-69. 1901.
Atkins, W. R. G. ('14). Oxydases and their inhibitors in plant tissues. Tart III:
The looahzation of oxydases and catalase in some marine algse. Scientif. Proc.
Roy. Dul)lin Soc, N. S. 14: 199-20G. 1914.
Crato, E. ('92). Die Physode, ein Organ deg Zcllcnlcibcs. Ber. d. dcut. bot. Ges.
10:295-302. pi 18, 1892.
, C93). IJcbcr die Hansteen'schcn Fucosankorncr. Ibid. 11: 235-241.
1893.
Hanstccn, B. ('92). Studien zur Anatomie und Physiologic dor Fucoidcen. Jahrb.
f. wisa. Bot. 24:317-362. pi 7-10. 1892.
, ('00). Ueber das Fucosan als erstes schcinbarcs Product dcr Kohlensaurc-
assimilation bei den Fucoidcen, Ibid. 35:611-625. pi I4. 1900.
Koenig and Bcttels ('05). Die Kohlenhydratc dcr Mecrcsalgen und daraus herge-
stclltcr Erzeugnissc. Zeitschr. f. Unters. d. Nahrungs- u. Genussmittel 10: 457-
473. 1905.
Kylin, H. (42). Ueber die Inhaltskorper dcr Fucoldecn. Arkiv f. Bot. 11^:1-26.
pi L 1912.
CI 3). Zur Biochemie dcr Mecrcsalgen. Iloppe-Scyler's Zeitschr. f.
I
I
physiol. Chem. 83: 171-197. 1913.
Mather and Tollcns ('04). tjber die Troducte der Hydrolyze von Reetang (Fucus),
Laminaria, und Carragheenmoos. Zeitschr, d. Ver. d. deut. Zucker Tnd,
54:59. 1904. [Cited by Swartz.l
Swartz, M. D. ('11). Nutrition investigations on the rarbohydrates of lichens,
algaj, and related substances. Trans. Conn. Acad. Arts and Sci. 16: 247-382.
1911.
Tihomirowj W. A. ('10). Sur la valcur de la reaction microchimiquc de la pli6nyl-
hydrazine: pour la constatation du sucre dans lea tissus des plantea. Ann.
Jardin Bot. de Buitenzorg, Sup{)I. 3=: 537-582. pi. 13-15. 1910.
4
GENERAL INDEX
New scientific names of plants and the final members of new combinations are
printed in bold face type; synonyms and page numbers having reference to figures
or plates, in italic; and previously published scientific names and all other matter,
in ordinary type.
\
A
Acid, oxalic, produced by Sclerotinia
cinerea, 319
Acidity: The effect of certain conditions
upon the, of tomato fruits, 229; of
lum fruits, 317; relation of growth of
clerotinia cinerea to, 318
Agaricacese, 197
Agaricus beiulinus, 146; quercinus, 145
Air, The identification of the most char-
acteristic salivary organism, and its
relation to the pollution of, 47
Air-sampling apparatus, bacteriological,
SO; construction and use of, 66
albido-brunnea (Thelephora), 214, 216,
228
albo-violascens (Cyphella), 364
Aleurodiscus, 198
Algae: A contribution to our knowledge
of the relation of certain species of
grass-green, to elementary nitrogen,
157; Some pur^ culture methods in the,
23; Indications, regarding the source
of combined nitrogen for Ulva Lactuca,
229
angustata (Thelephora), 205
anomala (Solenia), 373^ var. arhiculariSj
373
Anthericum Chandleri, 405 ; stenocar-
pum, 400
anthocephala
(Thelephora), 202, 203,
/^
arachnoidea (Cyphella), 363
Arrhenia, 344
artocreas (Michenera), 197
Asterostroma, 198
Augustinii (Heterothecium),
B
381
Banana? (Cyphella), 379
biennis (Thelephora), 215
Boletus abietinus, 91 ; adusluSy 102 ;
applanatuSj 137; arculariuSj 107; betu-
linuSj 104; brumalis, 107; caesius^ 96;
cinnabarinuSf 116; cinnamomeus, 123;
Ann. Mo. Box. Gard., Vol. I, 1914
conchatuSj 132; conchifer, 93; con-
fragosuSj 144; cuticidaris^ 1 IS; dis^
tortus, 105 ; dryadeus^ 119 ; elegans^
110; fomentariuSj 136; fraxineus, 130;
frondosuSf 112; fidvus, 133; fiimosus^
103; gilviis, 117; giganteiis, 113; grave-
olens, 131; hirsutus, 93; hispidus, 119;
igniariiiSy 135; luciduSj 123; nummu^
larius, 110; perenniSj 122; Pint, 142;
pinicolaj 130; pubescenSj 94; radiatus,
118; resinosuSy 116; sanguineus^ 115;
spumeusj 99; sqicamosusj 109; suxive^
olenSj 140; sulphureus, 114; tulipifera,
152; umhellatuSj 112; unicolor, 143;
versicolor, 91
Bordeaux mixture, effect of, on trans-
piration, 12, 351
borealis (Craterellus), 357, 382 ^
Botrydiopsis sp., isolation of, in pure
culture, 38
Botrydium granulatum, isolation of,
in pure culture, 38
brevipes {Cantharellus), 329
Burt, E. A. The Thelephoracese of
North America, I, 185; II, 327; III,
357
c
Cacalia prenanthoid-'s, 271 ; runcinata^
272; Toluccana, 271
caespitulans (Thelephora); 204, 340
calceum (Corticium), 187
calyculus (Craterellus), 338
campanula {Peziza), 360
canadensis (Cantharellus), 345
canadensis (Craterellus), 345
Candida {Cyphella), 377
Cantharellus, 197; brevipes, 329;
densis, 345; floccosus, 345;
362
Cantharellus (Craterellus), 330, 332, 346
capuh (Cyphella), 366, 382
caricina (Cyphella), 366, 382
caryophyllea (Thelephora), 207, 209, 22(3
Castor bean leaves, use of, in potometer
X
cana-
Icevis,
experiments, 7, 11
(427)
428
ANNALS OF THE MISSOrRI BOTANICAL GARDEN
[Vol. 1
Cellulose, methodsof preparation of, 305
Chlamydomonas i)isiforniis forma minor,
isolation of, in pure culture, 32; rela-
tion of, to elementary nitrogen, 178,
179, 181
Chlorascens (Ilcterobasidium), 197
Chlorella ep., and C. vulp;aris, isolation
of, in pure culture, 33; relation of, to
elementary nitrogen, 178, 179, 181
Chlorococcum humicola, isolation of,
in pure culture, 35; relation of, to
elementary nitrogen, 178, 179^ 181
cinereo-fusca (CjT)hclla), 377
cinereo'fusca {Peziza), 377
Cladodcrris, 198
Clavaria pistillaria, 342
clavatus (Cratcrcllus), 329, 345, SI^G
conjluens {Craterellus) ^ 332
conglobata (Cypliella), 375, 383
Coniophora, 198, 199
convoluta (Cy{)hella), 380
Cqoley, J. S. : A study of the physiolog-
ical relations of Sclerotiiua cinerea
(Bon.) Schroter, 291; Duggar, B. M.,
and, The effect of surface films and
dusts on the rate of transpkation, 1,
The effects of surface fihns on the rate
of transpiration: exi)criments with
potted potatoes, 351
Cora, 199
corbiformis (Thelephora), 211
Coriolus Lloydii^ 95; 7iigro)narginaluSf
cornucopioides
350
(Craterellus), 327
cornucopioides (Thelephora), 212, 346
corrugis {Cralerdlm) , 340
Corticium, 190, 191, 192, 193, 197, 198.
199; calceum, 187; lactescens, latex in,
194; lacteum, 187; Sambuci, 191; sub-
gigantcum, 197; vagum, 191.
Craterellus, 190, 196, 197, "^7; borealis,
357, 3S2; calyculus, :.j6; canadensis,
345; Cantharcllus, 330, 332, 346;
clavatus, 329, 345 S46; conjluens,
332; cornucopioi^les, 327, 333, 350;
corrugis, 340; c^ispus, 338; delitescens,
339, 350; dilatus, 343, 350; dubius,
335; Humphreyi, 344, 350; lakntius,
330; lutescens, 336, 350; Pogonali, 362,
SS2j maravsmioides, 345; ochrosporus,
327, 334, 350; ocreatus, 334; odoratus,
327, 331, 340, 348; palmatus, 342,
350; pistillaris, 327, 341, 348, 350;
Pogonaii, 362, 382; pulverulcntua, 345;
roseus, 332; sinuosus, 337; spathu-
larius, 345; taxophilus, 339, 360; uni-
color, 327, 340, 341, 342, 348
criepus (Craterellus), 338
cupuUeformis (Cypholla), 369, 382
Cupressi (Cyphella), 380
cuticularis (Tlielephora), 216, SS8
Cyclomyces, 82, 147; Grcenci, 147
sub-
tex-
365,
365,
Cymatella, 345
Cyphella, 197, 358; albo-violascens, 3C4;
arachnoidea, 363; Banana*, 379; Can-
dida, 377; capula, 366, 38S; caricina,
366, 382; cinereo-fusca, 377; conglo-
bata, 375, 382; convoluta, 380; Cup-
ressi, 380; cupukrformis, 369, 382;
digitalis, 358; fasciculata, 373, 376, 382;
filicicola, 379; fraxinicola, 3(58; j'ulva,
373; fumosa, 376, 383; furcala, 373;
?raleata, 302, 382; glubosa, 367; grisoo-
paUida, 3G9; laita, 361; Langloisii,
368, 382; mellea, 372, 382; minutis-
sima, 367, 368, 382; musuicola, 380;
muscigena, 362, 363, 383; Palmarum,
377; Pcckii, 377; perexigua, 378; pezi-
zoides, 364, 365, 378; porrigens, 368,
382; punctiformis, 367; Ravenelii, 371,
372, 373, 382; Ravenelii Sacc, 373;
Saccardoif 373; subcyanea, 380;
gelatinosa, 370; sulphurea, 360;
cnsis, 371, 373, 382; Tilia?, 364,
382; trachycha)ta, 379; villosa,
382
D
Daedaloa, 82, 143; ambigua, 144; cou-
fragosa, 144j mollis, 141; paUido-fvIva,
146; quercma, 145; sminaria, 147;
unicolor, 143
Davis, A. R., Duggar, B. M. and.
Enzyme action in Fucus vegiculosus.
419
delitescens (Craterellus), 339, S50
Dendrocladium, 199
dentosa (Thelephora), 224, 346
digitalis (Cyphella), 358
dilatus (Craterellus), 343, 350
dubius (Craterellus), 335
Duggar, B. M.: and Coolcy, J. S., The
effect of surface films and dusts on the
rate of transpiration, 1, The effects of
surface fihns on the rate of transpira-
tion: experiments with potted pota-
toes, 351; and Davis, A. R., Enzyme
action in I'ucus vcsioulosus, 419; and
Merrill, M. C, The effect of certain
conditions upon the acidity of tomato
fruits, 229
Dusts, the effect of surface fihns and, on
the rate of transpiration, 1
E
Enslinia pocula, 100
Enzymes; cellulase in Sclerotinia cinerea
and Penicillium expansum, 305; pec-
tinase in Sclerotinia cinerea and Pen-
icillium expansum, 312
Ercchthitcs runcinaia, 271, 272
Erinxis tornenlosuSj 409
Eu-Thelephorea?, 197
Exobaaidium, 193, 197
■
y
^
f
1914]
ANNALS OF TPIE MISSOURI BOTANICAL GARDEN
429
F
fascJculata (Cypliella), 373, 376, 382
Favolus, 82, 148 ; canadensis, 148 ;
ohiensis, 148; rliipidium, 148; striatu-
lus, 148
filicicola (Cyphella), 379
Films and dusts, the effect of surface, on
the rate of transpiration, 1
Films, the effect of surface, on the rate
of transpiration : experiments with
potted potatoes, 351
fimbriata (Thelephora) , 222, S26
Flaveria longifolia, 413, var. subtomen-
tosa, 413
Flowering plants, Diagnoses of, chiefly
from the southwestern United States
and Mexico, 405
Fomes, 82, 126; applanatus, 130, 135,
136, 137, 138; carncus, 131; conchatus,
132; connatus, 129; Everhartii, 134,
135; fomentarius, 135, 136, 137; frax-
ineus, 129, 130; fraxinophilus,^ 129;
fulvus, 133; graveolcns, 131; igniarius,
134, 135; lobatus, 137; nigricans, 134,
135, 136; ohiensis, 128; pinicola, 130;
populimiSj 130; ribis, 133; rimosus,
133, cause of trunk disease of mesquite,
248; roseus, 131; salicinns, 133;
scutellatus, 128; supinuSf 133
Foster, G. L. Indications regarding the
source of combined nitrogen for Ulva
Lactuca, 229
fraxinicola (Cyphella), 368
Fucus vesiculosus, Enzyme action in,
419
fulva (Cyphella) f 373
fumosa (Cyphella), 376, 382
Fungous and host cells, A method for the
¥ differential staining of, 241.
furcata {Cyphella) j 373
griseo-pallida (Cyphella), 369
griseozonata (Thelephora), 221, 2S8
G
galeata (Cyphella), 362, 382
Ganoderma sessile^ 123 ; subperforatuniy
123
Gates, "R. R. A Texan species of Mega-
I pterium, 401; Some Oenotheras from
Cheshire and Lancashire, 383
globosa (Cyphella), 367
Gloeocystidium, 193
Gloeopeniophora, 193
Gloeoporus, 82, 149; conchoides, 149
Grandinia, 196
Greenman, J. M.: Descriptions of North
American Senecioneae, 263; and Thomp-
son, C. H., Diagnoses of flowering
plants, chiefly from the southwestern
United States and Mexico, 405
Grifola poripeSj 111; rainosissima^ 112;
Sumstineij 113
H
Herpestes lomenlosay 409
Heterobasidium, 197 ; chlorasccns,
Ileterothecium Augustinii, 381
Hirneolina, 197, 198
hiscens (Thelephora), 207
Humphrey! (Craterellus), 344, 360
Hydnacese, 196
Hymcnochaetc, 193,
Hymeno-licheng, 199
Hyphomycetes, 194
Hypochnaccse, 190
Hypochnus, 190, 193, 19G, 198, 199
ilypolyssus, 198
197
198
I
Infection and penetration of plums by
Sclerotinia emerea, 297
Inonolus lexanus^ 247
Introduction, i
intybacea (Thelephora), 217, 220, 228 ^
Irpex, 82, 151; cinnamomeus, 152; fari-
naceus, 152, 153; mollis, 152; tulipi-
fera, 152
K
Kirchnericlla sp., isolation of, in pure
culture, 34; relation of, to elementary
nitrogen, 178, 179, 181
Kummeria, 409
L
Lachnocladium, 196, 198, 199
laciniata (Thelephora), 219
ladnialum (Stereiim), 219
lactescens (Corticium), 194
lacteum (Corticium), 187
Iseta (Cyphella), 361
loevis (Cantharellus)f 362
Langloisii (Cyphella), 368, 382
lateritius (Craterellus) j 330
Lenzitcs, 82, 145; bctulina, 146; Cra^
taegif 144; Jlaccida, 146; protractus,
destruction of sa})-wood of mesquita
by, 248; saepiaria, 147; vialis, 146
Lilac, A trunk disease of the, 253
Lloydella, 192
lutesccns (Craterellus), 336, 350
lutosa (Thelephora), 216, 346
M
magnispora (Thelephora), 211, 226
Mappia, 408
marasmioides (Craterellus), 345
N
430
ANNALS OF THE MISSOURI BOTANICAL GARDEN
[Vol. 1
Mcj2;aptcrium, A Texan species of, 401;
argyrophyllum, 401, ^04, vax. retusi-
folium, 401, 404; Frcmontii, 402;
macrocarpunij 402; missourienHis, 402,
403
mellea (Cyphella), 372, 3S2
Merrill, M. C, Du^jz;ar, B. M., and. The
effect of certain conditions upon the
Rcidity of tomato fruits, 229
Merulius, 82, 150, 197; lacrymans, 151;
rubellus, 150; trenicUosus, 151; iremel-
losuSj 151
Mescjuite, Two trunk diseases of the, 243
Michenera, 197; artocreas, 197
Michejicri {Slereum)^ 214
Microcoleus sp., isolation of, in pure cul-
ture, 39
minutissima (Cyphella), 3G7, 308, SSS
Moore, G. T. Foreword, i
Mucronoporus Evcrhartiiy 134
multipartita (Thelephora), 205, 208, SS6
musa^cola (Cyphella), 380
museigena (Cyphella), 3G2, 38S
Mycobonla, 198
N
Navicula sp., isolation of, in pure culture,
39
Nitrogen: A contribution to our knowl-
edge of the relation of certain species
of grass-green algffi to elementary,
157; Indications rejz;arding the source
of combined, for tUva Lactuca, 229
Nolte, A. G. The identification of the
most characteristic salivary organism,
and its relation to the pollution of air,
47
obovata (Xylaria), 225
ochrosporus (Craterellus), 334, 350
ocreatus (Craterellus) 334, 335
odoratus (Craterelhis), 331, S46, 348
odorifera {Thelephoru) ^ 215
Oecopetalum, 408; mexicanum, 408, 4^6
Oedogonium, isolation of zoospores free
from bacteria in, 36
Oenothera Lamarckiana, 393, 4^0; macro-
carpn, 402; multiflora, 386, 396; var.
elliptica, 387, 3nn^ SOS; rubrinervoides,
389, 396, 39S; rubritincta, 391, 398,
4OO; tardiflora, 391, 4OO
Oenotheras, Some, from Cheshire and
Lancashire, 383
Ohio, The Polyporacea? of, 81
Oscillatoria sp., isolation of, in pure cul-
ture, 39
Overholts, L. 0. The Polyporacea? of
Ohio, 81
P
palmata (Tliclephora), 201, S26
■pabnata nmericana {Thelephora) , 201
Palmarum (Cyphella), 377
palmatus (Craterellus), 342, 530
pannosa (Thelephora), 207
Peckii (Cyphella), 377
Penicilhum expansum, physiological
studies in, 308, 315, 322
Peniopliora, 190, 191, 192, 193, 198, 199
Peniophorella, 193
perexigua (Cyphella), 378
Permeability of collodion membranes,
40
perplexa (Thelephora), 223, 346
P(v.iza campamda^ 3()(); Tilia% 364
pezizoides (Cyphella), 364, 365, 378
pistillaris (Clavaria), 342
pistillaris (Craterellus), 341, 348, 350
Pleurococcus vulgaris, isolation of, in
pure cultiHT, 34
Pogonati {Craterellus), 362, 382
Polyporaceie, 197
Polyporacea) of Ohio, The, 81; index to
species of, 153; key to genera of, 85
Polyporus, 82, 80: abietinus, 91, 92;
ahorUvus, 105; adustus, 102, 103, 104;
alhelhiSj{)7] anax, 113, 114; applanatns^
137; arcidariformiSy 107; arcuhirius,
107; hadius, 126; Borkeleyi, 113;
betuhnus, 104; biformis, 95; borealis,
100; brumalis, 107; caesius, 90, 97;
carneuSf 131; castanophilus, 115; chion-
c\\f^,*d7;ci7ici7matHs, 114; cinnabarinus,
116; cinnamomeus, 123; circinatus,
121, 122, 123; conchalus, 132; conchifer,
93 ; conglobalus, 131 ; connalus Fries,
129; connatus Schw., 122; connalus
Weinm., 129; cristalus, 111; Curtisii,
125; cuticularis, 117, 118, 119; delec-
tans, 99; dichrouSy 150; distortus, 105;
dryadeus, 119, 120; dryophilus, 119,
120; dualis, 121; elegans, 110; endocro^
cinus, 119; fibula, 95; fissiis, 109;
flavovirens, 111; foeicola, 122, 123;
fo7ne7itariuSy 13G]fragilis, 126; fragrans,
102, 10^; fraxineus, 130; fraxinophilus,
129; frondosus, 112, 113, 114, 126;
fulims, 133; fumosus, 102, 103; galac-
tinus, 97, 98, 141; giganteus, 113, 126;
gilvus, 117, 118, 120; guttulatus, 100;
hirsutulus, 9 1 , 92 ; hirsut us, 93 ; hispidus,
119; hypococcineus, 115] igniarius, 135;
imimtuSj 98; intyhaccus^ 120; isidioideSf
117; lacteus, 97; Icnius, 126; Icuco-
melas, 126; leucophmis, 137; lobatus,
137; Lloydii, 95; lucidus, 123; macula^
his, 100; mollitiscidus, 95, 96; Morgani,
110; nidulans, 117; nigricans, 136;
obesus, 121, 123; obtusus, 100; ovinns,
126; pargamcnus, 92; parvidus, 122;
pennsylvanicus, 108; percnnis, 122,
1914]
ANNALS OF THE MISSOURI BOTANICAL GARDEN
431
123; perplexus, 118; phaeoxarUhuSj 126;
picipos, 109, 110; Pilotae, 115; pini-
cola, 130; pocula, 106; populinus, 141;
puherula, 103; pubescens, 94, 96, var.
Grayii, 95; radiatug, 117, 118, 119;
radicatU8, 110; reniformiSj 137j resia-
0S113, 116; rimosus, 133; robiaiophila,
104; Rostkowii, 109; rujescens, 106;
Banguineus, 115, 116; Schweinitzii,
120; scutellatas, 128; semipileatus, 96;
serialis, 139; Spraguei, 101; spumcug,
99; squamo3U3, 109; subsericeuSy 123;
Sidlivantiif 94; sulphureu3, 114; texa-
nus, 247, cause of truak disease of
mesquite, 246, 350, 252; tomeatosus,
121; umbellatus, 112, 113; varius, 110;
velutinuSj 93, 95; versicolor, 91, 93,
cause of trunk disease of lilac, 253,
260, 262; virgineus, 94; volvatus, 105;
zoaalis, 101; zonatus, 91
Polystictus, 82; Lindheimeri, deatruc-
tion of sap-wood of mesquite by, 248;
ohesu9, 121; proliferus, 123
Poraquciba, 409
Poria, 82
porrigeas (Cyphella), 368, 3S2
Potato plants, transpiration of, 351
Potometer, 22; experiments with, 7
Prosopis glandulosa, trunk diseases of,
243
Protosiplion botryoides, isolation of, in
pure culture, 35; relation of, to elemen-
tary nitrogen, 178, 179, 181
pulverulentus (Craterellus), 345
punctiformis (Cyphella), 367
Pure culture methods in the Algae, Some,
23
pusiola (Thelephora), 208, var. terrestris,
209
puteana (Coniophora), 197
Pyropolyporus robinicej 133
R
radiata (Thelephora), 210
Randia aculeata, 411; Gaumeri, 410;
Purpusii, 410; truncata, 411, 418;
xalapensis, 411
Ravenclii (Cyphella), 371, 372, 382
Ravenelii (Cyphella), 373
Bavenelii {Thelephora), 207
regularis (Thelephora), 20G, 207, 926
Rhipidoncma, 199
roseus (Craterellus), 332
Ruellia Parryi, 410
s
Saccardoi (Cyphella), 373
Salivary organism, The identification of
the most characteristic, and its relation
to the pollution of air, 47
Sambuci (Corticium), 191
sanguinolentum (Stcreum), 194
Scenedesmus sp., isolation of, in pure
culture, 34
Schizophyllum commune, destruction of
sap-wood of mesquite by, 248
Schramm, J. R. A contribution to our
knowledge of the relation of certain
species of grass-gi'een algse to ele-
mentary nitrogen, 157; Some pure
culture methods in the algae, 23
von Schrenk, Hermann. A trunk dis-
ease of the lilac, 253; Two trunk
diseases of the mesquite, 243
scissilis (Thelephora), 204, 226
Sclerocarpus, elongatus, 412; Schied-
eanus, 412; Schiedeanus var. elongatus^
412
Sclerotinia cinerea (Bon.) Schroter, A
study of the physiological relations of,
291
scoparia (Thelephora), 222
Sebacina, 190, 197, 198, 199
Senecio, 263; alatipes, 270; albonervius,
275; aUenus, 278; alvarezensis, 275;
angulifolius, 277, var. ingens, 276;
anisophyllus, 278; appendiculatus, 265;
arizonicus, 267; brachyanthus, 277;
Breweri, 265; callosus, 271 ; canus,
266; carnerensis, 273 ; chapalensis,
277, var. areolatua, 278; Chrismarii,
278 ; convallium, 266 ; cordo vensis,
277; Coulteri, 272, 273; diffusus, 264;
doralophyllus, 271; Douglasii, 275; ex-
imius, 271; fasiigiaius, 269; fiUcifolius,
274; Hartwegi, 280; hederaifohua, 278;
h3q)erboreaU3, 264, var. columbiensis,
264; hypomalacus, 278, 282; iodanthus,
272, 286; Kerberi, 279; kernensis, 266;
Klattii, 281; laiipes, 271; longilobus,
274; macropus, 267; multilobatus, 265;
neo-mexicanus, 265, 267, 268; oaxa-
canus, 279; oreophilus, 267; oreopolua,
268, 284, forma aphanactis, 269;
Picridis, 274, 275; prsecox, 281; prion-
opterus, 270; prolixus, 264; purpuras-
cens, 273, var. fossancrvius, 273;
reglensis, 2S0; resedifoUus, 264, var.
columbiensis, 264; roseus, 281; roseus,
281; stoechadiformis, 274; subauricu-
latus, 270, 290; teliformis, 275; velatus,
280, 288; viejensis, 271; vulgaris, 274;
Warszewiczii, 270; Wri-htii, 269
Senecionea), Descriptions of North Ameri-
can, 263
Septobasidium, 198
Silicic-acid jelly, preparation of, 39
sinuosus (Craterellus), 337
Siphonoglossa, Greggii, 409; Pilosclla,
410
Sisyrinchium alatum, 407; alalum var.
anguslissimum, 406; angustissimum,
406, 4H
Skepperia, 343, 345
432
[Vol. 1, 1914]
ANNALS OF THE AIISSOURI BOTANICAL GARDEN
Solcnia, 358, 372; anomala, 373, var.
arbicularisj 373
gpatliularius (Cratcrellus), 345
Sphaeria focxda, 106
epiculoHa (Thelephora), 225, 226
Spirogyra sctiforniis, isolation of zygo-
eporey free from bacteria in, 37
spongiosum (Stcrenm)^ 215, 216
Staining, A method for the difTerential;
of fungous and host cells, 241
Stemodia, lanaia^ 409; linearifolia, 409;
tomentosa, 409
Skviodiacra linearifolia, 409; icmcntosa,
409
Stereum, 192, 193, 198, 199; albobadium,
destruction of sap-wood of mesquite
by, 248;^ calycvlvs, 338; Leveillianum,
destruction of sap-wood of nieequitc
by, 248; sanguinolentum, latex in,
194; epadiceun], latex in, 194; i<pongio-
fum, 215, 216 ^ _ _
Stiehococcus bacillaris, isolation of, in
pure culture, 32; relation of, to elemen-
tary nitrogen, 178, 179, 181; S. subtilis,
isolation of, in pure culture, 32
Stigeoclonium tenue, isolation of, in pure
culture, 36
suhcyanea (Cyphella), 380
Bubgclatinosa (Cyphella), 370
Bubgiganteum (Corticium), 197
Bulphurea (Cyphella), 360
Syringa vulgaris, trunk disease of, 253
T
taxophilus (Cratcrellus), 339, S60
Temperature, effect of: on growth of
alga^ in pure culture, 180; on acidity
of ripening tomato fruits, 239; on
cellulose hydrolysis by Sclerotinia cin-
erea, 311
lephroleiica (Thelephora), 213
terrestris (Thelephora), 199, 218, 219,
222, 228
texensis (Cyphella), 371, SS2
Thelephora, 193, 196, 197, 198, 199;
albido-brunnea, 214, 216, 22S; angus-
tata, 205; anthocephala, 202, 203, 226;
biennis, 215, 216; caespitulane, 204,
S46; caryophyllca, 207, 209, 226;
corbiformis, 211; cornucopioides, 205,
212, 346; cuticularis, 216, 228 ; dcntosa,
224, S46; fimbriata, 222, 226; griseo-
zonata, 221, 228; hisccns, 207; inty-
bacea, 217, 220, 228; laciniala, 219
lutosa, 216, S46; magnispora, 211
226; multipartita, 205, 208, 226
odorifera, 214; palmata, 201, 2{]3y226
palviaia mnericana, 201 ; pannosa
207; perplexa, 223, 346; pusiola, 208
var. terrestris, 209; radiata, 210
Ravcnelii, 207; regularis, 206, 207
£26; scissilis, 204, 226; scoparia, 222
spiculoea, 225, 226; tephroJcvca, 213;
terrestris, 199, 218, 219, 222, 228; vialis,
213, 228
Thelephoraceac of North America, The,
I, 185; II, 327; III, 357.
Thompson, C. H., Greenman, J. M.and.
Diagnoses of flowering plants, chiefly
from the southwestern United States
and Mexico, 405
Tiliffi (CypheUa), 364, 382
Tilice (Pcziza), 364
Tomato fruits, The effect of certain con-
ditions upon the acidity of, 229
Tomato plants, transpiration of, 15
trachycha:ta (Cyphella), 379
Tramctes, 82, 138; Ahieds, 143; hfspida
142; laclea, 144; maUcola, 140, 141;,
mollis, 141; nivosus, 143; ohicnsis, 128;
Peckii, 140, 142; Pini, 142; rigida,
189, 140, 141; rohiniophUa, 104; ruhes-
ceriSy 145; sepiuni, 139, 140; serialis,
139, 141 ; suaveolens, 140
Tran8]M"ratiun, The effect of surface
films and dusts on the rate of, 1, 351
I'rcmellodendron, 197, 199
Tulasnclla, 198
u
I
Ulothrix 8p., isolation of, in pure culture,
43
Ulva Lactuca, Indications regarding the
source of combined nitrogen for, 229
unicolor (Cratcrellus) , 340, 341, 342,
348
V
vagum (Corticium), 197
Vaucheria sp., isolation of zoospores free
from bacteria in, 36
Vaughan, R. E. A method for the
differential staining of fungous and
host cells, 241
vialis (Thelephora), 213, 228
villosa (Cj'phella), 365, 382
w
Watering device, automatic, 3S6; con-
struction and use of, in transpiration
experiments, 351
X
xavthopvs (Merulius), 336
Xylaria obovata, 225
z
ZephjTanthes chrysantha, 406; Egger
siana, 406
i
Volume I
V '
) I.
i
■5
Number 1
n
tm ^.
m
Annals
of tte
Missouri Botanical
Garden
<
Introduction
Contents
March, 1914
• • •
1-11
The Effect of Surface Films and Dusts on the Rate of Tran-
spiration B. M. Duggar, and J. S. Cooley
r
Some Pure Culture Methods in the Algae. .Jacob R. Schramm 23-45
1-22
Identification of the Most Characteristic Salivary
ism
August G. Nolte
47-80
The Polyporaceae of Ohio i'. O. Overhoits 81-155,
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V
^
Annals
of the
Missouri Botanical Garden
A Quarterly Journal containing Scientific Contributions
from the Missouri Botanical Garden and the Graduate Labora-
tory and Faculty of the Henry Shaw School of Botany of
Washington University in affiliation with the Missouri Botani(*al
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George T. Moore
Editorial Committee
Jacob R. Schramm
Benjamin M, Duggar
Information
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