an: warm: STUWS are
gmm‘m MARQAMACEA' is. 5.3.

Thai: for the ham at EL 3‘
meme“ saws COLLEGE
Lidia Jean Saunders Wag“;
19505.

 
 

0-169

 

LILLIPU‘IA :thAichcmA (N.3.)

   

:l ' > , ' ‘10:" . ' ‘
'l' “‘fl . Irw|5mfi \
g P 1

k‘ g” Q. t'. . ’ . . ' . I

“M“? {It It '.

This is to certify that the 5
thesis entitled !
P

BIOLOGICAL SIUDIrli‘ OF

‘r‘r-sw

presented by I

LEILA JEALI LBAUH 332.5 WICKER

Date

has been accepted towards fulfillment .
of the requirements for

u—4

' 1:. 3. degree in B0 TAIIY

 

  

 

L:
p
i
- 1
DdCUMBUh g, 1950
|

 

-§ - _._._.._.--..A__-. -

.ah‘JJ _. -_.

hm.“ 4-. .2

o
.. ‘50—. --.-~—_- -t--._-." w..- 0-.
' .

‘. w.'-——-.._ . _
.

v A ‘1
A[{l.l[[[ll‘llltlAlll‘l‘allll‘flllllllllll‘l [Ill

BICLCGTC_L STJDIES CF LILL FTTIA IARGARITHCZAV‘(T.S.)

 

'fi‘ «’3 C, T ‘

-L nubugs
Submitted to the Graduate School ofifiichigen Stete
College of Agricultu e and Agglied Science in
pertiel fulfillment of the requirauents
for the degree of
f’L—.SZ"ZZR Ur" SCZE’CE

Degertment of Botany end Ilent Pethology

1950

THES‘Q.‘

m "T ‘--‘ "f’ C 7""?- m“
.Lgflwbfl ’5 ‘-—a— -Q

I. Introduction.................................
II. Review of Litereture.........................
III. Heteriels and GeneIul Tethods................
u. Cult:res.................................
E. Hedi; used...............................

A v

u. Litro;en “rd Cerbon EquiVelents..........

IX]. EX"eI‘i£fientgl ".'-'OI"':I...........................o

)1
C)

fl ~ ' ~‘v . V. -- V.“ "“fi . ' ' Y‘
H. EBLQflORSulg of tne L111 {431“ an“

 

 

’2 rec
Ub\~\ V0.00...OOOOOOOOOOOOOOO0.00.0.0.0...

lo Sirlf:le Cell 1801.4“..i0f28.................
:. Germinetion of Uoniaie.............

b. Germinetion of ”scosgores..........

B. Effect of Various Sectors on Growth end Reproduc

l. Tutrition.............................
u. Or: nic medi-.......................

b. 837-: 1181310 118511;.....................
1) Cerbon metebolian.................
a) ritrogen sources..................

V

iggiltoooeoeecoooeooeeoooeooooooeooeoeo

De
I?"

:3. Effect of Tel.i;~‘er.1tlll‘e end 1:3 on 31‘07-"0h And

Reproduction..........................

C. Errect of Verious Eectors on the Texonoeic Cherec-

teI‘S 3f the E‘uULUSeeooooeooooeoeeeeoooeooooeoooee

l. Yutrition, Temgereture end Iydroreh Io

n Con-

centl'u-tiOUOOOOOOOOOOOOOOOOOOOOOO0.0.00.0000...

fl

i... o:,“"t:18tic Ltd CI‘gfliiC :1‘.edivlooo0.000.000.0000

v. DiSCL‘sSSiOFLOOOIOOOOOOOOOOOOOOOOOOOOOOOOOOOIOOIOOOOOOOO

1"}! .3 ROM

Pefe

}_.J
t"

FJ
U1

(,1.‘
U]

(a
U“:

U]
0

VI. Saggrr,’ er‘d Conclusions”........................
.‘JII. .,-p:-‘.-‘evze.ixoool.o000.......OOOOOOOOOOOOOOO0.0.000...

YJ’III. Liter;xt°~~l‘e Citedeo0.00.00.00.0000.000000000000000

T-ble

Table

Table

Table

Table

Table

stle

Teble

Teble

Table

Table

Teble

II.

III.

IV.

V.

VI.

VII.

VIII.

‘1
1)..

XII.

XIII.

XIV.

“'1 ".r\

T in e“‘m1
HIV}. U .3 5-1;.L‘4Q

Amounts of eager end cerbon ecu irulerts in medi;

eIAA‘tJlOBredOO...O.O...OOOOOOOOOOO...0.000.000.0000.

Growth on verious orginic medie es re‘resented

00101137 61%;.Bt6r8000O.00......OOOOOOOOOOOOOOOOIOOOII
Growth 0- various organic media es retresented by
”screec011c observotions.........................

Ten deys' gnonth of Lilligutie g”, -ritocee et 20 0C
on virious synthetic medic es represer.ted b“ “Vere ge

 

 

diameter of 5 plates.............................
F-11ting of lilliputi1 mergiritecee on virious

 

ated by ;;;.;.croscopic

O‘Dservgtj-OIES0.00000000000...OOOOOOOOOIOOOOOOOOOOO

synthetic media as repres

iritlcea at
recite,

Bight doys' growth of Lilligvtie For“
"ion ‘”
uu

v on lectose medium with REVS

 

i hwy-r
n 11-4.00O...OOOOOOOCOOOOOOOOOOOOOOOOOOOOOOOOOOOOO

Eicht days' growth of Lilli;

o ,1ti, rrrgeritecee at
as C on gsloctose medium «its Lies

l
:rigine,

 

or e-esp1

H.

n Ell--OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO00.000.000.00.

 

Eirht days’ gromth of Lilligutie m _§critecee at
uOC on glucose ledium with Etta or e-esteregine,
W

6Kt dw s' gr0'
Siczvs

it; of Lillipitil
1.3610311 Witil :1.de

11,.
in “$.00...eooeoooooooooooeooooeooe0000000000000...

.1rgcritrce3' ct
or “-uva\J-l-f

 

LO QC 011

fie ,

n I

‘ 1 1 . °v~“ Q. n i.’\"_ Q
01 sillliitle #11
or J." As;

Eight d1 grouth
9_On

o o on meltose median witthKDB

riritace; at
'"1ne,

 

in MELOOOOOOOOO0.00.00.000.00000000000000000COOCO:.

Ten days‘ growth of Lilligutie 'lrgiriteces at
£000 st Verious ligit on dirfeient

£36—i1LOOOOOOOOOOOOOOOOOO00......OOOOOOOOOOOOOOIOOOC

 

'..1_,\ ,.:.:..' r.
In ton alblED,

The effect of various light intensities on the fruit-
in- of Lilligutiu mergiritlcei in various medic as

\J

epresented by microscogic ObSerVutiOnS..........o

 

Effect of light, inte..1tte t light: u.d dorkness on

CCflidi-—ll Sizes.00.00000000000000500000000900000...

Effects of hydrogen ion concentrction, tem2er tare
etc gedie on the growth of Lilliguti: 11:; rit ce :1..

 

14

I7

13

(Fl
{"1

1

CH

Tuble

Toble

Teble

Tgble

Table

Table

Table

Table

XV.

XII.

XVII.

XVIII.

1r-“f’
ILdeo-

“f..-

.‘
-Hs.

XII.

‘1"!r'

14L]: 0

-l'v-f‘or-

.u..‘..L.1_

1': JV- 0

amp-1r
INA-5 O

XXVI.

“4|“

siiects oi‘ h"cr05' en ion conce: r1.'or, temier1t1re
end gedio on fruiting end growth oi‘ Lilligutie

 

F'Y‘T" ‘n':‘tacp'
_.i_.L»-~ .LJ. 1- u-0000000000000000.000000000000000000000

 

Uptimun temgerotures for growth of Lilligdtie
m rgLrit ceo on VMIIOUS 171rs gt v ried hydrogen ion

 

 

concor'itl‘ufiiOI’lSoooo0000000000000000000000000000.000

 

 

 

ETfects of erv1rorne t 1 conditions on the fro th of
Lilli;utii 71;; Joe: es re 1ooc ted by hieroscooic
1.';€AS.J.I‘3.GI$S. ”Daub ug'wr 1110 0000000000000000000000
Effects of environrentol con 1di+ one on the growth of
hilliiv tie er~.:"ituceu 13 1e; rese ted by microscoric
..1L.;.~Jx-.lu.ETLL150 -LICEA‘O sci-KAT 141-5.COSOOOOOOOOOOOOOOOOOOOC
Effects of environment I c:rditiors on the frthh of
Lilligutid zlrfiritcced uS regrc n ed by microsc031ic
171:3-.~S_'CC‘8.1.€‘?:2tS. -C-.t p.193: ;:: 70C-e 0000000000000000000

environgertel cohditions on tge gro ;h of
C‘

 

~ ‘ .‘- ‘. n .I-"_ . 1 —-‘ - g. A‘ v a A

Ll'lltxtl— 15 11th ed es e3resented 3 micrODco ic
4 'I - ’1.J- ‘ r J" —- 'v "‘TT I

;1-6‘.1buI‘&:-.1czT1oS. HIE-gt: 1.5-1.4: I"; 70400000000000.00000000

 

Effiects of environsertel ccrditions on the grouth of
Lilliyutii xflrL_1it;cee es regresented by mi roscobic
:18 18:11‘831811‘58 0 .iE Qt “For PI: 3.0 0 0 o o o o 0 0 0 o 0 0 o 0 0 o 0 0 0 0
Effects of environmentel conditions on the {To th of
'Lilli*utio m rg1rit1ce; us represented by microsc03ic

 

A
2",". ‘1 - ‘. ‘. - h I ' I ‘ 17'}, T 'I 'p l a - '-~ ' w—T " A
44A: 1...) fire“). ".1. tb . 4: 1‘15 5) '.uQC..L .L l 80- 1.15:1]? {WI 0 ob o o 0 o 0 o s o o o 0

Effects of environmentil corditions on tie fire th of
Iilli3utid nnrgsritoceo 1s regreserted by microscotic
fileaS‘JreiJentS. :J‘I'ies IilOCIiIi ed Q. _-...r PI“: 6050 o 0 0 0 0 0 0 0 o o
‘fects of environmentsl conditions on the éronth of

E1
Lilligutis he::.11it-ces es represented by microscopic

 

:easureuants. iries modified dyer pH 7.0..........

Effects of environmentol ccmditions on the growth of
Lillll‘t’ morgirit-ceo gs reoresehted by microscopic

 

1113115 Lire .CITLS. Tie 3 11100111in Cé :LI‘ P‘Il 70 4000000000.

hi‘i’ec ts of environ ehu .1 conditions on t e growth of
Lilliputia mhrgsrithcee cs reyresented b3 microscopic

 

LBCS.LreuLeE'LtSQ J-I'ies llfidified as)? 1:1: 3.0..........

(,1
0:

(<1
U 1

C 13
{J

40

41

4s

W
01

1b
0)

1—‘lult e I O

E gure

Eiglre

Figure

Plate III.
Figv-‘Lre

Figure

Flat 8 n7 0
Figare

‘7“. '
21?.er

'1". 4- *
lebe ‘JT.
rlgmre

‘1' H
.L'lgrl‘e

Fiyure

l.

O
H.

1.

gm 4‘ 7‘1 31' r" f:-:‘1’1
LI’JJ- LI) .1. s;— ‘J..L 1.40

Gerglimtior; of a con.idi;u;. . . .

Germinqtiou of at escospore.. ..
conifidl C113:€4'J?;;-tus.oooooooooo co
COnidiol alJiJvLI'CLtLLSOOOOQOOOOO o .0
Conidioghores aid Cenidiel Leeds.

meature
“scosgores shcuing

H.

Chi Cali; A

Terious forzs of conidic.....

conidiul ILeHLdSOOOOOOOOOOOOOOO

1"eriti‘leCiLLooooooocoo0.0.0.0..
future ascospdres............

1.:QtUI‘e {1803.SooocooooooOoooooo

CI
01

(n
0“

(n
C)

.‘l
"5
g.
L“;

n. .
urchi'.

Grcph

5.

"a“! . .\ m T',"‘
v.3. U2 BRA-I‘D

The effect of Veryinr gacntities of sugcrs ugcn
the grawth of Lilliiutiq ficTZ§TIthBQ using

potissium nitrcte as the nitrcgcn scirce.......

 

The effect of verying Quentities of sugcrs ugon
the growth of Lilli;utie Jir:;rit; c; using

n
‘ V
v - n i W, . - v - . . 4' . —q - I \
e-ccpeiegine ca tce nlurccen soirce............

 

-r

The effect of various media on tge growth of L.
t

. ,‘,1' 1‘. . .f“l'
fwliz-ll 9069. In ll‘:_Altoooo000.000.000.00000000000

 

 

Tne eifect of Various ¢ed
.1-

ii on the growth of I.
1L1:}Tit;ceJ in in cruittent liyhto-------°--°°°

 

The effect of virious media on he grcxth of _.

i.L.:f';'_jx:r]."i+Lice-.4. in CLI’kIEeS-S o o o o o o o o o o o o o o oo o o o o o o o o

 

L3

hfiflIXTVhfirfiimS

I wish to eigress my upgrecihtion to Er. C. J. IleXOpoulos for
his Vuluuble time, ugprOVel oi‘the thesis problem, the photosruphs
used in ELLTES IV and V, for his guidcnce, yetience, and criticisms;
Dr. Ernst Bessey for the original cultures, suggestion of the problem
end miny’trehslations; Ir. J. H. Seumun of the Depsrtuert of Eoreign
Lunguyge for the trunsletion of the techniCul description into Latin;
firs. Isrie.Loour for muking sVuileble muny meterisls necesssry to
complete this wo i; and members of the fuculty end stuff of the
Botuny end Bacteriology Degurtnents whose coOperution mhde this thesis
possible.

I esgecielly went to theflk Dr. Rey Yelson for instruction in
photosrsphic work, use of his ecuigment, end for Valuable yeurs of
truining in lsborutory methods.

L. I. S. III.

I. IXTRCDUCEION
A fungus, growing on stable manure Was collected by Dr. Ernst Bessey,
at East Lansing, in the full of 1944. Upon examination, geurl-like
structures and smull glistening hyaline mucous droplets were found on
the substrate.
In order to determine the taxonomic position of the fungus, it
was necessary to isolate and germinate single cells, and to study the
behavior of this fungus under different conditions of nutrition, and
environmentul factors. These studies constitute the body of this thesis.
The germination of a single cell (conidium or escospore) resulted
in the production of the asexual stage belonging to the form genus

Gliocledium, and the perfect stage which proved to be a species of

 

Lilliputia.

 

As will be seen from the body of this thesis, the results of the
present study indicate that the fungus under consideration is e new

Species. it the suggestion of Dr. Bessey the name Lilliputie mergnritecee

 

is hereby proposed for this fungus in accordance with the following
description:

Lilliputig mergnritacea, sp. nov.

 

Kycelium hyelinum, laeve. Conidiophoruel‘- ramosee, symmetricee,
plerumque 411 x 29p (280-481 x 9.4-2Ip), 2 - 4 ramis 4.3 - 5.5 x 25.8 -
54.4P, singulis hubentibus 3 - 4 metulus 3.4 - 4.5 x 21.5p, qune singulae
2 phialides 2.2 - 5.4 x 25.8 x 45? ferunt. Trie genera conidiorum
hyalinorum, luevium, unicellulurium generantur: Conidiu ellipsoidee 1.6 -
2.4 x 4.4 - qu, OVelia 5.4 - 5.8 x 6.7 - 7.5p et spheericu 6 - 2p ordine

in phiulibus generate et gutte mucosa cohuerentia. Perithecie 400 - VOQP,

L
plerumque 509p, per spire: mycelihgenerete; cerosu, capillis pcpillutis;

l H . ‘ l l ‘
l' 9 V I. “I /‘.‘.‘ I .1' I UM"?
\ f .s. \u , 7 ,
KIM 5 I

I
I,

pariete eorum laxo et duio prime, concrete at 510111 in .rsuturitete. hsci
Qqfi-3ox2L.*- w,md,.6437g (Asumvtayn*¢ouat 'Q/

hyulini’ spines hebentibusrquee lonyrtudinew=mode—diametri spores in
asco sedentis hebent. lscoigorue Ineturue 19. 3 - 21. .5» spinosue, ferruzineo-
brunnene. Spinue 1.9 - 2.8g longue.

Ab E. A. Bessey collects in East Lansing, Hichigun autumno enni 1944
ex stereo.
English description)

chelium hynline, smooth. ConidiOphores spuringly branched, symmetricul,
average 411 x 20p with a range 61‘ 230 - 431 x 9.4 - 2111, two to four
branches 4.3 - 5.5 x 25.8 - 34.4? each having three to four metulae 3.4 -
4.3 x 21.5p which in turn beer two phiulides each 2.2 - 3.4 x 25.8 - 43p.
Three types of hyuline, smooth, unicellular conidia are produced; ellip-
soidal 1.6 - 2.4 x 4.4 - lQP, ovoid 3.4 - 3.8 x 6.7 - 7.8P and sphericul
6 - 9P, conidia produced successionally upon phiulides and held tOgether
by a mucilaginous droplet. Perithecia 400 - 709p, mostly 5OQP, produced
by the coiling of mycelium; waxy, with papillete hairs; Well loose and
hard at first, compact and soft at muturity. “sci hyuline 24.4 - 30 x
26.5 - 55.9P, subglobose, bulging with 8 eculeate hyeline escospores
having spines % diameter of the spore in length while still in the ascus.
Mature ascospores 19.3 - 21.5».echinulete and rust brown in color. Spines
1.9 - 2.8p long.

Collected by E. e. Bessey in East Lansing in the fell of 1944 on

stable manure.

II. REVIEW OF LITE'AIURE

Lilliputia Boudier et Patouillard (5) is described as somewhat

 

resembling Terfezia in so far as the spores are concerned. The perithecia
are described as having a cortical portion which is very thick and much
differentiated around the gleba. A great number of asci are found in

the jelly-like matrix. The spores are yellow-brown inside a very white
perithecium.

In 1887 Winter suggested a possible relationship between Gliocladium

 

penicillioides Corda (3) and Eurotium insigpe (26). He mentioned conidia

 

 

but gave no characteristics for them. Later Fetch concluded that this

was not the imperfect stage for E. insigge but named _G_. macrgodinum in

 

its place.

In 1910 Bainier (2) described Gliocladium prolificum. a perfect

 

stage was feund which was thought to belong to the Perisporaceae.

In 1946 Dennis and Wakefield (9) suggested that Eurotium insigne

 

 

 

Winter, Lilliputia gaillardi Boud. et Pat., and g, penicillioides Corda,

are the sane species and the new combination Lilliputia insignia was

 

proposed for this fungus and the other three names are synonymous.
This was placed into Eurotiaceae near Eurotium.
Eurotiun was first suggested as the perfect stage of the genus

Gliocl:~<‘iiuIII. In 1946 Dennis and Z'Jakefield named Lilliputie as the perfect

 

stage. The above includes a review of those species of Glioclldimn hav-
ing a sexual stage reseibling that of our fungus, and those associated

with the two species of Lilliputia hitherto described.

 

III. IILITEPZLVLS [JD GEEK-J, LETHODS
A. Cultures
0f the twelve original cultures made by Dr. Bessey in 1944, only

one, #11, was revived and used for further study. Others were either
dead or contaminated. manure extract was used in an attempt to revive
these cultures. The medium.Was prepmred in accordance with the following
directions:

Fill one half of flask with horse manure.

Add water to the top of flask.

Steam and stir frequently'for one hour in an Arnold Steamer.

Filter through several thicknesses of cheese cloth.

Sterilize at 15 pounds pressure for 1% hours.
This extract can be stored for further use.

Eff: Manure Extract Agar

 

2cc. Manure extract
2 gms. agar
9800. distilled water
Cultures were examined at intervals in.mounts of lacto-phenol, or
lacto phenol and cotton blue which stained the fungus light blue showing
the phialides well.
All measurements were made in Lacto phenol mounts with a Filar
Microme ter .
To prevent contamination Petri plates containing fungus cultures
which were kept in incubators for 10 days, were sealed with scotch tape.
Previous experimentation showed no effects on growth due to a possible

decrease in oxygen.

III B. Media used

 

Hedia used throughout this work are listed as follows:

Coons synthetic agar
7.2 gas. saccharose
5.6 " dextrose
1.25 " 1:35:80
2.72 " EELI‘
2.02 .. mg
1.5% agar
1000 cc. distilled water

 

4

Westergaard and Kitchell's synthetic medium
.1. £111. I305
.1 an. KHnPO
t“ .c 4:
.05 " ihg804
.Ol " CaC13
.01 " NaCl
5ug per liter biotin
2 gm. sucrose
1.5% agar
.lcc. Beadles'trace elements (pg. 15)
100 cc. distilled water. Adjust to pH 6.0

 

Richard's synthetic medium
1.0 gm. KKOB
.5 " KHPO4
.25 " ligSO4
Trace FeC13

5.43 gms. baccharose
100 cc. distilled water

Fries synthetic (less tartrate) Fitchell's modification of Friesl

 

so as to remove the tartrate.
4.0 gas. Anionium nitrate
4.0 {2315. KHBPO4

.2 " N11Cl

.2 " Ca012
1.0 " KgSO4.7H30
1000cc. distilled Water
adjust to Ph 6.0

 

 

lOriginal Fries formula (double strength) (3) as follows:
2.0 gms. Ammonium nitrate
10.0 fins. Ammonium tartrate
2.0 gyms. ‘12}?04
.2 Ems. NaCl
.2 gas. CaClg (anhydrous)
1.0 {21:15. 153504.7320
1 liter water
adjust to pH 6.0
(Sucrose, biotin and trace elements not added)

Czapek's synthetic ajar
.5 gm. Kg504
1.0 " KEEPO4
.5 " K01
2 . O " 133503
3.4 " saccharose
lOOOcc. distilled water
1.5% agar

 

Corn meal beer agar
100cc. beer
20 girls. Difco corn meal agar
2 " sucrose
water to lOOOcc.

 

Wheat agar
100 gms. wheat
lOOOcc. distilled water
1.5% agar
Infuse meet for one hour at 500C. V(wrap in cheese cloth and
suspend in water).
Decant, add agar, steam for 20 minutes.
When clarity is necessary, filter through glass wool.
Sterilize at 12 lbs. pressure for 20 minutes.

 

Bacto beef heart infusion
This is a liquid infusion medium.ibr the cultivation of
fastidious organisms.

Oat afar
100 gms. ground oats
lOOOcc. distilled water
Inflision for one hour as in wheat agar
1.5% agar

Potato dextrose agar
200 gms. peeled and sliced potatoes
lOOOcc. distilled water
Steam.for one hour, decant.
1.5% agar
15 gns. dextrose

 

Bacto bean agar

22.5 gms. Bacto bean agar

lOOOcc. distilled water

Sterilize at 15 lbs. pressure for 20 minutes.

 

Rice agar
100 gms. polished rice
lOOOcc. distilled water
Infuse for one hour as in wheat agar
1.5% agar

Red clover st ems
Four 2 inch long stems placed in test tubes.
5cc. water (distilled) in each tube.
Steam sterilize (one hour each day for 3 successive days.)

 

Carrotgylugs
Cut carrots to fit test tubes in slant-like strips.
Add few cc. distilled Water
Steam for one hour for 3 successive days.

 

III, C. Eitrogen and Carbon Eguivalents

 

The basic medium (Coons synthetic agar) was sugar free.
Two constant sources of nitrogen used were potassium nitrate and

a-esparegine. The 2.029 grams of KYO used per liter of Coons synthetic

3
medium contain .280 gms. of nitrogen. Asperagine having the same equivalent
was used (1.320 grams of a-esparagine contain .280 grams of nitrogen per
liter.)

With these two constant nitrogen sources, various molurities of
sugars were used. Two sets of each molarity of sugar were prepared in

500c. aliquots and each set run in triplicate. KH03 was used as a

nitrogen source in one set and a-asparagine in the other (TABLE 1).

7‘ V‘ ’V‘!

-

n-rvfi

 

 

 

 

 

 

 

TIE 31. “ICU?“S ,1 31312 ;”D C ”301 “ JLTI..L;:13 IE; :LDIA ELELOYZD;
ITI'I‘.‘ CC T42; COLSTA. T.
Eiglur ' 0.1.5. ye: 11.15}; ' Gas. 961" Gas. C
Sugpr Conc. Flgsk Yo. [50 m1. aliguot) liter per liter
.03 1 .297 5.95 2.15
Dex- .06 2 .594 11.39 .33
trose .09 3 .392 17.84 6.43
v.1 .12 4 1.139 25.79 3.63
193.17 .15 5 1.436 29.74 10.79
.03 1 .510 10.27 4.32
Sacchar- .06 2 1.030 20.53 3.64
058 .09 3 1.540 30.30 12.96
1'.'.?. .12 4 2.050 41.06 17.27
342.2 .15 5 2.560 51.32 21.59
Lactose .03 1 .510 10.27 4.32
r. '3. .06 2 1.030 20.53 3.64
34 2.2 .09 ' 3 1.540 ' 30.30 12.96
.12 4 2.050 41.06 17.27
.15 5 2.560 51.32 21.59
Galac- .03 1 .270 5.40 2.15
tose .06 2 .540 10.30 4.3
H. E. .09 3 .310 16.21 6.43
130.15 .12 4 1.030 21.61 3.63
.15 5 1.350 27.01 10.79
331- .03 l .510 10.27 4.3 2
tose .06 2 1.030 20.53 3.64
M. W. .09 3 1.540 30.80 12.96
342.2 . 2 4 2.050 41.06 17.27
.15 5 2.560 51.32 21.59
1. . .
Lolecular nelgnt

IV. EIG’EEII 331‘? TH. T? \FK

A. Relationship of the Lilliputia and Gliocladium Stages

 

1. Single cell isolations

a. Gennination of conidia

 

A duplicate series of water’hlanks ccntaining various amounts of
water, at.1cc. intervals from..1cc to lcc, was prepared. One pair of
water blanks for each interval was inoculated with conidia obtained by
applying a sterile platinum.wire to a single conidial lead. Each tube
was shaken to separate the individual spores so that when the contents
was poured onto a plate containing solidified 2%.manure agar and the
plate rotated, the spores spread fairly evenly on the surface. Each
plate of filtered 2% manure agar contained about 150c. of agar. The
plates were inverted on the stage of the microscope and observed through
the low power objective. 4s spores were found, an India ink mark was
made on the glass so that the spores could be located thereafter. Best
results were obtained with.the .500 water blanks. The spores were spread
out sufficiently so that single germinating spores could be cut out
readily and transferred to 2% agar plates. The majority of conidia
had germinated 13 hours after inoculation. Ihrphal tips were cut from
the resulting colonies and transferred to test tubes containing 2% .
manure agar. All single cell isolations and hyphal tip inoculations

produced the Gliocladium stage within a few days, and the perfect

 

stage within 10 days (Plate 1, Figure 1).

Iv. A, I, b. Germination of ascospores

 

The same method used in genuinating conidia was also used in
determining the best dilution for ascospore isolations. In this case,

however, a single mature perithecium (the surface of which sometimes

bore conidia) Was transferred to each sterile water blank and crushed
with a sterile glass rod. Each dilution was shaken for 60 seconds to
separate the ascospores, poured on a cooled 2% filteredl manure'agar
plate. The plates were rotated, and after the excess moisture had
evaporated, they were inverted on the stage on the micros00pe and marked
with India ink to indicate positions of the ascospores. Best results
were obtained with one crushed perithecium in .5cc. of sterile tap water.
These plates were examined daily until the agar had almost dehydrated but
no germinated spores were found. This procedure was repeated three times
with the sane results. '

Heat was used in an attempt to germinate ascospores. four sterile
test tubes containing ascospores suspended in .5cc. sterile distilled
water were prepared, and placed in water baths at the following temperatures
indicated below, and poured onto 15cc. 2Q manure agar.

Tube fil-- 7000 for 10 minutes.

Tube #8-- 90 C for 5 minutes.

/

Tube ga--eooc for 10 minutes.
Tube 34-- 5000 for 10 minutes.

30 germinations had occurred up to the 10th day.

Hydrochloric acid was chosen in a third attempt to germinate the
ascospores. Since the fungus had been found on manure, the following
experiment was set up. According to Dukes (10), the H01 range of the
stomach of the horse lies between .14 and.21fl. The statement that the
stomach emptied its contents conpletely within two and one half days was
taken into consideration. The following dilutions of 301 were prepared:

.l4%, .153, .163, .175, .13fi, .192, .20%, and .21fl. Two tenths of a mi.

1For all single spore isolations, media were filtered through washed
glass wool.

10

of each dilution was transferred to a test tube, sterilized, and inoculated
with one mature perithecium which later was crushed with sterih rod. Three

sets of dilutions were nrenared' one set "digested" one da' another tn
1 .. 3 - 3

d

0

days and the last, two and one half days.

Out of five repetitions, germinations occurred only twice at .16
and 21% (PLATE 1, Figure 2). Since HCl treatxent killed all conidia
which were transferred to the water blanks with the perithecia, the
resulting germinating ascospores were known to be "pure". For this
reason all single cell germinations derived from.HCl treatment were
used or stock cultures throughout all experiments.

ascospores were induced to germinate by freezing in a refrigerator
freezing canpartment at -60C for 18 hours. A perithecium was crushed
in a sterile test tube containing .5cc. of sterile tap water and placed
in the -6°C freezing compartnent. After 18 hours, the tube was allowed
to thaw and the contents then poured onto a cooled 2% manure agar plate

and rotated. Germination of ascospores and conidia occurred.

Colonies from.all single cell isolations (ascospore or conidium)

produced both perfect and imperfect stages.

11

\

IV, B. Bil ect of Vu:ious ructors on Growth and Reproduction
l. lutrition
a. Crg.nic media
In un act.“ t to produce both stages of the ftnsus in abundur ce
and in a campurutively short time, the following nc’ia were inoculated:
out, manure (corcen tr ' ted), Bfi manure, rice, wheat, pet to de: :trose,

corn meal, beer and 23 a5urs. Liguid wheat medium and clever atens

h)”

6"

erilized by tyndullization were also used. Triplicate plates were
made of etc; nedium and inoculated with a perithecitn. Grouth measure-
ments were made each day for 10 days and all :lates mere kept at 3600.

The resulting groath :eesurements and microscopic obserVatiors are

Generally organic media produced culparatively thick yrowth as well
as both states of the fungus in a short time.

In wheat aHar, conidia and peritheciu were found in abundance
(about BOO peritrecia per plate).

Rice nedium.prod uced as mu: y perithecia but the con idiul stage was
not as ubur.d..r1to

The only explanation the author can give for the absence of growth
in concentr ated m_r.uI e a5 ur is that a new batch of manire extract had
been prepared since tie initial experiments and the manure shy'have been

too old and toxic.

'12.

.mopwan owns» no Hmpmswflc qwoe.onp pammonnoh monzwfih*.

 

 

 

 

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OH 0. m . s. m m w a m
when 232

 

13

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TLBLE III.

Growth on Various Organic Kedia as Represented by

Macroscopic Observations. (10 days' growth at 26°C).

 

 

 

 

 

Perithecial Conidial
media _production production Lwtelium
Rice agar ’Abundant; 7 'Abundant Dense growth
days after inoc.
Oat agar Very few; 7 None Dense growth;
days after inoc. atypical
Wheat agar Abundant; 5 Abundant; 4

Manure agar

'days after inoc.

'days after inoc.

Very5dense

 

 

 

 

 

 

5Lconc) None None None
manure agar Sparse; 8 Sparse; 5 Sparse
5% days after inoc. days after inoc.
Potato Dex- Abundant; 6
trose agar None days after inoc. Very dense
Water
Zfiéagar None None None
Corn-meal
Beer agar None None Dense
Clover stars None None None
Liquid wheat Pellicle over
medium None None 3/4 surface of

14

medium

IV, B, 1, b.

Synthetic Kedia

 

Growth of Lilliputia margaritacea on Coons, Czapek's, Fries,

Richard's and Westergaard and Kitchell's synthetic media is shown in

TABLES IV and V.

Fries modified synthetic medium was prepared and Beadles trace

elements were added in the amount of lml. per liter.

Beadles Trace elements

BO- .01 mg. per 1
CO“ .1 N H
E‘e_ .2 I! 1!
Mn- .02 "

Ko- .02
Zn-2.0

33
3:

Fries modified synthetic medium (see Katerials and Kethods) was

prepared in double strength (Fries 2K) and the following combinations

were made:

(a) 250cc. Fries 2X
20 gms. sucrose

3300}: 501133101?)

(B) 50cc. (a)
2 Ems. malt extract
.5 ems. yeast extract

Enough Water to make 100cc.

(0 ) 50cc. (A)
.5 gm. yeast extract

Enough water to make 1000c.

(D) 50cc. (A)

.400. biotin (prepared as 53'per co. in 50} alcohol.)

40c. vitamin mixture (see below)
Enough Water to make 100cc.

Vitamin mixture

 

B1 25 mgs.
33 12.5 "
B6 1205 "

Iantothenic acid 200 "
Para amino
benzoic acid 12.5 "

Nicotinanide 50 "

15

Inositol 100.0 mgs.
Pimelic 100.0 "
Choline 25 "
IZyd. yeast (3.1“) 250 ..
Folic acid .05 "
Water 2500c.

(E) 50 cc. (a)
1 gm. succinic acid
.4cc biotin
4cc. vitamin mixture
Enough water to make 10000.
(F) 50cc. (a)
Enough water to make 100cc.
(corTROL)

(G) 500C. wheat agar
(cormncL)

All media were adjusted to pH 6.0

Two l5cc. aliquots of each of the above media (B, C, D, E, F, and
G) were prepared, 15cc. agar added, and the sterilized media inoculated
with a perithecium.

medium 8- Growth; sparse, no fruiting

Hedium C- Growth; abundant. Perithecia and conidia abundant.

Medium D- Growth; trace, no fruiting

Isiedium E- No growth

Medium F~ fio growth

Medium G- Growth; sparse as compared to C. Fruiting; fair amount.

Westergaard and Mitchell (33) describe a possible synthetic medium

for abundant formation of the sexual stage of Neurospora.

 

100cc. of this medium were prepared. To one half of this quantity
.5% yeast extract was added, and the rest was used as a control contain-
ing no supplement.

The resulting growth was compared to that in Fries modified synthetic
medium (plus .5? yeast extract). Westergaard and Mitchell's synthetic medium
plus .Sfi yeast extract allowed the production of’more perithecia than did
Fries, but maturation of the perithecia was prolonged. Growth in
Westergaard and Kitchell's medium without supplement was the same as that

in Fries modified synthetic without supplements.

TABLE IV. Ten Days Growth of Lilliputia margaritaceu at 26
Various Synthetic fiedia as Represented by sveruge

Diareter of 5 Plates in mm.

 

o
C on

 

 

 

 

 

 

 

 

Iedia Days '

2 3 ' 4 5 6 7 3 ' 9 10
COODS iLgCir 2 206 206 306 400 4:05 500 60 7.8

1 Q '
Czapek's agar 8 15.5 15 25 26 51 55 44 47
Richard's
agar l 1.5 2 ".5 4.0 5.5 7 7 7

‘Fries Kod-
ified plus
.5% yeast .
extract 5 10 13 22 50 35 41 43 55
Iestergaard
and Kitchell's
ngar 2.5 5 10 l6 20 22 24 26 28
a:
411 measuranents recorded in mm., representing mean diameter of three plates.

17

 

 

 

 

 

 

 

 

TnBLE V. ‘Fruiting of Lilliputia margpritacea on Various Synthetic
media as Represented by Macroscopic Observations (10
days’ growth at 26°C).

Perithecial Conidial

Media Aproduction production mycelium

Coons agar ' none none ' sparse

Czapek's

agar none abundant thin
Richard's ' '
agar none none dense

Fries mod. Abundant; 4 Abundant; 6 days dense

asar plus days after inoc. after inoc.

.5% yeast

extract

Westergaard & Abundant; 7

Kitchell's days after inoc. sparse dense

agar

 

1v, :5, 1, b, 1).

 

L hou111cL.1on of Coors sznthetic :1edium see used as
study of carbon metabolism of this fun"u s. Verious gClin
.09, .12, .15 molur) of even of s=gurs .cre us d as c.
in the various GKLBriXBEtS conducted. These were comb1-ed

 

 

. w ,1 n- 5_
a base ior tne

ties (003, oUd,
1rbon sources

nith an organic

and in or; nic source of nitrogen (a-usp re ice and YTOa). The amount of
F703 and a-asgsrugine used, contuined .230 gns. of nitrogen per liter.
.gll plites were run in triplicate and each glute nus inoculeted
with a perithecium. ’.-easu exents of colony yrouth were made each day
for 10 days.
The results of these experiments are summsrized in TLLLLS VI to X
Lind h‘HuJi :) l and 2.
IV, E, l, b, 3) Yitrogen sources
1 modified Coons synthetic medium nus used es a base for the study
of nitroeen me aboli an of Iilligutia mix; ritecoa. The follet n; amounts
of urea were used nitroren sources in these exger'nents; .01, .l, .3,
and .6 ans. yer liter. Plates were ‘un in triglicete, inoculated, and
incubated for 15 dsys.
Kultose 3.03 gas. per 100cc. Results:
Q;SO4 .125 ” " " .60 gas. urea ger
Ureu .01 to .e ems. yer lOOOcc. liter troduced
KE @3104 .272 gns. per 100 cc. largest rrowth
1:: rgur 1.5-,0 (72.11151) ; jceliunl
Distilled water 100cc. searse.

C‘

OK.)

Mm onium acid Eho piste

 

M1
lile

following medium Wis prep red in tri; lieute,

I e w ' ‘ . d r r 1‘0
a s1nfle per1tnec1tm.ana incubated Ior l6 days at do C.

inoculated with

GROWTH

can" I-THE EFFECTOOF VARYING QUANTITIES or SUGARS
UPON THE GROWTH OF LILLIPUTIA MARGARITAOEA

usme POTASSIUM NITRATE As THE NITROGEN SOURCE.
NEAN DIAMETER or OOLONIEs AFTER EIGHT DAYS

AT 26’0.

 

 

.03 . 06 -09 ,l2 .l5

OWGENTRATIW W SUGAR MG”

GRAPH 2" THE EFFECT OF VARYING QUANTITIES OF SUGARS

UPON THE GROWTH OF LILLIPUTIA MARQARITAQEA

USING A'ASPARAOINE AS THE NITROGEN SOURCE. MEAN
DIAMETER OF OOLONIES AFTER EIOHT DAYS AT ZO‘O.

25 MM

 

GROWTH

 

'03 ~06 -09 .I2 .l5

OOI‘CENTRATION OF SUGAR MOL.

 

 

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Naltose 3.03 gms per 100cc. Results:

KgSO . .123 " " " .lEEgms. of (N34)2HPO4
(NH472HPO4 .122 " " " produced 16mm. of
KHZPO4 .272 " " " growth; mycelium

Agar 1.5% sparse, conidia and
Distilled water 100cc. _ perithecia absent.
Trace Fe

inmnonium nitrate

 

The following medium was prepared in triplicate, inoculated with a

perithecium and incubated at 2600 for 13 days.

Maltese 3.08 gms. per lOOcc.

£193,304: . 123 n n n

NH4N03 .03 " " " =.230 gns. N per liter
“£31304 0 2 72 II n n

Agar l O 57’;

Water 100cc. Results:

Trace Fe No growth.

IV, B. lfligfli

A plate of‘oat agar containing the fungus which had grown the
diameter of the plate, was left in a dark cupboard for a few days. When
renoved, perithecia were found in great abundance. This Was the first
indication that light was not necessary for fruiting.

In order to keep all plates at approximately the same temperature,
a large petri dish container was packed with cotton leaving enough room
up next to the container cover for 6 Petri plates. Those plates exposed
to light were placed on topIof the Petri dish container 2 feet under a
60 watt leap for 10 days. Those exposed to darkness were left inside
the Petri dish container and those exposed to intermittent 1:5:th were
placed on top of the container cover for 12 hours and inside the container
for 12 hours alternately. The container was opened only when placing
the intermittent light plates from light in darkness and in light again.
This was done inta darkroom.

The agar used, together with growth, reproduction results, and

9.5

Days' Gro

Tl3LE XI. Ten

.7 L.
“It 1;

of i~illipu’cia m rearitgcea

 

ut 2600 at Various

Light Inte.sities, on “iffeient media, as Represented by
“verses Liameters of 5 Plates inznn.

 

Tedia
Wheat a:pr r: 7.2
Rice agarng

Kcnure (Sfll agar 3H 6.9

' Liget
:35

”I

Intermittent
' L i 71." t

' T‘.‘ 1- .
J.)\_Arl“. AF: 5 S

 

Cut a:;r pH 7.0 34 66 9
Potato dextrose aga
p: 5.! 34 52 85

 

aihole

.’

lO

[‘0
(II

.:€CLS‘.11‘FiI€ntS

tubes.

 

Czapek's synthetic
medium pH 6.8

O

51

51

 

O‘o

T3313 XII.

m1
.
‘. Lit]

 

'“ Effect of Various Lifht Intensities on the Fruiting
of Lilliputia margaritacea in

Various Ledia as Represented
by Iacroscopic Lbservatio;s (10 days' great: at 25 c

-\

U“)

 

Intermittent

 

 

 

 

 

 

 

 

 

 

 

chia ' Li 7t ' Light nerh"ess
Iheat Ierithecia a 7;:ithccia; : :e i- ecia a Vanilla;
a'wr conidia forged Gags ;fter inec. E d,;s after itoc.
5 d-.:,'s after Coz'idia; .3 6:...33
irocnlgtion ' aftar 1 oc.
Rice - Eerithecia; 9 Perithecia; 3 Perithecia; 5 days
leer days after inoc. d33s after inoc. after inoc.
’ Conidia; 5 days Conidia; 8 day Conidia; absent
after inoc. after inoc.
3;“; Peritheciu a; l—‘erithecia & I’erithecia at conidia
tenure conidia conidia sparse sparse.
Agar absent
Oat Perithecia; lO Perithecia; 3 Perithecia and
lgar davs after inoc. days after inoc. conidia; 5 days
after inoc.
Potato Perithecia & Conidia; 5 days Conidia; 5 days
dextrose conidia absent after inoc. after inoc.
agar
Sterile Perithecia; lO Eeiithecia; 7 Perithecia; 7 days
whole days after inoc. days after inoc. after inoc. Conidia;
manure Conidia; 5 days conidia; 5 days 5 days after inoc.
after inoc. after inoc. -;,
Carrot Perithecia a Perithecia a Perithecia a conidia
plugs conidia absent conidia absent absent
Bean Perithecia &t Conidia; 10 days Conidia; 9 days
pod conidia after inoc. after inoc.
agar absent
Red
stem
Czapek's Conidia; 7 days Conidia; 5 days Perithecia a
synthetic after inoc. after inoc. conidia; absent*
me dium
*

If perithecia and/or conidia were not found witFin a ten day period, the
author dies not mean to state that they would not have been forged at

all 0

up in 15-20
which would

0n the contrary, in many cases both stages of the fungus did show
days, but the purpose of this section was to find a medium
produce both stages as early as possible.

GROWTH

45
4o
35
so
25
20
no

IO

lemonade

 

 

DAYS

OHAPH3 ' THE EFFECT OF VARIOUS MEDIA ON

THE GROWTH OF LILLIPUTIA HAROARITAEEA IN

LIGHT.

GROWTH

70 NM

 

 

0/
so /
O
5 5
O
50 OZAPER 8
45
4O
36 VII-EAT
RICE
30
BEAN
25
20
I 5
I O
5 v ~1I6HARO‘8
O 7 8 9 l0

DAYS
GRAPH 4° THE EFFECT OF VARIOUS HEOIA ON

THE none or W gangamncu IN
INTERMITTENT LIGHT.

GROWTH

GOMM o

85 our» 6 - THE erreor or wmuous neon a me

80 GROWTH OF LILLIPUTIA MARGARITAGEA IN

OARKHEGG .
76

70
as
60 _ e
( queen's
55
so

45

40

HEAT
:5 ,.’I’.D.A.

.’
’1 RICE

3O HANURE

26
'20

 

IO

/——mcmno's

G 9 IO

i I

a“

 

-n—fi— 1—‘o—TT

microscopic measurements oi'spores are giv;n in 2.2913 n1 to “iii and

CRHLLS 5-5.
The size of conidia was found to vary under d' fie re;at li ht inten-

Ledia. This is shown on -_LLE XIII.
Lilliputia xergpritacea produces three types of conidia; elliptical,

ovate, and sphezical. average measurements are as follows:

elliptical 2.4-2.8 x 7.0-7.7 microns These measureaeats represent
the extreme in length and

spherical 4.5-12.7p width of each type of conidia
resulting Iron growth in

ovate 5.2-5.8 x 5.7-7.5P the media listed in T“; E XII

Elliptical canidia were most trevalent.

IV, E, 5. E'ffect of ea‘eruture and H on Growth and Re} ediction

 

To find the combined effects of varied hydrogen ion concentrations
and temperatures on growth.and reproduction of Lillipptia margaritacea,
wheat and rice agars were prepared, autoclaved, and adjum med aseptically
(15) with a Leckman pH meter to pH's 5.6, 6.0, 7.l, 7.4, and 3.0. Three
Petri plates of each medium, for each pH w re inoc dated- each with a
perithecium, and placed into incubators previously adjusted to 14, 13,
24, 26, 23, 50, 32, and.34OC. Growth measurehents were made everyday
for 10 days and fruitification results were recorded (ThBLE XIV).

Data in T.LL3 XIV w;s cor piled in the su.m*:er'of lGéB. This eXperi-
ment was duplicated in the fall of 1949, using Fries modified synthetic
agar plus .53 yeast extract in place of rice agar. as shown in THBLZS III]
and XV'the results obtained on.wheat agar in 1949 varied greatly from
those obtained on the suns arar in 19-8. This variation may have been
due to a difference in the variety of wheat used in the tvo eXperiments.

The opti.1um tamper: tures and pH's (TxbLE hfl) are based on tELe

following conditions which were considered and listed in order of their

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TAIL: XIV. Effects of -;Crogen Ion Concegtrutioi, Zea,er tare, ind
Indie on the Growth of Lilliprtiu U ri-rit'cel. (Xe sur-
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* Growth in mm. Kenn diameter of three pl;tes in euch cuse.
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importance when deterrining maese optimw:..

1) The abundance of perithecia and conidia produced.

2) The size and thickness oi‘the colony after grow h period.

5) The speed with which perithecia and conidia were produced.

4) The speed of maturation of the asci.
IV, 0. Effect of Various Factors on the Taxonomic Characters of the

Fungus

1. Nutrition, Temperature, and hydrogen Ion Concentration

Synthetic and organic media

liessurements of various fungus structures were made from cultures
grown on wheat agar and.modified Fries synthetic agar plus .55 yeast
extract. Each medium.was divided into five, 5000c. aliquots and one
aliquot adjusted to each of the following pH values: 6.0, 6.5, 7.0,
7.4, and 8.0. Approximately 15cc. of agar were poured into each of 15
Ietri dishes which were subsequently inoculated. Three dishes were
incubated at each of the following temperatures- 22, 24, 26, 23, and
30°C.

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46

'a on Tuxonomic Churecters of

Effect of_:HL_Tem9erutur 1
u nurized from Lute liesented in

Lilligutiquurreritece<
TgfiLES XVII to ELVI.

 

l

 

 

~ x ' .., ,_,_
n. i’iheqt £1ng

 

éu 0- 411 tyges of conidia become larger with an increuse in pH.

Conidiophores are largest at pH 7.0.
2400- Ketulue are longer at pH6-7.

- Conidiu absent.

0- Conidiophores are larger at pH 3.0.
5000- Conidia end peritheciu absent.

ct of Tanneruture

A
*

2 VP”
0 all

ii)

 

DH6.0- Perithecie not mature at all temperatures.
T. . l
pho.5- Asci larger at 24°C .

pH'7.0- Conidiu somewhut larger at :2 and 230C.

- Conidiophores ere somewhat smaller at 2800.

Branches somewhat longer at 2200.

pH3.0- Ho effect.
B. NOdified Fries synthetic plus .5% yeast extract seer

1. Effect 2.33.2217.
2200- No effect
24°C— Conidiophores slightly kirger at pH 5.0.
- Brunches and phiulides longer at pH 3.0.
26CC- ConidiOpnore measurements differ.
2300- Brenches, metulue and phielide measurements differ.

30°C— ConidioPhore measureuents differ.

 

l. . .
“sci could not all be measured at tne same time after me urity and there-

fore differences are expected.

2. Effect of taggereture

Conidiphores larger at 5003.
Differences in metulue end ph slides.
E0 effect.

pH 8.0- Differences in metulee measurements.

Spherical type spore not present at 26°C.

Comperison of Taxononic Chara ters on Fries and Hheet Kedium'Under

 

Different Conditions of Temgergture undng.

 

1. Effect of pH.

22°C— conidia absent in Fries.

24°C- phielides larger in wheat-pH 7.4.

2600- elliptical sgores not found in wheat- pH 6.5.
28°C— conidiOphores larger in wheet at pH 6.0.

- conidiophores lrrger in £ries at pH 7.4.
- phielides longer in Fries at pH 7.4.
no conidia end perithecis found in wheat.

2. Effect of Tenperuture

 

 

pH 6.0- branches som whet smaller in Fries.

pH 6.5- conidiophores larger at 24°C in wheat eger.

pH 7.0— conidia absent at 2“ and 24°C in fries and 2e, 23
30°C in wheat.

pH 7.4- Ketulue longer in Fries.

- branches larger in wheet at 23°C.

conidiophores larger in wheat at 24°C end 23°C then in
Fries at the sane temperature.

no spherical srores found in theet at 2800, phialides
longer in Fries at 28°C.

Ketulee and phielides lOnger in wheat at 24°C.

A?

a C 0 P1 0 7 ~ -
D. Overull Vu‘éiuti on c‘.‘ J-LiXC’CEOIAlC Cnuructers es Deteriained b3," the

 

ebo ve Exge rime nt 8

 

Conidie
elliptical 801-403 X 6. -900 p
ovoid 2.4-4.3 X 5.1-3.6 p
spherical 3.0-6.4 ’1
ConidiOphores 94.6-450 p.
Branches 2.7-6.4 X 3.6-4.3 p
I-':Ctu1.kie 2.3-5.3 X 8.6-2105 P
Phialides 1.5-3.4 X 17.6-25.8 ’1
Perithecia 400-800 p.
.--.scospores 17.2-30.1 )1
“sci 25.8-60.2 X 30-86 )1
Spines 1.2-4.9 p

49

V. D_°C-SSIUN

Then sterile whole menure is inoculated with Lilliputie merrurituces,

 

glistening, hy-line mycelium oun be seen nfter three deys growth. The
conidiel stefe is visible on the 5th dey when the myceliwn is ubundunt
but the individual strunds sepiruted. The surface of wnole menure is
covered with verious sizes of numerous, fluid, hyuline droplets just
visible to the nuked eye. These droplets are found throughout the grow-
ing area but are more numerous ut the center.

The conidioPhores are produced from a much thinner mycelium (PLATE

7
III, Figure].); they ere septete, hyeline, smooth, elmost elwuys unbrenched
end symmetricel. They average 4ll by 2O)4in size with e renge of 230-431
X 9'4'21P' The majority of conidioghorcs ere narrower ct the upper
portion then ct the bees. They give off two branches (4.3-5.5 X 25.8-54.$p)
which in turn give off 5—4 metulee (5.4-4.5 X 21.5p) which further brunch
into 2- pl: slides (2.2-3.4 X 25.8.43”) euch (PL "‘3 II, Figures 1 8c 2). --s
the culture gets older, asymmetrical brenching becomes more pronounced,
one to two brunches being pnaduced one septum below the first series of
branches.

In generel, the sterignete cut off tires types of Spores; the
majority are ellipsoidel (1.6-2.4 X 4.4-10p). Spherical (e-eP) end ovoid
forms (3.4-3.3 X 6.7-7.2g) ere also produced (PLW’E IV, Figure 1). These
spores are produced in succession and just before they ere cut off, a
fluid droplet is formed around the conidiel heed allowing the older Spores
to disperse while the younger ones lie side by side on their sides in the
droplet. r+"nis fluid droplet has tie consistency of meter until the
culture beeches Old end/or someyh t dehydruted. when the hulidity is

high, these droplets ere of vuried sizes depending on the eye of the

50

.conidiul sppurstus. Lhen.the heed becunes heavy'with spores end fluid; it
falls onto the substrate spreading the spores in the Lnnediete area.

In 7-10 days depending on the light intensity, perithecia (400-7OQp,
mostly 500flwhen mature) are produced--mostly on woody remnants in manure--
by the coiling of mycelium which produces a rcpy, waxy peritheciul well
conteining muny pupillute heirs. The Well is loose end.hurd at first
but becomes more compict and soft at maturity. The well is 6-9 cells
thick (47-75p), the brosdest cells are in the middle layer end the narrowest
on either side. Ehen first produced (6th duy) the perithecie are light
ten and sppeur pearl-like (PLJTE , Figure l), but us the sscopores mature
within the gleba (524-606)»), their color changes to light brown (st this
time the esci are fully mature end the ascospores ere hyeline and immature).
The mature perithecie finally appear rust colored due to the mature rust
colored escospores (19.5-21.5p) (FLATE V, Figure 2) showing through the
still light brown peritheciel well. In about 15 deys the esci appear.

They are pyriform and loosely packed, but just before disintegration
(17-19th dey) of the escul well the aculeute, hyuline sscospores are
tightly peeked (PLJZE V, Figure 3) within the escus. The mnture asci are
difficult to see because of the thinnes of their Wells. They ere bulg-
ing (24.4-30.1 by 36.5-55.9»), end heve reuched their maximum elusticity.
Upon disintegration of the usci, the sscospores are still hyuline, and
beer aculeste spines shout é the diumeter of the spore in length (PLgTE
3, Figure 2). The ascospores ct maturity becoue verrucose to echinulute
(1.9-2.2») PLJTE 5, Figure 2.

Upon comparing the cheracteristics of this fungus with the descriptions
of those listed in the appendix, it was found that our-fungus resembles

three known species. The similarities and differences between Lillieutie

51

geillhrdii, Gliocl:-..dium prolificuml, Lilliput in insignis end our fungus ere

 

 

 

discussed below.
The mature esci of Lilliputie g-gilerii are described (5) es oblong.
The escospores are light brown at maturity, but eppeer white when seen

through the per ithecium. ‘i'he nature escospores of Lilliputie HIJIQ‘LAI‘l'tLiCGLL

 

are el':-.eys rust colored at rmturity independent of the type of media used

H}

for growth. *he mature asci ere subglobose. The esci of Lilliput is

 

Eilllbl‘dii as shown by Boudier end Petouillerd (5) are loosely packed

 

comgnred to those of Lilliputie ::i~;r§:.iriticea (P1423 5, figure 3). The

 

esci of Lilliputie guillerdii are SIiLll compared to those of our fungus;

 

ascospores of both species have a "warty eppeerence".
In the description of Glioclediwn prolificum, the conidiophores
are described as being brenciied at every septum. This cinrecteristic

is not ordinerily exhibited by our fungus. The conidia of Lilliputiefi

murgsritucee are mastly elliptical. Those of Gliocladium prolificum

 

 

are mostly oval. Perithecie essoci-eted with _C_}__._prolificum were thought

 

to belong to the Perisporieceee, however from the description of both

stag-es (2) that species appears to be congeneric with If. :rn-rgeritecee.

 

The color of the polyhedral cells in the peritheciel Well, the size
and color of the sscospores and the size of the echinuletions =ire like
those characteristics of our fungus.

Dennis and :iekefield (9) (bel ieve thht _L_,_ insignis, 33:. insigne, _I_,_:_

geill-erdii, endgl penicillioides are probebly the some species. The

 

fungus L. insi ,crnis as found in march 1933 at Uxbridge, iniddliesex, is
described es foll we; fruitifice ;io:1 glebrous, sp;_;ericel, white to

ere-em colored, approxigzetely 500).). in diameter, and huving e stout Well.

 

. C -
These c-11recteristics comeide Wl‘th those of L. mergerituceu, but the

(51
m

. I
escospores ere s ell (ll-l9) Congsred to those of our fungus (19.b-;1.§P).
he m-asurerevts of L. insifnis were nude of yellow escosgores within asci
end these spores are studded with short blunt hurts. The sscespores of

-

L. m rwuritscee_w'thin esci ere colorless end have spines 3 the diameter

 

Q.

of the spore in length. ri'he Spores ere durk brown end hive short blunt
Werts only when muture end the asci disintegrated.
; culture of'the type Species Glioclediu: penicillioides the obtained

a C
from th "Centrul bureau voor Shimmelcultures" on cherry user for comperison V”
,

 

with L. mirrurituceg end no fruiting Wes found. vhrious media here pre-

 

pired end inoculeted but nothing more then sterile mycelium hed been
formed.

A vuriety of synthetic und orgunic media were prepured end inoculeted

 

with L;_mergprituceu in en attempt to produce both stuges of the fungus.
0f the synthetic media, Fries modified synthetic (plus 5Q yeast extract)
produced both stages of he fungus, and peritheciu netured within ll duys.
Further experinentetion is necess:ry here to produce a true synthetic
ever. an epproximete.unelysis of-Decto Yeast Extruct Wes obtained end
‘howed 29 known compounds contuined in yesst extract which included Vitu-
mins, unino acids end inorganic nitrosenous coupounds. These compounds
should be broken down into snuller groups and then by s process of
eliminstion, the compound or compounds which end to the growth and re-
production of the fungus, would be known.

Enngi are clessified into four rroups on the besis of“the form of
nitrogen which they ere eble to utilize (39).

l. Utilization of orrsnic nitr03en

2. Utilization of organic und emmonie nitrogen
5. Utilizution of orgenic, unmonie end nitrate nitrogen

4. Copuble of fixing elementul nitrogen und utilizing ell other
forms.

The form of nitroren necessury for ussimilution therefore, depends
on the netutolic potentiulities of the ergonism. nn attempt to produce
a synthetic medium, using Coons egur es o buse und substituting vurying
molsrities of supprs in combinution with two nitroEen sources (hsving tie
sums equivulent of nitrogen), failed to produce both stuges of the fungus.
Conidiu were produced only in the nnltose-u-sspurugine conbixutions nnd
no perithecis were forned. Led more time been nvuiluble, here too,
nutrients such es vitumins and emino acids could huve been added. gorever,
fries modified synthetic (plus .Sfi yeast extract) produced both s+uyes of
the fu ngus .

Eecuuse fries modified synthetic (plus .55 yeust extrsct) produced
better growth and reproduction thun any other synthetic medium prepured,
it nus used in e study to show the effects of environmentul cnnnges.

wheut mes also chosen to represent on organic medium. Fries modified

()1

synthetic (plus . fl yeast extract) on the whole produced more fruitificution
and better growth than did Wheat.
Ieusurements were nsde o; verious fungus structures from cultures

grown on wheet and Fries modified syn hetic (plus .53 yeust extract) which
were exposed to Vurious temperutures and pi's. Results showed that of the
structures meesured, the mature ascospores end spines were the only
structures which nge consistent measurements independent oi‘the type of
medium used, p3, or temperature. One would deduce then thut in this group

of fungi (Lilliputiu), measurements of ascospores and spines, tould be

 

most inxnxtunt in classifiCution and identification.

RA

PLACE I

~I5‘—.—‘

 

 

 

 

~~~' ,.- v ‘u n ‘wv n-at xioo
Fit—"111‘s 1.,L! .~..: i‘LiJ.IC‘i1 0-- I1 LIL l.I_-..[ l

 

 

 

‘ ‘ " "s‘ =0 r~* 1“ iLSQENDIH
Figure 2. Gthlltnglou on An u-oo- one (no,
’ ‘ X100

55

 

 

 

 

 

l o CUi: IT’IAL :Ixi’} 9-}-..5'1 le ": X150

 

 

ricure

20 :Olzi'JI-L]J til-“\xiggllflkjif 1:15»)

56

 

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58

 

 

 

 

 

 

 

 

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59

O
r ((-mw‘ gar “‘1' fi“"./TT“"_“T"""'
‘«I. an--. i .L ‘. J 'VK.‘ Jp'4._).‘..v..o

1. This fungus Wes fcund in Lust Lansing, Lichigun by Dr. Ernest
Bessey, in the Eull of 1944 in some stuble nunure.
3. Single cell isolutions of condidu end oscospores were mine

end gerninutions of both types of spores produced an asexuul stuge belong-

ing to the genus Gliocledium and e sexuul stuge of the genus Lilliputiu.

 

3. A culture, resulting from u single ascospore germinution, nus
grown 32_Vitro und used to determine a) optimum tenpereture, b) suituble
eurbon and nitr03en sources, c) optimum hydrogen ion concentrution, d)
possible synthetic mediu which could be used to produce both stuges of
the fungus, e) growth.on v rious organic media, f) the effect of light,
intermfttent light, and derkness on growth, conidiul meusurenents, end
fruitificetion of the fungus and, g) the combined eff ct of veried
tenperetures, pH's and media on growth, fruitificstion and morphology.

4. In generul, orgunic nediu were quite suituble for growth und
reproduction of ;=_nnrggrituceu in tnet both stuges were produced in
abundunce in a short time (10 days).

5. In an attempt to produce a synthetic nedinm.on thich the fungus
produces both stages, vuried molsrities of sugurs were used in combination
with two nitrogen sources (K303 and e-espursgine). The conbinetion of
.03 molur multose end e-ueperogine resulted in conidiul formution. Io
fruiting was found in other coibinutions.

6. Fries and wheat ugurs were used in an experiment showing the
effects of temperuture, media and pH on the growth and meusurements of

the fungus. The following conclusions were druun from the results

_obtein863

u) Veristion in pH at s constant tenperuture cnuses upgreciuble
differences in messurements of conidiophores, ccnidie, end metulde.
Other structures are fairly constant.

Fries modified synthetic (plus .5; yeust extruct);

a) 'Vuriution in pH at a constant tang ruture causes upgrecieble
differences in meesurenents of conidioghores, bruncnes, phislides end
netulee.

7. Fungus growth occurred from 14-5000, the optimum temgersture
being 2400. The ph optimum differs with media used.

3. The effect of ligh on fruiting Wes influenced by the media
employed. On most mediu, however, perithecis and conidia were produced
1“ greuter abundance in the ubserce of light.

9. 30 effect of light upon the production of the three types of
conidia found could be sscertuined.

10. Optimum pH and temperuture for sporulstion were determined
to be as follows;

On whect dyer,

1. For conidisl production- pH 3.0 at 27°C.
2. For gerithecisl production— pH 7.4 ut 24°C.
3. For both conidiul and peritheciul groduction- pH 7.4 st

0
C.

24
On Fries user
1. For conidiul production- pH 7.4 st 5000.
2. For peritheciul production- pH 6.5 st 3400.
b. for both conidiel end peritheeisl productios-ph.6.5 st

2403.

ll. Size of conidiophores, spores find metulce Wes sporeciubly

. V" ) ~ ‘ , o - .
altered by u chines in ph oi sheet spur. ot;er cuurccteristics were not

1:. bias of conidiophores, conidiOQhore bruncges, phislides, end
metulse Wus effected by u change in pi of fries synthetic (plus .53 yeust
extract) medium. Other charecteristics were not effected.

15. Size of conidiOphores, breaches, getulue, phiulides, and
perithecie Wes found to be effected by the media used.

14. The Variubility of the tsxonomic cauructers of tge fungus
under considerutfon, under different environmentul fictors Leving been
determined, a comgsrison of this fungus with the descriptions of

Glioclidium_prolificun, Lilliputis feillurdii, Lilliputi: insignis,---

 

three known species which beer close resenblence to our fun:us-—-leud
to the conclusion thst our fungus is a hitherto undescribed species for

which the nume.Lill'putiu nurseriteces is proposed. n forhul description

 

is appended.

62

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Glioclsdi1m_p9nicillioides Cords. lBéC. Icones Fun oruu 1.. 5C. fl.

 

’7’ i’il’ . 9:2“.

- » n ,-. A. r" x / '5 1"
notrucnot. ILvU. hevue G5”of l he sotinioue
Penicillus clusters very snull, punctiiorn, white stipe erect,
flfilkuois, wary, thickened above, septate, powdery white, brunches '/
opposite, brnrchlets verticillste und in fours, crowded. Eecd of

-"lutinute, oblong, bound by c

O
"J

spores globose, white, spores c
thick gelutinous lays . Eech spore hes u thick geletinous luyer.

Length of spore 5‘p.

71

w

helephoru hirsutu-und

(L
(D
Q

d»:
P
C3

C Q
I

Grows on the hymenium of
o /
Th. 5 nfuinjlentu.
The whole plunt ranches l/lO-l/d of u line (.1 inch). Clusters
ere visible with only c sharp eye us tiey ore smell white points.
They rurely reuch the disaster of l/Smu.- every cluster consists of

.L

15 or 20 conidioohores thich grow upWerd end twine. They grow up
winding together und liter upright. Under low msgnificution, stalks
uppeur like thresds, club shsged ut the top which beur spore heads.
Under the high power, consists of branches end brunchlets tiich heve
the s;ore heed on top und the end brunches producing the slores. The
Spores are very smell, oblong, end somewhet unegusl und obtuse. They
ere white and surrounded by u clesr'xhite thick slime lsyer. The
stipe end its breaches ire divided und trunspurent-nhite, hollow with

rou hnesses externully. Jrunches ;*d brurchlets ure uttsched just

63

J
yosite - the thin branches

below the septum. The thick branches are 0p

and lust brunches ere thin, whorled end in fours.

Glioclsdium lignicolum Grove. 1336. Kew or noteworthy fungi. Jour. Bot.

 

A Gliocludium with gregerious or fdéfiCUlute hyphse, erect,
egunl thickness, 3-4 septnte, hynline, at apex penicilletely di-
trichotomous; heed of conidiophores globose or obov;te, white;
conidia stuck together in mucous, hyeline, oblong-ovoid 21-23~ by 135).}.
fies found on the surface of wood lug. to Sept. Does not differ

n

from G. benicillioides except by the conidia being-g as long, and

 

its habitat. Perhaps it would be better to consider it a Variety.
So etimes the hyphue in the lower dzy'ynrt oi‘the wood are 5 branched

and colored brown.

Eurotium insigne hinter. 1337. Rubenhorst. Kryptossmenfloru von

&

 

Deutschlsnd, Uesterreich und der Lchneiz. steite “ufluge 1 (2):
61.
Kycelium spurse. Peritheciu superficiel, round, wnxy, yellow,
soft, m.mbrenous, smooth - .35 -.44m. “sci ure irregular, oblong,
\/

contuining S spores. “sci disintegrated quickly and are 55-4dp

long by ES-EOp wide. Spores are dressed together, roundish, almost

. . i . . . A r 3
colorless and heuv1ly covered Wltn snort spines le-lop in length. x
This culture Wes collected from goose manure from holle. V

('1

lhe author nentions conidia present but no churucteristics ere

given. ”one possible relction to 5. genicillioides is indiCnted.

 

Glioclsdium co:;;ctum Cooke et Hussee. 1337. Sens qutiC Eun;i.

 

F.

urevilleu. 15: lb.

 

Clusters minute, punctiform, rust color. hyphce erect, crowded,
fonzing compcct clisters, septcte, mostly simple; heed of conidia
cuneute, pcle smoky, long, covered with mucous; conidie stuck together,

oblong and in chains 5 by hp, an ny-hysline.

Found on paper from India.

Letruchot. 1395. Rev. Gen. not. 7: 431.

In chrrot, color Veried fron unite to orange. Jertile brunches
short, slender, end apprdxiietely ZOOF high, hp in dismeter, hardly
larger than the creeping mycelium. in the loner gurt of the culture
tube, spore heads ere so abundant thut together they give the uppeur-
unce of s unifonn mucilsginous muss covering the substrutum. In
old cultures the orsnge color renuins indefinitely.

Spores ere 4-5 by 2-3r.

Gliocludium ecuricinum Cooke et Xcssee. 1339. New British fungi.

 

Grevillea 17: 30. I

Causing the pileus of mushrooms to crack into large frustuler
scales.

Tufts hemisphericul, soretines confluen , pullid, gr wing white
at first gelatinous. hyphue creeping, branched, fertile, erect
brandzes producing ultihcte verticillnte quaternute, cepitulum of
conidia-subglobose, white, hysline, 5-dP in disaster.

Found on cultiVuted mushrooms, Leicester.

Glioclcdium viride fistruchot. 1:93. bull. Sec. lye. Erirce 9: 243-352.

 

This Species was found on Clitocybe; collected in s woods in

65

France in April. Appeared in the fonn of smell green droplets of
muciluginous substance supported on short conidiophores.

Once collected, §:_viride was gromn on carrots, turnips, gelatin,
etc. i; Germinuted spore gives rether Vuriuble mycelium from 3-qh;
septete, branched,'nith snellings here end.there.

The diameter of the conidiophore enlirges sl wly up to 12? end
the end becomes spore beuring. branches of several sizes S-qh to
B-SP. Mt the end of euch'brush, there are first distributed ovul
spores arranged in chains and for so shine head like. These spores V
secrete sticky meterisl that envelope the spores into a speriCel
green mess.

v~

ranching of the conidiophore occurs producing new brunches

U

directed downturd and completely giving a distinct root like or

anchoring effect.

T‘f‘

Gliocludium necropodinum E. Ehrchcl. es . Chumpignons Coprophiles de

 

Belgique. Bulletin de 3 sec. Roynle de notunicue de Belqioue.

 

 

4;: 135.

0'1

Clusters white, delicate, sterile hyphee slender, flexuous,
creeping, fertile ones erect, strong, 3-4 septute, ESQ-ech by 14-
ldP bearing u muciluqinouS'nhite little heed, the besidiu several,
conidia cenglutinete, hynline, 9-11P by B-ELP, oblong, continuous
with epispore smooth and guttulute.

On dung of 'unguroo in Belgian.

Lilliputig guillurdii Boud. et Put. lQOO. Bull. 30c. Lye. Er nce 1&5-146.

 

 

This smell species is herdly visible to the neked eye. It is

seen as small whitish grains which have not been seen with mycelium,

end ure disgersed in the humus of used burk us those of Coccobotrys
end Xylopilus were found. The author nentions its resemblunce to
Terfeziu but is well distinguished not only becuuse of e difference
in width but ulso bec use of the composition of the peridium. The
corticul layer reuches about the middle of the central portion an
is 35-90p thick. The corticel luyer is nude up of polygonul cells
which are ruther lurge in the centrnl rerion, end sulller end more
granulur et the surfuce end n ur the glebu. It is e little more
colored then the glebu which is entirely white und seems to be
forged of jelly-like mutcriul in which the esci ere oblong without
the uygeurunce of busilur enlsrgnent which hey be there end ere
LE-Sep.in diumeter which contuing 3 spores, seldom less, und which
reseuble Terfziu in size end werty upgeurence. These uscospores seem
to dive oily drops and st :hturity are light colored but visible.

The retire SIOTGS ere light brown end look white when seen
through the perithecium. “5 shown in plate V, esci which enclose
mntu escospores are loose.

The perithecie are not over-g mm.

Gliocludium.hgponzces Succ. 1907. Srll. 4: 30.
Sums us Eenicillium hygonyces.
Scattered white, hyphue; erect septete, hyeline, with erect
apex, cupitute. Di-tdtri-chotomous, Conidiu cutenulute, ellipsoidal, V

hyuline 5-4 by 2?” 0n Stereum hirsutum in England. Close to P.

 

eundidum from which it seems to differ by its ellipsoidul conidia

end its more regular branches.

RV

Gliocludium roseum Beinier. 1907. Eul. Soc. Lye. Frunce 23: ill-112

 

Flute 15. fig. 1-6.

Conidie: two conidiel appearances-colorless to pink, in
nnss, 5’7P by 3-5u, slightly epiculete. On potuto dextrose agar,
colony is white to pink or selnnn in fruiting areas which ere pro-
duced at first, the hyphse being in repes. Dense pinkish masses are
produced.

Kycelium: branched and septete.

ConidiOphores: iconsist of more or less elongated support aris-
ing from a single ternunul verticil of 3-6 long sterigmetu. Eech
sterigme produces a series of conidiu cOuted with u gelatinous sub-
stunce pnaducing one spore after unot er. These conidia do not
remuin together on c sins. Right after forming they slip successively
on conidiu underneuth to which they stick forming e large spherical
muss. Iormul conidiOphores are pnaduced much lster end soon cover
the total surfuce of the myceliul muss. These conidia give a rose
coloration. Conidiophes send out one or two lsterel brenches which
in turn form 1 or three snull branches which brunch out eguin or
terminate in 3 or 6 short sterigmutu.

This fungus hes the property of producing two fruiting systems.
The first has already been discussed, end the second; long cylindrical
branches of conidia which rennin united, rd lie side by side and

pL'ILllel o

Gliocludiun pulchellum.Penzig et Succurdo. 1909. Syll. 13: 52l.

 

Loosely gregarious, hyphee fertile erect, simple, continuous
(not septute) white, 1 mm., tall, apex not infleted, base 4O‘SQP

- thick, Reeds globose, hemispheriCel, dirty rose colored. Besidie

very dense, pencillately radiating, forked or simple, filiform,
SO-9Qp by 1.7-2p, conglutinated by mucous; conidia minute, ovate-
ellipsoidal, in chains, 2.3-3 by 1.7—;p, pale.

Habitat; on thalli of lichens, on living shoots and leaves of
0 V’
mosses and probably parasitic. Tjib‘das, Java- very distinct on

account of non septate hyphae. Probably type of a new genus to be

called Cladoglium.

Gliocladium luteolum Von Prof. Dr. Erenz V. hohnel. 1905. Lycoloeische

 

Fragments. hnn. $19010: 9' 523'

..J

 

Sterile hyphae obsolete-fertile soattered, smooth, dilutely
ochraceous, hyaline above and thinly tuniCate about du thick; below
more thickly Walled about 10p thick-about 5 septate-SdOP tall, peni-
cillately'brwmched above. The branches verticithe and these again
branched. Crowded parallely spores on apiCal branches, conglutinate
into a globose yellowish head SO-lOOp thick-very'numerous spores,
hyaline oblong continuous hyphae 5-7 by'SP. Collected scattered on
I“: c

dottdn wood in forest at tassergespringe, hienerwald, lower nustriu

in Oct. 1903.

Gliocladium.nicoti3nae Oudemans. 1909. Saccardo 18: 521.

 

Sessile creeping hyphae, a little thickened at the base hyaline-
fertile hyphae, erect septate, hyaline, branched. Primary branches
quaternate-verticillately, crowded, cylindrical, 32-43P long; secondary
branchlets turnate, basidia arising 2 from each secondary branch,
cylindrical, hyaline ldp long. Conidia oblong, hyaline, continuous,
S-lOP by 5-4p and conglutinste into a head fonning a gelatinous drop-

let 11mm. in diameter.

6.

(4.x . \/

0n decaying leaves of’ficotine tebnci, nmerongen, iolland

 

(collected by C. J. Honing).

Gliocludium elatum_Succ. 1909. Ann. Eycol. 7: 434.

 

 

Hycelium white, cottony, loose, shortly sprend out. Creeping
sterile hyphae few-conidiOphores all erect, long, 1500 by é-qp fili-
form, septate, capitate at apex, heads globose OVate, interior mucous
for a long while firndy'conglutinate and under lens opaque, but
soluble in glacial acetic acid. rseudo-basidia nice verticillate,
primary and secondary fasciculately turnate or quaternate. Conidia
catenulate, Shall, subglobose, smooth 5.5-3 by 2.5:‘hyaline.

In old wet partially deCayed pilei of Schizophyllum commun

(Al: F " I t, -
in green houses of the Eotaniét garden at rashes, “pril 1909.

Close to Glio. hypomyces but differs by its conidiOphores 4-5 times

 

longer although na rower, by its spheriCal conidia and by growing

on SchiZOphyllum instead of Stereum.

Gliocladium prolificum Bainier. 1910. Bull. de Soc. 23001. 26: 335-339.

 

 

Conidiopiores; branching at practicnlly every sep um. Conidial
apparatus consists of two layers of 3-6 verticillate branches which
in turn consist of one or txo cells. each upper branch has 5-6
sterigmata-dilated at the base and becoming smaller at the tip.
Length 6-9 times the lurg st diaaeter of‘the base.

Conidia; irregular and unequal in size, producing he below the
b/

. . v , . . .v’
other, sons round but most of t18£ oval; 4.h by b.1p. a sown conidia

produces a conidiophore apparatus, horizontal hyphae, conidiOphores

70

with a dianeter more than 9.61p.and produces a Quantity of perithecia.

Conidia for ed, gelatify their most external menbrane and slide
one over the other to fern a regulnr spheriCal uses of viriable
diuheters. pfter a while depending on temperature (15 days to three

:1. ./
weeks) a net nork is formed from which is derived perithecia in a
spherical mass of vgriable dimensions which can attain one mm. or
Amore at maturity. nsci appear later containing 8 spores.

Eerithecia; composed of large polyhedral cells of coffee with
milk color. In the internal ca' vity, asci are borne on short filaments,
in the fonn of an irregular, oval see which disintegrates releasing
spherical spores 25‘2%P covered with a membrane ornamented with small
protruberances which are isolated but many times united. The height
of these motruberar ces mea su es %P° Spore color is yellow-brown.
Upon germination mycelium is produced resenbling that of conidia
but does not gernnnate as fast. This Was thought to belong to

the Ferisporiaceae.

Gliocladium piliforte (Pers) Boud. 1905—1910. Icones fiycolosicae ou

 

 

&
ICOEOLFV““18,A~S Ch-:pi nons Us irance. E. Boudier. Tone IV Planche

537. 546-547.

This small species is composed of nunerous filaments united in
colonies, erect and ending at the peak by a small rounded white head
formed by the united spores on he top of a small cupitulum. These
filaments are rigid but nevertheless are a little flexible, simple
in all their length e: ccept -t the extreme peak there they a: e divided

e“a 11 penicillifonn branches; they are septate, blackish when seen

thru the magnifying glass but are rust under the microscope, divided

71

at the peak in 2 or 4 very short branches, evenly colored which
support other uncolored (hyaline) cylindriCul divisions; these are
then divided at their p uk in 4 others, these supporting elongated
sterigmata which pnaduce the conidiospores. The whole is held
together by a Water soluble mucilage in a globose head which seems
supported at the base by the 2-4 snail colored branches. The spores
are hyaline, elliptical nithout granulations and are 3-%p long by
3;-5p wide.

This snall species is.found on rotten wood, where one does not
see it except by the aid of u magnifying glass as smell blhck bristles
surmounted by s smell white head. Exnmples shown hnve been recovered

in February on the wood of'a deed poplar.

Gliocladiwn deliguesceng Pinkerton. 1936. nnn Le. Bo . Garden 23: 47.

 

 

Colony; at first colorless, of meinly submerged hyphae quickly
becoming thin and white-floccose over the surface then meadow-green.
Slimy on carrot plug and blackish green on P.D.n.

Conidiophores; two types— penicillate and simple cephalosporial
becoming green and aggregate in e heavy slime, Iriseriate and each
series trichotomous; brush 45 by 4QF; first series 17-24 by in;

second 9—11 by 4p; phialides 6.9 by 2—3.5p, spores 3.5 by 2.5p.

Sopp, Olav Johan-Olsen. 1912. honographie der

 

 

.v;
pilze-gruppe Penicihium, Videnskupselskepet Skrifter I. Lethemstisk-

natur-videnskabelig klasse 1912: 89-93. V/{

dhen atmosphere is dnnp and the fungus well nourished, conidia

1 . 1 ~ . " ‘—' ' v \ q
flow together in‘g Oily, a syrupy, diiiicultly floting liquid which

‘1

finally sinks whole fungus.

V

unile attached to sterigmnta the conidia are pointed at both
ends but later are sonnwhat rounded.

Hub. Found growing on the upper side of Daedalea unicolor.

 

Thom.C. &,K. Roper. 1949. Kennel g: the
Penicillin. fiilliams a "ilkins Co. pg. 630-631.

Penicilliun-like conidiophore which luter becomes enveloped in
slime nesses us the conidiul chuins dissolve and run torether;
reverse gray at first, later dark green, almost black, odor charac-
teristic; gelatin linuified; conidiOphores up to 1 mm. long, erect,
course, septste, 1-5 times penicillute branching. Conidia about
le.5-2.Qu or somewhat larger When ripe, at first fusiform, later
becoming more rounded at the ends, produced in chains which break
up us the conidia become enveloped in musses of slime; perithecia

and sclerotia not found.

Gliocladium africanum Eichelb. 1913. SuCCurQQ 22: 1279.

 

Solitary, scattered, with sterile hyphae penetrating the sub-
stratum. Conidiophores erect 170-870-600 by 3-9p4 The apex not
thickened, faintly yellowish, remotely septute, paler above,
penicillutely branched.with loser whorls in 5's. branches closely
crosded, conidia acrogenous, solitary together with brunches and
branchlets surrounded with mucous, hyuline, smooth, ovoid 4 by 2p.

Hub. On deCaying wood, oblong with Tilmudoche nutens, nmani,

 

East nfrica.

I/

. -. it , a . . x . - » . .
GllOClLfllum cinereumlnurcnul. 1941. bontributions u 1'etune nee chumpignons

 

I/

,0.

f
fructicoles de belgique. hul. Soc. oy. Lot. nelg. 54: 150.

 

 

Clusters dense snort, ushy with sterile hyohue creeping and t

-.l)1

copiously septete, about 7? thick. conidioyhores, 5-4 segtete, erect
or ascending, simple, 5-4 tines verticilletely branched. Iroducing
d ty;e of brush O-60p by 5i-5.Zp. Conidis globose, somewhut ovoid,
one drop of oil in eedh conidium, 3-0‘5-fifl in diuneter surrourded by
thick mucous.

Heb. Cn peer fruit (Gembluux)

Observ;tion- By its smeller sgeres and by its color, it is
cleurly distinct from other sgecies. ’

V’s-“W /

This species Wes found on peer of the Zeurref It entirely

covered the mumx fied fruit with its ssh colored guuze. Col n

  
 

 

 

 

chlamydospores l5 by 15.5p, terminnl 5 by 11p; skinetes 15p in ;}//
dimneter; arthrospores 10.5 by 5p; oidiu 5 by 5.5p, rure.
\‘_’flfl;ihb. Peresitic or semi-puresitic on certuin species of greed.
Gliocludium finbriutum Gillmhn and nblott. Cot. lszd-July 1327. .Lgii
State 031 e;§_Journul._§_5cierce l (5): 302, 504.
On Czupek's ugur; colony is pure white at first w‘th zones of
durk green fruiting dress upgeuring at the center of the colony.
Conidiophores arise from seriul hyphse, smooth end up to Sou long.
Reeds ere enveloped in round bells of slime in mhich chuins are not
D-

distinguishable. fruitificetion is in tho stsges, breaches end hetulue.
Conidiu mny be born directly on phiulides which arise directly from

the conidiOphore. In most heads branches erise sons disturce below

the mein heed. Phiulides lO—ZQu long. Conidiu ellipticul or elongate,
ovoid, smooth, pale green, 6.5 to 9.5p by 3.5 to in. "From soil: U. S:

Ions, Louisiana."

74

r/

(D.

 

 

 

 

G1iocl:§1:n dtrrn ”illuun end “bbott. Oct. lQSO-July 1927. Ion; Stut
Collere Journul f Solence. l (5): 503, 505.
Colonies on Czepekis u er. brown-green, smell, slotly snreud-

v"
in;, lurgely submerged, eerisl mycelium olivsceuous, scunty, eeriel
erowth consisting mostly of conidiophores; colonies moist with sline

which envelopes the heuds. On been ugur considernble mycelium is
bonidiophores urise es oliVuceous, thick welled, smooth,
c r”
truitifiCetions ere in three stuges,
V

produced.
und septite, 75-500 by 5-eu.
Erenches 3.5-9.5 by 5-5.5h; metulue
Conidiu OVul

lides 7.5-10 by 1.5-2.5p.

soznetis;es in t°.~'-zo or fbur.
3p; p113.

oblong, 7.5-9.5 by

to ovoid, 2.5-4p by 2-3.5p.

U. S. Louisisns.

IOWu
—

From soil.
Oct. lOEo-July 19;7.

 

I'ILL b o
Glioclsdium cutenule+ i Gillmun.end nbbott.
Stute Colleye Journil of Science. 1 (5): 502, 505. Fig. 57.
; pure white becoming olive green to bright green
...L

 

Concentric zones sepureted by sterile
llowish. “eriel mycelium

‘73

“V

cm Czepel-z' s usur
in center as fruiting ureus develop- fruiting dress were cortined to

the center of the colony.
Volor reverses from colorless to

myceliun.
abundent, simple or in ropes from.nhich ccnidiophores erise.
ConidiOphores once or twice branched, course, rough, pitted,

50-125u long.
C,
'ruitificetion in 5 stages; elenents of fruitificutions pitted
Eriniry brunches 15-20 by 5.5-eh, metulue 7-9 by 15-25P,

.r
.4

"7 S-épo

or rough.
phielides lO-ZCF.
Conidiu ellipticul, smooth, pnle green 4-7.5 b,

Isoluted from soil.

Glioclrdium microsporium Fetch, T.- 1935-27. ldditions to Ceylon fungi.

 

Ceylon Journul f Science. 10: 157.

 

 

ihite, forming sub-cylindric or syre ding fuscioles, up to
.dmm. hi:h, .lmm. din eter ut the buse; conidiophores brenched neur
the base, simple above, éu diuneter below, slightly uttenueted up-
Wsrds, 5n dimneter ebove, hyeline, septute, struight, sogewhit rigid,
closely en‘ minutely verrucose, beur'ng at the apex a few, usuelly
4, cylindriCsl proyhielides, curved below, 5h long, 5p in diameter,
each beuring two to four elongated flush shuped phiulides, a-ldp
long, 1.5p below, the prophiulides end phiulides forming u lunceolute
heed, 15-24 by 3-1Cu; conidia edhering in a globose mess up to 50p
in diumeter; conidia hyeline, oblong-ovul, 1.5-5 by .75-1p or sub-

globose, 1p in diameter.

/"W .
On Folystictus flebelli£formis. V/
us

 

Gliocledimn fLivum'Vun Beymu Thee Kingma. 1933-50. Ueber zwei von

v n, /

Hepeu-Rinde isolierten Pilze eus Qumitru,'&ittei1ung eus dem

 

A ./
"Centruel dureeu voor Schihmelcultures". Verhundel+ingen der Kon-

 

‘

inklijke hkudemie Van te.)etenschupuen, “meterdln, (Tweede Sectie). EU.

in.— A;

 

Clusters en melt—beer ugur dsrk yellOV-woolly proding in zones

and ejreuding rupidly— underside orunge yellow. hyphue-sterile-
creeping, fertile rising- fertile 2-5 mm. high- hgnline septute-
4p brOed. Conidiophores not ubundunt, smooth, septute, irregular
going off right and left from.mycelium. SC-QQF long, 5-4p breed-
1-5 brunches- every brunch at tip with 5-6 sterignutu. The latter
\are elonnuted, bottle shsped, lé-ldp long by 1-.7p broed- nurrow

v

to‘imrds upper end. i.-.uny times the sterigzlutu ere surrounded by slime

76

end form in his cuss spheres of lO-l5p.inhdisneter out of which

touching with gloss ncedle, greet misses of spores spregd out.

Conidid beun shuped to ellipsoid, snwoth, hyuline, 4.5-9.7p long

(mostly 4.7-6.7p.meen of 100 spores 6.02p) und 2.5-4.7Fbroud.

Clostly 2.7-5.7u meun oi'lco spores 5.85p). The spores germinste

eusily in filter in the process of swelling to double their size.
Pure cult ur e 5:

On rice- after 7 duys becomes sulfur yellow but st upper
end of tube little white mycelium.

On huulin egur- after 14 days, thin, slightly sooly skin,
greenish shite.

On Out melt end Corn meul- ufter 14 duys- ulmoet nothing
grown but on surfece, numerous Snell drOps of tuter- on corn meul
a few small orunge yellow colonies.

403 SeCCurOSe- ufter 7 days, thin skin. The inoculuted
pluces ere firsnulur,'whitish yellow end utter 14 days orange-yellow.

Currot- after 7 days strongly redieting mycelium, after
14 days becoxes yellow but currot not entirely covered.

Potsto- in 14 days, white wooly mycelium, but pieces not
entirely covered.

Found on burk of Keven (Indie Rubber tree) Sumutru.

Glioclgdium nigro—virescens Vun Begun Thoe Kingne. 1351. Ueber ein

q,
‘4

 

neues Gliocludium. Litteilungen udS den "Ce truel Jureuu voor

 

 

Schimmelcultures". III. Verhundelingen der Koninklijke “kudenie
Vun Tetenschuppen ts “gsterdun. Theede Sectie. Leel £9: *0. 2:

 

30—52, fig. 1.

fiycelium on beer-wort ugur in u :etri dish not vigorous, sale;

'7'?

What crisp woolly. Lostly consisting of bundles of hyphue st first
greyish White tien lirht green shout 547 in Klincksieck and Vulette
Code of Colors: growing in severul zones which after severul weeks
become derker by conidisl fonistion (542-575. K a V). nt a greuter
distance from the point of inoculation, the mycelium is altuys more
spurse but on the contrary the messes of conidia ere more numerous

so thut the outer zones consist only of’conidiophores. The murginsl
zone is forxed of myceliuu lying flut on the egsr, grey or somewhat

-‘~

reddish (Ben), he underside oiPthe petri dish is yellow (3334-211-

\—

06). To typical oébr. Under the microscope, mycelium,shows numerous

Q.

oil roplets. ConidiOphores branched or unbrunched, smooth, 40-100p
long by S—Qu, greenish with numerous oil droplets. ‘Ietulne 15-25 by
2.7g.

Thielides- flush shaped often with long neck, 20—55 by 5.Zu.

Conidiu— oblong or ellipsoidul, often somethut pointed at one
end. boxetines been shuped or more rounded. Smooth, greenish,
singly, in mass: black-green with l or 2 indistinctly visible durker
spots in interior. united in greut nnsses mostly in slime heads
15-70p in diameter.

In pure culture; after 7 duys at room temperature

Beer Khlt ugur; little loose woolly white mvcelium over a

greyish bleck:esur surface due to conidiul formation. 350-

875. Reverse yellow 211.

Currot- no eeriul mycelium. Conidiophore heed covered with
conidia for most pert spursely, but in lurger numbers on the sides.
Burk green.

Potato- overgrown with vigorous develOped but little aeriel

mycelium. Violet above (5051) then follows a zone with muny conidia

375 while loser dean fungus grows leutnery.

Hub. In g rden soil Buurn, fiollund.

Glioclidium Vermoeseni (Biourre). Pinkerton. 1956. nnr.1s ££_ he

 

ngSOdTl Botinic;1 Gurden. 33: 47-43.

T»!

 

L e
Colony: pule vgnucious, gro tn repid procucing much guttution

tutor.

ConidiOphores: mostly benicillute mcesuring 4gp from the first
brunch, asymmetric, polyseriute with the phielides trichotomous end
meosuring up to 10 by 1.5-5.5p, spheriCul s_ores s.sp; cephelolides
40p und more, cephalosporie aO-abp, ceghnlospores ellipsoid 5-6.6
by fip; interCLlery chlumydospores 13 by 15.5p, teIminul 5 by 11p;
ukinetes 15p in diameter; arthrospores 10.5 by 5p; oidiu 3 by 5.§p,
rare.

habitat; perusitic or semi-psrositic on cert in species of

,nrecu (pulms). ;/’

Thom, Co & Ii. Roper. 1949. :‘jlanI’itl

'9. 630—531. hilliums i hilkins Co.

l‘.‘
.L' l-)

2£_the Penicillin.

 

Colonies on tort geletine, producing salmon colored corenia
10 mm. in height or more; conidiophores obout 5p.in diameter;
metulue 7-l§p by 2.5-5.0p, irregulurly borne, irregulsr in number
or none; steriymute lO-BOp by 2.5-5.5p in groups of 2-5, or even 7;
conidiu ellipticel 5.0-7.Qu by S-gp.

.K-Lj‘i'i ’:
Gliocludium albumrfietch. 1937—33. Britisi TycolcgiCul éociety be: 251.

 

 

L ,
kin -' ’-
Fetch writes of e Gliocledium occurring on fiyxomycetes end,hed

been recorded as Gliocledium :enicillioides. a white crust is formed

 

over the sporungium, the miin stem being on in diameter with luterel //

brunches beginning et the first septum about EQp from the buse.
Brunches ere oyyosite or in whorls of’o ind divergent. Ihese brunches
sguin brunch and the conidioghores interlece. lhe phiulides are 13

by 1p. C:‘he stem end brunches ere minutely verrucose and lirger st

the nodes. The conidtu ore ovul or oblong-oVul, hyuline, 2.5-4 by

 

l‘SP” This is though by Ietch to be the same es Iencilliur elbum

-reuss und letch gives it the name Clio. album (Ereuss) Fetch.

 

 

 

Glioc1;dium strictun Fetch. 938. British Hypocreeles. Truns. Lrit.
‘ ' .- 1: r) o
-tc. soc. Hl.

 

Conidiel s+uge- conidioghores crowded, white in mess, hyuline,

100p hish, stem stout (4p diimeter be ow), fen brunches, lower bronches

olitury end distsn , upper brunches ghiulides opposite or in whorls
of three, ubout 30p. long, targering, :11 branches purullel to Inuin
stem. Conidie hyuline, oblong-ovdl or nurron oval, slightly in-
equuluteral, ends obtuse, 5-ll by 2-2.§p, the longer ultimately
.beccming one septute or pseudo-septete, united by mucous in h eds
about léu in dismeter, end in lerge :nsses.

Sexuel stuge;_§ygoxvces Brooxeenus Tul., Perithecis ovate,

 

_. O

.25-. mm in diuxeter, hyeline becoeing gole brown, densely clothed

in white or pele brown short tomentwd, excegt st agex, superficial
or immersed throurh overgrowth of the tomentum; esci cylindricil 150-
140 by 4p; sscospores fusoid, 1 septute, c0ursely sorted, shortly

1

spiculute st eecn end, hyuline, 15-16 by 3.5—gu.

 

Gliocludimn egesgitosum Petch. 1983-59. Gliocledium.Truns. Erit. Ive.

 

30C. L3: 357-263.

Conidioyhores forming write or yellow tufts, devuricute up to

.320.

J—‘

lSOP hifh. below U-dp in disneter. 4 both, brunched shove, the

1 wer brunches solitury, u1§er opgosite eech brun beuring u few
egicil 3h iliébs axich uie nrrro 1y uh.ulluceous Ihiulides 2gp hM

end 3F.dinneter. Conidis hfuline, oblong ov11 or re: only ov.1 some-

tines unieUelsterul, semetiues slightly elrved, smooth, 5-3 by i-gu.

On imxeture h ctriu mummoideu Plow.

 

GliOCludlld rgxnfcolu Jei. sp. nov. 1941. Sinensid 1;: Les. 1-6. 140-141.

 

"In frictibus Truni muse, Szechuun, Shine.

On rotten fruit, conidiophores upgeur es spurse downy growth
on the surfuce, erect, sin31e, occesionully brunched, septute, hyuline
or brown end guttulite at the bise, snooth, 133.3-302.4 by 7.7-12.lp
(exluding the penicillus); Le nicillus c;nsisting of one series of
metulue bedring e whorl of two to four sterigmotu with the extension
of the centrul uxis us the central aetulu, 25.3-44.3p high; metulae
cylindriCul, tapering to': 1rd the buse erd rounded ot the s; ex, 69.2-
67.2 by 2.5-6.Qp except the centrul metulee which mev be more then
lQu in diameter; sterignets 2-5, usually on e metulu, uciculur in
shege, rounded ut the buse und tupering torurd the tip, slightly
curved, tonerd the exis of the metulu, 11.2-16.3 by 1.4-2.1u;
conidia in chains then forming s heed with muc1lur110us fluid (43. 7-
123.§h in dismeter), elligticul to nurrorly ovste or elongited elliptic,
hyuline, biguttulute, very Virieble in size, 5.6-14 by 2.3-5.Qu.

Cn erur 1edium it yrodices a thin lug er of w ite down of fruit-
ing structure, luter snuff‘brOKn end the reverse cinnumon brown to
clove brown; uerisl rvcelium ut first none, tnen loose end 33erse;
vegetitive mycelium slender, 2.3-4.2p in diuueter end the fertile

uycelium much thiclcer and so eti es tinted with broun,b 6.0-9.1n in .

diuneter; CCDileChOIGS essenti 11y the Sure us thut on rotten fruits,
urisinr reruenciculir from the cells of ie tile hypliue, 67.2-333.4
by 6.3-13.9p; metulse of one to two series, or sometimes none, 11.1-
33.5 by 2.4-7.0u; sterigmutu forming u verticil on tie con idiophore
or in s group of two to severul on e metulu 7-lé by E-Sp; spores
1mi11r to those on the rotten fruits; sibmerzed :_ro nth consisting
of hylline mycelium nnd brown chlamydospores; chl “ydos sores ebund-
unt on old cultures, forming irregulhr bnlls of 4-13 cells, 4o.2_
61.2 by 6.9-5.2n; single cells globose to subglobose or irregul r
us the result of crov. ding, thick wulled, containing numerous lurge
oil globules, 12.6-29.4p across.

The Present species shons uffinity for G. roseum (Link?) Buinier
from thich it diffs: s by (l) the absence of loose floccose ty;e of
C~-rowth, (a) lurger conidia end (3) the ;resence of chlamydospores.

On fruit of Irunus muue it yrodices u rot cinn410n b ff in

 

color, soft, tith distinct and entire hnrgin, then covered with
whitish dorm ny neriul 510 th, moldy enell.distinct; infected flesh

honey yellow in color,"

Glioclsdiwla nirrum .Loresu. 1941. Rev. Lgcol. T. S. s-e: ol.

 

 

Under i’ortula und Levende bet.een Seinte- I: ergu its end
Formichet, francs.
Leriul mycelium Yhite, ebundunt on rich media (Molt 131r)-

only slichtly deve10"“d on DOOI‘ .uedis. ”ultires ere radiating: at
1 Ab - :

first co olorless 11d then with bluck zo sby the forhution of
conidiophores :1nd conidia \hich necroscopicullys @\e r in mess sooty
Ct

black. Conidioahores ty icul 01 liocludium severnl times brsnched
.L i O

the phiulides elongated more or less cylindricul ubout ldhclong.

L1 3
1‘0

F1

ihe conidia occ r in cLsins, cylindriCul, rounded st both ends,

oliVuceous, guttulhte. Length fighgfll, often Z/c, Uidth 2—2‘ und

excertionully 3/.. been little chains syglutinute in spheres lO-SQ/u
in dium; olivuceous-blcck which mqy flow together forxdng bluck
shining patches. This species reminds one of §:_viride Istruchct.
Eut is diStiASUiShSd by siy end form.of conidiu which ere bluck in
muss- nifrun indiCutes this lost cheructer. This gives to the
cultures the generul sSjLCt of the cultire of Lerutisceue but all

other cheructeristics nuke our fungus a true Gliocludium.

Glioclud um sollni (Ker ung) Fetch. 1945—45. “Cditionul notes on

 

British Hypocreules. Truns. ”rit. fiyc. Soc. 36-27: 149.

 

This is a trensfornstion and combination by Fetch of Iectria

soluni Reinke & Derth, as the sexuul stage and Spiclris Joluni

 

de Bury us the Eliocludium stige. . V”

The psrithecis sre described as being crowded on an erumgent
fleshy strogn; no definite cortex, forming pseudo-perenchymutous
tissue without sir spuces. Lomever when Schroeter grew it in culture
he found the hyphde did not fern pseudopdrenchymstous tissue. These
masses are soft, yellowish inte_iully, dark ed shove, and blsckish
externully.

The perithecis sre redlish or dull yellow end become elnost
white, elobose, .5-.4nn. in diameter with coniCul ostiolum. She
usci are nurrow cluvute, with truncute ugex, 65-78 by B-fip, the spores
are uniseriate or biseriute, uniseridte below. The ascospores are
OVul, narrow, hydline, beoihing somewhnt rough lO—lB by 4—Qp.
Conidiophores; scattered or greggriOus. Stems %H in disaster,

nooth, septnte, brunching beloa end bedring suberect luteral '

brunches. Conidiophores ere in whorls s tfe nodes end in clusters
st the u;e:, ull uttuining the some height.

rr

Conidid; ovul, 5-6 by S-gp, edge on end globose, hyuline

U

snooth remuining in e glohose or irreeulur Lies su‘?orted bf the

pniilices.

ocludiun cibotii (n.s.) Vun Beyms Thos hinged. lCéa-éS. ”ntonie Ven

 

 

 

 

M.
T A “ \F.» ‘P ‘N 4‘ 2 -. , ‘ “. "Y . ’, - r: " ' n ‘ ‘ "‘ r‘
n2erenhoek no rnil cl “icrooiolo : uze serolo-V. lu (l-~,: eo-es.
I. _
7., v-‘L ‘1‘ ' 4- ',~ "If .. I ‘ '. "<’ n 1" L‘"""" " W ‘
C.— ,' v’,‘,- —~“r- MU \A L- ‘U-Lr _. v.k .V J; J U ;\l .141 Dil.~\e .--O to: Ft Her
‘ ' e ralic e, t icr, ll *t coloro , .
I a - - —

in bundles m‘ich tren exhibit d green b oxn to brown bluck color.

Conidiojhores— to off iron the bundles of hyphue in RTEut

 

numbers, 30- CO by L-Qp,

I

Sterigfiutu- struigh , forming nwl-shupe, dC-4Qp long end the

 

buse an tide, mostly 5-3 verticillste.

Conidin- numerous, clusters glued together, hyuline, single
celled, ellitsoidol 4-6 by 2-2.7p.mostly 4-4.7 by 2.3-3.2p.with 2
oil droplets.

Hebitut- isolated from the huirs of the tree fern Cibotium

 

Scniedei Frl. L. F. Hubekotte, Delft.

 

. '\
Eure~afl~ure3

 

In Beer Fort uger- after one m nth, colony 7 cm. in
diumeter, end in the center severe unite bundles of hyghue 1 cm.
high. In the center s green-block felt like wrinkled met with
luxurient conidril formation. The edge consists of u zone of thin,

colored, sterile branches of hyphse 1 cm. high huving u delicute

(O
'1

nicy odor; the underside or the center is bluck, in other *leces

pole yellow. -

In test tubes to l4 dgys

 

EfiQE wort eesr- ut the bottom of the test tube, White

felt like met with sgirse conidiul development.

Cherry erer- ngsr surface covered with Shell numerous

 

thtish spore droplets. Here hyphsl bundles are raised, Bum. hieh
in cylindrical hyphul bundles which procuce numerous conidionhores.

Currot- covered with white felt like bundles of raised
hyph he .

?hole Iptuto- sine us cherry seer-,conidis nunerous.

 

Cut aeur- n shove except at the edge near the glues, e

——5——

blnck edge is Produced, in potato agar, absent.

lillitutie insienis (Uinter) n. on . Dennis erd Inkefield. 1345-46.

4 _\—(_

Yes or interesting fungi. Trsns. Brit. gyc. Soc. 39.

 

Un b9 Larch 1353 sgecine:S'of e fingus on mushroom.cungost from

Uxbridge, Yiddlesex, were received at Kew from.fir. J. Buddin. The
0

fru'tificetions were sghericul, glubrous, whitish to cream colored,
approximately 500p in dium. end scattered singly over the surface
of the congost. Licrotone sections showed them to have u stout
Wall, sons 80 to 90p.thick, canposed of ten to twelve layers of
cells, differ ntieted into an outer belt, three to four cells Wide,
of empty cells, and an inner zone, 7-3 cells wide, densely peeked
with oil globules. The center of the escocerp consisted of thin
Walled tissue in which were inbedded numerous esci, each with S
spherical yellow ascospores. The letter neusured ll-ldp in diameter
with e lsrge central oil globule end u well about by thick, densely
studded with short blunt Hurts. These measuregents tere mnde on

stores within usci. '

Excegt for store size, this agrees precisely with the dieghosis

of L. guillurdii described from humus in e glues house in angers,

 

Frufice, April lQCO. hccording to Loudier nnd Futouillerd (1900)
this fungus had spores zs—eep in diam. Type heteriul of the sgecies
was preserved by Est. end is now in the furlow gerburium. Er.
Linder re-exunined the msteriul und reported us follows. He found
the spores neusured 15.2-13.2 by 13.2-24.§p. Ihese spores were for
the most gurt still within the escus end were somewhut compressed
and engulur. In the few cuses were the spores hed been releused
from the escus, the spores are gerfect globose. It epgeurs there-
fore thut the spores of this fungus very rreutly in size and thet the
“users end Uxbridge collections represent the same species. It is
1 "
else the same us the furgus distributed under the heme g: Reggillioides

 

Cord; us hunter 150 of heliouqu/isr o innue. lhis hud been collected V/

v/

’F'

- 1". Po r“ “‘ .fi“ yr." /- ~ ‘ ~ . 1v vs ”2“va ? T)
on Cecljlflg seudeecs of ii‘tery -t. “dine, o. a. A., on,ojuui,!131o

In \ _.._— V
t r V’
by Thhxter, who cited us synonyms I. insione Juinier, Lihipenicilliuh

A , t/l/

insiene Brefelt end L. guillurdii Beud‘ end Put. (Thuxter, 135s).

 

 

Thom (1950), however regurded I. ifisifte us grobubly distinct from

G. penicillioides und the egplicuhility of the letter nlhe is ulso

 

dubious us will be s;own below.
C

Previously Winter (1375) hcd obtuined en geochy ete on goose
dung which Le regurded es the perfect stuge of G. :enicillicides.

 

Ho description mus published but anteriul with udeiuote figures of
l *i r 3 I ' V,
eeci and sscospores use distributed by Reb}nhorst;s fiungi Europuei,

w
(

no. 1733, under the nude Eurotium insigne G. Hinter n. e“. (en n. g.?).

‘v

 

Eruminltion of Eiiter’s heteri 1 st Yew shows it to correspond

closely with the nresent fungus exce;t thu the eSCOCurgs ere now u

&

m
C.

deco rellow brown end tie outermost cells s13eur deco *ellou in
a. 11 A A v

~-‘ ' rv " . -. >‘\ -- . a 1‘ ‘- c: ., 1 P\ V 'x \- .,
section. Que escosrores assaure lU-lquCI es. lessee end oulhcn

,1

(1331), “ho obtuined E. 'nsirne on duxg OI kunruroo, Cvis burrhel,

fowl, end horse, at Ket described it us huving the peritheciu cleur
p Y
white when young, then 117id, becoli_5 3ellonish std fin slly rusty

V,
bro n, with uscosgores globose, 17~aOCVucross. The color difference

mentioned uoove is thus clet1ly not si5nif icun nt end deiends probu:ily

A /
on the mode of greservetion. dinter's collection hos been dried for
/
1
70 years, the Uxbridge uuteriel hué been ke1t in form;1in.
These and other records of E. insifine huve been discussed by

Ietch (1959) who concluded thut the imperfect fungus ussociuted

with it is not G. nenicillioides which he ascribes to Evoomvces
— ’ M...—

 

§
I: 9/
eureo-nitens Tul. but G. mucrouodinum;3ur hul. The nlme aCOEted
a __ 4.. 4.

 

 

by Tl‘ exter is thus in p121ro Jriete both on the ground of mis-identi-

fic tion end of spplicubility only to the ingerfect stuge. ihough
r '
‘T

ussociution with u fliocludium nus not noted in the Uxbridge :zteri l

. I
Q
rpo.‘- J)‘ a

H 1

nor mevtioned by boud( u.d I:t., it dineu1s Lt present hishly probable

trut all the collections cited above are the edge species. owing to
S 1.

the vlry churucteristic end distinctive escospores it seems inudvis- 0/
I

able to refer this to Fungus to Eurotiun; it would eppeur preferbele
' V 7 ' ' ' ‘ 'I u .L . - . . It A L”
to retuin tn genus hilligutie, With the new combination L. 1nSistg.

 

J

 

. V"
M“ - . " ., .. . . v 1 .. , .
Bond. a nd Int, escrioed Lilliputiu to the Euberuceee, union lS
cleurly inedhissuble: Clarents and sheer (1931) trunsferred it to
r’

1

b . .. , . . 1 1
Gyhnoeseceee, in thicn it 18 cleurly out o: pluce. r

(D

:Cihg fietuiled
studies of the developing escocurp it seems to full most nuturully
in Eurotiiceue, nenr Eurotium, from which it is distinguished by

its sghericul united uses101es.

\

Gliocludiwn erreun finder. 1313. Ihytoput‘. be; 450.

 

Colonies on pototo dextrose ugsr or Czupek’s- at first pure
white liter yellow end occusionelly light slluon pink in fruiting
dress on Cznpek's medium,.mycelium loose-floccose, simple h3phue
or hyphue bunched in ropes; sclerotiu not observed; conidioghores
yer12ndiculor to ;edium, éC-lfifihn 1£nicilliu up to lep in length,
once or twice irregulurly brunched.

Phiulides lC-15 by B-Qp beering conidia which muy be eggreguted
in gelutinous messes or bulls (Acrostelefimus type) or agglutinited
in tygicul column (resanbling Clonostuchys type), conidia hyuline,
elliptical, epiculute, smooth, 3.3 by 5.9p extremes (;.6-3.6 by 5.1-
6.7g).

A slight to murked yellowing of the medium in I.D.4.

Hub. EuthOfenic on stored carrot roots in Y. Y.

To date on life history.

. . .. . . , 10 u . , t _
Optimum'lanp. el-eeOC- minimum below -o C. muXimun oetneen
as end 35°C.

(J)
(U

fT-r'v- Vrnv-w‘ ~m «1:1 ’1 ‘r-‘t.
v.1..LI. ‘ .

1.

4.

14.

15.

16.

17.

L-‘JAL-~& \(-L4—J V-__..¢—J

G. 1207.Glioc1;diun roseum. Lull. Soc. gycol. irunce.
23: lll‘liu, '1‘. 4;:1110 £10 :43]. Ii:_ 0 1‘5.

 

\ . .
beinler,.n. G. 1910. Lycotnegue ce 1' LCOlS he Inermncie. null.
Soc.2fiycol. xrirce. Lo: 3:5-530. T. KJVI.

Tutum. 1345. Ieurosroru II. um. Jr. Eot.

 

;e_C18, Go 1a,. b.1151 7‘. L
1 o 13

A
icidier, E. 1910. Icqnes KyoolociCue ou lconogru1hie Zes

Chn;;i5nons”2e Jrince. Tone IV Ilunche 517. see-547.

sou‘ie“, L. I. et Idtouillurd. L*OO. rote sur deux Chumsignons.
Bull. Soc. Tycol. firurce. lo: lee-146. T. XVI. Ilete V.

G..Lussee. 1337. 5046 exotic fun i. Greyillee

Ten British funni. 339. Grevilleu 17: 3C.

 

340. IJznor"ce.es. fees. Icones run:orum IV:
1. Tef. VII, fig. 93.

C)
O
*‘3
C11
1

0
I
I
o
(3
o
H
o
H

Dennis, R. A. 3., end B. K. Jukefield. lgéE—éd. Iew or interest:

r

ing fungi. Trees. Lrit. gycol. ro. L9: lee-147.

Dukes E. H 1943 The 12135 flo¢y of Dogestic -ni ‘ ls. X*'-7L1.
7v

0 0

5th edition. 3o took Publishing 30., 1thuCu, 1. 1.

eillflin, J. C. and 3. V. lhbott. Cot. lQLd-July 13L7. “ sumihry
of the soil fungi. loud gtmte College Jr. of ici. l

. Oct. lGLG-July 19L7. n suMLnry of ti~ _
Stute College Jr. of Lei. ‘1 (b): bUL, 303. fifi. 57.

. Cot. liao-Iuly 19L7. “ sunuiry of the soil fun _i. lone
Stute College Jr. of Sci. 1(3): 30;, 3Co.

1336. Ben r noteworthy fun i. Journsl of Lotdny :4:

Kerrick, J. n. 1942.1h13toouthologi0ul notes. Ihytoputh01053 DB:

1'1 ~'-‘
Dc.) 0 o

Lurchul, 3. 1395. Cnihpignons cogroghiles de 5 loicue. Dull. de

le Joc. Roy. de bot. de Lelg. 54: 125-199.

,—

flfl'c (tel 1h. -‘ 3*:
1:1

 

u.__
/ . . . . a

s1. Contributior:s u' l’ etude dos cnnmpiohons iruiticoes

de 'el {ficue . cull. soc. Ro' / ‘

9

‘. ;ot. Lela. oi: 10v.

 

1?. Iutrucnot, L. 1125. Chi:;i"r ore ccotyc tee. Lull. Leo. Col.
:jrarceo 3: ~/;Z-~E .40
1‘ “ore.u ““6 135-1 F7 1 L 5‘5rn (1 “:v ‘6
" . , “ . “ . ahb' —- . ~d—lOC-fi. ~ I - . Na L .0 . ‘b i . Ll
Lycoloji¢¢e ”-6' ol.

"3' ‘i .- qu- ‘ fl 5" ', '3 - H ‘ 1.»; 4- . w ‘I IV WY '5‘ ‘ -.. 3"“ - , T“ l . .
Lo. ietcn, 1. lose-L7. see1.1o.s to oe1lon -un 1. oe3lon U1. ~c1.

 

 

 

 

L . 1945-45. ndditionul notes on British fiy1ocreules. irons.
Brit. ILIVC. N"QC. iii-.147: 14:20
H.
2L. 13J3. Lrltlbh W pooreeles. irons. Lrit. “3c. -oc.
L .
£3. l~33~39. Glioclsdiun. Truns. Brit. ch. Soc. LL: 357- 563
“4. 11o“e1tcr, E. E. 1936 n cohgdretive stud3 of conidi-l foriution

in wequlOSyorluL end more relute ed h3phomyces. nnn. Ho.
Bot. Gurden Lo: 47-43.

35. Rabenhorst, L. 1337. LTvptOthenLlord von Ueutschlw d, Oesterreich
und der ochweiz. Lneite uufluge.1'(2): 61.

(H r'

26. Reder, H. E. IDéS. R izoctonie Carotue /. St. end Gliocl dime ,_

eureum n. 51., {fithew root utxo ens of our rots in

cold storere. Ihytopathology 53: 450.

 

 

 

 

L7. Succsrdo, I. n. 1367. Sylloge fun5orum hucusgue c05n itorum.
i: 30.

33. 1909. Totse I H'col 5iCue. Lnn'Lycol. 25 464.

29. 190C. 3311. Fun5orum.1§; 5L1.

30. ltlE. Syll. Auryorum.§3} 1379.

 

”P .1
51. Sopp, O. J. 0.1915. Z'ono 5ru‘iie der 3114e: r11 e Eenicillimn.
Tiucrsnccsclpgnriiu r I. Letnemu isK-neturvidensksbelig
klesse 1.91.23. l-cLCSo

32. Thom, C. 1360. The Ienicilliu. 644. villiQms 31d milking 30.,
Beltimore, 1d.

 

53. Vun beyne Thoe Iingme, don H. H. 1&24-45. Glioclsdium cibotii
fl“ 5‘ (fl. 3.). untonie Van Leetenhoek Journul of Licrobiology

and der0105y 10 (l-b): 40--5.

64. 1961. Uebeg\ein neues 31‘. oclsc u:1.Xitteilunjen use
den ”Cent rr\ cureuu voor ucninpelcultures". III.
Verhundelin5en der Koninklijke LKudemie vun fie
-etenscheygen te Lmsterdem. Tweede oectie. Leel

49. :o. 2.

 

 

Decetber 1941. Totes on the storgge aid mir:ot Ci
f fruits and varetgbles. I. Mgrket disegses of s
fniits. Sinensiu 13 (1/6 : 1J5-1SL.

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33. Uestergggrd, I. and H. I. Jitfihell. Dacaater lgéV.
1m. Jr. Bot. ”4: (10 : 575-577.

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37. “01f, 3. L. LiQ 2. 1. wolf. 1:17. TLe Lungi 2, Ajfidqo JOLn
In 110:" qud QOILS , Inc. , 1.67.5. 1. 0:1: ’ .- o :L’.

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