THE FUNCTIONS OF THE MICROSPORES OF THE HETEROTHALLIC
PYRENOMYCETE GELASINOSPORA CALOSPORA (MOUTON)
MOREAU ET MOREAU, VAR. AUTOSTEIRA
(ALEXOPOULOS ET SUN) SUN,
ALEXOPOULOS ET WILSON

by
Bernard

Joseph

Sloan

AN ABSTRACT
S u b m itted to the School for Advanced G raduate S t u d i e s of
M ic h ig a n S t a t e U n i v e r s i t y of A g r i c u l t u r e and A p p l i e d
Science in p a r t i a l f u l f i l l m e n t of the re q u ire m e n ts
for the degree of
DOCTOR OF PHILOSOPHY

Department

of

B o t a n y and P l a n t
Year

Approved

1957

Pathology

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Bernard

The
calospora

functions
var.

sented which
prim arily

of

autoste ira

as male

sexual

evidence

indicates

the

that

cells

microspores

are

demonstrates

Circum stantial

agative

the

cells
is

the

microspores

Evidence

microspores

(spermatia)

also

presented

is p r e ­

function

in f e r t i l i z a t i o n .
which

may f u n c t i o n

( m i c r o c o n id i a ) under

Sloan

of GeIasinospora

discussed.

that

Joseph

certain

as

strongly
asexual

prop­

environmental

cond i t i o n s .
Results
tization,

and

obtained

from

localized

perithecial

that

a trichogyne-microspore

this

fungus.

involving
indicate
are

not

somatic

a

Additional
study

that

is

associations

relationship

results

obtained

u-tube

and h y p h a l

compatible

nuclei

of

into

anastomoses
It

development,

the

brought

of

sperm atization, cross

close
and

suggested
only

in

associations

nuclear
that

the

this

is

sperma-

suggest

functional

in

from experim ents
tip

subcultures

h e t e r o t h a l 1ic
with

one

fungus

another

by

m igrations.

compatible

nuclei

a s c o g o n i a by way o f

occur
the

in c lo se

trIchogyne

SELECTED BIBLIOGRAPHY
1. A l e x o p o u l o s , C .
Ge l a s i n o s p o r a .

J . a n d S . H. S u n .
1950.
M y c o l o g i a ip2: 7 23 -73 1}..

A new

species

of

2.

S u n , H. S .
1953.
A c o m p a r a t i v e m o r p h o l o g i c a l and c y t o l o g i c a l s t u d y of G e la s i n o s p o r a c a l o s p o r a (Mouton) Moreau
e t M o r e a u , G e 1a s i n o s p o r a a u t o s t e i r a A l e x o p o u l o s e t Sun
and G e 1a s i n o s p o r a c e r e a l i s D o w d i n g .
P h . D. T h e s i s ,
M i c h i g a n S t a t e Co l i e g e .
5*5 p p .

3.

S u n , H. S . , C o n s t . J . A l e x o p o u l o s , a n d G. B. W i l s o n .
19 5 1 f A cytotaxonom ic stu dy of th re e sp e c ie s of Gelas i n o s p o r a .
C y t o l o g i a 19* 2 55-2 61 }..

1(_. T y l u t k i , E . E .
1955*
A s t u d y o f som e a s p e c t s o f m o r p h o l ­
ogy g e n e t i c s and c u l t u r a l b e h a v i o r o f t h e h e t e r o t h a l 1ic
p y r e n o m y c e t e Ge I a s i n o s p o r a c a l o s p o r a , v a r . a u t o s t e i r a
( A l e x o p o u l o s e t Sun) A l e x o p o u l o s and Sun.
P h . D. T h e s i s .
Michigan S tate U n iv e rsity .
73 p p .
5 . W i l s o n , G. B. a n d C. J . A l e x o p o u l o s .
1956.
Spontaneous
m u t a t i o n i n G e 1 a s i n o s p o r a c a 1o s p o r a , a h o m o t h a l l i c f u n g u s .
M y c o l o g i a lj.81 6 8 5 - 6 8 9 .

THE FUNCTIONS OF THE MICROSPORES OF THE HETEROTHALLIC
PYRENOMYCETE GEL AS INOSPORA CALOSPORA (MOUTON)
MOREAU ET MOREAU, VAR. AUTOSTE IRA
(ALEXOPOULOS ET SUN) SUN,
. ALEXOPOULOS ET WILSON

by
Bernard

Joseph Sloan

A THESI S
S u b m i tte d to th e School f o r A dvanced G ra d u a te S t u d i e s of
M i c h i g a n S t a t e U n i v e r s i t y o f A g r i c u l t u r e and A p p l i e d
Science in p a r t i a l f u l f i l l m e n t of the re q u ire m e n ts
for the degree of
DOCTOR OF PHILOSOPHY

Department

of Botany

and P l a n t

1957

Pathology

ACKNOWLEDGEMENTS

The w r i t e r
to

Professors

supervision
assistance
w riter
for
to

to

in

Philip

express
a n d C.

r e s e a r c h was

always

critical

its

G.

be

greatly

comments

B. W i l s o n

several

Coleman

for

the

long be
to

i i

whose

constant
Th e

both P ro fesso rs

guidance

photographic

under

remembered.

Appreciation

suggestions.

appreciation

and w hose

suggestions

his

stages.

sincere

Alexopoulos,

indebted

and

for

J.

his

initiated

and e n c o u r a g e m e n t w i l l

Professor

thesis

to

B. W i l s o n

this

w ill

their

G.

wishes

work

and

in p a r t i c u l a r

in o r g a n i z i n g
is

also

the

extended

and m any h e l p f u l

TABLE OF CONTENTS
Section
I.

Page

IN T R O D U C T IO N ....................................................

.............................................

1

...............................................................................

5

MATERIAL AND M E T H O D S ...............................................................................

9

Literature
II.

A.

Cultural

B.

Technique

C.
III.

Review

C o n d i t i o n s ....................................................................
Employed

in O b tain in g M icrospore

S u s p e n s i o n ............................................

10

1.

A p p a r a t u s ................................................ '.............................

10

2.

P r o c e d u r e ...............................................................................

12

3.

Dilution

........................................................

13

...................................................................

13

..............................................................

15

M i c r o s p o r e s ..........................................................................................

15

Technique

U-tube Experiments

RESULTS AND OBSERVATIONS
A.

9

I.
II.

Size

a n d M o r p h o l o g y ...................................................

Function
a.

Role

.....................................................
of M icrospores

. . . . . .

15
15

in F e r t i l i z a -

t i o n .........................................................................

16

............................

21

1.

Reciprocal

2.

Localized

cross

development

of

p e r i t h e e i a .......................................
3.

Spermatization
involving
of
i i i

22

experiments

definite

number

s p o r e s ........................................

23

iv
Section

Page
b.

Role

of Microspores

in Asexual

R e p r o d u c t i o n ............................................
...............................................................................

33

D IS C U S S IO N AND C O N C L U S IO N .................................................................

1+2

B.
IV.

27

M igrating

Function

M i c r o s p o r e s ........................................................

1+2

R e p r o d u c t i o n ..............................................................

1+2

R e p r o d u c t i o n ........................................................

1+6

SUMMARY....................................................................................................................

1+8

LITERATURE C I T E D .....................................................................................................

50

I.
II.
V.

of

Nuclei

the

Sexual
Asexual

LI S T OF TABLES
Table
I.

Page

Effect

of Microspore

Concentration

1 0 6 . 1 . ip o n P e r i t h e c i a l

1 1 3 . 1. 1
II.

Effect

Production

Strain

of

Strain

.....................................................................................................

of Microspore

21ip.l.l

of

Concentration

on P e r i t h e c i a l

of S tr a in

Development

of

Strain

2 1 i p . 1 . 3 .....................................................................................................

v

2 ip

25

LI ST OF TEXT FIGURES
F igur e
1.

Page

Filtering

Apparatus

Suspension
2.

Various

Stages

Used

to O btain M y c e l ia l- f r e e

of M icrospores
of Microspore

.............................................................
Germination

11

on S o l i d

A g a r ............................................................................................................................. 3 2

vi

LI S T OF PLATES
Plate
I.

Page
Sexual

structures

a u to ste ir a
II.

Perithecial
involving

of Gelas inospora

..................................................................................................................ip
formation

resulting

microspores

p rotoper i thee i a of
rocal

of

strain

spermat i zat ion

phoditic

calospora v a r .

compatible

from

strain

211p.l.

21ip.l.3.

involving
strains

sperm atization

the

1 and

Also

recip­

t wo h e r m a -

21 Lp - 1 - 3 a n d

2 2 7 . 3 . 7 .................................................................................................................... 2 0
III.

Results

of

som e o f

experiments

the

u-tube

and h y p h a l

...........................................................

tip

. . . . . . . .

37

SECTION I
INTRODUCTION
G e 1a s i n o 3p o r a c a l o s p o r a v a r .
by Sun,

Alexopoulos

found

to

1955,

Tylutki

and W i l s o n

be h e t e r o t h a l 1 ic
(35)

demonstrated

hermaphroditic

com patibility

groups

spermatia

author

also

by most

of

this

spermatial
been

Since

had

there

the

fungus,
and

done
been

to
no

auto ste i r a ,

conducted

to

structures

and

With

aims

these

the

able

the

relationship.

produced

i t was

suggested
possible

of

(9)

r e s e a r c h was

mining whether:
1

certain
existence

in

1950

adJuneta

structures
to

but

function.
the

function

b y G e 1a s i n o s p o r a c a l o that

an

function

investigation
of

m e c h a n i s m w h e r e b y plasmogam y was

in mind,

spermatia

possible

in r e l a t i o n

both

latter

under

in G e la s ino s p o r a

bodies

of

produce

The

Cain

In

f u n g u s was

to

formation

indicated

the

(1).

ascospores

particularly

s t u d y made

originally

this

in c u l t u r e .

show how t h e s e

determine

that

single

the

described

and Sun

equally

time

production

s p e r m a t i u m - 1 ike

spora var.
be

this

conditions,

no w o r k h a s

of

of

at

a tric h o g y n e - spermatium

reported

are

and p r o t o p e r i t h e c i a

isolates

environmental

further

since

normally

described

1 9 5U > w a s

in

by A lexop oulos

fundamentally

both

(33)

a u t o s t e ir a as

directed

these
initiated.

toward

deter­

2
(1)

the

microspores

functioned

as

sexual

male

microspores

functioned

as

asexual

cells

( spermat i a ) ,
(2)
of

the

reproducing
(3)

sexual

the

the

organism

microspores

whether

(m icronconidia),
functioned

r e s e a r c h was

sperm atization

is

the

h e t e r o t h a 1 1 ic

expressed

opinion

that

the

the

past

numerous
becom e

tend

other

to

been

w ays

asexual

the

sperm atization,
plished,
between

would

only

and

seem to

of

through

be

through

the

to

of

Plate

I),

opposite

hyphae.

of

other

or

be

to
has

species

related

could

the

able

described

somatogamy,
hyphae

are

production

available

possibilities,

initiated

(see

nuclei

somatic

in

has

spermatia

ignore

fungi

several

methods

and v e g e t a t i v e

these

and

cross

(13)

discovered

importance

situation

genus
than
accom­

copulations

compatible

additional

lines

strains.
of

determine whether:

a trichogyne-m icrospore

fungus
(2)

possible

Dodge

macroconidial
the

determing

insuring

fungus.

whereby c r o s s - f e r t i l i z a t i o n

i n v e s t i g a t i o n were
(1)

any of
the

of

h e t e r o t h a 11ic

Since

unlike

tr ichogynes
Because

this

for

toward

who h a v e

their

in which

reported

G elasinospora,

those

magnify

cross-fertilized.

Neurospora,

in

both

o n ly method

in t h i s

of

as

directed

fertilization

never

capable

structures.
Additional

in

cells

relationship

was

existent

or
mating

ty p e s were

transferred

3

PLATE I

F i g u r e s A.

C

Sexual
var.

F i g u r 0 A.

structures

o f G e 1a s i n o s p o r a

calospora

autostar i a .

Young p r o t o p e r i t h a i c a 1 f u n d a m e n t w i t h
singla

unbrancad

trichogyne
ca.

F i gure B .

Several
phores.
the

dichotomously branched
The m i c r o s p o r e s

individual

its

cells

of

are
the

x lplj.0

microsporo-

produced

from

microsporophores
ca.

Figure

C.

A single

germinating

spore w all
long

still

microspore.

remains

attached

x

1000

The m i c r o to

the

germ t u b e .
ca.

x

1000

5
Literature

tion

Since

the

the

fungi

in

Olive

(30),

discussed

literature

only

in

concerning

general

those

R e v i ew

cases

have

been

pertinent

methods

of

discussed
to

the

fertiliza­
recently

by

problem w ill

be

here.

Microspores
are

known t o

the

algae

be

(spermatia,

found

p y c n i o s p o r e s , or m i c r o c o n i d i a )

characteristically

(Fiorideae),

fungi

(rusts,

in c e r t a i n

a s c o m y c e te s ),

groups
and

of

in

1ich en s.
D efinite
function
until
in

the

1850,

and

as

early

that

Tulasne,

agents

small
these

small

bodies

later,

1861

would

to

function

Stahl,
Tulasne.
of

the

He

according
actually

lichen

fungi

fusing

with

cribed

h ow t h e

nated
have

in h is

that

tips

of

his

like

in

to

Backus

Backus

(7),

b y som e

may p r o v e

to

Itzigsohn

be m a l e

the word
small

literature

ascolichens,

to Drayton

coined

(6),

the

even went

ele­

(19),

in

spermatium,

structures

further

process whereby

fertilization

of

the

pollen.

Humphrey

mycelia.

(23)

in

than

spermatia

by a t t a c h i n g

Sclerot inia

and p r o d u c e d

work.

appear

so m e d a y t h e s e

trichogynes.

microconidia

a s c o m y c e t e s ma y

according

described

effect

cultures

questioned

to

in the

produced

structures,

in

show n

not

structures

a year

these

did

According

and p r o p h e s i e d
be

microspores

thirties.

discovered

describing

that

fertilizing

indicated

ments.

proof

and

1893

fr uet igena
However,

des­

germi­
m any

6
In
that

in most

depends
and

the

L a b o u lb e n ia le s , Thaxter
species

upon

the

subsequent

However,

described,

direct

passage

cytologicai

of n u c le i
to

has

development

attachment

data

(3 l|)

of

of

the

spermatia

dow n

confirm

suggested

into

the

these

ascocarps

to

trichogynes

ascogonia.

findings

are

1a c k i n g .
Higgins

(21,

e 11 a d e s c r i b e d
and

even

the

author

the

22)

formation

illustrated

spermatia

was

Backus

in

h i ema1i s , d e s c rib e d
chogynes
and

the

but

actually

as

ability

numerous

m icroconidial

and

imperfects,

attached

to

any d i r e c t

in p e r i t h e c i a l

fusions
ever

indicated

descriptions

(19),

Dodge

evidence

that

but
the

development.
the

the

of

in

up t o

to

fungus

Coccomyce s

themselves
spermatial

remarks,

to

tr i­

bodies

reports

dem on­

fertilization.

types

produced

the

no

1930 w h i c h

effect

various

reportedly

appeared

spermogonia

trichogynes,

previous

literature

had

of Mycosphar-

seen.

of m ic ro s p o r e s

bodies

and

attach

between

from the

the

species

trichogynes

how m i c r o c o n i d i a

in

two

19 3l_j_, w o r k i n g w i t h

actual

appeared

the

However,
or

no

find

tric h o g y n e s were
Thus

strate

to

any p a r t
(6)

of

spermatia

unable

play

working w ith

in

of
the

literature

spermatial
ascomycetes

prior

to

the

year

this

date.
Drayton
demonstrated
spora
the

conclusively

sitophila,

microspores

(11)
that

Neurospora
act

in

a n d A m es

(3)

in S c l e r o t i n i a

tetrasperm a,

a fertilizing

in

glad i o 1i , Neuro-

and P l e u r a g e

capacity.

1932,

anser in a ,

spores

in N e u ro sp o ra

sps.,

w ill

effect

germinate

not

and

give

protoperitheeia
gether

with

observed

in

which

has

germinate

dia

dung

is

(38)

to

be

the

only

year

1937,

a colony but
In
species

by Bombard i a

incapable

be

solely

G e I a s i n o s p o r a , G.

as

authors'

description,

spora,

a cytological

a few y e a r s

proposed

that

the

to

the

later,

situation

anserina

as

of

not

from

a new s p e c i e s ,

B om bar-

(39)

how

described

produced

in

laterally

flask
on

and S c l e r o t in ia
of

(1)

germination

described

the

Alexopoulos

like

glad i o 1i ,
to

form

agents.
a new
found

cross-fertile.

of

the

spermatia,

s p e r m a tia were

n am e G e l a s i n o s p o r a

do

ascomycete

These

investigation
Sun,

in

ascocarps.

a u t o s t e i r a , which was

s e 1f - s t e r i l e , b u t

to­

ascocarps.

fertilizing

and Sun

produce

be m at ed

formation

1u n a t a .

be h e t e r o t h a l 1 i c ,

Following

can

also

microspores

developed

show n t o

original

turn

micro-

g am et es w h i c h when p l a c e d

Zickler

anserina

19^0, A le x o p o u l o s

of

it

by P le u ra g e

functioned

the

s p e r m a tia were

produced

sim ilarly

the

described

hyphae

were

in

a coprophilous

sperm ogonia which were

produced

that

the

but w ill

produce

contrasted

about

shaped

those

to

as male

isolated

uninucleate

These

that

g 1a d i o 1 i a n d P l e u r a g e

bring

1 9 3lp a n d
In

strain

demonstrated

bodies

in

1u n a t a .

colourless

this

function

Zickler

f e r t i 1ization

and m i c r o c o n i d i a .

been

but

demonstrated

to m ycelia which

Sclerot inia

on r e c e p t i v e

horse

rise

a compatible

However,

it

only

Dodge

not

In the
reported.

genus

Gelasino-

and W i l s o n

calospora

to

var.

(33)

autoste ira

8
be

used

the
form

rather

than

h e t e r o t h a 11 ic
( G.

form

investigation

while
of

(35)

produce

spermatia

reported

was

investigated,

Of

the

the

as

f unct io n .

though

relation

has

described

spermatia

ditions,

function

remaining

production

produce

name,
the

a morphological

certain

been

of

of

the

to

separate

homothallic

of

and

genetical

autosteir a ,
this

Although

fungus
an e x c e l ­

s p e r m a t i a was p r e s e n t e d

s p e r m a t i u m - 1ike
suggested

that

bodies
a trich-

exist.

species

reported

by C a i n

the

i t was

does

six

strains

in c u l t u r e .

description

author,

ogyne-spermatium

that

regularly

morphological

juncta.

auto s te i r a ) from

conducting

by t h i s

tial

( G.

described

G e 1a s i n o s p o r a c a l o s p o r a v a r .

Tylutki

not

original

calospora) .

Later,

lent

the

(9).

in

this

fungus

no w o r k w a s

done

to

of G e la s in o s p o ra ,

only

i n G e 1a s i n o s p o r a

A l t h o u g h C a i n was
under

sperma­

certain

s h o w how t h e s e

able

cultural

structures

adto
con­

SECTION I I
MATERIALS AND METHODS
A. Cultural
A description
strains
method

employed
used

reported
as

the

this

original

study were

in m a i n t a i n i n g

( 3tp) .

outlined

in

of

stock

Furthermore

by T y l u t k i

cultures

of

the

cultural

conditions

Conditions

the

(35)

isolates
obtained,

cultures
standard

were

has

as

the

as w e l l

the

employed

as

been p re v io u sly

cultural

protoperitheeia 1 strain.
are

from which

here

conditions

in

These

growing

standard

follows:

( 1 ) Med i um:
Difco

c o rn meal

agar,

17 g m s .

in

a

liter

of

agar .
(2)

Temperature:
T w e n ty d e g r e e
temperature

(3)

C,

obtained

control

by

using

a B.O.D.

cabinet.

Light:
A Sylvania

ll^ -w a tt c o o l

cent

with

tube,

the

ballast

placed

outside

placed

approximately

1e v e 1.

the

light

unit

incubator.
2 8 cm.

standard

removed

The

from

flores-

tube

the

and
was

culture

10

The m i c r o s p o r e - p r o d u c i n g
tubes
tube

containing
corn

meal

incubator
time

and

20 m l.

agar

strains
of Difco

slant

incubated

were

in

co rn meal

cultures

for

grown

were

large

agar.

placed

approximately

ten

were

test

These

inside
days

sufficient

quantity

of m ic r o s p o r e s

B. Technique

Employed

in O b ta in i n g M ic ro s p o re

test

a 2 8 °C

at

which

produced.
Suspension

1. A p p a r a t u s
The m o s t

satisfactory

suspension

of

the

ments,

was

to

filter

layers

of

used

is

filter

as

microspores,

into

side
cm.

the

neck
test

the
No.

Then

glass

microspore

rubber

of

viable

of

an a q u e o u s

m ycelial

suspension

tube which

filter
base

of

the

paper
the

frag­

through
filter

over

a p p a r a t u s was

first

fifteen
apparatus

the

had

several

apparatus

sterilized
pounds
is

or

placed
and

cotton

entire

fitted

this

was

flask

been p re v io u sly
Four

were

a microspore

and

was

E r le n m ey er

funnel

was p l a c e d

funnel

stopper,

flask.

wrapped w ith

In p r e p a r i n g

a glass

a 5>00 m l .

Erlenmeyer
I

the

of

of

f la s k were

filtering

obtaining

A description

tube

9 one-hole

large

at

free

of

foilow s:

a No.

minutes

the

paper.

The

a

method

five

neck
and

inserted

containing
placed

layers

inside

the

region

with

of

of

the

apparatus.

in

pressure

illustrated

the

an a u t o c l a v e
per

square

in t e x t

filtering

for

thirty

inch.

figure

The
1.

12

funnel.

a 14.0 0 - m l . b e a k e r

suspension,

in­

11

Figure

1.

F i l t e r i n g A p p a r a t u s Used t o O b t a i n M y c e l i a l - f r e e
Suspension of Microspores.

12
2.
All

strains

employed

grown

in

test

bated

at

28°C (±°C) for

spore

production

microscope,
tube

tubes

on

for

slants

of

Difco

discernible

distilled

until

microspore

approximately

became

sterile

cultures

Procedure

the

entire

co rn meal
ten

with

water

was

agar

p r o d u c t io n were
agar,

days.

Wh e n m i c r o ­

a binocular
pipetted

slants

incu­

dissecting

into

became

the

test

completely

inundated.
The

test

palms

of

both

fifty

times.

effective
phores

hands

cultures
and

to

growth

covering

minimized

the

The
t u b e was
and

the

of

then placed

back

and

microspores

in
the

agar

amount

obtained with

microspore

several

collected

inside

sterile

test

the

inserted

test

suspension

each
by

tubes.
on

aid

of

sperm atization.

any of

the

microspore

the

be

quite

microsporothe

somatic

Therefore,
viable

this

mycelial

microspore

check
to

No h y p h a l
suspension

of

suspension

see

each

forceps

filter

paper

Sterile
into

the

of

the

if

hyphae were

the
plugs
mouth

microspores

fragm ents were
samples

test

located w ithin

apparatus.

sterile

w e r e made

before

to

degree

layers

tubes

filtering

Microscopic

slides

approximately

suspension w ithin

through

of

the

the

procedure.

poured

flask

from

of

between

found

slants.

then

then

forth

any s e r i o u s

actual

filtration

aqueous

Erlenmeyer
were

the

disrupting

f r a g m e n t s w h i c h ma y b e
prior

rolled

in d i s l o d g i n g

procedure

were

T h i s m i l d m a n i p u l a t i o n was

without

mycelial

tube

tested.

present

detected
The

in

in

13
microspore

suspensions

were

3.
In order
spores,
counts
ber.

to

microspore
w e r e made

This

squares,

each

Dilution

spermatize

is

of

is

each

the

exact

center

involved

an

chamber

the

a total

subdivided
eighty

of

of

into

count,
and

cedure,

a series

a total

the

into

the

divisions
trial

making

the

of

for

divisions.
square
in

use.

a grand

spore

This
further
of

made

until

count

Following

dilutions

of

the

total

counts were

suspension.

cham­

twenty-five

located

actual

of

counting

o f w h ich was

spore

Then

of m icrospore

use.

and m i c r o s p o r e

sixteen

square

each

squares

each microspore

for

number

outermost

five

a v e r a g e was m a i n t a i n e d .
for

into

selected

Repeated

derived

prepared

chamber were

sixteen

squares.

a definite

subdivided

a microspore

of

ready

Technique

with

divided

of which

and

a Howard C o r p u s c l e

In d e t e r m i n i n g
corner

labeled

su sp en sio n s were

by use

chamber

then

this

was
pro­

was p r e p a r e d .

C . U-Tube E x p e r i m e n t s
U-tubes
mine w hether
opposite
var.

used

nuclear

mating

in

this

m igration

types,

could

study

in

and m y c e l i a

occur

an

attempt

to

dikaryotic

i n G e 1a s i n o s p o r a

deter­

for

calospora

au to s te i r a .
The

three

were

hours

previously

u-tubes
at

were

160°C

prepared

and
and

heated
then
held

in

an oven

c o r n meal
in

test

for

approximately

agar which

tubes

for

had been

a few d a y s ,

1U
was

remelted

ify.

and p o u r e d

Following

blocks

of

agar

inoculation
containing

the m ic ro s p o re s,
sicator
This

placed
quite
in

were

as

inside

still

found

of

were

be

after
free

arms

and
of

or w i t h
placed

distilled

The m o i s t

drying

of

months

of

of

spaces.

air

the

to

u-tubes

in

a

the

chamber

large

was

agar

incubation,

of
des-

bottom.

procedure
the

solid­
with

a suspension

This

out

allowed

inside

water

incubator.

in p r e v e n t i n g

to

the

chamber.

(+_1°C)

Even

u-tubes

myce1i a ,

sterile

a 20°C

u-tubes.

the

u-tubes

a moist

effective

the

the

containing

served

into

the

then
was

medium
u-tubes

SECTION I I I
RESULTS AND OBSERVATI ONS
A . Microspores
I.
The
bodies

as

au to ste ir a
Tylutki

( G.

(35)*

and

of

the

by many s t r a i n s
a u t o s te ir-a) have
It

was

also

trichogyne-spermatium

fungus.

In

mentioned

phology

the

In

of

addition,

authors

s p e rm a tiu m - 1ike

however,

facts

s p e r m a t i u m - 1 ike

bodies

tions

through

and

occurring

are

in

and

of

been r e c e n tly
in

the

that

protoperithe-

relation

to

have
the

possibility
hyphae

described
report

exists

results

concerning

somatic

the

spermatiophores

of Gelas inospor a c a lo s p o ra

bodies,

the

s p e r m a t iu m - 1ike

branched

relationship

investigation,

previously

of

formation

indicated

possible

this

formation

and d i c h o t o m o u s l y

a description

fundaments

and M o rp h o lo g y

morphology

on u n b r a n c h e d

as w e ll
cial

size,

Size

by

that
in

a

this

obtained

by

the

and m o r ­

size

the

been v e r i f i e d .
function
of

have

of

nuclear

been

the
m igra­

investigated

presented.
I I . Function
An a t t e m p t w a s m a d e

tium -like

bodies,

microspores,

to

determine

which h e n c e fo rth

shall

o f G e 1a s i n o s p o r a c a l o s p o r a
15

var.

whether
be
var.

the

sperma-

designated
autoste ir a

as
( G.

16
a u to s te Ira ) function
cells
the
to

sex

cells

( m i c r o c o n i d i a ) endowed w i t h

organism,
produce

while

or

as

both.

Since

only p r o to p e rith e e ia

others

or w e a k ly
both

a s ma l e ,

have

fertile

mating

types

been

found

the

capacity

to

s o me

isolates

have

or m i c r o s p o r e s

to

be

in com patible
designated

(sperm atia),

completely

matings,

as A and B were

asexual

reproduce
been

found

in c u l t u r e
cross

only

as

t wo

sterile

strains

selected

of

for

study:
(1)
strain,

a pr otoper i thee i a 1, m icrospore

"A”

producing

" B"

"211+. 1 . 3 , "
(2)

strain,

a p r o t o p e r i t h e e i a 1,

microspore

” 2 2 7 . 3 . 7 , ,f
(3)

and

TfB ” s t r a i n s ,
The
these

producing

two n o n - p r o t o p e r i t h e e i a l , m i c r o s p o r e

” 2 lip. 1 . 1 , "

description

isolates

were

employed

for

each

of

reported

by T y lu tk i

and
of

” 106.1.1+.”

the

obtained
the

four

( 35>) .

producing

It

original
as w e l l

strains

from which

as

the

designation

isolates,

has

been p r e v i o u s l y

is

as

system

follows:

E a c h s e g r e g a n t was g i v e n a number in t h r e e s e c t i o n s ,
000.00.0.
The f i r s t s e c t i o n [ 0 0 0 . ] o f t h e n u m b e r was
t h e nu mb e r o f t h e p e r i t h e c i u m f r o m w h i c h t h e a s c u s was
t a k e n ; t h e s e c o n d s e c t i o n [ . 0 0 . ] was t h e number of t h e
a s c u s , and t h e t h i r d s e c t i o n [ . 0 ] was t h e s e q u e n t i a l
number g i v e n t o t h e s p o r e as i t was removed in t h e
s e r i a l d i s s e c t i o n of the ascus.
a # Role

of Microspores
In

an a t t e m p t

microspores

to

in F e r t i l i z a t i o n
determine

of G elasinospora

the

calospora

possible
v ar.

funtion

autoste ira

of
( G.

the

17
autoste i r a )
m e n t s was

dish
a

t wo p e r i t h e c i a l ,

separately
cultures.

laboratory

imately

on 20 m l .
These

incubator

in

test

tubes

cultures

were

incubated

found

tion

to

favor

to

the

tube

test

tion

over

at

spores

at

slants.

this

were

was p r e p a r e d

hyphal

for

sperm atization.

unaided
over

the

0 . 2 ml.

experi­

eye,

four

areas
of

incubated
at

this

No

such

time

were

an

agar,
sterile

where
aqueous

to

glass

using

However,

ten
the

days.
forma­

upper

found
of

had

the

micro­
check

suspension

ever

detected.
plate

discernible
placed

on

of

cul­

with
the

been produced,

of m icrospores

end

scattered

the

on t h e

were

hours

microspores,

the

also

tube

forma­

A microscopic

easily

rings

test

The

least

at

were

temperature

revealed

produced

protoperitheeia
suspension

the

at

fra g m e n ts were

and w e r e

approx­

twenty-four

suspension

above.

were

These

higher

of

abundantly

described

for

temperature.

for

inside

strains

formation.

quantity

f r a g m e n t s w a s made p r i o r

c o r n meal

this

were

in P e t r i

20°C(_rl°C)

slants.

since

however,

Wh e n p r o t o p e r i t h e e i a
of

agar

An a q u e o u s

as

agar

illum ination

particular

quite

Some,

slant.

at

occurred w ithin

cultures

for

tures

kept

constant

a sufficient

the

entire

were

28°C

usually

cultures

of

agar

the

of

strains

two m i c r o s p o r e - p r o d u c i n g

of m icrosporophores

the

series

c o r n meal

maxi mum m i c r o s p o r e

obtain

Observation

Difco

on c o r n meal

incubation

in o r d e r

of

The

of m icrospores

following

of

under

days.

was

following

microspore-producing

cultures

eight

grown

the

employed.

The
grown

in f e r t i l i z a t i o n ,

the

the
agar

and

18
opposite
The

mating

remaining

tilled
order

water
to

of

were

small

areas

the

microspores

perithecia

No p e r i t h e c i a
ceived

drops

set

repeated

up

the

inside

in

the

of

the

ml.

in

of

the

All

20°C

asci

ever

times.

rings.

sterile

dis­

carryover

suspension,

twenty-four

with

Plate

mature
in

distilled
of

II.

water.

twenty

In ev ery

under

con­

hours

produced

of water

pitted

rings
This

or more
instance,

over

carrying

Wh e n m a t u r e ,

dark,

those

t wo

plates

thirty-six

drops

of

several

and k e p t
to

In

or

in P e t r i

young p e r i t h e c i a w ere
rin g s where

one

over

cultures

incubator

produced

replicates

glass

experiment.

possible

microspore

been p laced ,

sterile

numerous

these

ascospores.

which

had

re­

experiment

has

Petri

plates

identical

and

results

obtained.
The

one

of

against

plates.

treatm ent,

were

0.2

a control

W ithin

contained

t wo

s u s p e n s i o n was p o u r e d

to

had

in

received

as

the

agar

illum ination.

the

were

in

returned

this

placed

a control

microspore

following

been

rings

served

c o r n meal

then

stant

and

fragments

the

sterile

glass

m aintain

m ycelial
ml.

type were

or

several

t wo m l .

of

co rn meal
the

agar

microspore

plates

which

suspension,

had

received

remained

sterile

ind i c a t i n g :
(1)
micro spore
(2)
vegetative
the

lack

of

any v ia b le

mycelial

fragments

in

the

germinate

and p r o d u c e

suspension,
inability

of m icrospores

m y c e l i u m on D i f c o

environmental

conditions

to

corn meal
employed.

agar

at

least

under

19

PLATE I I

F i g u r e s A-

Perithecial

formation

of microspores
thecia

F i g u r e A.

of

Perithecia

the

of

produced

with

21i|.l.l.

in

control

F i gur e B .

Sa me

as

F i g ur e C .

Reciprocal
strains

F i gur e D.

protoperi-

type.

resulting

of p r o to p e r i t h e e ia

of

from the

strain

of m icrospores

Perithecia

failed

to

of

develop

rings.

cross

in which

between
the

placed

t wo

hermaphroditic

microspores
on t h e

of

strain

p r o t o p e r i th ee i a of

227.3.7.

Reciprocal

cross

strains

in which

227.3.7

were

of

rings

over

placing

i n F i g u r e A.

2l L| _. 1. 3 w e r e
strain

in

from the

type

mating

a suspension

strain
the

one m a t i n g

opposite

sperm atization
2114- . 1 . 3

resulting

strain

between
the

placed

211j..l,3.

two h e r m a p h r o d i t i c

microspores
on

the

of

strain

protoperitheeia

20

21
1.

Reciprocal

zation

cross

Since

it was

cross

between

hermaphroditic
the

the

microspores

were

of

the

placed,

Plate

mat i z a t ion
never

in

drops

experiments

on
just

germ t u b e s .
capable
under

of

peared
var.

were

In

areas

relation

it

to

placed

several

even

detected

However,

at

reproducing

growth

of

this

certain

that

male

designated

the

the

( G.

the

hours

the
that

the

been

time were

these

of

the

The m i c r o ­

long

on a g a r
slender

germ t u b e s

after

apparently

sper­

microspores

volume

extremely

organism even

had

reciprocal

agar.

normal

rings

described

suspension

c o r n meal

proto­

the

microspores

as

and

perithecia

by

previously

their

a rtifi­

227.3.7

delineated

aqueous

in which

several

found
weeks

favorable

for

organism.
obtained

microspores

at

this

cells

spermatia.

and

time,

o f G e 1a s i n o s p o r a

autoste i r a ) functioned

propagative
as

fertilize

producing

results

the

to

the

c o n d i t i o n s w hich were

of

autosteira

no

strain

was n o t e d

times

to

of

as w e ll

on D i f c o

conducted

employed

the

s p e r m a t i>

cross-fertile

thirty-six

containing

described,

In view

sexual

Within

immediate

of water

to

cultural

mycelial

strain

m y c e l i a when an

swelled

s o me w e r e

latter

sperm atization

m i c r o s p o r e s was

and

2 lit. 1.3 w e r e

former.
the

a reciprocal

an e x p e r i m e n t was

protoperitheeia

the

II.

produced

spores

of

t o make

s e 1f - s t e r i l e , b u t

strain

the

the

produced

where

of

of

fertilize

per ith e c ia

t wo

strains,

microspores

cially

desirable

only

therefore,

Furthermore,

the

it

calospora

in th e
the

ap­

capacity

microspores

localized

22
perithecial
the

development

appropriate

cial

areas

of

mating

the

within

type were

mycelium

tends

ogyne- spermatium r e l a tio n s h i p
2.

Localized
The

tained

in v a r i o u s

previously
t wo

development

localized

(0.2

ways,

be

strain

where

no p r o t o p e r i t h e e i a

of

resulted

the

the

glass

spores

of

c o r n meal
plates
the

in

sides

of

after

the

in

these

mating

type

development

and

the

inoculated with

containing
strain

that

of

method
results

of

on y o u n g c u l ­

As
the

soon

but

as p r o -

placing

these

of

structures

perithecia

inside

on one

of

side

sides

of

sterile
later,

occurred

microspores,

21 Ip - 1 . 3 h a d

sterile

the

2 lip - 1 . 3 o n e w e e k

of p e r i t h e c i a
the

grown o u t

when m i c r o ­

p r o t o p e r ith e c ia

the m icrospores

on

of

experiment,

opposite

strain

development

and p r o d u c e d

been ob­

suspension

rings

areas,

placed

mycelium of

the

developed.

2 lip. 1 . 1 w e r e

plates

has

m ycelium had

strain

the

trich-

fungus.

from the

glass

sim ilar

microspores
suggests

localized

localized

to

microspore

the

had y e t

opposite

plates,

a possible

this

addition

In a n o t h e r

agar

in

protoperithe-

of p e r i t h e c i a

rings.

were

s a me

inside

produced

the

suggest

evident

the

21 Lp. 1 . 3 w h e r e

to p e r ith e c ia were
microspores

in

is

of

2 1 1 p . l . l was p l a c e d
of

to

on t h e

of

described.

ml.)

tures

placed

when m i c r o s p o r e s

of p e r i t h e c i a .

This

now t o

Droplets
strain

other

rings

existent

development

described.

experiments

the

but

grown o v er

on t h e

only
the

in

the

areas.

remain v ia b le

and

that

This

they

23
function
sexual
3.

as

sexual

structures

fertilizing

agents

as

soon

as

the

female

develop.

Spermatization

experiments

involving

definite

number

of

spore s ,
In
offer

an a t t e m p t

to

obtain

additional

further

evidence

of

a possible

relationship,

a series

of

experiments

over

sixty-five

strain
mate
be

113.1.1.

number

placed

type,

In

this

glass

determined.

incubated

for

illum ination.

2Ilp.l.I,

and

slants

suspensions
use

of

both

used

to

experiments,
Two

groups

sterile

of

rings.

With

set

up

for

the

the

one

each

eight

the

the

of

days

in t e s t

28°C f o r

at

1 0 6 . 1 . ip t o
opposite

counting
two-fold

series

tube

II.

glass

rings

were

and

mating

0 . Ip m l .

of

microspore

of

four

dilutions

in

technique,

on t h e

the

ma d e b y

dilutions

agar

microspore

ty p e was p l a c e d

replicas

Microspore

described

dilution

placed

under

on c o r n meal

counts

as

of m ic ro s p o re

2 lip - 1 - 3

strains,

days.

chamber

mating

20°C(+°C)

tubes

ten

approxi­

strain

and m i c r o s p o r e

I and

the

of

involving

21 L|_- 1 . 3 o r

21 Ip. 1 . 1 o r

cultures

grown

exception,
of

strain

experiments,

Tables

opposite

conducted

The m i c r o s p o r e - p r o d u c i n g

of p r o to p e r ith e c ia

pension

either

of

The p l a t e

serial,

prepare

were

strains

then prepared

The

trichogyne-spermatium

on c u l t u r e s

approximately

were

II.

of

rings

a Howard C o r p u s c l e

Section
was

series

106. 1 . 4 > we r e
at

of

approximately

constant

agar

cultures

of microspores

inside

was

were

plate

d a ta which might

in each

petri

in

over
sus­
of

dishes

employed.

In

the
were
the

2k

TABLE

I

EFFECT OF MICROSPORE CONCENTRATION OF STRAI N 1 0 6 . 1.1+
ON P ER I T HE C I AL PRODUCTION OF STRAI N 1 1 3 . 1 . 1
Total

Number

0
Plate

of M icrospores
80

8

800

used

to

8,000

S p e r m a t i ze +
80,000

No.

1

10

2

28

23

2

-

-

-

2

-

3

-

-

-

-

-

u

-

-

-

-

-

-

= no p e r i t h e c i a

■>:-

- no

+

= amount

culture
of

Medium = D i f c o

present

s p e rm a t i zed
liquid

used

c o r n meal

to

agar

spermat ize

(O.i+cc.)

25

TABLE I I
EFFECT OF MICROSPORE CONCENTRATION OF STRAI N 211*. 1 . 1
ON P E R I T HE C I AL DEVELOPMENT OF STRAI N 212*. 1 . 3

I
T o t a l Number

of M icrospores
120

1, 2 0 0

to

Spermatize+

0

2

-

-

2

-

1

1

66

2

-

-

-

-

-

9

12*5

3

-

-

-

-

-

16

103

k

-

-

-

-

-

2

1

5

-

-

-

-

-

1

3

6

_

_

_

—

h

16

P 1a t e

12

used

1 2, 0 0 0

1 20, 000

No.

1

+-

=

no p e r i t h e c i a

=

amount

Medium

=

of

Difco

present

liquid

used

corn meal

to
agar

spermatize

(0,2cc.)

26
one

exception,

ticular

microspore
In

a

suspension
the

In
controls

of

series

under

the

of

plates

that

perithecia.

rings

even

was

as

p e r i t h e c i a was
numbered

T a b 1e s

I and

in r e p l i c a s

were

illum ination
examined.

glass
or

for

the

of
each

eighty

to

to

incubate

hours

after

Tables

I and

rings

fewer

concentration

when

customary

which

failed

production

thousand.

however,

approximately

six

thirty

Reference

perithecial

eight

of

in each

allowed

for

containing

though m icrospore
as

I.

of m icrospore

placed

conducted,

plates

plates

Table

par­

II.

experiments
All

this

wa s

added

to
ex­

to

Go o d p r o d u c t i o n

the

quantity

thousand

and

of

of m icro ­
above,

II .

On t h e
quantity

set

up

at

0 .2 ml.

t y p e was

Likewise

high

four,

eighty microspores

evident,

spores

than

mating

Table

were

Petri

spermat ized

experiments,

constant

approximately

trem ely poor

basis

of

these

of m icrospores

exerts

a direct

tually

produced.

in each

used,

were m a in ta in e d .

received

the

was

were

rather,

of

opposite

dilution

discloses

the

dilution
series

the

both

time

produce

t wo p l a t e s

Each p l a t e

20°C(j-°C)

which
II

later

rings.

microspore

at

only

influence
Although

placed

on

depends

to

a

toperithecia

located

in

that

true

that

every

ring

will

develop

the

the

ring

hold

results,

large

it

would

inside

the

appear
glass

rings

number

of p e r i t h e c i a

even­

quantity

of p e r i t h e c i a

formed

on t h e

of pro-

measure

area,

it

does

not

number

necessarily

p r o t o p e r i t h e e i a 1 fundament w ith in

into

a mature

that

perithecium.

Rather,

the
it

is

27
believed
ogynous

that

only

those

extensions

have

more m i c r o s p o r e s

of

being

and

fertilized

dilution
tity
to

of

insure

the

sexual

the

t o make
base

of

contact

p h i 1a w h i c h

normally

growth

when p l a c e d

growth

suspension
sterile

ary

on

and

are

control

to
be

the

added

have

on c o r n meal

rings

incubators

not

the

was m a i n t a i n e d

in

with

microspores

and

incapable

at

other

of Neurospora

in­

sito-

and p r o d u c i n g
of

germination

by mycelium,

Reproduction

of

to

determine

germinating

strain

2 l i p. 1 . 1 w a s

co rn meal
Petri

of P e t r i
this

this

of

agar

plates

and c u l t u r e d

in

of

in o r d e r

m ycelium or

One m l .

exception

constant

the

agar.

sterile

a great

rings

germinating

are

capable

contact

(?)

been conducted

on s t e r i l e
the

With
was

of

the

on t h e

or

quan­

of

occupied

capable

are

one

that

m ajority

Backus

with

trich-

The

into

of

agar,

to

whose

perithecia.

coming

in Asexual

were

mature

microspores

capable

type,

indicate

dormant

already

studies

inside

30°C.
light

into

s ome

lie

sterile

treatment

triplicate

of

of microspores

glass

this

20°C,

must

shown t h a t

microspores

mycelial

and

cells

of M icrospores

whether

28

appear

In an a r e a

Cultural

ing

data

contact

mating

many p r o t o p e r i t h e e i a .
have

b . Role

developing

Apparently

vestigators

mycelial

in d i r e c t

opposite

possibility

trichogynes.

fail
the

male

c o me

the

experiment

p ro to p e rith e e ia 1 bodies,

and p r o d u c i n g
an

12,

dishes

experiment.

experiment

placed

plates.

were

at

aqueous

by

inside
Follow­

placed
16,

in

20,

cultured

Z\ \ ,
at

The c u s t o m ­
spermatizing

28
protoperitheeia
cultures

of

contained

strain

The P e t r i
and

then

lack

cultures

at

dish

under

microspores

failed

. produced

on th e

microspores

high
to

growth

whether

under

power

control

for

t wo w e e k s

revealed
the

Petri

tem peratures.

revealed

that

the

However,

of

the
dish

Microscopic

m ajority

of

s o me h a d p r o d u c e d

could

volving
5 ml .

over

to

as

received

microspores

only
be

as male

of

one

the

of

of

placed

on a h o r i z o n t a l

series

microspores

of

temperature

var.

the

autoste i r a

( G.

in

fertilization,

function

as m i c r o c o n i d i a

strains

broth

per

21 l_p. 1 . 1 ,

in o r d e r

to

from

2 11;. 1 . 3 ,

b r o t h were
located

a control

illustrate

in f u n c t i o n i n g

in­

flask.

on a s h e l f

To m a i n t a i n

up

containing

c o rn meal
or

set

containing

suspension

shaker

room.

viable

flasks

c o r n meal

containing

experiments,

used were

were

whether

e x p e r i m e n t s was

three

The

a constant

to

of water

and 2 2 7 . 3 . 7 .
either

flasks

determine

cells

Erlenmeyer

IpO m l .

of

ascospores

conditions.

series

5 nil.

to

calospora

induced

seventy-five

a s much

flask

wa s made

environmental
a

typical

plates.

G e I a s i no s p o r a

they

certain

containing

attempt

Accordingly

this

on any o f

germinate.

auto ste ir a ) fu n c tio n e d

in

on e i g h t - d a y - o I d

incubated

examination

incubation

perithecia

A further

Each

rings

germ t u b e s .
Mature

or

were

Microscopic

various

examination

short

cultures

of m ycelial

the

glass

211;.1.3.

examined.

complete

inside

as

that

in
the

spermatizing

29
cells

and

agar,

2 ml,

corn

incapable

meal

cultures

of

All

with

days

or

in

corn
the

meal

flasks

culture

strains
dishes

were

incubated

inoculated,

and were

flasks

easily

and

2 1 1 ;. 1 . 3 ,

sowed w i t h

However,

this

was

obtained

belonging

to

agar

mating

ascospores
of m ycelia,

be

it

expected
namely

appears
in

least

nA . "

calospora

stationary

var.

inoculated

w i t h ■m i c r o s p o r e s

used,

produced

also

were

c o r n meal

the

eye.

obtained

from

of

in P e t r i

2 0 ° C( j _ l ° C )

at

eventually pro­
from f l a s k s

211;. 1 . 1

t wo

in

and

this

of

hyphal
are

flasks,

from one

were

the

strands

autos te ir a

strands

mating.

strains

basis

flasks

broth

hyphal

unaided

obtained

On t h e

culture

appeared

together

these

that

contain­

microspores

strains

since

shake

type

were p r o d u c e d

certain

the

G e 1a s i n o s p o r a

of

were

strains

were

227.3.7.

at

strands

incubated

The

three

or

flasks

by t h e

mating

autoste i r a ) .
likewise

day old

rings

for

shake

hyphal

planted
and

microspores

type,

produced

to

were

typical

the

sowed w i t h

No p e r i t h e c i a

s a me m a t i n g

and m y c e l i a

glass

20°C(+1°C)

several

eight

211;.1.1

discernible

opposite

c o r n meal

was made.

results

of

after

originally

An a d d i t i o n a l

the

at

b r o t h were

227.3.7,

of

strain

hours

containing

originally

either

thirty-six

containing

duced.

of

onto

of

contained withing

on c o r n meal

longer.

211;.1.1

perithecia

growth

s u s p e n s i o n was p o u r e d

2 1 1 ;. 1 . 3

Wh e n t w o m y c e l i a
shake

mycelial

Also p r o to p e r ith e e ia

0 .2 ml.

plates

W ithin
ing

plates.

strain

spermatized

eight

microspore

agar
of

of producing

of

those
( G.
which
the

and m y c e l i a l

30
growth
of

inside

the

approximately

would

appear

flasks

s o me

eral

environmental

play

an

the

important

plates

of

typical

fact

broth

resulted

celia

belonging

this

of

reproducing

sp.

in

and o t h e r

or

than

plates

production

Gelas inospora

presents
do

the

strong

function
organism

ascomycetes,

Therefore,

aeration
the

or

of

it

sev­

o x y g e n ma y

germination
received

sterile.

period

of

2 ml,

the
of

The c o n t r o l

spermat ized produced

rings.

microspores

the

incubation

a combination

which

remained

the

an

longer.

influencing

within

to

organism

factor

other

control

that

a u to s te i r a ),

or

211p.l.3 which were

perithecia

of

in

suspension

strain

The

( G.

role

after

hours

other

factors

Those

microspore

only

forty-eight

that

microspores.

but

when p l a n t e d
of

hyphal

calospora var.
evidence

to

those

providing

c o r n meal

strands

that

as m i c r o c o n id i a
as

in

autoste ira
the

and

produced

cultural

a n d my ­

microspores

are

capable

by Neuro spor a

conditions

are

su i t a b l e .
Since

the

evidence

tial,

attem p ts were

sion.

Germinating

attached
on

were

sterile

develop

retrieved

tions

side

further

from b r o th

agar.

germ t u b e s

development,

to

m icrospores with

corn meal

long

producing

ma d e

submitted

Qp m i c r o s p o r e s

admittedly

substantiate
the

for

shaker

a short
of

the

In over

seeded

in

time

circumstan­
this

spore w all
flasks

The m i c o r s p o r e s

H o w ever, none
branches.

is

still

and p l a c e d

continued

and

then

germ t u b e s

conclu­

were

to

ceased
observed

t wo

thousand

slide

prepara­

liguid

culture,

a germinating

31
microspore

possessing

observed.

M icrospores

produce
spores
was

m ycelial
actually

relatively

sands which
of

spores

which

were

small

stages

placed

failed

to

up

filter
but
to

of

into

dormant.

to

paper.

failed

to

reiterate

develop
that

the

of

microspores

on

or

in

potential
with

possibility

a single

one would
rather

side

t wo

actually

medium.
of

to

thousand

to

of

These

agar
flasks

germ t u b e s
importance
of

the

germ t u b e s
this

the

microspore

small.

thousand

total

either

fact,

a germinating

extremely

also

moistened

extreme

proportion

a hundred

catch

solid

produced

In view of

prepare

on

m ic ro s p o r e s were

produce

discovering

a

Som e m i c r o ­

containing

small

thou­

growth.

the

is

of

Perhaps

slides

this

particular

stage

that

any

if

of

germination.

Dodge

(11)

has

pointed

of producing

a germ tu b e

of

vegetative

producing

d it ions.

It

b r a n c h w o u ld be

even have

than

microspore

liquid

of

germ t u b e s

illustrates

flasks.

microspores

further.

quantity

germ t u b e s

dishes

an e x t r e m e l y

2

to

of m icro ­

hundreds

figure

been

failed

number

the

of m ycelial

Petri

Som e o f

to

culture

drop p r e p a r a t i o n s
inside

the

never

agar

microspores.

produce

shake

has

and p r o d u c i n g

Text

germinating

show a n y e v i d e n c e

and k e p t

solid

Furthermore

in com parison

liquid

branch

on m o i s t e n e d

germinating

commenced

Hanging
set

found

side

planted

growth.

remained

series

a single

out

should
mycelium

also

spore,

be p o t e n t i a l l y

under

certain

capable
capable

cultural

con-

32

Text

Figure

2.

V arious Stages of Successive Microspore
G e r m i n a t i o n on S o l i d A g ar as drawn by
Camera L ucida.
c a , x 1000.

33
It

appears

strange

s P Qra c a l o s p o r a v a r ,
are

capable

of

to

ical

environmental

on

produce

the

autosteira

producing

able

that

long

m ycelial

( G.

is

germ t u b e s ,

on a g a r .

not

of G elasin o -

a u t o s t e i r a ) , s o me

unbranched

growth

factor

microspores

Perhaps

supplied

to

the

of which
are

s ome

un­

c rit­

microspores

agar.
Wh e n

the

microspores

shake

and

stationary

celia

representative

flask.

Therefore,

circum stantial,
able,

at

ing

as

the

agar

least

culture,
of

it

that
under

problem

not

should

placed

in c o r n meal

a few g e r m in a te

appropriate

appears

certain,

microspores

certain

an

investigated

broth.

in
my ­

in each

admittedly

fungus

conditions,

inhibitor

c o r n meal

type

although
this

broth

and p r o d u c e

mating

of

cultural

Perhaps

in the

be

the

the

m icroconidia.
and

are

are

of

cap­

function­

may b e p r e s e n t
This

in

interesting

further.

B . M igrating Nuclei
Sexual
autoste ira
actions,
of

nuclei

reproduction

( G.

autoste i r a )

namely
of

the

opposite

opposite

m icroconidial

type

is

an

least

the
step

interaction

resulting

relationship

w hereby plasmogamy

in

prelim inary

upon

at

has

initiated

calospor a var.
t wo

of p r o t o p e r i t h e e i a

m a t i n g —t y p e

the

depends

mating

involves

production

Furthermore,
reproduction

i n G e 1a s i n o s p o r a

in

re­

fusion

ai>c i .

leading

sexual

to

between m ycelia

suggested
this

and

of

developing

in plasmogamy.
been

types

of

A trichogyne
as

fungus.

the

mechanism

3lj_
With

this

particular

w as made

to

okaryons

involving

produced

in

in

this

this

fungus

conidial
not

determine

of both
The

whether
mating

fungus.

to

sexual

and

not

of

somatic

mating-types

reproduction

agency of
by

an a t t e m p t

could

is

to

be

occur

a trichogyne-micro-

somatogamy,

dicaryotic

heter-

then

mycelia

it

should

comprising

mating-types.

compatible
to

If

the

in m in d ,

production

opposite

isolate

experiments

type

the

of

through

relationship,

be p o s s i b l e

nuclei

whether

nuclei

only

hypothesis

to

be

nuclei

those

of

described

were

could m igrate

another

giving

ma d e

to

determine

from hyphae
rise

to

of

one

dicaryotic

my c e 1 i a .
A number
which

of

two p e r c e n t

the medium c r o s s
each

experiments

co rn meal
arm and

a g a r was

of

u-tubes

each

were

u-tube

treated

were

in

of both

strain

ing

strain

Thirty-five

prepared
inside

211; . 1 . 1 .

and p l a c e d

a 20°C(±1°C}

under

A few days
of

observed

the
to

inside

as

the

the

g r o w d o wn t h e

or

such

a moist

for

after

u-tubes,

211; . 1 . 3

incubator.

observation

such

out

in

poured

to

fill

arms

of

upright

a manner

inoculated with

protoper i thee ial

arms

carefully

one-quarter

the

kept

was c a r r i e d

u-tube.
All

arms

u-tube

long

chamber

as

arms

from the
and

cultures

u-tubes

were

were

months.

w e r e made
opposite

appeared

produc­

w h ic h was p l a c e d

these

three

both
either

microspore

u-tube

inoculations

hyphae

mycelium of

the

Some o f

that

in th e
arms were

to meet

somewhere

35
in

the

middle

by

aid

of

the

in

the

but

arms

of

at

a

small

microspore

Perhaps

no

Plate

also,

took p la c e .
would

raise

time

one

space

air

a few

Two m o n t h s

later

the

medium arm r e g i o n

and

a small

chunk

of

center

of

plates.

These

plates

were

ten

days,

mature

produced

that

interm ingling

had

occurred

in

in

over

the

A A.

hyphae

appeared
offer

of

t o ma k e

from

contact.

an e x p l a n a t i o n

as

to

did develop

int h i s

u-tube

allowing

subsequent

fail

to

of

in term s

with

of

these

the

m e d 1 um a r m o f

were
aid of

from t h i s

at

somatogany

broken

region

agar

in

a small

20°C(+1°C).

and

W ithin

typical

ascospores

plates

indicating

of

the

file

in P e t r i

t wo c o m p a t i b l e

s ome

in

further?

u-tube

c o rn meal

of

copulations

protoperitheeia

u-tubes

containing

of

the

develop

incubated

discernible.

hyphal

the

sterile

one-half

sperm atization.

conspicuously

to why d i d

hyphae

coming

pro­

u-tube.

and

perithecia

were

the

was t a k e n

on t h e

presumably of

solitary

each

planted

revealed

in one

s o me

agar

u-tubes

did develop

as

of

the

no p e r i t h e c i a w e r e

an e x p l a n a t i o n

u-tubes

of

usually

hyphae

trichogyne-vegetative

question
the

to

s p a c e was

To o f f e r

arm o f

had

of

figure

exception,

development

the

111,

u-tubes

this

air

such

microscope

u -tu b e s where

the

Observation

intermingling

r e a s o n why p e r i t h e c i a

However,

the

s o me

the

u-tube.

binocular

a single

difficult

Perhaps
for

each

types,

any of

opposite
is

of

mating

With
duced

of

a dissecting

occurrence

opposite

It

band

strains

u —t u b e s .

36

PLATE 111

F i g u r e s A- B

u-tube

F i gure

This

.

and

hyphal

represents

inoculated
211+. 1 . 3
were

F i g ur e A ^ .

F i gure

B.

of

of

the

one

exception,

in

these

strain

211+. 1 . 1 .

Perithecia

containing

outermost
tip

perithecia.
ithecia

plates

latter

plate,

strain

no p e r i t h e c i a

one

arm o f w h i c h was
and

suspension
developed

microspore

represent

the

other

strain
in

those

suspension.

four

derived

which

of

only

failed

Numerous p e r i t h e c i a
plate

strain

arm w i t h

211+.1.3

subcultures,

center

in the

the

u-tubes

u-tubes.

u-tubes,

a microspore

inner

arms w i t h

With

several

hyphal

thirty-five

opposite

arm w i t h

The

the

the

inoculated with

arms

experiments.

in

produced

One o f

one

in one

and

211+.1.1.

tip

of

the

from the
to

develop

developed

but

only

after

protoper-

and m i c r o s p o r e s

made

their

appearance.

37

38
Since
in

the

the

bend

tubes

dry

were

of

the

at

in

forming

perithecia
should

perithecia

were

originally

develop

the

been

surface

was

2 U4. . I . 3 ,

conidia

eventually
in

old

air

drying
the

to

up,

no

u-tubes.

be

capable

of

time.
at

in

this

time,

those

inoculated

arms

with

however,
of

the

the

that

no

u-tubes which

protoperitheeia 1

in

arms,

failed

these

to

pockets

produced

of

series

and

strain

u-tubes

g r o w n down
its

the

the

placed

t wo

inside

days,

structures

around

appearance

a mycelial

mat

at
of

arm w i t h

The

on

to

a suspension

a 20°C(+°C)

kept

incubator

originating

the

one

arm o f

a protoperitheeial

u - tu b e s were

the middle
the

experiments,

w ith mycelium of

hyphae

inoculated with
and

u-tube

other

211).. 1 . 1 .

illum ination.
W ithin

of

inoculated

stant

ing

agar

too

the

allowed

pro to p er i thee I a were

chamber

made

though

agar

in

a v a i l a b 1e f o r

Even

on t h e

forms

Although

moist

the

this

to be

pocket

form

further.

u-tube

strain

at

perithecia

u -tu b es were

m y c e l i u m was

produced

In a n o th e r
each

produced

be n o t e d

211j.. 1 . 3 .

agar

the

the

that
an a i r

oxygen

s o me o f

tubes,

ever

however,

the

o n l y when

sufficient

the

Perhaps,

strain

suggested

room t e m p e r a t u r e .

formed

It

has

u-tubes

formation,

p e r i t h e c i a were

had

(10)

allowing

per i thee ial
air

Dodge

of micro-

inside
under

from the

surface

hyphae w i t h i n

of

the

the

the

u-tubes

opposite

microspore

con­

arm o f

protoperitheeia 1 strain
arm o f

a

had

and had
arms

form­

suspension.

39
Shortly
ance

on

those

ithecia

hyphal

developed

ascospores.
arms

afterwards

of

The

the

into

u-tubes

their

E v e n t u a l l y many o f

mature

perithecia

originally

strain,

ma d e

the

containing

produced

in

the

inoculated with

failed

to

develop

appear­

protopertypical

opposite

hyphae

further,

of

the

Plate

III,

.
It

growth

mats.

protoperitheeia

protoperithecial
figure

pro t o p e r ith e e ia

is

of

special

issuing from

interest

the

to

note

microconidial

that

no m y c e l i a l

suspension

could

be

detected.
On t h e
that

fusions

opposite

basis
or

mating

of

the

above

anastomoses
types

results

occurring

do n o t

mating

types

in

this

Dodge

(12)

Neurospor a nuclei
apart

until

opposite

of

states
of

that

opposite

and C o o n r a d t
somatic

hyphae

containing

bodies

(perithecia)

the

the

of

fundamental

receive

nuclei

of

mech­
opposite

fungus.
in h e t e r o t h a l 1ic

species

mating

normally kept

occurence

of

probably

nuclei

hyphae."
the

(8)

types

suggest

heterokaryons

mating-types

vegetative

certain

are

of

reproduction.

Beadle
formation

appears

between m ycelia

represent

anism w h ereb y young p r o t o p e r i t h e e i a

it

of

rather

is

the
than

T h i s may be
one

that

between
due

t wo

to

in

t wo

the

sexes

grow

to

failure

in the

of

strains

of

the

"tendency

of

the

form f r u i t i n g

the manner

a possible

exception

the

of

strictly

explanation

u —t u b e

for

experime n t s *

40
However,

an e x p e r i m e n t

now t o

be

described

tends

t o ma k e

this

an 1 i k e l y .
In
nuclei
type

a further

could

to

be

those

compatible
ithecial

to

compatible

of

hyphae

ular

the

plates

hours

after

transferred

both mating
at

experiment

strains

hyphae

the

had

been

discs
types

dishes

time

sterile

the

hyphal

on t h e

of

hyphal

were

Petri
of

and

produced
c o r n meal

anastomosing

intermingling

tips,

Intermingled
to

c o r n meal

agar.

twenty-four

were
Of

binoc­

transferred

for

tips

isolated

the

and

ninety-nine

none p r o d u c e d

containing

hyphae

numerous m ature

plate,
this

inoculated

and

sterile

culture

isolated

t wo m a t i n g

containing
out

were

a dissecting

plates.

produced

experiment

the

Wh e n

2 0 ° C( + _ 1 ° C)

mixed

young p e r i t h e c i a

inoculated

at

corn meal

however,
of

cut

containing

from th e s e

periphery

agar

were

single

original

types,

of

aid

whether

together.

intermingling

the

one m a t i n g

of p ro to p e r-

tips

type

hastened.

with

incubated

In a second
containing

be

this

of

region

and 2 1 i | . . l . l ,

that

would

small

which

The

so

evident,

were

of

211|.1.3

another

developed

perithecia.

the

types

first

to

in

compatible

investigate

both mating

of P e t r i

These

cia

localized

both mating

center

cultures

become

to

containing

became

of

as

hyphae

hyphal

microscope,

hyphae

somatic

t wo h u n d r e d

one

of

whether

over

namely,

adjacent

from

as w ell

nuclei

the

determine

another,

formation,

In

to

transferred

of

from c u l t u r e s

types,

attempt

Plate

type,

III,

perithe­
figure

B.

agar

block

from mixed m y c e lia ,

which

agar

a small

of

ten

days p r e v i o u s l y ,

1+1
was

placed

agar.

in

the

The P e t r i

constant

illum ination

the

were

co rn meal

agar

of

these

at

20°C(+^1°C)

aqueous
mature
in

of

between

opposite

ible
of

the

occur

in

sinospora

tips

types
to

results

concept

agency

to

the

the

tubes

hyphal
base

after
for

to

hyphal
that
the

of

on s t e r i l e
none

incubation

several

weeks.

0.5 ml.

strain

a s c o s p o re s were

discover

211+.1.1,
produced

tips

of

both mating

mycelia

of

both mating

perithecium
for

and c o n s e ­

hyphal

anastomoses

a dikaryon produced

tends
occur

Mor e

tend

six

occur.

in the

that

var.
of

of

isolated

only

even

available

necessarily

calospora

the

base

development.

The

aforementioned

the

however,

associations

trichogynes.

through

at

do n o t

perithecial

illustrating

mating

hyphal

nuclei

that

o p p o r t u n i t y was

Failure,
any of

indicate

under

experiment,

belonging

tubes.

test

and

spermatized with

these

present

test

illum ination

were

corn meal

20°C(+1°C)

from

previous

typical

types were

out

in

containing

retrieved

ample

the

at

hundred

growing

of m icrospores

types were

containing

hours.

perithecia

constant

results

quently

in

cultures

perithecia

These

As

produced

suspension

one

each was p l a c e d

under

forty-seven

twenty-four

procedure,

slants.

Wh e n t h e s e

dish

incubated

from hyphae

and

cultures

a Petri

then

for

this

obtained

per ithecium

of

d i s h was

Following
tips

center

to

to

suggest

localized

likely,

that

compat­

around

compatible

areas
nuclei

a s c o g o n i a by way o f
further

plasmogamy

autoste ira
functional

( G.

is

the

substantiate
initiated

the

in G ela-

a u t o s t e i r a ) only

sexual

from

structures.

SECTION IV
DI S CUS S I ON AND CONCLUSION
Function
Numerous
Olive

(30)

and

of

fungi

have

others,

to

the

been
be

under

certain

in most

of

cases

reported,

well

tests

as

microspores
In
function
spora

to

have

this
of

determine

in the

ship

Microspores
function

immediate
2.

a.

conditions.

specific

function

these

of

a n a t t e m p t w a s ma d e
bodies
as

to

determine

produced

as

reported

by T y l u t k i

(free

of hyphal

sexual

fragments)

cells

have

(spermatia)

of

a trichogyne-microspore

in t h i s

is

no

numerous

this

organism

still

assumption

evidence
u-tube

of

are

relation­

questionable.

the

following

facts:

somatogamy

indicated

by

experiments

k2

remains

been

when p l a c e d

The p o s s i b i l i t y

of

(35) .

Sexual R eproduction

as male

in fa v o r

the

by G e l a s i n o -

protoperitheeia.

the

form

tests

of

There

by

However,

vicinity

functioning

However,

s ome

cytological

s p e r m a t i u m - 1ike

I.

shown t o

literature

adequate

calospor a v a r . autoste i r a ,

1.

the

of producing

cultural

the

in

been n eglected.

study,

the

reported

capable

of m icrospores
the

Microspores

reported.

b.

There
in

is

this

no

evidence

organism

subcultures.
taken

at

further
c.

The
is

d.

the

correlated
the

the

of

distribution

have

to

a direct

tive
In

that

different

capable

expect

to

However,

particular

tion

in

in

of

developing
areas

used

to

upon

the

could
is

of

perithecia

the

cultures

as w e l l

as w i t h

spermatize
number

indicate

appears

of p e r i t h e ­

that

n e e d e d when

s ome c o me

approach

a

large

spermatizing

in c o n t a c t w i t h

if

the

evidence

we d i d

point.

problem,
in

the

throughout

somatogamy were

find

homothallic
the

to

of m igrating

the

recep­

nuclei

s a me m y c e l i u m ,
the

mycelium.

functional

of n u c l e a r

of

we

should

migrations.

not.

and A l e x o p o u l o s

this

the

point.

t y p e s when p r e s e n t

Therefore,

Wilson

tips

tr ichogynes.

a negative

are

tip

protoperitheeia.

of m icrospores

insure

in hyphal

microspores

effect

This

by hyphal

developing p e r ith e c ia

with

of m icro sp o res

produced.

number

this

pattern

spermatized with

Quantity

dicaryons

of young

substantiate

distribution

cia

detected

base

d ikaryo t i zat ion e x i s t i n g

demonstrated

Lack of

directly

to

e.

as

of

(36)

Mutant
fungus,

heterokaryotic

have

nuclei

recently

affecting

illustrated
spore m atu ra­

G elas inospor a c a l o s p o r a , were
mycelium,

hyphal

tip

transfers

yielding
gating

colonies

perithecia
Thus,

of

which produced

apparently

Either

and

represents

there

this

certain

that

hyphae

Because
in

attempting

attachments
tance

of

ganism,
of

order

thecial

reported

fungus

occurs

the

the

determine

investigate

result

s o me
of

formation

stim ulation.
perim ents

hormone

(5)

the

action

(10)

paper,

in

the

dissected

and

appears

nuclei

ascogonia

relations

in the
be

of

in

in
and

life

rather

or­

a refinement
in

thoroughly.

sexual
state

cause

bodies

fusion

that

repeated

of hormonal

the

fruiting

is

study

impor­

level,

more

of

(26)

(5)

of

toward

than

s i t o p h i 1a

vital

cycle

aspect

formation

this

of microspore

the

directed

and A r o n e s c u

no e v i d e n c e

place.

it

encountered

and Moruzi

in N eurospora

Dodge

takes

trichogynes

trichogynes.

particular

Moreau

and found

Aronescu

this

relationship

fertilization

the m icrom anipulator

authors,

nuclei.

only

and b e c a u s e

should
at

this

possible

trichogynes,

possibly

autosteira

of compatible

difficulties

research

to

segre­

of mechanisms

between

strains

in

association

future

compatible

fact

the

copulation

compatible

point

to

the

of

particular

According
is

frequent

this

techniques

theories

a microspore-trichogyne

by way o f

of
to

to

two v a l i d

o n ly method by which

results

h e t e r o t h a l 1ic

ascogenous

or

hyphae

From t h e
quite

is

the

accomplished

and v e g e t a t i v e

only

i n G e 1as i n o s p o r a c a l o s p o r a v a r .

remain.

can be

and

randomly d i s t r i b u t e d .

fertilization

this

non-segregating

of

peri-

by hormonal
these

ex­

stim ulation.

In

grew a s c o s p o r e s

from

single

1+5
asci

and

types

in

in

a

every case

found

1:1 m e n d e l i a n

In. G e l a s i n o s p o r a
(3l+)

in

a genetical

demonstrated
fusion

of

that

as

were

analyzed.

found

then

all

type.

in

Both,

this

two n u c l e i
action,
of

the

type.

secutive

days

should

Control

likewise

result

taken

hundred

type

of

nuclear

randomly

as

and f o r t y - t w o

and p i g m e n t a t i o n
fashion.

stimulated

theoretically

demonstrate

mating

be

that

the

types

e x p e r i m e n t s was

If

of

asci

loci

were

sexual

by hormone

re­

action,

one m a t i n g

perithecial

result

of

suspension

fusion

of

due

set

in which p o r t i o n s

up,

of microspores

of

to

for­

and n o t

a p r o t o p e r i t h e e i a 1 s t r a i n were

treated

the

p ro d u c e d by t h e s e

the m icrospore

hormonal

with

opposite
cultures.

suspension prior

developed p e rith e c ia .

experiment,

plates

when 0 . 2 ml.

exposed

ten minute

contained w ithin

develop.

t wo m a t i n g

so.

previously
for

a direct

mendelian

treated with

another

is

s p e r m a t i z a t i o n was

filtration

microspores,

1:1

further

of

the

organism has

fro m one

No p e r i t h e c i a w e r e

plates

In

thecia

of

filtered

Seitz

a

of

autoste i r a , Tylutki

Ascospores

mating

opposite

colony

Control
to

of

a series

a Seitz
mating

to

following

this

organism were

was n o t

In o r d e r
mation

in

ascospores

This

of

nuclei.

taken

segregate

production

calospora v a r .

fertilization

ascospores

to

segregation

ratio.

study

compatible

well

a

glass

to

u.v.

periods,
rings,

suspensions

light

was

for

added

perithecia

developed mature

to

three

of
con­

protoper i-

failed

perithecia.

to

ke
Four
were

allowed

cular

disks

colonies
on

the

the

cultures
to

produced

young

These

by th e

be

culture

growth

is

these

if

returned
of

to

the

type,

type

and

i t would be

into

results,

it

into

to

hormone were
stimulated

expected

in one

appears

tend

arm o f

the

that
the

development.
unlikely

becoming mature

that
perithe­

action.
Asexual

been

Reproduction

definitely

shown t h a t

c o n t a m i n a t i o n s when p l a c e d

the

production

flasks

the

attempts

these

also

a diffusible

one m a t i n g

stimulated

within

furthermore

is

For,

earlier

perithecial

has

to

mentioned

stim ulated

are

in

liquid

micro spore

cir­

present.

produced

of

it

then

protoperitheeia

from m y c e lia l

All

examination

mating

II.

corresponding

later,

results.

by any hormonal

result

days

experiments

other

protoperitheeia

Again

were

mycelium of

the

over

plates

no p e r i t h e c i a

above

placed

Petri

Forty-five

developing

ture,

sterile

o f m i c r o s p o r e s was p l a c e d

In view

free

Then

suspension

■u-tubes would

cia

seven days.

an aq u eo u s

11- t u b e

mycelium of

pro toper i thee i a 1 s tr a in s

were

revealed

the

for

the

cellophane

incubator.

support

of

of w ettab le

and

The

one

incubate

cellophane.

plates

of

of m ycelial

forty-eight

demonstrable

of

the

microspores
in

liquid

cul­

growth w i t h i n

hours.
mating

the

This m ycelial
type

strain

inoculum.
to

capable

demonstrate
of

going

conclusively

on and p r o d u c i n g

that

any given

mycelium,

have

k7
failed.

However,

proportion
chance

of

of

the

picking

mycelium would
On t h e
that

the

ste ira
certain
are

since

it

appears

microspores

that

actually

only
do

germinate,

a m icrospore which w i l l

be v e r y
basis

microspores

the

small.

of work

reported

of Gelas inospora
as male

here,

it

is

calospora

prim arily

as y e t

unspecified

environmental

functioning

as m i c r o c o n i d i a .

of

small

go o n a n d p r o d u c e

function

capable

a very

sexual

cells,

concluded
var.
but

conditions

auto under
they

SECTION V
SUMMARY
1. A s t u d y
the

of

the

capable
cells

of

that

that

functioning

the

cells

these

as

particular

spermatizing

is

also

structures

sexual

p re se n te d which

microspores

may f u n c t i o n

as

(microconidia)

endowed w i t h

the

var.
are

propagative

strongly

asexual

indicates

propagative

capacity

to

reproduce

organism.
2.

cross

In

connection with

sperm atization,

when m i c r o s p o r e s w e r e
development

within

ithecia

of

forty-eight

indicate

that
only

and

to

structures

operative

details

trichogynes

be w o r k e d

and

of

protoperitheeia,

perithecia

develop very

fertilized

cell

by c o m p a t i b l e

out.
1+8

does

occurred

appear

rapidly

relationship

into

appear

fusions

microspores

to
per­

microspores.

a u to ste ir a .

and n u c l e a r

compatible

development,

This would

i n G e 1a s i n o s p o r a c a l o s p o r a v a r .

cytological

between

into

seventy hours.

being

on s p e r m a t i z a t i o n ,

perithecial

on c o m p a t i b l e

protoperitheeia
after

experiments

localized

placed

these

the

A trichogyne-microspore

the

produced by

( sp e rm a t i a ).
Evidence

the

of microspores

h e t e r o t h a 11ic A s c o m y c e t e , G e la s in o sp o r a c a l o s p o r a

a u to s te ir a , revealed

the

function

still

to

be

However,

occurring
need

to

1+9
3.
and
tend

thirty-four
to

brought

that

into

close

only

over

results

compatible

associations

nuclei

probably only
Plasmogamy

through

microspore

cultures,

and n u c l e a r

Compatible

gynes.

involving

nuclei

G e 1a s i n o s p o r a c a l o s p o r a v a r .

anastomoses

fungus

study

u-tube

indicate

ascomycete

tive

In a

the

relationship

ascogonia
is

structures.
must

in

obtained

the

that

h e t e r o t h a l 1ic

a u to s te ir a are

one

in c l o s e

and karyogam y

sexual

were

hyphal

another

not

by v e g e t a ­

m igration.

occur

in the

with

t wo h u n d r e d

be

associations

b y way o f

the

in t h i s
tricho­

accomplished probably
Therefore,

operative

a trichogyne-

in t h i s

fungus.

tips

LITERATURE CITE D
1.

A l e x o p o u l o s , C.
Ge l a s i n o s p o r a .

J . a n d S . H. S u n .
1950.
M y c o l o g i a ip2: 7 2 3 - 7 3 l p .

A new s p e c i e s

2.

A m e s , L . M.
1930.
A s t u d y o f s o me h o m o t h a l l i c
o t h a l l i c Ascomycetes.
M ycologia 22: 318-322.

3.

A m e s , L . M.
1932.
An h e r m a p h r o d i t i c s e l f - s t e r i l e b u t
c r o s s - f e r t i l e c o n d i t i o n in P le u ra g e a n s e r i n a .
Bull.
T o r r e y B o t . C l u b 5 9 : 3l±l-3L&~.

of

and h e t e r -

Ip. A m e s , L . M.
193l p.
Hermaphroditism involving s e l f - s t e r i l ­
i t y and c r o s s f e r t i l i t y in the Ascomycete P l e u r a g e a n s e r ina.
M y c o l o g i a 2 6 : 392-lpllp.
5* A r o n e s c u , A.
1933.
Further studies
p h i 1a .
M y c o l o g i a 25* Jp3-5U--

in N e u r o s p o r a

sito-

6.

B a c k u s , M. P .
19314..
I n i t i a t i o n o f t h e a s c o c a r p and a s s o ­
c i a t e d phenomena in Coccomyces h i e m a I t s .
C o n t r i . Boyce
T h o m p s o n I n s t i t u t e 61 3 3 9 - 3 8 0 .

7-

B a c k u s , M. P .
1939.
The m e c h a n i c s o f c o n i d i a l f e r t i l i z a ­
t i o n i n N e u r o s p o r a s i t o p h i 1a .
B u ll. Torrey Bot. Club.
6 6 : 63~7"S7

8.

B e e d l e , G. W. a n d V . L.
i n Ne u r o s p o r a c r a s s a .

9.

C a i n , R. F.
1950.
1.
Gelas in o sp o ra .

10.

Dodge,
mones

11.

D o d g e , B. O.
m icroconidia
3 I4.7 - 3 6 0 .

Coonradt.
191+14-.
He t e r o k a r y o s i s
G e n e t i c s 29: 29 1 -3 0 8 .

Studies of coprophilous Ascomycetes.
C an a d ia n J. R e s.
29: 5 6 6 - 5 7 6 .

B. O.
1931.
H e t e r o t h a l 1ism and h y p o t h e t i c a l h o r ­
in N e u r o s p o r a .
B u l l . T o r r e y B o t. Club 58: 5 1 7 - 5 2 2 .
1932.
The n o n - s e x u a l
of Neurospora.
Bull.

and s e x u a l f u n c t i o n o f
T o r r e y B o t. Club. 59:

12.

D o d g e , B. O.
1935*
Th e m e c h a n i c s o f s e x u a l
in N e u ro sp o ra .
M y c o l o g i a 27: Ipl8-lp38.

13.

Dodge
B. O.
1936.
Pleurage anser in a .

S p e r m a t i a and
M y c o lo g ia 28:

50

reproduction

nuclear m igrations
2 8 L |.-2 9 1 .

in

51
ll+.

D o d g e , B. 0 .
B-ull. T o r r e y

191-1-2.
H e t e r o c a r y o t ic v i g o r
Bot. Club. 6 9 : 75-91.

in N eurospora.

15.

D o d g e , B . O.
191+6.
S e 1f - s t e r i 1 i t y i n " b i s e x u a l ” h e t e r o caryons of Neurospor a .
B u l l . T o r r e y B o t . C l u b . 7 3 : L|_10 —
ll-l 6 .

16.

D o w d i n g , E . 'S.
1933.
Ge 1 a s i n o s p o r a - a n e w g e n u s o f
Pyrenomycetes w ith p i t t e d spores.
Canad. J o u r. Res.
9 : 2 9 1 + -3 0 5 .

17.

D o w d i n g , E . S . a n d A . H.
t i o n i n Ge l a s i n o s p o r a .

18.

D o w d i n g , E . S. a n d A. B a k e r s p i g e i .
1958.
Poor f r u i t e r s
a n d b a r r a g e m u t a n t s i n G e 1a s i n o s p o r a .
Can. J o u r . B o t.
31+: 2 3 1 .

19*

Drayton,
conidia

20.

D r a y t o n , F . L.
1931+. T h e s e x u a l
g 1ad i o 1 i .
M y c o l o g i a 2 6 : 1 + 6 -7 2 .

21.

H i g g i n s , Bascombe B r i t t .
1920. M o rp h o l o g y and l i f e
h i s t o r y o f so m e A s c o m y c e t e s w i t h s p e c i a l r e f e r e n c e t o
the f u n c tio n of spermat ia.
Am. J o u r . B o t .
7 : 1+3 5 —1+1+lp -

22.

H i g g i n s , Bascombe B r i t t .
1929. M o r p h o l o g y and l i f e
h i s t o r y o f so m e A s c o m y c e t e s w i t h s p e c i a l r e f e r e n c e t o
the f u n c tio n of sp erm atia.
Am. J o u r . B o t .
16: 2 8 7 - 2 9 6 .

Buller.
19l|-0. . N u c l e a r
M y c o l o g i a 3 2 : 1+71-1+88.

migra­

F. L.
1 9 3 2 . The s e x u a l f u n c t i o n o f t h e m i c r o in c e r t a i n Discomycetes .
M y c o l o g i a 2l+: 3l_+5—31+8.

1893.

Monilia

fru ct igena.

Sc l e r o t i n i a

23.

Humphrey, J.
18: 8 5 - 9 3 .

2l+.

L i n d e g r e n , C.
1 9 3l+.
Genetics of Neurospora w ith s e lfs te r ile bisexual heterokaryons.
J o u r . G e n e t i c s 28:
1+25-1+35.

25.

L i n d e g r e n , C.
1936a. H e t e r o c a r y o s i s and hor mone s
Neurospora.
Am. N a t .
1 2 0 : 1+01+-1+05.

26.

M o r e a u , F . e t C. M o r u z J .
1936.
Recherches sur la
g e n ^ t i q u e d e s A s c o m y c e t e s du g e n r e N e u r o s p o r a .
Rev.
g £ n . de B o t .
1+8: 6 5 8 - 6 7 5 .

27.

E.

mechanism of

M o r e a u , F.
1939.
Sur l e s m i c r o c o n i d ie s des
B u l l . S a c . B o t . F ra n c e 86: 12-16.

Bot.

Gaz.

in

Neurospor a .

52
28.

Moreau, Claude e t M i r e i l l e .
19ij-9.
du Congo.
Supplement C olonial a
Tome X I V , N o . 2 : 5 0 - 6 6 .

Quelques Ascomycetes
l a R e v u e de M y c o l o g i e .

29.

Moreau, Claude e t M ireille^.
1951Le g e n r e Ge 1 a s i n o ­
spor a Dowding.
La M y c o t h e q u e , P r e m i e r S u p p l e m e n t .
M i c r o m y c e t e s 39 -1 + 1 .

30.

O l i v e , L. S.
1953.
The s t r u c t u r e and b e h a v i o r
fungus n u c le i.
B o t . R e v i e w 19: 1 + 39-586.

of

31.

S a n s o m e , E. R.
191+6.
Heter okaryos i s , m ating-type f a c ­
t o r s and s e x u a l r e p r o d u c t i o n in N e u r o s p o r a .
Bull.
T o r r e y B o t . C l u b . 7 3 : 397-1+09.

32.

S u n , H. S . 1 9 5 6 .
A c o m p a r a t i v e morpho lo g i c a l and c y t o l o g i c a l s t u d y o f G e l a s i n o s p o r a c a l o s p o r a (Mouton)
M o r e a u e t M o r e a u , G e 1a s i n o s p o r a a u t o s t e i r a A l e x o p o u l o s
e t Su n an d G e I a s i no s p o r a c e r e a l i s D o w d i n g .
P h . D.
T h e sis, Michigan S ta te C ollege.
56 pp.

33.

S u n , H. S . , C o n s t . J . A l e x o p o u l o s , a n d G. B. W i l s o n .
1951+.
A c y t o t a x o n o m i c s t u d y o f t h r e e s p e c i e s o f Ge 1 a s i n o s p o r a . C y t o l o g i a 19-* 2 5 5 -2 6 1 ]..

3l+.

T h a x t e r , R. 1 9 2 6 .
C o n t r i b u t i o n s towards a monograph
the Laboulbeniaceae I-IV.
Mem. A m e r . A c a d . A r t s a n d
S c i . 15 - 1 + 3 1 - 5 8 0 .

35.

T y l u t k i , E. E.
1955*
A s t u d y o f som e a s p e c t s o f m o r ­
p h o l o g y g e n e t i c s and c u l t u r a l b e h a v i o r o f t h e h e t e r o t h a l l i c pyrenomycete Gelas inospor a c a lo sp o ra var.
a u t o s t e i r a ( A l e x o p o u l o " i e~E S u n ) A l e x o p o u l o s a n d S u n .
P h . D. T h e s i s .
Michigan S ta te U n iv e rsity .
73 p p .

36. W h i t e h o u s e ,
the fu n g i.

H. L . K.
191+9.
He t e r o t h a 11 i sm a n d s e x i n
B i o l . R e v . C a m b r i d g e P h i l . S o c . 21+: 1+11-1+1+7.

3 7 . W i l s o n , G. B. a n d C. J . A l e x o p o u l o s .
m u t a t i o n i n G e 1a s i n o s p o r a c a l o s p o r a ,
f ungus .
M yco 1 o g i a
1+8: 6 6 5 - 6 8 9 «
38.

of

1956.
Spontaneous
a homothallic

Z i c k l e r , H.
1931+.
G e n e t i s c h e U n t e r s u c h u n g e n an e i n e n
H e t e r o t h a l 1 i s c h e n a s k o m y z e t e n ( B o m b a r d 1a l u n a t a N o v .
spec.).
P l a n t a 22: 5 7 3 - 6 1 3 .

39.

Z i c k l e r , H. 1 9 3 7 .
D i e V e r e b u n g d e s G e s c h l e c h t s b e i dem
A s k o m y z e t e n Bombard i a l u n a t a Z c k l .
Zechr. indukt.
A b sta m m . u . V e r e r b - l e h r e
7 3 : 1 + 0 3 -9 .

1+0.

Z i c k l e r , H.
1937.
D i e S p e r m a t i e n - b e f r u c h t u n g b e i Bom bar d i a lunata.
B e r . D e u t c h . B o t . G e s . 5 5 : 1 li+—1 1 9~