ATTEMPTS TO INDUCE PARTHENOGLNESIS IN
PLANTS

by
JOSEPH NAMAZI

A THESIS
Submitted to the Graduate School of Michigan State
College of Agriculture & Applied Science in
partial fulfilment of the requirements for the
degree of
MASTER OF SCIENCE
Department of Farm Crops

1951

Ii. twain.)

I.
II.

III.

TABLE OF CONTENTS

Acknowledgement

Introduction . . . . . . . . . . . . . . . . .

Review of literature . . . . . . . . . . . . .

A. Types of parthenogenesis . . . . . . . . .
1. Reduced parthenogenesis . . . . . . .
2. Unreduced parthenogenesis . . . . . .

B. Some examples of parthenogenesis . . . . .
1. Reduced parthenogenesis . . . . . . .
2. Unreduced parthenogenesis . . . . . .

C. Stages in seed setting . . . . . . . . . .

l. The preparation of the medium in which
the seed has to develop . . . . . . .

2. The stages of embryo production . . .

3. Fusion of egg polar nuclei with sperm
nuclei . . . . . . . . . . . . . . . .

Experimental procedure . . . . . . . . . . . .

Experiment I. Hale-sterile sugar beets,
IlaI'Ch, 1950 o o o o o o o o o o o o o o 0

Experiment II. Male-sterile sugar beets,
fall, 1950 o o o I o o o o o o o o o o o 0

Experiment III. Hale-sterile sugar beets,
January, 1951. O O O O O O O O O O O O O 0

Experiment IV. Navy beans, January, 1951.

Experiment V. Trag X Bush beans, spring,
1951 O O O O O O O O O l I O O O O O O O 0

Experiment VI. Determination of presence
of bean pollen in air of the greenhouse .

Experiment VII. Peas, January, 1951 . . .

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18

25
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IV.
V.

Experiment VIII.
Summary and Conclusion

Cited Literature . .

Oats, January, 1951 .

Table
1.

Number of seeds obtained on each sugar beet
plant, Experiment I . . . . . . . . . . . .

Number of sugar beet seed obtained by differ-
ent treatments, Experiment III . . . . . . .

The effect of different treatments upon navy
beans, Experiment IV . . . . . . . . . . . .

The effect of different treatments on bean
flowers, Experiment V . . . . . . . . . . .

The frequencies of red and white hypocotyl
color and of vine and bush types of plants
from flowers of a progeny of Trag and Bush
treated with pea pollen or with vitamin Bl,
Experiment V . . . . . . . . . . . . . . . .

The frequencies of red and white hypocotyl
color and of vine and bush types of plants
from flowers of a progeny of Trag and Bush
allowed to be selfed as control in Experiment

V O O O O O O C O O O O O O O O O O O O O O

16

19

21

23

ACKNOWLEDGEMENT

The writer wishes to express his sincere
appreciation to Dr. E. E. Down, under whose
supervision and encouragement the investigation
on beans was undertaken.

He is indebted to Mr. H. L. Kohls, under
whose guidance and assistance the investigation
on sugar beets was studied.

Gratitude is extended to mr. H..M. Brown
for his willing cooperation in the preparation

of the manuscript.

Attempts to Induce Parthenogenesis in Plants
I. INTRODUCTION

It has long been known that some plants produce
seed by parthenogenesis, the development of an embryo
from an unfertilized egg. The artificial induction
of parthenogenesis by stimulating the stigma of
emasculated flowers by pollinating with foreign pollen
or by dusting with vitamin compounds, chalk or dust is
a comparatively recent development. The study reported
here is concerned with attempted parthenogenesis with
male sterile sugar-beets, common navy beans, peas and
oats.

In the case of plants a desirable character may be
introduced by crossing and back crossing to a plant carry-
ing this character. If parthenogenesis is then induced in
the backcross, we may obtain homozygosity for all characters
without repeated selfing and selection. If haploid plants
are obtained by parthenogenesis, we could get a diploid
homozygous plant by doubling with colchicine. Thus, seed
production by parthenogenesis may be helpful in producing
homozygous plants, which may be used as parent stock for

producing hybrid vigor in cross pollinated plants.

II. REVIEW OF LITERATURE

A. Types of parthenogenesis

According to Sharp (23) we have two types of
parthenogenesis:

1. Reduced Parthenogenesis: In this case the
gamete has the reduced number of chromosomes. This was
called haploid parthenogenesis by Hartman (13) and Renner
(22), and true parthenogenesis by Strasburger (24).

2. Unreduced Parthenogenesis: In this case the
gamete has the unreduced (zygotic) number of chromosomes.
It was called diploid parthenogenesis by Hartman (13) and

Renner (22) and somatic parthenogenesis by Winkler (26).

B. Some examples of parthenogenesis

1. Reduced parthenogenesis has been demonstrated
in datura, tomato, wheat, tobacco, maize, pepper and many
other plants. The first reduced parthenogenesis in plants
was recorded by Blakeslee (1922) (2) in datura and devel-
oped as a sport. Blakeslee and Belling (3) later on ob-
tained a haploid datura plant by crossing Datura firon to
Datura stramonium.

Clausen and Mann (8) described two haploids obtained
by crossing Nicotiana tobaccum X Nicotiana sylvestris.

In the genus Triticum, a haploid plant was found by
Gains and Aase (12). In Triticum compactum, a haploid

developed from a seed formed by pollination with pollen

grains from Aegilops cylindrica. Kihara and Katayama

(16) reported the occurrence of a number of haploid

plants in Triticum monococcum. They induced haploids by
x-raying the young spikes or by merely bagging the spikes
after heading. They also observed the production of
haploids under ordinary cultivation. Katayama (15) x-rayed
the young spikes of Triticum monococcum, estimated to be in
a period of meiosis. Among the treated spikes, haploids
were found at the rate of 5.26 to 7.h1 percent. He also
found haploid plants at the rate of 17.58 percent from the
kernels of Triticum monococcum that were formed by pollina-
tion with x-rayed pollen grains.

Chizaki (6) found a haploid plant in Triticum
monococcum from about 20 seeds that he obtained from retard-
ed spikes that bloomed on a hot day in August.

Nakajima (18) pollinated Triticum turgidum (2 n is 28)
with rye pollen and obtained 182 kernels. Only two of these
germinated and but one of these two was haploid.

Nordenskiold (19) obtained a haploid in rye by heat
treatment of the pollinated flowers. (A A5 minute heat
treatment was given 21 hours after pollination.)

In sorghum haploidy occurs naturally more often than
in most of the Gramineae family and it has been observed
by Brown (A) in the field. The haploid plants can be
recognized since, in comparison with diploids, they are

shorter, less vigorous with more slender stalks, narrower

leaves, smaller and more highly sterile panicles and
smaller glumes.

In rye, Muntzing (17) observed that haploid forma-
tion may be increased by crossing with distantly related
species or by treatment with low temperature at the time
of fertilization. The same effect may also be obtained
by high temperature or x-ray treatment.

Haploids may be naturally obtained in twin seedlings.
Cooper (9) found haploids in twin seedlings of Lilium and
Nicotiana and he believed that one twin arose from the
zygote and was triploid, the other twin from a synergid
nucleus and was haploid.

Haploids in twin seedlings in pepper have been reported
by Christensen and Banford (7).

In recent work, Chase (5) has crossed maize with purple
plumule color (pollen parent) with colorless plumule plant
(female). The progeny obtained by crossing purple plumule
with white plumule (The purple color, Pu, is dominant)
should all be purple, but a few white plumules have also
been found. Those with colorless plumule are tested for
haploidy by morphological characters and by counting the
chromosomes of the root tip. One out of 900 was found to
be haploid among the white and pink plumule seed. The
incidence of haploidy was affected by both parents and,
depending upon parents, varied from Ozh5OO to a high of

1:1h5. In this case the purple plumule pollen acts as an

excitant only.

2. Unreduced parthenogenesis occurs frequently in
vascular plants as a standard reproductive phenomenon
and the resulting plant is not necessarily homozygous,
since it is like the mother plant genetically. Chara
crinita, Marsilea, Thalictrum, Alchemilla, Wikstroemia,
Hieracium, Antennaria and Taraxacum (10) are examples.

The last six are seed plants, the final three being
compositae.

In case of Poa pratensis and Poa palustris (l),
reproduction by parthenogenesis occurs naturally. Arti-
ficial parthenogenesis has been induced in plants such as
in Spirogyra and Chlamydomonas (10) by growing them in six
percent solution of cane sugar. In Marsilea (10) high
temperature seems to be a stimulating factor. Some experi-
ments on artificial parthenogenesis indicate that the pollen
or sperm functions as a growth excitant, rather than a unit
to carry hereditary characters.

In l9h7, Tschermak (25) reported the formation of seed
with parthenogenesis by dusting the carefully emasculated
flower with different compounds in the form of dust, dead
pollen, maizena powder, crushed Betaxin, Cebione tablets,
oat meal, wheat flour, chalk, etc. He repeated the dustings
until the stigma was visually dried. He states that the
seeds that he obtained by stimulating with different com-
pounds are not haploid but rather diploid. The explanation

for diploid parthenogenesis given by Tschermak is quoted

here: "In the case of mere stimulation of development of
the egg-cell, which was reduced to half of its nuclear
sliding composition, the haploidy of the embryo was to be
expected principally, the latter (haploidy) could be regu-
lated to the normal double possession of chromosome by a
single skipping of cell division after nuclear division."
According to fiendel's fundamental law in a single factor
hybrid, the F1 generation has two kinds of gametes A and a
in equal number and when selfed produces a ratio of
1AA:2Aa:1aa and, if A is dominant, a ratio of 3:1 is obtain-
ed. If the F1 is stimulated to produce seed parthenogenet-
ically, we expect the A and a types to develop in equal

numbers giving a ratio of 1:1 homozygous for all characters.

C. Stages in seed setting

Overbeck, Conklin and Blakeslee (20) explain the
process of seed setting and fruit ripening which normally
takes place after pollination in three separate steps.

1. The preparation of the medium in which the seed
has to develop:- This consists in the prevention of
abscission of the fruit and the stimulation of the develop-
ment of the ovary, the placenta, the ovules with their
integuments and, in some cases the nucellus after pollina-
tion but before actual fertilization. This can be shown in
Meilandrium where, after pollination, the ovary and ovules
enlarge considerably before the pollen tube has reached the

embryo sac.

Pavolachko (21) reported that, if the styles of
Nicotiana are removed three hours after pollination, the
pollen caused the development of ovaries and delays drop-
ping.

2. The stage of embryo production:- This stage
takes place after the first step is initiated. It consists
of the division of the polar nucleus and the egg cell and
these initial division are not necessarily connected with
fertilization (in the sense of fusion of nuclei). Ferguson
(11) observed that after pollination of Petunia the polar
nucleus may divide many times before the sperm nuclei have
left the pollen tube. The mere presence of the pollen tube
in the embryo sac seems to be sufficient to cause the
initial division of the endosperm. Jorgensen (1h) observed
that in the cross of Solanum nigrum by Solanum luteum the
sperm nucleus never fuses with egg nucleus but the sperm
nucleus degenerates. Its presence, however, is sufficient
to start division in the egg cell which ultimately develops
into a completely maternal plant. (80 percent of such
plants are diploid while the other 20 percent are haploid.)
The observation of twin embryos indicates that a haploid
embryo can start development under the influence of its
fertilized partner within the same ovule. In no case has
an egg cell been shown to develop under the influence of
the fertilization of an adjacent ovule. This indicates

that, if a diffusable substance stimulates the development

of the egg, its area of activity may be limited. This
might be due to either a large sized molecule which per-
meates membranes with difficulty or to a more readily

diffusible substance with a high rate of destruction.

3. Fusion of egg polar nuclei with sperm nuclei:-
A definite step in typical seed development is the fusion
of the sperm nuclei with egg and polar nuclei which makes
hybridization possible. It has been shown that the nuclear
fusion is neither necessary for growth and enlargement of
ovary and ovules nor for development of the egg into an
embryo. Polyembryony, common in many plants, presents the
best example of embryos formed from somatic maternal tissue.
This occurs however in the presence of an embryo resulting
from fertilization. Sporophyte polyembryony among citrus
shows true embryos which are developed from the nucellus.
A seed of orange regularly contains several viable embryos
only one of which may be the result of fertilization. The
nonsexual seeds will produce seedlings, each with a genetic

constitution like that of the female parents.

III. EXPERIMENTAL PROCEDURE

Experiment I. Male—sterile sugar beets, March 1950.
On March 19, six male-sterile sugar beets (plants
with sterile pollen) were isolated in the greenhouse for
the experiment. Under this circumstance, no crossing or
selfing would be expected. On a portion of a branch, the
stigmas were treated with different chemicals_and labels
were tied at both ends of the treated sections. One of the
beets was untreated. The powders and chemicals were applied
with a camel's-hair bruSh to the stigmas. The time of
application was not regular but an attempt was made to treat
the flowers at different stages from bud to mature flowers.
The treatments were:- 1 - Chalk, 2 - Wheat flour, 3 - Maize
extract, A - Flocer, 5 - Soapstone, 6 - Maize pollen,
7 - Sodium nucleate solution, 1 percent, 8 - Sodium nucleate
solution, 2 percent, 9 - Sodium nucleate powder, 10 - Clip-
ping of the tip branches, ll - Control (nothing applied).
On may 15, the greenhouse temperature rose excessively and
the windows were Opened. This may have led to pollination
of some of the flowers by pollen from plants in a nearby
greenhouse. In order to better analyse the result of this
experiment the seed balls produced were divided into three
classes.
(1) From early flowers that produced seed balls

before the windows were opened.

(2) From mid-early flowers that produced seed balls

-10-

that might have arisen from cross fertilization by pollen
from the nearby greenhouse.

(3) From late flowers that initiated seed balls
after the day when the windows were open.

Three of the male-sterile plants had white flower
buds, the other three had red flower buds. Since some
plants had white hypocotyl and much of the foreign pollen
carried red, the seeds were tested for hypocotyl color.
White-bud plants should have given all white hypocotyl
progeny, if the seeds were produced by parthenogenesis.

But pollination by a mixture of red and white pollen should
have caused segregation into red and white. The test of
hypocotyl color of progenies indicated that pollen from out-
side was the cause of most of the seed formation, as white-
bud beets gave progenies of red as well as of white hypocotyls.
A count of the nearby beets indicated they were a mixed
population in the ratio of 33 white to 77 red beets. Even
when divided into classes on an earliness basis there is a
strong evidence of crossing with nearby beets. Apparently
there was considerable contamination by outside pollination
both before and after the windows were opened, and much
more positive protection from pollen sources must be pro-
vided with sugar beets than is found in a greenhouse con-
taining other beets.

About 15 white hypocotyl plants were selected and root
tip counts of chromosomes were made. All were diploid, 2 n

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- 13 -

From this experiment, the results of which are given
in Table 1, no proper conclusion can be drawn on the

occurrence of parthenogenesis.
Experiment II. Male-sterile sugar beets, fall, 1950.

A few male-sterile annual beets were planted in the
greenhouse in the summer of 1950, where no other beet was
present. During the fall, three plants bolted, but these
plants were weak and because of high temperature most of
their growth was vegetative.

One of the plants was treated with vitamin B1, another
with vitamin C., and one was kept as control. The vitamin
B1 and C. were applied by pepper shaker on the alternate
days until the stigma was dried. This required 6 applica-
tions, at the same time 11 branches, some on each of the
three plants, were bagged and no other treatment applied.

No seed was formed on any of the plants, but it was observ-
ed that by application of these vitamins, in form of powder,
the ovaries were stimulated and were bigger in size, than

untreated ovaries.
Experiment III. Male-sterile sugar beets, January, 1951.

In the winter, 1950, two male-sterile sugar beet
plants were isolated in the Horticulture greenhouse where
there was no possibility of stray beet pollen. The flowers

were treated with different chemicals from bud stage to

-14-

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- 15 -

mature flower and at different times of day. The treat-
ments are listed in Table 2.

The 611 seeds, obtained with the different treatments
shown in Table 2, when planted, did not germinate. Each
seed ball appeared normal but when cut open neither embryo
nor endosperm was present. The hormones caused the forma-
tion of parthenocarpic fruits.

The apple set powder had a very high spreading power
because of its fineness. Due to this cOndition, many of
the seeds that were obtained without treatment are believed

to have been stimulated by the apple set powder.
Experiment IV. Navy beans, January, 1951.

During January, 1951, an attempt was made to produce
seeds in navy beans by parthenogenesis. About 12 navy bean
plants were selected for this experiment. The bean flowers
were emasculated by suction before other treatments were
applied. An eye dropper was fixed to the suction end of a
standard vacuum cleaner. This method of emasculation was
slower than by tweezer, but it had the advantage of not
crushing the stamens and of avoiding accidental pollination.
After this careful emasculation, the flowers were treated

with the substances listed in Table 3.

- 16 -

 

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- 17 _

 

 

 

 

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-18-

The results of Experiment IV shown in Table 3 indicate
that out of 341 emasculated flowers treated with different
substance, 35 pOds were obtained with 98 seeds. However,
the 40 control flowers also produced 7 pods with 22 seeds.

The seeds obtained without pollination were planted
and chromosome counts in root tips were made in few of the
beans. All were diploids. The field test also showed
that they were normal beans.. In this case the navy bean
parents did not have a specific character so that their
segregation ratio could be studied and find whether or not
they would give a ratio of 1:1 according to Tschermak

theory.
Experiment V. Trag X Bush beans, spring, 1951.

According to Mendel's law the first generation (Fl)
of a hybrid, produces two kinds of gametes and two egg cell
and two types of pollen cells (A and a) in equal number.

In further sexual reproduction their union produces a combi-
nation of AA, Aa, aA, aa, in equal number and if one is
dominant a ratio of 3:1 is obtained. But if instead of
sexual reproduction, the gamete produced from F1 partheno-
genesis takes place A and a grow in equal numbers and F2
would be a ratio of 1:1. To find whether the seed produced
in navy beans by stimulation, as in Experiment IV was pro-
duced by parthenogenesis. A cross between Trag and Bush

was made and Eight Fl seeds from such a cross were planted

- 19 -

in the greenhouse in spring of 1951. The flowers on these
eight plants were treated with the different chemicals
shown in Table A.

These parents differed in the following visible,
single factor characters:

Trag Characters Bush Characters

Color of seed Black White
Color of stem Red , Green
Type of growth Vine Bush

The deviations from the Mendilian ratio could be tested

in the F2.

Table A. The effect of different treatments on bean

 

 

 

flowers. Experiment V.
Number

of flowers Empty pods Full pods
No. Treatments treated obtained obtained Seeds
1 Vitamin Bl 193 1 19 6O
2 Pea pollen l6 - 2 12
3 Apple set 19 l - -
A Nothing control 22 - - -
250 2 21 72

 

The 21 pods obtained by pea pollen and vitamin B1
were planted in 1A pots. The two pods treated with pea
pollen had six seeds each and were planted in pots l and 2.

Of those pods from vitamin Bl the pods that had from 6 to 3

- 20 -

Seeds were planted in separate pots (3 to 12); the three
pods with two seeds were planted in one pot (l3); and the
six pods with one seed each were planted in one pot (1A).

The F1 seeds from the cross between Trag and Bush,
according to expectation, produced only red hypocotyl and
vine, as red and vine characters are dominant over white
and bush.

The 72 seeds obtained by stimulation with pea pollen
and vitamin Bl were planted to determine the characters in
the progenies. The results, as to hypocotyl color, from
the treatment of these bean flowers with pea pollen, Table
5, suggest that the seeds were produced by parthenogenesis,
as Tschermak said, but the result as to vine type from these
same flowers do not agree with this. The vitamin Bl treat-
ment, Table 5, gave a 1:1 ratio for vine type, as suggested
by Tschermak, but the result as to hypocotyl color from the
same flowers do not agree with this. The beans that were
not treated and allowed to grow and be selfed normally,
Table 6, gave a ratio of 3 red: 1 white for hypocotyl color,
a ratio of 3 vine: 1 bush for vine type and a ratio of 9 red
vine: 3 red bush: 3 white vine: 1 white bush when the two

characters were considered together.

 

 

 

 

 

 

 

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- 22

 

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- 25 -

Experiment VI. Determination of presence of bean
pollen in the air of the greenhouse, spring, 1951.

To find whether bean pollen was flying in the green-
house eight slides were smeared with vaseline and were
suspended among the bean plants for a month during the
flowering stages of the plants. These slides were exam-
ined under the microscope. Six of these slides did not
have any pollen, one slide had one pollen grain the size
of the bean pollen, and yet another had two pollen grains
of similar size to bean pollen. This shows that very little
bean pollen was flying in the greenhouse. The chance of
pollination was considered very slight especially in view
of the fact that the stigma of bean flower when compared
to slide has very little surface. With this in mind pollen
flying in greenhouse is likely not to have been the cause

of seed formation.
Experiment VII. Peas, January, 1951.

After careful emasculation with tweezers, a few pea
flowers were treated with vitamin B1, vitamin C, apple set
powder and nothing.

The untreated flowers were stimulated only with
emasculation alone and formed six pods, but all the six
pods were empty, the ovary wall was only stimulated.

The following results were obtained.

-26-

The effect of different treatments in pgas.

Number of Empty Full Seeds
No. Treatments flowers treated pods pods obtained

 

 

1 Vitamin Bl 2A 1 2 5
2 Apple set
powder 5 1 - -
Vitamin C 3 - - -
A Nothing
(control) 20 6 _ -
Sums 52 8 2 5

 

Experiment VIII. Oats, January, 1951.

After emasculation with tweezer three heads of oats
were treated with vitamin B1’ few seeds were only stimulated,
that is the ovary was grown to about half the size of normal

seeds but no seed formation took place.

- 27--

SUMMARY AND CONCLUSION

This paper reports an attempt to induce by method
of parthenogenesis the development of pure breeding
homozygous lines of male-sterile sugar beets, navy beans,
peas and oats, by stimulating the stigmas of the flowers
with different compounds following the suggestions made
by Tschermak.

The different substances, in form of dust, stimulated
the growth of seeds in beans and peas, and a few seeds were
harvested without any known fertilization by their own
species. In sugar beets and oats, no seed was obtained.
Those powders that were hygroscopic in nature, like maize
pollen, were less successful on beans than non-hygroscopic
powder. The treatments with different hormones usually
stimulated the growth of ovary wall and not embryo. This
result agreed with the experiment conducted by Overbeck,
Conklin and Blakeslee (20) who treated the Datura flowers
with different concentration of hormones and also injecting
the hormones into young Datura ovaries.

Wherever the seed was obtained by stimulation, Experi-
ment IV, and germinated, the morphological characters of
plants were studied, all the plants tested were normal in
appearance. The root tip counts of a few of the plants
showed only 2 n tissue (diploid).

In Experiment V, involving the progeny of a cross be-

tween Trag and Bush beans, it was possible to study the

-28-

progeny of bean seed obtained by stimulation. With pea
pollen, the progeny showed a 1:1 ratio for hypocotyl color
but a 3:1 for bush : vine. with vitamin B1, just the
opposite was observed.

There is a possibility that in parthenogenesis
unknown conditions may favor the excess development of
some gametes causing an apparent 3:1 ratio, where a 1:1
ratio would be expected. One more generation must be
grown to see whether the populations continued to segre-
gate. If so, it can be concluded that these seeds produced
by stimulation are in the heterozygous condition and this
method is of no value as a short cut to produce homozygous
lines from a Fl hybrid. If further segregation does not
occur, then a shortening of the breeding program will have
been attained and unreduced parthenogenesis will have

occurred.

(l)

(2)

(3)

(A)

(5)

(6)

(7)

(9)

(10)

(ll)

(12)

(13)

- 29 -

V. CITED LITERATURE

Akesberg, E., Seed production of the poa species.
Herb. Rev. 6: 228-223, 1938.

Blakeslee, A. E., J. Belling, M. E. Farnham, and
A. D. Berger, A haploid mutant in the Jimson weed,
Datura stramonium. Science 55: 6A6, 1922.

Blakeslee, A. F., J. Belling, Chromosomal mutations
in the Jimson weed, Datura stramonium. Jour. of
Her. 15: 195, 192A.

Brown, M. S., Haploid plants in sorghum. Jour. of

Her. 3A: 163, 19A3.

Chase, S. S., Monoploid frequencies in a commercial
double-cross hybrid maize and its component single
cross hybrid and inbred lines. Genetics 3A: 328,
19h9.

Chizaki, Y., Original not seen, cited by
Nordenskiold (16).

Christensen, H. C. and R. Banford, Haploid in twin
seedlings of pepper. Jour. of Her. 3A: 99, 19A3.

Clausen, R. E. and M. C. Mann, Inheritance in
Nicotiana tobaccum V. The occurrence of haploid
plants in interspecific progenies. Proc. Nat. Acad.
Science 10: 121, 192A.

Cooper, D. 0., Haploid, Diploid twin embryos in
Lilium and Nicotiana. Amer. Jour. of Bot. 30: A08,
1943.

Coulter, J. E., C. R. Barnes, H. C. Cowles, A text
book of Botany, volume 3, Ecology, pp. 398. New
York: American Book Company, 1931.

Ferguson, M. 0., A cytological and genetical study
of petunia. Torrey Botanical Club. 5A: 657-66A,
1927. -

Gains, E. F. and H. C. Aase, A haploid wheat plant.
Amer. Jour. of Bot. 13: 373, 1926.

Hartmann, Original not seen, cited by Sharp (23)

(1A)

(15)
(16)
(17)
(18)
(19)

(20)

(21)

(22)
(23)

(ZA)
(25)

(26')

- 3o -

Jorgensen, C. A., The experimental formation of
heteroploid plants in the genus solanum. Jour.
of Gen. 19: 136, 1928.

Katayama, Y.,
Triticum monococcum.

Haploid formation by x-ray in
Cytologia 5: 235, 192A.

Kihara, H. Y. Katayama, Original not seen, cited
by Katayama (15).

Huntzing, A., Note on a haploid rye plant.
Hereditas 23: A0, 1937.

Nakajima, G., Occurrence of a haploid in Triticum
turgidum. Jap. Jour. of Genetics 11: 2A6, 1935.

Nordenskiold, H.,‘ Studies of haploid rye plant.
Hereditas 25: 20A, 1939.

Overbeck, J. V., M. E. Conklin and A. F. Blakeslee,
Chemical stimulation of ovule development and its
possible relation to parthenogenesis., 28: 67A, l9Al.

Pavolachko, P. A., Original not seen, cited by
Overbeck (20).- '

Renner, Original not seen, cited by Sharp (23).
Sharp, L. E., Introduction to Cytology, pp. A02.
New York and London, NcGraw-Hill Book Company,
193A.
Strasburger, Original not seen, cited by Sharp (23).
Tschermak - Seysenegg, E. Vienna. Artificially
effected seed formation without fertilization.
Received November 20, 19A7.

Winkler, Original not seen, cited by Sharp (23).

 

 

 

 

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