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"Studies on the Resistance of Two
strains of Navy (Pea) Beans to

. " Virus 1550”

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STUDIES ON THE RESISTANCE OF TWO S'I'RADIS

OF NAVY(PEA) BEANS TO VIRIB 15

by
HAE‘mOND PERCIVAL FORD

“I”

A THESIS
Submitted to the School of Graduate Studies of Michigan
State College of Agriculture and Applied Science
in partial fulfillment of the requirements
for the degree or

MASTER OF SCIENCE

Department of Fan Crepe
1950

WHEg;

STUDIES ON THE RESISTANCE OF TWO SPRAINS

01' NAVNPEA) m TO name 15

QRAAQQ

ACMOWIEDGEWIH
The author wishes to acknowledge his indebtedneee to Dr. E. E.
Down, Profeeeor or and Research Professor in Fan Crops, for 111.
guidance and aid in making this investigation.
The author is grateful to Dr. A. I... Anderson, Pathologist in
Bureau of Plant Industry U.S.D.A. for hie assistance and «Operation

in pathological work in this investigation.

I.

III.

V.

Introduction and review of literature . . . . . . . . . .
:MBthOdB and.mater1ale e e e e e e e e e e e e e e e e e e

Presentation and discussion or data . . . . . . . . . . .

A.
1.
2.
3.
4.
5.

6.

Summary e e e e e e e e e e e e e e e e e e e e e e e e

Literatur. Citede e e e e e e e e e e e e e e e e e e e

Tables

Table

Table

Table

Table

Table

Table

1 . .

2 and

TABLE OF CONTENTS

P380
1

11

13
15
16

18

INTRODUCTION

Robust been, a pea bean variety was introduced into Michigan in 1914
by F. A. Spragg. (1)1

The Robust variety was developed from a thrifty selection made in
1907. Reddick reported in 1918 that this new variety was resistant to can-
non bean mosaic now called virus 1. (1)

It is probably because of this one characteristic more than any other
that the Robust variety was used to plant 85 percent of the Michigan pea
bean acerage in 1938. (2)

The Robust variety was used as one of the parents of the Michelite
been, a pea been that was introduced into Michigan in 1938 by the Michi-
gan Agricultural Experiment Station. (2)

Reports from New York State in 1939 showed Michelite to be suscept-
ible to a new virus. (3) This new virus has been called strain 15, a
variant of strain 1 (4), Burkholders strain (5). In the work done here,
it has been called strain 15.

The acreage of Michelite beans increased in Michigan until in 1948
when an estimated 400,000 of the 500 ,000 acres of pea beans were planted
to it. Virus 15 was first observed by A. L. Anddrson in Michigan in 1948.
(Unpublished)

Because Michelite and Robust varieties are susceptible to virus 15
and all other varieties of pea beans grown in the state are susceptible to
virus 1 and virus 15, the spread of virus 15 could do a great mount of
damage. Consequently a breeding program was set up by the Michigan Agri-

cultural Experiment Staticn in cooperation with the Division of fruit,

 

lNumbers in parentheses refer to literature cited.

(1)

 

 

.. .-

 

vegetable crepe and plant diseases of the Bureau of Plant Industry of the
U.S.D.At in 1948.

As a source of resistance to virus 15, a strain of beans number 45-62
selected(by H. M. Munger of Cornell University, Ithaca, New York) from the
354 generation of a back cross between Michelite I Great Northern 31 re-
crossed to Michelite was used. This strain has many desirable characteris-
tics, but it has a badly wrinkled seed coat when grown in the field. This
fault prevents it from being introduced as a commercial variety. 'Mdchelite
and selections from several crosses between Robust and Crawford and other
varieties were crossed with selection 46-62. It was observed that in the
cross between selection number 50021 and 46-62 susceptible and resistant
FBI plants were produced. Immediately, the purity and value of strain 46-62
for virus 15 resistance was questioned.

The present investigation.wes undertaken to determine the inheritance
of virus 15 resistance as found in strain number 48-62.

Ali in 1950 (6) reported on the genetics of resistance to been virus
1. The results from this indicated that there was dominant resistance
present in Corbett Refugee type of beans, and a recessive factor for re-
sistance present in the Robust varieties. His data gave evidence of two
pairs of independently inherited gene differences, with.deminant and re-
cessive epistasis.

In.werk:reported in 1929(7) from.the Expertment Station of the Uni-
versity of Idaho, it was found that at low temperatures the mosaic symp-
toms of been virus 1 were reduced in severity, and that there was eunuch
lower percentage of infection of susceptible plants. Susceptible plants
were inoculated with virus 1 and placed in chambers at three different 1e-

vels of temperature. .At the end of twenty days, the plants held at the two

(2)

higher temperatures (60 and '70 degrees Fahrenheit) showed a considerable

amount of mosaic, while those kapt at a twperature of 4? degrees Fahren-
heit did not show any mosaic symptoms. The plants which had been kept

at the low temperatures were then placed in a chamber which had a temper-
ature of ‘70 degrees Fahrenheit. At the end of two weeks, five of the ten
inoculated plants showed mosaic symptoms.

In 1948, Grogan and Walker (5) reported on the effect of temperature
on necrosis found on plants which were inoculated with the approach graft
method and subjected to different temperatures. They used temperatures
of 82, 75, 68, and 60 degrees Fahrenheit. Three strains of viruses that
attach beans were used. They were the comes mosaic (strain 1) greasy pod,
and strain 15. Fourteen days after inoculation many plants inept at 82 and
75 degrees were dead and a few of the plants kept at 68 degrees and inocu-
lated with virus 1 were dead while those inoculated with the greasy pod
strain and strain 15 were Just beginning to show necrosis at this temp-
erature. In plants kept at 60 degrees, there were a few plants that had
been inoculated with virus 1 and with the greasy pod strain which were
showing necrosis, but there was no necrosis in the plants inoculated with
virus 15. After 28 days practically all of the plants that had showed
necrosis earlier were dead in the chambers where the tauperatures were
maintained at 82, 75, and 68 degrees. Plants which were inoculated with
virus 1 and greasy pod strain and maintained at 60 degrees were showing
a high percentage of necrosis but no plants were dead. Plants which had
been inoculated with virus 15 and kept at 60 degrees showed a slight
mettle on one variety while on the second variety there were no symp-
taas. When the plants which showed no symptoms were moved to a house

with a temperature of 82 degrees, they ddveloped necrosis after five days.

(3)

MATLRIAIS AND METHODS

Two strains of navy pea beans were used in this investigation.
One strain, which was susceptible to virus 15, was selection number
50021 from the cross between the Robust and the Crawford pea beans. The
other strain was selection number 46-62 made by H. M. Munger of Cornell
University, Ithaca,New York. This strain originated from a cross between
the'Michelite pea bean and Great Northern 31. iMichelite is susceptible
to virus 15 and has the Robust variety as one of its parents, Great Nor-
thorn 51 is a Great Northern type of bean and is resistant to virus 15.
The F51 plants were back crossed thMichelite for three generations with
the resistant plants being selected. Strain 46-62 is resistant to virus
15 and is nine days earlier than the Michelite, but it has a badly wrinkled
seed coat when grown in the field. This fault prevents its introduction
as a commercial variety.

The strain of virus 15 which was used was one that was obtained from
W. J. Zhumeyer of the UtS.D.A. It was carried on susceptible plants in
the green house and had been used in previous work in this progrsla

All the crossing and the testing for disease resistance was done in
the greenhouse. The temperatures in the green house at the time of ino-
culation varied to a considerable extent with the outside temperatures.
The temperature was fairly high when the plants were inoculated in the
early fall while the temperatures were fairly low when the plants were
inoculated in early January. The temperatures were observed to be down to
about 50 degrees Fahrenheit in the mornings during January. However there
were no daily temperature recordings made. The greenhouse was kept almost

free of insects by frequent fumigations.

(4)

Cross pollination was carried out in two different ways. In method
one the anthers were removed by the use of forceps from the flower that
was to be used as the female parent. Then a stiwa was taken from a
flower that had already pollinated and which was to be used as the male
parent. (2) This stigna was used to pollinate the stigna of the female
flower. In method two the emasculation of the flower to be used as the
female parent was done through the use of suction. The vacuun was sup-
plied by an ordinary tank type vacuum sweeper from which a rubber tube
was extended. This tube had a pointed glass tip fitted in the end.

The tip was used to draw off the anthers one at a time. Pollination was
done in the same manner as in method one. The results from this method
were as good as from using the forceps and it was faster.

The plants were inoculated with the virus in two ways. In method
one known as the rub method (2) the carborundum was Spread on the prim-
ary leaf and then the leaf was rubbed with a pad of cheese cloth that
had been saturated in a suspension of virus 15. The suspension of virus
15 was made by extracting the Juice fraa diseased leaves. This Juice
was diluted with distilled water at a ratio of five parts water to one
part juice. Method two consisted of spraying the primary leaves with the
suspension of the virus into which carborundum had been placed. The
Juice was diluted with nine parts of water to one part of Juice. The plants

were all inoculated when they reached the primary leave stage.

(5)

DISCUSSION

Both susceptible and resistant plants were found in the FBI gen-
eration of crosses between strains 50021 and 46-62. Therefore, work
was started to determine if there were any differences in reaction of dif-
ferent plant selections of strain 46-62. Selections of strain 46-62
reacted differently when inoculated with virus 15. In inoculating a
second generation of these same selections, they did not always react
in the same.manner. This indicated that the differences.might be due
to invironmental conditions rather than a difference in genetic con-
stitution. Results obtained when strains of 46-62 used as a female
were crossed with strain 50021 are shown in table 1. All of the select-
ions gave similar results in the Fbl and F52 generations regardless of
the reaction of the parent selection. This indicates that strain 46-62
is homozygous for resistance and the difference in reaction of indiv-
idual plants to inoculation with virus 15 is due to environmental factors.

.All of the 3-1 generation of plants was susceptible to virus 15
when they were inoculated in the fall.(Tab1e 2) In the crosses where
strain 50021 was used as the female, the results were practically the
seme.(Tab1e 5) This would indicate that resistance to virus 15 was
governed by a recessive factor or factors. The Fbl generation of plants
which was inoculated in the winter segregated into a ratie of about five
resistant plants to one susceptible.(Table 4) In the crosses where
strain 50021 was used as the female, the F31 plants segregated into
a a ratio of 23 resistant plants to one susceptible(teble 5), indi-
cating that resistance to virus 15 was governed by a dominant factor or
factors.

The 332 generation which was inoculated in the winter was fron.the

FBI generation that had been inoculated in the fall. This F-z seg-

(a)

Table 1
Shows the reactions of the F-1 and F-2 generation to virus 15 from
crosses between susceptible strain 50021 and different selections of re-
sistant strain 46-62 as the female.

Section 1 selections showing no reaction to inoculation.
(a) F-l plants inoculated in fall and F-2 plants inoculated in winter.

F-2 F-z
F51 generation generation
Selection generation F-l resistant F-l susceptible
number Res. Sus. Res. Sue. Res. Bus.
02 0 2 28 12
10 .9. .52.. .19. .11
Total 0 5 46 23

(b) Fhl plants inoculated in winter Fk2 plants inoculated in spring.

02 1 o a 9
1o 3 5 e 13 _9__ 26
total 4 5 11 22 9 6

Section 2 selections showing vein browning on the lower leaves when in-
oculated.
(a) F-l plants inoculated in winter F-2 plants inoculated in the spring.

05 l 1 1 3 2 9
06 11 1 21 36 2 10
14 0 1 no plants

19 .9. .2. .15.. as. .2. _9_
total 17 5 38 85 6 28

Section 3 selections showing necrotic flecking of stem and petiole when
inoculated.
(a) F-l plants inoculated in fall F-z plants inoculated in winter.

o8 o e 90 24
09 _9__ 4 52 18
total 0 12 152 42

(b) F-l plants inoculated in winter F52 plants inoculated in spring.

03 5 0 13 36

04 3 0 9 25

08 l l 2 10 0 5
09 19 2 60 140 4 8
11 7 0 23 61

12 4 l 13 36 no plants

15 3 0 12 24

1'7 .2. .1. .11: .32 _ __
total 45 5 145 357 4 l3

('7)

Table 1 (Continued)

Section 4 selections showing lower leaf necrosis when inoculated.

(a) F51

Selection
number

01

13

16

total

plants inoculated in winter FLZ plants inoculated in spring.

3&2 F32
generation generation
generation FBI resistant F51 susceptible
Res. Sue. Res. Sue. Res. Sue.
10 l 16 87 3 8
10 1 25 79 no plants
.17. .1. _.1_3. _§.§. .32. plants
27 3 54 201 3 8

(8)

Table 2

Shows the reactions of the F51 and F-2 generations to virus 15
from crosses between susceptible strain 50021 and different selections
of resistant strain 46-62 as the female when the inoculation of the

F31 was made in the fall and the 3-2 was inoculated in winter.

rue

F-l generation
Selection generation 351 plants sue.
N031“: Rfl.e 811's RBBe We
02 0 2 28 12
08 0 8 90 24
09 0 4 62 18
10 .9. .2. .19.. .11
Total 0 17 198 65

F’Z segregation. Chi .quare(3-1) 00114 Po range 0.70-0.80

Table 3

Shows the reactions of the F-1 and F52 generations to virus 15
from.crosses between susceptible strain 50021 and different selections
of resistant strain 46-62 as the.male~when the inoculation of the 351

was.made in the fall and F52 was inoculated in winter. No selection

numbers used. “J
1&1 3-2
generation generation
Res. Sus. Res. Bus.
1 8 78 34
0 5 43 9
1 7 50 -J§§
Total 2 20 171 106

(9)

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regated into a ratio of approximately three resistant plants to one
susceptible.(Table 2) In the 332 generation where strain 50021 was
used as the female in the F51, the ratio was less than two resistant
plants to one susceptible.(Table 3) Again this would indicate that
the resistance to virus 15 was dominant and due to one pair of factors.
The F-2 generation which was inoculated in the spring was from.the F51
plants that were inoculated in the winter. This F-2 population seg-
regated into a ratio of approximately one resistant plant to three
susceptible.(Table 4) When strain 50021 was used as the female in the
331 generation, the F52 pepulation segregated into a ratio of approxb
imately one resistant plant to three susceptible.(Table 5) These
results indicate again that resistance to virus 15 was recessive and
due to a one factor difference.

The back cross populations are small but they verify the results
obtained with the Fhl and F52 populations. The BC-l generation in.
which the F51 plants were used as the female and strain 50021 as the male
was inoculated in the winter. The BO-l population segregated into a
ratio of approximately one resistant plant to one susceptible.(Tab1e 6)
This would indicate a dominant type of resistance to virus 15 and due
to one pair of factors.

The BC-l generation in which strain 50021 was used as the female
and FBI plants as the.male was inoculated in the spring. This gener-
ation was almost all susceptihle to virus 15.(Table 1) This would ind-
icate a recessive type of resistance to virus 15.

The difference in results in.which dominant and recessive resis-
tance is indicated is explained by incomplete penetranoe of a single
recessive factor pair. This is when the heterozygote is influenced by

environmental conditions. That is it.may be resistant or susceptible

(10)

Table 4

Shows the reactions of the F31 and F52 generations to virus 15 from
crosses between susceptible strain 50021 and different selections of re-
sistant strain 46-62 as the female when the inoculation of the 351 plants

was.made in winter and the F-2 plants were inoculated in spring.

F-z 322
3-1 generation generation

Selection generation 151 resistant Fbl susceptible
number Res. Sus. Res. Sus. Res. Bus.
01 10 1 16 87 3 8
02 1 0 3 9
03 5 0 13 36
04 3 0 9 25
05 1 1 l 3 2 9
06 11 1 21 36 2 10
08 l 1 2 10 0 5
09 19 2 60 140 4 8
10 3 5 8 l3 9 26
11 7 0 23 61
12 4 1 13 36 no plants
13 10 1 25 79 no plants
15 6 0 12 24
16 7 1 13 35 no plants
17 3 1 13 25 2 7
19 .5. .2. .15. .29. 3... .2
total 89 17 248 665 24 77

F52 segregation Chi square(3:1) 2.57 In range 0.10-0.20

(11)

Table 5

Shows the reactions of the F-1 and F-2 generations to virus 15 from
crosses between susceptible strain 50021 and defferent selections of re-
sistant strain 46-62 as the male when the inoculation of the FLl‘was.made

in the winter and of the 352 plants in the spring.

F52 The
F-l generation generation

Selection generation F-l resistant F-l Susceptible
Dumbo: Q: “1. m. a Bu. 0 R’. e Sui R98 0 We
01 7 0 20 65
04 5 O 14 34
06 7 1 29 57 no plants
12 9 0 29 78
14 6 0 18 53
15 8 0 25 73
17 .2. .1. .13. .31 no plants
total 46 22 147 397

(12)

Table 6

Shows the reaction of the BC-l generation to virus 15 when sus-
ceptible strain 50021 was used as the male and the 351 plants were
used as the female. No selection numbers were used. Inoculations
were made in the winter.

BC-l

generation
mfle Sue

fill-‘GOHO
OI'HOGOH

Total

(1:5)

depending on the eswironment. When.the temperatures are cool, the
heterozygote is resistant to virus 15, and when the temperatures are
hot, the heterozygote is susceptible. So the results given would
indicate that resistance to virus 15 in strain 46-62 is due to a rec-
essive single factor pair of genes. This is shown by the ratio-of one
resistant to three susceptible plants in the F52 generation, and by
the all susceptible BC-l generation, both of which were inoculated in
the spring. Also by the F51 generation which was susceptible when
inoculated in the fall. The ratio of three resistant plants to one
susceptible and the ratio of one resistant plant to one susceptible in
the 30-1 generation both of which were inoculated in.the winter can be
explained by the cool temperatures causing the heterozygote to be
resistant.

These data establish the fact that resistance to virus 15 in strain
46-62 is governed by a single recessive factor, that expresses itself
at low temperatures as three resistant plants to one susceptible and

at high temperatures as one resistant plant to three susceptible.

(14)

T®h7

Shows the reactions of the Bc-l generation to virus 15 when sus-
ceptible strain 50021 was used as the female and the F51 plants were
used as the.ma1e. Inoculations were made in the spring. The number of
the selection of resistant strain 46-62 used in.making the cross for

the F-1 generation is given under the selection number.

we
Selection generation
number Sue.
m
m
M
M
w
m
m
w
m
n
R
M
H
tad

HH

mkooowoowoomwo?
D
C

q H

okuooooomooaaa

(15)

SUMMARY

The Michelite variety of pea'beans, which is used to plant most
of the pea bean acreage in Michigan, is susceptible to virus 15 a
new virus disease that infects beans. Therefore, a breeding program
was started to incorporate resistance to virus 15 into the Michelite
variety. As a source of resistance strain 46-62, from.Corne11 Un-
iversity, Ithaca, New York, was used. When this strain was crossed
with several other strains of susceptible beans, both susceptible and
resistant F51 plants were produced. Therefore, this investigation to
determine the genetic inheritance to resistance to virus 15 as found
in strain 46-62 was carried out. Resistant strain 46-62 was crossed
with susceptible strain 50021 to study the genetic inheritance.

The FBI plants produced from this cross were all susceptible
when they were inoculated in the fall, indicating that resistance was
governed by'a recessive factor or factors. .Another E-l generation was
inoculated in the winter and these plants segregated into a ratie of
approximately five resistant plants to one susceptible, indicating
a dominant type of resistance.

The r-z generation of the F-1 plants which were inoculated in
the fall segregated into a ratio of approximately three resistant
plants to one susceptible, indicating that resistance was governed by
a dominant single factor pair. The F52 generation of the 331 plants
which were inoculated in the winter segregated into a ratio of ap-
proximately one resistant plant tc three susceptible, indicating that
resistance was governed by a recessive single factor pair.

The BC-l generations which.were inoculated in the winter and in
the spring were small but they verify the other findings by indi-
cating dominant and recessive factors in the winter and spring re-

spectively.

hm

This variance in results can be explained by the effect of tem-
perature on the heterozygote. The recessive gene exhibits incomplete
penetrance. When the temperatures are low the heterozygote is re-
sistant to virus 15, but when the temperatures are high the heter-
ozygote is susceptible to virus 15.

These data establish the fact that resistance to virus 15 in
strain 46-62 is governed by a single factor pair of genes. Resis-
tance is expressed in the heterozygote as a recessive gene when the
temperatures are high and as a dominant gene when the temperatures are

low.

(17)

(1)

(2)

(3).

(4)

(5)

(6)

('7)

LITERATURE CITED

Spragg, F. A. and Down, 1:. E. The Robust bean. Michigan State
College Agricultural Experiment Station section of Farm Crops
Special Bulletin 108. 1921

Down, E. E., and Thayer, J. W. The Michelite bean. Michigan
State College Agricultural Experiment Station section of
Fan Crops Special Bulletin 295. 1938

Richards, B. L., and Burkholder, W. H. A new mosaic disease
of beans. Phytopathology 33:1215-1216. 1943

Zaumeyer, W. J., and Barter, L. L. Two new virus diseases of beans.
Journal of Agricultural Research 67:305-328. 1943

Grogan, R. G. , and Walker, G. C. The relation of common mosaic
to black root of been. Journal of Agricultural research 77:
315-331. 1948

Ali, M. A., Genetics of resistance control to the canon bean
mosaic virus(bean virus 1) in the bean, Phytopathology 1:66-
79. 1950

Pierce, W. H., and Hungerford, C. W. Symptomatology, transmission,

infection and control of bean mosaic in Idaho. University of
Idaho Agricultural Experiment Station Research Bulletin 7. 1929

(18)

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