Male sterility in cotton

PRESENT PERSPECTIVE OF HYBRID SEED
PRODUCTION USING MALE STERILITY IN COTTON
Mr. Pansuriya Yashkumar A.
M.Sc (Agri.) Student, Genetics and Plant Breeding
Reg. No. :- 2010117084
Subject :- GP 591
Time :- 8:00 to 9:00
Date :- 20 October 2018
Minor Guide:
Dr. J. B. Patel
Associate Professor
Dept. of Seed Science & Technology
COA, J.A.U., Junagadh
1
Major Guide :
Dr. M. G. Valu
Associate Research Scientist
Cotton Research Station
J.A.U., Junagadh

Introduction
Hybrid seed production methods
Mechanism of male sterility
Male sterility genes are identified in
cotton
Review of literature
Conclusion
Contents
2

Introduction
 Karpas/Karpasi - Sanskrit
 Karposas –Greek, Carbasus- Latin
 Cutan – Arbi Cotton – English
 Key role in human civilization
 Vital role in national economy
 White gold
 Dominancy over synthetic fibers
 Tree of wool
Subkingdom -
Tracheobionta
Superdivision -
Spermatophyta
Division -
Magnoliophyta
Class - Magnoliopsida
Subclass - Dilleniidae
Order - Malvales
Family - Malvaceae
Kingdom - Plantae
Vascular Plant
Seeded Plant
Flowering Plant
Dicot Plant
Mallow plants
Genus - Gossypium
4Source: https://en.wikipedia.org/wiki/Cotton

Introduction
 Cotton, ‘King of fibre’ is the premier cash crop in India.
 It is cultivated in tropical and subtropical regions of more than
80 countries of the world.
 India is the pioneer country for the cultivation of cotton
hybrids on commercial scale after releasing of Hy-4
developed by Dr. C. T. Patel in 1971 from Main Cotton
Research Station, Surat.
 Play a vital role in agriculture, industry, social and monetary
affairs of country’s economy.
 More than 150 cotton hybrids have been released by various
State Agricultural Universities and private seed companies.
4

Species Chro.
number
Origin Genome Genome
size
Old world cottons
G. herbaceum 2n=26 Africa A1 1700 Mb
G. arboreum 2n=26 Indo-Burma A2 1746 Mb
New world cottons
G. hirsutum 2n=52 Central
America
AD1 2500 Mb
G.barbadense 2n=52 South
America
AD2 2500 Mb
Cultivated species of cotton
5Source: http://agropedia.iitk.ac.in/content/cultivated-
species-cotton

6
No. Species Genome Distribution No. Species Genome Distribution
Diploid (2n = 26) 26 G. thurberi D1 America
1 G. africanum A Africa 27 G. armourianum D2-1 America
2 G. herbaceum (Cult.) A1 Afghanistan 28 G. harknessii D2-2 America
3 G. arboreum (Cult.) A2 Indo-Burma 29 G. klotzschianum D3-k America
4 G. anomalum B1 Africa 30 G. davidsonii D3-d America
5 G. triphyllum B2 Africa 31 G. aridum D4 America
6 G. barbosanum B3 Cape Verede 32 G. raimondii D5 America
7 G. capitis-viridis B4 Cape Verede 33 G. gossypioides D6 America
8 G. sturtianum C1 Australia 34 G. lobatum D7 America
9 G. nandewarense C1-n Australia 35 G. trilobum D8 America
10 G. robinsonii C2 Australia 36 G. laxum D9 America
11 G. australe C3 Australia 37 G. turneri “D” America
12 G. pilosum K Australia 38 G. stocksii E1 Arabia
13 G. costulatum K Australia 39 G. somalense E2 Arabia
14 G. populifolium K Australia 40 G. areysianum E3 Arabia
15 G. cunninghamii K Australia 41 G. incanum E4 Arabia
16 G. pulchellum K Australia 42 G. longicalyx F1 Africa
17 G. nelsonii K Australia 43 G. bickii G1 Australia
18 G. enthyle K Australia Allotetraploid (2n = 52)
19 G. londonderriense K Australia 44 G. hirsutum (Cult.) (AD)1 America
20 G. marchantii K Australia 45 G. barbadense (Cult.) (AD)2 America
21 G. exiguum K Australia 46 G. tomentosum (AD)3 Hawai
22 G. rotundifolium K Australia 47 G. lanceolatum (AD) America
23 G. fryxellii K Australia 48 G. mustelinum (AD) America
24 G. binatum K Australia 49 G. darwinii (AD) America
25 G. nobile K Australia 50 G. caicoense (AD) America
07Source: CICR Tech. Bull. No. 5
Table 1: Gossypium species, their genomes and distribution

Table 2.Cotton area (lakh ha), production (lakh bales of 170 kg) and
productivity (kg/ha).
State Area ( lakh ha) Production(lakh bales)
Productivity
(kg/ha)
Maharashtra 41.98 85.09 344
Gujarat 26.18 104.1 675
Telangana 18.24 57.35 531
Karnataka 5.65 19.55 572
Andhra pradesh 5.44 22.97 688
Harayana 6.56 25.22 648
Madhya pradesh 5.99 20.3 568
Punjab 3.85 12.5 529
Rajasthan 5.03 22.9 544
Tamil Nadu 1.48 6.80 689
Odisa 1.45 3.95 351
Others 0.50 2.00
India 122.35 377 524
World 333.85 1213.72 798
AICCIPANNUAL REPORT: 2017-2018
7

Hybrid seed.........
 In India, hybrids cover about 45% of total cotton
area and contribute about 55% of the country’s
cotton production.
 Preference due to hybrid vigour.
 But, seed is very costly ranging from Rs. 600/- to
Rs. 1000/- per kg, which cannot be afforded by
small and marginal farmers.
8
Source: http://www.agbioforum.org/v12n2/v12n2a03-
sadashivappa.htm

Hand emasculation
and pollination
Hybrid seed production methods
9
Male sterility based
hybrid
Source: https://link.springer.com/chapter/10.1007/978-3-319-27096-8_4

1. Hand emasculation and pollination method
 Development of such hybrids involve three steps
viz.,
I. Identification and growing of male and female
parents
II. Emasculation of female parent
III. Pollination of female parent with identified male
parent.
 Majority of the hybrids released by hand
emasculation and pollination method.
10

2. Male sterility based hybrid method
 Development of such hybrids involve two steps viz.,
1. Identification and growing of male and female parents
(GMS / CGMS )
2. Pollination of female parent with identified male (selected /
R line) parent.
 Bypass the process of emasculation.
11

Mechanism of Male Sterility
12

What is male sterility ?
Male sterility refers to a condition in
which nonfunctional pollen grains
are produced in flowering plants.
 In flowering plants, the first case of
male sterility was reported by
Koelreuter in 1763.
In cotton, the first case of male
sterility was reported by Justus and
Leinweber in 1960 in upland cotton
(Gossypium hirsutum L.).
13

Phenotypic expression classes of male sterility
I. Structural male sterility : anomalies in male sex organs (or
missing all together)
II. Sporogenous male sterility : stamens form, but pollen
absent or rare due to microsporogenous cell abortion before,
during, or after meiosis
III. Functional male sterility : viable pollen form, but barrier
prevents fertilization (anther indehiscence, no exine
formation, inability of pollen to migrate to stigma e.g.
soybean, peas )
14Source:https://www.researchgate.net/
Heterosis_Breeding_in_cotton

Defective pollen wall contributes to male sterility in cotton
 To understand the mechanisms of male sterility in cotton
(Gossypium spp.), combined histological, biochemical and
transcription analysis using RNA-Seq was carried out in the
anther of the single-gene recessive genic male sterility system
of male sterile A line and male fertile B line, which are near-
isogenic lines (NILs) differing only in the fertility trait.
 Cluster analysis and functional assignment of differentially
expressed genes revealed differences in transcription
associated with pollen wall and anther development, including
the metabolism of fatty acids, glucose, pectin and cellulose.
15China Wu et al. (2015)

Conti……
 In case of CMS, tapetum abnormality is found to be the cause
of sterility. The microsporogenesis in sterile and fertile lines is
similar in early stages but in the later stages the tapetum in the
anther sac of sterile lines disintegrated at pollen mother cell
stage.
 Physiological and biochemical analysis indicates that starch
levels are lower in male sterile plants throughout the entire
period of pollen development, and the soluble sugar level in
young stamen of sterile plants are higher than in fertile plants.
16China Wu et al. (2015)

• (A) Male fertile plant (left) and male sterile plant (right) at full-bloom stage.
• (B) Male fertile flower (left) and male sterile flower (right), with petals
removed.
• (C) In a male fertile anther (left), dehiscence is normal; while in male sterile
anther (right),dehiscence is abnormal.
• (D) Male fertile pollen grains stained with 1% KI solution showing mature
pollen grains that are dyed black.
• (E)While male sterile pollen are not stained.
Fig.1 : Phenotypic comparison between the male fertile and male sterile lines.
16

• (F–K) Locules from the anther section of the male fertile (F–H) and male sterile
(I–K) plants
• There are no differences between the (F) and (I) plants at particular stage
• Compared to those of the plant (G), the spines could not be identified on the
surface of pollen in the plant (J).
• The microspore cytoplasm was stained deeply in the plants (H), but the
microspores aborted in plants (K).
Fig.2 : Phenotypic comparison between the male fertile and male sterile lines
anther cross section
17

Classification of male sterility
1) Genetic male sterility (GMS)
2) Cytoplasmic male sterility (CMS)
3) Cytoplasmic genetic male sterility (CGMS)
19

1. Genetic male sterility (GMS)
 The pollen sterility that is
caused by nuclear genes is
termed as genic or genetic
male sterility.
 In cotton, GMS has been
reported in upland, Egyptian
and arboreum cottons.
 In tetraploid cotton, male
sterility is governed by both
recessive and dominant genes.
20Bulletin no: 24 Anon., 2002

Types of genetic male sterility (GMS)
I. Environment insensitive GMS : ‘ms’ gene expression is
much less affected by the environment.
II. Environment sensitive GMS : ‘ms’ gene expression occurs
within a specified range of temperature and/or photoperiod
regimes i.e.
A. TGMS
B. PGMS
C. Transgenic male sterility
21
Source:https://www.researchgate.net/

A. Thermo-sensitive genetic male sterility (TGMS)
 Sterility is at a particular temperature.
 TGMS system in G. arboreum has been identified for the
first time.
 The line remains sterile till temperature reaches 24ºC and
show complete pollen fertility at temperature less than
18°C.
22

B. Photoperiod-sensitive genetic male sterility (PGMS)
 Sterility is obtained in long day conditions while in short
day normal fertile plant.
 PGMS system in G. hirsutum has been identified for the
first time.
 PGMS lines in G. hirsutum show complete pollen sterility
when temperature rises above 40 ºC for continuous period
of time.
23

TGMS & PGMS system in cotton
 TGMS is a two line method of hybrid seed production where the
sterile flowers are converted into fertile as well as fertile into
sterile at a particular temperature. Once this is stabilized it is
possible to convert any elite variety into TGMS and develop
heterotic hybrids.
 The period in between the completely fertile and completely sterile
phase which produced partially fertile /partially sterile flowers were
observed to be the sensitive stage.
 The flower behavior during normal growing season in the field
condition was observed continuously in this line for four
consecutive years (2007, 2008, 2009 and 2010)
25CICR, Nagpur Palve et al. (2011)

Sterile Partially Fertile Completely Fertile
Fig.3: Sterile, partially fertile and complete fertile flowers of TGMS line

Conti……..
 During the years 2007, 2008 and 2009, the flowers produced at the
onset (i.e. during last week of August to first week of September
depending on the sowing date) were sterile with very small white
anthers.
 The mean minimum temperature during the period was 240C with
continuous and good sunshine hours. As the temperature reduced
towards the end of September and beginning of October, the flowers
started turning fertile with yellow anthers.
 The flowers turned completely fertile with almost full yellow
anthers when the mean minimum temperature reduced to 180C
during the following month.
 Highest fertility was recorded during the months of October-
November which later decreased as the temperature went down
further.

Fig.4: Sterile and fertile flower in TGMS tetraploid
29
Sterile flower Fertile flower

Week/ Month Flower behaviour
Weekly mean mini
temp (0C) Weakly mean sunshine hours
August, 2008
1st week Sterile 24.3 0.64
2nd week Sterile 24.7 Sunshine was nil during 4 days
3rd week Sterile 24.5 4.2
4th week Sterile 24.0 7.0
September 2008
1st week Partially fertile 23.8 4.7
2nd week Partially fertile 23.6 3.7
3rd week Partially fertile 24.2 5.0
4th week Partially fertile 22.5 7.5
October, 2008
1st week Partially Fertile (40% ) 23.3 7
2nd week Fertile (70-80 % ) 21.8 7.6
3rd week Completely fertile 18 9
4th week Fertile (80%) 15 9
November, 2008
1st week Fertile (80%) 14.0 9.0
2nd week Fertile (80%) 15.5 8.5
3rd week Completely Fertile 20.3 6.5
4th week Fertile (60-70%) 12.4 9.8
Table 3. The duration of sunshine hours and sterility/fertility observed in TGMS
line

Week/ Month Flower behaviour
Weekly mean mini
temp (0C) Weakly mean sunshine hours
August, 2009
4th week Partially fertile 23.90 4.6
September 2009
1st week Partially fertile 24.50 4.5
4th week Sterile 25.00 8.7
October, 2009
4th week Completely Fertile 16.0 10.4
November, 2009
1st week Completely Fertile 16.5 10.0
2nd week Completely Fertile 21.0 3.8
3rd week Completely Fertile 17.7 5.0
4th week Fertile 12.5 8.5
Conti…..

C. Transgenic genetic male sterility
 The male sterility induced by the technique of genetic engineering is
called transgenic male sterility.
 The gene ‘barnase’causing male sterility is integrated into 'A' line.
 Another gene 'barstar' suppresses the activity of male sterility gene
barnase and hence can be used for fertility restoration.
 The transformation frequency of 1-2% has been observed. The
transgenic plants upto 50% have been reported to be stable in
phenotype (male sterility) and in certain cases up to 90% of the
transgenic plants were male sterile and were stable.
 Hence, this technique can be utilized for commercial hybrid seed
production in near future in cotton.

32
C. Transgenic genetic male sterility
33
Source: https://www.slideshare.net/transgenic-male-sterlity

Utilization of GMS in plant breeding
1. Large number of parents can be used in crosses, because
the genotypes have dominant and recessive genes for male
sterility.
2. GMS generally does not have undesirable agronomic
characters.
3. It is possible to breed the varieties from segregating
population of GMS.

How to Differentiate Male Sterile &
Male Fertile Plant ?
34

2. Cytoplasmic male sterility (CMS)
 Governed by cytoplasmic genes
 Cytoplasmic male sterility was first
reported by WELCH and
GRIMBALL, 1947
 Includes ‘A’ (MS) and ‘B’
(Maintainer) line system.
 Result of mutation in
mitochondrial genome (mtDNA).
 Utilization of CMS in Plant
Breeding
This type of male sterility is of
importance in certain ornamental species
where the vegetative part is of economic
value.
37
India Singh (2015)

3. Cytoplasmic genetic male sterility (CGMS)
 Cytoplasmic genetic male sterility
was first reported by JONES and
DAVIS in onion
 Pollen sterility which controlled by
cytoplasmic and nuclear genes
 It consists of A, B and R lines
 Male sterility is maintained by
crossing with maintainer line
 Also known is Nucleoplasmatic Male
Sterility
Male Fertility
38
India Singh (2015)

A - Line B - Line
R - LineA - Line
Hybrid Seed
(Male Sterile)
(Male Sterile)
(Male Fertile)
(Male Fertile)
F1
Fig.4 : Mechanism of fertility restoration in CGMS hybrid lines
♀ ♂
♀ ♂
39

1. In CGMS system, CMS is highly stable and is not affected by
environmental factors.
2. In CGMS system, CMS 'A' line gives only male sterile plants.
3. CMS requires less area for maintenance.
4. The quantity of seed produced is more.
5. There is no chance of admixture.
Utilization of CGMS in plant breeding
40

Accessibility in Genetic Resources
41

Table 4.Male sterility genes identified in cotton
Gene Species Identified By
ms1 G. hirsutum Justus and Leinweber, 1960
ms2 G. hirsutum Richmond and Kohel, 1961
ms3 G. hirsutum Justus et al., 1963
MS4 G. hirsutum Allison and Fisher, 1964
ms5ms6 G. hirsutum Weaver, 1968
MS7 G. hirsutum Weaver and Ashley, 1971
ms8ms9 G. hirsutum Rhyne, 1971
MS10 G. hirsutum Bowman and Weaver, 1979
ms14 (Dong A) G. hirsutum Tianzhen et al., 1994
ms15 (Lang A) G. hirsutum Tianzhen et al., 1994
ms16 (81 A) G. hirsutum Tianzhen et al., 1994
MS11 G. barbadense Turcotte and Feaster, 1979
MS12 G. barbadense Turcotte and Feaster, 1985
ms13 G. barbadense Percy and Turcotte, 1991
ams1 G. arboreum Singh and Kumar, 1993
ar.ms G. arboreum Meshram et al., 1997 42
Bulletin
no: 24 Anon., 2002

Sources of male sterility
These are important sources of male sterility viz.,
• Interspecific crosses
• Spontaneous mutations
• Induced mutations.

Interspecific crosses
 The cytoplasms of three diploid species, viz.,
G. anomalum, G. harknessii and G.arboreum interact
with nuclear genes of G. hirsutum and produce male
sterility.
 The cytoplasm of G. arboreum and G. anomalum are
heat-sensitive and therefore less stable.
 The cytoplasm of G. harknessii and genome of
G.hirsutum interaction produce stable and dependable
cytoplasmic male-sterility in cotton over all
environments.

Sources of male sterility in cotton
Source of ms cytoplasm Nuclear genome
G.anomalum,G.arboreum,G.harknessii G.hirsutum
G.anomalum,G.arboreum Heat sensitive,less stable
G.harknessii × G.hirsutum Stable cms all over the environment
New sources of CMS Genome
G.harknessii × G.hirsutum CMS 2 (D2)
G.aridum Skovt. × G.hirsutum CMS 4 (D4) (PDKV, Akola)
G.trilobum × G.hirsutum CMS 8 (D8) (USA)
G.sturtianum × G.hirsutum CMS-C1
New sources of CGMS Genome
G.anomalum × G.thurberi Cg-mst (PDKV, Akola)

GMS lines in cotton
46
G-67 GMS,
Laxmi GMS,
IAN579 GMS,
B59-1684 GMS,
B-1007 GMS,
MCU-4 GMS,
MCU-5 GMS,
K34007 GMS and
Gregg GMS (MS 399)
DS 5 GMS (Hissar)
GAK-423A (Akola) etc.

Hybrid developed based on GMS system
47
Suguna : Worlds 1st MS based hybrid, CICR, RS Coimbtore, Gregg 399
x K3400 (Russian Var.), Medium staple (25mm) early maturity
(145 days)
AAH-1 : Intra arboreum, early maturing, high yielding, superior fibre
qualities, low seed setting (25%), North zone
• New hybrids
Moti (PAU)
Raj DH 9 (Rajasthan)

Improved CMS lines developed in cotton
48
JCMSBN
JCMSK2
S27CMS
DESHAMS-16
DESHAMS-277 and
Laxmi CMS
CAK 32
601CMS etc.

Sources of restorers identified
49
MEX 685-3
Dixieking ne restorer
Demeter III (I)
Demeter 2
DES 146C etc.

Hybrids developed based on CMS
50
MECH 4 :World’s 1st cotton hybrid, Mahyco, (601CMS x C219)
Maharastra, 1990
PKV Hy 3 : 1st hybrid released by public sector, intra hirsutum hybrid,
(CAK 32 x DHY 286-1R), 10-15% high yield over
conventional, resistant to jassids, Vidarbha region
PKV Hy 4 : 1996, (CAK 32 x AKHO 7R), extra long staple (30mm),
average yield 20 q/ha under rainfed, resistant to jassids
and mature 15 days earlier (165 days) than PKV Hy 3,
Vidarbha region

Mutations
 Mutations are of two types viz., spontaneous and induced.
 Spontaneous male sterility has been observed in upland and
arboreum cottons.
 In G. arboreum, the first spontaneous male sterility mutant
was identified in variety DS-5 at Haryana Agricultural
University, Hisar . The gene is designated as ams1.
 The semi and complete male sterility has been isolated by
various workers from the material treated by X-rays, gamma
rays and Ethyl Methane Sulphonate (EMS).
 At Dharwad, cotton variety Abadhita was treated with
gamma rays and various concentration of EMS.

Conti…..
52
Abadhita
Treated with double mutagen
at 10 kR gamma rays + 0.2 per
cent EMS
M1 generation male sterility
In M2 generation, the
segregation pattern of male
sterile to fertile was 1:1
Result indicate that presence
of GMS which was
conditioned by post meiotic
pollen abnormality.Bulletin no: 24 Anon., 2002

Induction of male sterility by chemical
 Male sterility can be induced through the use of chemicals,
which are commonly known as male gametocides.
 Some of the chemicals used for induction of male sterility is
FW 450 (Sodium-dichloro-iso-butyrate) or MH-30 (Maleic
hydrazide) and Ethidium bromide (a potent mutagen).
 Spraying of aqueous solution of FW-450 or MH30 induces
male sterility in cotton.
 Higher concentration of treatments caused male as well as
female sterility and various adverse effects like reduction in
yield, boll and seed size and increase in lint percentage.

Conti..
Chemical Concentration
(%)
Pollen
sterility (%)
FW-450 (Sodium-dichloro-iso-
butyrate)
1.5 76.3-97.8
MH-30 (Maleic hydrazide) 1.0 70.1-80.2
Ethidium bromide 3.0 67.6-78.0
*All chemicals applied during bud initiation or at anthesis
54India Singh et al. (1989)

Conversion into male sterile lines
 The potential female parents of hybrids can be converted into
male sterile lines. To convert in GMS background, four to five
backcrosses accompanied by alternate selfing is required.
 Finally, a line which is heterozygous for sterility and showing
1: 1 segregation is selected having all the characters of the
recurrent parent.
 The local genotypes with good agronomic backgrounds can be
converted into CMS line by attempting five to six backcrosses.
 Many promising varieties and elite germplasm lines have been
converted into CMS and restorer lines and few in GMS
background

Table 5.Genotypes converted into GMS and CMS
background
Male Sterility
System
Genotypes
GMS (4x) LRA 5166, SRT 1, DGMS 1, HGMS 2, GAK 32A, SHGMS-9,
DGMS2,SHGMS-5
CMS Germplasm - G 67, DMS A-8, RCMS A-2. GSCMS-15, 34, DMSA
15,CAK32A,C1412,C 1998, CAK 1234, LCMS 6, JK 119,,IC 1547
Varieties - Rajat, LH 900, Supriya , G. Cot l0, Laxmi , Abadhita, BN,
K2,LRA 5166, H 777, G. Cot 14, Ganganagar Ageti, F 414, Bhagya,
Kh3, Narmada,Deviraj
GMS (2x) GMS 4, GMS 2, GAK 20A, GAK 09, SGMS 2, SGMS 4, RGMS A-
2,RGMS 3,SGMS 13, GMS 4-1, GAK 15A, GAK 26A, Sujay, GAK
423,GAK 8615
R line NH 258, AKH 545, GSR 22, AKH 39R, LR 29, AKH 26R, AKH
1167,GSR 6,DR 6, DR 1, AKH-01-143, LR 104

Role of honey bees in hybrid seed production
through male sterility
 Cross-pollination in cotton takes place through insects. The honey
bees (Apis mellifera) are major pollinators. These pollinating
agents when used in hybrid seed production of hybrids based on
male sterility may help in significantly reducing the cost of hybrid
seed.
 On an average 10 bees are reported to be sufficient to pollinate 100
cotton flowers. The yield of seed cotton in ‘A’ line depends on the
bee population and distribution of bee colonies.
 It was found that 13- 15 bee colonies are adequate for economical
seed production. However, studies conducted in the central zone
indicated that hand pollination was better than insect or bee
pollination for boll setting and seed quality.
57
Mofett et al. (1976)U.S.A.

Conti…..
 The reasons attributed to this are shift in visit of honey bees from cotton
to other more rewarding crops in the area, higher gossypol content in
cotton and heavy use of pesticides.
 Few basic studies show that sucrose content in the flower nectar
especially in CMS flowers affected significantly the honey bee population
density and might be a key factor for higher hybrid seed production in
CMS lines.
 Among various sugar components of the cotton nectar, only sucrose
showed highly positive and significant correlation with frequency of
visits by honey bees.
 Thus breeding lines with higher sucrose content is needed for increasing
bee visits which in turn can increase the boll setting per plant and
decrease the number of aborted seed, resulting in higher yield of hybrid
seed.
58

Cost of Hybrid Seed Production
59

Table 6.Cost of seed production of GMS based intra-hirsutum
hybrid (Study conducted at HAU, Hisar )
Sr.
No Particulars / Items
Actual cost incurred (Rs.)
GMS based
hybrids
Hand
emasculated
hybrids
1 Cost of cultivation 318 318
2 Parental seed cost 79 79
3 Emasculation / Pollination charges 560 1120
4 Delinting grading and packing charges 60 60
Total cost 1017 1577
Hybrid seed obtained (kg.) 10.00 2.880
Net cost per kg Rs. 102.00 Rs.547.00

Table 7. Cost of seed production of CGMS based intra-
hirsutum hybrid (Study conducted at PDKV, Akola)
Sr.
No Particulars
Cost (Rs.) Average of 3 years
Hand emasculated
hybrids
Male sterility
1 Total cost of seed production
(Labour, Materials,
cultivation, insecticide,
fertilizer )
42566.27 16973.20
2 Net cost per kg Rs.192.59 Rs.31.51
3 Yield of seed cotton (kg/ha) 350.13 678.22

Case Study
on
Genetic Male Sterility
62

Table 8. Mean performance of top two hybrids and checks for
different characters in cotton of rainfed condition
Characters Crosses Checks
Million-GMS
× Jayadhar
GAKA -423
× H-221
Ch-222 DLSa -17 Rahs- 14
Plant height (cm) 163.05 157.20 149.65 133.60 128.60
Number of monopodia per Plant 27.50 28.00 26.20 25.60 28.40
Number of sympodia per Plant 2.70 3.30 2.80 3.10 3.80
Number of bolls per plant 20.85 28.25 13.30 20.45 5.85
Boll weight (g) 2.40 2.20 3.70 2.25 1.76
Seed cotton yield per plant (g) 49.95 60.25 49.65 44.70 7.80
Ginning outturn (%) 30.92 33.71 40.65 44.70 7.80
Lint index (g) 2.68 2.53 4.90 3.18 3.32
Seed index (g) 5.95 5.89 7.03 6.06 7.02
2.5% span length (mm) 26.10 25.30 22.90 25.20 22.90
Fibre strength (g/tex ) 22.80 20.90 15.50 20.20 17.60
Seed cotton yield (kg/ha) 929.00 780.61 652.80 513.37 290.08
Dharwad Jyotiba et al. (2010) 63

Table 9. Estimation of standard heterosis for seed cotton yield
and its component characters in cotton
Hybrids/Cross Seed cotton
yield
Bolls/Plant Monopodi
/Plant
Seed index GOT 2.5 per cent span
length
GMS 4 x LH 2076 32.1* 23.4* 43.3 15.0* 3.1* 2.6
GMS 4 x F 1861 30.4* -1.2 33.3 20.3* -1.4 -1.8
GMS 4 x 002 NAH 5.4 35.8* 10.0 7.8* 2.5 -0.4
GMS 20 x RS 2013 -3.6 30.5* 256.7* 0.0 -2.1 -1.
GMS 4 x 0238 DA -7.1 33.6* 23.3 17.6* -1.4 2.6
GMS 26 x 002 NAH -23.2 30.5* 123.3 -11.1 9.0* 1.8
GMS 4 x RS 810 -30.4 31.3* 46.7 3.9 -2.1 -2.6
GMS 20 x F 1861 -37.5 21.3* 266.7* 9.8* -1.4 0.0
GMS 16 x Biyani 161 -41.1 -2.1 266.7* 14.4* -0.8 5.4*
GMS 20 x 002 NAH -75.7 -7.4 66.7 11.1* -6.0 6.7*
GMS 27 x 002 NAH -82.1 18.1* 110.0 28.1* -3.4 2.6
GMS 20 x LH 2076 -83.9 -37.4 0.0 23.5* 3.8* -3.6
Sirsa Tuteja et al. (2011)Zonal check = CSHH 198, GOT = Ginning outturn (%) ; 64

Table 10. Per cent heterosis for fibre quality traits over mid
parent (MPH), better parent (BPH) and check
(Jayadhar)
Sr.
No.
Crosses 2.5 % span length Fibre strength
MPH BPH Jayadhar MPH BPH Jayadhar
1. MSD 7 nor x ARBH-35 (a x a) 2.64 0.40 2.85 19.83** 10.20** 11.92**
2. MSD 7 nor x DDhc 11 (a x h) 5.25* 1.87 -0.20 37.20** 36.78** 16.58**
3. MSD 7 nor x Jayadhar (a x h) 3.08 2.03 2.03 0.70 -6.74* -6.74*
4. MSD 7 nor x RAhS-14 (a x h) 18.94** 16.60** 14.23** 16.37** 12.11** 3.11
5. MSD 7 nkd x DLSa-102 (a x a) -8.82** -8.82** -5.49* -3.49 -4.01 -6.99**
6. MSD 7 nkd x Jayadhar (a x h) -13.97** -15.49** -12.40** -21.84** -23.06** -23.06**
7. MSD 10 nor x RAhS-14 ( a x h) 7.96** 2.32 7.52** 9.97** 2.95 8.55**
8. MSD 11 x DLSa-102 ( a x a) -13.13** -13.73** -10.57** -3.99 -5.68* -9.59**
9. MSD 11 x ARBH-35 ( a x a) 4.87* 4.76 7.32** -3.07 -7.40** -5.96*
S.Ed. 0.56 0.64 0.64 0.42 0.49 0.49
C.D. (P=0.05) 1.12 1.30 1.30 0.86 0.99 0.99
C.D. (P=0.01) 1.50 1.73 1.73 1.14 1.32 1.32
Dharwad Geddam and Khadi (2013)
* and ** indicate significance of values at P=0.05 and P=0.01. respectively.
65

Table 11. Mean performance and estimates of economic heterosis in
crosses of desi cotton for seed cotton yield and component
traits
Cross Seed cotton yield Bolls/ plant (g) Seed index Ginning outturn (%)
Mean Heterosis Mean Heterosis Mean Heterosis Mean Heterosis
DS 5 x HD123 2672* 10.10 64.70 9.90 5.40 4.40 40.39 -0.60
DS 5 x 1-24 3433* 41.40 75.55* 28.30 4.80 -7.10 41.33 1.60
DS 5 x AC 3079 3840* 58.20 72.30* 22.80 4.97 -3.80 39.56 -2.70
DS 5 x LD 153 2889* 19.00 66.40 12.80 5.33 3.06 43.85* 7.80
DS 5 x 30861 BLL 3047* 25.40 71.85* 22.00 4.75 -8.10 40.97 0.70
DS 5 x Sanguniun 2234 -8.10 57.00 -3.10 5.45 5.40 38.59 -5.10
DS 5 x N 67/92 2470 1.70 61.75 4.90 5.55* 7.30 37.12 -8.70
DS 5 x PR18/91 2852* 17.50 68.10 15.70 5.42 4.80 37.24 -8.40
DS 5 x B31/92 2430 0.00 55.80 -5.10 5.65* 9.20 36.89 -9.20
DS 5 x AKA8401 2312 -4.80 49.60 -15.70 5.25 1.50 37.39 -8.00
DS 5 (C) 2430 58.85 5.17 40.67
C. D. (P=0.05) 149.81 12.90 0.32 1.03
* Significant at P=0.05Hisar (Haryana) Singh et al. (2013) 66

Table 12. per se performance of hybrids for seed cotton yield
and its component traits in cotton
Sr.
no
Parents/crosses Seed cotton
yield (kg/ha)
No of bolls
per plant
Ginning
out turn
(%)
2.5% span
length (mm)
1 GMS 20 X SA 1652 1763 41.3 35.4 26.6
2 GMS 17 X MC 127 1770 47.7 35.0 25.8
3 GMS 26 X MC 88 1773 47.0 32.5 27.7
4 GMS 26 X CSH 3129 1730 55.7 33.8 27.6
5 GMS 17 × SA 1652 710 39 36.6 23.5
6 GMS 20 × MC 88 1944* 43.7 35 25.7
7 GMS 20 × CSH 3129 802 57 34.6 27
CSHH 198 (C) 1698 36.3 33.1 26.9
CD (%) 177 9.85 1.00 2.31
CV (%) 10.16 14.50 1.75 5.24
Sirsa (Haryana) Tuteja and Agrawal (2013) 67

Table 13. The estimates of heterosis over standard check ( SC ) under pooled
over environments (Surat, Bharuch and Hansot) for different
characters in Asiatic cottons.
Sr.
no
Crosses Number of
monopodia
per plant
Number of
open bolls
per plant
Ginning
percentage
( % )
Seed cotton
yield per
plant
1 378 BK X 824 25.91** 28.04** -2.81 29.89**
2 4011 X 824 23.18** 21.15** -1.02 26.21**
3 35N X G.Cot.17 22.58** 23.66** -2.30 25.77**
4 35N X 824 17.58** 22.68** 0.92 18.14**
5 4011 X G.Cot.17 19.55** 18.68** -4.06* 17.63**
6 4011 X GShv 1012/90 22.88** 22.64** -6.76** 16.41**
7 8401 X G. Cot. 17 21.21** 17.94** 0.00 13.65*
S.Ed. 0.22 1.47 0.55 5.06
CD @ 5 % 0.53 3.47 1.29 11.97
CD @ 1 % 0.78 5.14 1.91 17.72
Navsari (Gujarat) Patel (2014)
Standard check : G.Cot MDH 11, * - Significant at 5 % level , ** - Significant at 1 % level
68

Table 14. Per cent heterosis over better parent (H1) and standard check (H2)
for various traits
sr.
no
Crosses No. of bolls/ plant 2.5 % Span
length
Fibre fineness Seed cotton
yield / Plant
H1 H2 H1 H2 H1 H2 H1 H2
1. GAK-8615 x
AKA-7
37.70* 21.15 -5.31* 4.35 2.84 0 51.28** 28.26*
2. GAK-8615 x
AKA-5
53.83** 35.34** -5.93* 3.67 19.12** 11.72 45.13** 18.84
3. GAK-8615 x
AKA-0209
5.07 22.6 -8.77** 0.54 -2.84 -5.52 44.74** 19.57
4. GAK-8615 x
AKA-9703
63.39** 43.75** 2.96 13.47** 7.8 4.83 -23.36 -23.91*
5. GAK-8615x
AKA-9009-1
24.04* 32.69* -4.69* 5.03* 0.71 -2.07 17.46 7.25
6. GAK-423 x
AKA-5
29.46 4.57 1.93 7.62** 3.01 -5.52 25 15.94
S.Ed. 3.47 3.47 0.60 0.60 0.29 0.29 5.33 5.33
CD at 5% 7.08 7.08 1.22 1.22 0.60 0.60 10.89 10.89
CD at 1% 9.54 9.54 1.65 1.65 0.81 0.81 14.67 14.67
Akola (Maharashtra ) Solanke et al. (2015) 69Check = PKV Suvarna

70
Sr.
no.
Top five
hybrids
Heterosis value
Seed cotton
yield/plant
No. of boll Boll weight/boll
C.C. M.P. B.P. C.C. M.P. B.P. C.C. M.P. B.P.
1. DGMS 34 x
HD 517
(125.68)
53.19 158.35 82.52 41.05 144.71 73.11 11.17 6.79 3.03
2. DGMS 34 x
HD 523
(114.86)
40.00 84.85 66.8 20.83 65.54 48.30 13.00 14.29 12.66
3. DGMS 2 x
HD 528
(144.83)
39.96 128.68 121.8 24.85 114.31 111.23 13.65 19.63 18.54
4. DGMS 1 x
HD 432
(114.34)
39.37 124.75 114.9 27.16 136.10 132.77 10.00 17.59 16.45
5. DGMS 1 x
LD 1026
(103.03)
25.58 82.92 60.76 15.92 78.85 51.54 12.25 13.67 14.96
Table 15. Top five hybrids based on mean of yield traits and their heterosis
values
Vekariya et al. (2017) 75Hisar (Haryana) Check = AAH 1

Case Study
on
Cytoplasmic Genetic Male Sterility
71

Table 16. Per cent heterosis of selected hybrids over check for
different characters
Hybrid Bolls/plant Boll weight Seed cotton yield
MS 1 x TR 34 18.9 2.7 73.0**
MS 2 x TR 10 -23.2 25.8* 48.1
MS 1 x TR 9 -25.2 33.5** 51.2
MS 2 x TR 101 -7.1 8.8 54.7*
MS 3 x TR 14 -48.0** 10.4 -10.6
MS 3 x TR 23 -28.8 37.3** 39.9
MS 3 x TR 101 7.9 14.2 90.0**
MS 4 x TR 14 39.2* 54.0 95.1**
MS 5 x TR 4 -20.8 31.9** 55.9*
MS 5 x TR 14 32.3 -5.0 86.4**
MS 5 x TR 16 8.7 5.0 73.9**
MS 5 x TR 23 -20.7 39.6** 66.6*
MS 5 x TR 31 -35.4* 11.5 9.9
MS 6 x TR 14 18.1 -16.5 62.0*
SE 2.83 0.27 7.32
Nagpur Gill et al. (2009)
72
Check = F 1861

Table 17. per se performance of hybrids for yield and other characters
HYBRID 2.5% Span
Length(mm)
S.I. (g) GOT (%) Seed cotton yield (kg/ha)
Anjali x SR 38.1 13.9 28.4 1709
LRA 5166 X SR 37.2 12.4 28.3 1549
70 G X SR 32.5 10.4 32.6 1229
IRH 1-10 X SR 31.8 10.2 30.6 1044
IRH 1-6 X SR 31.2 9.4 33.4 970
Laxmi X SR 32.5 9.2 31.7 866
IRH 1-4 X SR 31.1 8.6 34.5 776
MCU 5-VT X SR 31.9 9.6 31.5 727
Abadhita X SR 31.9 8.5 32.3 681
Suman X SR 30.1 9.1 29.7 553
70 E X SR 31.7 9.1 31.3 532
70 E X PR 35.6 9.9 34.4 512
DCH 32 [C] 36.8 12.9 31.9 1197
Sruthi [C] 32.7 9.5 31.5 884
CD @ 5% 0.3 0.6 0.9 454
CV % 8.2 10.3 12.0 13.1
Coimbatore
Where S.I.= Seed index, SR = Suvin Restorer, GOT = Ginning outturn (%) and C= Check ;
Manickam et al. (2010) 73

74
Table 18. Per cent heterosis of top hybrids for different characters in cotton
Sr
.n
o
Pedigree Day to flowering No.of bolls/plant Ginning per cent Seed cotton
yield/plot
Mean SH Mean SH Mean SH Mean SH
1 1A × 054 52.5 -5.41* 38.0 5.56 37.70 6.35** 0.69 18.97**
2 1A × 058 54.5 -1.80 41.0 13.89* 35.90 1.27 0.72 24.14**
3 10A × 054 54.5 -1.80 35.0 -2.78 37.65 6.21** 0.72 24.14**
4 9A × 054 54.5 -1.80 23.0 -36.11 37.95 7.05** 0.64 10.34*
5 9A × 056 56.0 0.90 36.30 0.00 37.95 4.23* 0.67 15.52**
6 3A × 054 55.5 0.00 38.0 5.56 37.20 4.94* 0.72 24.14**
7 8A × 056 58.0 4.50* 37.0 2.75 37.40 5.50** 0.70 20.69**
8 8A × 058 59.0 6.31** 21.0 -41.67* 37.90 6.91** 0.65 12.07*
PKV Hy-
4©
55.5 36.0 36.48 0.58
G.M 56.4 32.38 36.83 0.57
SE.M± 1.25 2.18 0.65 0.04
CD at 5% 2.51 4.40 1.32 0.08
CD at 1% 3.42 5.90 1.78 0.11
Rahuri (MH) Shinde et al. (2012)
74

Table 19. Estimation of fertility restoration in CGMS based intraspecific
hybrid of G.hirsutum
CMS lines Restorer lines
RHCr-035 RHCr-054 RHCr-056 RHCr-058
RHCCMS-1A 96.89 98.19 94.77 97.90
RHCCMS-2A 95.12 96.43 93.92 93.01
RHCCMS-8A 90.71 94.48 92.00 95.37
RHCCMS-9A 95.56 96.68 93.49 97.90
RHCCMS-10A 90.72 95.67 92.81 93..10
RHCCMS-3A 87.16 94.78 93.24 89.52
RHCCMS-4A 93.10 94.36 90.86 91.02
RHCCMS-6A 94.75 93.01 89.42 95.68
RHCCMS-7A 95.95 94.88 92.71 91.75
Mean 93.33 95.39 92.58 94.25
Rahuri (MH) Shinde et al. (2012)
75

Table 20. per se performance of CGMS based hybrids for
yield and its component traits in upland cotton
CMS based hybrid Seed cotton
yield (kg/ha)
No. of
monopods
No. of sympods Gining
outturn (%)
2.5% span
length (mm)
K34007 x CIR 70 2693* 10.0* 9.0 32.3 26.8
F 1183 x CIR 26 2474 9.7* 7.0 30.7 27.6
RB 281 x CIR 38 2440 8.1 8.1 35.3* 26.5
F 1183 x CIR 38 2070 2.9 14.0* 34.0 26.8
K 34007 x CIR 32 2019 7.7 8.3 34.4 29.0*
LRA 5166 x CIR 12 1682 7.7 9.1 33.0 29.1*
Jhorar x CIR 38 1464 6.0 8.6 33.7 29.1*
LH 1134 x CIR 23 1346 5.6 8.0 35.6* 29.2*
SH 2379 x CIR 23 1001 11.0** 9.0 35.9* 29.0*
F 1183 x CIR 23 891 5.8 9.6 35.7* 28.3*
CSHH 198 (Check) 2168 6.7 11.2 33.0 26.6
CD at 5 % 330.7 1.8 2.8 2.1 1.7
CV 11.92 18.6 3.9 3.9 3.8
Tuteja (2014)
* indicate significance of values at P=0.05. respectively.
Sirsa (Haryana) 76

Table 21. Main features of hybrids released using male sterile line
Name of hybrid Year of
release
Yield
(q/ha)
GOT
(%)
MFL
(mm)
Spinning
counts
Area in which
released
Type of
hybrid
SUGUNA 1978 30 35 25 40 TamilNadu HH
ANKUR 15 1983 30 35 26 50 Vidarbha HH
MECH 11 1984 25 38 28 50 MS and MP HH
MECH 4 1990 25-30 35 29 50 MS,GS,MP,RS HH
PKV HY. – 3
(CAHH 468)
1993 15R 36 25 40 Vidrabha HH
PKV HY. – 4
(CAHH 8)
1996 20R 35 30 50 Vidarbha HH
ANKUR 9 1997 30 37 27 40 MS,GS,MP HH
AAH 1 1999 24 38 16 <10 Haryana AA
PKV HY-5 2000 15R 35 26 40 Vidarbha HH
AKDH 7 2000 12R 38 24 30 Vidarbha HH
G.COT MDH 11
(GSGDH 2)
2002 20 36 23 27 Gujarat AA
Achievements
77

Conclusion
 It is concluded that hybrid seed is an important for
higher yield, better fiber quality and other related
attributes in cotton. To reduced cost of hybrid seed
production and thrust of quality seed, male sterility
based hybrid seed production is the best alternative.
It also helps to exploits the hybrid vigour to a greater
extent for genetic yield potential of cotton.
78

Problem associate with male sterility in cotton
 GMS is less stable sometimes, sterile plants become fertile under
low temperature conditions.
 In GMS the lines segregate into male sterile and fertile plants in 1:
1 ratio.
 Conversion of a genotype into GMS (ms5ms6) needs selfing after
each backcross to isolate recessive genes and hence more number
of generations are required.
 There is possibility of admixture if fertile plants are not properly
rogued out.
 In CGMS drawback of cytoplasmic effect and fertility restoration.
79
Source: http://krishikosh.egranth.ac.in/Gaddem/Thesis
CICR Technical Bulletin no: 24

Male sterility in cotton

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