Development of male sterile lines in brinjal and chilli.pptx

1. Development of Male Sterile
Lines in Brinjal and Chilli

2. Male Sterility
• It is the failure of plants to produce
functional anthers, pollen, or male gametes.
Manifestations
– Chromosomal aberrations
– Gene action or cytoplasmic influences
– Failure to develop normal microsporogenesis tissue-
anther
– In modification of entire flower, stamens or pistils
– Abnormal pollen maturation
– Role of esterase

3. Classification
• Genetic male sterility e.g. chilli, muskmelon
• Cytoplasmic male sterility e.g. onion, cole crops
and carrot
• Cytoplasmic genic male sterility e.g. brinjal and
chilli
• Functional male sterility e. g. tomato and brinjal
• Gametocidal male sterility
• Genetically engineered male sterility e. g.
Brassica sps

4. HISTORY OF Male Sterility
GMS has been reported in cabbage by Rundfeldt, cauliflower by
Nieuwhof (1961)
Male sterility systems have been also developed through genetic
engineering (Williams et al. 1997) and protoplast fusion ( Pelletier
et al. 1995)
1960
Pelletier
1995
Rundfeldt

5. Genetic Male Sterility
• Reported in about 175 plant species (Kaul 1988)
• Usually recessive and monogenic
• No undesirable agronomic characters
• Temperature sensitive
• 50% of fertile plants to be removed from field
• Availability of marker gene- closely linked with
ms gene

6. Maintenance of GMS
Line A
Male sterile
msms
Line B
Maintainer Line
Msms
(X) Msms : msms
Male fertlile Male sterile
Msms
(X) Msms : msms
Male fertlile Male sterile
Msms
And so on ……

7. Use of GMS for hybrid development
Crops Gene Commercially
utilized
Variety
Tomato Single recessive
gene
Ps-2 gene -
Chilli Single recessive
gene
Ms-12 & ms-3 gene CH-1, CH-3
Muskmelon Single recessive
gene
Ms-1 gene Punjab hybrid -1

8. Cytoplasmic male sterility (CMS) systems
 Determined by cytoplasm
 Result of mutation in the mitochondrial genome (mt-
DNA)
 Easily transferable trait
 Highly stable
 Not use where seed is the economic part
 Inferior agronomic performance

9. S F
rr
rr
S
rr
S
F
S
F
rr
rr rr
rr
Male sterile
Male sterile
Male sterile
Male fertile
Strain B
Strain B
Repeated Backcross
F1
BC1
BC 6or BC 7

10. Interaction of cytoplasm and nuclear genes
Genotypes of CGMS lines
Cytoplasmic genetic male sterility
S
F
S
F F
S
RR RR
rr
Rr
rr
Rr
RR gene with F cytoplasm
Fertile R- line
RR gene with S cytoplasm
Fertile R- line
Rr gene with F cytoplasm
Fertile
Rr gene with S cytoplasm
Fertile
rr gene with F cytoplasm
Fertile B- line
rr gene with S cytoplasm
Sterile A line

11. • S rr rr x N rr rr All sterile
• S rr rr x N Rr rr Fertile 50% ; Sterile 50%
• S rr rr x N/S Rr Rr All fertile
Crops Gene Commercially
utilized
Variety
Chilli Single recessive
gene
ms-2 CH-52, Arka
Meghana, Arka
Shweta, Arka Harita,
Kashi surkh
Onion Single recessive
gene
- Arka Kirtiman, Arka
lalima
Carrot Single recessive
gene
- Pusa Nanjyoti, Pusa
Vasuda

12.  By progeny performance or crossing with a few normal
genotype
 Some rows may consist of all fertile and some rows
sterile & fertile plants may occur in 1:1 ratio : GMS
 All the progenies in all the rows may be sterile :
CMS
 Some rows may have all fertile plants, some all
sterile plants & some have fertile and sterile plants
in 1:1 ratio : CGMS
Detection of MS system

13. Steps in development of male sterile line
1. Morphological and molecular evaluation of male sterility system.
2. Studies on blossom biology on floral morphology of male sterile and fertile flowers.
3. Transfer of male sterility into different genetic backgrounds to identify corresponding maintainer &
restorer lines/ pollen parents.
4.Studies on microsporogenesis and megasporogenesis of male sterile and male fertile lines.
5.Development mapping population (parents, F1, F2 ,BC1, BC2 progenies) for genetic studies.
6. Identification of molecular markers
7. Studies on reproductive biology and identify causes of male sterility (CMS,CGMS& GMS system)).
8. Use of diagnostic PCR kit to identify sterile, maintainer and restorer lines.
9. Development of stable male sterile lines, maintainers, restorer and male parental lines to develop
uniform efficient and durable F1hybrids.

14. Achievements
GMS based hybrids:
CH-1 (MS 12 x Ludhiana long sel.)
CH-3 (MS 12 x S-2530)
Tomato Chilli Onion
Ludhiana Anand Banglore
ms33IPA CCA 4261 Ms 1
ms2 IPA CCA 4759 Ms2
ps2 L3841 CCA 4758 ms 3
ps2 San Pedro
ps2 UC 82-B Ludhiana
ms-45 CCA 4261
Hybrids Identified through AICRP- VC
CCH-2 (A1 x Pusa Jwala) and
CCH-3 (KA-2 x RPBC-473)
ARCH-228, Meghna
Male sterile genotype
available in India

15. Case studies

16. Rapid Methods of Improvement
in Brinjal
 Involvement of ORF from mitochondrial genes for
occurrence of CMS system (Hanson and Bentolila 2004).
 Variations in atp and cox genes in mitochondria of wild
Solanum species caused the male sterility (Yoshimi et al
2013).
 Rf genes for restoring of fertility of the plant (Bentolila
et al 2002)
 RAPD marker (OPAB10) was further converted into SCAR
marker (SCAB101900) and has potential application in the
selection of fertility restorer plants
Dhatt and Sidhu 2020

17. Development of Male-sterile lines in brinjal
Interspecific hybridization
S. gilo S.
kurzii S
violaceum
S.
virginianum
S.
aethiopicum
S. anguivi S.
grandifolium
Anther indehiscent type
S violaceum
S.
virginianum
S. Kurzii

18. Scheme for development of male-sterile lines (ms)
in brinjal

19. Scheme for development of fertility restorer (Rf)
lines in brinjal

20. Development of cytoplasmic male sterile lines in chilli
(Capsicum annuum L.) and their evaluation across multiple
environments
 A breeding program was initiated in 2009 to develop temperature stable CMS
lines in chilli.
 ‘CCA 4261’ was used as a CMS donor.
 Maintainer plants were identified from ‘SL 461’, ‘SL 462’ and ‘SD 463’.
 17 CMS lines in diverse genetic backgrounds were established.
 The CMS lines were evaluated for stability of sterility over four environments
during 2014–15 and 2015–16
Jindal et al 2018

21. Maximum and minimum weekly temperature recorded during period of pollen
viability study; A. during low temperature of 2014–15 (E1); B. during high
temperature of 2014–15 (E2); C. during low temperature of 2015–16 (E3); and
D. during high temperature of 2015–16 (E4).

22. Sterile and fertile phenotypes in F1 generation of 11
testcrosses
Testcrosses
Number of individuals/plants
Total Sterile Fertile % Male sterile
plants
CC 141 × MS 341 60 7 53 11.67
CC 141 × SL 461 60 43 17 71.67
CC 141 × SL 462 60 47 13 78.33
CC 141 × DL 161 60 32 28 53.33
CC 141 × EL 181 60 25 35 41.67
CC 141 × US 501 60 0 60 0.00
CC 141 × PA 401 60 0 60 0.00
CC 141 × SD 463 60 44 16 73.33
CC 141 × PP 402 60 0 60 0.00
CC 141 × PS 403 60 0 60 0.00
CC 141 × VR 521 60 4 56 6.67

23. Pedigree of CMS chilli pepper lines developed
in diverse genetic backgrounds

24. Male fertile flower of CMS B-
line ‘CMS463D14B’ showing
abundant of pollen grains
adhering to the green anthers
Male sterile flower of
CMS A-line ‘CMS463D14A’
showing purple anthers
without pollen grains;

25. Seed
Setting
ability was
normal
Wider choice of female
parent
Ten lines
CMS4611A CMS4622A CMS4624A CMS4626A CMS46213A
CMS463D2A CMS463D13A CMS463D14A CMS463L5A CMS4614A
Broaden
the CMS
germplasm
resources
in chilli.
Tolerance to temperature
variations
No association with flower
and fruit deformity
Results

26. Future Prospects
 Identification of new ms line
 Identification of potential restorers
 Pollination mechanisms of male sterility in different
vegetable crops should be further investigated.
 Multiple resistance/ tolerance to biotic as well as abiotic
stresses by transfer of genes using conventional and
biotechnological approaches.
 To identify potential markers for genetic purity testing.
 Potentiality of transgenic male sterility