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Male sterility in Cross Pollinated and
Vegetative crops
A.Manivannan
Scientist (Genetics)
DMR, New Delhi
Male sterility in Sunflower (Helianthus spp)
Genetic Male sterility (GMS)
Complete male sterility
ms1-ms5 = male sterility in sunflower recessive gene
Two types of g-mst
Type 1-gmst-Bloomington type
Type 2-gmst-Modern type
Cultivated Sunflower variety Karlik-68(Dwarf 68)- two recessive genes
msi1,msi2(Stable and complete male sterile)
Partial male sterility –p mst
Source of cms through Interspecific
Hybridization
x H.annuusN1
C1
F1 interspecific cross
H.grossessratus N1
C1
N1
C1
CMS- reccessive ms line
X
CGMS
H.petiolaris × H.annuus Repeated backcross of H.annuus
results in cms1 which is extensively
used mst in hybrid seed production
of sunflower all over the world
H.giganteus× H.annuus Cms3( S cytoplasm source)
H.annuus subspp
lenticularis × H.annuus
CV commander
Indiana 1
Chemical based
male sterility
Ethrel
GA
Mendok
TIBA
COTTON
Genetic Male Sterility (GMS):
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. However, male sterility governed by recessive genes is used in
practical plant breeding
All three types of male sterility occurs (g mst,c mst,gc mst) in cotton
sixteen different genes in tetraploid cottons (13 in G. hirsutum and 3 in G.
barbadense) and two in G. arboreum have been identified for genetic male
sterility.
Sterility is conditioned by dominant alleles at five loci viz, MS4, MS7, MS10,
MS11 and MS12 by recessive allele at other loci viz. msl, ms2, ms3, ms13, ms14
(Dong A), ms15 (Lang A) and ms16 (81 A).
Two male sterile phenotypes viz. ms5ms6 and ms8ms9 are conditioned by
duplicate recessive factors.
G. hirsutum line Gregg (MS 399) from USA is the basic source of
GMS possessing ms5 ms6 gene for male sterility.
GMS
CMS System
In case of CMS, the originally discovered CMS sources involving G.
arboreum and G. anomalum cytoplasmic systems having interaction
with ms3 locus were not found effective or stable under different
environments.
The only stable and dependable CMS source under varied environment
was developed through the utilization of G. harknessii. The complete
genome of G.hirsutum was transferred into the G. harknessii cytoplasm.
A single dominant gene ‘Rf’ from G.harknessii is essential for fertility
restoration.
Fertility enhancer factor 'E' for this CMS restorer system was obtained
from a G.barbadense stock.
The harknessii system is reported to contribute to good agronomic
properties and attraction to honey bees.
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
G. aridum Skovt. × G. hirsutum (D4)
G. trilobum × G. hirsutum CMS 8 (D-8)
G. sturtianum × G. hirsutum CMS-C1
New sources of CGMS
G. anomalum x G. thurberi Cg-mst
Mutation
G. arboreum, the first spontaneous male sterility mutant was identified in
variety DS-5
Chemical based male sterility
FW 450(Sodium B-Dichloro-iso-butyrate)
MH-30 (Maleic hydrazide)
Ethidium bromide
Male sterility based hybrid Production
GMS system. CPH2 (Suguna), First hybrid based on GMS released at
CICR, RS, Coimbatore
G. harknessii based cms with fertility restoration gene sources were used
in developing the hybrid CAHH 468 (PKV Hy-3).
T cytoplasm S cytoplasm C cytoplasm
Discovered by Rogers(1944) M.T. Jenkins Beckett(1971)
Developed
from
OP Mexican
maize variety
Gold june
Teopod maize Brazilian Maize
Male sterility Stable Unstable Stable
HMT
Susceptibility
Yes No No
Type of mst Sporophytic Gametophytic Sporophytic
Fertility
restoration
Fr1,Fr2 - Fr4,Fr5, Fr6
CMS
T cytoplasmic mitochondria, HM T pathotoxin causes the
uncoupling of oxidative phosphorylation, inhibiton of oxogluterate
oxidation and causes irreverisble swelling
Fertility restoration in maize
CGMS
Reported by Rhoades (1931) while working with
Peruvian source of maize cytoplasm
Interspecific crosses
Coix lacrymus-jobi X Z.mays
Euchlaena mexican X Z. mays
E.Perennis X Z.mays
Chemcial mst
GA, MH, Mendok, DPX 3778, Mo deficiency
A X B
(frfr) (FrFr)
ms mf
AB
(Frfr)
mf
Method of Hybrid seed production
Single cross Hybrid
T- cytoplasm required two Fr genes
C,S-cytoplasm requires one Fr genes
Most of the T cytoplasm posses one Fr gene
A X B
(frfr) (frfr)
ms mf
AB
(frfr)
ms
X C
(FrFr)
mf
ABC
(Frfr)
mf
Triple cross Hybrid
C X D
(frfr) (FrFr)
ms mf
CD
(Frfr)
mf
A X B
(frfr) (frfr)
ms mf
AB
(frfr)
ms
X
ABCD
1
(Frfr)
mf
1
(frfr)
ms
:
:
:
Double cross Hybrid
Simple hybrid with cms and
restoration
Maintainer line (B-line)
N, rfrf
N1
C1
xCMS line (A-line)
CMS, rfrf
N1
C1
Large amounts
of CMS line N1
C1
C2
x N2
Male line (C-line
N and RfRf
C1
Fertile F1 hybrid
CMS, Rfrf
Pearl Millet
CGMS
A1 Tift 23 A ( Most of the world hybrids contains
A1 Blood), Burton,1958
A2,A3 Not stable cytoplasm
A4 Derived from P.glacum subspecies monodii
Does not have effective restorer
Used in forage hybrid production
Cytoplasmic male-sterile
 Stamen (anther and filament) and pollen grains are
affected
 It is divided into:
a. Autoplasmic
CMS has arisen within a species as a result of spontaneous
mutational changes in the cytoplasm, most likely in the
mitochondrial genome
b. Alloplasmic
CMS has arisen from intergeneric, interpecific or occasionally
intraspecific crosses and where the male sterility can be
interpreted as being due to incompatibility or poor co-operation
between nuclear genome of one species and the organellar
genome another CMS can be a result of interspecific protoplast
fusion
Genetic Male Sterility
GMS is governed by two genes either recessive or dominant
genes(Kaul,1988)
One more dominant gene is associated with development of male
sterility in B.napus type by means of transgenic male sterility
Cytoplasmic Male Sterility
1.Raphanus or ogu system
2.Polima or pol system
3.Shiga-Thompson or nap system
4.Diplotaxis muralis or mur system
5.Tournefortii (tour) system
6. Moricandia arvensis or mori system
7.Chinese juncea or jun system
17 systems are available, only difference is the use of male sterile
cytoplasmic sources differs for each system
Nap system– B.napuus cross b/w winter & spring var.
pol system – B.napus var polima
mur system--Diplotaxis muralis x B.campestris cv Yukina
tour system– B.juncea collections
Ogu system:
First discovered in Japanese radish (Raphanus sativus) by Ogura, 1968
B.napus genome was transferred into the back round of R.sativus (mst)
through intergeneric crosses followed by back crossing with B.napus.
CMS seedling under low temperature showed chlorosis , because
chloroplast of R.sativus is sensitive to cold, it is governed by cp-DNA ,
but mst is governed by mt DNA.
Protoplast fusion of R.sativus with B.napus carried out to have normal
green plants with ogu CMS characterisitics
This system now has been used for developing alloplasmic male sterile line
in B.juncea and B.campestris.
Ogu system:
B.napaus
F1 interspecific cross
xRhapanus
sativus
F1 Sterile
G-Rs
C-Rs
G-Bn
N-Bn
1/2G-Rs
1/2G-Bn
C-Rs
mftmst
Doubling by colchince
Fertile amphidiploid
1/2G-Rs
1/2G-Bn
C-Rs
mst
Development of Male sterile Brassica napus from Rhapanus sativus
1/2G-Rs
1/2G-Bn
C-Rs
x G-Bn
N-Bn
G-Bn
C-Rs
B.napus
mst
BC3
Male sterile B.napus
mft
Development of Alloplasmic Male sterile Brassica campestris
x
N-Bc
B.campestris
F1 interspecific cross
xG-Bn
S-Rs
G-Bct
N-Bc
1/2G-Bn
1/2G-Bc
S-Rs
mftmst
G-BC
S-Rs
BC4
G-BcG-Bc
Male sterile B.napus
Chemical Male sterility
Enthrel – Brassica juncea
Zinc methy arsenate- B.napus
GA-B.oleracea var capitata
POTATO
Cytoplasm Nuclear genome Reference
S.acaule (4X) S.tuberosum Lamm,1953
S.chacoense(4X) S.tuberosum Rammanna and Hersmen(1974)
S.phureja(2x) S.tuberosum Magoon et al.,1958b
S.stoloniferum(4x) S.tuberosum Ross (1961)
S.Verrucosum(2X) S.tuberosum Abdalla (1970)
Inter-specific Hybridization
FW 450(Sodium B-Dichloro-iso-butyrate)
MH-30 (Maleic hydrazide)
Ethidium bromide
Chemical mutagens
Development of Male sterility
Genome transfer
S cytoplasm is in the genome of fr genes
Unreduced Gamete Production
S.tuberosum (2x) × S.tuberosum (4x)
(2x)
F1 (4x)
Protoplast Fusion
S cytoplasm is retained
Unreduced (2x) (2x)
F1 (4x)
Di haploid
S.tuberosum (4x) × S.phureja (4x)
(2x) (2x)
F1 (4x)
Anther culture
DiHaploid (2x)
Molecular Basis of Cytoplasmic Male sterility
Maintenance of ms determinant in populations. Maternal
inheritance (mitochondira) the male sterility determinant (red
cytoplasm) and the male fertility determinant (green cytoplasm)
are equally transmitted to the next generation
Male Sterility IN Cross Pollinated and Vegetable Crops

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Male Sterility IN Cross Pollinated and Vegetable Crops

  • 1. Male sterility in Cross Pollinated and Vegetative crops A.Manivannan Scientist (Genetics) DMR, New Delhi
  • 2.
  • 3. Male sterility in Sunflower (Helianthus spp) Genetic Male sterility (GMS) Complete male sterility ms1-ms5 = male sterility in sunflower recessive gene Two types of g-mst Type 1-gmst-Bloomington type Type 2-gmst-Modern type Cultivated Sunflower variety Karlik-68(Dwarf 68)- two recessive genes msi1,msi2(Stable and complete male sterile) Partial male sterility –p mst
  • 4. Source of cms through Interspecific Hybridization x H.annuusN1 C1 F1 interspecific cross H.grossessratus N1 C1 N1 C1 CMS- reccessive ms line X
  • 5. CGMS H.petiolaris × H.annuus Repeated backcross of H.annuus results in cms1 which is extensively used mst in hybrid seed production of sunflower all over the world H.giganteus× H.annuus Cms3( S cytoplasm source) H.annuus subspp lenticularis × H.annuus CV commander Indiana 1
  • 7.
  • 8. COTTON Genetic Male Sterility (GMS): 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. However, male sterility governed by recessive genes is used in practical plant breeding All three types of male sterility occurs (g mst,c mst,gc mst) in cotton sixteen different genes in tetraploid cottons (13 in G. hirsutum and 3 in G. barbadense) and two in G. arboreum have been identified for genetic male sterility. Sterility is conditioned by dominant alleles at five loci viz, MS4, MS7, MS10, MS11 and MS12 by recessive allele at other loci viz. msl, ms2, ms3, ms13, ms14 (Dong A), ms15 (Lang A) and ms16 (81 A). Two male sterile phenotypes viz. ms5ms6 and ms8ms9 are conditioned by duplicate recessive factors. G. hirsutum line Gregg (MS 399) from USA is the basic source of GMS possessing ms5 ms6 gene for male sterility.
  • 9. GMS
  • 10.
  • 11. CMS System In case of CMS, the originally discovered CMS sources involving G. arboreum and G. anomalum cytoplasmic systems having interaction with ms3 locus were not found effective or stable under different environments. The only stable and dependable CMS source under varied environment was developed through the utilization of G. harknessii. The complete genome of G.hirsutum was transferred into the G. harknessii cytoplasm. A single dominant gene ‘Rf’ from G.harknessii is essential for fertility restoration. Fertility enhancer factor 'E' for this CMS restorer system was obtained from a G.barbadense stock. The harknessii system is reported to contribute to good agronomic properties and attraction to honey bees.
  • 12. 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 G. aridum Skovt. × G. hirsutum (D4) G. trilobum × G. hirsutum CMS 8 (D-8) G. sturtianum × G. hirsutum CMS-C1 New sources of CGMS G. anomalum x G. thurberi Cg-mst
  • 13. Mutation G. arboreum, the first spontaneous male sterility mutant was identified in variety DS-5 Chemical based male sterility FW 450(Sodium B-Dichloro-iso-butyrate) MH-30 (Maleic hydrazide) Ethidium bromide Male sterility based hybrid Production GMS system. CPH2 (Suguna), First hybrid based on GMS released at CICR, RS, Coimbatore G. harknessii based cms with fertility restoration gene sources were used in developing the hybrid CAHH 468 (PKV Hy-3).
  • 14.
  • 15. T cytoplasm S cytoplasm C cytoplasm Discovered by Rogers(1944) M.T. Jenkins Beckett(1971) Developed from OP Mexican maize variety Gold june Teopod maize Brazilian Maize Male sterility Stable Unstable Stable HMT Susceptibility Yes No No Type of mst Sporophytic Gametophytic Sporophytic Fertility restoration Fr1,Fr2 - Fr4,Fr5, Fr6 CMS T cytoplasmic mitochondria, HM T pathotoxin causes the uncoupling of oxidative phosphorylation, inhibiton of oxogluterate oxidation and causes irreverisble swelling
  • 17. CGMS Reported by Rhoades (1931) while working with Peruvian source of maize cytoplasm Interspecific crosses Coix lacrymus-jobi X Z.mays Euchlaena mexican X Z. mays E.Perennis X Z.mays Chemcial mst GA, MH, Mendok, DPX 3778, Mo deficiency
  • 18. A X B (frfr) (FrFr) ms mf AB (Frfr) mf Method of Hybrid seed production Single cross Hybrid T- cytoplasm required two Fr genes C,S-cytoplasm requires one Fr genes Most of the T cytoplasm posses one Fr gene
  • 19. A X B (frfr) (frfr) ms mf AB (frfr) ms X C (FrFr) mf ABC (Frfr) mf Triple cross Hybrid
  • 20. C X D (frfr) (FrFr) ms mf CD (Frfr) mf A X B (frfr) (frfr) ms mf AB (frfr) ms X ABCD 1 (Frfr) mf 1 (frfr) ms : : : Double cross Hybrid
  • 21. Simple hybrid with cms and restoration Maintainer line (B-line) N, rfrf N1 C1 xCMS line (A-line) CMS, rfrf N1 C1 Large amounts of CMS line N1 C1 C2 x N2 Male line (C-line N and RfRf C1 Fertile F1 hybrid CMS, Rfrf
  • 23. CGMS A1 Tift 23 A ( Most of the world hybrids contains A1 Blood), Burton,1958 A2,A3 Not stable cytoplasm A4 Derived from P.glacum subspecies monodii Does not have effective restorer Used in forage hybrid production
  • 24.
  • 25. Cytoplasmic male-sterile  Stamen (anther and filament) and pollen grains are affected  It is divided into: a. Autoplasmic CMS has arisen within a species as a result of spontaneous mutational changes in the cytoplasm, most likely in the mitochondrial genome b. Alloplasmic CMS has arisen from intergeneric, interpecific or occasionally intraspecific crosses and where the male sterility can be interpreted as being due to incompatibility or poor co-operation between nuclear genome of one species and the organellar genome another CMS can be a result of interspecific protoplast fusion
  • 26. Genetic Male Sterility GMS is governed by two genes either recessive or dominant genes(Kaul,1988) One more dominant gene is associated with development of male sterility in B.napus type by means of transgenic male sterility
  • 27. Cytoplasmic Male Sterility 1.Raphanus or ogu system 2.Polima or pol system 3.Shiga-Thompson or nap system 4.Diplotaxis muralis or mur system 5.Tournefortii (tour) system 6. Moricandia arvensis or mori system 7.Chinese juncea or jun system 17 systems are available, only difference is the use of male sterile cytoplasmic sources differs for each system Nap system– B.napuus cross b/w winter & spring var. pol system – B.napus var polima mur system--Diplotaxis muralis x B.campestris cv Yukina tour system– B.juncea collections
  • 28. Ogu system: First discovered in Japanese radish (Raphanus sativus) by Ogura, 1968 B.napus genome was transferred into the back round of R.sativus (mst) through intergeneric crosses followed by back crossing with B.napus. CMS seedling under low temperature showed chlorosis , because chloroplast of R.sativus is sensitive to cold, it is governed by cp-DNA , but mst is governed by mt DNA. Protoplast fusion of R.sativus with B.napus carried out to have normal green plants with ogu CMS characterisitics This system now has been used for developing alloplasmic male sterile line in B.juncea and B.campestris. Ogu system:
  • 29. B.napaus F1 interspecific cross xRhapanus sativus F1 Sterile G-Rs C-Rs G-Bn N-Bn 1/2G-Rs 1/2G-Bn C-Rs mftmst Doubling by colchince Fertile amphidiploid 1/2G-Rs 1/2G-Bn C-Rs mst Development of Male sterile Brassica napus from Rhapanus sativus
  • 31. Development of Alloplasmic Male sterile Brassica campestris x N-Bc B.campestris F1 interspecific cross xG-Bn S-Rs G-Bct N-Bc 1/2G-Bn 1/2G-Bc S-Rs mftmst G-BC S-Rs BC4 G-BcG-Bc Male sterile B.napus
  • 32. Chemical Male sterility Enthrel – Brassica juncea Zinc methy arsenate- B.napus GA-B.oleracea var capitata
  • 34. Cytoplasm Nuclear genome Reference S.acaule (4X) S.tuberosum Lamm,1953 S.chacoense(4X) S.tuberosum Rammanna and Hersmen(1974) S.phureja(2x) S.tuberosum Magoon et al.,1958b S.stoloniferum(4x) S.tuberosum Ross (1961) S.Verrucosum(2X) S.tuberosum Abdalla (1970) Inter-specific Hybridization
  • 35. FW 450(Sodium B-Dichloro-iso-butyrate) MH-30 (Maleic hydrazide) Ethidium bromide Chemical mutagens
  • 36. Development of Male sterility Genome transfer S cytoplasm is in the genome of fr genes Unreduced Gamete Production S.tuberosum (2x) × S.tuberosum (4x) (2x) F1 (4x) Protoplast Fusion S cytoplasm is retained Unreduced (2x) (2x) F1 (4x)
  • 37. Di haploid S.tuberosum (4x) × S.phureja (4x) (2x) (2x) F1 (4x) Anther culture DiHaploid (2x)
  • 38. Molecular Basis of Cytoplasmic Male sterility
  • 39.
  • 40. Maintenance of ms determinant in populations. Maternal inheritance (mitochondira) the male sterility determinant (red cytoplasm) and the male fertility determinant (green cytoplasm) are equally transmitted to the next generation