Male sterility in vegetable crops by sunidhi mishra

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Definition : Absence or non functionality of pollen in plants(C.M.Rick,1944).
OR
Incapability of plants to produce or release functional pollen grains although the female gametes function normally.

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Male sterility in vegetable crops by sunidhi mishra

  1. 1. Utlilization of Male Sterility in Hybrid Seed Production of Vegetable Crops Presented by- Sunidhi Mishra Deptt. of Vegetable Science IGKV, Raipur
  2. 2. 2
  3. 3. Male sterility Definition : Absence or non functionality of pollen in plants(C.M.Rick,1944). OR Incapability of plants to produce or release functional pollen grains although the female gametes function normally.  J.K. Koelreuter (1763) observed anther abortion within species and species hybrids.  Jones and Emsweller (1936) identified the male sterility first in the onion and documented its maternal inheritance and use in hybrid seed production (Jones and Clarke, 1943).  More recently, work on male sterility has been reviewed by Frankel and Galun(1977) and Kaul(1988).
  4. 4. Hybrids  Hybrids are the F1 progenies obtained by crossing two genetically dissimilar parents.  Various mechanisms for developing hybrids are as follows-
  5. 5. Why male sterility for the development of hybrids ?  Production of large scale of F1 seeds.  Reduced cost of hybrid seed production.  Speedup the hybridization programme.  Commercial exploitation of hybrid vigour.
  6. 6. Features of Male Sterility  Prevents self pollination, permits cross pollination.  Leads to heterozygosity.  Female gametes function normally.  Male sterility is assayed through staining techniques (carmine, lactophenol or iodine), while detection of female sterility is by the absence of seeds.  In nature, occur due to spontaneous mutations.  Can be induced artificially.
  7. 7. Male sterility mechanisms used for development of hybrids in vegetables Mechanism Vegetables Remark Reference Nuclear male Sterility Tomato Monogenic recessive mutant was utilized to develop cost effective experimental crosses. Sawhney, 1997 Watermelon The utilization of monogenic recessive mutant was proposed. Zhang et al., 1994 Bottle gourd, Okra Male sterile plants were identified and utilized to develop experimental crosses. Dutta, 1983 Cytoplasmic male sterility Radish The combined use of both MS and SI mechanisms to enhance efficiency of hybrid seed production has been proposed. Cho et al., 1985 Functional male sterility Eggplant , Tomato A monogenic recessive mutant was identified and proposed for commercial utilization. Phatak & Jaworski, 1989 Transgenic male sterility Tomato, Cauliflower, etc. Many vegetables are at the edge of commercial utilization. Williams et al., 1997
  8. 8. Male sterility On Phenotypic basis On Genotypic basis Structural male sterility Sporogenous male sterility Functional male sterility Cytoplasmic male sterility Cytoplasmic genetic male sterility Genetic male sterility Kaul (1988) Classification of male sterility
  9. 9. Genetic / Nuclear male sterility  Reported in about 175 plant species (Kaul,1988).  Controlled by pair of recessive alleles “msms”, present in the nuclear compartment.  ms alleles arise spontaneously or may be artificially induced. Inheritance pattern Inheritance of GMS
  10. 10. Utilization of GMS in hybrid production x 1ms ms : 1Ms ms x Ms Ms A line C line F1 Hybrid ms ms A line Ms ms Ms Ms C line (removed before pollen shedding) Ms ms B line x
  11. 11. Environment-sensitive male sterility or “Two Line Hybrid Breeding”  These GMS line are conditional mutants.  Sensitive to temperature (TGMS) or photoperiod (PGMS).  Reported in several vegetable crops. Vegetables Mutants References Cabbage TGMS,PGMS Rundfeldt,1961 Brussels sprouts TGMS Niewhof,1968 Broccoli TGMS Dickson,1970 Pepper TGMS Daskalov,1972 Carrot TGMS Kaul,1988 Tomato TGMS Rick,1948;Sawhney,1983
  12. 12. Limitations of GMS  Because of more tedious maintenance process and non- availability of suitable marker gene among the vegetable crops, GMS has been utilized commercially only in chilli and muskmelon (Shifriss,1997 ; Kalloo et al.,1998).  Rouging of male fertile plants from the female line is time consuming and costly as a result of which the cost of hybrid seed is higher.  Use of temperature or photo-sensitive genetic male sterile lines (TGMS or PGMS) eliminates this problem.
  13. 13. Transgenic Genetic male sterility system Barnase-Barstar system
  14. 14. Utilization of barnase-barstar system
  15. 15. Cytoplasmic male sterility  It is the result of mutation in mitochondrial genome (mtDNA).  CMS is maternally inherited trait because mt genome is responsible for the expression of male sterility and the mitochondria are usually excluded from the pollen during fertilization.  Can be easily transferred from strain A to a given strain B.  CMS plants have also been developed in several vegetables through protoplast fusion ( Pelletier et al., 1995).  Three Ogura based improved CMS lines of cauliflower were developed following seven generations of backcrossing with snowball group.
  16. 16. Utilization of CMS in hybrid production
  17. 17. Cytoplasmic genetic male sterility  Case of cytoplasmic male sterility where nuclear gene, R for restoring fertility is known.  Fertility restorer gene should be present in homozygous Rf/Rf state i.e. either S Rf/Rf or N Rf/Rf where seed production is important.  The sterility factor is determined by the interaction of nuclear genes and cytoplasm but none of them singly can control sterility.
  18. 18. Utilization of CGMS in hybrid production
  19. 19. Limitations of CGMS  Non availability of CGMS in many crops and their wild relatives.  Need of fertility restorer allele in fruit producing vegetables.  Undesirable pleiotropic effect of sterile cytoplasm on horticultural qualities.  Breakdown of male sterility in particular environments.
  20. 20. List of gametocides found effective in vegetables Male gametocide Vegetables on which found effective Naphthalene Acetic Acid (NAA) Cucurbits Gibberellins Onion, Lettuce Maleic Hydrazide Tomato, Cucurbit, Onion FW450 Tomato, Ground nut, Sugar beet Ethrel Sugar beet Prasanth, et al., 2014
  21. 21. TOMATO
  22. 22. Description of different male sterile mutants in tomato  GMS: More than 55 male sterile (ms) alleles causing sporogenous, structural and functional sterility have been reported (Kaul,1988). Mutant Description Inheritance Governing genes Pollen sterile Pollen abortive Monogenic recessive (except MS-48, monogenic dominant ) ms series Stamenless Stamens absent Monogenic recessive sl-1, sl-2 Positional sterility Stigma exerted Monogenic recessive ps Functional sterility Anthers do not dehisce Monogenic recessive ps-2
  23. 23. CARROT Male fertile Male sterile
  24. 24. CGMS There are three types of male sterility in carrot :  Brown anther (ba ) Welch and Grimbal, 1947  Petalloid (pt) Thompson, 1962  Gummifer (1992)  Brown anther type male sterility is due to the interaction of “sa cytoplasm” with atleast two independent recessive nuclear genes.  According to Morelock (1974), the pt type of male sterility is due to interaction between “Sp cytoplasm” and two independent dominant genes (M1 and M2).
  25. 25. (a) Petaloidy CMS a) Normal (N cytoplasm) b) Brown anther CMS (Sa) c) Petaloid CMS (Sp) (a) Petaloid CMS (Sp) (b) Brown anther CMS (Sa)
  26. 26. Utilization  For hybrid production, petaloid steriles are employed more widely (Riggs,1987).  In India, at IARI, petaloid CGMS was transferred to nantes type and crossed it with the indigenous variety “Pusa yamdagini” to develop hybrid Pusa nayanjyoti in 2009.  Cytoplasmic male sterility system has been established for the first time in tropical carrot and tropical carrot hybrid “Pusa Vasudha” has been developed which is the first from any public sector institution.
  27. 27. Male sterility in cole vegetables  Genetic male sterility  Reported in most of the cole vegetables (Cole, 1957).  Mostly governed by a single recessive nuclear gene, ms.  Multiplication of ms seeds and existence of SI system is issue.  Cytoplasmic-Genetic male sterility  Firstly, developed by Pearson (1972) through interspecific hybidization between wild mustard and broccoli.  Sources of CMS  Sterile ‘Anand’ cytoplasm from B.rapa has been transferred to cole crops through protoplast fusion .
  28. 28.  Ogura cytoplasm of Raphanus is a source of sterile (Ogura 1968).  Ogura cytoplasm was transferred in broccoli (McCollum,1981) ,cauliflower (Hoser-Krause and Antosik,1987), Brussels sprout (Bannerot et al., 1974) and in cabbage (McCollum,1981, 1988).  One such CMS system 'tour' which is derived from Brassica tournefortii, induces additional floral abnormalities and causes chlorosis in Brassica spp.(Arumugam et al. 1996). Ogura CMS system (Intergeneric crosses)
  29. 29. Chilli
  30. 30. Chilli  First documented by Martin and Grawford (1951) and first ms plant was isolated from an Indian accession (Peterson,1958).  GMS  The mc-509 was renamed as ms-10.  This ms-509 line (bell pepper type) was introduced in India at PAU and was introgressed in 3 chilli genotypes, Viz., MS-12, MS-13 & MS-41 (Singh and Kaur,1986).  MS-12 line has been developed by transferring ms-10 gene into cultivar ‘Punjab Lal’ through backcrossing (Singh and Kaur,1986).  Using MS-12 line three chilli hybrid CH-1, CH-3 and CH-27 has been released by PAU.
  31. 31.  GMS line, “ACMS2”, having gene (acms2acms2) is reported .  The ms-3 line introduced from Hungary is maintained at AVRDC, Taiwan(Berke,1999).  CGMS in chilli  First reported by Peterson(1958) in an introduction from India (PI-164835) .  Chilli CGMS lines (CCA-4261) introduced at the IIVR from AVRDC are utilized to produce hybrid- Kashi Surkh (CCH-2) .  IIHR, Bangalore has also developed CGMS based hybrids i.e. Arka Meghna (MSH-172), Arka Harita (MSH-149) and Arka Sweta (MSH-96).
  32. 32. Male fertile flower Male sterile flower ONION
  33. 33. Male fertile Male sterile
  34. 34.  CMS –  First ms plant (13-53) was reported in the progenies of cultivar Italian Red (Jones and Emsweller,1936)  It is due to 2 recessive genes ms1and ms2.  Two types of sterile cytoplasm , viz., S and T are reported.  S-cytoplasm is exploited most widely (Pelletier et al., 1995).  Female to male ratio for hybrid production is 4:1 or 8:2  National Released Hybrids  Arka Kirtiman & Arka Lalima (IIHR, Bangalore)  Hybrid-63 and Hybrid-35 (IARI, New Delhi)
  35. 35. Muskmelon  Genetic male sterility Male sterile alleles identified in muskmelon are-  The ms-1 line has been used in India to develop first commercial hybrid Punjab hybrid-1 and Punjab Anmol. in vegetable crops through male sterility at PAU.  Kumar et al., 2008, reported that the lines ms-1 and ms-2 are phenotypicaly unstable and this will reduce the genetic purity of hybrid seed. It is safer to exploit ms-3, ms-4 and ms-5 genes for developing genetically pure hybrids. Male sterile alleles Reported by ms-1 Bohn and Whitaker (1949) ms-2 Bohn and Principe, 1962 ms-3 McCreight and Elmstrom, 1984 ms-4 Pilrat,1990 ms-5 Lecouviour et al., 1990
  36. 36. Time(minutes) required for crossing 50 flower buds on male fertile ‘Ms33 IPA’ (MF) and male sterile ‘ms33IPA’ (MS) plants in tomato. Ludhiana Dhaliwal and Cheema,2008 Worker Time (minutes) Time saved in GMS over MF (%) Emasculation Pollinatio n on MF Emasculatio n + pollination on MF Pollinatio n on GMS 1 22.0 44.0 66.0 37.7 42.9 2 26.1 34.9 60.5 26.3 56.5 3 38.7 45.1 83.8 33.3 60.3 4 37.3 43.8 75.5 34.2 54.7 5 32.8 41.1 73.9 32.7 55.8 Mean 30.7 41.7 71.9 32.8 54.4 CD at P=0.05 1.52 1.60 1.95 1.10 -
  37. 37. Per cent fruit set on male sterile (GMS lines) plants in chilli. GMS line Mean percent fruit set on GMS line Natural out crossing Hand pollination ACMS2-1-1-1 32.64 13.82 ACMS2-1-1-4 30.22 14.08 ACMS2-5-1-1 35.99 14.82 ACMS2-5-1-5 34.39 14.00 ACMS2-6-1-1 30.68 15.34 ACMS2-6-1-3 32.82 14.60 S. Em - 0.35 Average of mean percent fruit set on GMS lines 32.79 14.44 Anand Patel et al. (2001)
  38. 38. Fruit characteristics of different crosses in tomato. Crosses Types of sterility Polar diameter (cm) Equatorial diameter (cm) Number of locule Pericarp thickness (mm) Total Soluble salt (%) Fruit weight (g) ms33 IPA x Ms33 IPA Pollen abortive 5.13 4.56 2.33 6.33 4.60 59.00 ms2 IPA x Ms33 IPA Pollen abortive 5.09 4.43 2.00 6.33 4.75 62.33 ps2 L 3841 x Ms33 IPA Functional 4.26 4.73 4.66 6.00 4.16 61.33 ps2 NS 101 x Ms33 IPA Functional 4.40 4.36 3.00 6.00 4.60 59.33 ps2 San Pedro x Ms33 IPA Functional 5.53 6.23 3.83 6.66 4.63 112.66 ps2 UC 82-B x Ms33 IPA Functional 5.15 4.70 2.10 6.30 4.80 67.00 C.D. at P=0.05 - 0.34 0.29 2.82 0.82 0.25 7.19 Ludhiana Dhaliwal and Cheema (2008) Male sterile lines: Six x Male fertile (Ms33 IPA)
  39. 39. Limitations of Male Sterility system  Difficult to identify line with GMS.  Unsatisfactory or poor pollination.  Unsatisfactory restoration of fertility.  Adverse effect of sterile cytoplasm on yield in some cases.  Break down of male sterility because of some reasons like, certain environmental conditions which leads to some pollen production by the male sterile lines and cytoplasm contribution (though small) by the sperm in some cases.  Modifiers or modifying genes may affect cytoplasmic male sterility.
  40. 40. Conclusion….
  41. 41. Thank you

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