Hort. Sci. (Prague), 2011, 38(1):27-34 | DOI: 10.17221/47/2010-HORTSCI

Efficiency of SSR markers for determining the origin of melon plantlets derived through unfertilized ovary culture

A.A. Malik, Li Cui, Shuxia Zhang, Jin-feng Chen
1 State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, China
2 Key Laboratory of Southern Vegetable Genetic Improvement, Nanjing Agricultural University, Nanjing, China

The effects of temperature pre-treatment, thidiazuron, naphthaleneacetic acid, and 6-benzylaminopurine on in vitro gynogenic plant production from un-pollinated melon (Cucumis melo L.) ovaries were investigated. Simple sequence repeat (SSR) marker analysis was conducted to identify the homozygous diploid individuals. The temperature pre-treatment (4°C) for 4 days increased embryo formation frequency (63.3%) significantly. Addition of thidiazuron (0.04 and 0.02 mg/l) in the induction medium significantly increased the number of responding ovaries (46.6%, 65.83%), respectively. The maximum number of plantlet regeneration (22.5%) was achieved by culturing the ovary derived embryos on Murashigue and Skoog medium (MS medium) supplement with 0.6 mg/l 6-benzylaminopurine. Spontaneous doubled haploids originated directly through embryogenesis were subjected to genetic analysis using SSR molecular marker with 23 primers pair for homozygosity. SSR markers with microsatellite CMGA172, confirmed that the alleles in the parental material were also present in the gynogenic plantlets, but amplified only two alleles as compared to four alleles of the heterozygous parent material at same locus. Therefore these regenerated plantlets were consider homozygous and produced through a process of gametophytic embryogenesis.

Keywords: Cucumis melo L.; in vitro gynogenesis; embryo formation; microsatellites markers

Published: March 31, 2011  Show citation

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Malik AA, Cui L, Zhang S, Chen J. Efficiency of SSR markers for determining the origin of melon plantlets derived through unfertilized ovary culture. Hort. Sci. (Prague). 2011;38(1):27-34. doi: 10.17221/47/2010-HORTSCI.
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    References

    1. Bajaj Y.P.S., 1990. In vitro production of haploids and their use in cell genetic and plant breeding. In: Bajaj Y.P.S. (ed.), Biotechnology in Agriculture and Forestry 12: Haploid in Crop Improvement I. Berlin, Springer-Verlag: 1-44. Go to original source...
    2. Bassam B.J., Caetano A.G., Gresshoff P.M., 1991. Fast and sensitive silver staining of DNA in polyacrylamide gels. Analytical Biochemistry, 196: 80-83. Go to original source... Go to PubMed...
    3. Bouvier L., Gúerif P., Djulbic M., Durel C., Chevreau E., Lespinasse Y., 2002. Chromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers. Euphytica, 123: 255-262. Go to original source...
    4. Cao Q.H., Chen J.F., 2003. Chromosome count using buds in vitro. Plant Physiology Communication, 39: 655-657.
    5. Chani E., Veilleux R.E., Boluarte M.T., 2000. Improved androgenesis of inter-specific potato and efficiency of SSR markers to identify homozygous regenerants. Plant Cell Tissue and Organ Culture, 60: 101-112. Go to original source...
    6. Chen Y., Hausner G., Kenaschuk E., Procunier D., Dribnenki P., Penner G., 1998. Identification of microspore-derived plants in anther culture of flax (Linum usitatissimum L.) using molecular markers. Plant Cell Reports, 18: 44-48. Go to original source...
    7. Cuny F., Dumas de Vaulx R., Longhi B., Siadous R., 1992. Analyse des plantes de melon (Cucumis melo L.) issues de croisements avec du pollen irradi a differentes doses. Agronomie, 12: 623-630. Go to original source...
    8. Danin-Poleg Y., Reis N., Tzuri G., Katzir N., 2001. Development and characterization of microsatellite markers in Cucumis. Theoretical and Applied Genetics, 102: 61-72. Go to original source...
    9. Diao W., Jia Y., Hui S., Zhang X., Lou Q., Chen J.F., 2009. Efficient embryo induction in cucumber ovary culture and homozygous identification of the regenerants using SSR markers. Scientia Horticulturae, 119: 246-251. Go to original source...
    10. Doyle J.J., Doyle J.L., 1990. Isolation of plant DNA from fresh tissue. Focus, 12: 13-15. Go to original source...
    11. Dryanovska O., Ilieva I., 1983. In vitro anther and ovule cultures in muskmelon (Cucumis melo L.). Proceedings of Bulgarian Academy of Sciences, 36: 1107-1110.
    12. Ficcadenti N., Sestili S., Annibali S., Marco M.D., Schiavi M., 1999. In vitro gynogenesis to induce haploid plants in melon (Cucumis melo L). Journal of Genetics and Breeding, 53: 255-257.
    13. Ficcadenti N., Veronese P., Sestili S., Crino P., Lucretti S., Schiavi M., Saccardo F., 1995. Influence of genotype on the induction of haploidy in Cucumis melo L. by using irradiated pollen. Journal of Genetics and Breeding, 49: 359-364.
    14. Gémesné Juhász A., Venczel G., Balogh P., 1997. Haploid plant induction in zucchini (Cucurbita pepo L. convar. giromontiina DUCH) and in cucumber (Cucumis sativus L.) lines through in vitro gynogenesis. Acta Horticulturae, 447: 623-626. Go to original source...
    15. Gürel S., Gürel E., Kaya Z., 2000. Doubled haploid plant production from unpollinated ovules of sugarbeet (Beta vulgaris L.). Plant Cell Reports, 19: 1155-1159. Go to original source... Go to PubMed...
    16. Hui S., Lou Q.F., Luo X.D., Wolukau J., Diao W., Qian C., Chen J.F., 2007. Regeneration of doubled haploid plants by androgenesis of cucumber (Cucumis sativus L.). Plant Cell Tissue and Organ culture, 90: 245-254. Go to original source...
    17. Kernan Z., Ferrie A.M.R., 2005. Microspore embryogenesis and the development of a double haploidy protocol for cow cockle (Saponaria vaccaria). Plant Cell Reports, 25: 274-280. Go to original source... Go to PubMed...
    18. Kumlehn J., Nitzsche W., 1995. Plant regeneration from isolated ovules of Italian ryegrass (Lolium multiflorum Lam.): effect of 2,4-dichlorophenoxyacetic acid and different cytokinins supplemented to the ovule culture medium. Plant Science, 111: 107-116. Go to original source...
    19. Lux H., Herrmann L., Wetzel C., 1990. Production of haploid sugar-beet (Beta vulgaris L.) by culturing unpollinated ovules. Plant Breeding, 104: 177-183. Go to original source...
    20. Morgante M., Olivieri A.M., 1993. PCR-amplified microsatellites as markers in plant genetics. Plant Journal, 3: 175-182. Go to original source...
    21. Mukhambetzhanov S.K., 1997. Culture of non-fertilized female gametophytes in vitro. Plant Cell Tissue and Organ Culture, 48: 111-119. Go to original source...
    22. Murashigue T., Skoog F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum, 15: 473-497. Go to original source...
    23. Panaia M., Senaratna T., Dixon K.W., Sivasithamparam A., 2004. The role of cytokinins and thidiazuron in the stimulation of somatic embryogenesis in key members of the Restionaceae. Australian Journal of Botany, 52: 257-265. Go to original source...
    24. Perera L., Perera V., Hocher J., Verdeil L., Yakandawala L., Weerakoon K., 2008. Use of SSR markers to determine the anther-derived homozygous lines in coconut. Plant Cell Report, 27: 1697-1703. Go to original source... Go to PubMed...
    25. Rashid M.A., Singh D.P., 2000. A manual on vegetable seed production in Bangladesh. AVRDC-USAID-Bangladesh Project Horticulture Research Centre, Bangladesh Agricultural Research Institute Joydebpur, Bangladesh, Gazipur: 89.
    26. Sauton A., 1989. Haploid gynogenesis in Cucumis sativus induced by irradiated pollen. Cucubit Genetics Cooperative Report, 12: 22-23.
    27. Sharma V.K., Hansch R., Mendel R.R., Schultze, J., 2005. Mature embryo axis-based high frequency somatic embryogenesis and plant regeneration from multiple cultivars of barley (Hordeum vulgare L.). Journal of Experimental Botany, 56: 1913-1922. Go to original source... Go to PubMed...
    28. Visser C., Qureshi J.A., Ravinder G., Saxena P.K., 1992. Morphology role of Thidiazuron: substitution of auxin and cytokinin requirements for the induction of somatic embryogenesis in geranium hypocotyl cultures. Plant Physiology, 99: 1704-1707. Go to original source... Go to PubMed...
    29. Wang M., Bergen S., Duijn B., 2000. Insights into a key developmental switch and its importance for efficient plant breeding. Plant Physiology, 124: 523-530. Go to original source... Go to PubMed...
    30. Zhang C.L., Chen D.F., Elliott M.C., Slater A., 2001. Thidiazuron-induced organogenesis and somatic embryogenesis in sugar beet (Beta vulgaris L.). In vitro Cellular and Developmental Biology, 37: 305-310. Go to original source...
    31. Zhao F.C., Nilanthi D., Yang Y.S., Wu H., 2006. Anther culture and haploid plant regeneration in purple coneflower (Echinacea purpurea L.). Plant Cell Tissue and Organ Culture, 86: 55-62. Go to original source...

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