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[9000507.] رقم البحث : 9000507 - |
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Studies on some factors affecting anther culture in six cucumber genotypes / |
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Agricultural Research Journal Suez Canal University / |
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F.H. Mohamed. |
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M. W. M. Elwan. |
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M. M. Abd-El Salam |
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N. A. El-Maghawry |
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Cucumis sativus L., anther culture, haploid plants, cold treatment, medium components. |
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Three experiments were conducted to develop an efficient cucumber (Cucumis sativus L., 2n= 2x=14) anther culture protocol. The first experiment was conducted to study the effect of genotypes, medium types and stage of male flower development from which anther will be cultured. Difference among cucumber genotypes was significant for their effect on frequency of anther culture development into callus, but callus size was not affected. Medium type (MS) positively affected the percentage of anther development into callus than B5 medium. Anthers from closed flower buds were significantly better in callus formation than those taken from open flowers. The best combination which affected anther growth was achieved using anther derived from closed flower buds of the genotypes Garawani, L82-57 or L88-83 when grown on solid MS medium +2,4-D at 2.0 mg/l+ BA at 1.0 mg/l. As indicated from histological examination, none of the tested treatments showed callus regeneration. The second experiment examined the effect of 24 combinations of glutamine and growth regulators types and concentration on callus growth from cold or non-cold treated anther of 6 cucumber genotypes. MS medium amended with 1.0 mg/l 2,4-D+0.5 mg/l BA+ 1.0 mg/l kin was the most effective on callus growth scale. Cold pre-treatment of anther at 4 ?C for 48 hr resulted in more vigorous callus than non-cold. When callus was examined under microscope, embryo-like callus was observed in L 88-83 with the culture of cold-treated anther on MS medium + 0.5 mg/l 2,4-D + 0.5 mg/l BA, while shoot primordia appeared in medium containing 1.0 mg/l 2.4-D + 1.0 mg/l BA + 1.0 mg/l kin and 50 mg/l glutamine. Very visible shoot or root initials were observed in non-cold treated anther culture of the genotype WI 1701G in a medium devoid of glutamine, indicating difference among cucumber genotypes in their response to medium composition (PGR and glutamine) or thermal treatment. These results were confirmed by histological analysis. In the third experiment anther-derived callus from Exp.2 were sub-cultured into MS medium amended with 3 concentrations of ABA and two sucrose levels, to examine further differention of callus into regenerants. Microscopic examination of the resulted culture indicated shoot bud regeneration in a medium with 0.3 mg/l ABA+ 9% sucrose, while hairy roots were visible from sub culturing callus into 0.2 mg/L ABA+3% sucrose. Embryogenic like callus with side shoots was formed on MS medium + 0.1 mg/l ABA + 9% sucrose. Histological examination provided clear evidence of shoot bud formation in the above medium. |
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