In vitro culture of Cucumis sativus L. XVIII. Plants from protoplasts through direct somatic embryogenesis |
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Authors: | W. Burza S. Malepszy |
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Affiliation: | (1) Department of Plant Genetics and Breeding in Horticulture, Warsaw Agricultural University, Nowoursynowska 166, 02-766 Warszawa, Poland |
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Abstract: | A procedure is described for the isolation and culture of protoplasts from embryogenic callus (gel-like callus — GLC) and embryogenic suspension cultures (ESC) of Cucumis sativus c.v. Borszczagowski. Maximal protoplast yields from GLC and ESC were 5×106 and 1×107 protoplasts/g tissue respectively. They were obtained following 14–16 h digestion with 1.2% Cellulase Onozuka R-10, 1.2% Macerozyme R-10 and 0.3% Driselase. At a plating density of 2×105 / ml, first divisions occurred in 4–5 days and 7–8 days in ESC-and GLC-derived protoplasts respectively. The highest percentage of direct embryogenesis (over 80%) was observed with ESC. It was possible to obtain approximately 5000 embryo structures / g tissue. Some embryos converted into plants after 6 weeks, but most of them after 2 months of culture. ESC-derived plants, when transferred into the glasshouse, bloomed normally, and set seeds.Abbreviations CMS Murashige & Skoog (1962) medium for cucumber - GLC gel-like callus - ESC established embryogenic suspension culture - 2,4-d 2,4-dichlorophenoxyacetic acid |
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Keywords: | cucumber direct somatic embryogenesis established embryogenic suspension culture gel-like callus regeneration |
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