Protoplast isolation and culture of Aranda hybrids
1988
Koh, M.C. | Goh, C.J. | Loh, C.S. (National University of Singapore (Singapore). Dept. of Botany)
Protoplasts were successfully isolated from leaf explants of Aranda Tay Swee Eng and A. Noorah Alsagoff. Various factors which affect protoplast isolation and culture were studied. Conditions found to be optimal for the isolation of viable protoplasts were: a) inclusion of driselase in the enzyme mixture, b) exclusion of calcium chloride pretreatment, c) use of younger leaves (1-3cm), d) an incubation temperature of 30 degree centigrade, e) a 10-hour and 7-hour incubation period for A. Tay Swee Eng and A. Noorah Alsagoff respectively, f) a 16L:8D photoperiod for donor plantlets, g) agitation on a gyrotory shaker at 40 rpm and h) 0.4M sucrose as osmoticum in the enzyme mixture. Plant growth regulators were found to be necessary for protoplast viability. After two weeks in culture, about 53 % of the protoplasts cultured in media supplemented with 2,4-D were still viable. Protoplast viability was also affected by various macro-nutrients as well as photoperiodic conditioning of the donor plantlets
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