Plant Regeneration Through Indirect Somatic Embryogenesis in Coelogyne Cristata Orchid

A newly efficient protocol has been established for high frequency somatic embryogenesis through callus culture of Coelogyne cristata. The best frequency of callusing was obtained from leaf segments (3-5 mm) cultured on the MS medium supplemented with 2 mg?L–1 2,4-D and 2 mg?L–1 BA combination. A negative effect of coconut water was ob-served on the callus induction medium. When callus lines no. 4, 6 and 8 induced from leaf segments were sub-cultured separately on ½ MS and MS media containing AC (1-3 g?L–1), formation of somatic embryos was found. However, percentages of embryo formation and the number of embryos per explants were strongly affected by media and callus lines used. The effect of 1/2 MS media is definitely better than MS medium for somatic embryogenesis from the selected lines of leaf derived callus. Among the callus lines, line no. 4 is the best for somatic embryogenesis followed by line no. 6 and 8. The somatic embryos converted into healthy plants with well developed shoots on the same media. The plant-lets were transferred to 1/2 MS medium containing 1 g?L–1 AC for plant regeneration until 8 weeks of culture and successfully acclimatized in the greenhouse.