Abstract
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.
Highlights
Coelogyne cristata which belongs to the family Orchidaceae has high ornamental value as a cut flower
This study presents optimal concentrations of 2,4-D and BA for callus formation and high formation of somatic embryos from the culture of leaf derived callus in Coelogyne cristata using different kinds of media and activated charcoal (AC)
The germinated plantlets about 1.0 cm in height were transferred to Murashige and Skoog (MS) basal media supplemented with NAA 1.0 mg·L–1 and BA 2.0 mg·L–1 for plant growth
Summary
Coelogyne cristata which belongs to the family Orchidaceae has high ornamental value as a cut flower. It produces its graceful racemes of white flowers with yellow splotches on the throat, and has a long lasting and fragrant scent. There is no report on plant regeneration through somatic embryogenesis in C. cristata. Success of somatic embryogenesis via callus culture is interesting in plant regeneration of orchids. This study presents optimal concentrations of 2,4-D and BA for callus formation and high formation of somatic embryos from the culture of leaf derived callus in Coelogyne cristata using different kinds of media and AC
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