Plant Regeneration from Callus Culture of Clematis gouriana Roxb. – A Rare Medicinal Plant

摘要

An in vitro regeneration protocol through stem callus culture has been standardized for the medicinal climber Clematis gouriana. The explant induced callus on MS-medium supplemented with 0.5 to 1.5 mg l-1 BAP and 0.1 to 0.5 mg l-1 NAA. The optimized callus induction occurred at the concentration of 1.0 mg l-1 BAP and 0.3 mg l-1 NAA. After initiation of callus, it was immediately transferred to MS medium containing 4.0 mg l-1 FAP and 0.5 mg l-1 indole-3-butyric acid (IBA). Upon longer incubation for about 2-6 weeks on the same culture medium initiation of shoot buds from the callus mass was observed and regeneration of plantlets with higher frequency (mean of 11.1 ± 0.23 shoots per explants) was noted. The microshoots rooted well on MS basal medium without growth regulators, as well as on medium supplemented with 0.5 mg l-1 IBA. Regenerated shoots formed complete plantlets on medium containing 0.5 mg l-1 IBA, and mature plants were established, acclimatized, and thrived in greenhouse conditions. The regeneration protocol developed in this study provides a basis for germplasm conservation and for further investigation of bioactive constituents of this medicinal plant.