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.
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.
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