In Vitro Regeneration of Hemidesmus indicus L. R. Br an important Endangered medicinal plant

Hemidesmus indicus belongs to the family Asclepiadaceae, and it is exceptionally demanded by traditional healers for the ailment of various diseases. The over exploitation of Hemidesmus indicus placed it under the endangered medicinal plant, and it required to develop in vitro culture for conservation and maintain the sustainable demand of plant. In the present study, we planned to develop rapid and efficient protocol for large-scale propagation of Hemidesmus indicus through in vitro culture. Cotyledons from in vitro germinated seeds  were used as initial explants, inoculated in MS medium supplemented with various cytokinins, BAP(0.5-2.0 mg/l)  and kn (0.5-1.0 mg/l)) in combination with auxin, NAA (0.5-1.0 mg/l) and IAA(0.1-0.5 mg/l). The optimal response of shoot initiation (80%) with average number of shoots 6.8 ± 0.10 ( mean length 6.4 ± 0.13) was observed in the medium (I-9) containing 2.0 mg/l BAP and 0.5 mg/l NAA within 4 week. The maximum shoot multiplication (84%) with average number of shoots 24.9 ± 0.15, was observed on MS medium M5 containing BAP (1.0 mg/l) in combination with Kn (0.5 mg/l). regenerated shoots were excised aseptically and implanted on MS half and full strength medium fortified with various concentrations of IBA, IAA and NAA for root formation. Full strength MS medium (R9) having 0.5 mg/l IAA was found better with 60% root formation after 16-18 days. The rooted plantlets were successfully acclimatized in pots containing sterilized soil and sand mixture (3:1) with 95% survival rate in the field conditions.