In vitro REGENERATION FROM LEAF SEGMENTS OF ENDANGERED MEDICINAL PLANT Withania coagulans (STOCKS) DUNAL

Withania coagulans is an important medicinal plant which has become critically endangered over the years owing to extremely poor germination rate, slow propagation in nature, unrestricted collection from wildstands for traditional as well as medicinal purposes and destruction of habitat. The present study was undertaken to develop in vitro regeneration protocol for mass propagation as well as conservation of critically endangered medicinal plant Withania coagulans without causing damage to mother plant. In the study leaf segments obtained from mature plant of W. coagulans were surface sterilized and aseptically cultured onto MS medium fortified with different hormones (NAA, Kn, BAP) either alone as well as in combination and development of callus was achieved from leaf segments cultured onto MS+NAA+BAP and MS+NAA+Kn. Subculture of callus onto MS+IAA (2-20µM) resulted in regeneration of shoots from callus. Formation of hairy root was also achieved onto IAA supplemented medium. In vitro regenerated shoots were rooted onto full as well as half strength MS medium enriched with varying concentration of either IAA or IBA. In vitro rooting from regenerated shoots was achieved onto full and half strength MS medium containing IBA and half strength MS medium containing IAA. Maximum rooting in about 88.4% cultures was obtained onto ½ MS+20µM IBA followed by 85.2% in vitro rooting onto ½ MS+20µM IAA. About 64.5% regenerated plants were successfully acclimatized to natural conditions.