Multiple shoots were regenerated from cotyledonary node, shoot tip explants derived callus by one stage process where caulogenesis occurred on the primary callus. However cotyledons, leaves and root segments gave rise to adventitious shoots from callus by two stage method of indirect organogenesis. High frequency of shoot differentiation was observed from cotyledon and leaf on MS medium fortified with BA (8.9 to 13.3 μM) in combination with NAA (0.54 μM) compared to other explants. Further, when elongated shoots were subjected to rooting, we observed shoot necrosis consistent to our previous findings. Similarly, we could overcome this problem by supplementing calcium to MS rooting medium. In vitro rooted plantlets were successfully transferred to soil, exhibiting a normal development. Altogether, our protocol provides a rapid technique for mass multiplication as well for ex situ conservation of this endemic medicinal plant.
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