In vitro propagation of elite clone of Ephedra gerardiana: an endangered medicinal plant

Ephedra gerardiana Wall. is an endangered and important medicinal plant, being used in various herbal formulations. Present report describes the development of a micropropagation protocol using BAP and Kn followed by assessment of the genetic integrity of in vitro raised plants. Most responsive media was MS Basal Medium supplemented with 15 µM Kn and 5 µM BAP showing the best results in terms of both percentage response and maximum number of shoot buds. The shoots rooted best onto ¼ MS containing 20 µM IBA. Hardening was followed by transplantation and finally field establishment where 76.8% of plants survived in nature. Genetic integrity of in vitro raised plants was also carried out by using 27 arbitrary decamer and 18 seventeen mer RAPD primers. A total of 16 primers exhibited monomorphism in the mother plant, callus derived plants and direct regenerated plants of E. gerardiana. Besides this, 6 primers showed amplification in six samples and four primers in five samples out of seven. The results also illustrate that RAPD markers indicated the absence of polymorphism in a total of 35 loci in the mother plant and regenerants. The banding pattern was constant within all in vitro raised plantlets and mother plant.