An efficient in vitro propagation protocol via shoot multiplication was developed for Rauvolfia hookeri, a rare medicinal species endemic to the southern Western Ghats of India. Shoot tip explants cultured on half strength Murashige and Skoog medium supplemented with various cytokinins (6-benzylaminopurine, 6-furfurylaminopurine and Thidiazuron) either alone or in combinations produced multiple shoots. When either cytokinin was used alone, 6- enzylaminopurine

was found nearly twice more successful than 6-urfurylaminopurine. The highest shoot proliferation was obtained when 6-benzylaminopurine and 6-furfurylaminopurine was used at 22.2 µM and 4.64 µM respectively. Thidiazuron gave the lowest response for shoot proliferation. The effect of indole-3-butyric acid and naphthalene acetic acid was evaluated for in vitro root induction. Rhizogenesis of excised shoots was of the shoots was readily achieved on half strength Murashige and Skoog medium containing various concentrations of indole-3-butyric acid and naphthale ne acetic acid. Indole-3- butyric acid was found to be more effective than naphthalene acetic acid and resulted in the highest frequency of shoots that rooted (86.5%) and mean number of roots per shoot (3.66) when used at 7.38 µM concentration. The micropropagated plants were hardened and transferred to green house condition wherein 70% plants established and were morphologically similar to mother plant. Genetic stability of regenerated plants has been checked by Random Amplified Polymorphic DNA using ten selected decamer primers.