An efficient in vitro protocol has been developed for the multiplication of shoots and conservation of a rare succulent medicinal plant Caralluma stalagmifera var. longipetala growing wildly in Karnataka State. Proliferation of multiple shoots was achieved on Murashige and Skoog’s (MS) medium supplemented with various concentrations of 6-benzyladenine (BA), Kinetin (Kn), indole acetic acid (IAA), α- naphthalene acetic acid (NAA) and indole-3-butyric acid (IBA) alone or with various combinations from the nodal explants. The nodal explants cultured on medium containing BAP (2.0 mg/L) along with 0.5 mg/L Kn and 0.3 mg/L IAA produced the highest number of shoot sprouting (2.60±0.16) and maximum shoot length (3.96±0.20). The considerable frequency of callus induction and embryogenesis was noticed both in 1.0 mg/L NAA and 0.5-2 mg/L, 2, 4-D. The calli transferred to shoot induction medium containing the combination of hormones BAP (1.0 mg/L) plus IAA (0.2 mg/L) and NAA (0.1 mg/L) successfully regenerated in vitro shootlets. The in vitro rooting was achieved from both direct shoots regenerated from nodal explants and callus derived shootlets with NAA (0.2 mg/L). The in vitro rooted plantlets were successfully acclimatized (75%) in the greenhouse and gradually transferred to open field conditions. Key word: Micro propagation, medicinal plant, Caralluma stalagmifera var. longipetala.