Present study describes micro-propagation of traditionally important medicinal plant in northern interior part of Karnataka, Caralluma adscendens var. fimbriata, for its conservation. Results showed that the survival ability of regenerated plants was 84 per cent. Proliferation of C. adscendens var. fimbriata shoots was achieved on medium supplemented with various concentrations of different hormones. The effective combination of 6-Benzylaminopurine (BA) 2.0 mg/l along with 0.5 mg/l kinetin (Kn) and 0.2 mg/l Indole-3-acetic acid (IBA) produced highest number of shoot sprouting frequency (91%) with a mean of 2.60 ± 0.17 shoot number per explants and a mean shoot length of 3.88 ± 0.20 cm from the mature nodal explants. A good frequency of callus was induced in 1.5 to 2.0 mg/l of 1-Naphthaleneacetic acid (NAA) and in 2.0 mg/l of 2,4-dichlorophenoxy acetic acid (2, 4-D) were transferred to root induction media. The NAA showed positive response of rooting when compared with IAA and IBA in the present study. The best root and shoot sprouted plants were successfully acclimatized in the greenhouse and transferred to open field conditions. This micropropagation protocol can be successfully used for large-scale multiplication and conservation of germplasm of this threatened plant.