Commiphora wightii (Arn.) Bhandari, is a medicinal important desert species of the family Burseraceae. It is a well known for its valuable active principle found in its oleo-gum-resin (guggulsterone E and Z), which are used in drugs preparation for lowering the cholesterol level in human body. Due to its overexploitation, poor natural regeneration this valuable plant is on the verge of extinction and thus a IUCN Red listed species. In the present study we report development of an efficient micropropagation protocol from cotyledonary node of Commiphora wightii. Cotyledonary nodes were used as an explants and multiple microshoots were obtained on Murashige & Skoog (MS) medium supplemented with 2.68 µM a-Naphthalene acetic acid (NAA) and 4.44 µM 6-Benzylamino purine (BAP) and on 2.68 µM NAA; 4.44 µM BAP with additives (glutamine 684.2 µM; thiamine 29.65 µM; activated charcoal 0.3%) and various other hormonal combinations. Elongation of microshoot was significantly observed on the 2.46 µM Indole-3-butyric acid (IBA) and 2.22 µM BAP supplemented MS medium. Efficient rooting was obtained on pretreated microshoot (4.92 µM IBA for 24 hours) and followed by transfer to White’s medium without Plant Growth Regulators (PGR) and high concentration of activated charcoal (AC). Rooted micro-shoots were transferred to vermiculite and wetted with Hoagland’s solution for primary hardening for 4-5 weeks and then finally transferred to plastic cups containing vermiculite, placed in mist chamber. Plantlets were transferred to soil: FYM 1:1 containing poly-bags, then to green shade house for complete acclimatization and finally transplanted to the experimental field