An efficient and improved plant regeneration protocol has been developed from nodal explants of Ocimum canum Sims., a medicinally important herbaceous plant species belonging to the family Lamiaceae. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l -1 N 6 -benzyladenine (BA) showed maximum percentage of shoot multiplication (91.1 %) with 5.32 shoots per explant. Rooting of shoots occurred on 1/2 MS medium supplemented with 1.0 mg l -1 indole-3-butyric acid (IBA). Well-developed plantlets, transferred to plastic pots containing sand, soil and compost (2:2:1) showed 80.9 % survival rate under field condition. The genetic fidelity of in vitro raised field-grown plants to the donor plant was ascertained from histological, cytological and two PCR based molecular markers, i.e., random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR). Histological study showed direct organogenesis on epidermal and sub epidermal layer, where as cytological studies showed no visible anomalies within its chromosome compliments. Both RAPD and ISSR analysis revealed high degree of monomorphism, similar to those of the mother plants. This method of in vitro clonal propagation of O. canum Sims. may, therefore, could be an avenue of its sustainable commercial exploitation and conservation.