Studies have been conducted using Thevetia peruviana (Apocynaceae) leaf explants to evaluate the effect of auxin 2.4-D on varying concentrations and combinations of cytokinins (KIN, BA) for the induction of somatic embryos, as the taxon is a rich treasure of many natural cardiac glycosides. Soft, friable creamy yellow calli was obtained in 2, 4-D + KIN combination, where as it was creamy white and granular in 2,4D + BA. Results indicated that most of the tested concentrations were effective, but lower concentrations of 2, 4-D + KIN (0.5 - 1.0 mg/L-1) were more conducive than higher (2.0 - 3.0 mg/L-1) combinations for the induction of somatic embryos. Among the cytokinins, KIN is more efficient than BA. An increase in KIN concentration from 0.5 to 3.0 mg/ L-1, keeping 2.4-D constant (1.0 mg/ L-1) resulted in greater callus biomass with lesser frequency of somatic embryos, and the frequency of embryogenic masses decreased as the KIN concentration increased. Mature detached bipolar embryos with well-developed cotyledons and radicles developed into fully matured plantlets in 68% of culture tubes successfully in full strength MS medium having IBA + BA (0.1 - 0.5 mg/L-1) for regeneration. In order to prevent natural erosion of genetic resources, this successful regeneration protocol is useful for large scale multiplication within a short period of time and to minimize the impact of over exploitation by therapeutic industrialists.