Phyla nodiflora (L.) is one of the highly demanded medicinal crops. Nodal explants of Phyla nodiflora (L.) were cultured on Murashigi and Skoog (MS) medium supplemented with different combination and concentration of cytokinins for multiple shoot regeneration. Maximum number of shoot regeneration occurred in1.0 mg/l BAP Compared to kinetin. Among twenty five different combinations of BAP & NAA, BAP was found to be the best for producing creamish green fast growing and friable callus. This protocol can be used for continuous and rapid multiplication of Phyla nodiflora. Moreover, plant secondary products are often produced only in small quantities in most of the plant species. It is not always feasible to isolate secondary compounds from intact plants. Therefore, plant cell and tissue culture techniques can be an alternative approach to maintain sustainability supply of plant materials for producing bioactive compounds continuously under artificially controlled condition.