According to the WHO about 80% population of the world depend on medicinal plants for remedies. Medicinal plants are exploited for therapeutic medicine and have been used by Ayurveda, Unani and Siddha. In Ayurveda, Charak and Sushruta described various plants along with their medicinal values. Portulaca grandiflora, is a medicinal ornamental plant in the family Portulacaceae, Portulaca has many medicinal properties. It is used for the treatment of cirrhosis of the liver, pharyngeal pain and swelling. Leaves used for scurvy. Fresh juice of leaves and stems applied to snake and insect bites, burns, scalds, and eczema. In the present study micropropagation of Portulaca grandiflora was done using nodal explants, inoculated on MS medium containing kinetin (1.0-4.0mg/l). Axillary bud... proliferation was observed within 05 days. After 3 weeks regenerated shoots shown fresh growth with 3-4 new nodes on each concentration of kinetin supplemented Murashige and Skoog medium. No callus initiation was observed on any of the Kinetin supplemented media. Out of the four concentration 2.0 mg/l Kinetin shown maximum percentage of Axillary bud proliferation i.e. 60%, whereas higher concentration of Kinetin shown stunted growth. Regenerated shoots were transferred into Murashige and Skoog media supplemented with 1.0- 4.0mg/Indole Butyric Acid (IBA) for root initiation. Root initiation was observed after 5 days of subculturing. Maximum percentage of root initiation and plantlet regeneration were recorded in 2.0mg/l IBA i.e. 90%, whereas minimum percentage observed on 4.0mg/l i.e 40%. Regenerated Plantlets were transferred to green house and finally to the field.