Abstract: Rubia cordifolia (Rubiaceae), a rare and highly medicinal plant of Western ghats of India, needs to be conserved as its primary gene pool is threatened by deforestation and extensive exploitation by pharmaceutical industries which has resulted in disappearance of natural habitats of this species. Therefore, rapid multiplication and conservation of R. cordifolia is of paramount importance. An in vitro multiplication protocol for this plant has been developed by using axillary bud culture. MS basal medium supplemented with a synthetic cytokinin, thidiazuron (1 mg L-1) induced highest number of shoots per tube (12.67) with 85% of response, as compared with other cytokinins. Rooting of micropropagated shoots were achieved on half strength MS basal medium supplemented with NAA (2 mg L-1). Hardening of plants was accomplished by transfer of rooted plants to a mixture of soil, sand and compost (1: 2: 1) and then plantlets were transferred subsequently to the field. Using this axillary bud technology from single bud, more than 1000 plants were raised. This has proven to be the best tool for conservation of R. cordifolia and its rapid multiplication as well.