The need of MS salts for shoot sprouting and proliferation shows the high salt requirement for the growth of Gymnema sylvestre. Influence of plant growth regulators, IAA, BA, 2,4-D and kinetin on shoot sprouting was investigated. Synergistic effect of Vitamin B2 was studied. To overcome phenolic exudation and the effects of antioxidants, the effect of activated charcoal, citric acid and ascorbic acid were investigated. Incorporation of citric acid (100 mg L-1) to the medium prevented phenolic exudation and increased the production of healthy normal shoots and shoot bud differentiation in Gymnema sylvestre. There was a considerable improvement in rooting as about 53% shoots could be induced to root on 1/2 MS medium within 45 days with a fairly good length and number of roots per shoot. MS medium containing 1 mg L -1 BA+0.5 mg L-1 IAA + 100 mg L-1 Vitamin B2+100 mg L-1 citric acid is best for shoot proliferation and 1/2 strength MS medium with IBA 3 mg L-1 is best for root induction. © 2008 Asian Network for Scientific Information.