The present study focused on the establishment of micropropagation protocol for the high valuePluchea (P.) indica (L.) Less., genotype, an important medicinal plant and evaluation of the diureticactivity of the leaf extract of the tissue cultured plant. Leaf explants, nodal segments and shoottips were cultured in MS medium supplemented with auxin and cytokinin and theircombinations. With the objective of inducing callus giving rise to new adult plants, naphthaleneacetic acid was found to be most effective for (80%) for callus induction. The methanolic extract ofleaves of the micropropagated P. indica was investigated for its diuretic activity in Wistar albinorats. Urinary excretion parameters were studied for evaluation of diuretic activity usingFrusemide (20 mg/kg, p.o.) as standard. The extract showed significant diuretic activity at thedoses of 100, 200 and 300 mg/kg. p.o. An oral acute toxicity study for the extract was carried outand the LD50 value was found to be 2,825 mg/kg body weight.