Abstract: Callus induction from leaf explant of Achyranthes aspera L. had been achieved using Murashige and Skoog (MS) medium supplemented with various plant growth regulators under light and dark incubation conditions. Callusing potential on B5 medium had also been evaluated. Our results showed that Kinetin (KIN) was inefficient in callus formation. Leaf explant inoculated on MS+2,4-D (1 mg/L-1) or MS+2,4,5-T (7 and 8 mg/L-1) media and incubated under light produced good callus. But colour of the callus formed in 2,4-D medium became brown after 2 weeks. Better callusing response was seen in MS+2,4,5-T (8 mg/L-1) incubated under dark. Subculture of callus initiated using MS+2,4,5-T (8 mg/L-1) under light condition onto MS medium without any plant growth regulator, BM (basal medium) under dark incubation resulted in yellow green, fleshy and soft callus without browning. B5 medium was not good in callus formation under any incubation conditions.