Acorus calamus L. is a species of enormous medicinal and economic importance. In vitro propagation of this plant was achieved using axillary bud explant. In the present investigation, naturally grown axillary bud and rhizome explants were cultured on standard MS and B5 medium supplemented with different concentration and combination of cytokinines and auxines. The best shoot proliferation was observed in MS medium containing Kn (1.0mg/l) +IBA (0.5mg/l) with 3.33±0.58 nos. of Shoots, 7.33±0.58 nos. of roots and 15.33±0.58 nos. leaves. In B5 medium best results found in Kn (1.5mg/) + NAA (1.0mg/l) with 2.67±0.58 nos. shoots, 3.67±0.58 nos. of roots and11.67±0.58 nos. of leaves. They were then transplanted in soil: sand: cow dung mixture (1:1:2) and kept in shade for 4 to 5 weeks and then transferred to field for one month. Survival rate was found 80 % in MS medium and 100 % in B5 medium. The present investigation was carried out with a view to standardize an in vitro culture technique for mass propagation of this important medicinal plant species and was found successful.