Effect of various carbon and nitrogen sources, PGRs and their precursors, on the callus growth and L-dopa production by the stem and leaf derived callus of Mucuna pruriens were studied. Five different carbon sources were glucose, fructose, sucrose, mannose and raffinose. Glucose and sucrose were almost equally effective in both promoting the growth as well as L-dopa content. The four nitrogen sources were phenylalanine, tyrosine, glutathione and casein hydrolysate. Tyrosine was found to be most effective nitrogen source at 25 mgl-1. Leaf derived callus showed more increase in L-dopa than stem. Phenylalanine (10 mgl-1) along with tyrosine (25 mgl-1) could raise the L-dopa content (i.e. % of dry weight) from 1.4% of control to 1.7%. Casein hydrolysate was not very effective in L-dopa production. The PGRs tried were BA, Kin, Ad.SO4; IAA, its precursor shikimic acid and an intermediary compound tryptophan; and gibberellic acid and its precursor mevalonic acid. 1.0 mgl-1 IAA was most effective in promoting both growth and L-dopa content, whereas gibberellic acid and mevalonic acid inhibited both L-dopa content as well as growth. Incorporation of best nitrogen source (25 mgl-1 tyrosine) + most effective PGR (1 mgl-1IAA) to maintenance medium enhanced the L-dopa production to 1.95%.