Micropropagation ofMucuna pruriens(L.) DC and determination of tyrosinase

Abstract

The protocol for in vitro propagation of Mucuna pruriens (white seed) and the extraction of laccase-like enzyme –tyrosinase; has been standardized. Root, internode, axillary buds, seed and leaf were used as explants for callus induction. Maximum compact green calli were produced by using different concentrations of NAA and combinations of IBA+BAP. Maximum white friable calli were produced by using different concentrations of 2, 4-D and combination with Kinetin. Direct organogenesis and indirect organogenesis were derived from various explants of M. pruriens by using different combinations of auxins and cytokinins. The high phenolic content of M. pruriens caused initial browning which was avoided by applying 0.01% of acid washed charcoal into 1 liter MS medium. Instead of charcoal, ascorbic acid (50 mg/L), citric acid (25 mg/L) and L-arginine (25 mg/L) were added into 1 L MS medium and kept in dark condition to overcome browning effect. The laccase-like enzyme tyrosinase was determined by qualitative analysis and quantified.