Abstract

A method for mass propagation of Tylophora indica (Burm. f.) Merill. (Asclepiadaceae) from leaf explants and extraction of tylophorine was developed. Explants cultured on 9.0 µM α-naphthalene acetic acid (NAA) and 4.65 µM kinetin (K) resulted in callus formation under dark conditions after 7 to 8 days of culturing, whereas formation of nodular meristemoids was observed on 8.8 µM 6-benzyladenine (BA). Eventually, both the cultures developed green leafy shoots on their transfer to 9.84 µM 6-benzyladenine shoot inducing medium. Microshoots thus formed were cultured on basal MS root inducing medium, which resulted in the formation of long healthy roots within 10 to 12 days. Plantlets were successfully hardened and transferred to field conditions. Tylophorine was extracted from the leaves of regenerated plants using organic solvents such as like hexane, chloroform and dichloromethane and separated on high performance thin layer chromatography (HPTLC) using toluene: chloroform: ethanol: ammonia (4:3.5:1.5) as mobile phase. Amount of tylophorine obtained was 80 and 71 µg/ml from callus raised and directly cultured in vitroplants respectively.

Key words: Tylophora indica, high performance thin layer chromatography, tylophorine, 6-benzyladenine, α-naphthalene acetic acid.