In Vitro Plantlet Regeneration from Alysicarpus monilifer, a Source of Hepatoprotective Drug

Abstract

A plantlet regeneration protocol has been developed for Alysicarpus monilifer, a medicinal plant that is a source of hepato-protective drugs. Callus was induced from mature cotyledonary leaves from 4 to 5 days old seedling on Murashige and Skoog (MS) medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxyacetic acid (2, 4-D). Proliferation of cultures occurred on MS medium with 1.0 mg l−1 each of 2,4-D, 6-benzylaminopurine (BA) and kinetin (Kin). Shoot regeneration from proliferated callus was influenced by a number of factors namely plant growth regulators (PGRs), gelling agents, culture vessels and carbohydrate source. The highest (85.6 %) shoot regeneration was recorded in 250 ml culture flasks on agar gelled MS medium + 0.1 mg l−1 α-naphthalene acetic acid (NAA) + 1.0 mg l−1 each of BA, Kin and 2-isopentenyladenine (2iP) + 1 % glucose and 2 % maltose in addition to the usual 3 % sucrose. The shoots differentiated on PGRs, free MS medium, were stronger and longer than the shoots developed on MS medium containing PGRs (0.1 mg l−1 NAA + 1.0 mg l−1 each of BA, Kin and 2iP) with different leaf morphology and were easy to root. Rooting of the regenerated shoots was achieved both in vitro and ex vitro. About 80.4 % of the shoots rooted in vitro on half strength MS medium containing 1.0 mg l−1 indole-3-butyric acid (IBA), while 84.9 % of the shoots rooted under the ex vitro condition when treated with 250 mg l−1 IBA for 5 min. The plants were hardened in the green house and showed 85 % survival rate.