High-frequency shoot regeneration from leaf explants through organogenesis in bitter melon (Momordica charantia L.)

Abstract

An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g l(-1) sucrose, 2.2 g l(-1) Gelrite, and 7.7 mu M naphthalene acetic acid (NAA) with 2.2 mu M thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 mu M TDZ, 2.2 mu M NAA, and 3.3 mu M silver nitrate (AgNO(3)). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 mu M gibberellic acid (GA(3)). The elongated shoots were rooted in MS medium supplemented with 4.0 mu M indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.