In vitro plant regeneration through somatic embryogenesis in Anaphyllum wightii Schott

Anaphyllum wightii Schott. is an ethnomedicinally significant plant endemic to the southern region of Western Ghats. The present study aimed to develop an efficient protocol for the in vitro propagation of the plant through somatic embryogenesis. Fresh petioles were selected as explants for the experiment. The medium used was half-strength Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthaleneacetic acid (NAA). Both 2,4-D and NAA showed embryogenic callus induction from petiole explants after up to 4 wk of culture. NAA at 2 mg L-1 showed the highest percentage of embryogenic callus induction (83.33 +/- 8.80%). Histological and stereomicroscopic observations of the embryogenic callus revealed various stages of somatic embryos indicating an asynchronous type of embryogenesis. Full-strength MS medium containing 3 mg L-1 6-benzylaminopurine induced the maximum number of shoots per callus (6.00 +/- 0.58) after 30 d of culture for NAA-induced calluses. The highest rooting response was obtained with half-strength MS medium fortified with 0.5 mg L-1 indole-3-butyric acid. The in vitro rooted plantlets were hardened by transferring to small plastic pots containing sand and garden soil (1:1) and showed a 76% survival rate after 4 wk. Thus, the present work developed an efficient in vitro protocol for the conservation of this plant species and also contributes to the study of the embryonic development.