In vitro plant regeneration of Aristolochia indica through axillary shoot multiplication and organogenesis

Protocols for in vitro plant regeneration via axillary and adventitious shoot regeneration were established in an important medicinal plant, Aristolochia indica L. (Aristolochiaceae). Basal Murashige and Skoog's (MS) medium supplemented with 0.54 mu M alpha-naphthaleneacetic acid (NAA) and 13.31 mu M benzyladenine (BA) induced the maximum number of shoots (45-50) from shoot tip and nodal segment cultures. Phenolic accumulation in leaf and internodal stem derived callus cultured in MS medium containing NAA or 2,4-dichlorophenoxyacetic acid and BA or kinetin was controlled by the addition of 1.0 mg l(-1) phloroglucinol (PG) to the callus induction medium. Basal medium supplemented with 2.69 mu M NAA, 13.31 mu M BA and 1.0 mg l(-1) PG induced the best results in terms of shoot bud regeneration from leaf derived callus. Direct de novo development of shoots from leaf segments was achieved using 13.31 mu M BA along with 50 mg l(-1) activated charcoal. The microshoots were rooted in White's medium supplemented with 2.46 mu M indolebutyric acid. More than 85% of rooted plants survived in the soil.