Abstract
Abstract An easy, efficient and highly reproducible regeneration system was established through organogenesis from leaf base mediated callus of an ethnobotanical hairy shrub Abutilon indicum L. sweet, which is documented to possess pharmaceutically important phytochemicals. Among various explants, leaf bases produced significant callus induction (89%) with 140 mg fresh weight on Murashige and Skoog (MS) medium supplemented with 11.31 μM 2, 4-Dichlorophenoxyacetic acid (2, 4-D). Combinations of 2, 4-D and kinetin (KIN) increased the biomass (191 mg) of embryogenic calli. Regeneration medium with a combination of 8.88 μM of 6-amino benzyl purine (BAP) and 8.06 μM of naphthalene acetic acid in MS basal media significantly influenced the initiation of shoot primordium with 91% of regeneration. Proficiently regenerated shoot apical meristems (SAMs) produced higher frequency of multiple shoot induction on 4.10 μM of zeatin. The fully regenerated mature shootlets produced high root biomass (791.67 ± 48.05 mg) on half - strength MS with 4.54 μM indole-3-butyric acid (IBA) with an excellent mat root system for efficient acclimatization. In vitro established plantlets were successfully acclimatized under ex vitro conditions with high frequency. Presence of homologous DNA banding pattern with Single Primer Amplification Reaction markers confirmed the genetic similarity. FTIR and GC-MS spectrum showed similar metabolic profiles between wild and in vitro regenerated plantlets. Thus the protocol is reliable for in vitro regeneration through leaf base derived callus which could pave a way to the large scale production of secondary metabolite through biotechnological approaches.
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