Micropropagation and Agrobacterium rhizogenes mediated transformation studies in Mucuna pruriens (L.) DC.

Abstract

An efficient protocol for micropropagation and Agrobacterium rhizogenes mediated transformation has been standardized for Mucuna pruriens (L.) DC. For micropropagation, shoot tips from aseptically germinated seedlings were inoculated on MS medium supplemented with different concentrations (2–20 µM) of phytohormones. Of the various phytohormones tried, kinetin (14 µM) resulted in 4 shoots/explant after four weeks of culture. Rooting was best obtained on half-strength MS medium supplemented with 6 µM concentration of NAA. Regenerated plants were hardened with 75% of survival frequency. For hairy root induction,different explants were infected by two strains of Agrobacterium rhizogenes (MTCC 2364 and MTCC 532) and effect of manual wounding and acetosyringone was studied. Of the two strains tried, MTCC 2364 showed hairy roots induction in leaf explant only by manual wounding method. While in case of acetosyringone (50-250 µM), mediated method hairy root induction was observed in all the explants studied except leaf. Out of the two strains used, MTCC 2364 showed better results in terms of roots emerged/explant and number of days required for root induction as compared to the strain MTCC 532.