An efficient transformation system for hairy-root induction in Achyranthes aspera L. was established and investigated as a novel method for the production of 20-hydroxyecdysone. Four Agrobacterium rhizogenes strains (MTCC 2364, NCIM 5140, A4, and ATCC 15834) were examined for their frequency of transformation in the presence of various concentrations of acetosyringone. Strain ATCC 15834 showed significantly higher transformation efficiency and was capable of inducing hairy roots from different explants (young leaf, hypocotyls, cotyledon, and stem segments) of A. aspera in the presence of 100 µM acetosyringone. The hairy root transformation of the lines (AS001 and AS002) was confirmed by PCR amplification of rol B locus using primers specific for rol B gene. Hairy roots grown in the Murashige and Skoog (MS) liquid medium augmented with 30 g/L sucrose exhibited the highest biomass accumulation and this medium was found to be superior to Gamborg’s B5 medium and Whites root culture medium. HPLC and LC-Q-TOF analyses of the hairy-root extract of A. aspera revealed the ability of hairy-root lines to synthesise the phytoecdysteroid 20-hydroxyecdysone.
Acta Physiologiae Plantarum – Springer Journals
Published: Oct 22, 2017
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