Genetic transformation in Artemesia annua L. for hairy root induction and enhancement of secondary metabolites

V.M. Shaneeja, R. Keshavachandran, Priyanka James, P.A. Nazeem

Abstract


Hairy root induction was attempted in Artemesia annua for enhancing the production of the secondary metabolite artemisinin, a powerful antimalarial drug. Leaf segments were regenerated on MS medium with 0.5 mg l-1 BAP alone or on 0.1 mg l-1 NAA and 2 mg l-1 BAP. Agrobacterium rhizogenes strain MTCC 2364 showed very fast growth on YEB, YEP, NA, Xanthomonas and LBA media and since the growth was poor on YEM media, this medium was selected for growing the bacteria to obtain single cell colonies. For the Agrobacterium strains ATCC 15834 and A4, both NA and YEB were good. Confirmation of transformation was done through opine analysis using high-voltage paper electrophoresis and PCR assay. Southern hybridization was also carried out further for confirming the presence of rol B gene of TL-DNA in the transformed roots. ATCC 15834 was the most efficient Agrobacterium strain to induce the hairy roots in Artemisia. When estimated using TLC, there was increase in artemisinin content of ATCC 15834 induced hairy roots grown in media elicited with the homogenate (from 0.0039 per cent FW to 0.0042 per cent FW).


Keywords


Agrobacterium; Artemesia; Artemisinin; Genetic modification; Hairy root; Secondary metabolite; Transformation

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A KAU publication [CODEN: JTAGEI; ISSN 0971-636X; eISSN 0973-5399]