Efficient protocol for regeneration of transformed artemisia annua plants from hairy roots

Abstract

Elite genotypes from high yielding variety of Artemisia annua L.  (Jeevanraksha, CIMAP Lucknow) were regenerated form nodal explants and maintained in MS culture medium with BAP and NAA. The leaves of tissue culture grown plants were inoculated with Agrobacterium rhizogenes strain LBA9402 containing the selective marker gene NPT II. Hairy roots emerged from the leaves, 8-10 days after inoculation and were maintained in MSO liquid/ solid medium with kanamycin as the selective agent. Hairy root line was selected on the basis of high artemisinin content. Regeneration from hairy roots was obtained in half strength MS solid medium containing a combination of cytokinins and auxins (BAP, Kinetin, NAA and IBA). Transgenic plants showed typical transgenic morphology and profuse growth of plageotropic roots. Transformed hairy root cultures and transgenic plants can be used as an experimental system for studies related to induction of secondary metabolite synthesis.