PRODUCTION OF ARTEMISININ AND ITS DERIVATIVES IN HAIRY ROOTS OF ARTEMISIA DUBIA INDUCED BY rolA GENE TRANSFORMATION

摘要

Artemisinin and its derivatives are phytochemical constituents of genus Artemisia. Demand of these plant secondarymetabolitesis increasing due to their immense therapeutic significance. Besides their established antimalarial role, recentstudies have also disclosed their anticancer potentials. It has made imperative to develop new and efficient sources of thesecompounds. Inherent synthetic challenges give biological sources preference over chemical synthesis of artemisinin and itsderivatives. Therefore, genetic improvement of plants and, rather less preferentially, microbes is focus of current research togain increase productivity of these valuable drugs. This study has analyzed A. dubiaas potential source of artemisinin and itsderivatives. Transformation of Artemisia dubia was carried out using A. tumefaciens strain LBA 4404 containing rolA geneconstructed on pRB 29. Healthy and acclimatizable transgenic plants were produced using optimized concentrations of BAPand NAA. Previously acclimatized rol ABC transgenic plants were also In vitro regenerated for comparative analysis ofartemisinin and its derivatives. PCR amplification of rolA gene was done to confirm the integration of T-DNA in transgenicplants.TLC analysis was performed to evaluate comparative production of artemisinin and derivatives in rolA and rol ABCtransgenic A. dubia. It revealed that rolA transgenic plants contain comparable amounts of these metabolites. Both typeoftransgenic plants manifested the enhancement of other uncharacterized compounds as well. Besides systematicoptimization of In vitro regenerative protocol for Artemisia dubia, relative regeneration ability of rol transgenic and controlplants was also assessed at four regenerative stages. It was observed that unlike control, rol transgenic plants showed bestroot induction only on combination of auxins and cytokines. It was concluded that rol genes transformation of plants is anefficient tool to enhance their secondary metabolites production. RolA transgenic A. dubiaare cultivable, viable and efficientsource of artemisinin and its derivatives to meet their ever-growing demands.