Abstract

OPTIMIZATION OF AGROBACTERIUM TUMEFACIENS-MEDIATED GENETIC TRANSFORMATION FOR SYNTHESIS OF ANTI MALARIAL DRUG ARTEMISININ FROM PLANT ARTEMISIA ANNUA

*Kshitij Bhardwaj (M-Tech), A.K Mathur (PhD), Archana Mathur (PhD) Narsingh Verma, Shipra Bhardwaj (PhD), Neha Shukla (Msc)

Abstract

In this work, an attempt has been made to explore the factors influencing A. tumefaciens-mediated transformation of A. annua different combinations of hormone concentration, bacterial strain selection and their different ODs to optimize efficient transformation system. Finally, a system of high efficiency of genetic transformation and regeneration of A. annua can be developed .Different Agrobacterium strains used to transform Artemisia annua and their respective antibiotics for selection. EHA 105 and LBA 4404 were prepared for the elimination of Agrobacterium in plant in following concentration LBA 4404 - 500mg/l cefotoximin. EHA 105- 500mg/l carbenicilin.Stock was prepared as 1g/20ml and filter-sterilized before storing at -20ºC. From the results obtained strain EHA-105 appears to be superior to strain LBA 4404. The chromosome in Agrobacterium and activating potency of genes in virulence region are important internal factors influencing the infecting ability of agrobacteriam strains. Transformation efficiency can be improved by adding hormone to co-cultivation medium, because it can accelerate cell division of explants and maintain cell activity, consequently in favor of cell growth after transformation. However, in the present study, no major change in the regeneration frequency was observerved so far after the addition of hormone to co-cultivation medium while the rate was only 10% more in that case. The possible reason may be that shoot induction of A. annua happened at early stage.

Keywords: Agrobacteriam,, Artemisia annua, biotransformation, malaria.