The Transformation Of IaaM Into Artemisia Annua And Its Affection On The Glandular Trichome Development

Malaria is a severe infectious disease transmitted by parasite. The artemisinin is now considered to be the best drug in anti-malarial especially for cerebral malaria and chloroquine resistant falciparum malaria. Now the only source of artemisinin was extracted from Artemisia annua. The low content of artemisinin in the wormwood limited its production and efficiency. The biotechnology will be a good choice to increase the production of artemisinin.A transgenic protocol is designed to regulate the glandular trichome development in this study. The G12promoter and the E6promoter that are specifically expressed in trichome are isolated by PCR and recombined with the iaaM gene, a IAA synthesis gene originally from bacterium, respectively.The G12promoter cloned from Arabidopsis thaliana is recombined with iaaM gene and cloned into pWM101Ti plasmid. The vector is designed as pWM101-G12::iaaM. The E6promoter cloned from Gossypium is recombined with iaaM and cloned into pWM101that is previously cloned and be designed as pWM101-E6::iaaM. The two vectors are transformed separately into Agrobacterium tumefaciens GV3101for plant transformation.The Artemisia annua tissue culture system and transformation regeneration system ware optimized at first. The optimized hormone concentration and combination can make higher frequency of shoot and root induction. The cotyledons are considered to be the optimum explant after a comparative trial. The transgenic analysis of hygromycin resistance deduced that the8mg/L hygromycin was the suitable selective concentration for screening of the transforms.The GL2::iaaM and E6::iaaM were transformed into the Artemisia annua via cotyledon explant co-culture infected with Agrobacterium tumefaciens. Transformed plantlets of E6::iaaM were screened out by hygromycin selection. The resistance plantlets are subjected to PCR molecular detection. Some5 transformed plantlets are identified. But4of the plantlets were yellowing and withered that only one plantlet survived. The transgenic plantlet has an increased density of the glandular trichomes in its leave. The increased ratio is about21%. The G12::iaaMtransforms are still in screening and shoots development. Because of the limited number of the transformed plantlets that the field trial and statistical analysis can’t be conduct yet.