Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation

Abstract

An Agrobacterium tumefaciens-mediated transformation system was developed for Artemisia annua L. Using this system a cDNA encoding farnesyl diphosphate synthase (FDS placed under a CaMV 35S promoter) was transferred into A. annua via A. tumefaciens strain LB4404. Leaf or leaf discs were used as explants to be infected with A. tumefaciens and an optimal concentration of 20 mg/l kanamycin was applied to select kanamycin resistant shoots. Forty-five lines of resistance kanamycin shoots transformed with FDS were established. Analysis of PCR showed that at least 20 shoots transformed with the FDS gene were PCR positive. Southern blot analysis suggested the foreign FDS gene had been integrated into the A. annua genome, and Northern blot analysis revealed that the foreign FDS gene expressed at the transcriptional level in five shoot lines (F-1, F-4, F-61, F-62 and F-73 shoot lines). Analysis of artemisinin demonstrated that about 8 approximately 10 mg/g DW of artemisinin were then detected in transgenic plants regenerated from five shoot lines, this is about 2-3 times higher than that in the control.