Transformation of Amorphadiene Synthase and Antisilencing P19 Genes into L. and its Effect on Antimalarial Artemisinin Production.

The low content of artemisinin related to the biosynthetic pathway is influenced by the role of certain enzymes in the formation of artemisinin. The regulation of genes involved in artemisinin biosynthesis through genetic engineering is a choice to enhance the content. This research aims to transform and gene as an antisilencing into and to see their effects on artemisinin production. The presence of and genes was confirmed through polymerase chain reaction (PCR) products and sequencing analysis. The plasmids, which contain and/or genes, were transformed into , and then inserted into leaves and hairy roots of A. annua by vacuum and syringe infiltration methods. The successful transformation was checked through the GUS histochemical test and the PCR analysis. Artemisinin levels were measured using HPLC. The percentages of the blue area on leaves by using vacuum and syringe infiltration method and on hairy roots were up to 98, 92.55%, and 99.00% respectively. The - sample contained a higher level of artemisinin (0.18%) compared to other samples. Transformed hairy root with co-transformation of - contained 0.095% artemisinin, where no artemisinin was found in the control hairy root. The transformation of and genes into A. annua plant has been successfully done and could enhance the artemisinin content on the transformed leaves with - up to 2.57 folds compared to the untransformed leaves, while for , cotransformed and were up to 2.25, 1.29, and 1.14 folds respectively. Antisilencing gene could enhance the transformation efficiency of and artemisinin level in A. annua.