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Artemisinin is an effective antimalarial sesquiterpene lactone synthesized in  Artemisia annua . Various transcription factors have been previously reported that can influence the biosynthesis of artemisinin; however, the effect of YABBY family transcription factors on artemisinin biosynthesis was unknown. In the present study, we cloned and characterized AaYABBY5: a homolog of MsYABBY5 in  Mentha spicata  which is involved in modulating the monoterpenes, as a positive regulator of artemisinin biosynthesis in  A. annua . AaYABBY5 was found localized to the nucleus, and its expression was found to be induced by exogenous methyl jasmonic acid (MeJA) treatment. In the dual-luciferase reporter assay, it was found that AaYABBY5 significantly increased the activities of promoters of amorpha-4,11-diene synthase ( ADS ), cytochrome P450 monooxygenase ( CYP71AV1 ), double-bond reductase 2 ( DBR2 ), and aldehyde dehydrogenase 1 ( ALDH1 ) genes. Yeast one hybrid assay showed that AaYABBY5 directly bonds to the promoters of  CYP71AV1  and  DBR2  genes. Quantitative real-time polymerase chain reaction (qPCR) of  AaYABBY5  overexpression and  AaYABBY5  antisense plants revealed a significant increase in the expression of  ADS ,  CYP71AV1 ,  DBR2 , and  ALDH1  in  AaYABBY5  overexpression plants and a significant decrease in the expression of these genes in  AaYABBY5  antisense  A. annua , respectively. Furthermore, the results of high-performance liquid chromatography (HPLC) showed that the artemisinin and its precursor dihydroartemisinic acid were significantly increased in the  AaYABBY5  overexpression plants while  AaYABBY5  downregulation resulted in a significant decrease in the concentration of artemisinin. Taken together, these results explicitly represent that AaYABBY5 is a positive regulator of artemisinin biosynthesis in  A. annua .

frontiers in plant science

The YABBY Family Transcription Factor AaYABBY5 Directly Targets Cytochrome P450 Monooxygenase (CYP71AV1) and Double-Bond Reductase 2 (DBR2) Involved in Artemisinin Biosynthesis in Artemisia Annua