Overexpression of the cytochrome P450 monooxygenase (cyp71av1) and cytochrome P450 reductase (cpr) genes increased artemisinin content in Artemisia annua (Asteraceae).
Finding  an efficient and affordable treatment against malaria is still a challenge for  medicine. Artemisinin is an effective anti-malarial drug isolated from Artemisia  annua. However, the artemisinin content of A. annua is very low. We  used transgenic technology to increase the artemisinin content of A. annua by  overexpressing cytochrome P450 monooxygenase (cyp71av1) and cytochrome  P450 reductase (cpr) genes. CYP71AV1 is a key enzyme in the artemisinin  biosynthesis pathway, while CPR is a redox partner for CYP71AV1. Eight  independent transgenic A. annua plants were obtained through Agrobacterium  tumefaciens-mediated transformation, which was confirmed by PCR and  Southern blot analyses. The real-time qPCR results showed that the gene cyp71av1 was highly expressed at the transcriptional level in the transgenic A.  annua plants. HPLC analysis showed that the artemisinin content was  increased in a number of the transgenic plants, in which both cyp71av1 and cpr were overexpressed. In one of the transgenic A. annua plants,  the artemisinin content was 38% higher than in the non-transgenic plants. We  conclude that overexpressing key enzymes of the biosynthesis pathway is an  effective means for increasing artemisinin content in plants.
Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
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D031172:Artemisia annua / Q000201:enzymology* / Q000235:genetics*; D037621:Artemisinins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000235:genetics; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012038:Regeneration / Q000235:genetics; D014170:Transformation, Genetic
D037621:Artemisinins; D003577:Cytochrome P-450 Enzyme System; C031327:artemisinin; D009251:NADPH-Ferrihemoprotein Reductase