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Overexpression of artemisinic aldehyde Δ11 (13) reductase gene–enhanced artemisinin and its relative metabolite biosynthesis in transgenic A rtemisia annua L.
Author(s) -
Yuan Yuan,
Liu Wanhong,
Zhang Qiaozhuo,
Xiang Lien,
Liu Xiaoqiang,
Chen Min,
Lin Zhi,
Wang Qiang,
Liao Zhihua
Publication year - 2014
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1234
Subject(s) - artemisinin , biosynthesis , metabolite , reductase , artemisia annua , chemistry , biochemistry , aldehyde , transgene , gene , aldehyde reductase , secondary metabolite , stereochemistry , biology , enzyme , catalysis , plasmodium falciparum , malaria , immunology
Artemisinic aldehyde Δ11 (13) reductase ( DBR 2) is the checkpoint enzyme catalyzing artemisinic aldehyde to form dihydroartemisinic aldehyde directly involved in artemisinin biosynthetic pathway. In the present study, DBR 2 was employed to engineer the biosynthetic pathway of artemisinin in transgenic plants of A rtemisia annua L . Seven independent transgenic plants of A . annua with DBR 2 overexpression driven by the cauliflower mosaic virus 35S promoter were obtained by A grobacterium ‐mediated genetic transformation and confirmed by genomic PCR . The results of real‐time q PCR analysis showed that the expression levels of DBR 2 gene in all the seven transgenic lines were significantly higher than in nontransgenic control. The high‐performance liquid chromatography analysis of artemisinin and its relative metabolites demonstrated that the contents of artemisinin and its direct precursor dihydroartemisinic acid were remarkably increased in the transgenic plants of A . annua with DBR 2 overexpression. Interestingly, it was also found that the contents of arteannuin B and its direct precursor artemisinic acid in the branch pathway competing against artemisinin biosynthesis were also improved in DBR 2‐overexpressed A . annua plants. The transgenic results in the present study indicated that DBR 2 is a useful structural gene in engineering the artemisinin biosynthetic pathway to develop genetically modified A . annua with the higher yield of artemisinin.