title: Artemisinin biosynthesis enhancement in transgenic Artemisia annua plants by downregulation of the β-caryophyllene synthase gene.
abstract: Artemisinin is an effective antimalarial drug isolated from the medicinal plant Artemisia annua L. Due to its increasing market demand and the low yield in A. annua, there is a great interest in increasing its production. In this paper, in an attempt to increase artemisinin content of A. ANNUA by suppressing the expression of β-caryophyllene synthase, a sesquiterpene synthase competing as a precursor of artemisinin, the antisense fragment (750 bp) of β-caryophyllene synthase cDNA was inserted into the plant expression vector pBI121 and introduced into A. annua by Agrobacterium-mediated transformation. PCR and Southern hybridization confirmed the stable integration of multiple copies of the transgene in 5 different transgenic lines of A. annua. Reverse transcription PCR showed that the expression of endogenous CPS in the transgenic lines was significantly lower than that in the wild-type control A. annua plants, and β-caryophyllene content decreased sharply in the transgenic lines in comparison to the control. The artemisinin content of one of the transgenic lines showed an increase of 54.9 % compared with the wild-type control. The present study demonstrated that the inhibition pathway in the precursor competition for artemisinin biosynthesis by anti-sense technology is an effective means of increasing the artemisinin content of A. annua plants.
authors.affiliation: Graduate University of the Chinese Academy of Sciences, Beijing, China.
authors.forename: Jian-Lin
authors.initials: JL
authors.lastname: Chen
authors.forename: Hua-Ming
authors.initials: HM
authors.lastname: Fang
authors.forename: Yun-Peng
authors.initials: YP
authors.lastname: Ji
authors.forename: Gao-Bin
authors.initials: GB
authors.lastname: Pu
authors.forename: Yan-Wu
authors.initials: YW
authors.lastname: Guo
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Huang
authors.forename: Zhi-Gao
authors.initials: ZG
authors.lastname: Du
authors.forename: Ben-Ye
authors.initials: BY
authors.lastname: Liu
authors.forename: He-Chun
authors.initials: HC
authors.lastname: Ye
authors.forename: Guo-Feng
authors.initials: GF
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism; D031172:Artemisia annua / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D037621:Artemisinins / Q000378:metabolism*; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D015536:Down-Regulation / Q000235:genetics; D004365:Drugs, Chinese Herbal / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008025:Ligases / Q000235:genetics*; D008516:Medicine, Chinese Traditional; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D010946:Plants, Medicinal; D010957:Plasmids; D000081223:Polycyclic Sesquiterpenes; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D036226:Seedlings / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D012717:Sesquiterpenes / Q000378:metabolism
chemical_list: D000890:Anti-Infective Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D037621:Artemisinins; D016373:DNA, Antisense; D018076:DNA, Complementary; D018744:DNA, Plant; D004365:Drugs, Chinese Herbal; D010940:Plant Proteins; D000081223:Polycyclic Sesquiterpenes; D012333:RNA, Messenger; D018749:RNA, Plant; D012717:Sesquiterpenes; C031327:artemisinin; C024714:caryophyllene; D008025:Ligases