In Silico Analysis and Feeding Assays of Some Genes in the Early Steps of Terpenoid Biosynthetic Pathway in Camellia Sinensis  

作　　者：Wei Xiang Yan Xu Fu-Min Wang Li-Ping Gao Ming-Jun Gao Zheng-Zhu Zhang Xiao-Chun Wan Shu Wei 

机构地区：[1]Key Laboratory of Tea Biochemistry and Biotechnology,Anhui Agricultural University [2]College of Life Sciences,Anhui Agricultural University [3]Agriculture and Agri-Food Canada,Saskatoon Research Center

出　　处：《茶叶》2013年第4期191-198,共8页Journal of Tea

基　　金：funded by the National Science Foundation in China (#31070614 to S.Wei);Fund for the Doctoral Program of Higher Education of The ministry of Education(#20123418110002,to S.Wei);the Program for Changjiang Scholars and Innovative Research Team in Universities (IRT1101 to Z.Z.Zhang);the "Twelfth Five-Year" National Key Basic Research and Development Project (973) in China (2012CB722903 to Xiao-Chun Wan)

摘　　要：Terpenoids play important roles in productivity and product quality in tea(Camellia sinensis),not only because volatile terpenoids contribute significantly to the aroma of teas,but terpenoid-derived light harvesting pigments and phytohormones such as strigolactones implement photosynthesis and branching in plants.All terpenoids are derived from isopentenyl diphosphate(IPP),dimethylallyl diphosphate(DMAPP) produced by methyl-erythritol-phosphate(MEP) and mevalonate(MVA) pathways,the early steps of isoprenoid biosynthesis.For the purpose to determine key genes in the MEP and MVA pathways in C.sinensis,five genes with complete coding sequence and three with partial sequence were obtained based on previously attained transcriptome data and polymerase chain reaction approach in this study.In silico analyses suggested that the five genes with full coding sequence individually encoded acetyl-CoA C-acetyltransferase(CsAACT),3-hydroxy-3-methylglutaryl-CoA synthase(CsHMGS),4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase(CsCMK),1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase(CsHDS),and farnesyl pyrophosphate synthase(CsFPS).Additionally,studies on metabolite-mediated gene expression were performed using tea cell suspensions and leaf-disc cultures.Our data showed that the addition of acetyl-CoA increased the transcript level of CsAACT and CsHMGS by 1.5- and 3-fold,respectively.The addition of deoxy-D-xylulose 5-phosphate(DXP) induced the expression of CsCMK and CsHDS by 2.3- and 1.5-fold,respectively.The CsFPS expression was approximately 2-fold higher when treated with DXP,IPP or DMAPP than the untreated control,but this induction was not observed with the acetyl-CoA treatment.Our studies indicated that expression of CsAACT,CsHMGS,CsCMK,CsHDS,and CsFPS were metabolite-mediated in a different extent.Terpenoids play important roles in productivity and product quality in tea （Camellia sinensis）,not only because volatile terpenoids contribute significantly to the aroma of teas,but terpenoid-derived light harvesting pigments and phytohormones such as strigolactones implement photosynthesis and branching in plants.All terpenoids are derived from isopentenyl diphosphate （IPP）,dimethylallyl diphosphate （DMAPP） produced by methyl-erythritol-phosphate （MEP） and mevalonate （MVA） pathways,the early steps of isoprenoid biosynthesis.For the purpose to determine key genes in the MEP and MVA pathways in C.sinensis,five genes with complete coding sequence and three with partial sequence were obtained based on previously attained transcriptome data and polymerase chain reaction approach in this study.In silico analyses suggested that the five genes with full coding sequence individually encoded acetyl-CoA C-acetyltransferase （CsAACT）,3-hydroxy-3-methylglutaryl-CoA synthase （CsHMGS）,4-（cytidine 5 ＇-diphospho）-2-C-methyl-D-erythritol kinase （CsCMK）,1-hydroxy-2-methyl-2-（E）-butenyl-4-diphosphate synthase （CsHDS）,and famesyl pyrophosphate synthase （CsFPS）.Additionally,studies on metabolite-mediated gene expression were performed using tea cell suspensions and leaf-disc cultures.Our data showed that the addition of acetyl-CoA increased the transcript level of CsAACT and CsHMGS by 1.5-and 3-fold,respectively.The addition of deoxy-D-xylulose 5-phosphate （DXP） induced the expression of CsCMK and CsHDS by 2.3-and 1.5-fold,respectively.The CsFPS expression was approximately 2-fold higher when treated with DXP,IPP or DMAPP than the untreated control,but this induction was not observed with the acetyl-CoA treatment.Our studies indicated that expression of CsAACT,CsHMGS,CsCMK,CsHDS,and CsFPS were metabolite-mediated in a different extent.

关 键 词：萜类化合物 生物合成途径 基因表达 茶树 早期 硅片 法呢基焦磷酸合酶 聚合酶链反应 

分 类 号：TQ914.1[化学工程]