牡丹PsDXR和PsMCS基因的克隆、表达分析及亚细胞定位  

Cloning,Expression Analysis and Subcellular Localization of PsDXR and PsMCS Genes in Tree Peony(Paeonia suffruticosa)

作　　者：李紫瑶 刘冰 胡增辉[1,2] 吴静 冷平生[1,2] LI Zi-Yao;LIU Bing;HU Zeng-Hui;WU Jing;LENG Ping-Sheng(College of Landscape Architecture,Beijing University of Agriculture,Beijing 102206;Beijing Laboratory of Urban and Rural Ecological Environment,Beijing University of Agriculture,Beijing 102206)

机构地区：[1]北京农学院园林学院,北京102206 [2]北京农学院,城乡生态环境北京实验室,北京102206

出　　处：《农业生物技术学报》2023年第4期730-740,共11页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划(2018YFD1000406);国家自然科学基金(31800602);中央高校基本科研业务费专项资金(BFUKF202211)。

摘　　要：萜烯类化合物是牡丹(Paeonia suffruticosa)的主要花香成分,1-脱氧木酮糖-5-磷酸还原酶(1-deoxyxylulose-5-phosphate reductoisomerase,DXR)和2-甲基赤藓糖-2,4-环二磷酸合成酶(2-Cmethylerythritol-2,4-cyclodiphosphate synthase,MCS)作为2-C-甲基-D-赤藓糖醇-4-磷酸(2-C-methyl-Derythritol 4-phosphate,MEP)途径中的关键酶,在植物萜烯合成中起着重要作用。为探究PsDXR和PsMCS基因在牡丹萜烯合成途径中的作用,本研究以牡丹’皇冠’cDNA为模板,克隆了PsDXR(GenBank No.ON092613)和PsMCS基因(GenBank No.ON092614),对其进行了生物信息学分析和亚细胞定位,并通过qRT-PCR分析了其在牡丹’皇冠’不同花发育时期和组织中的表达。结果显示,PsDXR和PsMCS基因cDNA编码全长分别为1401和702 bp,分别编码466和233个氨基酸。2个基因编码的蛋白均为稳定亲水性蛋白,PsDXR蛋白具有DXP_reductoisom和DXP_redisom_C保守结构域,PsMCS蛋白具有MECDP特有结构域。亚细胞定位结果显示2个蛋白均定位于叶绿体。PsDXR和PsMCS基因在花的不同发育时期的表达模式均为先升高后降低;在不同组织中,2个基因均具有显著的表达差异性(P<0.05)。研究结果表明PsDXR和PsMCS基因在牡丹MEP代谢途径中可能起着重要的作用。本研究为牡丹在萜烯类物质的生物合成中提供分子研究依据。Terpenoids are the main floral components of tree peony(Paeonia suffruticosa).1-deoxyxylulose-5-phosphate reductoisomerase(DXR)and 2-C-methylerythritol-2,4-cyclodiphosphate synthase(MCS)play important roles as key enzymes in the 2-C-methyl-D-erythritol-4-phosphate(MEP)pathway in plant terpene synthesis.To investigate the role of PsDXR and PsMCS genes in the terpene synthesis pathway of tree peony,the PsDXR(GenBank No.ON092613)and PsMCS genes(GenBank No.ON092614)were cloned using the’Huang Guan’cDNA of tree peony in this study,and their bioinformatics analysis,subcellular localization and qRT-PCR were performed to analyze the expression in different flower developmental stages and tissues of tree peony’Huang Guan’.The results showed that the c DNAs of PsDXR and PsMCS genes were 1401 and 702bp in length,encoding 466 and 233 amino acids,respectively.The proteins encoded by the 2 genes were stable hydrophilic proteins.The PsDXR protein had the conserved domains of DXP_reductoisom and DXP_redisom_C,and the PsMCS protein had the MECDP-specific domain.The results of subcellular localization showed that both proteins were localized in the chloroplast.The expression patterns of PsDXR and PsMCS genes both increased and then decreased in flower development stages;both 2 genes were differentially expressed in different tissues(P<0.05).The results indicated that PsDXR and PsMCS genes might play important roles in the MEP metabolic pathway of tree peony,provides a molecular basis for the biosynthesis of terpenes in tree peony.

关 键 词：牡丹 牡丹1-脱氧木酮糖-5-磷酸还原酶(PsDXR)基因 牡丹2-甲基赤藓糖-2 4-环二磷酸合成酶(PsMCS)基因 基因克隆 表达分析 亚细胞定位 

分 类 号：S685.11[农业科学—观赏园艺]