茶树甲羟戊酸激酶基因CsMVK克隆及表达特性分析

发布时间：2019-01-15 06:48

【摘要】：【目的】克隆茶树甲羟戊酸激酶基因(Cs MVK),分析其生物学信息及在不同组织及乌龙茶做青过程中的表达特性,为研究茶叶加工过程中香气形成机制提供参考。【方法】RT-PCR扩增Cs MVK基因,应用生物信息学软件对其编码蛋白(Cs MVK)的氨基酸序列进行预测分析,通过实时荧光定量PCR(q PCR)分析Cs MVK在茶树不同组织及乌龙茶做青过程中的表达特性。【结果】克隆获得的Cs MVK基因(Gen Bank登录号MF668187)全长1455 bp,开放阅读框(ORF)长度为1164 bp,编码387个氨基酸,蛋白质分子量41.18 k D,理论等电点(p I)5.88。Cs MVK蛋白具有GHMP_kinases_N domain和GHMP_kinases_C domain 2个功能结构域,定位于细胞质中,不存在跨膜结构和信号肽。系统发育进化树分析结果显示,Cs MVK与三七、常春藤的MVK聚为一类,表明茶树与三七、常春藤的亲缘关系最近。q PCR检测结果显示,Cs MVK基因在果实中的表达量显著高于其他组织(P0.05,下同),在不同组织中的表达量排序为:果实根叶茎花;在乌龙茶做青过程中,从鲜叶到晒青叶,Cs MVK基因的表达量上升,晒青到一摇过程中表达量下降,经过3次摇青,其表达量持续升高并在杀青前达最大值,与其他做青阶段差异显著。【结论】Cs MVK基因表达与茶叶香气品质形成有关。
[Abstract]:[objective] to clone the methylvalerate kinase gene (Cs MVK),) from tea plants and analyze its biological information and expression characteristics in different tissues and Oolong tea. [methods] the Cs MVK gene was amplified by RT-PCR and the amino acid sequence of its encoded protein (Cs MVK) was predicted by bioinformatics software. The expression characteristics of Cs MVK in different tissues of tea plant and oolong tea were analyzed by real-time fluorescence quantitative PCR (q PCR). [results] the total length of Cs MVK gene (Gen Bank accession number (MF668187) was 1455 bp,. Open reading frame (ORF) encodes 387 amino acids with a molecular weight of 41.18kD and a length of 1164 bp,. The theoretical isoelectric point (p I) 5.88.Cs MVK protein has two functional domains, GHMP_kinases_N domain and GHMP_kinases_C domain. Located in the cytoplasm, there was no transmembrane structure or signal peptide. Phylogenetic tree analysis showed that the MVK of, Cs MVK and Panax notoginseng and Ivy were clustered together, indicating that the phylogenetic relationship between tea and Panax notoginseng and ivy was recently detected by. Q PCR. The expression of Cs MVK gene in fruit was significantly higher than that in other tissues (P0.05, the same below), and the order of expression in different tissues was as follows: fruit, root, leaf, stem and flower; In the process of oolong tea green, the expression of, Cs MVK gene increased from fresh leaf to sun-green leaf, and decreased during sun-green to first shake. After three times of shaking green, the expression of, Cs MVK gene increased continuously and reached the maximum before killing green. [conclusion] the expression of Cs MVK gene is related to the formation of tea aroma quality.
【作者单位】：福建农林大学园艺学院/茶学福建省高校重点实验室;
【基金】：福建省“2011协同创新中心”中国乌龙茶产业协同创新中心专项项目(闽教科[2015]75号) 国家现代农业(茶叶)产业技术体系建设专项资金项目(CARS-19) 福建农林大学科技创新专项基金项目(CXZX2016117,CXZX2017181)
【分类号】：S571.1

【相似文献】