茶树甲羟戊酸焦磷酸脱羧酶基因CsMVD的克隆与表达分析
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国家自然科学基金(31270735);


Cloning and Expression of Mevalonate Diphosphate Decarboxylase Gene CsMVD in Tea Plant (Camellia sinensis
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    摘要:

    该研究在茶树转录组测序的基础上,以茶树品种‘福鼎大白茶’的芽叶为供试材料,采用RTPCR技术,克隆茶树萜类化合物合成途径中的限速酶——甲羟戊酸焦磷酸脱羧酶全长cDNA序列,命名为CsMVD(GenBank登录号为MF772780);该基因全长1 585 bp,其ORF为1 266 bp,编码421个氨基酸,蛋白质分子量46.45 kD,理论等电点7.10。CsMVD蛋白可能定位于细胞质中,具有MVD1 superfamily的保守结构域,不存在跨膜结构和信号肽,具有多个磷酸化位点,部分保守的氨基酸残基决定了蛋白的催化反应特异性和活性。系统进化分析表明,CsMVD与新疆紫草(Arnebia euchroma)MVD的亲缘关系最近。荧光定量PCR结果显示,CsMVD在茶树不同组织中均有表达,果实中CsMVD的表达量最高,表达水平表现为:果实>茎>根>叶>花;在白茶萎凋过程中,CsMVD基因的表达量在48 h结束阶段显著高于0 h,推测该基因与茶叶萜类香气化合物形成有关。

    Abstract:

    Based on the transcriptome database previously detected, we cloned the fulllength cDNA sequence of mevalonate diphosphate decarboxylase gene(MVD) using RTPCR technique from the tea plant of Fuding Dabaicha cultivar. The fulllength cDNA was named CsMVD GenBank accession number MF772780. CsMVD is 1 585 bp in length, containing a 1 266 bp ORF encoding 421 amino acid. The predict protein molecular and theoretic isoelectric point of CsMVD are 46.45 kD and 7.10, respectively. CsMVD protein contains an conserved domain of MVD1 superfamily which is primarily located in the cytoplasm. There are multiple phosphorylation sites at CsMVD protein but with no transmembrane structure and signal peptide. Some conservative amino acid residues determine the specific catalytic and activity of the protein. Phylogenetic tree analysis showed that CsMVD had the closest genetic relationship with Arnebia euchroma MVD. Tissue expression analysis showed that CsMVD expressed among five tested tissues and had the highest level in fruits successively followed by stem, root, leaf and flower. The expression of CsMVD was significantly higher than that of 0 h at the end of 48 h in the withering procedure of White tea. These results demonstrated that CsMVD might be correlated to the formation of tea terpene aroma compounds.

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王鹏杰,陈 丹,曹红利,等.茶树甲羟戊酸焦磷酸脱羧酶基因CsMVD的克隆与表达分析[J].西北植物学报,2017,37(12):2342-2349

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  • 在线发布日期: 2017-12-29
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