茶树单萜合成酶CsTPS 基因的克隆及表达分析
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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(闽教科〔2015〕75号);


Cloning and Expression of Monoterpene Synthase Gene CsTPS 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‘铁观音’茶树品种的芽叶为供试材料,采用RTPCR技术,克隆了茶树的1条单萜合成酶(TPS)基因全长cDNA序列,命名为CsTPS(GenBank登录号为KY829105)。CsTPS基因全长2 077 bp,其开放阅读框(ORF)为1 752 bp,编码583个氨基酸。CsTPS基因编码蛋白含有单萜合成酶蛋白特有的2个天冬氨酸富集基序及RRx8W、RxR保守基序,N 端具有一段叶绿体转运肽。同源比对分析显示,CsTPS氨基酸序列与多种植物的单萜合成酶序列相似性较高,与黄胡萝卜α松油醇合成酶、白簕柠檬烯合成酶、蓖麻单萜合成酶相似性分别为 74%、71% 和66%。系统进化树分析表明,CsTPS 属于TPSb亚家族类型。荧光定量PCR结果显示,在白茶萎凋、乌龙茶做青、红茶发酵等加工过程中,CsTPS基因的表达量均有上调,推测CsTPS基因的表达与茶叶加工过程中茶叶香气形成密切相关。

    Abstract:

    In this study,an encoding monoterpene synthase gene named as CsTPS was isolated from the transcriptome database with GenBank accession number KY829105,and its fulllength cDNA sequence was cloned using RTPCR technique from the tea plant of Tieguanyin cultivar. CsTPS was 2 077 bp in length,containing a 1 752 bp open reading frame(ORF)which encoded 583 amino acid. It was predicted that CsTPS protein containing two aspartic acid enriched motifs, RRx8W and RxR conserved domains,and a chloroplast transit peptide in the Nterminal region. The homology analysis showed that CsTPS was homologous to the monoterpene synthase sequence of various plants,and had 74%,71% and 66% similarity with Daucus carota subsp. Sativus αterpineol synthase,Eleutherococcus trifoliatus limonene synthase and Ricinus communis monoterpene synthase,respectively. Phylogenetic tree analysis showed that CsTPS and several monoterpene synthase of various plants were clustered into TPSb subfamily. Expression analysis showed that CsTPS was upregulated in the processing of White tea withering,Oolong tea zuoqing and Black tea fermentation. In conclusion, CsTPS might be correlated to aroma formation in the tea processing.

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王鹏杰,陈 丹,俞 滢,等.茶树单萜合成酶CsTPS 基因的克隆及表达分析[J].西北植物学报,2017,37(8):1465-1473

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  • 在线发布日期: 2017-09-20
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