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茶叶科学 ›› 2015, Vol. 35 ›› Issue (1): 45-54.doi: 10.13305/j.cnki.jts.2015.01.009

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茶树生长素受体基因CsTIR1的克隆与表达分析

曹红利1,2, 岳川1,2, 周艳华1,2, 王璐1, 郝心愿1, 曾建明1, 杨亚军1,*, 王新超1,*   

  1. 1. 中国农业科学院茶叶研究所/国家茶树改良中心,农业部茶树生物学与资源利用重点实验室,浙江 杭州 310008;
    2. 中国农业科学院研究生院,北京 100081
  • 收稿日期:2014-09-09 修回日期:2014-10-15 出版日期:2015-02-15 发布日期:2019-08-23
  • 通讯作者: *
  • 作者简介:曹红利,女,博士研究生,主要从事茶树抗寒育种和分子生物学研究。
  • 基金资助:
    国家自然科学基金(31370690)、国家茶叶产业技术体系(CARS-23)资助

Cloning and Expression Analysis of Auxin Receptor Gene CsTIR1 in Tea Plant (Camellia sinensis)

CAO Hongli1,2, YUE Chuan1,2, ZHOU Yanhua1,2, WANG Lu1, HAO Xinyuan1, ZENG Jianming1, YANG Yajun1,*, WANG Xinchao1,*   

  1. 1. Tea Research Institute of the Chinese Academy of Agricultural Sciences, National Center for Tea Improvement, Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, Hangzhou 310008, China;
    2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2014-09-09 Revised:2014-10-15 Online:2015-02-15 Published:2019-08-23

摘要: 吲哚-3-乙酸(又称IAA或生长素),在植物生长发育中具有重要的调控作用,其作用主要通过信号转导途径来完成。生长素受体是其信号转导的关键元件之一。本研究通过RACE克隆,获得了茶树中生长素受体CsTIR1基因的cDNA全长序列(NCBI登录号:JX050147)。CsTIR1序列全长2β315βbp,含1β746βbp的完整开放阅读框,编码581个氨基酸,预测分子量65.18βkD,理论等电点(pI)5.64。茶树CsTIR1与烟草TIR1的相似性最高达82%,亲缘关系最近。CsTIR1含有1个F-box结构域和6个LRR结构域,三级结构形如“蘑菇状”。CsTIR1在茶树的根、茎、叶和花中具有组织表达特异性;其表达受IAA诱导,3种不同浓度的IAA均能诱导CsTIR1上调表达,且在50βμmol·L-1浓度下表达量最大;ABA、GA3、MeJA和BR等激素也能够显著上调其表达;CsTIR1在越冬休眠芽中的表达量低,在活跃期中上调表达。

关键词: 茶树, 植物激素, 生长素, 生长素受体基因CsTIR1, 芽休眠

Abstract: Indole-3-acetic acid (IAA or auxin), functioning via its signal transduction, plays a pivotal role in plant growth and development regulation. Transport inhibitor response 1 (TIR1) protein, an auxin receptor, is one of the most critical components in IAA signaling pathway. The full-length cDNA sequence of CsTIR1 gene was obtained by using RACE technique, and submitted to GenBank with accession number JX050147. The CsTIR1 cDNA length was 2β315βbp, and contained a 1β746βbp open reading frame (ORF), encoding 581 amino acid residues. The molecular weight and theoretic isoelectric point of CsTIR1 protein are 65.18 kD and 5.64, respectively. In addition, CsTIR1 protein had the highest sequence similarity about 82% and the closest genetic relationship to Nicotiana tabacum. The CsTIR1 was predicted to contain one F-box and six leucine-rich-repeat (LRR) domains, which forming the ‘stem’ and ‘cap’, respectively. And its tertiary structure is shaped as a mushroom. Semi-quantitative RT-PCR results suggested that CsTIR1 expression showed a tissue-specificity among root, stems, leaves and flowers. The further investigation indicated that the transcript of CsTIR1 was regulated by phytohormones. In a time-course assay, it was found that CsTIR1 was significantly up-regulated when tea plant treated with three different IAA concentration and various plant hormones (ABA, GA3, MeJA and BR), and showed the highest expression level under 50 μmol·L-1 IAA concentration. Finally, the expression of CsTIR1 was detected in bud dormancy-active cycle during winter and CsTIR1 showed a low transcription level in dormant buds but expressed abundantly in active buds.

Key words: tea plant, phytohormone, auxin, auxin receptor gene CsTIR1, bud-dormancy

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