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作物学报 ›› 2013, Vol. 39 ›› Issue (03): 389-397.doi: 10.3724/SP.J.1006.2013.00389

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

茶树生长素响应因子基因CsARF1的克隆与表达分析

郝心愿1,2,曹红利2,杨亚军2,*,王新超2,*,马春雷2,肖斌1   

  1. 1 西北农林科技大学园艺学院，陕西杨凌 712100；2 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
  • 收稿日期:2012-07-20 修回日期:2012-11-05 出版日期:2013-03-12 网络出版日期:2013-01-04
  • 通讯作者: 杨亚军, E-mail: yjyang@mail.tricaas.com; 王新超, E-mail: xcw75@mail.tricaas.com, Tel: 0571-86653162
  • 基金资助:

    本研究由国家自然科学基金项目(30872059, 31170650), 浙江省自然科学基金重点项目(Z3100473)和国家现代农业产业技术体系建设专项(CARS-23)资助。

Cloning and Expression Analysis of Auxin Response Factor Gene (CsARF1) in Tea Plant (Camellia sinensis [L.] O. Kuntze)

HAO Xin-Yuan1,2,CAO Hong-Li2,YANG Ya-Jun2,*,WANG Xin-Chao2,*,MA Chun-Lei2,XIAO Bin1   

  1. 1 College of Horticulture, Northwest A&F University, Yangling 712100, China; 2 Tea Research Institute of Chinese Academy of Agricultural Sciences, National Center for Tea Improvement, Key Laboratory of Tea Plant Biology and Resources Utilization, Ministry of Agriculture, Hangzhou 310008, China
  • Received:2012-07-20 Revised:2012-11-05 Published:2013-03-12 Published online:2013-01-04
  • Contact: 杨亚军, E-mail: yjyang@mail.tricaas.com; 王新超, E-mail: xcw75@mail.tricaas.com, Tel: 0571-86653162

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被引次数

9

摘要：

生长素响应因子(ARFs)是能够与生长素原初反应基因启动子区的生长素响应基元(TGTCTC)特异结合的一类转录因子，调控生长素应答基因的表达。采用SMART-RACE-PCR技术，获得茶树生长素响应因子基因CsARF1的全长cDNA序列(GenBank登录号为JX307853)，并进行了生物信息学分析和表达分析。CsARF1基因cDNA全长3222 bp，包含2463 bp的开放阅读框(ORF)，编码820个氨基酸残基。编码蛋白的分子量为49.35 kD，具有保守的N端DNA结合域B3和C端二聚化结构域IAA_ARF，中间区域富含谷氨酸、丝氨酸和亮氨酸，是一个定位于细胞质的具有激活转录功能的可溶性蛋白。相似性及系统进化分析表明，该基因编码的氨基酸序列与葡萄ARF8的相似性最高(83%)，与番茄ARF8的亲缘关系最近。利用实时荧光定量PCR技术，检测该基因在茶树越冬芽休眠到萌发后不同时期的表达情况。结果显示CsARF1在茶树越冬芽深休眠和萌动期表达量较高，表明该基因与茶树越冬芽的休眠维持及解除密切相关。

关键词: 茶树, 休眠, 生长素响应因子, 表达分析

Abstract:

Auxin response factors (ARFs) are transcription factors that bind to TGTCTC auxin response elements in promoters of early/primary response genes and regulate the expression of auxinresponse genes. The full-length cDNA of one ARF gene named CsARF1 (GenBank accession number JX307853) was firstly cloned from tea plant (Camellia sinensis [L.] O. Kuntze) by SMART-RACE-PCR. The results indicated that the full-length cDNA of CsARF1 was 3222 bp containing 2463 bp ORF which encoded 820 amino acid residues with a putative molecular mass of 49.35 kD. The soluble protein encoded by CsARF1 functioned in the cytoplasm and consisted of an amino-terminal DNA-binding domain (B3), a carboxy-terminal dimerization domain (IAA_ARF), and a Gln, Ser and Leu-rich middle region, which is proposed to function as an activation domain. Blast and phylogenetic analysis showed that the protein encoded by CsARF1 shared the highest identity (83%) with ARF8 in Vitisvinifera, and had close genetic relationship with ARF8 in Solanumlycopersicum. The expression analysis of CsARF1 conducted by qRT-PCT during the different phases of bud dormancy and bud break indicated that CsARF1 had a marked rise in the expression level at deep dormant stage and sprouting stage, demonstrating that CsARF1 is relevant to the regulation of tea plant bud dormancy and bud break.

Key words: Tea plant (Camellia sinensis), Dormancy, Auxin response factor, Expression analysis

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