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作物学报 ›› 2013, Vol. 39 ›› Issue (04): 599-608.doi: 10.3724/SP.J.1006.2013.00599

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

茶树赤霉素受体基因CsGID1a的克隆与表达分析

岳川1,2,**,曾建明1,**,曹红利1,2,郝心愿1,3,章志芳1,王新超1,*,杨亚军1,*   

  1. 1中国农业科学院茶叶研究所 / 国家茶树改良中心,浙江杭州 310008; 2中国农业科学院研究生院, 北京 100081; 3西北农林科技大学园艺学院,陕西杨凌 712100
  • 收稿日期:2012-09-16 修回日期:2012-12-11 出版日期:2013-04-12 网络出版日期:2013-01-28
  • 通讯作者: 王新超, E-mail: xcw75@mail.tricaas.com, Tel: 0571-86653162; 杨亚军, E-mail: yyjang@mail.tricaas.com
  • 基金资助:

    本研究由国家自然科学基金项目(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 Gibberellin Receptor Gene CsGID1a in Tea Plant (Camellia sinensis)

YUE Chuan1,2,**,ZENG Jian-Ming1,**,CAO Hong-Li1,2,HAO Xin-Yuan1,3,ZHANG Zhi-Fang1,WANG Xin-Chao1,*,YANG Ya-Jun1,*   

  1. 1 National Center for Tea Improvement, Tea Research Institute of Chinese Academy of Agricultural Sciences, Hangzhou 310008, China; 2 Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 College of Horticulture, Northwest A&F University, Yangling 712100, China
  • Received:2012-09-16 Revised:2012-12-11 Published:2013-04-12 Published online:2013-01-28
  • Contact: 王新超, E-mail: xcw75@mail.tricaas.com, Tel: 0571-86653162; 杨亚军, E-mail: yyjang@mail.tricaas.com

摘要:

GID1作为赤霉素信号转导的受体,在赤霉素作用中具有重要作用。采用同源克隆方法,利用RT-PCR和RACE技术在茶树中克隆到赤霉素受体基因GID1的cDNA全长,命名为CsGID1a (GenBank登录号为JX235369)。该基因全长1411 bp,开放阅读框1 023 bp,编码341个氨基酸。生物信息学分析显示,CsGID1a编码的蛋白分子量为38.53 kD,理论等电点为5.62;无信号肽位点,是非分泌性蛋白,具有1个跨膜区,基因被定位于细胞核内;CsGID1a氨基酸序列具有激素敏感性脂肪酶(HSL)家族蛋白的HGG、GXSXG功能域以及羧酸酯酶典型的三级结构;与其他物种的GID1相似性均在60%以上,与葡萄的相似性最大达87%、进化关系最近。荧光定量PCR结果显示,高浓度(1.0×10–5 mol L–1)GA3能够下调CsGID1a的表达,5 h内的表达呈下降趋势;随着越冬茶芽萌动进程,CsGID1a表达量逐渐降低,特别在3月初萌发以后变化较大,推测赤霉素及其受体基因可能与茶树越冬芽解休眠相关。

关键词: 茶树, 赤霉素, GID1, 表达分析, 茶芽解休眠

Abstract:

GID1 (Gibberellin insensitive dwarf1), as the soluble gibberellin (GA) receptor in GA signaling pathway, plays the vital role in GA reactions. In this study, the homologous gene of GID1 was isolated with RT-PCR and RACE-PCR from tea plant (Camellia sinensis). The obtained cDNA sequence, named CsGID1a, had the full-length of 1 411 bp containing a 1 023 bp open reading frame (ORF), encoding 341 amino acid residues, and was submitted to GenBank with accession number JX235369. The bioinformatics characterization indicated that CsGID1a was a non-secretory protein without a signal peptide. The molecular weight and theoretic isoelectric point of CsGID1a are 38.53 kD and 5.62, respectively. CsGID1a was located in the nucleus, encoding a protein with one transmembrane domain. CsGID1a contained hormone sensitive lipsase family (HSL) conserved domains, HGG and GXSXG motif, and shared the plant carboxylesterase tertiary structure. Homologous alignment and phylogenetic tree showed that CsGID1a shared over 60% amino acid sequence similarity with that of other species, and had the highest similarity (87%) and the closest genetic relationship to Vitis vinifera. The real-time PCR analysis showed that the expression of CsGID1a was down-regulated by high concentration of GA3 (1.0×10–5 mol L–1) and reduced slowly during the treatment for five hours. The further experiments suggested that the expression of CsGID1a was also decreased in the process of bud sprouting. These results demonstrated that CsGID1a and GA could be associated with bud bursting in tea plant in spring.

Key words: Tea plant (Camellia sinensis), Gibberellin (GA), GID1, Expression analysis, Bud bursting

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