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植物研究 ›› 2016, Vol. 36 ›› Issue (4): 556-564.doi: 10.7525/j.issn.1673-5102.2016.04.011

• 研究报告 • 上一篇    下一篇

油茶柠檬酸合成酶(CS)基因的克隆和表达分析

叶思诚, 姚小华, 王开良, 林萍, 龚洪恩, 卓仁英   

  1. 中国林业科学研究院亚热带林业研究所, 富阳 311400
  • 收稿日期:2016-03-16 出版日期:2016-07-15 发布日期:2016-06-15
  • 通讯作者: 姚小华,E-mail:yaoxh168@163.com E-mail:yaoxh168@163.com
  • 作者简介:叶思诚(1985-),男,博士研究生,主要从事经济林栽培育种研究。
  • 基金资助:
    浙江省重大农业专项"油茶高产优质新品种选育及示范"(2012C12908)

Cloning and Expression Analysis of Citrate Synthase(CS) Gene in Camellia oleifera

YE Si-Cheng, YAO Xiao-Hua, WANG Kai-Liang, LIN Ping, GONG Hong-En, ZHUO Ren-Ying   

  1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400
  • Received:2016-03-16 Online:2016-07-15 Published:2016-06-15
  • Supported by:
    This work was supported by Zhejiang Agriculture Major Project ‘Breeding and demonstration of the new cultivars of Camellia oleifera with high yield and good quality’(2012C12908)

摘要: 采用RT-PCR技术从油茶中分离出一个柠檬酸合成酶基因,该基因的cDNA全长1416 bp,编码471个氨基酸,推导的蛋白分子量为52.74 kD,理论等电点(PI)为6.95。同源比对显示其与其他植物的CS蛋白序列高度同源,将该基因命名为CoCS(Gen Bank登录号:KU161147)。系统进化树分析表明油茶CoCS与杜鹃和葡萄的CS蛋白的亲缘关系较近。荧光定量PCR分析结果表明,油茶受到低磷胁迫后根系CoCS基因的表达受到低磷诱导,表达量呈现先升高后降低的趋势;不同油茶品种不同组织(根、茎、叶)中的CoCS基因在不用磷处理下的表达模式不同。

关键词: 油茶, 柠檬酸合成酶, 低磷, 基因克隆, 表达分析

Abstract: A citrate synthase(CS) gene were isolated from Camellia oleifera by RT-PCR. The full-length cDNA of the CS is 1416 bp in size, encodeding a deduced polypetide of 471 amino acids with estimated molecular weight of 52.74 kD and theoretical isoelectric point of 6.95. Homologous alignment showed that the deduced protein had high identities with the CS proteins of other plants, therefore the gene was named as CoCS(Genbank No.KU161147). By phylogenetic tree analysis, CoCS had close genetic relationships with Rhododendron micranthum and Vitis vinifera. By qRT-PCR, the expression of CoCS in root was induced by phosphate deficiency and increased at first and decreased subsequently. The expression patterns of CoCS were different among different tissues and different cultivars.

Key words: Camellia oleifera, citrate synthase, phosphate deficient, gene cloning, expression analysis

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