四川农业大学学报 ›› 2019, Vol. 37 ›› Issue (01): 22-27.doi: 10.16036/j.issn.1000-2650.2019.01.004

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金线莲辣椒红素/玉红素合成酶基因CCS的克隆及表达模式分析

刘柄良1, 杨琳1,2, 付凤玲1, 李晚忱1, 张君诚2, 于好强1,*   

  1. 1.四川农业大学玉米研究所/农业部西南玉米生物学与遗传育种重点实验室,成都 611130;
    2.三明学院,福建 三明 365000
  • 收稿日期:2018-01-29 出版日期:2019-02-28 发布日期:2021-01-29
  • 作者简介:刘柄良,博士研究生。*责任作者:于好强,博士,讲师,主要从事植物分子生物学、玉米非生物逆境遗传与改良研究工作,E-mail:yhq1801@sicau.edu.cn。
  • 基金资助:
    福建省教育厅中青年教师教育科研基金项目(JA15470)

Cloning and Expression Analysis of CCS Gene in Anoectochilus

LIU Bingliang1, YANG Lin1,2, FU Fengling1, LI Wanchen1, ZHANG Juncheng2, YU Haoqiang1,*   

  1. 1. Maize Research Institute of Sichuan Agricultural University/Key Laboratory of Maize Biology and Genetic Breeding in Southwest,Ministry of Agriculture,Chengdu 611130,China;
    2. Sanming University,Sanming 365000,Fujian China
  • Received:2018-01-29 Online:2019-02-28 Published:2021-01-29

摘要: 【目的】克隆金线莲辣椒红素/玉红素合成酶(capsanthin/capsorubin synthase,CCS)编码基因并分析其功能。【方法】在金线莲转录组测序数据基础上,同源克隆福建金线莲(Anoectochilus roxburghii)和台湾金线莲(Anoectochilus formasanus)的CCS基因编码序列,并进行生物信息学分析和表达特性分析。【结果】福建金线莲ArCCS基因的ORF长1 461 bp,编码486个氨基酸。台湾金线莲AfCCS 基因的ORF长1 458 bp,编码485个氨基酸。ArCCSAfCCS蛋白序列相似度为97%,均定位于叶绿体,二者理化性质和三级结构高度相似。ArCCSAfCCS基因的表达显著受盐胁迫和不同光质的诱导或抑制。AfCCS基因在根中低表达,在茎中高表达,而ArCCS基因的表达无组织特异性。【结论】成功克隆到福建金线莲和台湾金线莲的CCS基因,二者的表达受到环境因素的显著诱导。

关键词: 金线莲, 辣椒合酶素, 基因克隆, 表达模式, 胁迫

Abstract: 【Objective】 Cloning and functional characterization of the capsanthin/capsorubin synthase (CCS) gene from Anoectochilus. 【Method】 On the basis of the RNA sequencing results,the coding sequence of CCS gene from Anoectochilus roxburghii and Anoectochilus formasanus was cloned by homology-based cloning and analyzed through bioinformatics. The expression pattern of the ArCCS and AfCCS gene was analyzed by quantitative real time PCR. 【Result】 The open reading frame (ORF) of the ArCCS gene was 1 461 bp in length,encoding 486 amino acids. The ORF of the AfCCS gene was 1 458 bp in length,encoding 485 amino acids. The identity between ArCCS and AfCCS protein was 97%. The two proteins localized on the chloroplast,and showed a high similarity on the physical and chemical properties and tertiary structure. The expression of the ArCCS and AfCCS gene was significantly induced or inhibited by the salt stress and different light quality. The AfCCS gene dominantly expressed in stem and weakly expressed in roots. But the ArCCS gene exhibited no tissue-specific expression.【Conclusion】 The sequences of the CCS gene from Anoectochilus roxburghii and Anoectochilus formasanus were successfully cloned,and the expression of the both genes was significantly affected by environmental stimuli.

Key words: Anoectochilus, capsanthin-capsorubin synthase, gene cloning, expression pattern, stress

中图分类号: 

  • S682.31