草地学报

草地学报 ›› 2016, Vol. 24 ›› Issue (2): 447-452.DOI: 10.11733/j.issn.1007-0435.2016.02.030

• 研究专报 • 上一篇    下一篇

结缕草ZjCCS基因的克隆与表达分析

樊波1, 孙鑫博2, 张胤冰1, 张雪1, 袁建波1, 张海兰1, 肖维阳1, 韩烈保1, 许立新1   

  1. 1. 北京林业大学草坪研究所, 北京 100083;
    2. 河北省作物生长调控重点实验室 河北农业大学, 河北 保定 071001
  • 收稿日期:2015-10-12 修回日期:2016-02-01 出版日期:2016-04-15 发布日期:2016-06-06
  • 通讯作者: 许立新
  • 作者简介:樊波(1990-),女,内蒙古呼和浩特人,硕士研究生,研究方向为草坪草分子育种,E-mail:fanbobecky@163.com
  • 基金资助:

    国家高新技术研究发展计划项目(2013AA102607)资助

Cloning and Expression Analysis of ZjCCS from Zoysia Japonica

FAN Bo1, SUN Xin-bo2, ZHANG Yin-bing1, ZHANG Xue1, YUAN Jian-bo1, ZHANG Hai-lan1, XIAO Wei-yang1, HAN Lie-bao1, XU Li-xin1   

  1. 1. Institute of Turfgrass Science, Beijing Forestry University, Beijing 100083, China;
    2. Key Laboratory of Crop Growth Regulation of Hebei Province, Agricultural University of Hebei, Baoding, Hebei Province 071001, China
  • Received:2015-10-12 Revised:2016-02-01 Online:2016-04-15 Published:2016-06-06

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摘要：

为了解Cu/Zn超氧化物歧化酶分子伴侣基因(CCS)在结缕草逆境胁迫过程中的表达调控机制,以结缕草(Zoysia japonica)盐胁迫条件下的转录组数据为依据,结缕草cDNA为模板,通过RT-PCR法,克隆了924 bp长的结缕草ZjCCS基因的CDS序列。同源性分析显示该基因与谷子(Setaria italica)同源性最高,含有植物CCS蛋白所特有的3个保守结构域。利用实时荧光定量PCR(qRT-PCR)技术研究该基因在非生物胁迫条件下在结缕草叶片中的表达情况,结果显示,在盐胁迫条件下,ZjCCS基因的表达量呈先上升后下降的趋势,4 h表达量最高,相对于盐胁迫,干旱和重金属胁迫对ZjCCS基因的表达量影响不大,推测ZjCCS基因可能在结缕草响应盐胁迫过程中发挥作用。

关键词: 结缕草, CCS, 非生物胁迫, 实时荧光定量PCR

Abstract:

A chaperone gene (ZjCCS) of copper superoxide dismutase gene of Zoysia japonica was cloned by RT-PCR. The ZjCCS gene contains a 924-nucleotides-long open reading frame(ORF), which encoded a protein of 307 amino acids. Its expression level was analyzed for elucidating its role in response to salt drought and heavy-metal stress. Homology Blast (BlastP) with the deduced amino acids sequence indicated that ZjCCS gene had a high similarity to other plant CCS genes, and the predicted ZjCCS polypeptide contained 3 distinct domains. QRT-PCR experiments showed that ZjCCS mRNA level in leaf was significantly induced by salt stress, and reached to peak after 4 h of salt treatment. Drought stress and heavy-metal stresses didn't have a significant influence on the ZjCCS expression level, which suggested that ZjCCS might play a major role when zoysia japonica responding to salt stress.

Key words: Zoysia japonica, CCS, Abiotic stress, qRT-PCR

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