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植物研究 ›› 2021, Vol. 41 ›› Issue (4): 604-613.doi: 10.7525/j.issn.1673-5102.2021.04.016

• 研究报告 • 上一篇    

水稻OsRPK2基因的克隆及功能初步鉴定

张福臻, 刘方惠, 陈鑫欣, 孙静, 葛荣朝()   

  1. 河北师范大学生命科学学院,石家庄 050024
  • 收稿日期:2020-02-28 出版日期:2021-07-20 发布日期:2021-03-24
  • 通讯作者: 葛荣朝 E-mail:grcgp@sina.com
  • 作者简介:张福臻(1989—),女,硕士研究生,主要从事耐盐相关基因克隆与功能研究。
  • 基金资助:
    国家自然科学基金(30900104);河北省自然科学基金(C2016205158)

Cloning and Functional Analysis of Rice Gene OsRPK2

Fu-Zhen ZHANG, Fang-Hui LIU, Xin-Xin CHEN, Jing SUN, Rong-Chao GE()   

  1. College of Life Science,Hebei Normal University,Shijiazhuang 050024
  • Received:2020-02-28 Online:2021-07-20 Published:2021-03-24
  • Contact: Rong-Chao GE E-mail:grcgp@sina.com
  • About author:ZHANG Fu-Zhen(1989—),female,graduate,mainly engaged in salt-related gene cloning and functional studies.
  • Supported by:
    National Natural Science Fund of China(30900104);Hebei Natural Science Fund(C2016205158)

摘要:

类受体蛋白激酶基因OsRPK1在水稻的抗逆信号传导中起着重要作用。本研究扩增获得与OsRPK1高度同源的OsRPK2基因,构建p1300:35S:OsRPK2过表达载体后转化拟南芥。对35S:OsRPK2纯合体拟南芥进行抗逆性分析表明,在盐、ABA、PEG胁迫下,OsRPK2过表达拟南芥萌发率都明显低于野生型拟南芥,其幼苗的根长生长和成株生长状况方面比野生型拟南芥表现出更为明显的受抑现象。生理检测表明,盐胁迫处理后,与野生型拟南芥相比,35S:OsRPK2转基因拟南芥中叶绿素含量下降更为明显,脯氨酸上升量较小,丙二醛含量上升更为明显,这些内在生理机制使得OsRPK2过表达拟南芥抗逆性明显下降。通过对35S:OsRPK2拟南芥的qRT-PCR检测发现,OsRPK2的过量表达使拟南芥抗逆信号通路下游的SAD、SOS3FRY基因表达明显受到抑制,OsRPK2基因可能通过SOS和CDPK信号通路影响拟南芥的抗逆性。

关键词: OsRPK2, 类受体蛋白激酶, 抗逆性, 生理检测, 信号通路

Abstract:

Receptor-like protein kinase gene OsRPK1 plays an important role in the resistance signal transduction of rice. OsRPK2 gene, which is highly homologous to OsRPK1, was amplified and transformed into Arabidopsisthaliana by the overexpression vector p1300:35S:OsRPK2. The stress resistance analysis of 35S:OsRPK2Arabidopsis homozygous showed that the germination rate of OsRPK2 overexpressed Arabidopsis was significantly lower than that of the wild-type under salt, ABA and PEG stress. The root and plant growth of the OsRPK2 overexpressed Arabidopsis seedlings showed more obvious inhibition than that of wild-type. Physiological detection of 35S:OsRPK2Arabidopsis showed that the chlorophyll content decreased more significantly, proline increased less and malondialdehyde content increased more significantly than the wild-typeafter salt treatment. These physiological mechanisms of OsRPK2 overexpression Arabidopsis significantly reduce the stress resistance. Through qRT-PCR detection of the OsRPK2 overexpressed Arabidopsis, the overexpression of OsRPK2 inhibited the expression of SADSOS3 and FRY genes significantly. Therefore, OsRPK2 gene may affect Arabidopsis resistance through SOS and CDPK signaling pathway.

Key words: OsRPK2, receptor-like protein kinase, stress resistance, physiological detection, signaling pathway

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