中国农业科技导报 ›› 2019, Vol. 21 ›› Issue (12): 102-109.DOI: 10.13304/j.nykjdb.2019.0231

• 动植物健康 • 上一篇    下一篇

小麦CBL结合蛋白激酶TaCIPK31负调控TcLr37对小麦叶锈菌的抗性

张娜1,赵晨光1,温晓蕾2,杨文香1,张娜1*,刘大群1*   

  1. 1.河北农业大学植物保护学院, 河北省农作物病虫害生物防治工程技术研究中心, 国家北方山区农业工程技术研究中心, 河北 保定 071000; 2.河北科技师范学院农学与生命科技学院, 河北 秦皇岛 066000
  • 收稿日期:2019-03-27 出版日期:2019-12-15 发布日期:2019-05-08
  • 通讯作者: *通信作者 张娜 E-mail:zn0318@126.com;刘大群 E-mail:ldq@hebau.edu.cn
  • 作者简介:张娜 E-mail: 530061790@qq.com
  • 基金资助:
    河北省教育厅青年基金项目(2018102021888);国家自然科学基金项目(31871915)。

A CBL-Interacting Protein Kinase Gene TaCIPK31 Negative Regulate the Resistance to Puccinia triticina in Wheat TcLr37

ZHANG Na1, ZHAO Chenguang1, WEN Xiaolei2, YANG Wenxiang1, ZHANG Na1*, LIU Daqun1*   

  1. 1.Biological Control Center of Plant Diseases and Plant Pests of Hebei Province; National Engineering Research Center for Agriculture in Northern Mountainous Areas, College of Plant Protection, Hebei Agricultural University;  Hebei Baoding 071001, China; 2.Hebei Normal University of Science & Technology, Hebei Qinhuangdao 066000, China
  • Received:2019-03-27 Online:2019-12-15 Published:2019-05-08

摘要: CIPK是植物特有的丝氨酸-苏氨酸蛋白激酶家族中的一类SnRK3激酶 (SNF1-related protein kinase3),一般与类钙调磷酸酶亚基B类蛋白CBL(calcineurin B-like protein)相互作用形成信号网络,从而在钙信号介导的生理生化过程中发挥作用。克隆获得小麦中CIPK基因家族的TaCIPK31,其全长1 350 bp,编码449个氨基酸,包含保守的激酶催化结构域及调控结构域,与已报道的粗山羊草、水稻等CIPK蛋白具有高度相似性。定量分析结果表明,TaCIPK31在小麦-叶锈菌亲和反应中显著上调诱导表达。烟草亚细胞定位表明该基因分布于整个细胞。通过病毒诱导的基因沉默研究发现沉默TaCIPK31后小麦材料TcLr37对叶锈菌毒性小种THTT抗性增强。此外,详细的组织学分析表明,TaCIPK31沉默植株中推迟了病原菌在寄主中的定殖。推测TaCIPK31在小麦抵抗叶锈菌侵染过程中起负调控作用,该结果对揭示CIPK蛋白激酶在小麦-叶锈菌互作体系的作用及分子机理提供参考。

关键词: 小麦叶锈病;TaCIPK31;病毒诱导的基因沉默, 抗性

Abstract: CIPK is a plant-specific serine-threonine protein kinase family belonged to the class of SnRK3 kinase (SNF1-related protein kinase3, SnRK3), which mainly interacts with the calcineurin B-like protein (calcineurin B-like protein) to mediate  plant responses to external stimuli and regulate a wide range of physiological processes. This study obtained a CIPK gene, TaCIPK31, from the wheat leaves infected by Puccinia triticinia (Pt). The full-length cDNA of TaCIPK31 was 1 350 bp, which encoding 449 amino acids. Multi-sequence alignment showed that TaCIPK31 shared high similarity with CIPK in Oryza sativa and Aegilops tauschii, etc. Analysis of the protein domain indicated that TaCIPK31 contained conserved regulatory domain  and protein kinases domain. qRT-PCR assays indicated that TaCIPK31 was induced by Pt infection and significantly expressed during compatible interactions between wheat and Pt. Subcellular localization assays revealed that TaCIPK31 showed similar to that of the GFP control, indicating a cytoplasmic and nuclear localization. Silencing of TaCIPK31 in TcLr37  induced by virus  enhanced resistance of wheat to the virulent Pt isolate THTT. Moreover, detailed histological analysis showed that silencing of TaCIPK31 delayed the colonization of the haustorial mother cell in the host. These results suggested that TaCIPK31 negatively regulated wheat resistance to Pt infection, which provided a reference for revealing the role and molecular mechanism of CIPK protein kinase in wheat-leaf rust interaction system.

Key words: wheat leaf rust disease, TaCIPK31, virus-induced gene silencing, resistance