小麦CBL结合蛋白激酶TaCIPK31负调控TcLr37对小麦叶锈菌的抗性

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。

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

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.