Zhou Huanbin, Li Shaofang, Deng Zhiyong, Wang Xianping, Chen Tao, Zhang Jinsong, Chen Shouyi, Ling Hongqing, Zhang Aimin, Wang Daowen, Zhang Xiangqi
The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Plant J. 2007 Nov;52(3):420-34. doi: 10.1111/j.1365-313X.2007.03246.x. Epub 2007 Aug 30.
Considerable progress has been made in understanding the function of receptor-like kinase (RLK) genes in model plants. However, much less is known about these genes in crop species. Here we report the characterization of three new wheat RLK genes (TaRLK-R1, 2 and 3). The primary structure of the putative proteins TaRLK-R1, 2 and 3 contained a signal peptide, a cysteine-rich extracellular domain, a transmembrane domain, and a predicted intracellular kinase domain. The fusions between TaRLK-R1, 2 or 3 and the green fluorescence protein (GFP) were targeted to the plasma membrane; such targeting required the signal peptide, extracellular domain and transmembrane domain. Transcription of TaRLK-R1, 2 and 3 was found mainly in the green organs, and was regulated by light. Transcript levels of TaRLK-R1, 2 and 3 increased during the hypersensitive reaction (HR) to stripe rust fungus. In addition, the TaRLK-R3 transcript level was also upregulated by abiotic stresses. Further experiments revealed that the recombinant kinase domain of TaRLK-R3 exhibited auto-phosphorylation activity in vitro. Knocking down the transcript levels of TaRLK-R1, 2 or 3 individually or all together by virus-induced gene silencing compromised the wheat HR to stripe rust fungus. The demonstration of TaRLK-R1, 2 and 3 as positive contributors in the wheat HR to stripe rust fungus suggests a new direction for further functional studies of this important family of RLK genes, and may facilitate the breeding of wheat varieties resistant to stripe rust disease.
在了解模式植物中类受体激酶(RLK)基因的功能方面已经取得了相当大的进展。然而,对于作物物种中的这些基因却知之甚少。在此,我们报告了三个新的小麦RLK基因(TaRLK-R1、2和3)的特征。推测的TaRLK-R1、2和3蛋白的一级结构包含一个信号肽、一个富含半胱氨酸的细胞外结构域、一个跨膜结构域和一个预测的细胞内激酶结构域。TaRLK-R1、2或3与绿色荧光蛋白(GFP)的融合蛋白定位于质膜;这种定位需要信号肽、细胞外结构域和跨膜结构域。发现TaRLK-R1、2和3的转录主要发生在绿色器官中,并受光调控。在对条锈菌的过敏反应(HR)过程中,TaRLK-R1、2和3的转录水平升高。此外,TaRLK-R3的转录水平也受到非生物胁迫的上调。进一步的实验表明,TaRLK-R3的重组激酶结构域在体外表现出自磷酸化活性。通过病毒诱导的基因沉默单独或一起降低TaRLK-R1、2或3的转录水平,会损害小麦对条锈菌的HR。TaRLK-R1、2和3作为小麦对条锈菌HR的积极贡献者的证明,为该重要的RLK基因家族的进一步功能研究指明了新方向,并可能有助于抗条锈病小麦品种的培育。