Chen Kegui, Du Liqun, Chen Zhixiang
Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844-3052, USA.
Plant Mol Biol. 2003 Sep;53(1-2):61-74. doi: 10.1023/B:PLAN.0000009265.72567.58.
During the search for potential target genes of WRKY DNA-binding transcription factors, we have previously identified four pathogen-induced Arabidopsis genes (CRK5, CRK6, CRK10 and CRK11) encoding receptor-like protein kinases (RLKs) containing novel cysteine-rich repeats in their extracellular domains. In the present study, we transformed Arabidopsis plants with the RLK genes under control of the constitutive CaMV 35S promoter or a steroid-inducible Ga14 promoter. Expression of CRK5, but not the three other RLK genes, resulted in significant alterations in defense responses and leaf growth in transgenic plants. In transgenic plants harboring the 35S::CRK5 construct, significantly elevated and constitutive expression of CRK5 correlated with enhanced leaf growth and increased resistance to the bacterial pathogen Pseudomonas syringae. The enhanced disease resistance in the transgenic plants was associated with more rapidly induced expression of the PR1 gene after pathogen infection. In transgenic plants transformed with CRK5 under control of the steroid-inducible promoter, expression of the transgene was induced at relatively high levels after the steroid application and this induced expression of CRK5 triggered hypersensitive response-like cell death. Induced CRK5 expression also activated cell death in the npr1, ndr1 and eds1 mutants and in the transgenic nahG plants that fail to accumulate salicylic acid. Thus, the novel RLK is capable of activating multiple distinct defense responses depending on the manner and/or the levels of its over-expression in transgenic plants.
在寻找WRKY DNA结合转录因子的潜在靶基因过程中,我们之前已鉴定出四个病原体诱导的拟南芥基因(CRK5、CRK6、CRK10和CRK11),它们编码受体样蛋白激酶(RLK),其胞外结构域含有新的富含半胱氨酸的重复序列。在本研究中,我们用组成型CaMV 35S启动子或类固醇诱导型Ga14启动子控制下的RLK基因转化拟南芥植株。CRK5的表达,而非其他三个RLK基因的表达,导致转基因植株的防御反应和叶片生长发生显著改变。在含有35S::CRK5构建体的转基因植株中,CRK5的显著升高和组成型表达与叶片生长增强和对细菌病原体丁香假单胞菌的抗性增加相关。转基因植株中增强的抗病性与病原体感染后PR1基因更快诱导表达有关。在用类固醇诱导型启动子控制下的CRK5转化的转基因植株中,类固醇处理后转基因的表达在相对较高水平被诱导,并且这种CRK5的诱导表达引发了类似过敏反应的细胞死亡。诱导的CRK5表达还激活了npr1、ndr1和eds1突变体以及不能积累水杨酸的转基因nahG植株中的细胞死亡。因此,这种新的RLK能够根据其在转基因植株中的过表达方式和/或水平激活多种不同的防御反应。