Staal Jens, Kaliff Maria, Dewaele Ellen, Persson Mattias, Dixelius Christina
Department of Molecular Biomedical Research, Unit for Molecular Signal Transduction in Inflammation, VIB, Ghent University, Technologiepark 927, Ghent (Zwijnaarde) B-9052, Belgium.
Plant J. 2008 Jul;55(2):188-200. doi: 10.1111/j.1365-313X.2008.03503.x. Epub 2008 Apr 4.
Here, we describe the rapid cloning of a plant gene, Leptosphaeria maculans 3 (RLM3(Col)), which encodes a putative Toll interleukin-1 receptor-nucleotide binding (TIR-NB) class protein, which is involved in defence against the fungal pathogen L. maculans and against three other necrotrophic fungi. We have, through microarray-based case control bulk segregant comparisons of transcriptomes in pools of Col-0 x An-1 progeny, identified the absence of a locus that causes susceptibility in An-1. The significance of this locus on chromosome 4 for L. maculans resistance was supported by PCR-based mapping, and denoted resistance to RLM3(Col). Differential susceptible phenotypes in four independent T-DNA insertion lines support the hypothesis that At4g16990 is required for RLM3(Col) function. The mutants in RLM3(Col) also exhibited an enhanced susceptibility to Botrytis cinerea, Alternaria brassicicola and Alternaria brassicae. Complementations of An-1 and T-DNA mutants using overexpression of a short transcript lacking the NB-ARC domain, or a genomic clone, restored resistance to all necrotrophic fungi. The elevated expression of RLM3(Col) on B. cinerea-susceptible mutants further suggested convergence in signalling and gene regulation between defence against B. cinerea and L. maculans. In the case of L. maculans, RLM3(Col) is required for efficient callose deposition downstream of RLM1(Col).
在此,我们描述了一种植物基因——大茎点菌3(RLM3(Col))的快速克隆,该基因编码一种假定的Toll样白细胞介素-1受体-核苷酸结合(TIR-NB)类蛋白,它参与对真菌病原体大茎点菌以及其他三种坏死营养型真菌的防御。我们通过基于微阵列的Col-0×An-1后代群体转录组病例对照混合分离分析,确定了An-1中一个导致感病性的位点缺失。基于PCR的定位支持了4号染色体上该位点对大茎点菌抗性的重要性,并将其命名为对RLM3(Col)的抗性。四个独立的T-DNA插入系中的不同感病表型支持了At4g16990是RLM3(Col)功能所必需的这一假设。RLM3(Col)突变体对灰葡萄孢、芸苔链格孢和芸苔链格孢也表现出增强的感病性。使用缺乏NB-ARC结构域的短转录本或基因组克隆进行过表达对An-1和T-DNA突变体进行互补,恢复了对所有坏死营养型真菌的抗性。RLM3(Col)在灰葡萄孢易感突变体上的高表达进一步表明,对灰葡萄孢和大茎点菌的防御在信号传导和基因调控方面存在趋同。就大茎点菌而言,RLM3(Col)是RLM1(Col)下游有效胼胝质沉积所必需的。