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Reporter-based screen for Arabidopsis mutants compromised in nonhost resistance

机译：非宿主抗性受损的拟南芥突变体的基于记者的筛选

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摘要

Plants are exposed to many potentially pathogenic microbes in the environment, but each species is only susceptible to a limited number of pathogens. The broad resistance is referred to as nonhost resistance. To date, little is known about the underlying mechanism of nonhost resistance and the signaling transduction process. Here we describe a simple method for isolating Arabidopsis nonhost resistance mutants against a nonadapted bacterial pathogen. A RAP2.6 promoter-driven LUC reporter system was developed to replace the tedious bacterial growth assay during the primary screening. The RAP2.6-LUC reporter gene is normally induced by the virulent bacterium Pseudomonas syringae pv tomato but not the nonadapted bacterium P. syringae pv phaseolicola. By using this method we isolated 4 mutants displaying strong reporter activity in response to P. syringae pv phaseolicola, which were characterized in some details. ebs1, ebs2, ebs3, and ebs4 (enhanced bacterial susceptibility) were compromised in resistance against P. syringae pv phaseolicola and/or P. syringae pv tomato. In addition, ebs4 showed enhanced hypersensitive response to the incompatible bacterium P. syringae pv tomato (avrB). These results demonstrated that the method is suited for large scale screening for nonhost resistance mutants.
机译：植物在环境中暴露于许多潜在的病原微生物，但是每种物种仅对有限数量的病原体敏感。广泛的抗性称为非宿主抗性。迄今为止，对非宿主抗性和信号转导过程的潜在机制知之甚少。在这里，我们描述了一种针对不适应的细菌病原菌分离拟南芥非宿主抗性突变体的简单方法。开发了RAP2.6启动子驱动的LUC报告系统，以取代初筛过程中繁琐的细菌生长测定。 RAP2.6-LUC报告基因通常由强力细菌丁香假单胞菌pv番茄诱导，而不由不适应的细菌丁香假单胞菌pv phaseolicola诱导。通过使用此方法，我们分离了4个突变体，这些突变体对丁香假单胞菌pv phaseolicola有较强的报道活性，并在某些细节上进行了表征。 ebs1，ebs2，ebs3和ebs4（增强的细菌敏感性）在对丁香假单胞菌菜豆和/或丁香假单胞菌菜番茄的抗性中受到损害。此外，ebs4对不兼容的丁香假单胞菌pv番茄（avrB）表现出更高的过敏反应。这些结果表明该方法适用于非宿主抗性突变体的大规模筛选。

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