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首页> 外文期刊>Applied Microbiology >Forward Genetic In Planta Screen for Identification of Plant-Protective Traits of Sphingomonas sp. Strain Fr1 against Pseudomonas syringae DC3000
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Forward Genetic In Planta Screen for Identification of Plant-Protective Traits of Sphingomonas sp. Strain Fr1 against Pseudomonas syringae DC3000

机译：在植物筛选中的正向遗传鉴定鞘氨醇单胞菌的植物保护性状。抵抗丁香假单胞菌DC3000的Fr1菌株

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Sphingomonas sp. strain Fr1 has recently been shown to protect Arabidopsis thaliana against the bacterial leaf pathogen Pseudomonas syringae DC3000. Here, we describe a forward genetic in planta screen to identify genes in Sphingomonas sp. Fr1 necessary for this effect. About 5,000 Sphingomonas sp. Fr1 mini-Tn 5 mutants were assayed for a defect in plant protection against a luxCDABE -tagged P. syringae DC3000 derivative in a space-saving 24-well plate system. The bioluminescence of the pathogen was used as the indicator of pathogen proliferation and allowed for the identification of Sphingomonas sp. Fr1 mutants that had lost the ability to restrict pathogen growth before disease symptoms were visible. Potential candidates were validated using the same miniaturized experimental system. Of these mutants, 10 were confirmed as plant protection defective yet colonization competent. The mutants were subsequently evaluated in a previously described standard microbox system, and plants showed enhanced disease phenotypes after pathogen infection relative to those inoculated with the parental strain as a control. However, the disease severities were lower than those observed for control plants that were grown axenically prior to pathogen challenge, which suggests that several traits may contribute to plant protection. Transposon insertion sites of validated mutants with defects in plant protection were determined and mapped to 7 distinct genomic regions. In conclusion, the established screening protocol allowed us to identify mutations that affect plant protection, and it opens the possibility to uncover traits important for in planta microbe-microbe interactions.
机译：鞘氨醇单胞菌Fr1菌株最近显示出可以保护拟南芥免受细菌叶病原体丁香假单胞菌DC3000的侵害。在这里，我们描述了植物筛选中的正向遗传学，以鉴定鞘氨醇单胞菌sp。中的基因。为此需要的Fr1。约5,000只鞘氨醇单胞菌在节省空间的24孔板系统中检测了Fr1 mini-Tn 5突变体在针对luxCDABE标记的丁香假单胞菌DC3000衍生物的植物保护中的缺陷。病原体的生物发光用作病原体增殖的指标，并用于鉴定鞘氨醇单胞菌。 Fr1突变体已经失去了限制病原体生长的能力，使疾病症状变得可见。使用相同的小型实验系统验证了潜在的候选对象。在这些突变体中，有10个被确认为植物保护缺陷但具有定植能力。随后在先前描述的标准微型盒系统中评估突变体，并且相对于接种亲本菌株作为对照的植物，植物在病原体感染后表现出增强的病害表型。但是，病害严重程度低于在病原体攻击之前先进行了轴心培养的对照植物，这表明某些性状可能有助于植物保护。确定具有植物保护缺陷的经过验证的突变体的转座子插入位点，并将其定位到7个不同的基因组区域。总之，已建立的筛选方案使我们能够鉴定出影响植物保护的突变，这为揭示对植物微生物与微生物相互作用重要的性状提供了可能性。

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