The Identification of Genes Important in Pseudomonas syringae pv. phaseolicola Plant Colonisation Using In Vitro Screening of Transposon Libraries.
The Identification of Genes Important in Pseudomonas syringae pv. phaseolicola Plant Colonisation Using In Vitro Screening of Transposon Libraries.
复制标题
鉴定在丁香假单胞菌PV中重要的基因。使用转座子文库的体外筛选阶段植物定植。
DOI：
10.1371/journal.pone.0137355
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Arnold DL
中科院分区：
文献类型：
--
作者：
Manoharan B;Neale HC;Hancock JT;Jackson RW;Arnold DL
细菌性植物病原菌丁香假单胞菌致病变种。菜豆属（Phaseolicola，Pph）在通过伤口和气孔进入植物组织内部之前定殖于普通豆类植物的表面。在细胞间隙中，病原体在质外体液中增殖，并在植物细胞周围形成小菌落（生物膜）。如果病原体能够抑制植物的自然抗性反应，就会引起晕轮疫病。抗性抑制的过程是相当好理解的，但病原体在定殖中使用的机制尚不清楚。我们假设我们可以应用体外遗传筛选来寻找运动性、集落形成和粘附的变化，这些变化是感染、小集落形成和细胞粘附的代表。我们构建了Pph菌株1448 A和1302 A的转座子（Tn）突变体文库，发现106/1920突变体表现出菌落形态、运动性和生物膜形成的改变。如预期的那样，在基因组内鉴定的Tn的插入点的鉴定突出显示了一些在编码鞭毛的各个部分的基因中具有突变的改变的运动性突变体。还鉴定了参与营养物质生物合成的基因、膜相关蛋白和一些保守的假设蛋白（CHP）基因。一个CHP基因的突变导致了植物中细菌生长的正向增加。这种快速和廉价的筛选方法允许在体外性状，可以相关的植物相互作用中的作用的基因的发现。
The bacterial plant pathogen Pseudomonas syringae pv. phaseolicola (Pph) colonises the surface of common bean plants before moving into the interior of plant tissue, via wounds and stomata. In the intercellular spaces the pathogen proliferates in the apoplastic fluid and forms microcolonies (biofilms) around plant cells. If the pathogen can suppress the plant’s natural resistance response, it will cause halo blight disease. The process of resistance suppression is fairly well understood, but the mechanisms used by the pathogen in colonisation are less clear. We hypothesised that we could apply in vitro genetic screens to look for changes in motility, colony formation, and adhesion, which are proxies for infection, microcolony formation and cell adhesion. We made transposon (Tn) mutant libraries of Pph strains 1448A and 1302A and found 106/1920 mutants exhibited alterations in colony morphology, motility and biofilm formation. Identification of the insertion point of the Tn identified within the genome highlighted, as expected, a number of altered motility mutants bearing mutations in genes encoding various parts of the flagellum. Genes involved in nutrient biosynthesis, membrane associated proteins, and a number of conserved hypothetical protein (CHP) genes were also identified. A mutation of one CHP gene caused a positive increase in in planta bacterial growth. This rapid and inexpensive screening method allows the discovery of genes important for in vitro traits that can be correlated to roles in the plant interaction.