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Statistical Analysis of Pseudomonas aeruginosa Biofilm Development: Impact of Mutations in Genes Involved in Twitching Motility Cell-to-Cell Signaling and Stationary-Phase Sigma Factor Expression

机译:铜绿假单胞菌生物膜发育的统计分析:抽动运动细胞间信号和平稳阶段西格玛因子表达的基因突变的影响。

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摘要

Four strains of Pseudomonas aeruginosa (wild type, ΔpilHIJK mutant, lasI mutant, and rpoS mutant) were genetically tagged with the green fluorescent protein, and the development of flow chamber-grown biofilms by each of them was investigated by confocal laser scanning microscopy. The structural developments of the biofilms were quantified by the computer program COMSTAT (A. Heydorn, A. T. Nielsen, M. Hentzer, C. Sternberg, M. Givskov, B. K. Ersbøll, and S. Molin, Microbiology 146:2395-2407, 2000). Two structural key variables, average thickness and roughness, formed the basis for an analysis of variance model comprising the four P. aeruginosa strains, five time points (55, 98, 146, 242, and 314 h), and three independent rounds of biofilm experiments. The results showed that the wild type, the ΔpilHIJK mutant, and the rpoS mutant display conspicuously different types of temporal biofilm development, whereas the lasI mutant was indistinguishable from the wild type at all time points. The wild type and the lasI mutant formed uniform, densely packed biofilms. The rpoS mutant formed densely packed biofilms that were significantly thicker than those of the wild type, whereas the ΔpilHIJK mutant formed distinct microcolonies that were regularly spaced and almost uniform in size. The results are discussed in relation to the current model of P. aeruginosa biofilm development.
机译:用绿色荧光蛋白基因标记了四株铜绿假单胞菌(野生型,ΔpilHIJK突变体,lasI突变体和rpoS突变体),并通过共聚焦激光扫描显微镜研究了它们各自在流通腔中生长的生物膜的发育。生物膜的结构发育通过计算机程序COMSTAT进行定量(A.Heydorn,AT Nielsen,M.Hentzer,C.Sternberg,M.Givskov,BKErsbøll和S.Molin,Microbiology 146:2395-2407,2000) 。两个结构关键变量(平均厚度和粗糙度)构成了包括四个铜绿假单胞菌菌株,五个时间点(55、98、146、242和314 h)以及三轮独立生物膜的方差模型分析的基础。实验。结果表明,野生型,ΔpilHIJK突变体和rpoS突变体表现出明显不同的时间生物膜发育类型,而lasI突变体在所有时间点都与野生型没有区别。野生型和lasI突变体形成均匀,密集的生物膜。 rpoS突变体形成了紧密堆积的生物膜,比野生型膜厚得多,而ΔpilHIJK突变体形成了规则排列且大小均一的独特的微菌落。讨论了有关铜绿假单胞菌生物膜发展的当前模型的结果。

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