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Transcriptome analysis of early surface-associated growth of Shewanella oneidensis MR-1.

机译:转录组学分析沙伊氏假单胞菌MR-1的早期表面相关生长。

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摘要

Bacterial biofilm formation starts with single cells attaching to a surface, however, little is known about the initial attachment steps and the adaptation to the surface-associated life style. Here, we describe a hydrodynamic system that allows easy harvest of cells at very early biofilm stages. Using the metal ion-reducing gammaproteobacterium Shewanella oneidensis MR-1 as a model organism, we analyzed the transcriptional changes occurring during surface-associated growth between 15 and 60 minutes after attachment. 230 genes were significantly upregulated and 333 were downregulated by a factor of ≥ 2. Main functional categories of the corresponding gene products comprise metabolism, uptake and transport, regulation, and hypothetical proteins. Among the genes highly upregulated those implicated in iron uptake are highly overrepresented, strongly indicating that S. oneidensis MR-1 has a high demand for iron during surface attachment and initial biofilm stages. Subsequent microscopic analysis of biofilm formation under hydrodynamic conditions revealed that addition of Fe(II) significantly stimulated biofilm formation of S. oneidensis MR-1 while planktonic growth was not affected. Our approach to harvest cells for transcriptional analysis of early biofilm stages is expected to be easily adapted to other bacterial species.
机译:细菌生物膜的形成始于单个细胞附着在表面上,但是,关于初始附着步骤以及对与表面相关的生活方式的适应知之甚少。在这里,我们描述了一种流体动力学系统,该系统可以在非常早期的生物膜阶段轻松收获细胞。使用减少金属离子的伽玛变形杆菌Shewanella oneidensis MR-1作为模型生物,我们分析了附着后15至60分钟之间表面相关生长过程中发生的转录变化。 ≥2的因子使230个基因显着上调,而333个基因下调。相应基因产物的主要功能类别包括代谢,摄取和转运,调节和假设的蛋白质。在高度上调的基因中,与铁吸收有关的那些基因高度被过度表达,这强烈表明在表面附着和初始生物膜阶段,沙门氏菌MR-1对铁的需求很高。随后在水动力条件下对生物膜形成的显微​​分析表明,添加Fe(II)可以显着刺激沙棘线虫MR-1的生物膜形成,而浮游生物的生长则不受影响。预期我们的用于早期生物膜阶段转录分析的收获细胞方法很容易适应其他细菌物种。

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