Calcium transcriptionally regulates movement, recombination and other functions of Xylella fastidiosa under constant flow inside microfluidic chambers.,Microbial Biotechnology

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Calcium transcriptionally regulates movement, recombination and other functions of Xylella fastidiosa under constant flow inside microfluidic chambers.
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2019-11-14 , DOI: 10.1111/1751-7915.13512
Hongyu Chen ₁ , Leonardo De La Fuente ₁

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Xylella fastidiosa is a xylem‐limited bacterial pathogen causing devastating diseases in many economically important crops. Calcium (Ca) is a major inorganic nutrient in xylem sap that influences virulence‐related traits of this pathogen, including biofilm formation and twitching motility. This study aimed to adapt a microfluidic system, which mimics the natural habitat of X. fastidiosa, for whole transcriptome analysis under flow conditions. A microfluidic chamber with two parallel channels was used, and RNA isolated from cells grown inside the system was analysed by RNA‐Seq. Ca transcriptionally regulated the machinery of type IV pili and other genes related to pathogenicity and host adaptation. Results were compared to our previous RNA‐Seq study in biofilm cells in batch cultures (Parker et al., 2016, Environ Microbiol 18, 1620). Ca‐regulated genes in both studies belonged to similar functional categories, but the number and tendencies (up‐/downregulation) of regulated genes were different. Recombination‐related genes were upregulated by Ca, and we proved experimentally that 2 mM Ca enhances natural transformation frequency. Taken together, our results suggest that the regulatory role of Ca in X. fastidiosa acts differently during growth in flow or batch conditions, and this can correlate to the different phases of growth (planktonic and biofilm) during the infection process.

中文翻译：

钙在微流室内恒定流动下转录调节小木糖的运动，重组和其他功能。

固定木糖杆菌（Xylella fastidiosa）是一种受木质部限制的细菌病原体，在许多重要的经济作物中引起毁灭性疾病。钙（Ca）是木质部树液中的主要无机养分，会影响该病原体的毒力相关特征，包括生物膜形成和抽搐。这项研究旨在适应微流控系统，该系统模仿X. fastidiosa的自然栖息地，以便在流动条件下进行整个转录组分析。使用具有两个平行通道的微流控室，并通过RNA‐Seq分析从系统内部生长的细胞中分离出的RNA。Ca转录调节IV型菌毛和其他与致病性和宿主适应性相关的基因的机制。将结果与我们先前在分批培养的生物膜细胞中进行RNA-Seq研究的结果进行了比较（Parker等。，2016，环境微生物学18， 1620）。两项研究中的钙调节基因属于相似的功能类别，但调节基因的数量和趋势（上/下调）不同。重组相关的基因被钙上调，我们通过实验证明2 mM的钙提高了自然转化的频率。两者合计，我们的结果表明，钙在X. fastidiosa中的调节作用在流量或分批条件下的生长过程中具有不同的作用，并且这可能与感染过程中不同的生长阶段（浮游生物和生物膜）相关。

更新日期：2019-11-14

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