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VirSR-VirAB 系统在单核细胞增生李斯特氏菌 EGD-e 生物膜形成中的作用。

Role of the VirSR-VirAB system in biofilm formation of Listeria monocytogenes EGD-e.

机构信息

Henan Engineering Laboratory for Bioconversion Technology of Functional Microbes, College of Life Sciences, Henan Normal University, Xinxiang, China.

College of Life Sciences and Basic Medicine, Xinxiang University, Xinxiang, China.

出版信息

Food Res Int. 2021 Jul;145:110394. doi: 10.1016/j.foodres.2021.110394. Epub 2021 May 8.

Abstract

The ability of Listeria monocytogenes, an important foodborne pathogen, to form biofilms in food processing environments leads to increased opportunity for contamination of food products, which is a major concern for food safety. In this study, the role of a complex system composed of the VirSR two-component signal transduction system (TCS) and the ATP-binding cassette (ABC) transporter VirAB in biofilm formation of L. monocytogenes EGD-e was investigated. Biofilm formation was measured using the microplate assay with crystal violet staining, confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), and attachment and swarming motility were compared between strain EGD-e and its isogenic deletion mutants. Additionally, the relative expression levels of genes associated with the early steps of biofilm development in the wild-type and mutant strains were also determined by RT-qPCR. Results from microplate assay, CLSM and SEM showed that VirR is not required for biofilm formation in L. monocytogenes EGD-e. A central finding of this study is that both VirAB and VirS are essential for biofilm formation and they could function as a whole in biofilm formation of L. monocytogenes EGD-e. The results also demonstrated that both VirAB and VirS are involved in attachment, but they are not associated with swarming motility. Results from RT-qPCR showed that flaA, motA and motB were downregulated in the mutant strains ΔvirAB and ΔvirS, which could be the possible reason for reduced attachment and biofilm formation in these mutants. This study provides a better understanding of the mechanisms involved in biofilm formation of L. monocytogenes, leading to improved processes to control this biofilm-forming foodborne pathogen.

摘要

李斯特菌(Listeria monocytogenes)是一种重要的食源性致病菌,其在食品加工环境中形成生物膜的能力增加了食品产品污染的机会,这是食品安全的主要关注点。在本研究中,研究了由 VirSR 双组分信号转导系统(TCS)和 ATP 结合盒(ABC)转运蛋白 VirAB 组成的复杂系统在李斯特菌 EGD-e 生物膜形成中的作用。通过结晶紫染色的微孔板测定法、共聚焦激光扫描显微镜(CLSM)和扫描电子显微镜(SEM)测量生物膜形成,比较了菌株 EGD-e 及其同源缺失突变体之间的附着和群集运动。此外,还通过 RT-qPCR 确定了野生型和突变菌株中与生物膜发育早期步骤相关的基因的相对表达水平。微孔板测定法、CLSM 和 SEM 的结果表明,VirR 不是李斯特菌 EGD-e 生物膜形成所必需的。本研究的一个重要发现是,VirAB 和 VirS 对于李斯特菌 EGD-e 的生物膜形成都是必不可少的,它们可以作为一个整体在李斯特菌 EGD-e 的生物膜形成中发挥作用。结果还表明,VirAB 和 VirS 都参与了附着,但它们与群集运动无关。RT-qPCR 的结果表明,突变株 ΔvirAB 和 ΔvirS 中的 flaA、motA 和 motB 下调,这可能是这些突变体附着和生物膜形成减少的可能原因。本研究更好地理解了李斯特菌生物膜形成的机制,从而改进了控制这种形成生物膜的食源性致病菌的过程。

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