Vibrio cholerae O1, a natural inhabitant of estuarine environments, is found in a dormant, viable but non‐culturable (VBNC) state during interepidemic periods. Although the individual roles of abiotic factors affecting VBNC formation have been extensively studied, their interplay in driving this phenomenon remains largely unaddressed. Here, we identified that major abiotic factors synergize with low nutrient conditions governing entry of cells into the VBNC state. Specifically, V. cholerae cells exposed to a combination of alkaline pH and high salinity under aeration at low temperatures (VBNC‐inducing conditions) synergize to facilitate rapid entry into VBNC, whereas the opposite conditions prevented entry into the state. The major virulence regulator ToxR, and the stringent response protein RelA played opposing roles, repressing and facilitating VBNC entry respectively. Further, VBNC‐inducing conditions negated the effects of ToxR and RelA, facilitating rapid formation of VBNC cells. In summary, this study highlights the synergy between critical abiotic factors and identified ToxR and RelA as two associated regulators, allowing for the persistence of V. cholerae in aquatic environments. Insights obtained in this study will help better understand environmental survival non‐sporulating bacteria and transmission of facultative bacterial pathogens.

中文翻译：

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非生物因子驱动活的但不可培养的霍乱弧菌的协同作用。

霍乱弧菌O1是河口环境的自然栖息地，在流行期间，处于休眠，可生存但不可培养的状态（VBNC）。尽管已经广泛研究了影响VBNC形成的非生物因素的个体作用，但它们在驱动这种现象中的相互作用仍未得到解决。在这里，我们确定了主要的非生物因子与控制细胞进入VBNC状态的低营养条件协同作用。具体而言，霍乱弧菌在低温（VBNC诱导条件）下于曝气条件下暴露于碱性pH和高盐度的细胞协同作用，有助于快速进入VBNC，而相反的条件则阻止其进入该状态。主要的毒力调节剂ToxR和严格的应答蛋白RelA发挥相反的作用，分别抑制和促进VBNC进入。此外，VBNC诱导条件消除了ToxR和RelA的作用，促进了VBNC细胞的快速形成。总而言之，这项研究突出了关键非生物因素之间的协同作用，并确定了ToxR和RelA是两个相关的调节因子，从而使霍乱弧菌得以持久存在。在水生环境中。这项研究获得的见解将有助于更好地了解非孢子形成细菌在环境中的生存情况以及兼性细菌病原体的传播。