Gene Analysis of Listeria monocytogenes Suspended Aggregates Induced by Ralstonia insidiosa Cell-Free Supernatants under Nutrient-Poor Environments

Listeria monocytogenes is a zoonotic food-borne pathogen. The production of food-borne pathogenic bacteria aggregates is considered to be a way to improve their resistance and persistence in the food chain. Ralstonia insidiosa has been shown to induce L. monocytogenes to form suspended aggregates, but induction mechanisms remain unclear. In the study, the effect of R. insidiosa cell-free supernatants cultured in 10% TSB medium (10% RIS) on the formation of L. monocytogenes suspended aggregates was evaluated. Next, the Illumina RNA sequencing was used to compare the transcriptional profiles of L. monocytogenes in 10% TSB medium with and without 10% RIS to identify differentially expressed genes (DEGs). The result of functional annotation analysis of DEGs indicated that these genes mainly participate in two component system, bacterial chemotaxis and flagellar assembly. Then the reaction network of L. monocytogenes suspended aggregates with the presence of 10% RIS was summarized. The gene-deletion strain of L. monocytogenes was constructed by homologous recombination. The result showed that cheA and cheY are key genes in the formation of suspended aggregates. This research is the preliminary verification of suspended aggregates’ RNA sequencing and is helpful to analyze the aggregation mechanisms of food-borne pathogenic bacteria from a new perspective.