The role and molecular mechanism of flgK gene in biological properties, pathogenicity and virulence genes expression of Aeromonas hydrophila

Aeromonas hydrophila is a highly pathogenic aquatic resident bacterium that can cause co-morbidity in aquatic animals, waterfowl, poultry, and humans. Flagellum is the motility organ of bacteria important for bacterium tissue colonization and invasion. The flgK gene encodes a flagellar hook protein essential for normal flagellar formation. In order to explore the role of flgK in A. hydrophila, a flgK gene mutant strain of A. hydrophila (∆flgK-AH) was constructed using an efficient suicide plasmid-mediated homologous recombination method, and gene sequencing confirmed successful mutation of the flgK gene. The biological properties, pathogenicity and virulence genes expression were compared. The results showed that there was no significant difference in the growth, hemolytic, and swarming abilities, but the swimming and biofilm formation abilities of ∆flgK-AH were significantly reduced and the transmission electron microscope (TEM) results showed that the ∆flgK-AH strain did not have a flagellar structure. The median lethal dose (LD50) value of the ∆flgK-AH in Carassius auratus was 1.47-fold higher than that of the wild-type strain (WT-AH). The quantitative real-time PCR results showed that only the expression level of the lapA gene was up-regulated by 1.47 times compared with the WT-AH, while the expression levels of other genes were significantly down-regulated. In conclusion, flgK gene mutant led to a decline in the pathogenicity possibly by reducing swimming and biofilm formation abilities, these biological properties might result from the down-regulated expression of flagellate and pilus-related genes.