Identification of Quorum-Sensing Signal Molecules and the LuxRI Homologs in Fish Pathogen Edwardsiella tarda

Abstract

Edwardsiella tarda is a gram-negative bacterium that causes septicaemia in fish and serious damage to the aquaculture industry. The virulence factors of this pathogen and control mechanisms of the expression of virulence genes have not yet been clearly elucidated. A number of gram-negative pathogenic bacteria have a quorum-sensing system. These bacteria produce N-acyl-L-homoserine lactone (AHL) that they use them as a quorum-sensing signal molecule. In this study, we found that E. tarda isolated from deceased flounder produces AHLs. Thin layer chromatography analysis indicated that the two kinds of AHL produced by E. tarda seemed to be N-hexanoyl-L-homoserine lactone (C6-HSL) and N-heptanoyl-L-homoserine lactone (C7-HSL). We have cloned and sequenced the quorum-sensing genes, luxI homolog (edwI) and luxR homolog (edwR). EdwI and EdwR showed high identity with CarI/CarR and ExpI/ExpR from Erwinia carotovora, respectively. SDS–PAGE analysis of extracellular proteins revealed that the expression of the 55-kDa protein, which was reported as a virulent-strain-specific protein, is controlled by AHLs. These results suggest that some virulence factors are regulated by the quorum-sensing system in E. tarda.