Riemerella anatipestifer lacks luxS, but can uptake exogenous autoinducer-2 to regulate biofilm formation

Abstract

Riemerella anatipestifer (RA) causes major economic losses to the duck industry. Autoinducer-2 (AI-2) is a quorum-sensing signal that regulates bacterial physiology. The luxS and pfs genes are required for AI-2 synthesis in many bacterial species. pfs encodes Pfs, which functions upstream of LuxS in the biosynthesis of AI-2. In this study, we investigated the AI-2 activity of RA using an AI-2 bioassay, which showed that RA does not produce AI-2. Bioinformatic analysis indicated that the RA genome has a pfs, but not a luxS, homologue. To investigate the function of RA pfs, an avian pathogenic Escherichia coli (APEC) pfs mutant was constructed, which was subsequently transformed with a recombinant plasmid carrying RA pfs. An AI-2 bioassay demonstrated that RA pfs restored AI-2 production to the APEC pfs mutant, suggesting that RA pfs functions in AI-2 synthesis. Furthermore, we found that RA utilizes exogenous AI-2 to regulate biofilm formation. RA biofilm formation decreased significantly upon addition of exogenous AI-2. Real-time quantitative PCR results showed that expression of 13 genes related to RA biofilm formation decreased significantly when exogenous AI-2 was added to the RA culture media. These findings will benefit future studies on AI-2 regulation in RA.