Abstract

Quorum sensing (QS) is a bacterial cell-to-cell signaling mechanism that collectively regulates and synchronizes behaviors by means of small diffusible chemical molecules. In rhizobia, QS systems usually relies on the synthesis and detection of N-acyl-homoserine lactones (AHLs). In the model bacterium Sinorhizobium meliloti functions regulated by the QS systems TraI-TraR and SinI-SinR(-ExpR) include plasmid transfer, production of surface polysaccharides, motility, growth rate and nodulation. These systems are also present in other bacteria of the Sinorhizobium genus, with variations at the species and strain level. In Sinorhizobium fredii NGR234 phenotypes regulated by QS are plasmid transfer, growth rate, sedimentation, motility, biofilm formation, EPS production and copy number of the symbiotic plasmid (pSym). The analysis of the S. fredii HH103 genomes reveal also the presence of both QS systems. In this manuscript we characterized the QS systems of S. fredii HH103, determining that both TraI and SinI AHL-synthases proteins are responsible of the production of short- and long-chain AHLs, respectively, at very low and not physiological concentrations. Interestingly, the main HH103 luxR-type genes, expR and traR, are split into two ORFs, suggesting that in S. fredii HH103 the corresponding carboxy-terminal proteins, which contain the DNA-binding motives, may control target genes in an AHL-independent manner. The presence of a split traR gene is common in other S. fredii strains.

Highlights

  • The rhizobium–legume symbiosis is one of the best studied model systems of mutualistic interactions between bacteria and eukaryotic hosts

  • Supernatants from S. fredii HH103 cultures grown at stationary phase (OD600 1.2) were first assayed for acyl-homoserine lactones (AHLs) production in well diffusion assays by using the biosensor A. tumefaciens NT1

  • In order to concentrate 100-fold the putative AHLs present in HH103 supernatants, 5 mL of wild-type cultures grown at four different OD600 (0.3, 0.6, 0.9, and 1.2) were extracted with dichloromethane, evaporated, resuspended in 5 μL of methanol and analyzed by thin layer chromatography (TLC) using the same biosensor strain

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Summary

Introduction

The rhizobium–legume symbiosis is one of the best studied model systems of mutualistic interactions between bacteria and eukaryotic hosts. This symbiosis is initiated by a complex and evolved molecular exchange between both symbionts that culminate in the formation of nitrogen-fixing plant root nodules [1,2,3]. Nodulation ability of the three most studied S. fredii strains, NGR234, USDA257 and HH103 [12,13,14], is explained in part because plant flavonoids, in addition to NFs, regulate additional bacterial symbiotic-traits: Secretion of proteins through a type 3 secretion system (T3SS), exopolysaccharide (EPS) production, formation of biofilms and functioning of quorum sensing (QS) systems [15,16,17]

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