Abstract

Paraburkholderia phymatum is a rhizobial strain that belongs to the beta-proteobacteria, a group known to form efficient nitrogen-fixing symbioses within root nodules of several legumes, including the agriculturally important common bean. The establishment of the symbiosis requires the exchange of rhizobial and plant signals such as lipochitooligosaccharides (Nod factors), polysaccharides, and flavonoids. Inspection of the genome of the competitive rhizobium P. phymatum revealed the presence of several polysaccharide biosynthetic gene clusters. In this study, we demonstrate that bceN, a gene encoding a GDP-D-mannose 4,6-dehydratase, which is involved in the production of the exopolysaccharide cepacian, an important component of biofilms produced by closely related opportunistic pathogens of the Burkholderia cepacia complex (Bcc), is required for efficient plant colonization. Wild-type P. phymatum was shown to produce cepacian while a bceN mutant did not. Additionally, the bceN mutant produced a significantly lower amount of biofilm and formed less root nodules compared to the wild-type strain with Phaseolus vulgaris as host plant. Finally, expression of the operon containing bceN was induced by the presence of germinated P. vulgaris seeds under nitrogen limiting conditions suggesting a role of this polysaccharide in the establishment of this ecologically important symbiosis.

Highlights

  • Rhizobia are phylogenetically diverse soil bacteria, which possess the ability to infect the roots of certain legumes and induce nodules on roots or stems

  • Previous work has identified the CEP biosynthetic gene cluster in P. phymatum (Ferreira et al, 2010). The products of this gene cluster containing 22 genes showed high amino acid identity with enzymes required for the biosynthesis of the polysaccharide CEP in the opportunistic pathogen Burkholderia cepacia complex (Bcc) strains B. cenocepacia H111 and B. cenocepacia J2315 (Figure 1)

  • Rhizobial surface polysaccharides represent the interface between bacteria and roots in the soil and are important traits for the establishment of a successful rhizobial symbiosis (Niehaus and Becker, 1998; Fraysse et al, 2003)

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Summary

Introduction

Rhizobia are phylogenetically diverse soil bacteria, which possess the ability to infect the roots of certain legumes and induce nodules on roots or stems. The nodules represent specialized root organs in which rhizobia fix atmospheric nitrogen (N2) into ammonia, thereby giving legumes a pronounced growth advantage in nitrogen (N) deprived soils (Masson-Boivin et al, 2009; Oldroyd et al, 2011; Udvardi and Poole, 2013). Rhizobia were thought to belong exclusively to the alpha-subclass of proteobacteria (alpha-rhizobia, isolated first in 1888 by Beijerinck). This situation changed in 2001, when two genera belonging to the betaproteobacteria (Burkholderia and Cupriavidus) were discovered to form nitrogen-fixing root nodules (beta-rhizobia) (Chen et al, 2001; Moulin et al, 2001; Bontemps et al, 2010). For the legume nodulating strains that belong to the environmental clade of the genus Burkholderia, the new genus Paraburkholderia has been recently proposed (Sawana et al, 2014; Beukes et al, 2017).

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