Sinorhizobium fredii HH103 cgs Mutants Are Unable to Nodulate Determinate- and Indeterminate Nodule–Forming Legumes and Overproduce an Altered EPS

Abstract

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Sinorhizobium fredii HH103 does not strictly require KPS and/or EPS to nodulate Glycyrrhiza uralensis, an indeterminate nodule-forming legume

“…Sinorhizobium fredii HH103 produces cyclic glucans (CG) with sn-1-phosphoglycerol substituents (Crespo-Rivas et al 2009). In spite of the homology of the S. fredii rkpI gene with glycerophosphotransferases, this gene is not involved in the addition of phosphoglycerol substituents to CG, since 1 H NMR of samples from mutant SVQ538 showed the presence of CGs containing phosphoglycerol substituents (Fig.…”

“…The construction of S. fredii HH103 rkpA and rkpI mutants was carried out by using plasmid pK18mob (Schafer et al 1994) following protocols previously described (Crespo-Rivas et al 2009). Briefly, a 3 kb EcoRI fragment of cosmid pMUS766 containing the 3 0 end of the HH103 rkpA gene was subcloned into pK18mob, a suicide vector in rhizobia.…”

“…For detection of KPS, crude extracts from S. fredii mutants SVQ536 (rkpA), SVQ536 carrying cosmid pMUS766, SVQ538 (rkpI), and SVQ538 carrying cosmid pMUS759 were obtained and analyzed by NMR as previously described (Crespo-Rivas et al 2009;Hidalgo et al 2010). For KPS visualization, samples were prepared as follows: bacterial cultures were grown on liquid TY medium for 48 h. Bacterial cells were washed 3 times in 0.9% NaCl and pelleted by centrifugation.…”

“…In addition to nod genes and those directly involved in nitrogen fixation (fix and nif genes), other bacterial determinants are required for the formation of nitrogen-fixing nodules. Bacterial polysaccharides, such as cyclic glucans (CG), acidic exopolysaccharides (EPS), lipopolysaccharides (LPS), and capsular polysaccharides (KPS), are clearly among those determinants that are relevant for the establishment of an effective symbiosis (Becker and Pühler 1998;Becker et al 2005;Crespo-Rivas et al 2009;Downie 2010;Fraysse et al 2003;Kannenberg et al 1998). Rhizobial outer nodulation proteins (Nops) and different housekeeping metabolic pathways also appear important in determining the successful outcome of the symbiotic interaction Crespo-Rivas et al 2007;De Lyra et al 2006).…”

Symbiotic properties and first analyses of the genomic sequence of the fast growing model strain Sinorhizobium fredii HH103 nodulating soybean

“…2A shows the S. fredii HH103 KPS (lane 2) and LPS (lane 3) electrophoretic profiles in PAGE experiments as described by Parada et al (2006). The chemical structures of the KPS and cyclic glucans (CG) produced by S. fredii HH103 have been determined (Crespo-Rivas et al, 2009;Gil-Serrano et al, 1999).…”