Two facultatively anaerobic, nitrogen-fixing bacteria (strains MSSRF30 and MSSRF31) were isolated from a mangrove-associated wild rice ( Tateoka). These strains were determined to be nitrogen-fixers using the acetylene reduction assay and by PCR detection of a gene amplicon. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the novel strains were most closely related to LMG 7894 (96.8 % gene sequence similarity), LMG 7910 (96.8 % sequence similarity) and CIP 102760 (96.7 % sequence similarity). Further multilocus sequence analysis using , , and genes also showed low levels of sequence similarities (83–93 %) with all species of the genus with validly published names. A multigene phylogenetic tree using concatenated sequences of the four genes (16S rRNA, , and ) showed that strains MSSRF30 and MSSRF31 occupied a distinct phylogenetic position, forming a long branching that was not clustered with any other recognized species. The fatty acid profile also suggested that the novel strains belonged to the genus . The results of physiological and biochemical tests, genomic fingerprinting and DNA–DNA hybridization analyses clearly differentiated both novel strains from their closest phylogenetic neighbours, IID6019, LMG 7896, LMG 7894 and LMG 7910. Several phenotypic traits enabled the differentiation of strain MSSRF30 from other species of the genus . The DNA G+C content of strain MSSRF30 was 44.4±3.1 mol%. Based on genotypic, phenotypic, chemotaxonomic, phylogenetic and DNA–DNA hybridization analyses, the name sp. nov. (type strain MSSRF30=LMG 24061=DSM 19223) is proposed for this novel taxon.


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