This study firstly demonstrated that H. pylori infection contributes to IRX1 promoter methylation on gastric mucosa.
Four rhizobial strains, designated CCBAU 85046 T , CCBAU 85051, CCBAU 85048 and CCBAU 85049, isolated from root nodules of Oxytropis glabra grown in Tibet, China, were previously defined, using amplified 16S rRNA gene restriction analysis, as a novel group within the genus Rhizobium. To clarify their taxonomic position, these strains were further analysed and compared with reference strains of related bacteria using a polyphasic approach. The 16S rRNA gene analysis showed that the four isolates formed a distinct phylogenetic lineage in the genus Rhizobium. The isolates showed highest sequence similarity (97.8 %) to Rhizobium indigoferae CCBAU 71042 T . Phenotypic and physiological tests, DNA-DNA hybridization, phylogenetic analyses of housekeeping genes recA, atpD and glnII and fatty acid profiles also indicated that these four strains constitute a novel group distinct from recognized species of the genus Rhizobium. , CCBAU 85048, CCBAU 85049 and CCBAU 85051 as a novel group belonging to the genus Rhizobium based on restriction fragment length polymorphism analysis of amplified 16S rRNA genes. To clarify the taxonomic status of this group, in the present study the four strains were further characterized and compared with reference strains by using a polyphasic approach.The four test strains and reference strains for species of the genus Rhizobium were obtained from the corresponding culture collection centres and maintained on YMA medium (Vincent, 1970) at 4 u C for temporary storage. Genomic DNA was extracted from the strains according to the protocol of Terefework et al. (2001) and was used as a template in the amplification of different genes or specific Abbreviation: IGS, intergenic spacer.The GenBank/EMBL/DDBJ accession numbers for the partial 16S rRNA, 16S-23S rRNA intergenic spacer, atpD, recA and glnII gene sequences of strain CCBAU 85046 T are EU256434, EU288745, EU288670, EU288696 and DQ985380, respectively. The GenBank/ EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains CCBAU 85051, CCBAU 85048 and CCBAU 85049 are GU938296, GU938297 and GU938298, respectively. DNA fragments. The 16S rRNA gene of each of the four strains was amplified using primers fD1 and rD1 as described previously (Weisburg et al., 1991) and sequenced directly (van Berkum et al., 1996). The 16S rRNA gene sequences of type strains of species of the genus Rhizobium were obtained from the GenBank database. All sequences were aligned using the CLUSTAL W program in MEGA 4.0 software (Tamura et al., 2007). Phylogenetic distances were calculated according to the model of Jukes & Cantor (1969). Phylogenetic trees were reconstructed using the neighbour-joining (Saitou & Nei, 1987) and maximumlikelihood (Felsenstein, 1985) methods with bootstrap values based on 1000 replicates. The topology of the maximum-likelihood tree (not shown) was similar to that of the neighbour-joining tree (Fig. 1). In the phylogenetic tree ( Fig. 1 T , which showed sequence similarities ranging between 97.6 and 96.9 %.Since the 16S-23S intergeni...
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