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Rhizobium mesosinicum sp. nov., isolated from root nodules of three different legumes

Dong Xu Lin^1,, Wen Feng Chen^1,, Feng Qin Wang^1,2, Dong Hu¹, En Tao Wang³, Xin Hua Sui¹ and Wen Xin Chen¹

¹ State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China
² College of Life Science, Henan Agricultural University, Zhengzhou 450002, PR China
³ Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, 11340 México DF, Mexico

Correspondence
Wen Xin Chen
wenxin_chen{at}263.net

Thirteen novel strains were isolated from root nodules of three different leguminous plants of the genera Albizia, Kummerowia and Dalbergia grown in China. Cells of these strains were Gram-negative, strictly aerobic, non-spore-forming, motile rods. Phylogenetic analysis of 16S rRNA gene sequences revealed that they belong to the genus Rhizobium. A representative strain, CCBAU 25010^T, showed 98.8 % 16S rRNA gene sequence similarity to its closest phylogenetic relative, Rhizobium sullae IS123^T. The 16S–23S intergenic spacer (ITS) sequence of CCBAU 25010^T shared 91.5 and 87.2 % sequence similarity, respectively, with those of Rhizobium etli CFN 42^T and Rhizobium leguminosarum LMG 14904^T. Analysis of the sequences of the housekeeping genes atpD and recA was in agreement with the results of ITS sequence analysis. The nodC gene sequence of CCBAU 25010^T was identical to that of Rhizobium tropici CFN 299. DNA–DNA hybridization values for strain CCBAU 25010^T ranged from 20.7 % (with Rhizobium mongolense USDA 1844^T) to 34.4 % (with R. leguminosarum USDA 2370^T). Cell protein SDS-PAGE, BOX-PCR and several phenotypic characteristics, such as use of sole carbon sources and antibiotic resistance, could differentiate the novel strains from defined Rhizobium species. We therefore propose that the novel strains reported in this study form a novel species, Rhizobium mesosinicum sp. nov., with the type strain CCBAU 25010^T (=LMG 24135^T =JCM 14777^T).

These authors contributed equally to this work.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene, 16S–23S ITS, atpD and recA sequences of strains CCBAU 25010^T, CCBAU 25217 and CCBAU 41044 are respectively DQ100063, EF070130 and AY395697 (16S rRNA gene), EU120729, EU120730 and DQ352862 (16S–23S ITS), EU120726, EU120727 and DQ310808 (atpD) and EU120732, EU120731 and EU034028 (recA). The accession number for the nodC sequence of strain CCBAU 25010^T is EU120728.

Neighbour-joining trees based on ITS and atpD–recA sequences, SDS-PAGE and BOX-PCR profiles of representative novel and reference strains and detailed DNA–DNA hybridization results are available as supplementary material with the online version of this paper.