Six strains were isolated from root nodules of grown in Nanchang, Yifeng, Taihu, Huaibei, Bengbu and Lujiang, in the middle and lower reaches of the Yangtze River. According to phylogenetic analyses of 16S rRNA gene, and sequences, these strains belong to the genus , with and as the closest related species. CCBAU 33202, a representative of these novel isolates, showed sequence similarity to its closest relatives CFN 42 and USDA 2370 of 99.5 and 99.1 % for the 16S rRNA gene, 91.9 and 91.9 % for and 90.3 and 93.2 % for . The strains from this study could also be differentiated from CFN 42 and USDA 2370 by 16S–IGS RFLP and SDS-PAGE of whole-cell proteins, fatty acid profiles and several phenotypic characteristics. DNA–DNA hybridization yielded relatedness of 19 and 14–43 %, respectively, with CFN 42 and strains representing different biovars of . All data obtained in this study showed that these isolates belong to a novel species, for which the name sp. nov. is proposed. The type strain, CCBAU 33202 (=LMG 23997 =JCM 14381), was isolated from Nanchang.


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  1. Amarger, N., Macheret, V. & Laguerre, G.(1997).Rhizobium gallicum sp. nov. and Rhizobium giardinii sp. nov., from Phaseolus vulgaris nodules. Int J Syst Bacteriol 47, 996–1006.[CrossRef] [Google Scholar]
  2. Chen, W. X., Tan, Z. Y., Gao, J. L., Li, Y. & Wang, E. T.(1997).Rhizobium hainanense sp. nov., isolated from tropical legumes. Int J Syst Bacteriol 47, 870–873.[CrossRef] [Google Scholar]
  3. De Ley, J., Cattoir, H. & Reynaerts, A.(1970). The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem 12, 133–142.[CrossRef] [Google Scholar]
  4. Gao, J. L., Sun, J. G., Li, Y., Wang, E. T. & Chen, W. X.(1994). Numerical taxonomy and DNA relatedness of tropical rhizobia isolated from Hainan Province, China. Int J Syst Bacteriol 44, 151–158.[CrossRef] [Google Scholar]
  5. Gaunt, M. W., Turner, S. L., Rigottier-Gois, L., Lloyd-Macgilp, S. A. & Young, J. P. W.(2001). Phylogenies of atpD and recA support the small subunit rRNA-based classification of rhizobia. Int J Syst Evol Microbiol 51, 2037–2048.[CrossRef] [Google Scholar]
  6. Graham, P. H., Sadowsky, M. J., Keyser, H. H., Barnet, Y. M., Bradley, R. S., Cooper, J. E., De Ley, J., Jarvis, B. D. W., Roslycky, E. B. & other authors(1991). Proposed minimal standards for the description of new genera and species of root- and stem-nodulating bacteria. Int J Syst Bacteriol 41, 582–587.[CrossRef] [Google Scholar]
  7. Graham, P. H., Sadowsky, M. J., Tighe, S. W., Thompson, J. A., Date, R. A., Howieson, J. G. & Thomas, R.(1995). Differences among strains of Bradyrhizobium in fatty acid-methyl ester analysis. Can J Microbiol 41, 1038–1042.[CrossRef] [Google Scholar]
  8. Hall, T. A.(1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41, 95–98. [Google Scholar]
  9. Jarvis, B. D. W. & Tighe, S. W.(1994). Rapid identification of Rhizobium species based on cellular fatty acid analysis. Plant Soil 161, 31–41.[CrossRef] [Google Scholar]
  10. Jeanmougin, F., Thompson, J. D., Gouy, M., Higgins, D. G. & Gibson, T. J.(1998). Multiple sequence alignment with clustal_x. Trends Biochem Sci 23, 403–405.[CrossRef] [Google Scholar]
  11. Jordan, D. C.(1984). Genus I. Rhizobium Frank 1889, 338AL. In Bergey's Manual of Systematic Bacteriology, vol. 1, pp. 235–242. Edited by N. R. Krieg & J. G. Holt. Baltimore: Williams & Wilkins.
  12. Kumar, S., Tamura, K. & Nei, M.(2004).mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef] [Google Scholar]
  13. Laguerre, G., Louvrier, P., Allard, M. R. & Amarger, N.(2003). Compatibility of rhizobial genotypes within natural populations of Rhizobium leguminosarum biovar viciae for nodulation of host legumes. Appl Environ Microbiol 69, 2276–2283.[CrossRef] [Google Scholar]
  14. Mutch, L. A. & Young, J. P. W.(2004). Diversity and specificity of Rhizobium leguminosarum biovar viciae on wild and cultivated legumes. Mol Ecol 13, 2435–2444.[CrossRef] [Google Scholar]
  15. Mutch, L. A., Tamimi, S. M. & Young, J. P. W.(2003). Genotypic characterisation of rhizobia nodulating Vicia faba from the soils of Jordan: a comparison with UK isolates. Soil Biol Biochem 35, 709–714.[CrossRef] [Google Scholar]
  16. Nichols, R.(2001). Gene trees and species trees are not the same. Trends Ecol Evol 16, 358–364.[CrossRef] [Google Scholar]
  17. Rasolomampianina, R., Bailly, X., Fetiarison, R., Rabevohitra, R., Béna, G., Ramaroson, L., Raherimandimby, M., Moulin, L., De Lajudie, P. & other authors(2005). Nitrogen-fixing nodules from rose wood legume trees (Dalbergia spp.) endemic to Madagascar host seven different genera belonging to α- and β-Proteobacteria. Mol Ecol 14, 4135–4146.[CrossRef] [Google Scholar]
  18. Rosenberg, N. A.(2002). The probability of topological concordance of gene trees and species trees. Theor Popul Biol 61, 225–247.[CrossRef] [Google Scholar]
  19. Stackebrandt, E., Frederiksen, W., Garrity, G. M., Grimont, P. A., Kämpfer, P., Maiden, M. C., Nesme, X., Rosselló-Mora, R., Swings, J. & other authors(2002). Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology. Int J Syst Evol Microbiol 52, 1043–1047.[CrossRef] [Google Scholar]
  20. Tan, Z. Y., Wang, E. T., Gao, J. L., Martinez-Romero, E. & Chen, W. X.(1997). Phylogenetic and genetic relationships of Mesorhizobium tianshanese and related rhizobia. Int J Syst Bacteriol 47, 874–879.[CrossRef] [Google Scholar]
  21. Terefework, Z., Kaijalainen, S. & Lindström, K.(2001). AFLP fingerprinting as a tool to study the genetic diversity of Rhizobium galegae isolated from Galega orientalis and Galega officinalis. J Biotechnol 91, 169–180.[CrossRef] [Google Scholar]
  22. Thompson, J. D., Higgins, D. G. & Gibson, T. J.(1994).clustalw: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673–4680.[CrossRef] [Google Scholar]
  23. Tian, C. F., Wang, E. T., Han, T. X., Sui, X. H. & Chen, W. X.(2007). Genetic diversity of rhizobia associated with Vicia faba in three ecological regions of China. Arch Microbiol 188, 273–282.[CrossRef] [Google Scholar]
  24. Tighe, S. W., de Lajudie, P., Dipietro, K., Lindström, K., Nick, G. & Jarvis, B. D. W.(2000). Analysis of cellular fatty acids and phenotypic relationships of Agrobacterium, Bradyrhizobium, Mesorhizobium, Rhizobium and Sinorhizobium species using the Sherlock Microbial Identification System. Int J Syst Evol Microbiol 50, 787–801.[CrossRef] [Google Scholar]
  25. Valverde, A., Igual, J. M., Peix, A., Cervantes, E. & Velázquez, E.(2006).Rhizobium lusitanum sp. nov., a bacterium that nodulates Phaseolus vulgaris. Int J Syst Evol Microbiol 56, 2631–2637.[CrossRef] [Google Scholar]
  26. van Berkum, P., Beyene, D., Vera, F. T. & Keyser, H. H.(1995). Variability among Rhizobium strains originating from nodules of Vicia faba. Appl Environ Microbiol 61, 2649–2653. [Google Scholar]
  27. Vincent, J. M.(1970).A Manual for the Practical Study of the Root-nodule Bacteria. Oxford: Blackwell Scientific.
  28. Vinuesa, P., Silva, C., Werner, D. & Martinez-Romero, E.(2005). Population genetics and phylogenetic inference in bacterial molecular systematics: the roles of migration and recombination in Bradyrhizobium species cohesion and delineation. Mol Phylogenet Evol 34, 29–54.[CrossRef] [Google Scholar]
  29. Wei, G. H., Tan, Z. Y., Zhu, M. E., Wang, E. T., Han, S. Z. & Chen, W. X.(2003). Characterization of rhizobia isolated from legume species within the genera Astragalus and Lespedeza grown in the Loess Plateau of China and description of Rhizobium loessense sp. nov. Int J Syst Evol Microbiol 53, 1575–1583.[CrossRef] [Google Scholar]
  30. Young, J. P. W., Mutch, L. A., Ashford, D. A., Zézé, A. & Mutch, K. E.(2003). The molecular evolution of host specificity in the rhizobium-legume symbiosis. In Genes in the Environment, pp. 245–257. Edited by R. Hails, H. C. J. Godfray & J. Beringer. Oxford: Blackwell Science.

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vol. , part 12, pp. 2871 - 2875

Neighbour-joining trees based on partial (Fig. S1) and (Fig. S2) gene sequences of strains of sp. nov. and related species.

Dendrograms of 16S–IGS RFLP profiles (Fig. S3) and whole-cell protein SDS-PAGE profiles (Fig. S4) showing the relationship of the novel isolates ( sp. nov.) and related species.

Neighbour-joining tree based on sequences.

DNA–DNA hybridization of sp. nov. CCBAU 33202 with related strains.

[PDF file of Supplementary Figures and Table](228 KB)

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