Five strains of bacteria isolated from nodules of and in Yunnan Province of China were classified within the genus in the class . The highest degree of 16S rRNA gene sequence similarity was determined to be to LMG 6125 (99.7 %) and UPM-Ca7 (99.7 %). Polyphasic taxonomic methods including SDS-PAGE of whole-cell soluble proteins, comparative housekeeping sequence analysis of , and , fatty acid profiles and a series of phenotypic and physiological tests allowed us to cluster the five strains into a coherent group while differentiating them from all previously established species. The DNA–DNA relatedness between the representative strain CCBAU 65327 and the type strains of and was 26.5 and 23.4 %, respectively, clearly indicating that strain CCBAU 65327 represents a novel species for which we propose the name sp. nov. Strain CCBAU 65327 (=LMG 24762 =HAMBI 3050) is designated as the type strain, and could nodulate , , , , and in cross-nodulation tests.


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  1. Barrow, G. I. & Feltham, R. K. A.(2003).Cowan and Steel's Manual for the Identification of Medical Bacteria, 3rd edn. Cambridge: Cambridge University Press.
  2. Chen, W. X., Wang, E. T., Wang, S. Y., Li, Y. B., Chen, X. Q. & Li, Y.(1995). Characteristics of Rhizobium tianshanense sp. nov., a moderately and slowly growing root nodule bacterium isolated from an arid saline environment in Xinjiang, People's Republic of China. Int J Syst Bacteriol 45, 153–159.[CrossRef] [Google Scholar]
  3. Chen, W. F., Guan, S. H., Zhao, C. T., Yan, X. R., Man, C. X., Wang, E. T. & Chen, W. X.(2008). Different Mesorhizobium species associated with Caragana carry similar symbiotic genes and have common host ranges. FEMS Microbiol Lett 283, 203–209.[CrossRef] [Google Scholar]
  4. de Lajudie, P., Willems, A., Nick, G., Moreira, F., Molouba, F., Hoste, B., Torck, U., Neyra, M., Collins, M. D. & other authors(1998). Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp. nov. Int J Syst Bacteriol 48, 369–382.[CrossRef] [Google Scholar]
  5. De Ley, J.(1970). Reexamination of the association between melting point, buoyant density, and chemical base composition of deoxyribonucleic acid. J Bacteriol 101, 738–754. [Google Scholar]
  6. 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]
  7. Fan, S. Y., Freedman, B. & Gao, J. X.(2007). Potential environmental benefits from increased use of bioenergy in China. Environ Manage 40, 504–515.[CrossRef] [Google Scholar]
  8. Felsenstein, J.(1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783–791.[CrossRef] [Google Scholar]
  9. 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]
  10. Gao, J. L., Turner, S. L., Kan, F. L., Wang, E. T., Tan, Z. Y., Qiu, Y. H., Gu, J., Terefework, Z., Young, J. P. & other authors(2004).Mesorhizobium septentrionale sp. nov. and Mesorhizobium temperatum sp. nov., isolated from Astragalus adsurgens growing in the northern regions of China. Int J Syst Evol Microbiol 54, 2003–2012.[CrossRef] [Google Scholar]
  11. 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]
  12. Graham, P. H., Sadowsky, M. J., Keyser, H. H., Barnet, Y. M., Bradley, R. S., Cooper, J. E., De Ley, D. 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]
  13. Guan, S. H., Chen, W. F., Wang, E. T., Lu, Y. L., Yan, X. R., Zhang, X. X. & Chen, W. X.(2008).Mesorhizobium caraganae sp. nov., a novel rhizobial species nodulated with Caragana spp. in China. Int J Syst Evol Microbiol 58, 2646–2653.[CrossRef] [Google Scholar]
  14. Han, T. X., Han, L. L., Wu, L. J., Chen, W. F., Sui, X. H., Gu, J. G., Wang, E. T. & Chen, W. X.(2008).Mesorhizobium gobiense sp. nov. and Mesorhizobium tarimense sp. nov., isolated from wild legumes growing in desert soils of Xinjiang, China. Int J Syst Evol Microbiol 58, 2610–2618.[CrossRef] [Google Scholar]
  15. Haukka, K., Lindstrom, K. & Young, J. P. W.(1998). Three phylogenetic groups of nodA and nifH genes in Sinorhizobium and Mesorhizobium isolates from leguminous trees growing in Africa and Latin America. Appl Environ Microbiol 64, 419–426. [Google Scholar]
  16. Jarvis, B. D. W., Van Berkum, P., Chen, W. X., Nour, S. M., Fernandez, M. P., Cleyet-Marel, J. C. & Gillis, M.(1997). Transfer of Rhizobium loti, Rhizobium huakuii, Rhizobium ciceri, Rhizobium mediterraneum, and Rhizobium tianshanense to Mesorhizobium gen. nov. Int J Syst Bacteriol 47, 895–898.[CrossRef] [Google Scholar]
  17. Jukes, T. H. & Cantor, C. R.(1969). Evolution of protein molecules. In Mammalian Protein Metabolism, vol. 3, pp. 21–132. Edited by H. N. Munro. New York: Academic Press.
  18. Laguerre, G., Nour, S. M., Macheret, V., Sanjuan, J., Drouin, P. & Amarger, N.(2001). Classification of rhizobia based on nodC and nifH gene analysis reveals a close phylogenetic relationship among Phaseolus vulgaris symbionts. Microbiology 147, 981–993. [Google Scholar]
  19. Liu, J., Wang, E. T. & Chen, W. X.(2005). Diverse rhizobia associated with woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the temperate zone of China. Syst Appl Microbiol 28, 465–477.[CrossRef] [Google Scholar]
  20. Lu, Y. L., Chen, W. F., Wang, E. T., Guan, S. H., Yan, X. R. & Chen, W. X.(2009). Genetic diversity and biogeography of rhizobia associated with Caragana species in three ecological regions of China. Syst Appl Microbiol 32, 351–361.[CrossRef] [Google Scholar]
  21. Marmur, J.(1961). A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3, 208–218.[CrossRef] [Google Scholar]
  22. Nour, S. M., Fernandez, M. P., Normand, P. & Cleyet-Marel, J. C.(1994).Rhizobium ciceri sp. nov., consisting of strains that nodulate chickpeas (Cicer arietinum L.). Int J Syst Bacteriol 44, 511–522.[CrossRef] [Google Scholar]
  23. Saitou, N. & Nei, M.(1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425. [Google Scholar]
  24. Sasser, M.(1990).Identification of bacteria by gas chromatography of cellular fatty acids, MIDI Technical Note 101. Newark, DE: MIDI Inc.
  25. Schutter, M. E. & Dick, R. P.(2000). Comparison of fatty acid methyl ester (fame) methods for characterizing microbial communities. Soil Sci Soc Am J 64, 1659–1668.[CrossRef] [Google Scholar]
  26. Smibert, R. M. & Krieg, N. R.(1994). Phenotypic characterization. In Methods for General and Molecular Bacteriology, pp. 607–654. Edited by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg. Washington, DC: American Society for Microbiology.
  27. Su, Y. Z., Zhang, T. H., Li, Y. L. & Wang, F.(2005). Changes in soil properties after establishment of Artemisia halodendron and Caragana microphylla on shifting sand dunes in semiarid Horqin sandy land, northern China. Environ Manage 36, 272–281.[CrossRef] [Google Scholar]
  28. Tamura, K., Dudley, J., Nei, M. & Kumar, S.(2007).mega4: molecular evolutionary genetics analysis (mega) software version 4.0. Mol Biol Evol 24, 1596–1599.[CrossRef] [Google Scholar]
  29. Tan, Z. Y., Xu, X. D., Wang, E. T., Gao, J. L., Martinez-Romero, E. & Chen, W. X.(1997). Phylogenetic and genetic relationships of Mesorhizobium tianshanense and related rhizobia. Int J Syst Bacteriol 47, 874–879.[CrossRef] [Google Scholar]
  30. Tighe, S. W., de Lajudie, P., Dipietro, K., Lindstrom, 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]
  31. Turner, S. L. & Young, J. P. W.(2000). The glutamine synthetases of rhizobia: phylogenetics and evolutionary implications. Mol Biol Evol 17, 309–319.[CrossRef] [Google Scholar]
  32. Ueda, T., Suga, Y., Yahiro, N. & Matsuguchi, T.(1995). Phylogeny of Sym plasmids of rhizobia by PCR-based sequencing of a nodC segment. J Bacteriol 177, 468–472. [Google Scholar]
  33. Vauterin, L. & Vauterin, P.(1992). Computer-aided objective comparison of electrophoresis patterns for grouping and identification of microorganisms. Eur Microbiol 1, 37–41. [Google Scholar]
  34. Velazquez, E., Igual, J. M., Willems, A., Fernández, M. P., Muñoz, E., Mateos, P. F., Abril, A., Toro, N., Normand, P. & other authors(2001).Mesorhizobium chacoense sp. nov., a novel species that nodulates Prosopis alba in the Chaco Arido region (Argentina). Int J Syst Evol Microbiol 51, 1011–1021.[CrossRef] [Google Scholar]
  35. Vincent, J. M.(1970). The cultivation, isolation and maintenance of rhizobia. In A Manual for the Practical Study of the Root-Nodule Bacteria, pp. 1–13. Edited by J. M. Vincent. Oxford: Blackwell Scientific.
  36. Wang, E. T., van Berkum, P., Sui, X. H., Beyene, D., Chen, W. X. & Martinez-Romero, E.(1999). Diversity of rhizobia associated with Amorpha fruticosa isolated from Chinese soils and description of Mesorhizobium amorphae sp. nov. Int J Syst Bacteriol 49, 51–65.[CrossRef] [Google Scholar]
  37. Wang, F. Q., Wang, E. T., Liu, J., Chen, Q., Sui, X. H., Chen, W. F. & Chen, W. X.(2007).Mesorhizobium albiziae sp. nov., a novel bacterium that nodulates Albizia kalkora in a subtropical region of China. Int J Syst Evol Microbiol 57, 1192–1199.[CrossRef] [Google Scholar]
  38. Wayne, L. G., Brenner, D. J., Colwell, R. R., Grimont, P. A. D., Kandler, O., Krichevsky, M. I., Moore, L. H., Moore, W. E. C., Murray, R. G. E. & other authors(1987). International Committee on Systematic Bacteriology. Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int J Syst Bacteriol 37, 463–464.[CrossRef] [Google Scholar]
  39. Yan, X. R., Chen, W. F., Fu, J. F., Lu, Y. L., Xue, C. Y., Sui, X. H., Li, Y., Wang, E. T. & Chen, W. X.(2007).Mesorhizobium spp. are the main microsymbionts of Caragana spp. grown in Liaoning Province of China. FEMS Microbiol Lett 271, 265–273.[CrossRef] [Google Scholar]
  40. Zhang, M. L.(2005). A dispersal and vicariance analysis of the genus Caragana Fabr. J Integr Plant Biol 47, 897–904.[CrossRef] [Google Scholar]

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vol. , part 12, pp. 3012 - 3018

[ PDF] (195KB), containing:

Dendrogram based upon SDS-PAGE of whole-cell soluble proteins.

Comparison of partial (a) and (b) gene phylogenies, showing that the novel group has close relationships with the type strains of , , and and is distantly related to the other species of the genus .

Sequence similarities of 16S rRNA gene, , , , and and DNA relatedness among novel strain CCBAU 65327 and related strains.

Cellular fatty acid compositions of strain CCBAU 65327 and previously reported species of the genus .


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