Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Dec;63(12):4748–4758. doi: 10.1128/aem.63.12.4748-4758.1997

Genetic diversity of rhizobial symbionts isolated from legume species within the genera Astragalus, Oxytropis, and Onobrychis.

G Laguerre 1, P van Berkum 1, N Amarger 1, D Prévost 1
PMCID: PMC168797  PMID: 9406393

Abstract

The genetic diversity of 44 rhizobial isolates from Astragalus, Oxytropis, and Onobrychis spp. originating from different geographic locations was evaluated by mapped restriction site polymorphism (MRSP) analysis of 16S rRNA genes and by PCR DNA fingerprinting with repetitive sequences (REP-PCR). A comparison of tree topologies of reference strains constructed with data obtained by MRSP and by 16S rRNA gene sequence analyses showed that the topologies were in good agreement, indicating that the MSRP approach results in reasonable estimates of rhizobial phylogeny. The isolates were distributed into 14 distinct 16S rRNA gene types clustering into three major groups which corresponded with three of the genera within the legume symbionts. Most of the isolates were within the genus Mesorhizobium. Five were identified with different genomic species nodulating Lotus spp. and Cicer arietinum. Three Astragalus isolates were classified as Bradyrhizobium, one being similar to Bradyrhizobium elkanii and another being similar to Bradyrhizobium japonicum. Six of the isolates were related to species within the genus Rhizobium. Two were similar to Rhizobium leguminosarum, and the remainder were identified as Rhizobium gallicum. DNA fingerprinting by REP-PCR revealed a high level of diversity within single 16S ribosomal DNA types. The 44 isolates were distributed into 34 REP groups. Rhizobial classification at the genus and probably also the species levels was independent of geographic origin and host plant affinity.

Full Text

The Full Text of this article is available as a PDF (378.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amarger N., Macheret V., Laguerre G. Rhizobium gallicum sp. nov. and Rhizobium giardinii sp. nov., from Phaseolus vulgaris nodules. Int J Syst Bacteriol. 1997 Oct;47(4):996–1006. doi: 10.1099/00207713-47-4-996. [DOI] [PubMed] [Google Scholar]
  2. Beringer J. E. R factor transfer in Rhizobium leguminosarum. J Gen Microbiol. 1974 Sep;84(1):188–198. doi: 10.1099/00221287-84-1-188. [DOI] [PubMed] [Google Scholar]
  3. Chen W. X., Tan Z. Y., Gao J. L., Li Y., Wang E. T. Rhizobium hainanense sp. nov., isolated from tropical legumes. Int J Syst Bacteriol. 1997 Jul;47(3):870–873. doi: 10.1099/00207713-47-3-870. [DOI] [PubMed] [Google Scholar]
  4. Chen W., Wang E., Wang S., Li Y., Chen X., Li Y. 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. 1995 Jan;45(1):153–159. doi: 10.1099/00207713-45-1-153. [DOI] [PubMed] [Google Scholar]
  5. Dessen P., Fondrat C., Valencien C., Mugnier C. BISANCE: a French service for access to biomolecular sequence databases. Comput Appl Biosci. 1990 Oct;6(4):355–356. doi: 10.1093/bioinformatics/6.4.355. [DOI] [PubMed] [Google Scholar]
  6. Dupuy N., Willems A., Pot B., Dewettinck D., Vandenbruaene I., Maestrojuan G., Dreyfus B., Kersters K., Collins M. D., Gillis M. Phenotypic and genotypic characterization of bradyrhizobia nodulating the leguminous tree Acacia albida. Int J Syst Bacteriol. 1994 Jul;44(3):461–473. doi: 10.1099/00207713-44-3-461. [DOI] [PubMed] [Google Scholar]
  7. Eardly B. D., Materon L. A., Smith N. H., Johnson D. A., Rumbaugh M. D., Selander R. K. Genetic structure of natural populations of the nitrogen-fixing bacterium Rhizobium meliloti. Appl Environ Microbiol. 1990 Jan;56(1):187–194. doi: 10.1128/aem.56.1.187-194.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eardly B. D., Young J. P., Selander R. K. Phylogenetic position of Rhizobium sp. strain Or 191, a symbiont of both Medicago sativa and Phaseolus vulgaris, based on partial sequences of the 16S rRNA and nifH genes. Appl Environ Microbiol. 1992 Jun;58(6):1809–1815. doi: 10.1128/aem.58.6.1809-1815.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  10. Jensen H. L. Mutual host plant relationships in two groups of legume root nodule bacteria (Rhizobium spp.). Arch Mikrobiol. 1967;59(1):174–179. doi: 10.1007/BF00406329. [DOI] [PubMed] [Google Scholar]
  11. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980 Dec;16(2):111–120. doi: 10.1007/BF01731581. [DOI] [PubMed] [Google Scholar]
  12. Laguerre G., Allard M. R., Revoy F., Amarger N. Rapid Identification of Rhizobia by Restriction Fragment Length Polymorphism Analysis of PCR-Amplified 16S rRNA Genes. Appl Environ Microbiol. 1994 Jan;60(1):56–63. doi: 10.1128/aem.60.1.56-63.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laguerre G., Mavingui P., Allard M. R., Charnay M. P., Louvrier P., Mazurier S. I., Rigottier-Gois L., Amarger N. Typing of rhizobia by PCR DNA fingerprinting and PCR-restriction fragment length polymorphism analysis of chromosomal and symbiotic gene regions: application to Rhizobium leguminosarum and its different biovars. Appl Environ Microbiol. 1996 Jun;62(6):2029–2036. doi: 10.1128/aem.62.6.2029-2036.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lemanceau P., Corberand T., Gardan L., Latour X., Laguerre G., Boeufgras J., Alabouvette C. Effect of Two Plant Species, Flax (Linum usitatissinum L.) and Tomato (Lycopersicon esculentum Mill.), on the Diversity of Soilborne Populations of Fluorescent Pseudomonads. Appl Environ Microbiol. 1995 Mar;61(3):1004–1012. doi: 10.1128/aem.61.3.1004-1012.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Martínez-Romero E., Segovia L., Mercante F. M., Franco A. A., Graham P., Pardo M. A. Rhizobium tropici, a novel species nodulating Phaseolus vulgaris L. beans and Leucaena sp. trees. Int J Syst Bacteriol. 1991 Jul;41(3):417–426. doi: 10.1099/00207713-41-3-417. [DOI] [PubMed] [Google Scholar]
  16. Nour S. M., Cleyet-Marel J. C., Normand P., Fernandez M. P. Genomic heterogeneity of strains nodulating chickpeas (Cicer arietinum L.) and description of Rhizobium mediterraneum sp. nov. Int J Syst Bacteriol. 1995 Oct;45(4):640–648. doi: 10.1099/00207713-45-4-640. [DOI] [PubMed] [Google Scholar]
  17. Nour S. M., Fernandez M. P., Normand P., Cleyet-Marel J. C. Rhizobium ciceri sp. nov., consisting of strains that nodulate chickpeas (Cicer arietinum L.). Int J Syst Bacteriol. 1994 Jul;44(3):511–522. doi: 10.1099/00207713-44-3-511. [DOI] [PubMed] [Google Scholar]
  18. Rome S., Fernandez M. P., Brunel B., Normand P., Cleyet-Marel J. C. Sinorhizobium medicae sp. nov., isolated from annual Medicago spp. Int J Syst Bacteriol. 1996 Oct;46(4):972–980. doi: 10.1099/00207713-46-4-972. [DOI] [PubMed] [Google Scholar]
  19. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  20. Segovia L., Young J. P., Martínez-Romero E. Reclassification of American Rhizobium leguminosarum biovar phaseoli type I strains as Rhizobium etli sp. nov. Int J Syst Bacteriol. 1993 Apr;43(2):374–377. doi: 10.1099/00207713-43-2-374. [DOI] [PubMed] [Google Scholar]
  21. Sullivan J. T., Eardly B. D., van Berkum P., Ronson C. W. Four unnamed species of nonsymbiotic rhizobia isolated from the rhizosphere of Lotus corniculatus. Appl Environ Microbiol. 1996 Aug;62(8):2818–2825. doi: 10.1128/aem.62.8.2818-2825.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sullivan J. T., Patrick H. N., Lowther W. L., Scott D. B., Ronson C. W. Nodulating strains of Rhizobium loti arise through chromosomal symbiotic gene transfer in the environment. Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8985–8989. doi: 10.1073/pnas.92.19.8985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Versalovic J., Koeuth T., Lupski J. R. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 1991 Dec 25;19(24):6823–6831. doi: 10.1093/nar/19.24.6823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991 Jan;173(2):697–703. doi: 10.1128/jb.173.2.697-703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Willems A., Collins M. D. Phylogenetic analysis of rhizobia and agrobacteria based on 16S rRNA gene sequences. Int J Syst Bacteriol. 1993 Apr;43(2):305–313. doi: 10.1099/00207713-43-2-305. [DOI] [PubMed] [Google Scholar]
  26. Yanagi M., Yamasato K. Phylogenetic analysis of the family Rhizobiaceae and related bacteria by sequencing of 16S rRNA gene using PCR and DNA sequencer. FEMS Microbiol Lett. 1993 Feb 15;107(1):115–120. doi: 10.1111/j.1574-6968.1993.tb06014.x. [DOI] [PubMed] [Google Scholar]
  27. de Bruijn F. J. Use of repetitive (repetitive extragenic palindromic and enterobacterial repetitive intergeneric consensus) sequences and the polymerase chain reaction to fingerprint the genomes of Rhizobium meliloti isolates and other soil bacteria. Appl Environ Microbiol. 1992 Jul;58(7):2180–2187. doi: 10.1128/aem.58.7.2180-2187.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. van Berkum P., Beyene D., Eardly B. D. Phylogenetic relationships among Rhizobium species nodulating the common bean (Phaseolus vulgaris L.). Int J Syst Bacteriol. 1996 Jan;46(1):240–244. doi: 10.1099/00207713-46-1-240. [DOI] [PubMed] [Google Scholar]
  29. van Berkum P. Evidence for a Third Uptake Hydrogenase Phenotype among the Soybean Bradyrhizobia. Appl Environ Microbiol. 1990 Dec;56(12):3835–3841. doi: 10.1128/aem.56.12.3835-3841.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. van Berkum P., Navarro R. B., Vargas A. A. Classification of the uptake hydrogenase-positive (Hup+) bean rhizobia as Rhizobium tropici. Appl Environ Microbiol. 1994 Feb;60(2):554–561. doi: 10.1128/aem.60.2.554-561.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES