Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1985 Jul;50(1):41–44. doi: 10.1128/aem.50.1.41-44.1985

Evidence for plasmid- and chromosome-borne multiple nif genes in Rhizobium fredii.

W M Barbour, J N Mathis, G H Elkan
PMCID: PMC238570  PMID: 2992376

Abstract

Rhizobium fredii is a fast-growing rhizobium isolated from the primitive Chinese soybean cultivar Peking and from the wild soybean Glycine soja. This rhizobium harbors nif genes on 150- to 200-megadalton plasmids. By passage on acridine orange plates, we obtained a mutant of R. fredii USDA 206 cured of the 197-megadalton plasmid (USDA 206C) which carries both nif and nod genes. This strain, however, has retained its symbiotic effectiveness. Probing EcoRI digests of wild-type and cured plasmid DNA with a 2.2-kilobase nif DH fragment from Rhizobium meliloti has shown four homologous fragments in the wild-type strain (4.2, 4.9, 10, and 11 kilobases) and two fragments in the cured strain (4.2 and 10 kilobases). EcoRI digests of total DNA show four major bands of homology (4.2, 4.9, 5.8, and 13 kilobases) in both the wild-type and cured strains. The presence of major bands of homology in the total DNA not present in the plasmid DNA indicated chromosomal nif genes. Probing of HindIII digests of total and plasmid DNA led to the same conclusion. Hybridization to the smaller plasmids of USDA 206 and USDA 206C showed the presence of nif genes on at least one of these plasmids, explaining the nif homology in the USDA 206C plasmid digests.

Full text

PDF
41

Images in this article

42Image
on p.42
42Image
on p.42
43Image
on p.43

Selected References

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

  1. 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]
  2. Broughton W. J., Heycke N., Z A H. M., Pankhurst C. E. Plasmid-linked nif and "nod" genes in fast-growing rhizobia that nodulate Glycine max, Psophocarpus tetragonolobus, and Vigna unguiculata. Proc Natl Acad Sci U S A. 1984 May;81(10):3093–3097. doi: 10.1073/pnas.81.10.3093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  4. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  5. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Keyser H. H., Bohlool B. B., Hu T. S., Weber D. F. Fast-growing rhizobia isolated from root nodules of soybean. Science. 1982 Mar 26;215(4540):1631–1632. doi: 10.1126/science.215.4540.1631. [DOI] [PubMed] [Google Scholar]
  7. Krol A. J., Hontelez J. G., Van den Bos R. C., Van Kammen A. Expression of large plasmids in the endosymbiotic form of Rhizobium leguminosarum. Nucleic Acids Res. 1980 Oct 10;8(19):4337–4347. doi: 10.1093/nar/8.19.4337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Leonard L. T. A Simple Assembly for Use in the Testing of Cultures of Rhizobia. J Bacteriol. 1943 Jun;45(6):523–527. doi: 10.1128/jb.45.6.523-527.1943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Masterson R. V., Russell P. R., Atherly A. G. Nitrogen fixation (nif) genes and large plasmids of Rhizobium japonicum. J Bacteriol. 1982 Nov;152(2):928–931. doi: 10.1128/jb.152.2.928-931.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mathis J. N., Barbour W. M., Elkan G. H. Effect of Sym Plasmid Curing on Symbiotic Effectiveness in Rhizobium fredii. Appl Environ Microbiol. 1985 Jun;49(6):1385–1388. doi: 10.1128/aem.49.6.1385-1388.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Norgard M. V. Rapid and simple removal of contaminating RNA from plasmid DNA without the use of RNase. Anal Biochem. 1981 May 1;113(1):34–42. doi: 10.1016/0003-2697(81)90040-3. [DOI] [PubMed] [Google Scholar]
  12. Prakash R. K., Atherly A. G. Reiteration of genes involved in symbiotic nitrogen fixation by fast-growing Rhizobium japonicum. J Bacteriol. 1984 Nov;160(2):785–787. doi: 10.1128/jb.160.2.785-787.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Prakash R. K., Schilperoort R. A., Nuti M. P. Large plasmids of fast-growing rhizobia: homology studies and location of structural nitrogen fixation (nif) genes. J Bacteriol. 1981 Mar;145(3):1129–1136. doi: 10.1128/jb.145.3.1129-1136.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sadowsky M. J., Bohlool B. B. Possible involvement of a megaplasmid in nodulation of soybeans by fast-growing rhizobia from china. Appl Environ Microbiol. 1983 Oct;46(4):906–911. doi: 10.1128/aem.46.4.906-911.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sloger C. Symbiotic effectiveness and n(2) fixation in nodulated soybean. Plant Physiol. 1969 Dec;44(12):1666–1668. doi: 10.1104/pp.44.12.1666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  17. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]
  19. Zurkowski W., Hoffman M., Lorkiewicz Z. Effect of acriflavine and sodium dodecyl sulphate on infectiveness of Rhizobium trifolii. Acta Microbiol Pol A. 1973;5(2):55–60. [PubMed] [Google Scholar]

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

RESOURCES