Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Nov;171(11):6161–6168. doi: 10.1128/jb.171.11.6161-6168.1989

Genes involved in lipopolysaccharide production and symbiosis are clustered on the chromosome of Rhizobium leguminosarum biovar viciae VF39.

U B Priefer 1
PMCID: PMC210485  PMID: 2553672

Abstract

Four mutants of Rhizobium leguminosarum biovar viciae VF39 altered in lipopolysaccharide (LPS) synthesis were isolated upon random Tn5 mutagenesis. These mutants produced matt colonies on TY medium and showed autoagglutination and loss of motility. On sodium dodecyl sulfate-polyacrylamide gels, they lacked a slow-migrating carbohydrate band, corresponding to the complete LPS (LPSI). All four mutants formed small white nodules on Vicia hirsuta. These nodules were infected but showed no nitrogen-fixing activity and senesced prematurely. Three of the mutants were complemented by a wild-type cosmid to synthesis of normal LPS and induction of nitrogen-fixing nodules. By hybridization and in vivo cloning experiments, the mutations were mapped within different EcoRI fragments which could be localized on the VF39 chromosome. Cross-complementation analyses revealed that the three mutants were affected in different transcriptional units. The results indicate that a cluster of genes necessary for LPSI production and symbiotic efficiency is located within a defined region of 20 kilobases on the R. leguminosarum bv. viciae chromosome.

Full text

PDF
6161

Images in this article

6163Image
on p.6163
6164Image
on p.6164
6164Image
on p.6164
6165Image
on p.6165

Selected References

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

  1. Aguilar O. M., Kapp D., Pühler A. Characterization of a Rhizobium meliloti fixation gene (fixF) located near the common nodulation region. J Bacteriol. 1985 Oct;164(1):245–254. doi: 10.1128/jb.164.1.245-254.1985. [DOI] [PMC free article] [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. Carlson R. W. Heterogeneity of Rhizobium lipopolysaccharides. J Bacteriol. 1984 Jun;158(3):1012–1017. doi: 10.1128/jb.158.3.1012-1017.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlson R. W., Kalembasa S., Turowski D., Pachori P., Noel K. D. Characterization of the lipopolysaccharide from a Rhizobium phaseoli mutant that is defective in infection thread development. J Bacteriol. 1987 Nov;169(11):4923–4928. doi: 10.1128/jb.169.11.4923-4928.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carlson R. W., Shatters R., Duh J. L., Turnbull E., Hanley B., Rolfe B. G., Djordjevic M. A. The Isolation and Partial Characterization of the Lipopolysaccharides from Several Rhizobium trifolii Mutants Affected in Root Hair Infection. Plant Physiol. 1987 Jun;84(2):421–427. doi: 10.1104/pp.84.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carlson R. W., Yadav M. Isolation and partial characterization of the extracellular polysaccharides and lipopolysaccharides from fast-growing Rhizobium japonicum USDA 205 and its Nod- mutant, HC205, which lacks the symbiotic plasmid. Appl Environ Microbiol. 1985 Nov;50(5):1219–1224. doi: 10.1128/aem.50.5.1219-1224.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cava J. R., Elias P. M., Turowski D. A., Noel K. D. Rhizobium leguminosarum CFN42 genetic regions encoding lipopolysaccharide structures essential for complete nodule development on bean plants. J Bacteriol. 1989 Jan;171(1):8–15. doi: 10.1128/jb.171.1.8-15.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Grönger P., Manian S. S., Reiländer H., O'Connell M., Priefer U. B., Pühler A. Organization and partial sequence of a DNA region of the Rhizobium leguminosarum symbiotic plasmid pRL6JI containing the genes fixABC, nifA, nifB and a novel open reading frame. Nucleic Acids Res. 1987 Jan 12;15(1):31–49. doi: 10.1093/nar/15.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Halverson L. J., Stacey G. Signal exchange in plant-microbe interactions. Microbiol Rev. 1986 Jun;50(2):193–225. doi: 10.1128/mr.50.2.193-225.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hynes M. F., Simon R., Pühler A. The development of plasmid-free strains of Agrobacterium tumefaciens by using incompatibility with a Rhizobium meliloti plasmid to eliminate pAtC58. Plasmid. 1985 Mar;13(2):99–105. doi: 10.1016/0147-619x(85)90062-9. [DOI] [PubMed] [Google Scholar]
  13. Johnson K. G., Perry M. B. Improved techniques for the preparation of bacterial lipopolysaccharides. Can J Microbiol. 1976 Jan;22(1):29–34. doi: 10.1139/m76-004. [DOI] [PubMed] [Google Scholar]
  14. Klipp W., Masepohl B., Pühler A. Identification and mapping of nitrogen fixation genes of Rhodobacter capsulatus: duplication of a nifA-nifB region. J Bacteriol. 1988 Feb;170(2):693–699. doi: 10.1128/jb.170.2.693-699.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Maier R. C., Norris H. J. Glassy cell carcinoma of the cervix. Obstet Gynecol. 1982 Aug;60(2):219–224. [PubMed] [Google Scholar]
  17. Maier R. J., Brill W. J. Involvement of Rhizobium japonicum O antigen in soybean nodulation. J Bacteriol. 1978 Mar;133(3):1295–1299. doi: 10.1128/jb.133.3.1295-1299.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Noel K. D., Vandenbosch K. A., Kulpaca B. Mutations in Rhizobium phaseoli that lead to arrested development of infection threads. J Bacteriol. 1986 Dec;168(3):1392–1401. doi: 10.1128/jb.168.3.1392-1401.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Priefer U. B., Kalinowski J., Rüger B., Heumann W., Pühler A. ISR1, a transposable DNA sequence resident in Rhizobium class IV strains, shows structural characteristics of classical insertion elements. Plasmid. 1989 Mar;21(2):120–128. doi: 10.1016/0147-619x(89)90055-3. [DOI] [PubMed] [Google Scholar]
  20. Simon R. High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet. 1984;196(3):413–420. doi: 10.1007/BF00436188. [DOI] [PubMed] [Google Scholar]
  21. Simon R., O'Connell M., Labes M., Pühler A. Plasmid vectors for the genetic analysis and manipulation of rhizobia and other gram-negative bacteria. Methods Enzymol. 1986;118:640–659. doi: 10.1016/0076-6879(86)18106-7. [DOI] [PubMed] [Google Scholar]
  22. Simon R., Quandt J., Klipp W. New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in gram-negative bacteria. Gene. 1989 Aug 1;80(1):161–169. doi: 10.1016/0378-1119(89)90262-x. [DOI] [PubMed] [Google Scholar]
  23. Smit G., Kijne J. W., Lugtenberg B. J. Roles of flagella, lipopolysaccharide, and a Ca2+-dependent cell surface protein in attachment of Rhizobium leguminosarum biovar viciae to pea root hair tips. J Bacteriol. 1989 Jan;171(1):569–572. doi: 10.1128/jb.171.1.569-572.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  26. Wolpert J. S., Albersheim P. Host-symbiont interactions. I. The lectins of legumes interact with the o-antigen-containing lipopolysaccharides of their symbiont Rhizobia. Biochem Biophys Res Commun. 1976 Jun 7;70(3):729–737. doi: 10.1016/0006-291x(76)90653-7. [DOI] [PubMed] [Google Scholar]
  27. Wood W. B. Host specificity of DNA produced by Escherichia coli: bacterial mutations affecting the restriction and modification of DNA. J Mol Biol. 1966 Mar;16(1):118–133. doi: 10.1016/s0022-2836(66)80267-x. [DOI] [PubMed] [Google Scholar]
  28. de Maagd R. A., Rao A. S., Mulders I. H., Goosen-de Roo L., van Loosdrecht M. C., Wijffelman C. A., Lugtenberg B. J. Isolation and characterization of mutants of Rhizobium leguminosarum bv. viciae 248 with altered lipopolysaccharides: possible role of surface charge or hydrophobicity in bacterial release from the infection thread. J Bacteriol. 1989 Feb;171(2):1143–1150. doi: 10.1128/jb.171.2.1143-1150.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES