Skip to main content

Identification of nodX, a gene that allows Rhizobium leguminosarum biovar viciae strain TOM to nodulate Afghanistan peas

Molecular and General Genetics MGG volume 212, pages 531–535 (1988)Cite this article

Summary

Gene(s) conferring the ability of Rhizobium leguminosarum biovar viciae strain TOM to nodulate primitive peas (cultivar Afghanistan) had been located in a 2.0 kb region of its sym plasmid, pRL5JI. In this DNA, a single open reading frame of 1101 bp, corresponding to a gene, nodX was found. nodX is downstream of nodJ which is present in strain TOM and also in the sym plasmid of a typical strain of this biovar. nodX specifies a hydrophobic protein (of Mr 41 036) with no clear similarity to other proteins in data bases. Mutations in nodX abolished nodulation of Afghanistan peas but not nodulation of commercial peas. nodX-lacZ fusions were used to show that transcription of nodX was activated by root exudates from both commercial and Afghanistan peas and by defined flavonoids. Exudate from Afghanistan peas activated nod genes of typical strains of R. leguminosarum biovar viciae which fail to nodulate these peas; thus, their failure to nodulate these primitive peas is not due to a lack of activation of their nod genes by exudate from Afghanistan peas. A homologue of nodX exists in R. leguminosarum biovar trifolii (which nodulates clover) but not in typical strains of R. leguminosarum biovar viciae.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  • Bankier AT, Barrell BG (1983) Shotgun DNA sequencing. In: Flavell RA (ed) Techniques in the Life Sciences B5 Nucleic Acid Biochemistry B508. Elsevier Scientific Publishers, Ireland, pp 1–34

    Google Scholar 

  • Beringer JE (1974) R factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198

    Google Scholar 

  • Beringer JE, Beynon JL, Buchanan-Wollaston AV, Johnston AWB (1978) Transfer of the drug-resis tance transposon Tn5 to Rhizobium. Nature (London) 276:633–634

    Google Scholar 

  • Beynon JL, Beringer JE, Johnston AWB (1980) Plasmids and host range in Rhizobium leguminosarum and Rhizobium phaseoli. J Gen Microbiol 120:421–429

    Google Scholar 

  • Brewin NJ, Beringer JE, Johnston AWB (1980) Plasmid mediated transfer of host-range specificity between two strains of Rhizobium leuminosarum. J Gen Microbiol 128:1817–1827

    Google Scholar 

  • Djordjevic MA, Schofield PR, Rolfe BG (1985) Tn5 mutagenesis of Rhizobium trifolii host-specific nodulation genes result in mutants with altered host-range ability. Mol Gen Genet 200:463–471

    Google Scholar 

  • Downie JA, Ma Q-S, Knight CD, Hombrecher G, Johnston AWB (1983) Cloning of the symbiotic region of Rhizobium leguminosarum: the nodulation genes are between the nitrogenase genes and a nifA-like gene. EMBO J 2:947–952

    Google Scholar 

  • Evans IJ, Downie JA (1986) The nodI gene product of Rhizobium leguminosarum is closely related to ATP-binding bacterial transport proteins; nucleotide sequence analysis of the nodI and nodJ genes. Gene 43:95–101

    Google Scholar 

  • Figurski D, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652

    Google Scholar 

  • Firmin JL, Wilson KE, Rossen L, Johnston AWB (1986) Flavonoid activation of nodulation genes of Rhizobium reversed by other compounds present in plants. Nature 324:90–92

    Google Scholar 

  • Friedman AN, Long SR, Brown SE, Buikema WJ, Ausubel FM (1982) Construction of a broad host-range cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene 13:227–237

    Google Scholar 

  • Götz R, Evans IJ, Downie JA, Johnston AWB (1985) Identification of the host-range DNA which allows Rhizobium leguminosarum strain TOM to nodulate cv. Afghanistan peas. Mol Gen Genet 201:296–300

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

  • Hombrecher G, Götz R, Dibb NJ, Downie JA, Johnston AWB, Brewin NJ (1984) Cloning and mutagenesis of nodulation genes from Rhizobium leguminosarum TOM, a strain with extended host range. Mol Gen Genet 194:293–298

    Google Scholar 

  • Hong GF (1981) A method for sequencing single-stranded cloned DNA in both directions. Biosci Rep 1:243–252

    Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward J, Schrempf H (1985) Genetic manipulation of Streptomyces. A laboratory manual. John Innes Foundation, Norwich

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Lamb JW, Hombrecher G, Johnston AWB (1982) Plasmid-determined nodulation and nitrogen-fixation abilities in Rhizobium phaseoli. Mol Gen Genet 186:449–452

    Google Scholar 

  • Lie TA (1978) Symbiotic specialisation in pea plants: the requirement of specific Rhizobium strains for peas from Afghanistan. Ann Appl Biol 88:462–465

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Rossen L, Shearman CA, Johnston AWB, Downie JA (1985) The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant exudate induces the nodABC genes. EMBO J 4:3369–3373

    Google Scholar 

  • Rossen L, Davis EO, Johnston AWB (1987) Plant induced expression of Rhizobium genes involved in host specificity and early stages of nodulation. Trends Biol Sci 12:430–435

    Google Scholar 

  • Ruvkun GB, Ausubel FM (1981) A general method for site-directed mutagenesis in prokaryotes. Nature (London) 289:85–88

    Google Scholar 

  • Sanger F, Nicklen S, Coulsen AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sanger F, Coulsen AR, Barrell BG, Smith AJH, Roe BA (1980) Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178

    Google Scholar 

  • Shearman CA, Rossen L, Johnston AWB, Downie JA (1986) The Rhizobium leguminosarum nodulation gene nodF encodes a polypeptide similar to acyl-carrier protein and is regulated by nodD plus a factor in pea root exudate. EMBO J 5:647–652

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrphoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Stachel SE, An G, Flores C, Nester EW (1985) A Tn3lacZ transposon for the random generation of β-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J 4:891–898

    Google Scholar 

  • Staden R (1984) Measurements of the effects that coding for a protein has on a DNA sequence and their use for finding genes. Nucleic Acids Res 12:551–567

    Google Scholar 

  • Winarno R, Lie TA (1979) Competition between Rhizobium strains in nodule formation: interactions between nodulating and non-nodulating strains. Plant Soil 51:135–142

    Google Scholar 

  • Young JPW, Johnston AWB, Brewin NJ (1982) A search for peas showing strain specificity for symbiotic Rhizobium leguminosarum. Heredity 48:197–201

    Google Scholar 

Download references

Author information

Author notes

  1. Ian J. Evans

    Present address: Celltech Limited, 244-250 Bath Road, SL1 4DY, Slough, UK

Affiliations

  1. John Innes Institute and A.F.R.C. Institute of Plant Science Research, Colney Lane, NR4 7UH, Norwich, UK

    Elaine O. Davis, Ian J. Evans & Andrew W. B. Johnston

Authors
  1. Elaine O. Davis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ian J. Evans
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrew W. B. Johnston
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by H. Hennecke

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Davis, E.O., Evans, I.J. & Johnston, A.W.B. Identification of nodX, a gene that allows Rhizobium leguminosarum biovar viciae strain TOM to nodulate Afghanistan peas. Mol Gen Genet 212, 531–535 (1988). https://doi.org/10.1007/BF00330860

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330860

Key words

  • Gene regulation
  • Nitrogen fixation
  • nod genes
  • Peas
  • Rhizobium