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Molecular cloning of thentrA gene of the broad host-rangeRhizobium sp. NGR234, and phenotypes of a site-directed mutant

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Summary

The clonedntrA (rpoN) gene andntrA mutants ofRhizobium meliloti were used to isolate the homologous gene from the broad-host rangeRhizobium sp. NGR234 by hybridization and interspecies complementation. The NGR234 locus was analyzed by deletion and insertional mutagenesis. A site-directedntrA mutant, NGR234rn1, was made with an interposon, GmI, and its phenotype was examined ex planta and in symbiosis. NGR234rn1 formed Fix− nodules on six genera tested from among its legume hosts, including both indeterminate and determinate nodule-type plants. Formation of nodules onMacroptilium was delayed, and expression of anR. meliloti nodABC-lacZ fusion was reduced by the mutant allele.

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Abbreviations

NTRA, NTRC, NIFA:

protein products ofntrA,ntrC,nifA genes

Tc:

tetracycline

Nm:

neomycin

Rif:

ritampicin

Gm:

gentamicin

Sp:

spectinomycin

Km:

kanamycin

Cm:

chloramphenicol

Ap:

ampicillin

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Authors and Affiliations

  1. Laboratoire de Biologie Moléculaire des Plantes Supérieures, University of Geneva, 1 chemin de l’Impératrice, CH-1292, Chambésy-Genève, Switzerland

    John Stanley, Jan van Slooten, David N. Dowling & William J. Broughton

  2. Department of Biology, McMaster University, L8S 4K1, Hamilton, Ontario, Canada

    Turlough Finan

Authors
  1. John Stanley
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  2. Jan van Slooten
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  3. David N. Dowling
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  4. Turlough Finan
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  5. William J. Broughton
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Communicated by H. Hennecke

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Stanley, J., van Slooten, J., Dowling, D.N. et al. Molecular cloning of thentrA gene of the broad host-rangeRhizobium sp. NGR234, and phenotypes of a site-directed mutant. Molec. Gen. Genet. 217, 528–532 (1989). https://doi.org/10.1007/BF02464927

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  • DOI: https://doi.org/10.1007/BF02464927

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