Rhizobium gone native: Unexpected plasmid stability of indigenous Rhizobium leguminosarum

  1. Jennifer J. Wernegreen*,,
  2. Ethelynda E. Harding, and
  3. Margaret A. Riley*
  1. *Department of Biology, Yale University, New Haven, CT 06511; and Department of Biology, California State University, Fresno, CA 93740-0073

Abstract

Lateral transfer of bacterial plasmids is thought to play an important role in microbial evolution and population dynamics. However, this assumption is based primarily on investigations of medically or agriculturally important bacterial species. To explore the role of lateral transfer in the evolution of bacterial systems not under intensive, human-mediated selection, we examined the association of genotypes at plasmid-encoded and chromosomal loci of native Rhizobium, the nitrogen-fixing symbiont of legumes. To this end, Rhizobium leguminosarum strains nodulating sympatric species of native Trifolium were characterized genetically at plasmid-encoded symbiotic (sym) regions (nodulation AB and nodulation CIJT loci) and a repeated chromosomal locus not involved in the symbiosis with legumes. Restriction fragment length polymorphism analysis was used to distinguish genetic groups at plasmid and chromosomal loci. The correlation between major sym and chromosomal genotypes and the distribution of genotypes across host plant species and sampling location were determined using χ2 analysis. In contrast to findings of previous studies, a strict association existed between major sym plasmid and chromosomal genetic groups, suggesting a lack of successful sym plasmid transfer between major Rhizobium chromosomal types. These data indicate that previous observations of sym plasmid transfer in agricultural settings may seriously overestimate the rates of successful conjugation in systems not impacted by human activities. In addition, a nonrandom distribution of Rhizobium genotypes across host plant species and sampling site demonstrates the importance of both factors in shaping Rhizobium population dynamics.

Footnotes

  • To whom reprint requests should be addressed. e-mail: werjen{at}minerva.cis.yale.edu.

  • Allan Campbell, Stanford University, Stanford, CA

  • Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. U72626U72626 and U72627U72627).

  • ABBREVIATIONS:
    sym plasmid,
    symbiotic plasmid;
    nodAB,
    nodulation AB loci;
    nodCIJT,
    nodulation CIJT loci;
    16S rRNA,
    16S ribosomal RNA gene;
    RFLP,
    restriction fragment length polymorphism
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