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Microbial Ecology

, Volume 46, Issue 1, pp 113–121 | Cite as

Genetic Characterization of the Nitrate Reducing Community Based on narG Nucleotide Sequence Analysis

  • D. Chèneby
  • S. Hallet
  • M. Mondon
  • F. Martin-Laurent
  • J. C. Germon
  • L. PhilippotEmail author
Article

Abstract

The ability of facultative anerobes to respire nitrate has been ascribed mainly to the activity of a membrane-bound nitrate reductase encoded by the narGHJI operon. Respiratory nitrate reduction is the first step of the denitrification pathway, which is considered as an important soil process since it contributes to the global cycling of nitrogen. In this study, we employed direct PCR, cloning, and sequencing of narG gene fragments to determine the diversity of nitrate-reducing bacteria occurring in soil and in the maize rhizosphere. Libraries containing 727 clones in total were screened by restriction fragment analysis. Phylogenetic analysis of 128 narG sequences separated the clone families into two main groups that represent the Gram-positive and Gram-negative nitrate-reducing bacteria. Novel narG lineages that branch distinctly from all currently known membrane bound nitrate-reductase encoding genes were detected within the Gram-negative branch. All together, our results revealed a more complex nitrate-reducing community than did previous culture-based studies. A significant and consistent shift in the relative abundance of the nitrate-reducing groups within this functional community was detected in the maize rhizosphere. Thus a substantially higher abundance of the dominant clone family and a lower diversity index were observed in the rhizosphere compared to the unplanted soil, suggesting that a bacterial group has been specifically selected within the nitrate-reducing community. Furthermore, restriction fragment length polymorphism analysis of cloned narG gene fragments proved to be a powerful tool in evaluating the structure and the diversity of the nitrate-reducing community and community shifts therein.

Keywords

Nitrate Reductase Rhizospheric Soil Functional Community Unplanted Soil Restriction Fragment Length Polymorphism Type 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported by the PNSE program “rhizosphere bioreactor” of the Institut National des Sciences de l’Univers and the Burgundy region program B03725.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • D. Chèneby
    • 1
  • S. Hallet
    • 1
  • M. Mondon
    • 1
  • F. Martin-Laurent
    • 1
  • J. C. Germon
    • 1
  • L. Philippot
    • 1
    Email author
  1. 1.UMR A111 Microbiologie des Sols, GéosolsInstitut National de la Recherche Agronomique, 17, rue Sully, B.P. 86510, 21065 Dijon CedexFrance

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