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Journal of Bacteriology, December 2003, p. 7266-7272, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7266-7272.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Legume Symbiotic Nitrogen Fixation by ß-Proteobacteria Is Widespread in Nature

Wen-Ming Chen,1 Lionel Moulin,2,{dagger} Cyril Bontemps,2,3 Peter Vandamme,4 Gilles Béna,2 and Catherine Boivin-Masson2,3*

Laboratory of Microbiology, Department of Seafood Science, National Kaohsiung Institute of Marine Technology, Kaohsiung City 811, Taiwan,1 LSTM, IRD-INRA-CIRAD-ENSAM, TA 10/J, Baillarguet, 34 398 Montpellier Cedex 5,2 Laboratoire des Interactions Plantes Micro-Organismes, INRA-CNRS, 31 326 Castanet-Tolosan Cedex, France,3 Laboratorium voor Microbiologie, Universiteit Gent, B-9000 Ghent, Belgium4

Received 20 June 2003/ Accepted 24 September 2003

Following the initial discovery of two legume-nodulating Burkholderia strains (L. Moulin, A. Munive, B. Dreyfus, and C. Boivin-Masson, Nature 411:948-950, 2001), we identified as nitrogen-fixing legume symbionts at least 50 different strains of Burkholderia caribensis and Ralstonia taiwanensis, all belonging to the ß-subclass of proteobacteria, thus extending the phylogenetic diversity of the rhizobia. R. taiwanensis was found to represent 93% of the Mimosa isolates in Taiwan, indicating that ß-proteobacteria can be the specific symbionts of a legume. The nod genes of rhizobial ß-proteobacteria (ß-rhizobia) are very similar to those of rhizobia from the {alpha}-subclass ({alpha}-rhizobia), strongly supporting the hypothesis of the unique origin of common nod genes. The ß-rhizobial nod genes are located on a 0.5-Mb plasmid, together with the nifH gene, in R. taiwanensis and Burkholderia phymatum. Phylogenetic analysis of available nodA gene sequences clustered ß-rhizobial sequences in two nodA lineages intertwined with {alpha}-rhizobial sequences. On the other hand, the ß-rhizobia were grouped with free-living nitrogen-fixing ß-proteobacteria on the basis of the nifH phylogenetic tree. These findings suggest that ß-rhizobia evolved from diazotrophs through multiple lateral nod gene transfers.

* Corresponding author. Mailing address: Laboratoire des Interactions Plantes-Microorganismes, INRA-CNRS, BP 27, 31 326 Castanet-Tolosan Cedex, France. Phone: (33) 5 61 28 54 49. Fax: (33) 5 61 28 50 61. E-mail: boivin{at}toulouse.inra.fr.

{dagger} Present address: Department of Biology 3, University of York, P.O. Box 373, York YO10 5YW, United Kingdom.

Journal of Bacteriology, December 2003, p. 7266-7272, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7266-7272.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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