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

Wen-Ming Chen,¹ Lionel Moulin,^2, Cyril Bontemps,^2,3 Peter Vandamme,⁴ Gilles Béna,² and Catherine Boivin-Masson^2,3*

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 -subclass (-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 -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.

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

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