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1Laboratoire de Recherche sur les Symbiotes des Racines, I.N.R.A-E.N.S.A.M., 34060 Montpellier cedex 01, France
2Laboratoire d'Ecologie Microbienne du sol UMR CNRS 5557, Université Lyon I, 69622 Villeurbanne cedex, France
3Unite de Formation et de Recherche de Science du Sol, 34060 Montpellier cedex 01, France
* Corresponding author. Mailing address: Laboratoire d'Ecologie Microbienne du Sol UMR CNRS 5557, Université Lyon I, Bàtiment 741, 43 Bd. du 11 Novembre 1918, 69622 Villeurbanne cedex, France. Fax: 72-43-12-23. Electronic mail address: sr{at}geometrie.univ-lyonl.fr.
ABSTRACT
The taxonomic position of isolates of a new genomic species (designated genomic species 2) obtained from several annual Medicago species and originating from different geographical locations was established through the results of phenotypic tests (including the of auxanographic and biochemical tests and symbiotic properties) and 16S rRNA phylogenetic inferences. A comparison of the complete 16S rRNA sequence of a representative of genomic species 2 (strain A 321T [T = type strain]) with the 16S rRNA sequences of other members of the Rhizobiaceae and closely related taxa showed that genomic species 2 was phylogenetically related to Sinorhizobium meliloti, Sinorhizobium fredii, Sinorhizobium saheli, and Sinorhizobium teranga. The levels of sequence similarity and observed numbers of nucleotide substitutions in Sinorhizobium strains indicated that A 321T and S. meliloti exhibited the highest level of sequence similarity (99.7%), with four nucleotide substitutions and one deletion. The results of a numerical analysis based on data from 63 auxanographic and biochemical tests clearly separated genomic species 2 isolates from S. meliloti. Genomic species 2 isolates nodulated and fixed nitrogen with Medicago polymorpha, whereas S. meliloti isolates were ineffective and formed rudimentary nodules on this host plant. On the basis of phenotypic and 16S sequence analysis data, genomic species 2 isolates cannot be assigned to a previously described species. We propose that these isolates belong to a new species, Sinorhizobium medicae.
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