Rhizobium lusitanum sp nov a bacterium that nodulates Phaseolus vulgaris

The species Phaseolus vulgaris is a promiscuous legume nodulated by several species of the family Rhizobiaceae. During a study of rhizobia nodulating this legume in Portugal, we isolated several strains that nodulate P. vulgaris effectively and also Macroptilium atropurpureum and Leucaena leucocephala, but they form ineffective nodules in Medicago sativa. According to phylogenetic analysis of the 16S rRNA gene sequence, the strains from this study belong to the genus Rhizobium, with Rhizobium rhizogenes and Rhizobium tropici as the closest related species, with 99.9 and 99.2% similarity, respectively, between the type strains of these species and strain P1-7(T). The nodD and nifH genes carried by strain P1-7(T) are phylogeneticafly related to those of other species nodulating Phaseolus. This strain does not carry virulence genes present in the type strain of R. rhizogenes, ATCC 11325(T). Analysis of the recA and atpD genes confirms this phylogenetic arrangement, showing low similarity with respect to those of R. rhizogenes ATCC 11325(T) (91.9 and 94.1% similarity, respectively) and R. tropici IIB CAT 899(T) (90.6 % and 91.8 % similarity, respectively). The intergenic spacer (ITS) of the strains from this study is phylogenetically divergent from those of R. rhizogenes ATCC 11235(T) and R. tropici CAT 899(T), with 85.9 and 82-8 % similarity, respectively, with respect to strain P1-7(T). The tRNA profile and two-primer random amplified polymorphic DNA pattern of strain P1-7(T) are also different from those of R. rhizogenes ATCC 11235(T) and R. tropici CAT 899(T). The strains isolated in this study can be also differentiated from R. rhizogenes and R. tropici by several phenotypic characteristics. The results of DNA-DNA hybridization showed means of 28 and 25 % similarity between strain P1-7(T) and R. rhizogenes ATCC 11235(T) and R. tropici CIAT 899(T), respectively. All these data showed that the strains isolated in this study belong to a novel species of the genus Rhizobium, for which we propose the name Rhizobium lusitanum sp. nov.; the type strain is P1-7(T) (= LMG 22705(T) = CECT 7016(T)).