Diversity and phylogeny of rhizobial strains isolated from Lotus uliginosus grown in Uruguayan soils

Lotus uliginosus is generally nodulated by rhizobia of the genus Bradyrhizobium when used for improvement of Uruguayan pastures. The genetic diversity and phylogenetic relationships of 111 isolates from nodules of L. uliginosus collected from four fields with or without prior inoculation history were analyzed in this study. Genetic diversity estimated by ERIC-PCR revealed 75 different genomic fingerprints, and showed a relatively greater value compared with other methods and varied by soil type. 16S ribosomal RNA gene RFLP analysis revealed three different ribogroups, A, B and C, with 71 isolates in ribogroup A, three isolates in ribogroup B and only one in ribogroup C. Phylogenetic analyses based on 16S RNA gene sequences, ITS, as well as atpD, recA and glnII gene sequences indicated that ribogroup A strains were affiliated with B. japonicum bv. genistearum strains. The three isolates in ribogroup B did not clearly associate with any Bradyrhizobium species described previously and could represent a novel species within this genus. Unlike B. japonicum strains these isolates were able to nodulate and fix nitrogen with other Lotus species as well as with Spartium, a leguminous shrub. The unique isolate in ribogroup C clustered with Mesorhizobium and appeared genetically and phenotypically related to broad host-range Mesorhizobium sp. NZP2037. Our data suggest that Uruguayan soils contain native or naturalized bradyrhizobia that are able to nodulate L. uliginosus as efficiently as the commercial strain NZP2309 but could have adaptive advantages making them more suitable for inoculant purposes.