ABSTRACT
Symbiotic association of several genera of bacteria collectively called as rhizobia and plants belonging to the family Leguminosae (=Fabaceae) results in the process of biological nitrogen fixation, playing a key role in global N cycling, and also bringing relevant contributions to the agriculture. Bradyrhizobium is considered as the ancestral of all nitrogen-fixing rhizobial species, probably originated in the tropics. The genus encompasses a variety of diverse bacteria, but the diversity captured in the analysis of the 16S rRNA is often low. In this study, we analyzed twelve Bradyrhizobium strains selected from previous studies performed by our group for showing high genetic diversity in relation to the described species. In addition to the 16S rRNA, five housekeeping genes (recA, atpD, glnII, gyrB and rpoB) were analyzed in the MLSA (multilocus sequence analysis) approach. Analysis of each gene and of the concatenated housekeeping genes captured a considerably higher level of genetic diversity, with indication of putative new species. The results highlight the high genetic variability associated with Bradyrhizobium microsymbionts of a variety of legumes. In addition, the MLSA approach has proved to represent a rapid and reliable method to be employed in phylogenetic and taxonomic studies, speeding the identification of the still poorly known diversity of nitrogen-fixing rhizobia in the tropics.
Subject(s)
Base Sequence , Bradyrhizobium/genetics , Nitrogen Fixation/genetics , Genetic Variation , In Vitro Techniques , Phylogeny , Polymerase Chain Reaction , RNA , Reverse Transcriptase Polymerase Chain Reaction/methods , Rhizobium leguminosarum/genetics , Methods , Symbiosis/genetics , Tropical EcosystemABSTRACT
High temperatures can affect the survival, establishment and symbiotic properties of "Rhizobium" strains. Bean nodulating "Rhizobium" strains are considered particularly sensitive because on this strains genetic recombinations and/or deletions occur frequently, thus compromising the use of these bacteria as inoculants. In this study "R. tropici" and "R. leguminosarum" bv. "phaseoli" strains isolated from Cerrado soils were exposed to thermal stress and the strains' growth, survival and symbiotic relationships as well as alterations in their genotypic and phenotypic were analysed. After successive thermal shocks at 45ºC for four hours, survival capacity appeared to be strain-specifc, independent of thermo-tolerance and was more apparent in "R. tropici" strains (with the exception of FJ2.21) were more stable than "R. leguminosarum" bv. "phaseoli" strains because no significant phenotypic alterations were observed following thermal treatments and they maintained their original genotypic pattern after innoculation in plants.