RESUMO
DNA-DNA hybridizations were performed between Bradyrhizobium strains, isolated mainly from Faidherbia albida and Aeschynomene species, as well as Bradyrhizobium reference strains. Results indicated that the genus Bradyrhizobium consists of at least 11 genospecies, I to XI. The genospecies formed four subgeneric groups that were more closely related to each other (>40% DNA hybridization) than to other genospecies (<40% DNA hybridization): (i) genospecies I (Bradyrhizobium japonicum), III (Bradyrhizobium liaoningense), IV and V; (ii) genospecies VI and VIII; (iii) genospecies VII and IX; and (iv) genospecies II (Bradyrhizobium elkanii), X and XI. Photosynthetic Aeschynomene isolates were found to belong to at least two distinct genospecies in one subgeneric group. DNA-DNA hybridization data are compared with data from amplified fragment length polymorphism analysis and 165-23S rDNA spacer sequence analysis.
Assuntos
Bradyrhizobium/genética , DNA Bacteriano/genética , Bradyrhizobium/classificação , Bradyrhizobium/isolamento & purificação , Bradyrhizobium/metabolismo , Fabaceae/microbiologia , Hibridização de Ácido Nucleico , Fotossíntese , Polimorfismo Genético , Especificidade da EspécieRESUMO
We examined the genotypic diversity of 64 Bradyrhizobium strains isolated from nodules from 27 native leguminous plant species in Senegal (West Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia, which play an ecological role and have agronomic potential in arid regions. The strains were characterized by intergenic spacer (between 16S and 23S rRNA genes) PCR and restriction fragment length polymorphism (IGS PCR-RFLP) and amplified fragment length polymorphism (AFLP) fingerprinting analyses. Fifty-three reference strains of the different Bradyrhizobium species and described groups were included for comparison. The strains were diverse and formed 27 groups by AFLP and 16 groups by IGS PCR-RFLP. The sizes of the IGS PCR products from the Bradyrhizobium strains that were studied varied from 780 to 1,038 bp and were correlated with the IGS PCR-RFLP results. The grouping of strains was consistent by the three methods AFLP, IGS PCR-RFLP, and previously reported 16S amplified ribosomal DNA restriction analysis. For investigating the whole genome, AFLP was the most discriminative technique, thus being of particular interest for future taxonomic studies in Bradyrhizobium, for which DNA is difficult to obtain in quantity and quality to perform extensive DNA:DNA hybridizations.
Assuntos
Bradyrhizobium/classificação , Impressões Digitais de DNA/métodos , DNA Intergênico , Fabaceae/microbiologia , Plantas Medicinais , Técnicas de Tipagem Bacteriana , Bradyrhizobium/genética , Análise por Conglomerados , DNA Bacteriano/genética , DNA Ribossômico/genética , Genótipo , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , SenegalRESUMO
The diversity of Bradyrhizobium isolates from Faidherbia albida and Aeschynomenee species was assessed using AFLP analysis, a high-resolution genomic fingerprinting technique. Reference strains from Bradyrhizobium japonicum, Bradyrhizobium elkanii and Bradyrhizobium liaoningense were included for comparison. At a similarity level of 50%, a total of 34 different groups were obtained by cluster analysis of the genomic fingerprints. Four of these clusters correspond to the three reference species, demonstrating the large diversity of the isolates studied. Comparison with other data demonstrates that AFLP has a higher resolution than restriction analysis of 16S rRNA genes, SDS-PAGE analysis of proteins and phenotypic analysis. Results of the latter two methods showed little correspondence with the genotypic data.
Assuntos
Acacia/microbiologia , Bradyrhizobium/classificação , Impressões Digitais de DNA/métodos , Fabaceae/microbiologia , Variação Genética , Genoma Bacteriano , Plantas Medicinais , Bradyrhizobium/genética , Bradyrhizobium/isolamento & purificação , Genótipo , Polimorfismo de Fragmento de RestriçãoRESUMO
We isolated 71 slow-growing bacterial strains from nodules of 27 native leguminous plants species in Senegal (West-Africa) belonging to the genera Abrus, Alysicarpus, Bryaspis, Chamaecrista, Cassia, Crotalaria, Desmodium, Eriosema, Indigofera, Moghania, Rhynchosia, Sesbania, Tephrosia, and Zornia playing an ecological role and having agronomic potential in arid regions. The isolates were characterised by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified 16S rDNA and comparative SDS-PAGE of whole-cell proteins; reference strains of the different known rhizobial species and groups were included as references. We conclude that these nodule isolates are diverse, and form several phylogenetic subgroups inside Bradyrhizobium. Nodulation tests performed on 5 plant species demonstrated host specificity among the strains studied.
