RESUMO
Heavy metals accumulation in the environment has led to a decrease in the capacity of ecosystems to sustain life as human, animal and plant health is threatened. To remedy this problem, rhizoremediation has been suggested as a solution. Legumes and rhizobia symbiotic association has captivated attention due to its involvement in the restoration of heavy-metal-contaminated sites. Thus, the aim of this study was to isolate and characterize the strains nodulating Calicotome spinosa plant that naturally occurred in two Algerian mercury mines. Fifty-four bacterial strains were isolated, then grouped into sixteen distinct BOX-PCR patterns and were genetically identified as belonging to the Bradyrhizobium genus. The studied strains were able to induce nodules on Retama monosperma, R. reatam, Lupinus albus, while no nodulation was observed in Glycine max, their symbiotic capacity was confirmed by amplifying the nodC gene. The phylogenetic analysis based on the nodC has grouped this Bradyrhizobium strains to either symbiovar genistearum or retamae. The isolates revealed diversity in terms of NaCl; pH tolerance, and phosphate solubilization. Production of siderophores was negative for these strains. All the isolated Bradyrhizobium were tolerant to both Zn and Pb in contrast they were sensitive to Cu and Cd. Interestingly, 43% of strains were tolerant to high Hg levels. Hence, some strains displayed multiple tolerances to heavy metals. Therefore, this is the first time we identify Bradyrhizobium strains originating from a North African mercury mine. This study could help to select mercury and other heavy metal-tolerant rhizobia showing an interesting potential to be used as inoculants to remediate the heavy metal soil accumulation.
Assuntos
Bradyrhizobium , Mercúrio , Metais Pesados , Argélia , Bradyrhizobium/genética , DNA Bacteriano , Ecossistema , Humanos , Mercúrio/toxicidade , Metais Pesados/toxicidade , Mineração , Filogenia , RNA Ribossômico 16S , Nódulos Radiculares de Plantas , Análise de Sequência de DNA , SimbioseRESUMO
Fifty-two slow-growing strains were isolated from root nodules of Calicotome spinosa grown in the Northeast of Algeria and grouped in 24 rep-PCR clusters. One representative strain for each profile was further phylogenetically characterized. The nearly complete 16S rRNA gene sequence indicated that all strains were affiliated to Bradyrhizobium. Multi-Locus Sequence Analysis (MLSA) of the atpD, glnII and recA genes and of the 16S-23S rRNA internal transcribed spacer (ITS) showed that these strains formed four divergent clusters: one close to Bradyrhizobium canariense and Bradyrhizobium lupini and three others separate from all the described species, representing three putative new Bradyrhizobium species. A phylogenetic analysis based on the nodC gene sequence affiliated the strains to either of the two symbiovars, genistearum or retamae.
Assuntos
Biodiversidade , Bradyrhizobium/classificação , Fabaceae/microbiologia , Filogenia , Nódulos Radiculares de Plantas/microbiologia , Argélia , Bradyrhizobium/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Genes Bacterianos/genética , Genoma Bacteriano/genética , Nodulação/genética , Análise de Sequência de DNA , Microbiologia do Solo , Simbiose/genéticaRESUMO
Fifty-one rhizobial strains isolated from root nodules of Cytisus villosus growing in Northeastern Algeria were characterized by genomic and phenotypic analyses. Isolates were grouped into sixteen different patterns by PCR-RAPD. The phylogenetic status of one representative isolate from each pattern was examined by multilocus sequence analyses of four housekeeping genes (16S rRNA, glnII, recA, and atpD) and one symbiotic gene (nodC). Analysis of 16S rRNA gene sequences showed that all the isolates belonged to the genus Bradyrhizobium. Phylogenetic analyses based on individual or concatenated genes glnII, recA, and atpD indicated that strains cluster in three distinct groups. Ten out of the sixteen strains grouped together with Bradyrhizobium japonicum, while a second group of four clustered with Bradyrhizobium canariense. The third group, represented by isolates CTS8 and CTS57, differed significantly from all other bradyrhizobia known to nodulate members of the Genisteae tribe. In contrast with core genes, sequences of the nodC symbiotic gene from all the examined strains form a homogeneous group within the genistearum symbiovar of Bradyrhizobium. All strains tested nodulated Lupinus angustifolius, Lupinus luteus, and Spartium junceum but not Glycine max. From these results, it is concluded that C. villosus CTS8 and CTS57 strains represent a new lineage within the Bradyrhizobium genus.