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Phenotypic characterization and diversity of diazotrophic associative bacteria isolated from soils rehabilitated after bauxite mining

Grass species, which are frequently used for rehabilitation of degraded areas, can establish root associations with nitrogen fixing bacteria thus contributing to the ecosystem's sustainability. On the other hand, microbial diversity plays an important role in the resilience of biological processes, including N2 fixation. This study aimed at the phenotypic characterization of 72 diazotrophic Gram-negative bacteria. Samples were isolated from areas under different rehabilitation strategies after bauxite mining in Poços de Caldas, Minas Gerais, Brazil, and inoculated in NFb, Fam, and JNFb media. Type and reference strains of Herbaspirillum, Azospirillum and Burkholderia species were used for comparison as they are able to grow in such media. The similarity dendrogram based on seven cultural characteristics of the isolates in GNA medium presented a great diversity, as 50 groups were formed with 81 % similarity. NaCl tolerance in the potato/sucrose/acid malic medium varied from 0 to 50 g L-1 and allowed a separation of isolates and type strains into five groups. Cell diameters varied from 0.61 to 1.21 µm, and 13 isolates were not similar to the type strains. Fifteen groups with 75 % similarity were formed based on total proteins patterns obtained by polyacrylamide gel electrophoresis (SDS-PAGE). Neither was there any relationship among groups based on the different characteristics, nor between these and the areas of bacteria isolation. Fam and JNFb media detected the target species as well as other unidentified ones. The high phenotypic dissimilarity among isolates and type strains, mainly regarding total protein eletrophoresis profiles suggests that new species could be present within these populations.

Herbaspirillum; Azospirillum; Burkholderia; total cellular proteins; NFb; Fam; JNFb


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