Microvirga vignae sp. nov., a root nodule symbiotic bacterium isolated from cowpea grown in semi-arid Brazil isolated from nodules of cowpea collected from a semi-arid region of Brazil showed 97 % similarity to sequences of recently described rhizobial species of the genus Microvirga. Phylogenetic analyses of four housekeeping genes (gyrB, recA, dnaK and rpoB), DNA-DNA relatedness and AFLP further indicated that these strains belong to a novel species within the genus Microvirga. Our data support the hypothesis that genes related to nitrogen fixation were obtained via horizontal gene transfer, as sequences of nifH genes were very similar to those found in members of the genera Rhizobium and Mesorhizobium, which are not immediate relatives of the genus Microvirga, as shown by 16S rRNA gene sequence analysis. Phenotypic traits, such as host range and carbon utilization, differentiate the novel strains from the most closely related species, Microvirga lotononidis, Microvirga zambiensis and Microvirga lupini. Therefore, these symbiotic nitrogen-fixing bacteria are proposed to be representatives of a novel species, for which the name Microvirga vignae sp. nov. is suggested. The type strain is BR3299 T (5HAMBI 3457 T ).

The biodiversity of rhizobium in soils of the São Francisco Valley is unknown and can be studied using cowpea as trap plants. The objective of this study was to verify the diversity of diazotrophic bacteria that nodulate cowpea in soils of the lower half of the São Francisco River Valley by morphological and genotypic characterization. Seven soil samples (A1, A2, A3, A4, C1, C2 and MC) were collected to capture bacteria associated to five cowpea cultivars (IPA 206, BRS Pujante, BRS Marataoã, Canapu Roxo, and Sempre Verde), in a 5x7 factorial design with three replications. Thirty days after plant emergence, the nodules were collected and the bacteria isolated and analyzed in relation to their growth characteristics in YMA medium. The 581 isolates were grouped in 49 morphologic groups. Of this total, 62.3 % formed colonies in up to three days, 33.4 % grew from the 6 th day on, and 4.3 % began to grow 4 to 5 days after incubation. Regarding the formation of acids and alkalis, 63 % acidified the medium, 12 % made it alkaline and 25 % maintained the medium at neutral pH. The highest diversity was observed in the A3 sample and in isolates associated with the cultivars Canapu Roxo and BRS Pujante. Thirty-eight representative isolates were chosen for the genotypic characterization, clustered in four groups based on the restriction analysis of 16s rDNA. This grouping was strongly correlated with the sampling site; 13 rhizobium isolates had an electrophoretic profile distinct from the standard rhizobium strains used in this study.

Clubroot, caused by Plasmodiophora brassicae, is a disease limiting the production of Brassica species. Its severity varies according to the density of the pathogen's resting spores and environmental conditions. Although the soil environment (including its physical, chemical and biological characteristics) is determinant in the development of the disease, little is known regarding these characteristics in tropical regions, such as Brazil. The objective of this study was to investigate the relationships between soil attributes, bacterial population composition and clubroot intensity in a tropical mountain agroecosystem. Soil and plant samples were collected from 17 cauliflower‐producing localities in Nova Friburgo, Brazil. The physical and chemical attributes, the bacterial population composition, the number of pathogen resting spores in the soil samples, and the clubroot severity were evaluated. Based on bacterial population composition, two distinct groups of soils were identified, which varied in attributes such as organic matter, potential acidity, pH and disease severity. The organic matter level was associated with differences in soil acidity, bacterial population composition and disease intensity, but there was not a cause and effect relationship between bacterial population composition and clubroot severity. Under the conditions of this agroecosystem, soils with increased organic matter presented higher acidity potential (H + Al), which was strongly correlated to clubroot severity. Thus, high acidity potential could be a key factor leading to clubroot development in mountain regions of Brazil.

The leguminous inoculation with nodule-inducing bacteria that perform biological nitrogen fixation is a good example of an “eco-friendly agricultural practice”. Bradyrhizobium strains BR 3267 and BR 3262 are recommended for cowpea (Vigna unguiculata) inoculation in Brazil and showed remarkable responses; nevertheless neither strain was characterized at species level, which is our goal in the present work using a polyphasic approach. The strains presented the typical phenotype of Bradyrhizobium with a slow growth and a white colony on yeast extract-mannitol medium. Strain BR 3267 was more versatile in its use of carbon sources compared to BR 3262. The fatty acid composition of BR 3267 was similar to the type strain of Bradyrhizobium yuanmingense; while BR 3262 was similar to Bradyrhizobium elkanii and Bradyrhizobium pachyrhizi. Phylogenetic analyses based on 16S rRNA and three housekeeping genes placed both strains within the genus Bradyrhizobium: strain BR 3267 was closest to B. yuanmingense and BR 3262 to B. pachyrhizi. Genome average nucleotide identity and DNA–DNA reassociation confirmed the genomic identification of B. yuanmingense BR 3267 and B. pachyrhizi BR 3262. The nodC and nifH gene analyses showed that strains BR 3267 and BR 3262 hold divergent symbiotic genes. In summary, the results indicate that cowpea can establish effective symbiosis with divergent bradyrhizobia isolated from Brazilian soils.

This study evaluated 26 pigeonpea rhizobial isolates according to their cultural characteristics, intrinsic antibiotic resistance, salt and temperature tolerance, carbon source utilization and amylolytic activity. The cultural characterization showed that the majority of them presented the ability to acidify the YMA. Among the 27 isolates evaluated, 25 were able to grow when incubated at 42° C and 11 showed tolerance to 3% (w/v) of NaCl in YMA medium. The patterns of carbon sources utilization was very diverse among the isolates. It was observed the capacity of three strains to metabolize all the carbon sources evaluated and a total of 42% of the bacterial isolates was able to grow in the culture medium supplemented with at least, six carbon sources. The carbon sources mannitol (control) and sucrose were metabilized by all isolates evaluated. The profile of intrinsic resistance to antibiotics showed that the isolates were mostly resistant to streptomycin and ampicillin, but susceptible to kanamycin and chloranphenicol. High amylolytic activity of, at least, four isolates was also demonstrated, especially for isolated 47.3b, which showed the highest enzymatic index. These results indicate the metabolic versatility of the pigeonpea rhizobia, and indicates the isolate 47.3b to further studies regarding the amylase production and characterization.

The Brazilian Northeastern dry forest (Caatinga) is one of the diversification centers of Mimosa species. We determined the characteristics of native rhizobia isolates from nodules of Mimosa tenuiflora and Mimosa paraibana grown in pots with soils collected under Caatinga vegetation and compared the restriction ribosomal DNA profiles of the isolates with those of 16 reference strains. All plants formed abundant indeterminate nodules and all nodule isolates formed fast growing colonies. No colony altered the medium to an alkaline reaction and most of them produced low or medium amounts of extracellular polysaccharides. White and creamy colonies predominated among the isolates but orange and green colonies were present. Differences among the isolates from the Mimosa species tested are indicated by the greater phenotypic diversity of those obtained from M. tenuiflora. The analysis of the 16S rDNA gene suggests that the isolates from M. tenuiflora and M. paraibana are closely related and closer to -rhizobia than to α-rhizobia. However, the similarity with all the tested -rhizobia reference strains was relatively low suggesting that the isolates may belong to different bacteria species.

As espécies de mucuna são muito utilizadas como adubos verdes, e poucas informações estão disponíveis a respeito dos rizóbios nativos capazes de nodulá-las. O objetivo deste trabalho foi avaliar a diversidade e a capacidade simbiótica de isolados bacterianos de nódulos de mucuna-cinza (Mucuna pruriens (L.) DC.) e mucuna-anã (Mucuna deeringiana (Bort.) Merr.). As bactérias foram isoladas de nódulos de mucunas cinza e anã cultivadas em vasos com solos de um sistema de produção agroecológica. Foram isoladas 160 bactérias, sendo 80 de mucuna-anã e 80 de mucuna-cinza, que foram autenticadas e selecionadas para avaliação da capacidade simbiótica. A diversidade dos isolados foi avaliada por meio das características culturais em meio de cultura YMA e da técnica de análise de restrição do produto de PCR do gene 16S rDNA. A inoculação de cinco isolados em mucuna-cinza e dois em mucuna-anã apresentou elevada biomassa da parte aérea. A maioria dos isolados apresentou crescimento rápido e acidificou o meio de cultura. A análise de restrição demonstrou que as bactérias isoladas apresentam baixa similaridade com estirpes de referência, sugerindo a existência de isolados pertencentes a novos grupos, capazes de nodular as mucunas anã e cinza.

Bacterias diazotróficas aisladas de arroz silvestre Oryza glumaepatula (Poaceae) en la Amazonia brasileña. The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg's medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH + bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fiftyone bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360bp nifH gene fragment. Among the 38 nifH + isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growthpromoting ability was confirmed for at least five isolates. For these bacteria, the root and shoot growing results showed higher increases when compared to those observed in plants inoculated with the evaluated reference strains. These results indicate that O. glumaepatula is colonized by a high diverse diazotrophic community in