Understanding native communities is a crucial step for the management of biological nitrogen fixation, since they may be either a source of efficient strains or a limiting factor when efficient strains need to be introduced. This work aimed to evaluate the density, diversity and efficiency of Leguminosae nodulating bacterial (LNB) communities and their component strains in soils under various land use systems (LUSs): pristine forest, agriculture, pasture, agroforestry, young secondary forest, and old secondary forest,. The LNB communities were trapped from these soils by using the promiscuous host siratro under controlled conditions. We also studied their relationships with physical and chemical attributes of the soil. Agroforestry and agriculture soil samples induced the highest number of nodules in siratro, while forest soil samples induced the lowest number of nodules. No relationship was found between LNB and Leguminosae species diversity in the LUSs. The soil chemical variables that were most related to differences in nodule number and shoot dry matter weight of plants inoculated with soil suspensions of the LUSs were, respectively: Ca 2+ , Mg 2+ , base saturation, exchangeable bases and Cu 2+ ; and pH, cation exchange capacity, B, Cu 2+ and clay. Although, LNB communities from all LUSs were efficient under controlled and similar conditions, they were found to be composed of strains with variable efficiency: inefficient, efficient, highly efficient and superior efficiency. Efficient strains occurred at the highest frequency in all LUSs. The isolated strains presented similar and new sequences that were phylogenetically related to well known LNB genera in α-and β-Proteobacteria. Unusual genera in these branches, as well as in other branches, which are probably endophytic bacteria, were also isolated from nodules. These data support siratro as a useful trap species to study the LNB biodiversity of diverse ecosystems in tropical soils. The fact that the highest diversity and nodulation were seen in managed systems such as agriculture and agroforestry suggests a high resilience of LNB communities to changes in land use after deforestation in a region where large forest areas are still preserved and can be a source of propagules.

The use of dark septate fungi (DSE) to promote plant growth can be beneficial to agriculture, and these organisms are important allies in the search for sustainable agriculture practices. This study investigates the contribution of dark septate fungi to the absorption of nutrients by rice plants and their ensuing growth. Four dark septate fungi isolates that were identified by Internal transcribed spacer phylogeny were inoculated in rice seeds (Cv. Piauí). The resulting root colonization was estimated and the kinetic parameters Vmax and Km were calculated from the nitrate contents of the nutrient solution. The macronutrient levels in the shoots, and the NO3−-N, NH4+-N, free amino-N and soluble sugars in the roots, sheathes and leaves were measured. The rice roots were significantly colonized by all of the fungi, but in particular, isolate A103 increased the fresh and dry biomass of the shoots and the number of tillers per plant, amino-N, and soluble sugars as well as the N, P, K, Mg and S contents in comparison with the control treatment. When inoculated with isolates A103 and A101, the plants presented lower Km values, indicating affinity increases for NO3−-N absorption. Therefore, the A103 Pleosporales fungus presented the highest potential for the promotion of rice plant growth, increasing the tillering and nutrients uptake, especially N (due to an enhanced affinity for N uptake) and P.

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Several processes that promote plant growth were investigated in endophytic and symbiotic bacteria isolated from cowpea and siratro nodules and also in bacterial strains recommended for the inoculation of cowpea beans. The processes verified in 31 strains were: antagonism against phytopathogenic fungi, free-living biological nitrogen fixation, solubilization of insoluble phosphates and indole acetic acid (IAA) production. The resistance to antibiotics was also assessed. Sequencing of the partial 16S rRNA gene was performed and the strains were identified as belonging to different genera. Eight strains, including some identified as Burkholderia fungorum, fixed nitrogen in the free-living state. Eighteen strains exhibited potential to solubilize calcium phosphate, and 13 strains could solubilize aluminum phosphate. High levels of IAA production were recorded with L-tryptophan addition for the strain UFLA04-321 (42.3 μg mL⁻¹). Strains highly efficient in symbiosis with cowpea bean, including strains already approved as inoculants showed the ability to perform other processes that promote plant growth. Besides, these strains exhibited resistance to several antibiotics. The ability of the nitrogen-fixing bacteria to perform other processes and their adaptation to environmental conditions add value to these strains, which could lead to improved inoculants for plant growth and environmental quality.

The legume species Sesbania virgata establishes a specific and efficient symbiosis with Azorhizobium doebereinerae. Previous studies have shown that A. doebereinerae occurrence correlates to the presence of S. virgata. This work aimed to evaluate the occurrence of A. doebereinerae and of other nitrogen-fixing Leguminosae-nodulating bacteria (NFLNB) in soil samples collected adjacent to and 10 m away from the stems of five S. virgata plants in pasture areas. Symbiotic characteristics of isolates from these NFLNB populations were also studied. S. virgata and the four promiscuous legume species Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris and Vigna unguiculata were inoculated with soil samples to trap A. doebereinerae and other NFLNB. NFLNB capable of inducing nodulation in at least one of these legumes were found in all samples. M. atropurpureum was the most promiscuous species, as it trapped the highest number of NFLNB cultural types from soil suspensions. The other species were less promiscuous in the following order: V. unguiculata, P. vulgaris, and L. leucocephala. Isolates of the promiscuous legumes were classified into seven cultural groups. One of these groups, isolated from all promiscuous species, showed fast-growth alkali-reaction in culture medium (like Azorhizobium); it was identified as Cupriavidus. This is the first report of symbiosis of Cupriavidus with Papilionoideae species. The symbiotic efficiency of promiscuous hosts with NFLNB varied, but it was always less than that of controls with mineral nitrogen or an inoculant strain. S. virgata was efficiently nodulated only by A. doebereinerae, which occurred mainly in samples collected close to the plant stem, corroborating a high stimulus by its host species. A high diversity of NFLNB occurs as saprophytes close to the S. virgata root system.

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Non-symbiotic diazotrophic bacteria are amongst the most important functional groups of soildwelling microorganisms. These bacteria contribute to plant growth predominantly through biological N 2 fixation. Here, we evaluated the density and diversity of non-symbiotic diazotrophic bacteria in soils taken from diverse land use systems (LUS) in Amazonia using nitrogen-free media. A total of 30 soil samples were collected from the following LUS: pristine forest, young secondary forest, old secondary forest, agroforestry, agriculture and pasture. Bacterial density was evaluated by the most probable number (MPN) method utilizing N-free semi-solid media with varied compositions (JNFb, NFb, LGI and Fam). Individual isolates were characterized by colony and cellular morphology as well as total protein profiles and nitrogenase activity. Isolate genotypes were determined by partial 16S rDNA sequences. No typical diazotrophic growth in the JNFb medium was observed. Bacterial densities in the NFb medium were higher in the agriculture and agroforestry soil samples. In LGI and Fam media, bacterial densities were highest in the pasture soil samples. Overall, 22 isolates with high phenotypic diversity were obtained. Eleven isolates exhibited nitrogenase activity. Sequences of 16S rDNA genes of 14 out of 19 isolates had similarities below 100 % with known nitrogen-fixing species. Isolates were identified as belonging to the Burkholderia, Enterobacter, Serratia, Klebsiella, and Bacillus genera. A higher number of isolates from pasture soil samples were isolated, with the majority of these belonging to the Burkholderia and Bacillus genera. Among the isolates, unknown sequences were obtained, possibly indicating new species. Taken together, these data demonstrate that Fam, NFb, and LGI semi-solid media allowed the growth of diazotrophic bacteria belonging to different phylogenetic lines.