Diversity and Efficiency of Rhizobia Communities from Iron Mining Areas Using Cowpea as a Trap Plant

<div><p>ABSTRACT Mining is an important economic activity. However, its impact on environment must be accessed, mainly on relevant processes for their sustainability. The objective of this study was to evaluate the diversity and efficiency of symbiotic nitrogen fixing bacterial communities in soils under different types of vegetation in the Quadrilátero Ferrífero: ironstone outcrops, Atlantic Forest, neotropical savanna, and a rehabilitated area revegetated with grass. Suspensions of soil samples collected under each type of vegetation were made in a saline solution to capture rhizobia communities that were then inoculated on cowpea [Vigna unguiculata (L.) Walp.], which was used as a trap plant. The symbiotic efficiency of the communities was evaluated in a greenhouse experiment and the data obtained were correlated to the chemical and physical properties of the soils under each type of vegetation. At the end of the experiment, the bacteria present in the nodules were isolated to evaluate their diversity. The highest numbers of nodules occurred in the treatment inoculated with soil samples from rehabilitated area revegetated with grass and neotropical savanna vegetation, and the lowest numbers were observed in the treatment inoculated with soil samples from ironstone outcrops and Atlantic Forest. In relation to root dry matter, the treatment inoculated with soil samples from Neotropical savanah was superior to those inoculated with soil samples from the other areas; already, in relation to the shoot dry matter, no significant difference among the treatments was observed. The soil properties with the greatest influence on the microbial communities were Al3+ content, considered as high in the Atlantic Forest and neotropical savanna vegetation, as intermediate in the iron outcrops, and as very low in the rehabilitated area revegetated with grass; organic matter, considered as very high in the ironstone outcrops and neotropical savanna, as high in the Atlantic Forest, and as low in the rehabilitated area revegetated with grass; and the pH, with intermediate acidity level in the rehabilitated area revegetated with grass, high level of acidity in the iron outcrops and neotropical savanna, and very high acidity in the Atlantic Forest. After isolation of the nodules, 380 bacterial strains were obtained and separated into 27 groups by cultural characterization analysis. Genetic diversity was evaluated by the 16S rRNA gene partial sequencing of 156 strains, which identified some bacteria belonging to nitrogen-fixing Leguminosae nodulating bacterial genera (Rhizobium, Bradyrhizobium, Burkholderia, and Cupriavidus), some representative of associative bacteria (Bacillus, Paenibacillus, Herbaspirillum, Pseudomonas, and Agrobacterium), and other genera (Brevibacillus, Novosphingobium, Chitinophaga, Dyella, Acinetobacter, and Stenotrophomonas). The highest genetic diversity of bacteria was found in the rehabilitated area revegetated with grass indicated that it was effective in soil rehabilitation</p></div

Cupriavidus necator strains: zinc and cadmium tolerance and bioaccumulation

<div><p>ABSTRACT: Among soil microorganisms, the genus Cupriavidus has garnered particular scientific, economic and ecological interest because of its ability to fix nitrogen and tolerate high concentrations of metals. The aim of this study was to analyze four strains of Cupriavidus necator for their ability to tolerate and bioaccumulate cadmium and zinc. The tolerance of these strains to these metals was assessed in liquid culture medium containing different concentrations of Zn + Cd and in soil solutions and soils contaminated with multiple elements including Zn, Cd, Cu and Pb. The four strains showed high tolerance to Zn and Cd, both in culture medium and when inoculated into contaminated soil solutions or multi-element contaminated soil. The UFLA02-71 strain displayed the highest ability to bioaccumulate these metals. It was able to accumulate 93.76 µmol g cell−1 of Zn and 16.03 µmol g cell−1 of Cd when cultured in liquid medium with a total heavy metal concentration of 9,140 µmol L−1 (9,000 Zn + 140 Cd) and was able to accumulate 16.98 µmol g cell−1 of Cd in the soil solution. An increase in the pH of the culture medium resulting from the growth of the C. necator strains reduced the Zn2+ and Cd2+ ions in the medium and increased the concentrations of the ZnHPO4 and CdHPO4 species in solution. Thus, we concluded that they show great potential for use in the bioremediation of HM-contaminated areas.</p></div

Symbiotic Efficiency and Genotypic Characterization of Variants of Bradyrhizobium spp. in Commercial Inoculants for Soybeans

<div><p>ABSTRACT High yields obtained from soybean [Glycine max (L.) Merrill], and consequent profits, are related to efficient symbiosis of soybean with nitrogen fixing bacteria of the Bradyrhizobium genus. Inoculation with strains belonging to the Bradyrhizobium japonicum, B. elkanii, and B. diazofficiens species, approved by the Brazilian Ministry of Agriculture (MAPA), is a common practice for this crop. However, variations in the composition of inoculants can change the quality of the product. The aim of this study was to genetically identify and to evaluate the symbiotic efficiency of isolates with cultural characteristics different from those of strains approved by MAPA and which are present in two commercial inoculants for soybean. Isolates obtained from inoculants and the strains SEMIA 5079 (CPAC 15), SEMIA 5080 (CPAC 7), SEMIA 587 (BR 96), and SEMIA 5019 (BR 29), indicated in the formulation of these inoculants, were tested for symbiotic efficiency in soybeans (cultivar: COODETEC 5G 830 RR) in a greenhouse. Identification by partial sequencing of the 16S rRNA gene, together with BOX-PCR analysis, showed that variant isolates were not similar to the inoculant strains indicated in the commercial products, despite belonging to the same genus and/or species. No loss was observed in the nodulation ability of the isolates; however, significant differences were reported in the symbiotic efficiency of the isolates compared to the inoculant strains approved by MAPA. Nevertheless, the presence of morphological variants did not influence the efficiency of inoculant products.</p></div

Symbiosis of rhizobia with Gliricidia sepium and Clitoria fairchildiana in an Oxisol in the pre-Amazon region of Maranhão State

<div><p>ABSTRACT. Gliricidia (Gliricidia sepium) and sombreiro (Clitoria fairchildiana) have been recommended for agroforestry systems and reforestation of degraded areas due to their fast growth and symbiosis with rhizobia. However, little is known about native populations that nodulate these species. The objective of this study was to evaluate the phenotypic and genetic diversity of nitrogen-fixing bacteria isolated from nodules of gliricidia and sombreiro in alley cropping systems located in the pre-Amazon region of Maranhão State and to confirm their nodulation ability. Nodules were field collected from 20 plants of each species. The isolated strains were characterized morphologically, their 16S rRNA gene was partially sequenced, and their symbiotic ability was authenticated in siratro (Macroptilium atropurpureum). Despite being in the same climate and soil conditions, gliricidia and sombreiro are nodulated by different rhizobia genera, with Rhizobium predominant in gliricidia and Bradyrhizobium in sombreiro. Endophytic strains also colonized nodules in the field. Approximately 60% of Rhizobium strains did not nodulate siratro, whereas all Bradyrhizobium strains did. Native strains isolated from gliricidia nodules had low efficiency, and only four strains isolated from sombreiro nodules were efficient in siratro. These results highlight the importance of symbiotic relationships in the regulation of biological nitrogen fixation.</p></div

Arbuscular mycorrhizal fungal communities in an iron mining area and its surroundings: Inoculum potential, density, and diversity of spores related to soil properties

<div><p>ABSTRACT Arbuscular mycorrhizal fungi (AMF) interact symbiotically with most plant species, facilitating revegetation of areas under rehabilitation. The aim of this study was to evaluate the inoculum potential, density, and diversity of AMF spores in five environments, as well as the relation of species with soil properties. Soil samples were collected in five environments in a mining area and its surroundings in the Quadrilátero Ferrífero, Minas Gerais (Brazil): tailings piles in rehabilitation with grass, canga, Cerrado, native forest, and eucalyptus plantation; these samples were subjected to chemical and physical analyses. Spores were directly extracted from field samples and from trap cultures (TCs) established in two locations in the Southeast and South regions of Brazil for taxonomic identification of the species. Species richness, the Shannon diversity index (H’), and equitability were determined. Principal component analysis (PCA) was used to identify soil properties that most influenced AMF occurrence. Spore density showed no significant difference among the environments. A total of 59 AMF species were found. This is the first report of the occurrence of Acaulospora nivalis and Acaulospora alpina in Brazil. Higher H’ and species richness in the field were found in tailings piles and lower in canga. Canga showed higher inoculum potential. The development of TCs in two locations allowed a wider diversity of AMF species to be captured. Environments of the Quadrilátero Ferrífero are hotspots of AMF diversity, and the soil pH and exchangeable S and P contents are the properties that best explain the distribution of AMF species.</p></div

Tolerance to and Accumulation of Cadmium, Copper, and Zinc by Cupriavidus necator

<div><p>ABSTRACT Preliminary results of in vitro experiments with multicontaminated soils and solid media indicated that nodulating diazotrophic bacteria of the genus Cupriavidus are promising for the remediation of contaminated environments due to their symbiosis with legumes and metal tolerance. Thus, strains of Cupriavidus spp. (LMG 19424T, UFLA 01-659, UFLA 01-663, and UFLA 02-71) were tested for their ability to tolerate and bioaccumulate cadmium (Cd), copper (Cu), and zinc (Zn) in Luria-Bertani broth. Changes in the growth pattern of Cupriavidus strains in the presence or absence of heavy metals were analyzed by scanning electron microscopy and metal allocation by transmission electron microscopy, to clarify the mechanisms of bioremediation. Highest tolerance was detected for strain UFLA 01-659 (minimum inhibitory concentration of 5, 4.95, and 14.66 mmol L−1 of Cd, Cu, and Zn, respectively). Among the removal rates of the metals tested (9.0, 4.6, and 3.2 mg L−1 of Cd, Cu, and Zn, respectively), the bacterial activity was clearly highest for Cd. The efficiency of strain UFLA 01-659 in removing the heavy metals is associated with its high biomass production and/or higher contents of heavy metals adsorbed and absorbed in the biomass. In response to the presence of heavy metals in the liquid culture medium, the bacteria produced exopolysaccharides and small and aggregated cells. However, these responses varied according to the strains and heavy metals. Regarding allocation, all heavy metals were adsorbed on the cell wall and membrane, whereas complexation was observed intracellularly and only for Cu and Zn. These results indicate the possibility of using C. necator UFLA 01-659 for remediation in areas with very high Cd, Cu, and Zn contents.</p></div

Multiple linear regression and random forest to predict and map soil properties using data from portable X-ray fluorescence spectrometer (pXRF)

<div><p>ABSTRACT Determination of soil properties helps in the correct management of soil fertility. The portable X-ray fluorescence spectrometer (pXRF) has been recently adopted to determine total chemical element contents in soils, allowing soil property inferences. However, these studies are still scarce in Brazil and other countries. The objectives of this work were to predict soil properties using pXRF data, comparing stepwise multiple linear regression (SMLR) and random forest (RF) methods, as well as mapping and validating soil properties. 120 soil samples were collected at three depths and submitted to laboratory analyses. pXRF was used in the samples and total element contents were determined. From pXRF data, SMLR and RF were used to predict soil laboratory results, reflecting soil properties, and the models were validated. The best method was used to spatialize soil properties. Using SMLR, models had high values of R² (≥0.8), however the highest accuracy was obtained in RF modeling. Exchangeable Ca, Al, Mg, potential and effective cation exchange capacity, soil organic matter, pH, and base saturation had adequate adjustment and accurate predictions with RF. Eight out of the 10 soil properties predicted by RF using pXRF data had CaO as the most important variable helping predictions, followed by P2O5, Zn and Cr. Maps generated using RF from pXRF data had high accuracy for six soil properties, reaching R2 up to 0.83. pXRF in association with RF can be used to predict soil properties with high accuracy at low cost and time, besides providing variables aiding digital soil mapping.</p></div

Rhizobium strains selected from the Amazon region increase the yield of snap bean genotypes in protected cultivation

<div><p>ABSTRACT Although the use of inoculants containing rhizobia is encouraged, there are no official recommendations for inoculation of snap bean. In this respect, the aim of this study was to evaluate the agronomic performance of Rhizobium strains in symbiosis with snap bean cultivars with different growth habits and crop cycles. The experiment was carried out in pots with soil in a greenhouse in the spring-summer season in Lavras, state of Minas Gerais, Brazil. A completely randomized experimental design was used, with 4 replications, in a 4 × 7 factorial arrangement involving four snap bean cultivars and seven treatments – five strains of Rhizobium (UFLA 02-100, UFLA 02-127, UFLA 04-173, CIAT 899, and PRF 81), plus two controls without inoculation (with and without 500 mg N-NH4NO3∙dm–3). We conclude that: i) while genetic differences were observed among cultivars with respect to nodulating capacity, in all cases significant benefits were observed from BNF inoculation and ii) inoculation with selected strains of Rhizobium, as well as fertilization with mineral N, favors plant growtht, snap bean yields, and accumulation of N in the pods and, therefore, can fully substitute nitrogen fertilization in snap bean grown under protected cultivation.</p></div

Viability of liquid medium-inoculation of Rhizobium etli in planting furrows with common bean

<div><p>Abstract: The objective of this work was to evaluate the viability of liquid medium inoculation of Rhizobium etli in the planting furrow and to certify the efficiency of its strain UFLA 02-100 as a potential inoculant for common bean (Phaseolus vulgaris). The treatments consisted of three application rates of liquid inoculant applied in the planting furrows or in common bean seed, besides two controls without inoculation. The inoculant contained 109 cells of Rhizobium etli mL-1. Regardless of the application method, the yield obtained with the inoculation was equivalent to that of N from urea; however, the application in the furrows, at 0.6 L ha-1, is more advantageous due to the operational practicality and reduced costs.</p></div