Recovery of arsenic contaminated areas is a challenge society faces throughout the world. Revegetation associated with microbial activity can play an essential role in this process. This work investigated biological attributes in a gold mining area with different arsenic contents at different sites under two types of extant revegetation associated with cover layers of the soil: BS, Brachiaria sp. and Stizolobium sp., and LEGS, Acacia crassicarpa, A. holosericea, A. mangium, Sesbania virgata, Albizia lebbeck and Pseudosamanea guachapele. References were also evaluated, comprising the following three sites: B1, weathered sulfide substrate without revegetation; BM, barren material after gold extraction and PRNH (private reserve of natural heritage), an uncontaminated forest site near the mining area. The organic and microbial biomass carbon contents and substrate-induced respiration rates for these sites from highest to lowest were: PRNH > LEGS > BS > B1 and BM. These attributes were negatively correlated with soluble and total arsenic concentration in the soil. The sites that have undergone revegetation (LEGS and BS) had higher densities of bacteria, fungi, phosphate solubilizers and ammonium oxidizers than the sites without vegetation. Principal component analysis showed that the LEGS site grouped with PRNH, indicating that the use of leguminous species associated with an uncontaminated soil cover layer contributed to the improvement of the biological attributes. With the exception of acid phosphatase, all the biological attributes were indicators of soil recovery, particularly the following: microbial carbon, substrate-induced respiration, density of culturable bacteria, fungi and actinobacteria, phosphate solubilizers and metabolic quotient.

Arbuscular mycorrhizal fungi play an important role on nutrient supply to plants, specially P.However, the availability of inoculants for large-scale usage in agriculture is still limited because these organisms are obligatory symbionts. The use of symbiosis stimulants such as flavonoids can be an alternative to improve the beneficial effects of mycorrhiza for plant nutrition. The aim of this study was to evaluate the effect of the isoflavonoid biostimulant formononetin (7-hydroxy, 4'-methoxy isoflavone) in combination with three levels of phosphorus fertilization on mycorrhizal colonization, nodulation, and productivity of soybean, under field conditions. A 3 x 4 factorial scheme (levels of P: 0, 60 and 120 kg ha -1 P 2 O 5 and doses of formononetin: 0, 25, 50 and 100 g ha -1 ) was used with five replicates. The following parameters were quantified at full bloom: plant height, shoot dry weight, nodule number, nodule dry weight, mycorrhizal colonization, and shoot N and P concentrations. Productivity was also evaluated at the end of the crop cycle. Formononetin stimulated mycorrhizal colonization at lower levels of P (0 and 60 kg ha -1 ), with colonization increasing from 56 to 64%. When applied with 60 kg ha -1 P 2 O 5 , formononetin increased soybean productivity to values similar to those observed when 120 kg ha -1 de P 2 O 5 , was applied. At doses above 50 g ha -1 , formononetin applied to the seeds can reduce the need of P fertilization by 50%.

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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 19424 T , 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.

This study aimed to evaluate the effects of two rehabilitation systems in sites contaminated by Zn, Cu, Pb, and Cd on biological soil attributes [microbial biomass carbon (Cmic), basal and induced respiration, enzymatic activities, microorganism plate count, and bacterial and fungal community diversity and structure by denaturing gradient gel electrophoresis (DGGE)]. These systems (S1 and S2) consisted of excavation (trenching) and replacement of contaminated soil by uncontaminated soil in rows with Eucalyptus camaldulensis planting (S1-R and S2-R), free of understory vegetation (S1-BR), or completely covered by Brachiaria decumbens (S2-BR) in between rows. A contaminated, non-rehabilitated (NR) site and two contamination-free sites [Cerrado (C) and pasture (P)] were used as controls. Cmic, densities of bacteria and actinobacteria, and enzymatic activities (β-glucosidase, acid phosphatase, and urease) were significantly higher in the rehabilitated sites of system 2 (S2-R and S2-BR). However, even under high heavy metal contents (S1-R), the rehabilitation with eucalyptus was also effective. DGGE analysis revealed similarity in the diversity and structure of bacteria and fungi communities between rehabilitated sites and C site (uncontaminated). Principal component analysis showed clustering of rehabilitated sites (S2-R and S2-BR) with contamination-free sites, and S1-R was intermediate between the most and least contaminated sites, demonstrating that the soil replacement and revegetation improved the biological condition of the soil. The attributes that most explained these clustering were bacterial density, acid phosphatase, β-glucosidase, fungal and actinobacterial densities, Cmic, and induced respiration.

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Spore counts, species composition and richness of arbuscular mycorrhizal fungi, and soil glomalin contents were evaluated in a soil contaminated with Zn, Cu, Cd and Pb after rehabilitation by partial replacement of the contaminated soil with non-contaminated soil, and by Eucalyptus camaldulensis planting with and without Brachiaria decumbens sowing. These rehabilitation procedures were compared with soils from contaminated non-rehabilitated area and non-contaminated adjacent soils. Arbuscular mycorrhizal fungi communities attributes were assessed by direct field sampling, trap culture technique, and by glomalin contents estimate. Arbuscular mycorrhizal fungi was markedly favored by rehabilitation, and a total of 15 arbuscular mycorrhizal fungi morphotypes were detected in the studied area. Species from the Glomus and Acaulospora genera were the most common mycorrhizal fungi. Number of spores was increased by as much as 300-fold, and species richness almost doubled in areas rehabilitated by planting Eucalyptus in rows and sowing B. decumbens in inter-rows. Contents of heavy metals in the soil were negatively correlated with both species richness and glomalin contents. Introduction of B. decumbens together with Eucalyptus causes enrichment of arbuscular mycorrhizal fungi species and a more balanced community of arbuscular mycorrhizal fungi spores in contaminated soil.

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