ABSTRACT: Areas affected by coal mining can be recovered by revegetation with leguminous plants associated with nitrogen-fixing bacteria. This study addressed the isolation and characterization of native nitrogen-fixing bacteria from coalmine wasteland under different vegetation restoration approaches using Macroptilium atropurpureum (DC) Urb and Vicia sativa L. as trap plants. The bacteria were characterized and identified on the basis of 16S rRNA sequences. Additionally, nitrogen-fixing strains were characterized for tolerance to high heavy metal and low pH levels, as well as for their effect on growth, nodulation, and symbiotic efficiency of M. atropurpureum and V. sativa. Soil samples were taken from the rhizosphere of eight areas, between 6 and 20 years under vegetation restoration, in the coal mining area of Candiota, RS-Brazil. The following properties were evaluated: colony characterization on solid “79” culture medium; pH (3.0-9.0) and heavy metal (Cr, Cd, Zn, Cu, and Ni) tolerance; partial sequencing of 16S rRNA gene; presence of nodA and nifH genes and symbiotic efficiency. A total of 115 isolates, i.e., 77 from M. atropurpureum and 38 from V. sativa, were obtained. The tolerance of these isolates is high for a wide range of pH levels and heavy metal contents, and 18 among them were selected for symbiotic efficiency and 16S rRNA sequencing. Inoculated with M. atropurpureum, the strains UFSM-B53, UFSM-B64, and UFSM-B74 had high symbiotic efficiency. The nitrogen-fixing bacteria were classified in the genera Rhizobium, Bradyrhizobium, and Burkholderia. The results indicate the potential of these native rhizospheric bacterial strains as inoculants and biofertilizers for legume species under pH and heavy metal stress in coal mining degraded areas in Southern Brazil.

ABSTRACT In Southern Brazil, agricultural soils are often treated over long periods with annual pig slurry (PS) applications, and their influence on soil nutrient dynamics should be monitored. This study aimed to evaluate the P forms accumulated in the soil and some biological and biochemical parameters of the P cycle after 19 successive PS applications over eight years, in a no-tillage (NT) system. Pig slurry was applied at rates of 0, 20, 40, and 80 m3 ha-1 per crop. Several P forms (available P, microbial P, phosphatase enzyme, total organic and inorganic P) in the 0.00-0.05 and 0.05-0.10 m soil layers were analyzed. Increases up to 5,523 % in available P, 308 % in microbial P, 104 % in phosphatase enzyme activity, 91 % in organic P, 251 % in inorganic P, and 19 % in Total-P were observed in the 0.00-0.05 m soil layer in response to the highest PS rate compared to the treatment with no PS. Results also indicated that P added via PS was preferentially accumulated in the inorganic fractions (83-98 %). The increase in organic P was limited by the increase in soil organic carbon. The increase in soil microbial biomass-P was more influenced by available P, while the increase in phosphatase enzyme activity was more related to the increase in organic P and C. Therefore, PS applied to the soil affected the chemical, biological, and biochemical components of the soil P cycle. Results indicate that even half of the PS rate normally used by farmers (20 m3 ha-1) poses a high risk of pollution of water bodies by runoff from these soils.

Práticas de manejo que visam reduzir a dependência de fertilizantes inorgânicos, como o uso de dejetos animais como fonte de P, podem contribuir para a sustentabilidade da agricultura. Embora informações sobre a biodisponibilidade e composição química do P presente nos dejetos utilizados como fertilizantes são importantes para o entendimento da sua dinâmica no solo, tais estudos ainda são escassos. Dessa forma, objetivou-se com o este trabalho avaliar o uso do fracionamento químico sequencial, da dessorção com resina de troca e da espectroscopia de ressonância magnética nuclear (31P-NMR) para caracterizar as formas de P presentes em três diferentes dejetos animais (ave, suíno e bovino). O fracionamento químico sequencial do P, a extração sucessiva com resina e a espectroscopia de 31P-RMN demonstraram que as formas de P presentes nos dejetos de aves, bovinos e suínos são principalmente inorgânicas e altamente biodisponíveis. O teor de P orgânico e recalcitrante foi desprezível e altamente dependente da relação Ca:P nos dejetos. Os resultados deste trabalho demonstram que o uso dos três diferentes métodos, quando aplicados conjuntamente, permite a caracterização completa do P presente nos dejetos animais.

Phosphate release kinetics from manures are of global interest because sustainable plant nutrition with phosphate will be a major concern in the future. Although information on the bioavailability and chemical composition of P present in manure used as fertilizer are important to understand its dynamics in the soil, such studies are still scarce. Therefore, P extraction was evaluated in this study by sequential chemical fractionation, desorption with anion-cation exchange resin and 31P nuclear magnetic resonance (31P-NMR) spectroscopy to assess the P forms in three different dry manure types (i.e. poultry, cattle and swine manure). All three methods showed that the P forms in poultry, cattle and swine dry manures are mostly inorganic and highly bioavailable. The estimated P pools showed that organic and recalcitrant P forms were negligible and highly dependent on the Ca:P ratio in manures. The results obtained here showed that the extraction of P with these three different methods allows a better understanding and complete characterization of the P pools present in the manures.