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.

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.

Resumo: O objetivo deste trabalho foi avaliar a viabilidade da inoculação líquida de Rhizobium etli no sulco de semeadura e certificar a eficiência da estirpe UFLA 02-100 como inoculante potencial para o feijoeiro-comum (Phaseolus vulgaris). Os tratamentos consistiram de três doses do inoculante líquido aplicadas aos sulcos e uma dose aplicada às sementes de feijão-comum, além de dois controles sem inoculação. O inoculante continha 109células de Rhizobium etli mL-1. Independentemente do método de aplicação, a produtividade obtida com a inoculação foi equivalente à da aplicação de N da ureia, mas a aplicação ao sulco, com 0,6 L ha-1, é mais vantajosa pela praticidade operacional e redução de custos.

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.

RESUMO O objetivo deste trabalho foi avaliar a eficiência simbiótica de três novas estirpes de bactérias fixadoras de N2 em feijão-caupi cv. BRS Guariba como forrageira. Foram conduzidos dois experimentos em campo, em Balsas e São Raimundo das Mangabeiras, MA, em delineamento em blocos casualizados com sete tratamentos e quatro repetições, sendo: três novas estirpes eficientes em feijão-caupi (UFLA 3-153, UFLA 3-154 e UFLA 3-164), duas estirpes já aprovadas como inoculantes pelo MAPA (UFLA 3-84/SEMIA 6461 e INPA 3-11B/SEMIA 6462) e dois controles sem inoculação, com e sem N mineral. A eficiência simbiótica das estirpes foi determinada pelo número e massa seca de nódulos, produção de matéria seca da parte aérea, eficiência relativa, teor e acúmulo de proteína bruta e nitrogênio na parte aérea. As estirpes UFLA 03 -154 e UFLA 3-164 destacaram-se na eficiência simbiótica de N2 em Balsas e a UFLA 3-153 e UFLA 3-164 em São Raimundo das Mangabeiras. As estirpes UFLA 3-153, UFLA 3-154 e UFLA 3-164 são mais eficientes em fixar nitrogênio do que as estirpes aprovadas como inoculantes UFLA 3-84 e INPA 3-11B nas condições edafoclimáticas do sul do Maranhão. A UFLA 3-164 é a estirpe mais eficiente em fixar nitrogênio na região estudada.

ABSTRACT The objective of this study was to evaluate the symbiotic efficiency of three novel nitrogen-fixing bacterial strains with cowpea cv. BRS Guariba as forage. Two experiments were carried out in the field in Balsas and São Raimundo das Mangabeiras, MA, in a randomized block design with seven treatments and four replications: three novel strains efficient in symbiotic nitrogen fixation with cowpea (UFLA 3 -153, UFLA 3-154 and UFLA 3-164), two strains already approved as inoculants by MAPA (UFLA 3-84/SEMIA 6461 and INPA 3-11B/SEMIA 6462), and two controls without inoculation, with and without mineral N. Symbiotic efficiency of strains was determined by the number of nodules, nodules dry matter, shoot dry matter, relative efficiency, crude protein content and shoot N content. Strains UFLA 3-154 and UFLA 3-164 were the most efficient in Balsas, whereas UFLA 3-153 and UFLA 3-164 were the most efficient in São Raimundo das Mangabeiras. Strains UFLA 3-153, UFLA 3-154 and UFLA 3-164 were more efficient in symbiotic nitrogen fixation with cowpea than the strains already approved as inoculants UFLA 3-84 and INPA 3-11B in the climatic and edaphic conditions of southern Maranhão. UFLA 3-164 is the most efficient strain in this region.

RESUMO. Os objetivos do estudo foram avaliar a eficiência agronômica de estirpes aprovadas como inoculantes para feijão-caupi e de três novas estirpes sob seleção, bem como avaliar a influência da calagem sobre a simbiose e a produtividade. Dois experimentos de campo foram conduzidos utilizando feijão-caupi (BRS Guariba) de junho a setembro de 2013. No primeiro experimento, o delineamento experimental utilizado foi em blocos ao acaso, com quatro repetições e sete tratamentos (INPA03-11B, UFLA03-84, UFLA03-153, UFLA03-154, UFLA03-164 e dois controles). No segundo experimento, um fatorial 7 x 2 foi utilizado, sendo o primeiro fator composto pelos tratamentos acima e a calagem o segundo fator. Os parâmetros avaliados foram: matéria seca de nódulos, matéra seca de parte aérea, teor e acúmulo de N na parte aérea, número de vagens por planta, número de grãos por vagem, peso de 100 sementes, rendimento de grãos e teor e acúmulo de N no grão. As estirpes UFLA03-153 e UFLA03-164 foram mais eficientes para a inoculação do feijão-caupi do que as estirpes aprovadas como inoculantes, UFLA03-84 e INPA03-11B. Apesar de as estirpes e da cultivar testadas serem tolerantes à acidez do solo, a produtividade de feijão-caupi foi maior quando o solo recebeu calagem. As produções obtidas com os tratamentos de inoculação fertilizados foram bem acima da média do estado do Maranhão.

ABSTRACT. The objetives of the study were to evaluate the agronomic efficiency of strains approved as inoculants for cowpea and of three new strains under selection, as well as to evaluate the influence of liming on the symbiosis and productivity. Two field experiments were conducted using cowpea (BRS Guariba) from June to September 2013. In the first experiment, a completely randomized block experimental design was used, with four replicates and seven treatments (INPA03-11B, UFLA03-84, UFLA03-153, UFLA03-154, UFLA03-164 and two controls). In the second experiment, a 7 x 2 factorial design was used, with the above described treatments being the first factor and liming as the second factor. Parameters evaluated were: nodule dry matter, shoot dry matter, shoot N content and accumulation, number of pods per plant, number of seeds per pod, weight of 100 seeds, grain yield and grain N content and accumulation. Strains UFLA03-153 and UFLA03-164 were more efficient for cowpea inoculation than the strains approved as inoculants, UFLA03-84 and INPA03-11B. Although the tested rhizobia strains and cultivar BRS Guariba are tolerant to soil acidity, productivity washigher when soil was limed. Yields obtained with fertilized and inoculated treatments were well above Maranhão state average.

RESUMO. Objetivou-se avaliar os efeitos da aplicação foliar de molibdênio e do emprego de pequena dose de nitrogênio no plantio sobre a produção do feijoeiro quando se utiliza a inoculação das sementes com rizóbio. Foram conduzidos dois experimentos em campo, em delineamento de blocos ao acaso, esquema fatorial (3x2x2) + 1, com três repetições. Os tratamentos foram inoculação (estirpe CIAT 899T, estirpe UFLA 02-100 e ausência de inoculação), aplicação foliar de molibdênio (0 e 80 g ha-1) e de nitrogênio na semeadura (0 e 20 kg ha-1), mais tratamento adicional com N (40 na semeadura e 40 kg ha-1 na cobertura). A aplicação foliar de molibdênio não favoreceu a nodulação, mas seu efeito sobre o rendimento de grãos foi dependente das características químicas do solo, fornecendo maior produtividade em solo de Patos de Minas, com pH mais ácido e maiores teores de fósforo e matéria orgânica. Pequena dose de nitrogênio no plantio não reduziu a nodulação do feijoeiro inoculado com rizóbio, mas pode reduzir a emergência das sementes, com reflexo negativo na produtividade. Os rizóbios nativos proporcionaram nodulação, crescimento vegetal, acúmulo de nitrogênio na parte aérea e rendimento equivalentes aos tratamentos inoculados com as estirpes de rizóbio.

ABSTRACT. The aim of this study was to evaluate the effects of foliar application of molybdenum and small dose of nitrogen at planting on common bean production when using seed inoculation with rhizobia. Two field experiments were conducted, applying a randomized block design and a factorial arrangement (3x2x2) + 1, with three replicates. Treatments consisted of inoculation with Rhizobium spp. (strain CIAT 899T, strain UFLA 02-100, and uninoculated), foliar application of molybdenum (0 and 80 g ha-1), and fertilization with N-urea at planting (0 and 20 kg ha-1), with an additional N treatment of 40 applied at planting and 40 kg ha-1 applied as topdressing. The foliar application of molybdenum did not favor the nodulation, but its effect on yield was dependent on the chemical soil characteristics, providing highest yield in soil at Patos de Minas with pH more acid and with higher phosphorus and organic matter concentrations. Small dose of nitrogen applied at planting did not reduce nodulation of bean inoculated with rhizobia, but it can reduce the seeds emergence, thus negatively affecting yield. The native rhizobia promoted nodulation, the plant growth, shoot N accumulation and yield similar to the treatments that received inoculations of rhizobia strains.

ABSTRACT Management of biological nitrogen fixation in common bean still requires improvement. The objective of this study was to verify the compatibility of nitrogen fertilization with biological N2 fixation to increase common bean yield and profitability. Four field experiments were performed in four municipalities of Minas Gerais, Brazil, during the 2009 and 2010 winter crop season. The 2009 crop experiments were set up under a no-till system and the 2010 crop was conducted using conventional tillage. A randomized block experimental design was used with four replications and seven treatments combining application rates of mineral N (at sowing and/or topdressing) and seed inoculation with Rhizobium tropici strain CIAT899. Inoculation with 20 kg ha-1 N-urea at sowing and seed inoculation does not interfere with nodule dry matter and promotes yield comparable to that observed with 80 kg ha-1 N-urea with economic profitability in both no-till and conventional tillage systems. These results show the possibility of economic savings with respect to N fertilizers, but also a significant ecological contribution by avoiding problems associated with misuse of these fertilizers, such as eutrophication of waterways and denitrification.

ABSTRACT: Optimization of symbiosis between nitrogen-fixing bacteria and legumes has been extensively studied, seeking agricultural sustainability. To evaluate the symbiotic efficiency of nitrogen-fixing bacterial strains belonging to the Bradyrhizobium genus with pigeon pea (Cajanus cajan (L.) Millsp.) cv. Fava-Larga, experiments were conducted in Leonard jars (axenic conditions), pots with soil, and in the field. Ten strains were tested in Leonard jars, and three strains, in addition to BR 29, were selected according to their ability to promote the growth of pigeon pea, for further tests in pots with different soil types (Inceptsol and Oxisol) and in the field (Oxisol). Treatments were compared with strains BR 2003 and BR 2801 (approved as inoculants for pigeon pea), with a non-inoculated control with mineral N fertilization, and with another non-inoculated control (absolute control) with low mineral N concentration (Leonard jars) or without mineral N fertilization (soil). The efficiency of Bradyrhizobium strains in axenic conditions varies among strains, being higher when pigeon pea cv. Fava-Larga establishes symbiosis with the strains UFLA 03-320, UFLA 03-321, UFLA 04-212, BR 2801, and BR 2003. The soil type influences the symbiotic efficiency of Bradyrhizobium-pigeon pea in soil in the greenhouse, mainly in Inceptsol, in which strains UFLA 04-212, BR 2801, and BR 2003 increased N accumulation in the plant. The strain UFLA 03-320 increased shoot dry matter and N accumulation in the shoot equivalent to the mineral N treatment under field conditions. UFLA 03-320, BR 29, UFLA 03-321, and UFLA 04-212 promoted yields similar to those of the reference strain (BR 2801), and of the mineral N treatment with 70 kg ha-1 urea-N. These results confirm that pigeon pea establishes efficient symbiosis, which provides the N required for its growth. All strains, except for BR 2003, show potential for recommendation as inoculants for grain production. The strain UFLA 03-320 also shows potential for use in green manure crops.

Cowpea (Vigna unguiculata) cultivation in northern and northeastern Brazil provides an excellent source of nutrients and carbohydrates for the poor and underprivileged. Production surplus leads to its consumption in other regions of Brazil and also as an export commodity. Its capacity to establish relationships with atmospheric nitrogen-fixing bacteria is crucial to the reduction of production costs and the environmental impact of nitrogen fertilizers. This study assessed the symbiotic efficiency of new strains of symbiotic nitrogen-fixing bacteria with cowpea and their tolerance to pH and aluminum. Twenty-seven strains of bacteria from different soils were evaluated under axenic conditions. These strains were compared to the following inoculant strains: INPA03-11B, UFLA03-84 and BR3267 and two controls that were not inoculated (with and without mineral nitrogen). Six strains and the three strains approved as inoculants were selected to increase the dry weight production of the aerial part (DWAP) and were tested in pots with soil that had a high-density of nitrogen-fixing native rhizobia. In this experiment, three strains (UFLA03-164, UFLA03-153, and UFLA03-154) yielded higher DWAP values. These strains grow at pH levels of 5.0, 6.0, 6.8 and at high aluminum concentration levels, reaching 10(9) CFU mL-1. In particular UFLA03-84, UFLA03-153, and UFLA03-164 tolerate up to 20 mmol c dm-3 of Al+3. Inoculation with rhizobial strains, that had been carefully selected according to their ability to nodulate and fix N2, combined with their ability to compete in soils that are acidic and contain high levels of Al, is a cheaper and more sustainable alternative that can be made available to farmers than mineral fertilizers.

O objetivo deste trabalho foi avaliar a eficiência simbiótica de estirpes de bactérias fixadoras de nitrogênio do gênero Bradyrhizobium com guandu-anão. Os experimentos foram conduzidos em vasos de Leonard, em vasos com solo e em campo. Foram testadas 11 estirpes em vasos de Leonard, e as que apresentaram maior eficiência em promover o crescimento do guandu-anão foram avaliadas em vasos com solo (Latossolo Vermelho-Amarelo e Cambissolo) e em campo (Latossolo Vermelho-Amarelo). Em todos os experimentos, os tratamentos foram comparados a dois controles positivos (estirpes aprovadas como inoculantes para as cultivares de guandu-anão BR 2003 e BR 2801) e a duas testemunhas sem inoculação, uma com alta concentração de N mineral, e a outra, a depender do experimento, sem N mineral (solo) ou com baixa concentração de N (vasos de Leonard). Algumas estirpes proporcionaram crescimento vegetal semelhante ou superior às estirpes-referência e às testemunhas em vaso de Leonard. Em vasos com solo, o tipo de solo influenciou os tratamentos. No campo, não houve diferença entre os tratamentos, e as estirpes nativas promoveram bom crescimento. O guandu-anão é capaz de estabelecer associação simbiótica com bactérias fixadoras de N2, e a estirpe UFLA 03-320 apresenta potencial para ser recomendada para a cultura junto com a estirpe BR 2801.

The objective of this work was to evaluate the symbiotic efficiency of nitrogen-fixing bacterial strains, from the genus Bradyrhizobium, with dwarf pigeon pea. The experiments were carried out in Leonard jars, in pots with soil and in the field. In Leonard jars, 11 strains were tested, and the most effective in promoting pigeon pea growth were evaluated in pots with soil (Oxisol and Cambisol) and in the field (Oxisol). In all experiments, treatments were compared to two positive controls (strains approved as inoculants for pigeon pea cultivars, BR 2003 and BR 2801) and to two controls without inoculation, one with a high concentration of mineral N, and the other, depending on the experiment, without mineral N (soil) or with low N concentration (Leonard jars). Some strains have provided plant growth similar or superior to the reference strains and to the controls in Leonard jars. In the pot experiment with soil, the soil type influenced the treatments. In the field, there was no difference among treatments, and the native strains promoted a good plant growth. Dwarf pigeon pea is able to establish a symbiotic association with nitrogen-fixing bacteria, and the strain UFLA 03-320 has the potential to be recommended for this crop together with the BR 2801 strain.

O objetivo deste trabalho foi avaliar a tolerância de estirpes de Cupriavidus necator a zinco, cádmio, cobre e chumbo, além de determinar a eficiência simbiótica das estirpes mais tolerantes em associação a espécies leguminosas com potencial para revegetação. A tolerância foi testada em meio LB, suplementado com 2,5; 5,0; 7,5; 10; 12,5 e 15 mmol L-1 de ZnSO4.7H2O, CdSO4.8H2O, CuSO4.5H2O e PbCl2, respectivamente, em comparação ao controle sem adição de metal. Determinou-se a eficiência simbiótica das quatro estirpes de C. necator mais tolerantes aos metais avaliados (UFLA02-71, UFLA02-73, UFLA01-659 e UFLA01-663), as quais foram inoculadas nas espécies: Leucaena leucocephala, Enterolobium contortisiliquum, Acacia mangium, Mimosa caesalpiniifolia, M. pudica, M. pigra e M. acutistipula. Em vasos com solos, avaliaram-se L. leucocephala, M. pudica e M. caesalpiniifolia e as estirpes UFLA01-659 e UFLA02-71, selecionadas na avaliação de eficiência simbiótica. A estirpe UFLA02-71 proporcionou incrementos de matéria seca da parte aérea de 870% em M. caesalpiniifolia, enquanto que UFLA01-659 proporcionou 885% em M. pudica e 924% em L. leucocephala. As estirpes UFLA01-659 e UFLA02-71, além da alta tolerância a metais pesados, apresentaram eficiência em fixar nitrogênio, em simbiose com essas leguminosas, em solos com rizóbios nativos capazes de nodulá-las, e devem ser avaliadas quanto ao seu potencial de utilização em programas de recuperação de áreas degradadas.

The objective of this work was to evaluate the tolerance of Cupriavidus necator strains to zinc, cadmium, copper and lead, as well as to determine the symbiotic efficiency of the most tolerant ones in legume species suited for use in revegetation. Tolerance was evaluated in LB medium supplemented with 2.5, 5.0, 7.5, 10, 12.5 and 15 mmol L-1 of ZnSO4.7H2O, CdSO4.8H2O, CuSO4.5H2O and PbCl2, respectively, in comparison to a control without metal. The symbiotic efficiency of the four C. necator most metal-tolerant strains (UFLA02-71, UFLA02-73, UFLA01-659 and UFLA01-663) was determined, and these strains were inoculated in the species: Leucaena leucocephala, Enterolobium contortisiliquum, Acacia mangium, Mimosa caesalpiniifolia, M. pudica, M. pigra and M. acutistipula. Leucaena leucocephala, M. pudica and M. caesalpiniifolia and the strains UFLA02-71 and UFLA01-659, which presented the high symbiotic efficiency, were evaluated in pots with soil. UFLA02-71 provided increments of 870% in shoot dry matter of M. caesalpiniifolia, and UFLA01-659 provided 885% in M. pudica and 924% in L. leucocephala. These strains should be assessed for potential use in programs to restore degraded areas, since they showed high efficiency in nitrogen fixation and were competitive with indigenous rhizobia populations, besides being highly tolerant to heavy metals.

O objetivo deste trabalho foi avaliar o efeito do pH do meio de cultivo na eficiência simbiótica de estirpes de Rhizobium, em solo com e sem calagem. Foram realizados experimentos em casa de vegetação, com e sem calagem, e no campo, apenas com calagem. O delineamento experimental utilizado foi de blocos ao acaso com quatro repetições, e os tratamentos foram cinco estirpes de Rhizobium (UFLA 02-100, UFLA 02-68, UFLA 04-195, UFLA 04-202 e CIAT 899), cultivadas em meio de cultura 79 com diferentes valores de pH (5,0, 6,0 e 6,9), e testemunhas sem inoculação, com ou sem nitrogênio mineral. Foram avaliados número e massa de matéria seca de nódulos, massa de matéria seca da parte aérea, eficiência relativa, teor e acúmulo de nitrogênio na parte aérea e nos grãos, número de vagens por planta, número de grãos por vagem e rendimento e massa de 100 grãos. O valor de pH ideal para o meio de cultivo variou com as estirpes e com a variável analisada. Os tratamentos que receberam calagem foram superiores aos demais. No campo, independentemente do pH do inoculante, as populações nativas do solo e as estirpes introduzidas promoveram rendimento de grãos semelhante ao da testemunha com 70 kg ha-1 de N e ao da estirpe referência CIAT 899.

The objective of this study was to evaluate the effect of the culture medium pH in the symbiotic effectiveness of Rhizobium strains in soil, with and without liming. Assays were performed in greenhouse, with and without liming, and in the field, with liming. The experimental design was randomized blocks with four replicates, and treatments were five strains (UFLA 02-100, UFLA 02-68, UFLA 04-195, UFLA 04-202 and CIAT 899), grown in a 79 culture medium with different pH values (5.0, 6.0 and 6.9), and two controls without inoculation: with or without mineral nitrogen. Variables analyzed were number and weight of dry nodules, shoot dry matter, relative efficiency, nitrogen content and accumulation in shoots and grains, number of pods per plant, number of grains per pod, and yield and weight of 100 grains. The optimum pH value for the culture medium varied with the strains and the variable analyzed. Treatments with liming were superior to the others. In the field, regardless of the inoculant pH, the native soil populations and the introduced strains promoted grain yields similar to those of the control with 70 kg ha-1 of N and to those of the reference strain CIAT 899.