Influence of common bean genotypes and rhizobia interaction for nodulation and nitrogen fixation / Influência da interação entre genótipos de feijoeiro e rizóbios na nodulação e fixação de nitrogênio

Bacteria from the rhizobia group are able to associate symbiotically with bean crop, forming nodules in the root, in which the biological nitrogen-fixing process occurs. However, the efficiency of this process has been low and it can be attributed to genetic and environmental factors. Thus, the objective of this study was to evaluate the capacity of nodulation of local varieties and commercial common bean cultivars inoculated with a Rhizobium tropici strain used in commercial inoculants and rhizobia isolates from common bean root nodules. The experiment was carried out in a factorial scheme (2x4), in a randomized block design with four replicates. It was tested two local varieties and two commercial cultivars, inoculated with the reference Rhizobium tropicistrain CIAT899 and the RBZ14 strain isolated from common bean nodules grown in soils of Southern Brazil, in adapted Leonard-type pots. The CIAT899 strain promoted either higher mass of viable nodules and higher nitrogen accumulation in the aerial part. The black group local variety showed better response than the cultivar TAA Dama for nodule viability, suggesting more efficiency for nodulation. The interaction between genotypes (local varieties and commercial cultivars) and bacteria showed the specificity of the complex symbiotic relationship of biological nitrogen fixation in common bean, requiring further studies of these interactions.(AU)

As bactérias do grupo rizóbios estão associadas simbioticamente à cultura do feijão, formando nódulos na raiz, nos quais ocorre o processo biológico de fixação do nitrogênio. No entanto, a eficiência desse processo tem sido baixa e pode ser atribuída a fatores genéticos e ambientais. Assim, este estudo teve como objetivo avaliar a capacidade de nodulação de variedades locais e cultivares comerciais de feijão comum inoculadas com uma estirpe de Rhizobium tropiciusada em inoculantes comerciais e isolados de nódulos da raiz do feijão comum. O experimento foi realizado em esquema fatorial (2x4), em delineamento de blocos ao acaso, com quatro repetições. Foram testadas duas variedades locais e duas cultivares comerciais, inoculadascom a estirpe de referência Rhizobium tropiciCIAT899 e o isolado RBZ14 obtido de nódulos de feijão cultivados em solos do sul do Brasil, em vasos adaptados do tipo Leonard. A estirpe CIAT899 promoveu maior massa de nódulos viáveis e maior acúmulo de nitrogênio na parte aérea. A variedade regional do grupo preto apresentou melhor resposta que a cultivar TAA Dama, quanto à viabilidade dos nódulos, sugerindo resposta mais eficiente à nodulação. A interação entre genótipos e bactérias mostrou a especificidade da complexa relação simbiótica da fixação biológica de nitrogênio no feijoeiro, sendo necessário aprofundar os estudos nessas interações.(AU)

Co-inoculation of rhizobia, azospirilla and cyanobacteria for increasing common bean production / Co-inoculação de rizobio, azospirillum e cianobactérias no aumento da produção de feijão comum

The combined inoculation of Rhizobium (R. tropici+R. freirei), Azospirillum brasilense, and Anabaena cylindrica, a diazotrophic cyanobacterium, is a technology that has not yet been tested and established in the production of the common bean (Phaseolus vulgaris). The inoculation may be a promising strategy for increasing crop productivity by combining the benefits of biological nitrogen fixation with the production of plant growth phytohormones. Therefore, the objective of this study was to evaluate the co-inoculation of Rhizobium, Azospirillum brasilense, and Anabaena cylindrica as an alternative method for optimizing the symbiotic performance and development of the common bean at greenhouse conditions. The treatments were as follows: (T1) control without N and inoculation, (T2) N addition (100kg N ha-¹), (T3) Riz (addition of R. tropici+R. freirei), (T4) Azo (Azospirillum brasilense addition), (T5)Ana (Anabaena cylindrical addition), (T6) Riz+Azo, (T7) Riz+Ana, (T8) Azo+Ana, (T9) Riz+Azo+Ana. We used a completely randomized experimental design with four replications. The co-inoculation of Riz+Azo+Ana promoted plant height, root length and volume, shoot dry matter, accumulated shoot N, number and dry matter of nodules at flowering, number of grains per pod, hundred seed weight, andgrain production of the common bean, contributing to increased yield per plant. We observed an increase in common bean grain yield ranging from 62 to 84% after double and triple co-inoculation of rhizobia with azospirilla and/or cyanobacteria, with the highest yield observed in the plants inoculated with Riz+Azo+Ana (84%), similar to those observed in plants after N addition. However, field experiments are necessary to elucidate the performances of the inoculated beneficial microorganisms.

A inoculação associada de Rhizobium, Azospirillum e uma cianobactéria diazotrófica Anabaena cylindrica é uma tecnologia que ainda não foi testada e estabelecida para o feijão comum (Phaseolus vulgaris) e pode ser uma estratégia promissora para aumentar a produtividade das culturas, combinando os benefícios da fixação biológica de nitrogênio com a produção de fitohormônios de crescimento vegetal. Portanto, o objetivo deste estudo foi avaliar a co-inoculação de Rizobium, Azospirilum brasilense e Anabaena cylindrica, como uma alternativa para otimizar o desempenho simbiótico e o desenvolvimento do feijoeiro, em casa de vegetação. Os tratamentos foram: T1-control, T2-N-mineral (100 kg N ha-¹), T3- Rhizobium (R. tropici+R. freirei), T4- A. brasilense, T5- A. cylindrica, T6- Rhizobium + A. brasilense, T7- Riz + Ana, T8- Azo + Ana, T9- Riz + Azo + Ana. O delineamento experimental foi inteiramente casualizado, com quatro repetições. A co-inoculação de Rhizobium (R.tropici+R. freirei) + Azo + Ana promoveu aumento do feijão (IPR Campos Gerais) em altura das plantas, comprimento e volume de raiz, massa seca da parte aérea, N acumulado da parte aérea, número e matéria seca dos nódulos no florescimento, número de grãos por vagem, peso de cem grãos e de grãos por planta, o que contribuiu para incremento da produção. Houve um aumento na produção de grãos de feijão com co-inoculação dupla e tripla de rizobia com azospirila e / ou cianobactérias, variando de 62a 84%, com o maior aumento de produção observado no tratamento inoculado com Riz + Azo + Ana(84%), semelhante ao tratamento com nitrogênio mineral. No entanto, experimentos de campo podem contribuir para elucidar o desempenho de microrganismos benéficos inoculados.

Co-inoculation of rhizobia, azospirilla and cyanobacteria for increasing common bean production / Co-inoculação de rizobio, azospirillum e cianobactérias no aumento da produção de feijão comum

The combined inoculation of Rhizobium (R. tropici+R. freirei), Azospirillum brasilense, and Anabaena cylindrica, a diazotrophic cyanobacterium, is a technology that has not yet been tested and established in the production of the common bean (Phaseolus vulgaris). The inoculation may be a promising strategy for increasing crop productivity by combining the benefits of biological nitrogen fixation with the production of plant growth phytohormones. Therefore, the objective of this study was to evaluate the co-inoculation of Rhizobium, Azospirillum brasilense, and Anabaena cylindrica as an alternative method for optimizing the symbiotic performance and development of the common bean at greenhouse conditions. The treatments were as follows: (T1) control without N and inoculation, (T2) N addition (100kg N ha-¹), (T3) Riz (addition of R. tropici+R. freirei), (T4) Azo (Azospirillum brasilense addition), (T5)Ana (Anabaena cylindrical addition), (T6) Riz+Azo, (T7) Riz+Ana, (T8) Azo+Ana, (T9) Riz+Azo+Ana. We used a completely randomized experimental design with four replications. The co-inoculation of Riz+Azo+Ana promoted plant height, root length and volume, shoot dry matter, accumulated shoot N, number and dry matter of nodules at flowering, number of grains per pod, hundred seed weight, andgrain production of the common bean, contributing to increased yield per plant. We observed an increase in common bean grain yield ranging from 62 to 84% after double and triple co-inoculation of rhizobia with azospirilla and/or cyanobacteria, with the highest yield observed in the plants inoculated with Riz+Azo+Ana (84%), similar to those observed in plants after N addition. However, field experiments are necessary to elucidate the performances of the inoculated beneficial microorganisms.(AU)

A inoculação associada de Rhizobium, Azospirillum e uma cianobactéria diazotrófica Anabaena cylindrica é uma tecnologia que ainda não foi testada e estabelecida para o feijão comum (Phaseolus vulgaris) e pode ser uma estratégia promissora para aumentar a produtividade das culturas, combinando os benefícios da fixação biológica de nitrogênio com a produção de fitohormônios de crescimento vegetal. Portanto, o objetivo deste estudo foi avaliar a co-inoculação de Rizobium, Azospirilum brasilense e Anabaena cylindrica, como uma alternativa para otimizar o desempenho simbiótico e o desenvolvimento do feijoeiro, em casa de vegetação. Os tratamentos foram: T1-control, T2-N-mineral (100 kg N ha-¹), T3- Rhizobium (R. tropici+R. freirei), T4- A. brasilense, T5- A. cylindrica, T6- Rhizobium + A. brasilense, T7- Riz + Ana, T8- Azo + Ana, T9- Riz + Azo + Ana. O delineamento experimental foi inteiramente casualizado, com quatro repetições. A co-inoculação de Rhizobium (R.tropici+R. freirei) + Azo + Ana promoveu aumento do feijão (IPR Campos Gerais) em altura das plantas, comprimento e volume de raiz, massa seca da parte aérea, N acumulado da parte aérea, número e matéria seca dos nódulos no florescimento, número de grãos por vagem, peso de cem grãos e de grãos por planta, o que contribuiu para incremento da produção. Houve um aumento na produção de grãos de feijão com co-inoculação dupla e tripla de rizobia com azospirila e / ou cianobactérias, variando de 62a 84%, com o maior aumento de produção observado no tratamento inoculado com Riz + Azo + Ana(84%), semelhante ao tratamento com nitrogênio mineral. No entanto, experimentos de campo podem contribuir para elucidar o desempenho de microrganismos benéficos inoculados.(AU)

Changes induced by co-inoculation in nitrogencarbon metabolism in cowpea under salinity stress

To mitigate the deleterious effects of abiotic stress, the use of plant growth-promoting bacteria along with diazotrophic bacteria has been increasing. The objectives of this study were to investigate the key enzymes related to nitrogen and carbon metabolism in the biological nitrogen fixation process and to elucidate the activities of these enzymes by the synergistic interaction between Bradyrhizobium and plant growth-promoting bacteria in the absence and presence of salt stress. Cowpea plants were cultivated under axenic conditions, inoculated with Bradyrhizobium and co-inoculated with Bradyrhizobium sp. and Actinomadura sp., Bradyrhizobium sp. and Bacillus sp., Bradyrhizobium sp. and Paenibacillus graminis, and Bradyrhizobium sp. and Streptomycessp.; the plants were also maintained in the absence (control) and presence of salt stress (50 mmolL-1 NaCl). Salinity reduced the amino acids, free ammonia, ureides, proteins and total nitrogen content in nodules and increased the levels of sucrose and soluble sugars. The co-inoculations responded differently to the activity of glutamine synthetase enzymes under salt stress, as well as glutamate synthase, glutamate dehydrogenase aminating, and acid invertase in the control and salt stress. Considering the development conditions of this experiment, co-inoculation with Bradyrhizobium sp. and Bacillus sp. in cowpea provided better symbiotic performance, mitigating the deleterious effects of salt stress.(AU)

Genomic identification and characterization of the elite strains Bradyrhizobium yuanmingense BR 3267 and Bradyrhizobium pachyrhizi BR 3262 recommended for cowpea inoculation in Brazil

The leguminous inoculation with nodule-inducing bacteria that perform biological nitrogen fixation is a good example of an eco-friendly agricultural practice. Bradyrhizobium strains BR 3267 and BR 3262 are recommended for cowpea (Vigna unguiculata) inoculation in Brazil and showed remarkable responses; nevertheless neither strain was characterized at species level, which is our goal in the present work using a polyphasic approach. The strains presented the typical phenotype of Bradyrhizobium with a slow growth and a white colony on yeast extract-mannitol medium. Strain BR 3267 was more versatile in its use of carbon sources compared to BR 3262. The fatty acid composition of BR 3267 was similar to the type strain of Bradyrhizobium yuanmingense; while BR 3262 was similar to Bradyrhizobium elkanii and Bradyrhizobium pachyrhizi. Phylogenetic analyses based on 16S rRNA and three housekeeping genes placed both strains within the genus Bradyrhizobium: strain BR 3267 was closest to B. yuanmingense and BR 3262 to B. pachyrhizi. Genome average nucleotide identity and DNADNA reassociation confirmed the genomic identification of B. yuanmingense BR 3267 and B. pachyrhizi BR 3262. The nodC and nifH gene analyses showed that strains BR 3267 and BR 3262 hold divergent symbiotic genes. In summary, the results indicate that cowpea can establish effective symbiosis with divergent bradyrhizobia isolated from Brazilian soils.(AU)

Diferentes inoculantes e formas de inoculação e sua influência sobre os componentes de produção e teor de nitrogênio da cultura da soja / Different inoculants and forms of inoculation and its influence on the components of production and the content of nitrogen in soybean

A demanda de nitrogênio, durante o ciclo da cultura soja, é suprida pela associação simbiótica entre as raízes da soja e as bactérias do gênero Bradyrhizobium. Além disso, está disponível para o produtor a possibilidade de seutilizar bactérias promotoras do crescimento de plantas do gênero Azospirillum, que, em coinoculação, podem trazer benefícios para a nodulação da soja e na eficiência de absorção de nutrientes. O objetivo do presente trabalho foi o de avaliar a eficiênciadas bactérias Bradyrhizobiume Azospirillum brasilense, de forma isolada e mista, em diferentes formas de aplicação sobre os componentes de rendimento, produtividade e teor de nitrogênio da soja. O experimento foi conduzido através do delineamento em blocos casualizados com quatro tratamentos e cinco repetições, sendo eles: T1: Bradyrhizobium aplicado no sulco; T2: Bradyrhizobiumaplicado na semente; T3: Bradyrhizobium+ Azospirillum brasilenseno sulco; T4: Bradyrhizobium japonicum+ Azospirillum brasilenseaplicado via tratamento de sementes. A cultivar utilizada no trabalho foi a Nidera 5909 com 50 cm entre linhas. Avaliaram-se a altura de plantas, diâmetro do caule, número de nós reprodutivos, número de vagens por planta, número de grãos por vagem, massa de mil grãos, produtividade e teor de nitrogênio na folha. Somente para o teor de nitrogênio, a aplicação no sulco foi superior a inoculação e coinoculação na semente, a coinoculação no sulco apresentou a maior massa de mil grãos. A associação das duas bactérias proporcionou as maiores produtividades para as condições testadas.(AU)

The nitrogen demand during the soybean cycle is supplied by the symbiotic association between soybean roots and Bradyrhizobium bacteria. In addition, it is available to the grower the possibility of using bacteria that promote the growth of plants of the genus Azospirillum, which in coinoculation can bring benefits to soybean nodulation and nutrientabsorption efficiency. The objective of the present work was to evaluate the efficiency of the Bradyrhizobiumand Azospirillum brasilensebacteria, isolated and mixed in different forms of application on the components of yield, yield and nitrogen contentof soybean. The experiment was conducted in a randomized block design with four treatments and five replications: T1: Bradyrhizobiumapplied in the furrow; T 2: Bradyrhizobiumapplied to the seed; T 3: Bradyrhizobium+ Azospirillum brasilensein the furrow; T 4: Bradyrhizobium japonicum+ Azospirillum brasilenseapplied via seed treatment with 5 repetitions, using the cultivar Nidera 5909 with 50cm between rows. The variable responses evaluated were: Plant height, stem diameter, number of reproductive nodes, number of pods per plant, number of grains per pod, weight of thousand grains, yield and nitrogen content in the leaf. Only for the nitrogen content the application in the furrow was superior to the inoculation and co-inoculation in the seed, the co-inoculation in the furrow had the highest weight for a thousand grains. The combination of both bacteria gave the highest yields for the conditions tested.(AU)

Phenotypic, genetic and symbiotic characterization of Erythrina velutina rhizobia from Caatinga dry forest

Erythrina velutina ("mulungu") is a legume tree from Caatinga that associates with rhizobia but the diversity and symbiotic ability of "mulungu" rhizobia are poorly understood. The aim of this study was to characterize "mulungu" rhizobia from Caatinga. Bacteria were obteined from Serra Talhada and Caruaru in Caatinga under natural regeneration. The bacteria were evaluated to the amplification of nifH and nodC and to metabolic characteristics. Ten selected bacteria identified by 16S rRNA sequences. They were tested in vitro to NaCl and temperature tolerance, auxin production and calcium phosphate solubilization. The symbiotic ability were assessed in an greenhouse experiment. A total of 32 bacteria were obtained and 17 amplified both symbiotic genes. The bacteria showed a high variable metabolic profile. Bradyrhizobium (6), Rhizobium (3) and Paraburkholderia (1) were identified, differing from their geographic origin. The isolates grew up to 45 °C to 0.51 mol L-1 of NaCl. Bacteria which produced more auxin in the medium with l-tryptophan and two Rhizobium and one Bradyrhizobium were phosphate solubilizers. All bacteria nodulated and ESA 90 (Rhizobium sp.) plus ESA 96 (Paraburkholderia sp.) were more efficient symbiotically. Diverse and efficient rhizobia inhabit the soils of Caatinga dry forests, with the bacterial differentiation by the sampling sites.(AU)

Diferentes inoculantes e formas de inoculação e sua influência sobre os componentes de produção e teor de nitrogênio da cultura da soja / Different inoculants and forms of inoculation and its influence on the components of production and the content of nitrogen in soybean

A demanda de nitrogênio, durante o ciclo da cultura soja, é suprida pela associação simbiótica entre as raízes da soja e as bactérias do gênero Bradyrhizobium. Além disso, está disponível para o produtor a possibilidade de se utilizar bactérias promotoras do crescimento de plantas do gênero Azospirillum, que, em coinoculação, podem trazer benefícios para a nodulação da soja e na eficiência de absorção de nutrientes. O objetivo do presente trabalho foi o de avaliar a eficiência das bactérias Bradyrhizobium e Azospirillum brasilense, de forma isolada e mista, em diferentes formas de aplicação sobre os componentes de rendimento, produtividade e teor de nitrogênio da soja. O experimento foi conduzido através do delineamento em blocos casualizados com quatro tratamentos e cinco repetições, sendo eles: T1: Bradyrhizobium aplicado no sulco; T2: Bradyrhizobium aplicado na semente; T3: Bradyrhizobium + Azospirillum brasilense no sulco; T4: Bradyrhizobium japonicum + Azospirillum brasilense aplicado via tratamento de sementes. A cultivar utilizada no trabalho foi a Nidera 5909 com 50 cm entre linhas. Avaliaram-se a altura de plantas, diâmetro do caule, número de nós reprodutivos, número de vagens por planta, número de grãos por vagem, massa de mil grãos, produtividade e teor de nitrogênio na folha. Somente para o teor de nitrogênio, a aplicação no sulco foi superior a inoculação e coinoculação na semente, a coinoculação no sulco apresentou a maior massa de mil grãos. A associação das duas bactérias proporcionou as maiores produtividades para as condições testadas.(AU)

The nitrogen demand during the soybean cycle is supplied by the symbiotic association between soybean roots and Bradyrhizobium bacteria. In addition, it is available to the grower the possibility of using bacteria that promote the growth of plants of the genus Azospirillum, which in coinoculation can bring benefits to soybean nodulation and nutrient absorption efficiency. The objective of the present work was to evaluate the efficiency of the Bradyrhizobium and Azospirillum brasilense bacteria, isolated and mixed in different forms of application on the components of yield, yield and nitrogen content of soybean. The experiment was conducted in a randomized block design with four treatments and five replications: T1: Bradyrhizobium applied in the furrow; T 2: Bradyrhizobium applied to the seed; T 3: Bradyrhizobium + Azospirillum brasilense in the furrow; T 4: Bradyrhizobium japonicum + Azospirillum brasilense applied via seed treatment with 5 repetitions, using the cultivar Nidera 5909 with 50cm between rows. The variable responses evaluated were: Plant height, stem diameter, number of reproductive nodes, number of pods per plant, number of grains per pod, weight of thousand grains, yield and nitrogen content in the leaf. Only for the nitrogen content the application in the furrow was superior to the inoculation and co-inoculation in the seed, the co-inoculation in the furrow had the highest weight for a thousand grains. The combination of both bacteria gave the highest yields for the conditions tested.(AU)

Genome sequence of Bradyrhizobium embrapense strain CNPSo 2833T, isolated from a root nodule of Desmodium heterocarpon

Bradyrhizobium embrapense CNPSo 2833T is a nitrogen-fixing symbiont of the legume pasture Desmodium. Its draft genome contains 8,267,832 bp and 7876 CDSs. The symbiotic island includes nodulation and nitrogen fixation genes resembling the operon organization of B. japonicum. Several CDSs related to secretion proteins and stress tolerance were also identified.(AU)

Agronomic efficiency of Rhizobium strains from the Amazon region in common bean / Eficiência agronômica de estirpes de Rhizobium isoladas da Amazônia sobre o feijoeiro-comum

To maximize the contribution of biological nitrogen fixation in common bean, it is necessary to use bacterial strains that are more adapted, competitive, and efficient in the symbiotic process. In this regard, the aim of this study was to evaluate the agronomic efficiency (AE) of three bacterial strains isolated from acid soils with high Al content from the Amazon region in an Argissolo Vermelho Distrófico típico soil (Typic Rhodustults - USDA Classification) from the municipality of Formiga, MG, Brazil. We compared their AE to that of the reference strain CIAT 899T and of two controls without inoculation (one without and another with 80 kg ha-1 of N-urea). The results indicated that inoculation with the strains UFLA 02-100 and UFLA 02-127 provides grain yield equivalent to inoculation with the reference strain and to the control with mineral N. Thus, both have potential for recommendation as inoculants for common bean.

Para maximizar a contribuição da fixação biológica de nitrogênio no feijoeiro-comum é necessária a utilização de estirpes de bactérias mais adaptadas, competitivas e eficientes no processo simbiótico. Nesse sentido, objetivou-se avaliar, em um Argissolo Vermelho Distrófico típico do município de Formiga-MG, a eficiência agronômica (EA) de três estirpes isoladas de solos ácidos e com alto teor de Al da Amazônia e comparar suas EA à da estirpe referência CIAT 899T e à de dois controles sem inoculação (um sem e outro com 80 kg ha-1 of N-ureia). Os resultados indicaram que a inoculação com as estirpes UFLA 02-100 e UFLA 02-127 propicia rendimento de grãos equivalente ao da estirpe referência e ao do controle com N mineral e que por isso, ambas têm potencial para ser recomendadas como inoculantes para o feijoeiro-comum.

Agronomic efficiency of Rhizobium strains from the Amazon region in common bean / Eficiência agronômica de estirpes de Rhizobium isoladas da Amazônia sobre o feijoeiro-comum

To maximize the contribution of biological nitrogen fixation in common bean, it is necessary to use bacterial strains that are more adapted, competitive, and efficient in the symbiotic process. In this regard, the aim of this study was to evaluate the agronomic efficiency (AE) of three bacterial strains isolated from acid soils with high Al content from the Amazon region in an Argissolo Vermelho Distrófico típico soil (Typic Rhodustults - USDA Classification) from the municipality of Formiga, MG, Brazil. We compared their AE to that of the reference strain CIAT 899T and of two controls without inoculation (one without and another with 80 kg ha-1 of N-urea). The results indicated that inoculation with the strains UFLA 02-100 and UFLA 02-127 provides grain yield equivalent to inoculation with the reference strain and to the control with mineral N. Thus, both have potential for recommendation as inoculants for common bean.(AU)

Para maximizar a contribuição da fixação biológica de nitrogênio no feijoeiro-comum é necessária a utilização de estirpes de bactérias mais adaptadas, competitivas e eficientes no processo simbiótico. Nesse sentido, objetivou-se avaliar, em um Argissolo Vermelho Distrófico típico do município de Formiga-MG, a eficiência agronômica (EA) de três estirpes isoladas de solos ácidos e com alto teor de Al da Amazônia e comparar suas EA à da estirpe referência CIAT 899T e à de dois controles sem inoculação (um sem e outro com 80 kg ha-1 of N-ureia). Os resultados indicaram que a inoculação com as estirpes UFLA 02-100 e UFLA 02-127 propicia rendimento de grãos equivalente ao da estirpe referência e ao do controle com N mineral e que por isso, ambas têm potencial para ser recomendadas como inoculantes para o feijoeiro-comum.(AU)

Selection of efficient rhizobial symbionts for Cratylia argentea in the cerrado biome / Seleção de rizóbios eficientes na simbiose com Cratylia argentea no bioma cerrado

Cratylia argentea is a leguminous shrub native to the cerrado, which has great potential for forage production and recovery of degraded areas. This study aimed to isolate, characterize, and select efficient rhizobial strains in symbiosis with Cratylia argentea . Rhizobacteria were isolated from the nodules of 12-month-old plants and cultivated in pots containing cerrado soil. Twenty-five bacterial strains were obtained, which displayed extensive variability with respect to morphological and symbiotic characteristics. Cratylia argentea seeds were planted in pots containing 5kg of cerrado soil and maintained in the greenhouse. The treatments consisted of 25 rhizobial isolates, two controls (without nitrogen and without inoculation), with or without nitrogen fertilization (5mgN·plant-1·week-1), and four replications. Plants were cultivated for 150 days after planting seeds to evaluate nodule number, nodule dry weight, shoot and root dry weight, shoot and root N content, and relative and symbiotic efficiency. Thirteen isolates improved shoot dry weight (up to 65.8%) and shoot nitrogen concentration (up to 76%) compared with those of control treatments. Two isolates, 4 (CR42) and 22 (CR52), conferred higher symbiotic efficiency values of approximately 20%. Therefore, these two rhizobial isolates displayed the highest potential as beneficial inoculants to optimize the symbiotic efficiency for Cratylia and to increase the incorporation of nutrients and biomass into the productive system in the cerrado.(AU)

Cratylia argentea é uma leguminosa arbustiva nativa do cerrado, com alto potencial para produção de forragem e recuperação de áreas degradadas. Este trabalho teve como objetivo isolar, caracterizar e selecionar estirpes de rizóbios eficientes na simbiose com C. argentea . As bactérias foram isoladas de nódulos de plantas com 12 meses de idade, cultivadas em solo de cerrado, em vasos. Foram obtidas 25 estirpes, que apresentaram alta variabilidade em relação às características morfológicas e simbióticas. Sementes de C. argentea foram plantadas em potes contendo 5kg de solo de cerrado, e mantidos em casa de vegetação. Os tratamentos consistiram em 25 isolados, controle (sem N e sem inoculação) e adubação nitrogenada (5mgN·planta-1·semana-1), em quatro repetições. As plantas foram colhidas 150 dias após o plantio para avaliar o número e a massa seca de nódulos, a massa seca e concentração de nitrogênio da parte área e das raízes. Foram calculadas as eficiências relativas e simbióticas. Treze estirpes se destacaram quanto ao acúmulo de massa seca (até 65,8%) e acúmulo de nitrogênio (até 76%), da parte aérea, em relação ao tratamento controle. Dentre estas, as estirpes 4 e 22 apresentaram maiores eficiências simbióticas (em torno de 20%). Portanto, estas estirpes apresentaram potencial para formulação de inoculantes, visando otimizar o processo simbiótico rizóbio- Cratylia e a incorporação de biomassa e nutrientes em sistema produtivos no cerrado.(AU)

Draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite strain recommended for cowpea inoculation in Brazil

The strain BR 3267 is a nitrogen-fixing symbiotic bacteria isolated from soil of semi-arid area of Brazilian Northeast using cowpea as the trap plant. This strain is used as commercial inoculant for cowpea and presents high efficient in nitrogen fixation as consequence of its adaptation potential to semi-arid conditions. We report here the draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite bacterium used as inoculant for cowpea. Whole genome sequencing of BR 3267 using Illumina MiSeq sequencing technology has 55 scaffolds with a total genome size of 7,904,309 bp and C+G 63%. Annotation was added by the RAST prokaryotic genome annotation service and has shown 7314 coding sequences and 52 RNA genes.

Draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite strain recommended for cowpea inoculation in Brazil

The strain BR 3267 is a nitrogen-fixing symbiotic bacteria isolated from soil of semi-arid area of Brazilian Northeast using cowpea as the trap plant. This strain is used as commercial inoculant for cowpea and presents high efficient in nitrogen fixation as consequence of its adaptation potential to semi-arid conditions. We report here the draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite bacterium used as inoculant for cowpea. Whole genome sequencing of BR 3267 using Illumina MiSeq sequencing technology has 55 scaffolds with a total genome size of 7,904,309 bp and C+G 63%. Annotation was added by the RAST prokaryotic genome annotation service and has shown 7314 coding sequences and 52 RNA genes.(AU)

Nursery growth and rhizobia symbiosis of scandent Leguminosae species native to the Amazon region / Crescimento e simbiose com rizóbios em casa de vegetação de espécies de Leguminosae escandentes nativas da região Amazônica

A great number of species and individuals of scandent legumes establishing symbiosis with nitrogen fixing bacteria occurs in the Amazon Forest. These symbiosis probably play an important role in contributing to nitrogen incorporation in this ecossystem. The objectives of this study were to evaluate the growth of eight species of scandent legumes in five nursery substrates; to compare nodulation with rhizobia strains introduced or native to these substrates; and to characterize phenotypically and genetically these rhizobia. The experiment was carried out in a completely randomized design with five replications. Five to seven months after seedling emergency, according to the legume species, growth and nodulation parameters were determined. Rhizobia identification of strains was carried out by 16S rRNA gene partial sequencing. The survival of seedlings after the transplanting varied from 93 to 98%, in Ultisol (Argissolo in Brazilian classification), collected in primary forest, and fertilized with all nutrients, except nitrogen (ULTfert); and in a clay and sand mixture, in a ratio 3:2 (CONV), respectively. Species with height superior to 30 cm, in general, grew better in substrates with higher fertility: ULTfert and Humic Gley soil (HG). Seven out of the eight species were able to nodulate. The percentage of nodulation per substrate was: SAND, washed sand with mixed inoculum of 100 rhizobia strains plus fertilization (100), HG (80), CONV (100), ULT, A-horizon of red-yellow Ultisol collected in the Ducke Forest Reserve (Manaus) (44), and ULTfert (55%). Bradyrhizobium spp. were isolated from nodules of all species and substrates. Burkolderia fungorum was isolated from Dalbergia inundata. For Dalbergia riedelli and Dalbergia inundata, this is the first report on the identification of symbiotic strains. Scandent legumes present high survival of seedlings in nursery, and develop better in substrates with higher fertility, and generally present symbiosis with [...].(AU)

Um grande número de espécies e indivíduos de leguminosas escandentes estabelecendo simbiose com bactérias fixadoras de nitrogênio ocorrem na floresta Amazônica. Estas simbioses provavelmente desempenham importante papel, contribuindo para incorporação de nitrogênio neste ecossistema. Os objetivos deste trabalho foram avaliar o crescimento de oito espécies de leguminosas escandentes em cinco substratos no viveiro; comparar a nodulação com as estirpes de rizóbio introduzidas ou nativas destes substratos; e caracterizar esses rizóbios fenotípica e geneticamente. O experimento foi conduzido em delineamento inteiramente casualizado com cinco repetições. Cinco a sete meses após a emergência das mudas parâmetros de crescimento e nodulação foram determinados. A identificação das estirpes de rizóbios foi feita pelo sequenciamento parcial do gene 16S rRNA. A sobrevivência das mudas após o transplante variou de 93 a 98%, em um Argissolo (Ultisol de acordo com a classificação da USDA), coletado em floresta primária e adubado com todos os nutrientes, exceto nitrogênio (ULTfert), e na mistura argila e areia, na proporção 3:2 (CONV), respectivamente. Espécies que apresentaram altura superior a 30 cm, no geral, cresceram melhor nos substratos de maior fertilidade: ULTfert e solo Glei Húmico (GH). Das oito espécies estudadas, sete nodularam. A porcentagem de nodulação por substrato foi: AREIA, areia lavada com inóculo misto de 100 estirpes de rizóbio mais adubação (100), GH (80), CONV (100), ULT, horizonte A de Argissolo vermelho-amarelo coletado na Reserva Florestal Ducke (Manaus) (44) e ULTfert (55%). Bradyrhizobium spp. foram isolados de nódulos de todas as espécies e substratos. Burkolderia fungorum foi isolada de Dalbergia inundata. Este é o primeiro relato de identificação de estirpes simbióticas para Dalbergia riedelli e Dalbergia inundata. Leguminosas escandentes apresentaram alta sobrevivência de mudas em casa de vegetação e desenvolveram-se [...](AU)

Nursery growth and rhizobia symbiosis of scandent Leguminosae species native to the Amazon region / Crescimento e simbiose com rizóbios em casa de vegetação de espécies de Leguminosae escandentes nativas da região Amazônica

A great number of species and individuals of scandent legumes establishing symbiosis with nitrogen fixing bacteria occurs in the Amazon Forest. These symbiosis probably play an important role in contributing to nitrogen incorporation in this ecossystem. The objectives of this study were to evaluate the growth of eight species of scandent legumes in five nursery substrates; to compare nodulation with rhizobia strains introduced or native to these substrates; and to characterize phenotypically and genetically these rhizobia. The experiment was carried out in a completely randomized design with five replications. Five to seven months after seedling emergency, according to the legume species, growth and nodulation parameters were determined. Rhizobia identification of strains was carried out by 16S rRNA gene partial sequencing. The survival of seedlings after the transplanting varied from 93 to 98%, in Ultisol (Argissolo in Brazilian classification), collected in primary forest, and fertilized with all nutrients, except nitrogen (ULTfert); and in a clay and sand mixture, in a ratio 3:2 (CONV), respectively. Species with height superior to 30 cm, in general, grew better in substrates with higher fertility: ULTfert and Humic Gley soil (HG). Seven out of the eight species were able to nodulate. The percentage of nodulation per substrate was: SAND, washed sand with mixed inoculum of 100 rhizobia strains plus fertilization (100), HG (80), CONV (100), ULT, A-horizon of red-yellow Ultisol collected in the Ducke Forest Reserve (Manaus) (44), and ULTfert (55%). Bradyrhizobium spp. were isolated from nodules of all species and substrates. Burkolderia fungorum was isolated from Dalbergia inundata. For Dalbergia riedelli and Dalbergia inundata, this is the first report on the identification of symbiotic strains. Scandent legumes present high survival of seedlings in nursery, and develop better in substrates with higher fertility, and generally present symbiosis with [...].

Um grande número de espécies e indivíduos de leguminosas escandentes estabelecendo simbiose com bactérias fixadoras de nitrogênio ocorrem na floresta Amazônica. Estas simbioses provavelmente desempenham importante papel, contribuindo para incorporação de nitrogênio neste ecossistema. Os objetivos deste trabalho foram avaliar o crescimento de oito espécies de leguminosas escandentes em cinco substratos no viveiro; comparar a nodulação com as estirpes de rizóbio introduzidas ou nativas destes substratos; e caracterizar esses rizóbios fenotípica e geneticamente. O experimento foi conduzido em delineamento inteiramente casualizado com cinco repetições. Cinco a sete meses após a emergência das mudas parâmetros de crescimento e nodulação foram determinados. A identificação das estirpes de rizóbios foi feita pelo sequenciamento parcial do gene 16S rRNA. A sobrevivência das mudas após o transplante variou de 93 a 98%, em um Argissolo (Ultisol de acordo com a classificação da USDA), coletado em floresta primária e adubado com todos os nutrientes, exceto nitrogênio (ULTfert), e na mistura argila e areia, na proporção 3:2 (CONV), respectivamente. Espécies que apresentaram altura superior a 30 cm, no geral, cresceram melhor nos substratos de maior fertilidade: ULTfert e solo Glei Húmico (GH). Das oito espécies estudadas, sete nodularam. A porcentagem de nodulação por substrato foi: AREIA, areia lavada com inóculo misto de 100 estirpes de rizóbio mais adubação (100), GH (80), CONV (100), ULT, horizonte A de Argissolo vermelho-amarelo coletado na Reserva Florestal Ducke (Manaus) (44) e ULTfert (55%). Bradyrhizobium spp. foram isolados de nódulos de todas as espécies e substratos. Burkolderia fungorum foi isolada de Dalbergia inundata. Este é o primeiro relato de identificação de estirpes simbióticas para Dalbergia riedelli e Dalbergia inundata. Leguminosas escandentes apresentaram alta sobrevivência de mudas em casa de vegetação e desenvolveram-se [...]

"Influência de aportes de nitrogênio no desenvolvimento inicial de guandu"

O projeto foi desenvolvido na Embrapa Pecuária Sudeste (São Carlos/SP) e dividido em duas etapas. A primeira etapa consistiu em um experimento em campo, onde avaliou-se a influência da inoculação com rizóbios específicos, não específicos e fertilização nitrogenada no desenvolvimento inicial de duas linhagens de guandu. Os tratamentos foram representados por inoculação de sementes com estirpes de Bradyrhizobium SEMIA 6156, recomendada para guandu; estirpes de Bradyrhizobium SEMIA 5079 5080, recomendada para cultura da soja; fertilização nitrogenada na dose de 50 kg.ha-1 N; e tratamento testemunha, aplicados na linhagem g6-95 e cultivar BRS Mandarim. As plantas foram colhidas aos 40 dias após emergência, e avaliadas quanto aos parâmetros de crescimento e acúmulo de N na parte aérea. A segunda etapa foi representada por um experimento em casa de vegetação, com objetivo de avaliar as características relacionadas ao desenvolvimento vegetativo de duas linhagens de guandu sob influência de inoculação com rizóbios específicos, não específicos e doses de adubação nitrogenada combinadas ou não com a inoculação, e seus efeitos sobre a abundância de micro-organismos rizosféricos. Os tratamentos consistiram em duas linhagens de guandu, inoculadas com estirpes SEMIA 6156; estirpes SEMIA 5079 5080; doses de 10 kg.ha-1 N combinadas aos inoculantes; fertilização nitrogenada na dose de 50 kg.ha-1 N; e tratamento testemunha. As plantas foram avaliadas aos 90 dias após emergência, e avaliadas quanto aos parâmetros de crescimento e nodulação. Os resultados demonstraram que não houve diferença entre as linhagens e estirpes avaliadas, indicando as plantas inoculadas com SEMIA 5079 5080, produzem tanto quanto as plantas inoculadas com rizóbio específico, e em condições de campo produzem o equivalente àquelas adubadas com fertilizante nitrogenado. O uso de 10 kg.ha-1 N nas plantas inoculadas possibilitou maior produção de massa seca e melhorou os teores de N da parte aérea. A cultivar BRS Mandarim, apresentou maior nodulação que a linhagem g6-95, no entanto se mostrou menos eficiente na absorção de N, o que pode estar relacionado aos diferentes padrões de exsudação radicular e interações com microbiota do solo rizosférico. No geral, o guandu apresentou baixa especificidade, resultando em índices de eficiência simbiótica elevada mesmo em solos sem inoculação.

The project was developed at Embrapa South-East Cattle Research Center (São Carlos, SP, Brazil) and divided into two experiments. The first experiment consisted of a field experiment, where the influence of inoculation with specific and non-specific rhizobia and nitrogen fertilization on the initial development of two pigeonpea lines was evaluated. The treatments were represented by seed inoculation with Bradyrhizobium strains SEMIA 6156, recommended for pigeon pea; Bradyrhizobium strains SEMIA 5079 5080, recommended for soybean cultivation; nitrogen fertilization at a rate of 50 kg.ha-1 of N; and control treatment, applied to line g6-95 and BRS Mandarim. The plants were harvested 40 days after emergence and evaluated for growth parameters and shoot N accumulation. The second experiment was conducted in a greenhouse, with the objective of evaluating the characteristics related to the vegetative development of two pigeon pea lines under the influence of inoculation with specific and non-specific rhizobia, and nitrogen fertilization rates combined or not with inoculation and its effects on the abundance of rhizospheric microorganisms. The treatments consisted of two pigeon pea lines, inoculated with Bradyrhizobium strains SEMIA 6156; Bradyrhizobium strains SEMIA 5079 5080; nitrogen fertilization at a rate of 10 kg.ha-1 N combined with inoculants; nitrogen fertilization at a rate of 50 kg.ha-1 of N; and control treatment. The plants were evaluated 90 days after emergence and evaluated for growth and nodulation parameters. The results showed that there was no difference between the tested lines and strains evaluated, indicating that plants inoculated with SEMIA 5079 5080 produce as much as plants inoculated with specific rhizobia, and under field conditions they produce the equivalent of those fertilized with nitrogen fertilizer. The use of 10 kg.ha-1 of N in the inoculated plants allowed increase of production of dry mass and better levels of N in the shoot. The BRS Mandarim showed higher nodulation than line g6-95, but was less efficient in N uptake, which may be related to different patterns of root exudation and interactions with the rhizospheric soil organisms. In general, pigeonpea had low specificity, resulting in high rates of symbiotic efficiency even in soils without inoculation.

Produção de biomassa e composição bromatológica da alfafa submetida a coinoculação com bactérias

A alfafa apresenta alto potencial de produção de forragem, qualidade proteica, alta palatabilidade e digestibilidade, capacidade de fixação biológica de nitrogênio no solo e baixa sazonalidade. O objetivo deste trabalho foi avaliar a eficiência de estirpes de Sinorhizobiummeliloti e Azospirillum brasilense, composição bromatológica e na produção de massa seca da parte aérea (MSPA) e massa seca de raízes (MSR) da alfafa cultivada em ArgissoloVermelho Amarelo distrófico. Foi conduzido um ensaio em condições controladas de casa de vegetação. O delineamento experimental foi em blocos casualizados com cinco repetições, com medidas repetidas no tempo. Os tratamentos foram: controle sem N-mineral e sem inoculação; controle com N-mineral e sem inoculação (controle positivo); e os seguintes tratamentos com N-mineral e inoculados, S. melilotiSEMIA 116; S. melilotiSEMIA 116 + coinoculado com A. brasilense (Ab-V5 + Ab-V6); S. melilotiSEMIA 134; S. melilotiSEMIA 134 + coinoculaçãoAb-V5 + Ab-V6; S. melilotiSEMIA 135; S. melilotiSEMIA 135 + coinoculaçãoAb-V5 + Ab-V6. A produção de MSPA por cortes e a MSPA acumulada, para os tratamentos coinoculados com S.melilotiAb-V5 + Ab-V6 produziram 63% e 39% a mais que o controle positivo, respectivamente.Para a MSR acumulada o tratamento coinoculado comS. melilotiSEMIA 134e Ab-V5 + Ab-V6, produziu 64% a mais que o controle positivo. O teor e acúmulo de nitrogênio da parte aérea para o tratamento coinoculado com S. melilotiSEMIA 135e Ab-V5 + Ab-V6, foi 59% e 157% a mais que o controle positivo, respectivamente. Os teores de proteína bruta(PB) e digestibilidade in vitro da matéria seca (DIVMS) para os tratamentos coinoculados com S. melilotiSEMIA 135e Ab-V5 + Ab-V6, apresentou 19% e 13% a mais quando comparado com o controle positivo, respectivamente.A alfafa coinoculada com Ab-V5 + Ab-V6não diferiu das plantas inoculadas somente com as cepas de S. meliloti. No entanto, para a maioria das variáveis foi superior ao tratamento com N e sem inoculação, ou seja, a coinoculação com bactérias promotoras de crescimento vegetal, promoveram resultados positivos no desenvolvimento da alfafa no índice relativo de clorofila, na produção de MSPA, MSR, PB, DIVMS, e no teor acumulado de N da planta e suas raízes

Alfalfa has high forage production potential, protein quality, high palatability and digestibility, capacity for biological nitrogen fixation in the soil and low seasonality. The objective of this work was to evaluate the efficiency of strains of Sinorhizobium meliloti and Azospirillum brasilense, bromatological composition and in the yield of dry matter of the shoot (SDMY) and dry mass of roots (RDM) of the alfalfa cultivated in Dystrophic Red Yellow Argisol. A trial was conducted under controlled greenhouse conditions. The experimental design was in randomized blocks with five repetitions, with repeated measures over time. The treatments were: control without N-mineral and without inoculation; control with N-mineral and without inoculation (positive control); and the following treatments with N-mineral and inoculated, S. meliloti SEMIA 116; S. meliloti SEMIA 116 + co-inoculated with A. brasilense (Ab-V5 + Ab-V6); S. meliloti SEMIA 134; S. meliloti SEMIA 134 + co-inoculated with Ab-V5 + Ab-V6; S. meliloti SEMIA 135; S. meliloti SEMIA 135 + co-inoculated with Ab-V5 + Ab-V6. The production of SDMY by cuts and the accumulated SDMY for the co-inoculated treatments with S. meliloti and Ab-V5 + Ab-V6 produced 63% and 39% more than the positive control, respectively. For accumulated RDM, the co-inoculated treatment with S. meliloti SEMIA 134 plus Ab-V5 + Ab-V6, produced 64% more than the positive control. The nitrogen content and uptake of the aerial part for the co-inoculated treatment with S. meliloti SEMIA 135 and Ab-V5 + Ab-V6, was 59% and 157% more than the positive control, respectively. The crude protein (CP) and in vitro dry matter digestibility (IVDMS) contents for the co-inoculated treatments with S. meliloti SEMIA 135 and Ab-V5 + Ab-V6, showed 19% and 13% more when compared to the control positive, respectively. Co-inoculated alfalfa with A. brasilense Ab-V5 + Ab-V6 did not differ from plants inoculated only with strains of S. meliloti. However, for most variables, it was superior to treatment with N and without inoculation, that is, co-inoculation with plant growth promoting bacteria, promoted positive results in the development of alfalfa in the relative chlorophyll index, in the production of SDMY, RDM, PB, DIVMS, and the N nutrient uptake of the plant and its roots

Symbiotic potential and survival of native rhizobia kept on different carriers

Native rhizobia are ideal for use as commercial legume inoculants. The characteristics of the carrier used to store the inoculants are important for the survival and symbiotic potential of the rhizobia. The objective of this study was to investigate the effects of peat (PEAT), perlite sugarcane bagasse (PSB), carboxymethyl cellulose plus starch (CMCS), and yeast extract mannitol supplemented with mannitol (YEMM) on the survival, nodulation potential and N₂ fixation capacity of the native strains Sinorhizobium mexicanum ITTG R7^T and Rhizobium calliandrae LBP2-1^T and of the reference strain Rhizobium etli CFN42^T. A factorial design (4 × 3) with four repetitions was used to determine the symbiotic potential of the rhizobial strains. The survival of the strains was higher for PEAT (46% for strain LBP2-1^T, 167% for strain CFN42^T and 219% for strain ITTG R7^T) than for the other carriers after 240 days, except for CFN42^T kept on CMCS (225%). All the strains kept on the different carriers effectively nodulated common bean, with the lowest number of nodules found (5 nodules) when CFN42^T was kept on CMCS and with the highest number of nodules found (28 nodules) when ITTG R7^T was kept on PSB. The nitrogenase activity was the highest for ITTG R7^T kept on PEAT (4911 μmol C₂H₄ per fresh weight nodule h⁻¹); however, no activity was found when the strains were kept on YEMM. Thus, the survival and symbiotic potential of the rhizobia depended on the carrier used to store them..(AU)

Plant densities and modulation of symbiotic nitrogen fixation in soybean

Soybean nitrogen (N) demands can be supplied to a large extent via biological nitrogen fixation, but the mechanisms of source/sink regulating photosynthesis/nitrogen fixation in high yielding cultivars and current crop management arrangements need to be investigated. We investigated the modulation of symbiotic nitrogen fixation in soybean [Glycine max (L.) Merrill] at different plant densities. A field trial was performed in southern Brazil with six treatments, including non-inoculated controls without and with N-fertilizer, both at a density of 320,000 plants ha1, and plants inoculated with Bradyrhizobium elkanii at four densities, ranging from 40,000 to 320,000 plants ha1. Differences in nodulation, biomass production, N accumulation and partition were observed at stage R5, but not at stage V4, indicating that quantitative and qualitative factors (such as sunlight infrared/red ratio) assume increasing importance during the later stages of plant growth. Decreases in density in the inoculated treatments stimulated photosynthesis and nitrogen fixation per plant. Similar yields were obtained at the different plant densities, with decreases only at the very low density level of 40,000 plants ha1, which was also the only treatment to show differences in seed protein and oil contents. Results confirm a fine tuning of the mechanisms of source/sink, photosynthesis/nitrogen fixation under lower plant densities. Higher photosynthesis and nitrogen fixation rates are capable of sustaining increased plant growth.