Abstract Vegetative desiccation tolerance (VDT) is a biological process that allows resurrection plants to complete desiccation-rehydration cycles and survive without permanent damage. Physiological and biochemical changes related to the VDT were evaluated in the lycophyte Selaginella convoluta from the Caatinga biome, a seasonally dry tropical forest of northeastern Brazil. As representative of the oldest division of extant vascular plants, the investigation of VDT mechanisms in this species serves as an opportunity to provide information for research communities interested in functional traits related to water-limited conditions. The experiments were performed with plants in their natural habitat, in dry and rainy seasons, and in pots under shade house conditions. Relative water content, chlorophyll a fluorescence, gas exchange, activity of antioxidant enzymes, concentration of chlorophylls, proteins, total amino acids, proline, and soluble carbohydrates in leaf tissues were evaluated throughout the process of complete dehydration and subsequent re-watering of intact plants. The overall results reveal the capacity of S. convoluta to tolerate extreme dehydration and rapidly recover upon rehydration. This species minimizes potential damages during the desiccation-rehydration cycle due to the presence of inductive mechanisms (photochemical preventing, sucrose accumulation) and constitutive mechanisms (antioxidant activity of catalase and ascorbate peroxidase), which share similarities with the VDT of both angiosperms and bryophytes.

Resumo Tolerancia à dessecação vegetativa (TDV) é um processo biológico que permite que plantas revivescentes sobrevivam a ciclos completos de dessecação-reidratação sem danos permanentes. Alterações fisiológicas e bioquímicas relacionadas à TDV foram avaliadas na licófita Selaginella convoluta do bioma Caatinga, em uma floresta tropical sazonalmente seca do nordeste do Brasil. Como representante da mais antiga divisão de plantas vasculares não extintas, a investigação de mecanismos de TDV nesta espécie serve como uma oportunidade para fornecer informações para comunidades de pesquisa interessadas em características funcionais relacionadas às condições limitantes de água. Os experimentos foram conduzidos com plantas em seu habitat natural, nas estações seca e chuvosa, e em potes sob casa de vegetação sombreada. Conteúdo relativo de água, fluorescência da clorofila a, trocas gasosas, atividade de enzimas antioxidantes, concentração de clorofilas, proteínas, aminoácidos totais, prolina e carboidratos solúveis de tecidos foliares foram avaliados ao longo dos processos de completa dessecação e subsequente reidratação de plantas intactas. Os resultados revelam a capacidade de S. convoluta para tolerar desidratação extrema e rapidamente recuperar após a reidratação. Esta espécie minimiza danos potenciais durante o ciclo de dessecação-reidratação devido à presença de mecanismos protetivos dos tipos indutivos (inibição fotoquímica, acúmulo de sacarose) e constitutivos (atividade antioxidante de catalase e ascorbato peroxidase), mostrando similaridades com a TDV de angiospermas e briófitas.

RESUMO O tratamento de sementes com fungicidas é uma prática importante para o controle de fitopatógenos na cultura do amendoim. Todavia, esses produtos podem prejudicar rizobactérias (Bradyrhizobium) e inibir processos como a fixação biológica de nitrogênio. Objetivou-se verificar os efeitos do tratamento de sementes de amendoim cv. BR1 inoculadas com estirpes de Bradyrhizobium spp. com fungicidas. O experimento foi conduzido em casa-de-vegetação, utilizando-se duas combinações de fungicidas [C1: carboxin (200 g L-1) + tiram (200 g L-1); C2: pyraclostrobin (25 g L-1) + thiophanate methyl (225 g L-1) + fipronil (250 g L-1)] e uma testemunha sem fungicida; dois inoculantes à base de Bradyrhizobium spp. (SEMIA 6144 e ESA 123) e uma testemunha com fonte química nitrogenada (nitrato de amônio). O delineamento experimental foi inteiramente casualizado, em esquema fatorial 3 (2 com fungicida e 1 sem fungicida) x 4 (2 inoculações à base de rizóbios, 1 fonte mineral de N e 1 sem N) x 2 (1 ou 2 aplicações de inoculantes), com 5 repetições. Foram avaliados a massa seca da raiz e da parte aérea, altura de plantas, nodulação e acúmulo de nitrogênio nas folhas. A aplicação de ambas as combinações reduziu o número de nódulos nas raízes das plantas, principalmente para C1. O crescimento vegetativo e o nitrogênio nas folhas foram afetados por C1, enquanto, para C2, houve incrementos superiores aos da testemunha sem fungicidas. ESA 123 foi mais tolerante aos efeitos dos fungicidas na nodulação. Em relação ao número de inoculações, observou-se incremento na taxa de N com duas inoculações. C2, apesar de afetar a nodulação, foi menos tóxica aos inoculantes SEMIA 6144 e ESA 123.

ABSTRACT Seed treatment with fungicides is an important practice for the control of phytopathogens in peanut crops. However, these products can harm rhizobacteria (Bradyrhizobium) and inhibit processes such as biological nitrogen fixation. This study aimed to verify the effects of the treatment of peanut seeds cv. BR1 inoculated with Bradyrhizobium spp. with fungicides. The experiment was conducted in a greenhouse, using two combinations of fungicides [C1: carboxin (200 g L-1) + tiram (200 g L-1); C2: pyraclostrobin (25 g L-1) + thiophanate methyl (225 g L-1) + fipronil (250 g L-1)] and one control without fungicide; two inoculants based on Bradyrhizobium spp. (SEMIA 6144 and ESA 123) and one control with a nitrogen chemical source (ammonium nitrate). The experimental design was completely randomized, in a 3 (2 with fungicide and 1 without fungicide) x 4 (2 inoculations based on rhizobia, 1 N mineral source and 1 without N) x 2 (1 or 2 inoculants applications) factorial scheme, with 5 replications. The root and shoot dry mass, plant height, nodulation and leaf nitrogen accumulation were evaluated. The application of both combinations reduced the number of nodules on plant roots, mainly for C1. The vegetative growth and nitrogen in the leaves were affected by C1, whereas, for C2, there were increments higher than for the control without fungicides. ESA 123 was more tolerant to the effects of fungicides on nodulation. Regarding the number of inoculations, there was an increase in the nitrogen rate with two inoculations. C2, despite affecting the nodulation, was less toxic to the inoculants SEMIA 6144 and ESA 123.

Abstract 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.

Leguminosas forrageiras são importantes recursos na região semi-árida, devido a sua capacidade de adaptação a solos de baixa fertilidade e fixação de nitrogênio quando associados a bactérias diazotróficas. Aqui, aplicamos uma abordagem polifásica com o objetivo de caracterizar bactérias isoladas de nódulos obtidos de Clitoria ternatea e Stylosanthes capitata cultivadas em solos do semiárido brasileiro. O isolamento das bactérias produziu uma coleção de 126 isolados: 45 isolados de C. ternatea e 81 de S. capitata. Os testes de nodulação resultaram em apenas dez isolados que nodularam seu hospedeiro original: seis de C. ternatea e quatro de S. capitata. Esses dez isolados de nódulos de leguminosas tiveram suas características fenotípicas e genotípicos avaliadas. Todos os isolados cresceram em frutose, glicose, glutamato de sódio, maltose, xilose e sacarose. Testes metabólicos mostraram uma relação entre a tolerância ao sal e altas temperaturas; onde o isolado que tolerou a maior concentração de sal também tolerou a temperatura mais alta. Três isolados apresentaram atividade amilolítica e quatro foram capazes de produzir carboximetil celulolítico. A estreptomicina foi o antibiótico mais limitante para o crescimento bacteriano, e o ácido nalidíxico foi o menos limitante. Sete dos dez isolados eram produtores de IAA. O sequenciamento parcial do gene 16S rRNA permitiu identificar isolados como membros dos gêneros Bacillus (1), Bradyrhizobium (4), Leifsonia (3), Microvirga (1) e Rhizobium (1). Esses dados revelam diversidade fenotípicas e genotípicas de bactérias que habitam nódulos das forrageiras C. ternatea e S. capitata, e representam uma importante fonte microbiana para a prospecção de novos produtos biotecnológicos para promover melhor desenvolvimento das leguminosas forrageiras na região semiárida.

Forage legumes are important resources in semiarid regions due to their abilities to adapt to soils with low fertility and fix nitrogen when associated with diazotrophic bacteria. Here, we applied a polyphasic approach to characterize a set of legume nodule isolates obtained from Clitoria ternatea and Stylosanthes capitata cultivated in the soils of a semiarid region of Brazil. A total of 126 bacterial isolates were obtained: 45 isolates from C. ternatea and 81 isolates from S. capitata. Nodulation tests revealed only ten isolates that nodulated their original host: six isolates from C. ternatea and four isolates from S. capitate. These ten legume nodule isolates were phenotypically and genotypically characterized. All isolates grew in fructose, glucose, sodium glutamate, maltose, xylose, and sucrose. Metabolic tests showed a relationship between tolerance to salt and high temperatures, where isolates that tolerated the highest salt concentration also tolerated the highest temperature. Three isolates showed amylolytic activity, and four isolates showed carboxymethyl cellulolytic activity. Streptomycin was the most limiting and nalidixic acid was the least limiting antibiotic to bacterial growth. Seven out of ten isolates were indol-acetic acid producers. Additionally, 16S rRNA gene partial sequencing enabled the identification of members of the genera Bacillus (1), Bradyrhizobium (4), Leifsonia (3), Microvirga (1), and Rhizobium (1). These data reveal phenotypically and genotypically diverse bacteria inhabiting the nodules of the forage legumes C. ternatea and S. capitata represent an important microbial source to protect new biotechnological products and improve forage legumes in semiarid regions.

ABSTRACT Cowpea is a very important crop to Brazilian Semi-Arid mainly small family-based farmers. Rhizobia inoculation is a practice, easy to use, and cheap technology that increases cowpea productivity. The aim of this study was to evaluate the efficiency of two new rhizobia isolates in greenhouse and field as well as classify them taxonomically. To bacterial identification the 16S rRNA gene of ESA 17 and ESA 18 isolates were sequenced. The greenhouse test was conducted with pots containing 3 L of soil and the bacterial isolates evaluated were ESA 17, ESA 18, BR 3267 or BR 3262 strains. A field experiment was implemented on a Vertisol in Juazeiro, Bahia State, to evaluate the cowpea growth and productivity. In this experiment, the peat-based inoculants with ESA 17, ESA 18, BR 3267 or UFLA 3-84 were used in 2 cowpea cultivars. Both bacteria were identified as Bradyrhizobium, but related to different species. ESA 17 was related to B. japonicum and ESA 18 was closer to B. pachyrhizi. At greenhouse, both isolates increased cowpea nitrogen content in the shoots due to the presence of very efficient nodules. In the field, the isolate ESA 18 inoculated at BRS Pujante cultivar induced higher production than observed for the absolute control, and for BR 17 Gurguéia cultivar, the ESA 17 and BR 3267 stood out both by inducing high production and grain protein content. The results indicate that both isolates can be evaluated in network experiments aiming at official recommendation for new bacteria to cowpea inoculant in Brazil.

ABSTRACT Legume-rhizobia symbiosis is a cross-kingdom association that results in large amounts of nitrogen incorporated in food webs. For the Brazilian semi-arid region, data on genetic variability and symbiotic efficiency of Papilionoidae rhizobial communities are very scarce. The aim of this study was to evaluate the genetic variability and the symbiotic efficiency of eight rhizobial isolates obtained from “mulungu” (Erythrina velutina Willd.) nodules. For 16S rRNA gene sequencing, the genomic DNA was extracted using a commercial kit, amplified with universal primers, and subjected to sequencing reactions. For the isolate ESA 71, PCR amplifications for nodC and nodA genes were attempted. Rhizobial efficiency was assessed by two greenhouse experiments. The first assay was carried out under gnotobiotic conditions, with sterile sand as a substrate; the second experiment was conducted in a non-sterile soil. For both experiments, the inoculation treatments consisted of a single inoculation of each isolate, in addition to a treatment with Bradyrhizobium elkanii BR 5609 as a reference strain. Furthermore, two non-inoculated control treatments, supplied and not supplied with mineral N, were also evaluated. Bacterial identification indicated that both α and β-rhizobia could be found in “mulungu” root nodules. Three isolates where classified within the Rhizobium genus, four bacteria belonged to Bradyrhizobium and one isolate clustered with Burkholderia. Positive amplification of an intragenic fragment of the nodA gene using a primer set to β-rhizobia could be found for ESA 71 (Burkholderia). All bacterial isolates were effective in colonizing “mulungu” roots. In the first experiment, all inoculated treatments and N fertilization increased the N concentration in “mulungu” shoot tissues. For total N in the shoots, the isolates ESA 70, ESA 72, and ESA 75 stood out. In the non-sterile substrate experiment, the isolates ESA 70, ESA 71, ESA 72, and ESA 75, together with the reference strains, induced increases in the shoot N concentration and total accumulation compared to the absolute control. The results indicate that “mulungu” is able to establish associations with efficient α and β-rhizobia in Brazilian semi-arid soils.

O objetivo desse estudo foi avaliar as características fenotípicas dos isolados de nódulos de mulungu (Erythrina velutina Willd.) e determinar seu fingerprinting molecular por meio de Box-PCR. Todas as bactérias foram avaliadas pelas seguintes características fenotípicas: tempo de crescimento, alteração de pH, cor da colónia e produção de muco. As bactérias isoladas hábeis em renodular o hospedeiro original também foram avaliadas em relação à tolerância ao aumento da salinidade e da temperatura de incubação, capacidade de crescimento utilizando diferentes fontes de carbono, resistência intrínseca aos antibióticos e biossíntese de auxina “in vitro”. O fingerprint molecular foi estabelecido usando a técnica Box-PCR e os isolados foram agrupados pelos perfis obtidos. Dos 22 isolados obtidos, oito foram hábeis em renodular o hospedeiro original. Entre eles, alguns foram tolerantes a 1% de NaCl e a 39° C e todos metabolizaram as fontes maltose, frutose, glicose, sacarose e arabinose, resistiram à rifampicina e produziram auxina. As bactérias isoladas mostraram baixa similaridade genética entre elas e as estirpes de referência, o que indica a grande variabilidade genética dos isolados. Os resultados deste trabalho constituem o primeiro relato sobre bactérias capazes de nodular esta espécie endémica. O estudo mais aprofundado destas bactérias poderá revelar a existência de isolados tolerantes a stresses ambientais e adequados a serem utilizados como futuros inoculantes de mulungu.

This study aims to evaluate the phenotypical characteristics of bacterial isolates from mulungu (Erythrina velutina Willd.) nodules and determinate their Box-PCR fingerprinting. All bacteria were evaluated by the following phenotypic characteristics: growth rate, pH change, colony color and mucus production. The bacterial isolates able to re-nodulate the original host were also evaluated regarding its tolerance to increased salinity and different incubation temperatures, ability to growth using different carbon sources, intrinsic antibiotic resistance and “in vitro” auxin biosynthesis. The molecular fingerprints were set up using the Box-PCR technique and the isolates were clustered by its profiles. Among the 22 bacterial isolates obtained, eight were able to re-nodulate the original host. Among the nodule inducing isolates, some were tolerant to 1% of NaCl and 39° C and all of them metabolized the maltose, fructose, glucose, sucrose and arabinose, were resistant to rifampicin and produced auxin. The bacteria showed low genetic similarity among them and reference strains, which indicates the great genetic variability of the isolates. The results of this work are the first reports about the bacterial isolates able to nodulate this endemic specie. A more deep study of these bacteria may reveal the existence of isolates tolerant to environmental stresses and suitable as a future mulungu inoculant.

As espécies vegetais que ocorrem naturalmente na Caatinga brasileira apresentam diversas adaptações às condições adversas que podem ser exploradas para aumentar a produção vegetal. Entre essas estratégias adaptativas para o enfrentamento da baixa disponibilidade hídrica, a tolerância à dessecação pode ser destacada. Até o momento, a associação entre essas espécies e os microrganismos benéficos do solo não é bem entendida. Os objetivos deste trabalho foram caracterizar bactérias diazotróficas de Tripogon spicatus e avaliar a capacidade de promoção do crescimento delas em plantas de arroz. Dezesseis isolados bacterianos foram estudados quanto a seu posicionamento taxonômico por meio do sequenciamento parcial do gene 16S rRNA, potencial diazotrófico, produção de ácido indol acético (AIA) e solubilização de fosfato de cálcio in vitro, metabolismo de nove diferentes fontes de C em meio semissólido, tolerância a diferentes concentrações de NaCl a pHs e resistência intrínseca a nove antibióticos. A habilidade em promover o crescimento vegetal foi avaliada utilizando o arroz (Oryza sativa) como planta-modelo. Dentre os 16 isolados avaliados, oito foram classificados como pertencentes à família Enterobacteriaceae, relacionados aos gêneros Enterobacter e Pantoea. Seis outros isolados foram relacionados a Bacillus e outros dois a Rhizobium e Stenotrophomonas. A avaliação da incorporação do N ao meio semissólido demonstrou o potencial diazotrófico de todos os isolados. Duas bactérias produziram mais AIA do que a estirpe BR 11175T de Herbaspirillum seropedicae. Isolados bacterianos foram capazes de formar película em meio semissólido suplementado com concentrações de NaCl de até 1.27 mol L-1. A resistência intrínseca a antibióticos e o metabolismo de diferentes fontes de C indicaram um perfil fisiológico muito variável. Sete isolados foram capazes de aumentar o crescimento das plantas de arroz, mais eficientemente do que a estirpe de referência Ab-V5 de Azospirillum brasilense. Os resultados indicaram o potencial de T. spicatus como uma espécie nativa fonte de bactérias promotoras de crescimento vegetal.

Plant species that naturally occur in the Brazilian Caatinga(xeric shrubland) adapt in several ways to these harsh conditions, and that can be exploited to increase crop production. Among the strategic adaptations to confront low water availability, desiccation tolerance stands out. Up to now, the association of those species with beneficial soil microorganisms is not well understood. The aim of this study was to characterize Tripogon spicatusdiazotrophic bacterial isolates from the Caatingabiome and evaluate their ability to promote plant growth in rice. Sixteen bacterial isolates were studied in regard to their taxonomic position by partial sequencing of the 16S rRNA gene, putative diazotrophic capacity, in vitro indole-acetic acid (IAA) production and calcium phosphate solubilization, metabolism of nine different C sources in semi-solid media, tolerance to different concentrations of NaCl to pHs and intrinsic resistance to nine antibiotics. Finally, the ability of the bacterial isolates to promote plant growth was evaluated using rice (Oryza sativa) as a model plant. Among the 16 isolates evaluated, eight of them were classified as Enterobacteriaceae members, related to Enterobacter andPantoeagenera. Six other bacteria were related toBacillus, and the remaining two were related toRhizobiumand Stenotrophomonas.The evaluation of total N incorporation into the semi-solid medium indicated that all the bacteria studied have putative diazotrophic capacity. Two bacteria were able to produce more IAA than that observed for the strain BR 11175Tof Herbaspirillum seropedicae.Bacterial isolates were also able to form a microaerophilic pellicle in a semi-solid medium supplemented with different NaCl concentrations up to 1.27 mol L-1. Intrinsic resistance to antibiotics and the metabolism of different C sources indicated a great variation in physiological profile. Seven isolates were able to promote rice growth, and two bacteria were more efficient than the reference strainAzospirillum brasilense, Ab-V5. The results indicate the potential of T. spicatus as native plant source of plant growth promoting bacteria.

O objetivo deste trabalho foi avaliar a contribuição de estirpes de rizóbio, eficientes na fixação de nitrogênio, sobre a produção de grãos de novas cultivares de feijão-caupi, indicadas para cultivo no Semiárido brasileiro, no Submédio do Vale do Rio São Francisco. Dois experimentos foram implantados nos perímetros irrigados de Mandacaru (Juazeiro, BA) e Bebedouro (Petrolina, PE). Os tratamentos consistiram da inoculação isolada de cinco estirpes de rizóbio - BR 3267, BR 3262, INPA 03-11B, UFLA 03-84 (Bradyrhizobiumsp.) e BR 3299T (Microvirga vignae) -, além de um tratamento com nitrogênio e de um controle sem inoculação ou aplicação de N. As seguintes cultivares de feijão-caupi foram avaliadas: BRS Pujante, BRS Tapaihum, BRS Carijó e BRS Acauã. Utilizou-se o delineamento experimental de blocos ao acaso, com quatro repetições. As plantas inoculadas apresentaram produtividade de grãos similar à observada em plantas adubadas com 80 kg ha-1 de N. As cultivares BRS Tapaihum e BRS Pujante destacaram-se quanto à produtividade e ao teor de proteínas nos grãos, quando inoculadas, o que mostra seu potencial para cultivo na região do Submédio do Vale do São Francisco.

The objective of this work was to evaluate the contribution of efficient nitrogen-fixing rhizobial strains to grain yield of new cowpea cultivars, indicated for cultivation in the Brazilian Semiarid region, in the sub-medium of the São Francisco River Valley. Two experiments were set up at the irrigated perimeters of Mandacaru (Juazeiro, state of Bahia) and Bebedouro (Petrolina, state of Pernambuco). The treatments consisted of single inoculation of five rhizobial strains - BR 3267, BR 3262, INPA 03-11B, UFLA 03-84 (Bradyrhizobiumsp.), and BR 3299T(Microvirga vignae) -, besides a treatment with nitrogen and a control without inoculation or N application. The following cowpea cultivars were evaluated: BRS Pujante, BRS Tapaihum, BRS Carijó, and BRS Acauã. A randomized complete block design, with four replicates, was used. Inoculated plants showed similar grain yield to the one observed with plants fertilized with 80 kg ha-1 N. The cultivars BRS Tapaihum and BRS Pujante stood out in grain yield and protein contents when inoculated, showing their potential for cultivation in the sub-medium of the São Francisco River Valley.

La asociación de gramíneas silvestres con bacterias diazotróficas en los biomas brasileños es poco conocida. El aislamiento y caracterización de las bacterias asociadas con gramíneas silvestres puede contribuir a entender la ecología de las diazotróficas y bacterias con aplicaciones biotecnológicas. En este estudio, caracterizamos aislamientos bacterianos de diazotróficas de Oryza glumaepatula recolectadas en Cerrado y zonas boscosas de la Amazonía en el estado de Roraima, Brasil. Plantas sanas de O. glumepatula fueron recolectadas en cinco zonas boscosas y siete en Cerrado. Las plantas de Cerrado fueron recolectadas en septiembre 2008, mientras que las del bosque en Junio 2008 y Abril 2009. Las plantas y el suelo adherido a las raíces se transfirieron a macetas y se cultivaron durante 35 días en condiciones de invernadero. Durante la cosecha, los brotes y las raíces se trituraron por separado en una solución salina, la suspensión se diluyó en serie y se inocularon en placas Petri que contenían medio Dyg. Todas las colonias de bacterias se purificaron en el mismo medio. Se evaluó la capacidad diazotrófica de cada bacteria en condiciones microaerofílicas en medio semisólido BMGM. Además, los aislamientos bacterianos que formaron películas se evaluaron también mediante amplificación por PCR para el gen nifH. La diversidad de bacterias nifH+ se analizó por Huella Genética utilizando la Reacción en Cadena de la Polimerasa. Para las cepas seleccionadas, la capacidad de promover el crecimiento de O. sativa como modelo de planta también se evaluó. Se obtuvo un total de 992 cepas bacterianas. Cincuenta y un bacterias fueron capaces de formar películas en el medio semisólido y 38 amplificaron positivamente el fragmento 360bp del gen nifH. De los 38 aislamientos de nifH+, 24 fueron obtenidos de los brotes, mientras que 14 se originaron a partir de las raíces. Los perfiles de PCR-Box mostraron que los aislamientos bacterianos obtenidos en este estudio presentaron una baja similitud con las cepas de referencia pertenecientes a Herbaspirillum, Azospirillum y el género Burkholderia. La capacidad promotora del crecimiento fue confirmada por al menos cinco aislamientos. Para esta bacteria, la raíz y brote mostraron resultados de crecimiento mayores en comparación con los observados en las plantas inoculadas con las cepas de referencia. Estos resultados indican que O. glumaepatula es colonizada por una muy diversa comunidad diazotrófica en la Amazonia brasileña. Se están llevando a cabo otras investigaciones para esclarecer la taxonomía de estas cepas y sus mecanismos para promover el crecimiento.

The association of wild grasses with diazotrophic bacteria in Brazilian biomes is poorly understood. The isolation and characterization of bacteria associated with wild grasses can contribute to understand the diazotrophic ecology as well as to identify bacteria with biotechnological applications. In this study, we isolated and characterized diazotrophic bacterial isolates from Oryza glumaepatula collected in Cerrado and Forest areas of the Amazon in Roraima State, Brazil. Healthy O. glumepatula plants were collected at five sampling sites at Forest and seven at Cerrado, respectively. The plants were collected at the Cerrado areas in September 2008 while the Forest plants were collected in June/2008 and April/2009. The plants and the soil adhering to the roots were transferred to pots and grown for 35 days in greenhouse conditions. During the harvest, the shoots and the roots were crushed separately in a saline solution; the suspension was diluted serially and inoculated in Petri dishes containing Dyg’s medium. All distinct bacterial colonies were purified in the same medium. The diazotrophic capacity of each bacterium in microaerophilic conditions was assessed in semisolid BMGM medium. In addition, the pellicles forming bacterial isolates were also evaluated by PCR amplification for nifH gene. The diversity of nifH+ bacteria was analyzed by Box-PCR fingerprinting. For selected strains, the growth promoting capacity of O. sativa as a model plant was also evaluated. A total of 992 bacterial isolates were obtained. Fifty- one bacteria were able to form pellicles in the semisolid medium and 38 also positively amplified the 360bp nifH gene fragment. Among the 38 nifH+ isolates, 24 were obtained from the shoots, while 14 originated from the roots. The Box-PCR profiles showed that the bacterial isolates obtained in this study presented a low similarity with the reference strains belonging to the Herbaspirillum, Azospirillum and Burkholderia genus. The growth- promoting ability was confirmed for at least five isolates. For these bacteria, the root and shoot growing results showed higher increases when compared to those observed in plants inoculated with the evaluated reference strains. These results indicate that O. glumaepatula is colonized by a high diverse diazotrophic community in the Brazilian Amazon. Further investigations are now being carried out to determine the taxonomic positions of these isolates and their growth promoting mechanisms.

O objetivo deste trabalho foi avaliar a compatibilidade do tratamento de sementes com fungicidas e a inoculação com estirpes de Bradyrhizobium em feijão-caupi. Em laboratório, avaliou-se a sobrevivência de células nas sementes da cultivar BRS Guariba, tratadas ou não com fungicidas (fludioxonil, carbendazim, carbendazim + thiram e carboxin + thiram) e inoculadas ou não com Bradyrhizobium (estirpes UFLA3-84, BR 3267, INPA3-11B e BR 3262). Em casa de vegetação, conduziu-se experimento em vasos de Leonard, com os mesmos tratamentos. Foram avaliados: massa de matéria seca da parte aérea, além de número e massa de nódulos 25 dias após a emergência das plantas. No campo, dois experimentos foram conduzidos, tendo-se utilizado a estirpe BR 3262, com aplicação de fungicidas nas sementes: um em área de primeiro cultivo e outro em área cultivada anteriormente com culturas anuais. Avaliaram-se, aos 35 dias, o número de nódulos, a massa de nódulos secos e a massa de matéria seca da parte aérea, e, na colheita, a produtividade de grãos. Os fungicidas não tiveram efeito significativo sobre a sobrevivência de Bradyrhizobium, a nodulação das plantas e o rendimento de grãos, que, em média, foi superior a 1.200 kg ha-1. O tratamento de sementes de feijão-caupi com fungicidas é compatível com a inoculação das estirpes avaliadas.

The objective of this work was to evaluate the compatibility of seed treatment with fungicides and Bradyrhizobium strains inoculation for cowpea. Cell survival on seeds of cultivar BRS Guariba treated or not with fungicides (fludioxonil, carbendazim, carbendazim + thiram, and carboxin + thiram) and inoculated or not with Bradyrhizobium (strains UFLA3-84, BR 3267, INPA3-11B, and BR 3262) were evaluated in laboratory. Another experiment was carried out in greenhouse, in Leonard jars, using the same treatments. The following were evaluated: shoot dry matter weight, besides number and dry matter weight of nodules 25 days after plant emergence. Two experiments were performed in the field with the strain BR 3262, with fungicides on the seeds: one in a first cultivation area and the other in an area previously cultivated with annual crops. Nodule number, nodule dry weight, and shoot dry matter weight were evaluated at 35 days, and grain yield at harvest. Fungicides did not have significant effect on Bradyrhizobium survival, plant nodulation, and grain yield, which was, in average, higher than 1,200 kg ha-1. Cowpea seed treatment with fungicides is compatible with the inoculation of the evaluated strains.

This study evaluated 26 pigeonpea rhizobial isolates according to their cultural characteristics, intrinsic antibiotic resistance, salt and temperature tolerance, carbon source utilization and amylolytic activity. The cultural characterization showed that the majority of them presented the ability to acidify the YMA. Among the 27 isolates evaluated, 25 were able to grow when incubated at 42° C and 11 showed tolerance to 3% (w/v) of NaCl in YMA medium. The patterns of carbon sources utilization was very diverse among the isolates. It was observed the capacity of three strains to metabolize all the carbon sources evaluated and a total of 42% of the bacterial isolates was able to grow in the culture medium supplemented with at least, six carbon sources. The carbon sources mannitol (control) and sucrose were metabilized by all isolates evaluated. The profile of intrinsic resistance to antibiotics showed that the isolates were mostly resistant to streptomycin and ampicillin, but susceptible to kanamycin and chloranphenicol. High amylolytic activity of, at least, four isolates was also demonstrated, especially for isolated 47.3b, which showed the highest enzymatic index. These results indicate the metabolic versatility of the pigeonpea rhizobia, and indicates the isolate 47.3b to further studies regarding the amylase production and characterization.

As espécies de mucuna são muito utilizadas como adubos verdes, e poucas informações estão disponíveis a respeito dos rizóbios nativos capazes de nodulá-las. O objetivo deste trabalho foi avaliar a diversidade e a capacidade simbiótica de isolados bacterianos de nódulos de mucuna-cinza (Mucuna pruriens (L.) DC.) e mucuna-anã (Mucuna deeringiana (Bort.) Merr.). As bactérias foram isoladas de nódulos de mucunas cinza e anã cultivadas em vasos com solos de um sistema de produção agroecológica. Foram isoladas 160 bactérias, sendo 80 de mucuna-anã e 80 de mucuna-cinza, que foram autenticadas e selecionadas para avaliação da capacidade simbiótica. A diversidade dos isolados foi avaliada por meio das características culturais em meio de cultura YMA e da técnica de análise de restrição do produto de PCR do gene 16S rDNA. A inoculação de cinco isolados em mucuna-cinza e dois em mucuna-anã apresentou elevada biomassa da parte aérea. A maioria dos isolados apresentou crescimento rápido e acidificou o meio de cultura. A análise de restrição demonstrou que as bactérias isoladas apresentam baixa similaridade com estirpes de referência, sugerindo a existência de isolados pertencentes a novos grupos, capazes de nodular as mucunas anã e cinza.

Mucuna species are widely used as green manure, and little information is available about rhizobia that can nodulate them. The objective of this study was to evaluate the diversity and ability of bacteria isolated from symbiotic nodules of velvet bean (Mucuna pruriens (L.) DC.) and Mucuna deeringiana (Bort.) Merr. From the nodules of either mucuna species, grown in pots with soils from an agroecological system, 80 bacteria were isolated. The selected bacteria were classified and used to assess the symbiotic capacity. The diversity assessment of the isolates was based on cultural characteristics on YMA medium and the technique of restriction analysis of PCR products of 16S rDNA. The inoculation of Mucuna pruriens with five and Mucuna deeringiana with two isolates increased the aboveground biomass. Most isolates grew rapidly and acidified the culture medium. Restriction analysis showed low similarity of the reference strains with the bacterial isolates, suggesting the existence of isolates belonging to new groups that are able to nodulate the two mucuna species.

O objetivo deste trabalho foi avaliar a eficiência agronômica da formulação polimérica IPC 2.2 como veículo de inoculação para feijão-caupi. Foi avaliada a sobrevivência de células aos 180 dias de armazenamento. Em experimento de campo, comparou-se a formulação polimérica com a turfosa e a líquida e com adubação nitrogenada de 50 e 80 kg ha-¹ de N e um controle absoluto. A formulação IPC 2.2 manteve concentração de células superior a 109 g-1 após 180 dias e, em campo, proporcionou produtividade de grãos superior à do controle absoluto e igual à dos tratamentos nitrogenados e veículos turfoso e líquido.

The objective of this work was to evaluate the agronomic efficiency of the polymer formulation IPC 2.2 as a carrier for inoculation in cowpea. Cell survival at 180 days storage was evaluated. The polymer formulation was compared with peat and liquid carriers and with nitrogen rates of 50 and 80 kg ha-1 N and absolute control, in a field experiment. The formulation IPC 2.2 kept a cell concentration higher than 10(9) g-1, after 180 days and, in the field experiment, it provided a yield higher than that of the treatment absolute and equal to those of the nitrogen treatments and to the peat and liquid carriers.

O objetivo deste trabalho foi avaliar a ocorrência de fungos endofíticos "dark septate" (DSEF) em Oryza glumaepatula, na Amazônia, e sua capacidade de colonização in vitro. Foram coletadas plantas de O. glumaepatula em área de cerrado e de mata em Roraima. As raízes foram tratadas para a observação de hifas melanizadas septadas e de microescleródios. Os fungos foram isolados em meio ágar malte. Os DSEF foram observados em plantas coletadas em ambos os ambientes, com maior colonização nas coletadas da mata. Um isolado foi capaz de colonizar o hospedeiro original e também plantas de Oryza sativa, exibindo as estruturas características de DSEF em plantas de arroz saudáveis.

The objective of this work was to assess the occurrence of dark septate endophyte fungi (DSEF) in Oryza glumaepatula in Amazonia, and its in vitro colonization capacity. Oryza glumaepatula plants were collected in forest and cerrado areas of the Roraima state, Brazil. The roots were prepared to observe septate melanized hyphas and microesclerotia. The isolation of fungi was done in agar malte medium. Dark septate endophyte fungi were observed in plants from both environments, with higher colonization on those collected in the forest. One isolate was able to colonize the original host and also Oryza sativa plants, exhibiting typical DSEF structures in rice healthy plants.