Resumo O objetivo deste trabalho foi isolar bactérias endofíticas de pupunheira (Bactris gasipaes var. gasipaes) e avaliar os efeitos da inoculação delas em mudas da planta. As bactérias foram isoladas de folhas e raízes das mudas e de meristemas de pupunheira in vitro. Os isolados foram caracterizados fenotipicamente, e, depois, 15 deles, representando grupos fenotípicos distintos, foram selecionados e identificados por meio do sequenciamento parcial do gene 16S rRNA. Em seguida, esses isolados e duas estirpes comerciais de Azospirillum brasilense (Ab-V5 e Ab-V6) foram inoculados em plântulas de pupunheira. Após 76 dias, as mudas foram avaliadas quanto ao desenvolvimento vegetal. Foram identificados os seis seguintes gêneros com base no sequenciamento: Pseudomonas, Enterobacter, Rhizobium, Stenotrophomonas, Klebsiella e Erwinia. Dos 15 isolados inoculados, 9 tiveram efeito positivo sobre a massa de matéria seca de raízes, com destaque para CNPF 77 (Enterobacter sp.), CNPF 100 (Rhizobium sp.), e CNP 179 e CNPF 277 (Stenotrophomonas sp.). Mudas de pupunheira abrigam bactérias endofíticas capazes de aumentar a matéria seca das raízes.

Abstract The objective of this work was to isolate endophytic bacteria from peach palm (Bactris gasipaes var. gasipaes) plants and to evaluate the effects of their inoculation on the plant seedlings. Bacteria were isolated from the leaves and roots of the seedlings and from the meristems of peach palm plants in vitro. The isolates were characterized phenotypically and, then, 15 of them, representing different phenotypic groups, were selected and identified by partial sequencing of the 16S rRNA gene. Afterward, these isolates and two commercial strains of Azospirillum brasilense (Ab-V5 and Ab-V6) were inoculated in the peach palm seedlings. After 76 days, the seedlings were evaluated for plant development. The following six genera were identified based on the sequencing: Pseudomonas, Enterobacter, Rhizobium, Stenotrophomonas, Klebsiella, and Erwinia. Out of the 15 inoculated isolates, 9 had a positive effect on the root dry mass of palm peach, with CNPF 77 (Enterobacter sp.), CNPF 100 (Rhizobium sp.), and CNP 179 and CNPF 277 (Stenotrophomonas sp.) standing out. Peach palm seedlings harbor endophytic bacteria which are able to increase root dry matter.

Resumo: O objetivo deste trabalho foi isolar bactérias endofíticas de cultura de tecidos e de folhas de jabuticabeira (Plinia peruviana) e avaliar seu potencial como bactérias promotoras de crescimento de plantas. As bactérias foram isoladas a partir de segmentos nodais cultivados in vitro e de folhas de árvore sob condições naturais, tendo totalizado 11 e 54 isolados, respectivamente. Os isolados foram caracterizados pela morfologia da colônia. Foram quantificados os compostos indólicos produzidos pelos isolados, na presença ou na ausência de 100 mg L-1 de triptofano. Os maiores produtores destes compostos foram identificados pelo sequenciamento do gene 16S rRNA e foram inoculados em sementes de jabuticaba, tendo-se utilizado Azospirillum brasilense (Ab-V6) como controle positivo. Também avaliou-se a sensibilidade das bactérias a oito antibióticos. Todas as bactérias avaliadas produziram compostos indólicos, especialmente Bacillus sp., com quantidade de 27,41 μg mL-1. A taxa de germinação das sementes inoculadas com Stenotrophomonas sp. foi elevada - 97,34% em comparação à de 74,67% do controle negativo. Bacillus sp. e Stenotrophomonas sp. também aumentaram a velocidade de germinação. O cloranfenicol limitou o crescimento de 82% dos isolados, seguido pelas amoxicilina, gentamicina, levofloxacina e tetraciclina, que limitaram 70%; a eritromicina foi eficaz apenas contra 35%. As bactérias endofíticas isoladas da jabuticaba apresentam características de bactérias promotoras de crescimento de plantas, e Bacillus sp. e Stenotrophomonas sp., provenientes da cultura de tecido, são capazes de aprimorar a germinação de sementes de jabuticaba.

Abstract: The objective of this work was to isolate endophytic bacteria from tissue culture and leaves of jaboticaba (Plinia peruviana) and to evaluate their potential as plant growth-promoting bacteria. The bacteria were isolated from nodal segments grown in vitro and from leaves from a tree under natural conditions, totaling 11 and 54 isolates, respectively. The isolates were characterized by colony morphology. The indolic compounds produced by the isolates, in the presence or absence of 100 mg L-1 tryptophan, were quantified. The greatest producers of these compounds were identified by sequencing the 16S rRNA gene and were inoculated on jaboticaba seeds, using Azospirillum brasilense (Ab-V6) as a positive control. The sensitivity of bacteria to eight antibiotics was also evaluated. All assessed bacteria produced indolic compounds, especially Bacillus sp., with a content of 27.41 μg mL-1. The germination rate of the seeds inoculated with Stenotrophomonas sp. was high - 97.34% compared with that of 74.67% of the negative control. Bacillus sp. and Stenotrophomonas sp. also sped up germination. Chloramphenicol limited the growth of 82% of the isolates, followed by amoxicillin, gentamicin, levofloxacin, and tetracycline, which limited 70%; erythromycin was only effective against 35%. The endophytic bacteria isolated from jaboticaba show characteristics of plant growth-promoting bacteria, and Bacillus sp. and Stenotrophomonas sp., obtained from tissue culture, are capable of enhancing jaboticaba seed germination.

RESUMO A castanha-do-brasil é um produto florestal não madeireiro muito importante na região amazônica. A propagação desta árvore ainda representa um desafio, devido ao lento e irregular processo de germinação das sementes. Neste contexto, bactérias promotoras do crescimento vegetal podem facilitar o processo de propagação. O objetivo deste estudo foi isolar e caracterizar bactérias endofíticas em raízes de castanha-do-Brasil em floresta de terra firme e em áreas cultivadas no norte do Brasil, e identificar alguns mecanismos de promoção do crescimento vegetal executados por essas bactérias. No total, 90 isolados bacterianos foram obtidos de raízes de castanha-do-Brasil em monocultura, agrofloresta e floresta nativa, usando diferentes meios de cultivo semi-sólidos. Os isolados foram caracterizados pelo sequenciamento do gene 16S rRNA. As características de promoção do crescimento vegetal foram avaliadas através da presença do gene nifH, solubilização de fosfato de alumínio e produção de compostos indólicos. Os isolados foram afiliados a 18 gêneros, pertencentes a cinco diferentes classes (α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Bacilli e Actinobacteria). O gênero Bacillus foi predominante, principalmente nas áreas de floresta e monocultura. Quatorze isolados apresentaram o gene nifH. A maioria dos isolados foi capaz de solubilizar fosfato de alumínio e sintetizar compostos indólicos. Os resultados indicam uma elevada diversidade de bactérias endofíticas presente em raízes de castanha-do-Brasil, principalmente em área de agrofloresta, que pode estar relacionado aos atributos do solo. Entre os 90 isolados, 22 apresentaram os melhores resultados relacionados às características de promoção do crescimento vegetal, e são bons candidatos para testes em produção de mudas de castanha-do-Brasil.

ABSTRACT Brazil nut is a very important nontimber forest product in the Amazon region. Propagation of this tree still represents a challenge due to slow and uneven seed germination. In this context, plant growth-promoting bacteria can facilitate the process of propagation. The aims of this study were to isolate and characterize endophytic bacteria from the roots of Brazil nut trees in native terra firme forest and cultivation areas in northern Brazil, and to identify mechanisms by which bacteria act in plant growth promotion. Overall, 90 bacterial isolates were obtained from the roots of Brazil nut trees in monoculture, agroforestry and native forest areas by using different semisolid media. The isolates were characterized by sequencing the 16S rRNA gene. Plant growth-promoting characteristics were evaluated by the presence of the nifH gene, aluminum phosphate solubilization and the production of indole compounds. The isolates were affiliated with 18 genera belonging to 5 different classes (α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Bacilli and Actinobacteria). The genus Bacillus was predominant in the forest and monoculture areas. Fourteen isolates presented the nifH gene. Most of the bacteria were able to solubilize aluminum phosphate and synthetize indole compounds. The results indicated high diversity of endophytic bacteria present among the roots of Brazil nut trees, mainly in the agroforestry area, which could be related to soil attributes. Among the 90 isolates, the 22 that presented the best results regarding plant growth promotion traits were good candidates for testing in seedling production of Brazil nut trees.

Resumo O presente estudo teve como objetivos determinar a melhor condição de sombreamento e o tipo de substrato para a produção de mudas e estabelecimento da simbiose espontânea por rizóbios em taxi-branco (Tachigali vulgaris). O experimento foi instalado no delineamento de blocos casualizados, com quatro repetições e os tratamentos arranjados em esquema de parcela subdividida, no qual a parcela consistiu de dois níveis de sombreamento (50% e 75%) e a subparcela de quatro tipos de substrato: solo; solo + areia (1:1); solo + areia + serragem (2:1:1); e solo + vermiculita (1:1). Cada parcela experimental foi constituída por cinco mudas. As mudas foram avaliadas quanto a um conjunto de variáveis morfológicas e fisiológicas, da parte aérea e da raiz. No final do experimento foi também realizado o isolamento e a caracterização fenotípica de bactérias oriundas de nódulos coletados nas mudas. A caracterização foi realizada pela avaliação morfológica das colônias. O sombreamento de 50% é o indicado na produção de mudas de taxi-branco, enquanto que, em relação ao substrato, aquele composto por solo + areia é o que proporciona mudas de melhor qualidade. A espécie apresentou simbiose com grande diversidade de rizóbios, apesar de o número de isolados não ter sido influenciado pelo tipo de substrato e nível de sombreamento. A maioria dos isolados obtidos apresentam tempo de crescimento muito rápido e pH ácido do meio de cultura.

Abstract This study had as objectives to determine the best shading condition and the type of substrate to produce seedlings and the establishment of a spontaneous symbiosis by rhizobia in taxi-branco (Tachigali vulgaris). A randomized block design with four replications and treatments arranged in a split-plot scheme was installed. The plot consisted of two shading levels (50% and 75%) and subplots of four types of substrates: soil; soil + sand (1:1); soil + sand + sawdust (2:1:1); and soil + vermiculite (1:1). Each experimental plot consisted of five seedlings. The seedlings were evaluated for a set of shoot and root morphological and physiological traits. At the end of the experiment, the isolation and the phenotypic characterization of bacteria from nodules collected in the seedlings were also carried out. The characterization was performed by the morphological evaluation of the colonies. The shading of 50% is indicated for seedling productions, while in relation to the substrate, that one composed by soil + sand is what produces seedlings of better quality. Although the number of isolates was not influenced by the substrate type and the shading level, taxi-branco seedlings establish symbiosis with a great diversity of rhizobia. Most of the isolates have very fast growth rate and acid pH of the culture medium.

ABSTRACT The use of dark septate fungi (DSE) to promote plant growth can be beneficial to agriculture, and these organisms are important allies in the search for sustainable agriculture practices. This study investigates the contribution of dark septate fungi to the absorption of nutrients by rice plants and their ensuing growth. Four dark septate fungi isolates that were identified by Internal transcribed spacer phylogeny were inoculated in rice seeds (Cv. Piauí). The resulting root colonization was estimated and the kinetic parameters Vmax and Km were calculated from the nitrate contents of the nutrient solution. The macronutrient levels in the shoots, and the NO3--N, NH4+-N, free amino-N and soluble sugars in the roots, sheathes and leaves were measured. The rice roots were significantly colonized by all of the fungi, but in particular, isolate A103 increased the fresh and dry biomass of the shoots and the number of tillers per plant, amino-N, and soluble sugars as well as the N, P, K, Mg and S contents in comparison with the control treatment. When inoculated with isolates A103 and A101, the plants presented lower Km values, indicating affinity increases for NO3--N absorption. Therefore, the A103 Pleosporales fungus presented the highest potential for the promotion of rice plant growth, increasing the tillering and nutrients uptake, especially N (due to an enhanced affinity for N uptake) and P.

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

ABSTRACT Inga edulis Mart. is a leguminous tree adapted to acidic and low-fertility soils that establishes symbioses with nitrogen (N)-fixing bacteria. The identification of effective bacteria in biological N fixation may bolster the use of I. edulis in degraded or modified areas and agroforestry systems. Therefore, the aims of this study were evaluation of the symbiotic effectiveness of eight strains of the Bradyrhizobium genus native to Roraima in Inga edulis plants, and in vitro evaluation of the ability of the eight strains of Bradyrhizobium to develop plant growth-promoting characteristics. Determination of symbiotic effectiveness was carried out via three experiments: the first in a greenhouse in pots with a sterile substrate; the second in a greenhouse in pots containing non-sterile soil; and the third in a nursery in bags with a non-sterile substrate. Twelve treatments were evaluated: inoculation with eight strains of Bradyrhizobium ingae (ERR 490, ERR 492, ERR 493, ERR 494T, ERR 496, ERR 497, ERR 498, and ERR 569); inoculation with two strains indicated for Inga marginata, BR 6609 and BR 6610 (positive controls); no inoculation but with mineral N; and neither inoculation nor mineral N. All of the experiments were conducted in a completely randomized design with four replicates. The first experiment was conducted for 60 days, and the other experiments were conducted for 100 days. For all of the experiments, the number of nodules, nodule dry matter, root dry matter, shoot dry matter, number of leaflets, plant height, stem diameter, total N in the shoots, root/shoot dry matter ratio, Dickson’s quality index, relative effectiveness, and the Pearson correlation between the variables under study were evaluated. The strains were also evaluated by their ability to solubilize calcium and aluminum phosphates and to produce indolic compounds. The results showed that B. ingae strains were effective in biological N fixation, especially the ERR 493, ERR 498, and ERR 569 strains. These strains increased the production of shoot dry matter and total N and exhibited relative effectiveness higher than 100 % in all of the experiments. The B. ingae strains were also able to solubilize calcium and aluminum phosphates, despite their synthesis of indolic compounds. Thus, the strains of B. ingae can be used for inoculation in the production of I. edulis seedlings.

RESUMO Plantas de milho podem estabelecer associações benéficas com bactérias promotoras do crescimento vegetal. No entanto, poucos estudos de caracterização e inoculação com essas bactérias foram realizados na região Amazônica. O objetivo deste estudo foi caracterizar bactérias endofíticas isoladas de milho na região Amazônica e avaliar sua capacidade de promover o crescimento. Cinquenta e cinco isolados bacterianos foram obtidos de plantas de milho cultivadas em diferentes tipos de ecossistemas, cerrado (savana) e área de floresta. Estes foram caracterizados pela presença do gene nifH, capacidade de sintetizar ácido indol-3-acético (AIA), solubilizar fosfato de cálcio (CaHPO4) e sequenciamento parcial do gene 16S rRNA. Vinte e quatro bactérias apresentaram o gene nifH. Destas, sete são oriundas de plantas cultivadas em uma área de cerrado e 17 em área de floresta. Quatorze bactérias demonstraram a capacidade de solubilizar AIA e quatro de solubilizar fosfato de cálcio. Entre estas bactérias, foram encontrados os seguintes gêneros: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia e Bacillus. Oito isolados com capacidade de promoção do crescimento vegetal foram selecionados para experimento em casa de vegetação com a inoculação em dois diferentes genótipos de milho (um híbrido e uma variedade) em vasos com solo. Quanto à promoção de crescimento vegetal, a inoculação promoveu o desenvolvimento de plantas de milho, mas não foi encontrada interação significativa entre a cultivar de milho e a inoculação bacteriana. Na região Amazônica há uma diversidade de bactérias endofíticas e estas possuem potencial para promover o desenvolvimento de plantas de milho.

ABSTRACT Maize plants can establish beneficial associations with plant growth-promoting bacteria. However, few studies have been conducted on the characterization and inoculation of these bacteria in the Amazon region. This study aimed to characterize endophytic bacteria isolated from maize in the Amazon region and to assess their capacity to promote plant growth. Fifty-five bacterial isolates were obtained from maize grown in two types of ecosystems, i.e., a cerrado (savanna) and a forest area. The isolates were characterized by the presence of the nifH gene, their ability to synthesize indole-3-acetic acid (IAA) and solubilize calcium phosphate (CaHPO4), and 16S rRNA partial gene sequencing. Twenty-four bacteria contained the nifH gene, of which seven were isolated from maize plants cultivated in a cerrado area and seventeen from a forest area. Fourteen samples showed the capacity to synthesize IAA and only four solubilized calcium phosphate. The following genera were found among these isolates: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia and Bacillus. In addition, eight isolates with plant growth-promoting capacity were selected for a glasshouse experiment involving the inoculation of two maize genotypes (a hybrid and a variety) grown in pots containing soil. Inoculation promoted the development of the maize plants but no significant interaction between maize cultivar and bacterial inoculation was found. A high diversity of endophytic bacteria is present in the Amazon region and these bacteria have potential to promote the development of maize plants.

O objetivo deste trabalho foi isolar e caracterizar rizóbios de nódulos de Centrolobium paraense e avaliar sua eficiência simbiótica. Amostras de solo, coletadas de quatro locais do Cerrado de Roraima, foram utilizadas para o cultivo de C. paraense, para obtenção dos nódulos. Os isolados (178) foram obtidos dos 334 nódulos coletados após o cultivo em meio 79. Vinte e cinco isolados, pertencentes a seis grupos morfológicos, foram autenticados com uso de Vigna unguiculata e caracterizados pelo 16S rRNA. Os isolados identificados como Bradyrhizobium foram caracterizados depois pelo sequenciamento do gene rpoB. Um experimento em casa de vegetação foi realizado com C. paraense, para testar os 18 isolados autenticados. Aproximadamente 90% dos isolados mostraram crescimento lento em meio 79. A análise do 16S rRNA mostrou que 14 dos isolados autenticados pertencem ao gênero Bradyrhizobium, e o rpoB indicou que eles constituem grupos diferentes em comparação às espécies já descritas. Somente quatro dos 11 isolados com crescimento rápido nodularam V. unguiculata, dois dos quais pertencentes a Rhizobium e dois a Pleomorphomonas, que não foi relatado anteriormente como gênero nodulífero. Os isolados de Bradyrhizobium ERR 326, ERR 399 e ERR 435 apresentaram a maior eficiência em C. paraense e mostraram contribuição similar ao tratamento nitrogenado. Centrolobium paraense é capaz de nodular com diferentes espécies de rizóbios, algumas das quais ainda não foram descritas.

The objective of this work was to isolate and characterize rhizobia from nodules of Centrolobium paraense and to evaluate their symbiotic efficiency. Soil samples collected from four sites of the Roraima Cerrado, Brazil, were used to cultivate C. paraense in order to obtain nodules. Isolates (178) were obtained from 334 nodules after cultivation on medium 79. Twenty-five isolates belonging to six morphological groups were authenticated using Vigna unguiculata and they were characterized by 16S rRNA. Isolates identified as Bradyrhizobium were further characterized using rpoB gene sequencing. A greenhouse experiment was carried out with C. paraense to test the 18 authenticated isolates. Approximately 90% of the isolates grew slowly in medium 79. The 16S rRNA analysis showed that 14 authenticated isolates belong to the genus Bradyrhizobium, and rpoB indicated they constitute different groups compared to previously described species. Only four of the 11 fast-growing isolates nodulated V. unguiculata, two of which belong to Rhizobium, and two to Pleomorphomonas, which was not previously reported as a nodulating genus. The Bradyrhizobium isolates ERR 326, ERR 399, and ERR 435 had the highest symbiotic efficiency on C. paraense and showed a contribution similar to the nitrogen treatment. Centrolobium paraense is able to nodulate with different rhizobium species, some of which have not yet been described.

A fixação biológica de nitrogênio que ocorre em leguminosas é realizada por um grupo de bactérias conhecidas como rizóbios. A sobrevivência destas bactérias no solo é influenciada por diversos fatores como a temperatura, umidade e fertilidade do solo. O objetivo deste trabalho foi avaliar a dinâmica da população de rizóbios em solo após o cultivo de soja, durante o período de estiagem no cerrado de Roraima. Foram amostradas três áreas: i) cerrado nativo como referência; ii) área cultivada uma vez com soja inoculada com rizóbio; iii) e área cultivada duas vezes com soja inoculada com rizóbio em anos consecutivos. O solo foi coletado na profundidade de 0-10 cm em cinco períodos a partir do inicio da estiagem no mês de setembro de 2006 coincidindo com a época de colheita da soja e prolongando-se até março de 2007 (0, 45, 90, 135 e 180 dias). A população de rizóbios no solo foi avaliada pela técnica do número mais provável (NMP) utilizando plantas de soja e de feijão-caupi como espécies isca. Foi observado que na área nativa praticamente não existiam bactérias nodulantes de soja, mas havia uma população capaz de nodular o feijão-caupi de até algumas centenas de rizóbios por gramas de solo. O cultivo da soja utilizando sementes inoculadas elevou a população de rizóbios no solo que foi constatada por ambas às espécies de plantas isca. Nas áreas cultivadas, constatou-se uma intensa redução da população de rizóbios no solo, em especial logo após a colheita da soja, continuando o decréscimo até o último período de avaliação. Conclui-se que o cultivo da soja inoculada com rizóbio eleva a densidade de rizóbios em solo do cerrado, mas durante a estiagem ocorre uma drástica redução dessa população, que pode chegar a mais de 99% considerando o início e final do período.

The biological nitrogen fixation in legumes is performed by a group of bacteria known as rhizobia. The survival of these bacteria in soils is affected by several factors, such as temperature, drought and soil fertility. This study was performed to evaluate the dynamics of rhizobia in the soil after soybean cultivation and during a dry season in the cerrado of Roraima. Three areas were sampled: i) native cerrado as reference; ii) an area previously cultivated with soybean for one season; and iii) another one cultivated for two seasons also with soybean. The soil was sampled at a depth of 0-10 cm in five times (0, 45, 90, 135 and 180 days) during the dry season (September 2006 to March 2007). The rhizobial density in the soil was evaluated by the most probable number method with infection of soybean and cowpea plants. It was observed very low number of soybean nodulating bacteria in the reference area, but a high density, of up to several hundred rhizobia capable to nodulate cowpea was measured in this same area. Cropping of soybean with inoculated seeds increased rhizobial density evaluated by both trapping hosts. In cropped areas, an intense reduction of rhizobium density was observed just after soybean harvest, and this reduction continued until the end of the period of evaluation. It was concluded that soybean cultivation increases the density of rhizobial in the cerrado soil; however, this density is drastically reduced, during the dry season, by 99% at the end of the dry period.

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.

Non-symbiotic diazotrophic bacteria are amongst the most important functional groups of soil-dwelling microorganisms. These bacteria contribute to plant growth predominantly through biological N2 fixation. Here, we evaluated the density and diversity of non-symbiotic diazotrophic bacteria in soils taken from diverse land use systems (LUS) in Amazonia using nitrogen-free media. A total of 30 soil samples were collected from the following LUS: pristine forest, young secondary forest, old secondary forest, agroforestry, agriculture and pasture. Bacterial density was evaluated by the most probable number (MPN) method utilizing N-free semi-solid media with varied compositions (JNFb, NFb, LGI and Fam). Individual isolates were characterized by colony and cellular morphology as well as total protein profiles and nitrogenase activity. Isolate genotypes were determined by partial 16S rDNA sequences. No typical diazotrophic growth in the JNFb medium was observed. Bacterial densities in the NFb medium were higher in the agriculture and agroforestry soil samples. In LGI and Fam media, bacterial densities were highest in the pasture soil samples. Overall, 22 isolates with high phenotypic diversity were obtained. Eleven isolates exhibited nitrogenase activity. Sequences of 16S rDNA genes of 14 out of 19 isolates had similarities below 100 % with known nitrogen-fixing species. Isolates were identified as belonging to the Burkholderia, Enterobacter, Serratia, Klebsiella, and Bacillus genera. A higher number of isolates from pasture soil samples were isolated, with the majority of these belonging to the Burkholderia and Bacillus genera. Among the isolates, unknown sequences were obtained, possibly indicating new species. Taken together, these data demonstrate that Fam, NFb, and LGI semi-solid media allowed the growth of diazotrophic bacteria belonging to different phylogenetic lines.

Apesar de o zinco (Zn) ser micronutriente fundamental para o crescimento e metabolismo das plantas, quando presente em níveis tóxicos no ambiente pode afetar o desenvolvimento vegetal. Entre os vários efeitos benéficos do silício (Si), cita-se sua influência na diminuição ou eliminação dos efeitos adversos de metais pesados no meio. O objetivo deste trabalho foi avaliar o efeito do Si na amenização da toxidez de Zn sobre o crescimento e nutrição mineral de plantas de Eucalyptus urophylla. As plantas foram cultivadas em vasos contendo 3 L de solução nutritiva de Clark, em esquema fatorial 6 x 2, sendo seis concentrações de Zn (0, 2, 50, 150, 300 e 450 µmol L-1 como ZnSO4 7H2O) e duas de Si (0 e 1,78 mmol L-1 de Si como silicato de potássio). Após oito semanas, avaliaram-se alguns parâmetros morfológicos das plantas, produção de matéria seca, teores e utilização de nutrientes. O aumento das concentrações de Zn na solução nutritiva proporcionou maior fitotoxicidade nas raízes em relação à parte aérea. A adição do Si amenizou o efeito negativo do excesso de Zn sobre o crescimento, no entanto pouco influenciou os teores dos nutrientes avaliados nos tecidos, embora tenha proporcionado utilização mais eficiente de P, Ca, Mg e S pelas plantas de Eucalyptus urophylla.

Zn is an essential micronutrient for the growth and metabolism of plants, but when present in toxic levels in the environment, it can affect the the development of plants. Among the several beneficial effects of silicon (Si), it presents an effect in the decrease or elimination of the adverse effects of heavy metals in the environment. The objective of this work was to evaluate the effect of Si in the alleviation of Zn toxicity on the growth and mineral nutrition of Eucalyptus urophylla seedlings cultivated in nutrient solution. Seedlings of Eucalyptus urophylla were grown in pots containing 3L of Clark nutrient solution, in a 6 x 2 factorial design, using six Zn concentrations (0, 2, 50, 150, 300 and 450 µmol L-1 as ZnSO4 7H2O) and two of Si (0 and 1.78 mmol L-1 of Si as potassium silicate). After eight weeks, plant morphologic traits, dry matter production and content and nutrient utilization were evaluated. The increase of Zn concentrations in the nutrient solution provided higher toxicity in the roots than in the shoots. The addition of the Si alleviated the negative effect of the excess Zn on plant growth, but presented little effect on the nutrient content in these tissues. The silicon addition in the nutrient solution provided a more efficient use of P, Ca, Mg and S for the Eucalyptus urophylla plants.

A espécie de leguminosa Sesbania virgata estabelece uma simbiose especifica e eficiente com Azorhizobium doebereinerae. Estudos prévios indicam que a ocorrência de A. doebereinerae esta relacionada à presença de S. virgata. Avaliou-se a ocorrência de A. doebereinerae e de outras Bactérias Fixadoras de Nitrogênio Nodulíferas em Leguminosas (BFNNL) em amostras de solos coletadas próximo e a 10 metros do caule de cinco plantas de S. virgata em áreas de pastagem. As características simbióticas de isolados das populações dessas BFNNL foram também estudadas. Para captura de A. doebereinerae e de outras BFNNL, essas amostras de solos foram inoculadas em S. virgata e nas leguminosas promíscuas Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris e Vigna unguiculata. Todas as amostras de solos apresentaram BFNNL capazes de nodular, pelo menos, uma espécie. M. atropurpureum foi a espécie mais promíscua capturando um grande número de tipos culturais de BFNNL das suspensões de solo. As outras espécies foram menos promíscuas na seguinte ordem: V. unguiculata, P. vulgaris, e L. leucocephala. Os isolados das espécies hospedeiras consideradas promíscuas foram agrupados em sete tipos culturais. Um desses grupos, isolado de todas as espécies promiscuas, apresentou crescimento rápido e reação alcalina em meio de cultura (como Azorhizobium) e foi identificado como Cupriavidus. Este é o primeiro relato da simbiose de Cupriavidus com espécies de Papilionoideae. A simbiose de BFNNL com as demais espécies teve eficiência variável, mas foi sempre menor que a da adubação nitrogenada e da estirpe recomendada como inoculante. S. virgata formou simbiose somente com A. doebereinerae, a qual foi mais freqüente nas amostras de solos coletadas próximo ao caule da planta, corroborando alto estímulo deste microsimbionte por sua espécie hospedeira. Uma alta diversidade de BFNNL ocorre saprofiticamente próximo ao sistema radicular de S. virgata.

The legume species Sesbania virgata establishes a specific and efficient symbiosis with Azorhizobium doebereinerae. Previous studies have shown that A. doebereinerae occurrence correlates to the presence of S. virgata. This work aimed to evaluate the occurrence of A. doebereinerae and of other nitrogen-fixing Leguminosae-nodulating bacteria (NFLNB) in soil samples collected adjacent to and 10 m away from the stems of five S. virgata plants in pasture areas. Symbiotic characteristics of isolates from these NFLNB populations were also studied. S. virgata and the four promiscuous legume species Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris and Vigna unguiculata were inoculated with soil samples to trap A. doebereinerae and other NFLNB. NFLNB capable of inducing nodulation in at least one of these legumes were found in all samples. M. atropurpureum was the most promiscuous species, as it trapped the highest number of NFLNB cultural types from soil suspensions. The other species were less promiscuous in the following order: V. unguiculata, P. vulgaris, and L. leucocephala. Isolates of the promiscuous legumes were classified into seven cultural groups. One of these groups, isolated from all promiscuous species, showed fast-growth alkali-reaction in culture medium (like Azorhizobium); it was identified as Cupriavidus. This is the first report of symbiosis of Cupriavidus with Papilionoideae species. The symbiotic efficiency of promiscuous hosts with NFLNB varied, but it was always less than that of controls with mineral nitrogen or an inoculant strain. S. virgata was efficiently nodulated only by A. doebereinerae, which occurred mainly in samples collected close to the plant stem, corroborating a high stimulus by its host species. A high diversity of NFLNB occurs as saprophytes close to the S. virgata root system.