RESUMO

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.

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Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.(AU)

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The aim of this research was to evaluate the efficiency of aqueous alkali-treated Brachiaria straw for the cultivation of appropriate species of oyster mushroom. The substrate used in the cultivation of various Pleurotus spp. was soaked for 20min by using two different procedures: (i) 0.5-2.0% Ca(OH)2 in 100L water, and (ii) 50-250L water. As a result, 1% Ca(OH)2 dissolved in 100L water and 3.5kg of Brachiaria straw presented the best production. The most suitable species for the application of the present method were P. pulmonarius and P. sapidus. The success of this technique is directly related to the concentration of Ca(OH)2 and water, the species, and the origin and quality of raw material used as the substrate in the production of oyster mushroom.

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Abstract The aim of this research was to evaluate the efficiency of aqueous alkali-treated Brachiaria straw for the cultivation of appropriate species of oyster mushroom. The substrate used in the cultivation of various Pleurotus spp. was soaked for 20 min by using two different procedures: (i) 0.5-2.0% Ca(OH)2 in 100 L water, and (ii) 50-250 L water. As a result, 1% Ca(OH)2 dissolved in 100 L water and 3.5 kg of Brachiaria straw presented the best production. The most suitable species for the application of the present method were P. pulmonarius and P. sapidus. The success of this technique is directly related to the concentration of Ca(OH)2 and water, the species, and the origin and quality of raw material used as the substrate in the production of oyster mushroom.

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Brachiaria brizantha is considered one of the preferred fodders among farmers for having high forage yield and large production of root mass. The association of beneficial bacteria with these grasses can be very valuable in the recovery of the pasture areas with nutritional deficiency. With the aim of studying this possibility, we carried out the sampling of soil and roots of B. brizantha in three areas (Nova Odessa-SP, São Carlos-SP and Campo Verde-MT, Brazil). Seventy-two bacterial strains were isolated and used in tests to evaluate their biotechnological potential. Almost all isolates presented at least one positive feature. Sixty-eight isolates produced analogues of indole-3-acetic acid, ten showed nitrogenase activity when subjected to the method of increasing the concentration of total nitrogen (total N) in the culture medium and sixty-five isolates showed nitrogenase activity when subjected to acetylene reduction technique. The partial sequencing of 16S rRNA of these isolates allowed the identification of seven main groups, with the prevalence of those affiliated to the genus Stenotrophomonas (69 %). At the end, this work elected the strains C4 (Pseudomonadaceae) and C7 (Rhodospirillaceae) as promising organisms for the development of inoculants due to their higher nitrogenase activity.

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Os sistemas silvipastoris têm despertado interesse por permitir amplas possibilidades de combinar plantas no espaço e no tempo com múltiplos atributos. O Brasil possui a maior variedade de espécies florestais nativas do planeta e condições climatológicas favoráveis à produção de forragens tropicais. Dessa forma, objetivou-se avaliar parâmetros produtivos e morfofisiológicos de Brachiaria decumbens em relação aos renques de árvores de um sistema silvipastoril implantado com espécies arbóreas nativas e sua relações com variáveis microclimáticas (transmissividade da radiação fotossinteticamente ativa e armazenamento de água no solo). Para isso foi conduzido um experimento no campo experimental da Embrapa Pecuária Sudeste, em São Carlos SP, entre novembro de 2009 e abril de 2011. O delineamento experimental foi aleatorizado em blocos completos. O efeito da distância das árvores sobre a massa de material morto, densidade, massa de forragem, massa de folha e de colmos, índice de área foliar, área foliar específica, relação folha:colmo, transmissividade e armazenamento de água foi avaliada ao longo das estações do ano. A interação distância x estação influenciou todas as variáveis analisadas da forragem e a transmissividade da radiação fotossinteticamente ativa. As variações dos níveis de armazenamento de água no solo entre as distâncias foram semelhantes, no entanto observou-se uma variação em relação ao volume em algumas épocas, principalmente entre os perfis.

The silvopastoral systems have attracted interest because it allows a lot of possibilities of combining plants in space and time with multiple attributes. Brazil has the largest variety of native species on the planet and favorable climatological conditions for the production of tropical forages. This work aimed to evaluate the productive and morphophysiological Brachiaria decumbens in relation to the rows of trees in a silvopastoral system deployed with native tree species and their relationships with microclimatic variables (transmissivity of photosynthetically active radiation and soil water storage). For this purpose an experiment was conducted at the experimental field of Embrapa Southeast Cattle in Sao Carlos - SP, between November 2009 and April 2011. The experimental design was randomized complete block. The effect of distance from trees on the mass of dead material, density, herbage mass, leaf and stem, leaf area index, specific leaf area, leaf: stem ratio, transmissivity and storage of water was evaluated along the seasons. The interaction distance x season influenced all variables of forage and transmissivity of photosynthetically active radiation. Changes in levels of water storage in soil between the distances are similar, however there was a variation in the volume at some times, especially between the profiles. The average transmission of photosynthetic active radiation was 38, 59, 81 and 89% for the distances 0.0, 2.0, 4.75 and 8.5 m, for tutors trees respectively. Among the larger distances only difference was in the autumn when transmissivity of 4.75 m was 12% lower compared to the distance of 8.5 m.

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Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants.

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AIM: To evaluate the effects of soil physical and chemical factors (pH, conductivity, humidity, available phosphorus and organic matter) and environmental factors (temperature, relative air humidity, altitude and atmospheric pressure) on arbuscular mycorrhizal fungi (AMF)-Brachiaria decumbens grass relationship. Furthermore to establish patterns of microbiological responses that allow to differentiate the study sites in two relief types. METHODS AND RESULTS: Mycorrhizal characteristics (spore density, external hyphae and root colonizations by hyphae, vesicles and arbuscules), physical and chemical factors in soil and environmental factors were measured. CONCLUSIONS: The effect of physical, chemical and environmental factors on microbiological variables was related to the type of relief 'valley and hilly terrain'; the AMF behaviour was affected only over narrower ranges of evaluated variables. Similarly, the colonization of B. decumbens roots by AMF hyphae, vesicles and the mycorrhizal spore density follow different patterns according to the relief type. SIGNIFICANCE AND IMPACT OF THE STUDY: The type of relief is one of the factors to be taken into consideration to evaluate the AMF inoculum and root colonization of these pastures, because of the influence of slope - as physical property of soil - on AMF.

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Brachiaria, predominantly an African genus, contains species, such as B. brizantha, an apomictic C4 grass, that are commercially important forage grasses in tropical America, where they now cover about 55 million hectares. From B. brizantha accession CIAT 6780, we isolated an endophytic fungus that may be economically significant. The fungus was identified as Acremonium implicatum (J. Gilman & E.V. Abott). 18S rDNA and ITS rDNA sequences were used to characterize isolates of the endophyte, and showed that they belonged to the Acremonium genus, being close to A. strictum and A. kiliense. Using the random amplified polymorphic DNA (RAPD) technique, involving arbitrary primers of 10 bases, we showed that the isolates were highly similar to each other. Antiserum produced from a monoconidial culture of A. implicatum isolated from B. brizantha 6780, differentiated the isolates consistently in line with the DNA data. When we compared endophyte-free with endophyte-infected B. brizantha CIAT 6780 plants, both artificially inoculated with the pathogenic Drechslera fungus, we found that the endophyte-infected plants had fewer and smaller lesions than did the endophyte-free plants. Sporulation of Drechslera sp. on artificially inoculated leaf sheath tissues was also much less on tissue infected with the endophyte.

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