Impacto de herbicidas em isolados de Trichoderma spp.

O uso de microrganismos é uma alternativa para o controle de doenças em plantas. Todavia, é prudente verificar a interação desse com os demais métodos de controle empregados em determinada cultura. Dessa forma, objetivou-se avaliar a fungitoxicidade dos herbicidas sobre o crescimento e desenvolvimento dos isolados de Trichoderma spp. Utilizou-se o delineamento inteiramente casualizado, em esquema fatorial 6 x 6 x 4, com quatro repetições. O fator A correspondeu aos herbicidas pendimethalin, clomazone, carfentrazone-ethyl, oxadiazon, thiobencarb + propanil e byspiribac-sodium; o fator B, às doses dos herbicidas - 0, 25, 50, 75, 100 e 200% da dose recomendada; e o fator C, aos isolados de Trichoderma spp. AJAM 18, CE 66, TRI 01 e TRI 02. O ensaio foi realizado em condições in vitro; avaliaram-se o crescimento micelial radial (CMR) e a esporulação dos isolados após aplicação dos herbicidas. Observaram-se diferenças de sensibilidade dos isolados para o mesmo produto testado. O oxadiazon reduziu o CMR dos isolados AJAM 18 e TRI 01 em 66 e 35%, respectivamente. No entanto, reduziu apenas 16% do CMR do isolado TRI 02 e não alterou o CMR do isolado CE 66 mesmo em 200% da dose recomendada. Verificaram-se diferentes efeitos dos produtos em cada isolado. A mistura comercial de thiobencarb+propanil foi altamente tóxica aos isolados de Trichoderma spp., com reduções em torno de 85% no CMR e no número de esporos. Por outro lado, o byspiribac-sodium pouco afetou os isolados, apresentando reduções inferiores a 10% no CMR e na esporulação. O carfentrazone-ethyl e byspiribac-sodium demonstraram ser compatíveis com os isolados de Trichoderma spp. estudados.

Filtration enrichment method for isolation of auxotrophic mutants of Trichoderma harzianum rifai

A obtenção de marcas genéticas, quer sejam para resistência a drogas, quer para auxotrofia, é uma etapa trabalhosa mas importante em pesquisa genética. Esse trabalho visou a obtenção de mutantes auxotróficos de Trichoderma harzianum utilizando-se a técnica de enriquecimento por filtração. A técnica mostrou-se superior à técnica convencional de isolamento total. Doze mutantes auxotróficos obtidos foram testados quanto a estabilidade, crescimento e resistência ao fungicida benomil. Eles apresentaram taxas de crescimento e esporulação comparáveis à linhagem parental e dois mutantes foram resistentes a benomil em uma concentração de 500µg/ml.

Genetic recombinants in Trichoderma pseudokoningii (Rifai) without typical parasexuality

Heterokaryons were obtained by hyphal anastomosis in Trichoderma pseudokoningii using strains with morphologic and double auxotrophic genetic markers. Stable haploid and unstable hyperhaploid recombinants were detected in monosporic colonies derived from heterokaryons and selected in appropriate media or by an enrichment filtration technique. Diploids were not observed and the recombinant frequencies were higher than those found in fungi with a typical parasexual cycle. The mechanism of recombinant emergence may involve a transient diploid formation followed by chromosomal losses at the hyphal stages, resulting in haploid and hyperhaploid recombinants. The possibility of intrachromosomal rearrangement with the involvement of cytoplasmatic DNA as a heterokaryotic transformation system cannot be excluded.

Atividade antagônica in vitro de isolados de Trichoderma spp. ao fungo Phytophthora citrophthora

Gummosis is among the main fungal diseases of the citrus. It is caused by Phytophthora sp. and usually shows up in the lap of the plant, provoking rottenness and gum exudation, and expands causing the plant death for constrictions in the cambium or phloem which interrupts the descending flow of sap. The objective of this work was to evaluate the antagonistic in vitro activity of Trichoderma spp. to the fungi Phytophthora citrophthora. Phytophthora citrophthora was exposed to five environments of antagonism (without antagonist and with four strains of Trichoderma viride, T. virens, T. harzianu and T stromaticum), The in vitro essay was accomplished through the method of paired cultures. A completely randomized desing was used with five treatments and three replications, and each plot was represented by three petri dishes. The isolates of Trichoderma demonstrated significant effect in the inhibition of the mycelial growth of the fungi Phytophthora citrophthora, and the fungi Trichoderma stromaticum presented larger antagonism to the fungi P. citrophthora while the T harzianum presented antagonism smaller.

Comparison of β-1,3-glucanase production by Botryosphaeria rhodina MAMB-05 and Trichoderma harzianum Rifai and its optimization using a statistical mixture-design

Botryosphaeria rhodina MAMB-05 produced β-1,3-glucanases and botryosphaeran when grown on glucose, while Trichoderma harzianum Rifai only produced the enzyme. A comparison of long-term cultivation (300h) by B. rhodina demonstrated a correlation between the formation of botryosphaeran (48h) and its consumption (after 108h), and de-repression of β-1,3-glucanase synthesis when glucose was depleted from the nutrient medium, whereas for T. harzianum enzyme production commenced during exponential growth. Growth profiles and levels of β-1,3-glucanases produced by both fungi on botryosphaeran also differed, as well as the production of β-1,3-glucanases and β-1,6-glucanases on glucose, lactose, laminarin, botryosphaeran, lasiodiplodan, curdlan, Brewer's yeast powder and lyophilized fungal mycelium, which were dependent upon the carbon source used. A statistical mixture-design used to optimize β-1,3-glucanase production by both fungi evaluated botryosphaeran, glucose and lactose concentrations as variables. For B. rhodina, glucose and lactose promoted enzyme production at the same levels (2.30UmL -1), whereas botryosphaeran added to these substrates exerted a synergic effect favorable for β-glucanase production by T. harzianum (4.25UmL -1). © 2010 Elsevier B.V.

Agaricus blazei as a Substrate for the Production of β-1,3-Glucanase by Trichoderma harzianum Rifai

Extracellular β-1,3-glucanase was produced by Trichoderma harzianum Rifai cultivated in the Agaricus blazei (Agaricus brasiliensis) extract as a substrate in submerged fermentation. A 22-central composite factorial design was developed using the time of culture (x1/day) and Agaricus blazei extract concentration (x2/(g/L)) as variables, and the results were analyzed using response surface methodology (RSM). The results showed that the Agaricus blazei extract concentration was the most important variable in the production of β-1,3-glucanase, and the maximum β-1,3-glucanase activity (0.77 U/mL) was obtained in one day of cultivation. The β-glucan present in the cell wall of Agaricus blazei mushroom proved to be a good substrate for inducing the production of specific β-1,3-glucanase by Trichoderma harzianum Rifai.

Comparing submerged and solid-state fermentation of agro-industrial residues for the production and characterization of lipase by Trichoderma harzianum

Lipase production by Trichoderma harzianum was evaluated in submerged fermentation (SF) and solid-state fermentation (SSF) using a variety of agro-industrial residues. Cultures in SF showed the highest activity (1.4 U/mL) in medium containing 0.5 % (w/v) yeast extract, 1 % (v/v) olive oil and 2.5 C:N ratio. This paper is the first to report lipase production by T. harzianum in SSF. A 1:2 mixture of castor oil cake and sugarcane bagasse supplemented with 1 % (v/w) olive oil showed the best results among the cultures in SSF (4 U/g ds). Lipolytic activity was stable in a slightly acidic to neutral pH, maintaining 50 % activity after 30 min at 50 C. Eighty percent of the activity remained after 1 h in 25 % (v/v) methanol, ethanol, isopropanol or acetone. Activity was observed with vegetable oils (olive, soybean, corn and sunflower) and long-chain triacylglycerols (triolein), confirming the presence of a true lipase. The results of this study are promising because they demonstrate an enzyme with interesting properties for application in catalysis produced by fermentation at low cost. © 2012 Springer-Verlag and the University of Milan.

Small-angle X-ray scattering and structural modeling of full-length: Cellobiohydrolase I from Trichoderma harzianum

Cellobiohydrolases hydrolyze cellulose releasing cellobiose units. They are very important for a number of biotechnological applications, such as, for example, production of cellulosic ethanol and cotton fiber processing. The Trichoderma cellobiohydrolase I (CBH1 or Cel7A) is an industrially important exocellulase. It exhibits a typical two domain architecture, with a small C-terminal cellulose-binding domain and a large N-terminal catalytic core domain, connected by an O-glycosylated linker peptide. The mechanism by which the linker mediates the concerted action of the two domains remains a conundrum. Here, we probe the protein shape and domain organization of the CBH1 of Trichoderma harzianum (ThCel7A) by small angle X-ray scattering (SAXS) and structural modeling. Our SAXS data shows that ThCel7A linker is partially-extended in solution. Structural modeling suggests that this linker conformation is stabilized by inter- and intra-molecular interactions involving the linker peptide and its O-glycosylations. © 2013 Springer Science+Business Media Dordrecht.

Biodiversidade do gênero Trichoderma (Hypocreales - fungi) mediante técnicas moleculares e análise ecofisiográfica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Produção de beta-glucanases por Trichoderma harzianum Rifai para obtenção gluco-oligossacarídeos a partir de botriosferana

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of lettuce bottom rot by isolates of Trichoderma spp

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Production of β-(1,3)-glucanases by Trichoderma harzianum Rifai: Optimization and Application to Produce Gluco-oligosaccharides from Paramylon and Pustulan

β-(1→3)-Glucanases were produced by Trichoderma harzianum Rifai PAMB-86 cultivated on botryosphaeran in a bench-fermenter and optimised by the response surface method. Maximal enzyme titres occurred at 5 days, initial pH 5.5 and aeration of 1.5vvm. β-(1→3)-The β-glucanolytic enzyme complex produced by T. harzianum Rifai PAMB- 86 was fractionated by gel filtration into 2 fractions (F-I, F-II), and employed to produce gluco-oligosaccharides from algal paramylon ((1→3)-β-D-glucan) and lichen pustulan ((1→6)-β-D-glucan). Both enzymes attacked paramylon to the extent of ~15-20% in 30 min releasing glucose and laminaribiose as major end-products, and laminarioligosaccharides of degree of polymerization (DP) ≥3. Only F-I degraded pustulan resulting in ~2% degradation at 30 min, with glucose, gentiobiose and gentio-oligosaccharides of DP ≥4 as major products. The difference in the nature of the hydrolysis products can be explained by the substrate specificities of each enzyme fraction, and the structural differences of the β-D-glucans attacked.

Avaliação de isolados de Trichoderma spp. para controle de Phytophthora nicotianae

Pós-graduação em Microbiologia Agropecuária - FCAV

Unraveling Trichoderma species in attine ant environment: description of three new taxa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de enzimas lignocelulósicas por fermentação em estado sólido por espécimes do gênero Trichoderma para sacarificação do bagaço de cana de açúcar tratado com micro-ondas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)