Efeito de diferentes concentrações do óleo de nim no crescimento micelial de fungos entomopatogênicos e Trichoderma harzianum.

A associação de extratos de origem vegetal com fungos entomopatogênicos pode aumentar a eficiência do controle biológico de pragas e doenças de plantas e ainda reduzir custos e impactos ambientais. No presente trabalho foi avaliado o efeito do óleo de nim, incorporado ao meio BDA (Batata-dextrose-ágar) nas concentrações de 0, 1, 10, 100, 1.000, 10.000 e 100.000 μ/L, sobre o crescimento micelial de Metarhizium anisopliae, Beauveria bassiana, Trichoderma harzianum e Lecanicillium lecanii. O óleo de nim foi adicionado ao meio antes e após a esterilização em autoclave por 20 min. e 1 atm. Na parte central das placas de Petri contendo os meios foi transferido um disco de micélio dos agentes de biocontrole e mantidas em sala de incubação a 27 ± 2°C. O crescimento da colônia foi avaliado diariamente por seis dias, considerando dois sentidos do diâmetro. Cada placa correspondeu a uma repetição sendo cinco placas por tratamento. O óleo de nim adicionado antes e após autoclavagem em todas as concentrações testadas, estimularam o crescimento micelial de todos os agentes de biocontrole. Esta associação é de grande valia para o controle biológico, pode aumentar a eficiência destes fungos e reduzir impactos ambientais.

Trichoderma in Brazil: history, research, commercialization and perspectives.

2009

Control of Fusarium in chrysanthemum with sewage sludge, biofertilizer, hydrolyzed fish, chitosan and Trichoderma.

2009

Biological control of Sclerotinia sclerotiorum on beans in field by Trichoderma asperellum and Clonostachys rosea.

2009

Influência do óleo de nim no crescimento micelial, esporulação e viabilidade de Clonostachys rosea e Trichoderma harzianum.

2009

Redução do inóculo de Sclerotinia sclerotiorum por Trichoderma asperellum e Clonostachys rosea em feijão de inverno nas safras 2007 e 2008.

2009

Obtenção de Trichoderma stromaticum por fermentação em estado sólido, visando o controle da vassoura de bruxa.

2009

Ultrastructure of mycoparasitism of Trichoderma koningiopsis 4.24 and effects of its metabolic products on pathogenic fungi.

2009

Inibição do crescimento micelial de Sclerotinia sclerotiorum por filtrados de Trichoderma sp. e Clonostachys rosea.

Uma alternativa para controle do mofo-branco do feijão (Sclerotinia sclerotiorum, Ss) é o uso de agentes de controle biológico (ACB). Trichoderma sp. (Tr) e Clonostachys rosea (Cr) podem competir, parasitar e produzir metabólitos tóxicos contra fitopatógenos. Avaliou-se a capacidade de três isolados de Tr e um de Cr, previamente selecionados para o controle de Ss, em produzir metabólitos tóxicos contra o patógeno. Os ACB foram crescidos em caldo batata dextrose sob agitação (100 rpm) por sete dias. Após, o caldo foi filtrado a vácuo (membrana bacteriológica 0,22µm) e adicionou-se uma alíquota (2 ml) do filtrado ou de ADE (testemunha) a BDA fundente em placas de Petri. Após o resfriamento, adicionou-se no centro da placa um disco de micélio de Ss. As placas (cinco repetições/tratamento) foram incubadas a 20˚C ou 25˚C. Diariamente mediu-se o diâmetro das colônias até a testemunha atingir as bordas da placa. A 25˚C verificou-se significativa redução (Tukey, 5%) do crescimento micelial de Ss pelos filtrados dos três isolados de Tr (72 a 79%) e por Cr (50%). A 20ºC, apenas dois isolados de Tr inibiram Ss (45 a 65%). Os resultados indicam que os ACB podem atuar por antibiose, porém a eficiência é influenciada pelo ambiente.

Production of Recombinant Trichoderma Reesei Endoglucanase Protein CEL7B by Using Kluyveromyces Lactis

This research is about producing recombinant Trichoderma reesei endoglucanase Cel7B by using Kluyveromyces lactis, transformed with chromosomally integrated Cel7B cDNA, as a host cell (K. lactis Cel7B). Cel7B is one of the glycoside hydrolyze family of proteins that are produced by T. reesei. Cel7B together with other endoglucanases, exoglucanases, and â-glucosidases hydrolyze cellulose to glucose, which can then be fermented to biofuels or other value-added products. The research objective of this MS project is to examine favorable fermentation conditions for recombinant Cel7B enzyme production and improved activity. Production of enzyme on different types of media was examined, and the activity of the enzyme was measured by using different tools or procedures. The first condition tested for was using different concentrations of galactose as a carbon and energy source; however galactose also acts as a potent promoter of recombinant Cel7B expression in K. lactis Cel7B. The purpose of this method is to determine the relationship between production of enzyme with increasing sugar concentration. The second culture condition test was using different types of media: a complex medium-yeast extract, peptone, galactose (YPGal); a minimal medium-yeast nitrogen base (YNB) with galactose; and a minimal medium with supplement-yeast nitrogen base with casamino acid (YBC), a nitrogen source, with galactose. The third condition was using different types of reactors or fermenters: a small reactor (shake flask) and a larger automated bioreactor (BioFlo 3000 fermenter). The purpose of this method is to determine the quantity of the protein produced by using different environments of production. Different tools to determine the presence and activity of Cel7B enzyme were used. For the presence of enzyme, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used. Secondly, to detect enzyme activity, the carboxymethyl cellulose- 3,5-dinitrosalicylic acid (CMC- DNS) assay was employed. SDS-PAGE showed that the enzyme band was at 67 kDa, which is larger than native Cel7B (52 kDa.), likely due to over glycolylation during post-translational processing in K. lactis. For the different types of media used in our fermentation, recombinant Cel7B was produced from yeast extract peptone galactose (YPGal), and yeast nitrogen base with casamino acid (YBC), but was not produced and no activity was detected from yeast nitrogen base (YNB). This experiment concluded that the Cel7B production requires the amino acid resources as part of fermentation medium. In experiments where recombinant Cel7B net activity was measured at 1% galactose initial concentration in YPGal and YBC media, higher enzyme activity was detected for the complex medium YPGal. Higher activity of recombinant Cel7B was detected for flask culture in 2% galactose compared to 1% galactose for YBC medium. Two bioreactor experiments were conducted under these culture conditions at 30°C, pH 7.0, dissolved oxygen of 50% of saturation, and 250 rpm agitation (variable depending on DO control) K. lactis-Cel7B yeast growth curves were quite reproducible with maximum optical density (O.D) at 600 nm of between 7 and 8 (when factoring dilution of 10:1). Galactose was consumed rapidly during the first 15 hours of bioreactor culture and recombinant Cel7B started to appear in the culture at 10-15 hours and increased thereafter up to a maximum of between 0.9 and 1.6 mg/mL/hr in these experiments. These bioreactor enzyme activity results are much higher than comparable experiments conducted with flask-scale culture (0.5 mg/mL/hr). In order to achieve the highest recombinant Cel7B activity from batch culture of K. lactis-Cel7B, based on this research it is best to use a complex medium, 2% initial galactose concentration, and an automated bioreactor where good control of temperature, pH, and dissolved oxygen can be achieved.

Selection of Trichoderma spp. isolates to control the bean white-mold fungus Sclerotinia sclerotiorum in winter crops.

2007

Biological control of Sclerotinia sclerotiorum on beans in field by Trichoderma asperellum and Clonostachys rosea.

2008

Trichoderma in Brazil: history, research, commercialization and perspectives.

2008

Control of Fusarium in Chrysanthemum with sewage sludge, biofertilizer, hydrolyzed fish, chitosan and Trichoderma.

2008

Avaliação de isolados de Trichoderma sp. e Clonostachys rosea selecionados para o controle biológico de mofo-branco como agentes promotores de crescimento em feijoeiro.

2008