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Fungi and Oomycetes represent the two most important groups of eukaryotic plant pathogens. Besides chemical and physical control of these pathogens, biological control is an approach enjoying increasingly more focus. One of the biological agents increasingly employed in biological control of plant pathogenic fungi is ironically the fungus Trichoderma, more specifically Trichoderma harzianum. Besides control of the fungal plant pathogens, another interesting aspect observed when plants are treated with Trichoderma harzianum are effects such as complete and even stand of plants, faster seed germination, increases in plant height and overall enhanced plant growth. Though there have been various studies on this effect, almost no research has yet been conducted to elucidate the mechanism by which these effects occur. In particular, effects such as faster seed germination suggest that Trichoderma harzianum produces a metabolite that may mimic the plant growth hormone gibberellic acid. Through an evaluation of the various metabolites produced by Trichoderma harzianum; gliotoxin seemed structurally most similar to gibberellic acid. To verify that gliotoxin can indeed serve as an analogue for gibberellic acid and elicit similar physiological responses in plants, a two–pronged approach was followed. Firstly, molecular similarity evaluation through common pharmacophore evaluation was conducted, followed by docking simulations into the recently discovered receptor for gibberellic acid. Common pharmacophore evaluation between gibberellic acid and gliotoxin showed successful alignment of gliotoxin into the gibberellic acid based pharmacophore space. Furthermore, docking simulations further strengthened this by the similarity in docking scores calculated and the similar poses of the ligands (gliotoxin and gibberellic acid) in the receptor space. However, similarity in pharmacophore alignment and docking simulation results only suggest that gliotoxin should be able to occupy the receptor space, but it is not a guarantee that similar physiological responses will be elicited. In the second part of the project, the ability of gliotoxin to elicit similar physiological responses in plants to gibberellic acid was investigated. For this, a–amylase induction; plant emergence and height; and chlorophyll fluorescence were compared for both gliotoxin and gibberellic acid treatments. In terms of a–amylase induction, gliotoxin was able to induce production of the enzyme as visualised by starch–containing native gel electrophoresis (zymograms). Gliotoxin induced the strongest response at a 10–6 M dilution which is typically the range expected for hormones in biological systems in de–embryonated seeds of Phaseolus vulgaris. Gibberellic acid was able to induce the strongest response at a 10–7 M dilution. In essence, similar physiological responses were observed. In terms of plant emergence and plant height, treatment with gliotoxin or gibberellic acid resulted in plant emergence a day earlier than the untreated control. However, even though there were slight differences in plant height favouring the gliotoxin or gibberellic acid treated plants, the differences were not statistically significant. Thus, in this regard similar responses were again observed for both gliotoxin and gibberellic acid treatments. In the final evaluation the effect of gliotoxin and gibberellic acid treatments on the chlorophyll fluorescence of mature plants was investigated. Overall, both gliotoxin and gibberellic acid elicited beneficial effects on plant vitality, expressed through PI(Abs) with the gliotoxin treatment performing better than the equivalent gibberellic acid treatment. Overall, the physiological tests demonstrated that gliotoxin can indeed elicit similar positive physiological responses to gibberellic acid in Phaseolus vulgaris. Furthermore the test used in this project can serve as a standard evaluation bench for screening for gibberellic acid analogues on a laboratory scale before larger scale field trials are considered. / Thesis (Ph.D. (Microbiology))--North-West University, Potchefstroom Campus, 2012.

Fungi and Oomycetes represent the two most important groups of eukaryotic plant pathogens. Besides chemical and physical control of these pathogens, biological control is an approach enjoying increasingly more focus. One of the biological agents increasingly employed in biological control of plant pathogenic fungi is ironically the fungus Trichoderma, more specifically Trichoderma harzianum. Besides control of the fungal plant pathogens, another interesting aspect observed when plants are treated with Trichoderma harzianum are effects such as complete and even stand of plants, faster seed germination, increases in plant height and overall enhanced plant growth. Though there have been various studies on this effect, almost no research has yet been conducted to elucidate the mechanism by which these effects occur. In particular, effects such as faster seed germination suggest that Trichoderma harzianum produces a metabolite that may mimic the plant growth hormone gibberellic acid. Through an evaluation of the various metabolites produced by Trichoderma harzianum; gliotoxin seemed structurally most similar to gibberellic acid. To verify that gliotoxin can indeed serve as an analogue for gibberellic acid and elicit similar physiological responses in plants, a two–pronged approach was followed. Firstly, molecular similarity evaluation through common pharmacophore evaluation was conducted, followed by docking simulations into the recently discovered receptor for gibberellic acid. Common pharmacophore evaluation between gibberellic acid and gliotoxin showed successful alignment of gliotoxin into the gibberellic acid based pharmacophore space. Furthermore, docking simulations further strengthened this by the similarity in docking scores calculated and the similar poses of the ligands (gliotoxin and gibberellic acid) in the receptor space. However, similarity in pharmacophore alignment and docking simulation results only suggest that gliotoxin should be able to occupy the receptor space, but it is not a guarantee that similar physiological responses will be elicited. In the second part of the project, the ability of gliotoxin to elicit similar physiological responses in plants to gibberellic acid was investigated. For this, a–amylase induction; plant emergence and height; and chlorophyll fluorescence were compared for both gliotoxin and gibberellic acid treatments. In terms of a–amylase induction, gliotoxin was able to induce production of the enzyme as visualised by starch–containing native gel electrophoresis (zymograms). Gliotoxin induced the strongest response at a 10–6 M dilution which is typically the range expected for hormones in biological systems in de–embryonated seeds of Phaseolus vulgaris. Gibberellic acid was able to induce the strongest response at a 10–7 M dilution. In essence, similar physiological responses were observed. In terms of plant emergence and plant height, treatment with gliotoxin or gibberellic acid resulted in plant emergence a day earlier than the untreated control. However, even though there were slight differences in plant height favouring the gliotoxin or gibberellic acid treated plants, the differences were not statistically significant. Thus, in this regard similar responses were again observed for both gliotoxin and gibberellic acid treatments. In the final evaluation the effect of gliotoxin and gibberellic acid treatments on the chlorophyll fluorescence of mature plants was investigated. Overall, both gliotoxin and gibberellic acid elicited beneficial effects on plant vitality, expressed through PI(Abs) with the gliotoxin treatment performing better than the equivalent gibberellic acid treatment. Overall, the physiological tests demonstrated that gliotoxin can indeed elicit similar positive physiological responses to gibberellic acid in Phaseolus vulgaris. Furthermore the test used in this project can serve as a standard evaluation bench for screening for gibberellic acid analogues on a laboratory scale before larger scale field trials are considered. / Thesis (Ph.D. (Microbiology))--North-West University, Potchefstroom Campus, 2012.

O presente trabalho tem como objetivo estudar alternativas de controle químico e biológico de isolados de Pythium para aplicação na formação de mudas de tabaco. Estudou-se por meio de bioensaios a compatibilidade de duas cepas de Trichoderma harzianum (ESALQ 1306 e 1316) com sete fungicidas. Os mesmos fungicidas foram avaliados quanto à capacidade de inibição de quatro isolados de Pythium spp. A atividade antagônica das cepas de T. harzianum foram testadas in vitro, em três momentos de inoculação a 23 e 28ºC para quatro isolados de Pythium spp. A eficiência de controle in vivo de Pythium sp., bem como germinação, massa seca e mudas aptas foram avaliadas em condições de casa de vegetação, com os fungicidas químicos Ridomil Gold MZ e Infinito, e biológicos Trichodermil 1306 e o produto experimental Trichodermil 1316. As cepas de T. harzianum apresentaram compatibilidade com todos os fungicidas testados. Todos os isolados de Pythium foram inibidos pelos fungicidas Censor, Amistar WG 500, Infinito e Ridomil Gold MZ em ensaios. A cepa ESALQ 1316 apresentou maior eficiência de inibição dos isolados de Pyhtium, em cultivo pareado, sendo 100% eficiente, independente do momento de inoculação, após 14 dias. Isoladamente o Trichodermil cepa ESALQ 1306 foi mais eficiente que o produto experimental Trichodermil cepa ESALQ 1316 no controle de Pythium sp. em mudas de tabaco. A combinação do Ridomil Gold MZ ou Infinito com o Trichodermil 1316 apresentou maior eficiência de controle de Pythium sp. quando comparada ao agente de biocontrole isoladamente em mudas de tabaco. / The present work aims to study alternatives for chemical and biological control of Pythium isolates for application in tobacco seedlings. It was studied by bioassays the compatibility of two strains of Trichoderma harzianum (ESALQ 1306 and 1316) with seven fungicides. The same fungicides were evaluated for their ability to inhibit four isolates of Pythium sp. The antagonistic activity of strains of T. harzianum were tested in vitro on three different occasions inoculation at 23 and 28 º C for four isolates of Pythium sp. The control efficiency in vivo of Pythium sp. and germination, seedling dry weight and suitable conditions were evaluated in a greenhouse with chemical fungicides Ridomil Gold MZ and Infinito, and biological products, Trichodermil 1306 and Trichodermil 1316. The strains of T. harzianum showed compatibility with all fungicides tested. All the isolates of Pythium were inhibited by fungicides Censor, Amistar WG 500, Infinito and Ridomil Gold MZ in trials. The strain ESALQ 1316 was more efficient inhibition of isolates Pyhtium in paired culture, being 100% effective, regardless of time of inoculation, after 14 days. On average Trichodermil 1306 was more efficient than the experimental product Trichodermil 1316 in the control of Pythium sp. in tobacco seedlings. The combination of Ridomil Gold MZ or Infinito with Trichodermil 1316 was more effective control of Pythium sp. compared with the biological control agent alone in tobacco seedlings.

Memoria para optar al Título Profesional de: Ingeniero Agrónomo / Se evaluó la capacidad bioantagónica in vitro de diferentes bacterias (Bacillus sp. cepa 1; Paenibacillus sp. cepas 1 y 2) y hongos (cepas 24-A y 24-B de Clonostachys rosea; cepas Th 650, Th 12A 10.1 y ThF 2-1 de Trichoderma harzianum) y los productos comerciales Actinovate® (Streptomyces lydicus WYEC 108) y Trichonativa® (Trichoderma virens, Trichoderma harzianum y Trichoderma parceramosum) en el control de 13 cepas de basidiomycotas recuperadas desde plantas de vid con síntomas de enrollamiento clorótico y degradación de la madera, provenientes de 3 regiones de Chile (IV Región de Coquimbo, V Región de Valparaíso y RM de Chile). Se caracterizaron las diferentes cepas de hongos aislados respecto a temperatura (19, 22, 25 y 28ºC) y pH (5, 6, 7 y 8). La capacidad bioantagónica se determinó a través de pruebas de antagonismo directo realizado mediante cultivos duales y antagonismo por metabolitos difusibles. En los resultados de caracterización de las cepas recuperadas, se observó que el mayor crecimiento miceliar se obtuvo a 28ºC a un pH 6 del medio, determinándose ambos valores como los óptimos para el desarrollo de los aislados investigados. Acorde con los resultados obtenidos en antagonismo directo, todos los bioantagonistas demostraron tener un cierto grado de capacidad antagónica sobre las cepas. Mediante la inhibición del crecimiento miceliar, la bacteria que presentó un mayor efecto fue la cepa 1 de Paenibacillus sp., con un máximo de 63,5% de inhibición. En el caso de los hongos fueron todas las cepas de T. harzianum utilizadas, oscilando el máximo porcentaje de control entre un 82,7% para la cepa Th 650, un 84,6% para Th 12A 10.1 y 87,7% para la cepa ThF 2-1. En relación a los productos comerciales, Trichonativa® logró un 61,5% de inhibición miceliar, mientras que Actinovate® logró un 100% de inhibición, siendo por lo tanto S. lydicus WYEC108 el antagonista que ejerció el mayor efecto. Los resultados obtenidos en el ensayo de antagonismo mediante metabolitos difusibles ayudaron a corroborar el tipo de mecanismo de biocontrol que ejercieron los antagonistas. Las bacterias utilizadas para este ensayo reflejaron que básicamente son los metabolitos difusibles que producen, los que ejercen la inhibición de crecimiento miceliar de las cepas de basidiomycotas, obteniéndose con ambas cepas 1, de Bacillus sp. y Paenibacillus sp., valores en promedio superiores a 70%; en relación al producto comercial Actinovate®, se obtuvo una completa inhibición miceliar. Con respecto a los hongos C. rosea 24-B y T. harzianum ThF 2-1, estos mostraron una baja inhibición mediante la producción de metabolitos difusibles, obteniéndose valores en promedio que no superaron el 14% de inhibición de crecimiento miceliar, lo que indica que poseen otro tipo de mecanismos como en el caso de las cepas de T. harzianum. Para el producto comercial Trichonativa®, se obtuvo en promedio un 16,7% de inhibición miceliar, siendo éste el mayor dentro de los hongos. Los resultados obtenidos señalan a S. lydicus WYEC108 (Actinovate®) como al bioantagonista de mayor efecto. / The in vitro bioantagonistic capacity of different bacteria (Bacillus sp. strain 1, Paenibacillus sp. strains 1 and 2) and fungi (24-A and 24-B strains of Clonostachys rosea, and Th 650, Th 12A 10.1 and ThF 2-1 strains of Thrichoderma harzianum) and of the biofungicides Actinovate ® (Streptomyces lydicus WYEC 108) and Trichonativa ® (Trichoderma virens, Trichoderma harzianum and Trichoderma parceramosum) was evaluated in the control of 13 basidiomycota strains recovered from grapevines grown in three regions of Chile (IV Region of Coquimbo, V Region of Valparaiso and Metropolitan Region). The vines showed wood degradation and chlorotic leafroll symptoms. Different isolated fungus strains were characterized with respect to temperature (19, 22, 25, and 28ºC) and pH (5, 6, 7, and 8). The bioantagonistic capacity was determined by tests of direct antagonism with dual cultures and by diffusible metabolite antagonism. The results of recovered strain characterization showed that mycelial growth was highest with the culture medium at 28ºC and pH 6, indicating both values as optimum for the development of the investigated isolates. According to the direct antagonism results, all bioantagonists demonstrated to have some degree of antagonistic capacity over recovered strains. Through mycelial growth inhibition, the bacterium that showed a better effect was Paenibacillus sp. strain 1, with a maximum of 63.5% of inhibition. In the case of fungi, all the used T. harzianum strains had excellent results with the highest control percentages being 82.7% for the Th 650 strain, 84.6% for the Th 12A 10.1 strain and 87.7% for ThF 2-1 strain. In relation to the biofungicides tested, Trichonativa ® achieved a maximum of 61.5% of mycelial inhibition, while Actinovate ® showed 100% of inhibition of the 13 basidomycota strains. The results obtained in the antagonism trial by diffusible metabolite helped to corroborate the type of biocontrol mechanism exerted by the antagonists. The bacteria used for this trial basically showed that the diffusible metabolites that they produce are the ones which exert mycelial growth inhibition of basidiomycota strains, with both Bacillus sp. and Paenibacillus sp. strains 1 showing average values over 70%. With respect to Actinovate ®, complete mycelial growth inhibition was obtained with this biofungicide. Regarding fungi C. rosea 24-B and T. harzianum ThF 2-1, these showed low inhibition by diffusible metabolite production with average values not exceeding 14% of mycelial growth inhibition, which indicates that they have other types of mechanisms as in the case of T. harzianum strains. With Trichonativa ®, an average of 16.7% of mycelial growth inhibition was obtained, this biofungicide being the higher within the fungi used. The obtained results showed S. lydicus WYEC108 (Actinovate®) as the bioantagonist with higher effect.

Made available in DSpace on 2016-06-02T20:21:35Z (GMT). No. of bitstreams: 1 6150.pdf: 1801179 bytes, checksum: d2366a6c320c54df184a270c81c7c653 (MD5) Previous issue date: 2014-02-27 / The present situation of energy and environmental setting in the world has favored the development of alternative energy sources, and has emerged as an option the use of biofuels, such as bioethanol. Besides ethanol produced from sugarcane, there is the alternative of to use plant biomass to get called second-generation ethanol. In Brazil, the major lignocellulosic material used to produce bioethanol is sugarcane bagasse, and this ethanol is obtained by fermentation of the hydrolyzate of the plant cell wall polysaccharides, this hydrolysis is performed normally by enzymes cellulolytic. Microorganisms, such as fungi of the genus Trichoderma, which are filamentous fungi present in the soil, have a variety of these enzymes that convert cellulose into glucose. In this context, the aim of this study was the recombinant production of Trichoderma harzianum IOC-3844 endoglucanase IV and its biochemical characterization. This Endo IV, which presents high similarity with Endo IV of other Thichoderma species, was expressed using the heterologous expression system of the yeast Pichia pastoris, and after be purified, assays were performed to test their activity. The recombinant enzyme hasn t show sign of degradation in any of the substrates used and can t be confirmed its hydrolytic action. However, the literature has been showed that these enzymes have capacity to increase the breaking of lignocellulose by cellulases (or a mix of them). Based on these data, a test was performed to evaluate the ability of endo IV supplemental activity in the cocktail Acellerase 1500 (Genencor) in the hydrolysis of biomass. In the experiment, copper was added as an electron donor and despite the increase in the hydrolysis of sugarcane bagasse the results showed that copper alone improves the cocktail performance. A circular dichroism analysis was performed and showed that the recombinant Endo IV is correctly coiled. The study showed that the enzyme produced at the laboratory has a high level of expression, but its activity couldn t be detected. The intention now is to perform tests with cocktails that don t contain expansins and isolated enzymes in order to verify that the Endo IV has effect upon them. / A atual situação do cenário energético e ambiental do mundo tem favorecido o desenvolvimento de fontes alternativas de energia, e como opção tem surgido a utilização de biocombustíveis, como o bioetanol. Além do etanol produzido a partir do caldo de cana, há a alternativa de utilizar a biomassa vegetal para obter o chamado etanol de segunda geração. No Brasil o principal material lignocelulósico utilizado para produção de bioetanol é o bagaço de cana-de-açúcar, e este etanol é obtido por meio da fermentação do hidrolisado dos polissacarídeos da parede celular dos vegetais, sendo esta hidrólise, realizada, normalmente, por enzimas celulolíticas. Microrganismos, como os fungos do gênero Trichoderma, que são fungos filamentosos presentes no solo, possuem uma variedade dessas enzimas capazes de converter a celulose em glicose. Nesse contexto, o objetivo deste trabalho foi a produção recombinante da enzima endoglucanase IV de Trichoderma harzianum IOC-3844 e sua caracterização bioquímica. Esta Endo IV, que apresenta alta similaridade com as Endo IV de outras espécies de Thichoderma, foi expressa utilizando-se o sistema de expressão heteróloga da levedura Pichia pastoris e, após ser purificada, foram realizados ensaios para testar sua atividade. A enzima recombinante não demonstrou sinal de degradação em nenhum dos substratos utilizados, não podendo ser comprovada sua ação hidrolítica. Porém, na literatura consta que essas enzimas têm a capacidade de aumentar a quebra de lignocelulose por celulases (ou uma mistura delas). Com base nesses dados, um teste foi realizado para avaliar a capacidade da endo IV de suplementar a atividade do coquetel Acellerase 1500 (Genencor) na hidrólise de biomassa. No experimento, foi adicionado cobre como doador de elétrons e apesar do aumento na hidrólise do bagaço-de-cana os resultados mostraram que o cobre sozinho melhora a performance do coquetel. Uma análise de dicroísmo circular foi realizada e revelou que a Endo IV recombinante se encontra corretamente enovelada. O estudo mostrou que a enzima produzida em laboratório possui alto nível de expressão, porém sua atividade não pôde ser constatada. Pretende-se agora realizar testes com coquetéis que não contêm expansinas e com enzimas isoladas, a fim de verificar se a Endo IV possui efeito sobre elas.

O presente trabalho tem como objetivo estudar alternativas de controle químico e biológico de isolados de Pythium para aplicação na formação de mudas de tabaco. Estudou-se por meio de bioensaios a compatibilidade de duas cepas de Trichoderma harzianum (ESALQ 1306 e 1316) com sete fungicidas. Os mesmos fungicidas foram avaliados quanto à capacidade de inibição de quatro isolados de Pythium spp. A atividade antagônica das cepas de T. harzianum foram testadas in vitro, em três momentos de inoculação a 23 e 28ºC para quatro isolados de Pythium spp. A eficiência de controle in vivo de Pythium sp., bem como germinação, massa seca e mudas aptas foram avaliadas em condições de casa de vegetação, com os fungicidas químicos Ridomil Gold MZ e Infinito, e biológicos Trichodermil 1306 e o produto experimental Trichodermil 1316. As cepas de T. harzianum apresentaram compatibilidade com todos os fungicidas testados. Todos os isolados de Pythium foram inibidos pelos fungicidas Censor, Amistar WG 500, Infinito e Ridomil Gold MZ em ensaios. A cepa ESALQ 1316 apresentou maior eficiência de inibição dos isolados de Pyhtium, em cultivo pareado, sendo 100% eficiente, independente do momento de inoculação, após 14 dias. Isoladamente o Trichodermil cepa ESALQ 1306 foi mais eficiente que o produto experimental Trichodermil cepa ESALQ 1316 no controle de Pythium sp. em mudas de tabaco. A combinação do Ridomil Gold MZ ou Infinito com o Trichodermil 1316 apresentou maior eficiência de controle de Pythium sp. quando comparada ao agente de biocontrole isoladamente em mudas de tabaco. / The present work aims to study alternatives for chemical and biological control of Pythium isolates for application in tobacco seedlings. It was studied by bioassays the compatibility of two strains of Trichoderma harzianum (ESALQ 1306 and 1316) with seven fungicides. The same fungicides were evaluated for their ability to inhibit four isolates of Pythium sp. The antagonistic activity of strains of T. harzianum were tested in vitro on three different occasions inoculation at 23 and 28 º C for four isolates of Pythium sp. The control efficiency in vivo of Pythium sp. and germination, seedling dry weight and suitable conditions were evaluated in a greenhouse with chemical fungicides Ridomil Gold MZ and Infinito, and biological products, Trichodermil 1306 and Trichodermil 1316. The strains of T. harzianum showed compatibility with all fungicides tested. All the isolates of Pythium were inhibited by fungicides Censor, Amistar WG 500, Infinito and Ridomil Gold MZ in trials. The strain ESALQ 1316 was more efficient inhibition of isolates Pyhtium in paired culture, being 100% effective, regardless of time of inoculation, after 14 days. On average Trichodermil 1306 was more efficient than the experimental product Trichodermil 1316 in the control of Pythium sp. in tobacco seedlings. The combination of Ridomil Gold MZ or Infinito with Trichodermil 1316 was more effective control of Pythium sp. compared with the biological control agent alone in tobacco seedlings.

This paper deals with evaluation of presence of pathogenic fungi in oats, in 4 strains and their variants, ways of treatment during growing season. As well, presence rate of studied phytopathogenic fungi, strategy of loss estimate and evalution of intensity of disease are determined. Furhermore, infection of pathogenes of Fusarium fungi is monitored, including evaluation of yield-forming elements. The purpose of in vitro experiments is to observe surficial microflora on oat seed.

O presente trabalho tem como objetivo estudar alternativas de controle químico e biológico de isolados de Pythium para aplicação na formação de mudas de tabaco. Estudou-se por meio de bioensaios a compatibilidade de duas cepas de Trichoderma harzianum (ESALQ 1306 e 1316) com sete fungicidas. Os mesmos fungicidas foram avaliados quanto à capacidade de inibição de quatro isolados de Pythium spp. A atividade antagônica das cepas de T. harzianum foram testadas in vitro, em três momentos de inoculação a 23 e 28ºC para quatro isolados de Pythium spp. A eficiência de controle in vivo de Pythium sp., bem como germinação, massa seca e mudas aptas foram avaliadas em condições de casa de vegetação, com os fungicidas químicos Ridomil Gold MZ e Infinito, e biológicos Trichodermil 1306 e o produto experimental Trichodermil 1316. As cepas de T. harzianum apresentaram compatibilidade com todos os fungicidas testados. Todos os isolados de Pythium foram inibidos pelos fungicidas Censor, Amistar WG 500, Infinito e Ridomil Gold MZ em ensaios. A cepa ESALQ 1316 apresentou maior eficiência de inibição dos isolados de Pyhtium, em cultivo pareado, sendo 100% eficiente, independente do momento de inoculação, após 14 dias. Isoladamente o Trichodermil cepa ESALQ 1306 foi mais eficiente que o produto experimental Trichodermil cepa ESALQ 1316 no controle de Pythium sp. em mudas de tabaco. A combinação do Ridomil Gold MZ ou Infinito com o Trichodermil 1316 apresentou maior eficiência de controle de Pythium sp. quando comparada ao agente de biocontrole isoladamente em mudas de tabaco. / The present work aims to study alternatives for chemical and biological control of Pythium isolates for application in tobacco seedlings. It was studied by bioassays the compatibility of two strains of Trichoderma harzianum (ESALQ 1306 and 1316) with seven fungicides. The same fungicides were evaluated for their ability to inhibit four isolates of Pythium sp. The antagonistic activity of strains of T. harzianum were tested in vitro on three different occasions inoculation at 23 and 28 º C for four isolates of Pythium sp. The control efficiency in vivo of Pythium sp. and germination, seedling dry weight and suitable conditions were evaluated in a greenhouse with chemical fungicides Ridomil Gold MZ and Infinito, and biological products, Trichodermil 1306 and Trichodermil 1316. The strains of T. harzianum showed compatibility with all fungicides tested. All the isolates of Pythium were inhibited by fungicides Censor, Amistar WG 500, Infinito and Ridomil Gold MZ in trials. The strain ESALQ 1316 was more efficient inhibition of isolates Pyhtium in paired culture, being 100% effective, regardless of time of inoculation, after 14 days. On average Trichodermil 1306 was more efficient than the experimental product Trichodermil 1316 in the control of Pythium sp. in tobacco seedlings. The combination of Ridomil Gold MZ or Infinito with Trichodermil 1316 was more effective control of Pythium sp. compared with the biological control agent alone in tobacco seedlings.

Memoria para optar al título profesional de: Ingeniero Agrónomo Mención: Sanidad Vegetal