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En el cultivo de la vid, se reporta a Botrytis cinerea Pers como el hongo patógeno causante de lesiones cancrosas, necrosamiento de brotes y racimos de flores, y pudrición de frutos y semillas, que ocasionan grandes pérdidas económicas a los agricultores. El control de este patógeno se realiza hasta nuestros días con sustancias de origen químico: plaguicidas; los cuales son aplicados al follaje, a las semillas y al suelo, ocasionando problemas en el ambiente. En el presente trabajo, se colectaron 56 muestras de hojas de vid, infectadas con Botrytis cinerea Pers, en un viñedo del caserío La Huaca, en el distrito de San Benito, provincia de Contumaza, departamento de Cajamarca, durante el verano del 2006; de las cuales se aislaron e identificaron 12 cepas de Trichoderma viride. En la evaluación de la actividad antagonista de las 12 cepas de T. viride sobre B. cinerea mediante el método Precolonizado de Placas a 24° C y 28° C, se seleccionó a las cinco cepas: CH-13, CH-25, CH-30, CH-38 y CH-52 que presentaron los mayores porcentajes de área ocupada sobre el patógeno. Estas cepas presentaron actividad antagonista del tipo micoparasitismo, observándose enrollamiento y penetración de las hifas de las cepas de T. viride sobre las hifas de B. cinerea. De las cinco cepas seleccionadas de T. viride, la cepa CH-30 presentó el mayor porcentaje de área ocupada sobre B. cinerea a 24° C y 28° C con un 54.67% y 98% respectivamente. Por los resultados obtenidos, las cinco cepas nativas de Trichoderma viride pueden ser consideradas como una alternativa ecológica para su evaluación como agentes potenciales de control biológico del patógeno B. cinerea. / --- In the culturing of grapevine, there is reported Botrytis cinerea Pers as the fungus pathogenic causative of canker injuries, necrotizing of outbreaks and clusters of flowers, and rotting of fruits and seeds, which cause big economic losses to the farmers. The control of this pathogenic is realized actually by substances of chemical origin: pesticides; which are applied to the foliage, to the seeds and to the soil, causing environment problems. In this present work, there were collected 56 samples of leaves of grapevine, infected with Botrytis cinerea Pers, in a vineyard of The Huaca hamlet, in San Benito district, Contumaza province, Cajamarca department, during the summer of 2006; from these ones, there were isolated and identified 12 strains of Trichoderma viride. In the evaluation of the antagonist activity of these 12 strains of T. viride on B. cinerea by the method of Plates pre-colonized in 24° C and 28 °C, was selected five strains: CH-13, CH-25, CH-30, CH-38 and CH-52 that presented the major percentages of occupied area over the fungus pathogenic. These strains showed antagonist activity of mycoparasitism kind, being observed rolling and penetration of mycelium of T. viride strains on the mycelium of B. cinerea. From 5 T. viride strains selected, the CH-30 one showed the major percentage of occupied area on B. cinerea in 24° C and 28 °C with 54.67% and 98% respectively. For the results obtained, five native strains of Trichoderma viride can be considered as an ecological alternative for its evaluation as potential agents of biological control of the pathogenic B. cinerea.

La microflore des raisins est importante d’un point de vue technologique car elle conditionne en partie la qualité du vin. Or, la diversité des flores fongiques présentes sur baies de raisin ainsi que leur potentiel de contamination du produit final ne sont pas encore pleinement connus. Dans ce cadre, la caractérisation des flores fongiques cultivables présentes sur baies de raisin a été réalisée par PCR ITS-RFLP. 41 espèces de moisissures différentes sur les 43 étudiées appartenant à 11 genres différents ont été caractérisées de façon fiable. Seules les espèces Penicillium thomii et Penicillium glabrum ont présenté le même profil. Ainsi 96.3% des souches étudiées ont été caractérisées avec au maximum 4 enzymes de restriction et 41.5% des souches ont pu l’être avec seulement 2 enzymes de restriction. Ces résultats ont permis d’enrichir les bases de données, moyennement pourvues en séquences ITS caractéristiques de genres ou d’espèces de moisissures présentes sur baies de raisin. De plus, une étude exhaustive des moisissures présentes sur baies de raisin en Bourgogne a permis, par PCR ITS-RFLP, d’identifier 199 souches au niveau de l’espèce et ce quelque soit le genre. Penicillium spinulosum est l’espèce majoritaire isolée pour le millésime 2008 en Bourgogne. Parallèlement, la quantification de Botrytis cinerea, choisi comme micro-organisme modèle, a été réalisée par qPCR. La technique qPCR décrite dans ce travail présente (i) une bonne sensibilté avec une limite de détection de 6.4 pg d’ADN correspondant à 540 spores, (ii) l’originalité de travailler en échantillons naturellement contaminés et la fiabilité d’utiliser un standard interne. L’évaluation de l’efficacité de différentes stratégies de traitements anti-Botrytis a confirmé l’importance de la prophylaxie (effeuillage) dans la lutte contre Botrytis cinerea. / Microbial population of grapes is important from a technological point of view because it determines the quality of wine. But few studies have focused on fungal populations of grapes. A better knowledge of the fungal diversity on grapes, particularly as concerns species responsible for wine defects, may help efforts to control their development.We report the development of a PCR ITS-RFLP method as a fast and easy technique for identifying species of fungal genera present on grapes. By this methode, 41 different fungal species among 43 studied species belonging to 11 genera were characterized at the species level. Only P. thomii remained indistinguishable from P. glabrum. Using this PCR-ITS-RFLP, 96.3% strains tested could be differentiated to the species level with only four enzymes and 41.5% only with two enzymes. Moreover this work has contributed to the enriching of the database of fungal ITS sequences.Thus 199 isolated strain were on grapes in Burgundy vineyard were chacacterized at species level indepdantly of the genus by this method. P. spinolusum was the most frequently isolated species of Penicillium in Burgundy for 2008 vintage. Paralelly, the quantification of Botrytis cinerea, used as model, was developped by qPCR. The assay contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes, had high efficiency and the limit of detection was estimated to be 6.3 pg DNA (corresponding to 540 spores). This method was applied to assess the effects of various treatment strategies against Botrytis in the vineyard and demonstrates the importance of the prophylactic method.

En el cultivo de la vid, se reporta a Botrytis cinerea Pers como el hongo patógeno causante de lesiones cancrosas, necrosamiento de brotes y racimos de flores, y pudrición de frutos y semillas, que ocasionan grandes pérdidas económicas a los agricultores. El control de este patógeno se realiza hasta nuestros días con sustancias de origen químico: plaguicidas; los cuales son aplicados al follaje, a las semillas y al suelo, ocasionando problemas en el ambiente. En el presente trabajo, se colectaron 56 muestras de hojas de vid, infectadas con Botrytis cinerea Pers, en un viñedo del caserío La Huaca, en el distrito de San Benito, provincia de Contumaza, departamento de Cajamarca, durante el verano del 2006; de las cuales se aislaron e identificaron 12 cepas de Trichoderma viride. En la evaluación de la actividad antagonista de las 12 cepas de T. viride sobre B. cinerea mediante el método Precolonizado de Placas a 24° C y 28° C, se seleccionó a las cinco cepas: CH-13, CH-25, CH-30, CH-38 y CH-52 que presentaron los mayores porcentajes de área ocupada sobre el patógeno. Estas cepas presentaron actividad antagonista del tipo micoparasitismo, observándose enrollamiento y penetración de las hifas de las cepas de T. viride sobre las hifas de B. cinerea. De las cinco cepas seleccionadas de T. viride, la cepa CH-30 presentó el mayor porcentaje de área ocupada sobre B. cinerea a 24° C y 28° C con un 54.67% y 98% respectivamente. Por los resultados obtenidos, las cinco cepas nativas de Trichoderma viride pueden ser consideradas como una alternativa ecológica para su evaluación como agentes potenciales de control biológico del patógeno B. cinerea. / In the culturing of grapevine, there is reported Botrytis cinerea Pers as the fungus pathogenic causative of canker injuries, necrotizing of outbreaks and clusters of flowers, and rotting of fruits and seeds, which cause big economic losses to the farmers. The control of this pathogenic is realized actually by substances of chemical origin: pesticides; which are applied to the foliage, to the seeds and to the soil, causing environment problems. In this present work, there were collected 56 samples of leaves of grapevine, infected with Botrytis cinerea Pers, in a vineyard of The Huaca hamlet, in San Benito district, Contumaza province, Cajamarca department, during the summer of 2006; from these ones, there were isolated and identified 12 strains of Trichoderma viride. In the evaluation of the antagonist activity of these 12 strains of T. viride on B. cinerea by the method of Plates pre-colonized in 24° C and 28 °C, was selected five strains: CH-13, CH-25, CH-30, CH-38 and CH-52 that presented the major percentages of occupied area over the fungus pathogenic. These strains showed antagonist activity of mycoparasitism kind, being observed rolling and penetration of mycelium of T. viride strains on the mycelium of B. cinerea. From 5 T. viride strains selected, the CH-30 one showed the major percentage of occupied area on B. cinerea in 24° C and 28 °C with 54.67% and 98% respectively. For the results obtained, five native strains of Trichoderma viride can be considered as an ecological alternative for its evaluation as potential agents of biological control of the pathogenic B. cinerea.

<p>We hypothesize that South African medicinal plants contain compounds that can act in synergism with synthetic antifungal compounds. Four fungicides - Sporekill&trade / , Rovral&trade / , Terminator&trade / and Teldor&trade / at doses 0.1, 0.2, 0.4 and 0.8 mL L-1 and plant species Galenia africana, Elytropappus rhinocerotis and Tulbaghia violacea were tested aloneand in different combinations for their potency (efficacy) on radial growth inhibition of Botrytis cinerea strains on potato dextrose plates. Four doses of plant extract for each of the respective plant species were used. A total of 48 combinations were tested for each strain. Mixtures of plant extracts were far more effective in controlling strains compared to the individual components alone, representing significant levels of in vitro synergistic interactions. Combinations of these components represent an attractive future prospect for the development of new management strategies for controlling B. cinerea. Since the in vitro tests of these mixtures showed inhibitory activity, the mixtures were tested for activity in assays on Granny Smith apples. In vitro tests can be used to screen mixtures to obtain information on their inhibitory activity on a pathogen, however, the environmental conditions of the fruit and the ability of the pathogen to grow into the fruit cannot be simulated in vivo. A series of two-fold doses of medicinal plant extracts were combined with fungicides to conduct decay inhibition studies. The incidence of gray mold was significantly reduced by mixtures of plant extracts and fungicides. Under conditions similar to those in commercial storage, a drench treatment with G. africana and Rovral&trade / significantly (p=0.05) inhibit gray mold on the apples and was more effective than the plant extract and fungicide alone. The treatments exerted synergistic effects and were markedly better than the components applied alone. The wound colonization assay was used for optimal decay control. In a drench, much higher volumes of the treatments are used to ensure that the components of the suspension are deposited evenly over the entire fruit surface. Drenching of fruit to apply other chemicals is an established practise in the pome (fleshy) fruit industry, and simplifies the commercial application of the mixtures, as no additional infrastructure at commercial packing houses will be required. This approach not only makes it possible to reduce fungicide concentrations while maintaining adequate decay control, but also ensures a reduction of the chemical residue on the fruit.</p>

Gray mold, caused by Botrytis cinerea is the major cause of postharvest loss in strawberries. Detection of flower and fruit infections enables producers to make intelligent management decisions. A plate-trapped ELISA protocol using a Botrytis-specific monoclonal antibody (BC-12.CA4) was developed for the detection of Botrytis cinerea in strawberry flower receptacles and red fruits. Horseradish peroxidase, was chosen as enzyme conjugate because it gave lower background absorbance in disease-free samples. B. cinerea reference antigen (RAg) was isolated from strawberry. BC-12.CA4 was very sensitive to the RAg, detecting up to 6 mug/ml of RAg when mixed with strawberry extracts. The MAb did not show any reaction to Rhizopus sp., Mucor sp. and Penicillium sp. associated with strawberry. B. cinerea could be detected in receptacles two days after inoculation. Treatment of inoculated receptacles with paraquat speeded-up detection. Inoculated red fruit infection could be detected after three days of incubation. Disease in commercially-produced receptacles and red fruits were assessed visually and by ELISA. The ELISA detected B. cinerea in 95% of commercial flower samples, whereas the traditional visual method detected only 50 to 70%. No dramatic differences between methods were found for red fruits.

<p>We hypothesize that South African medicinal plants contain compounds that can act in synergism with synthetic antifungal compounds. Four fungicides - Sporekill&trade / , Rovral&trade / , Terminator&trade / and Teldor&trade / at doses 0.1, 0.2, 0.4 and 0.8 mL L-1 and plant species Galenia africana, Elytropappus rhinocerotis and Tulbaghia violacea were tested aloneand in different combinations for their potency (efficacy) on radial growth inhibition of Botrytis cinerea strains on potato dextrose plates. Four doses of plant extract for each of the respective plant species were used. A total of 48 combinations were tested for each strain. Mixtures of plant extracts were far more effective in controlling strains compared to the individual components alone, representing significant levels of in vitro synergistic interactions. Combinations of these components represent an attractive future prospect for the development of new management strategies for controlling B. cinerea. Since the in vitro tests of these mixtures showed inhibitory activity, the mixtures were tested for activity in assays on Granny Smith apples. In vitro tests can be used to screen mixtures to obtain information on their inhibitory activity on a pathogen, however, the environmental conditions of the fruit and the ability of the pathogen to grow into the fruit cannot be simulated in vivo. A series of two-fold doses of medicinal plant extracts were combined with fungicides to conduct decay inhibition studies. The incidence of gray mold was significantly reduced by mixtures of plant extracts and fungicides. Under conditions similar to those in commercial storage, a drench treatment with G. africana and Rovral&trade / significantly (p=0.05) inhibit gray mold on the apples and was more effective than the plant extract and fungicide alone. The treatments exerted synergistic effects and were markedly better than the components applied alone. The wound colonization assay was used for optimal decay control. In a drench, much higher volumes of the treatments are used to ensure that the components of the suspension are deposited evenly over the entire fruit surface. Drenching of fruit to apply other chemicals is an established practise in the pome (fleshy) fruit industry, and simplifies the commercial application of the mixtures, as no additional infrastructure at commercial packing houses will be required. This approach not only makes it possible to reduce fungicide concentrations while maintaining adequate decay control, but also ensures a reduction of the chemical residue on the fruit.</p>

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2004. / Härtill 4 uppsatser.

Orientador: Ben-Hur Mattiuz / Resumo: As doenças fúngicas são as principais causas de perdas na pós-colheita do tomate, e são controladas por manejo cultural e fungicidas aplicados na pré-colheita. O tratamento hiperbárico na pós-colheita é um métodos físico, com potencial uso comercial na redução da deteriorização por fungos. Neste trabalho, a eficiência das pressões hiperbáricas para controlar o desenvolvimento das podridões de Botrytis cinerea e Alternaria sp. in vitro e em tomates ‘Débora’ foram avaliadas, bem como a atividade das enzimas antioxidantes e as relacionadas a defesa do vegetal. Os experimentos foram realizados em um sistema de pressão hiperbárica do Laboratório de Tecnologia Pós-colheita da FCAV-UNESP, Câmpus de Jaboticabal, utilizando as pressões de 100 (controle), 200, 400, 600 e 800 kPa à temperatura de 23±1°C, durante 3 e 6 dias. No ensaio in vitro as placas com os fungos permaneceram nas pressões e depois foram incubadas até que a colônia do controle atingisse a borda. No experimento in vivo, tomates inoculados com os fungos foram mantidos por 3 e 6 dias sob o tratamento nas pressões, com posterior armazenamento por mais 2 e 4 dias em condição de ambiente (23 °C, 50% UR, 100 kPa) para simular a comercialização. Avaliou-se o crescimento micelial, a taxa de germinação dos conídios dos fungos, incidência e severidade das podridões nos frutos, o metabolismo antioxidante (teor de licopeno e polifenóis totais) e a atividade das enzimas relacionadas à defesa dos tomates (peroxidase (POD), fenilalan... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Fungal rots are one of the major causes of postharvest losses in tomatoes, and they are controlled by cultural management and fungicidal applied in preharvest. The postharvest hyperbaric treatment is a physical method that it have potential to be used in commercial for reducing fungal decay. In this work, the efficiency of hyperbaric pressures to control rot development of Botrytis cinerea and Alternaria sp. in vitro and ‘Debora’ tomatoes were evaluated, also the activity of antioxidant and defense-related enzymes.The experiments were carried out in a hyperbaric pressure system at FCAV-UNESP Postharvest Technology Laboratory, Campus de Jaboticabal, using the pressures of 100 (control), 200, 400, 600 and 800 kPa at a temperature of 23 ± 1 ° C for 3 and 6 days. In vitro assay the Petri dishes with the fungi were kept at pressures after they were incubated until the control colony reached the edge. In vivo assay, tomatoes inoculated with the fungus were kept at 3 and 6 days under the treatment pressures, after they were stored for more 2 and 4 days at ambient conditions (23 °C, 50% RH, 100 kPa) to simulate the commercialization. It was evaluated the mycelial growth, germination rate of fungal conidia, incidence and severity of fruit rot, antioxidant metabolism (lycopene content and total polyphenols) and activity of defence-related enzymes (eroxidase (POD), phenylalanine ammonia-lyase (PAL), β 1,3 glucanase (GLU), superoxide dismutase (SOD) and catalase (CAT)). The hyperbaric pr... (Complete abstract click electronic access below) / Doutor

Memoria para optar al Título Profesional de Ingeniero Agrónomo / Botrytis cinerea Pers. causa niveles importantes de pudrición, los que pueden superar el 10% en peras de exportación en Chile, principalmente en la variedad Packham’s Triumph. La enfermedad postcosecha más importante producida por B. cinerea en peras almacenadas en frío corresponde a la pudrición calicinal, cuyo control se realiza en floración, período en que el hongo produce infecciones latentes que se manifiestan en pudrición tras la madurez de la fruta. En años con primaveras húmedas se ha observado un aumento de la incidencia de la enfermedad, lo cual ha generado cuestionamientos acerca de los tratamientos de control e incertidumbre sobre la capacidad del patógeno de infectar los frutos durante su crecimiento. Los objetivos del presente estudio fueron determinar la ocurrencia de infecciones en campo durante el período de crecimiento de frutos bajo condiciones artificiales de alta humedad con y sin inoculación del patógeno, y determinar el nivel de infección en precosecha para evaluar la efectividad de las medidas de control realizadas por los productores en dos huertos comerciales de la zona central de Chile. En uno de los huertos estudiados se determinó que las inoculaciones lograron generar infecciones en el período de crecimiento de frutos, lo que permite concluir que el patógeno efectivamente puede infectar peras cv. Packham’s Triumph durante el crecimiento de los frutos. Por otro lado, el patógeno fue recuperado desde la cavidad calicinal de fruta no inoculada en crecimiento en ambos huertos, observándose también pudrición calicinal en postcosecha, lo que sería consecuencia de la inefectividad de las aplicaciones de fungicidas realizadas en floración por los productores. Este trabajo establece una base para la realización de estudios en campo en pudrición calicinal de perales en Chile. / Botrytis cinerea Pers. causes significant levels of postharvest decay that may be over 10% in pear fruit in Chile, especially in the cultivar Packham's Triumph. Calyx-end rot is the most important postharvest disease caused by B. cinerea in stored pears, and its control is achieved in the flowering period, when the fungus produces latent infections that manifest in fruit decay after ripening. It has been observed that there is an increased incidence of the disease in years with humid and rainy conditions during spring and summer, which has raised questions about control treatments and pathogen's ability to infect fruit during its growth under these conditions. The objectives of this study were to determine the occurrence of infections in the orchard during fruit growth under artificial high humidity conditions with and without pathogen inoculation, and to determine the level of preharvest infection in order to evaluate the efficacy of control measures taken by the growers in two commercial orchards of central Chile. In one of the studied orchards, inoculations resulted in infections during fruit growth, leading to the conclusion that the pathogen can effectively infect Packham's Triumph pears during this period. Moreover, the pathogen was isolated from the calyx cavity of non-inoculated fruit during the growth period, and postharvest calyx-end decay was also observed, which would indicate the inefficacy of fungicide applications performed by the growers. This work provides basic information for further field studies on calyx-end rot of pears in Chile.