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In Vitro” DE FUNGICIDAS PARA EL CONTROL DE HONGOS PATÓGENOS EN ESQUEJES DE CLAVEL DURANTE LA

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In Vitro” DE FUNGICIDAS PARA EL CONTROL DE HONGOS PATÓGENOS EN ESQUEJES DE CLAVEL DURANTE LA EVALUACIÒN “In Vitro” DE FUNGICIDAS PARA EL CONTROL DE HONGOS PATÓGENOS EN ESQUEJES DE CLAVEL DURANTE LA ETAPA DE ENRAIZAMIENTO DIANA PAOLA ALFONSO PULIDO EDNA ROCIO SANDOVAL SISA TRABAJO DE GRADO Presentado como requisito parcial para optar el titulo de MICROBIÓLOGO INDUSTRIAL PONTIFICIA UNIVERSIDAD JAVERIANA FACULTAD DE CIENCIAS CARRERA DE MICROBIOLOGÍA INDUSTRIAL Bogotá, D. C. Julio de 2008 1 NOTA DE ADVERTENCIA Artículo 23 de la Resolución N° 13 de Julio de 1946 “La Universidad no se hace responsable por los conceptos emitidos por sus alumnos en sus trabajos de tesis. Solo velará por que no se publique nada contrario al dogma y a la moral católica y por que las tesis no contengan ataques personales contra persona alguna, antes bien se vea en ellas el anhelo de buscar la verdad y la justicia”. 2 EVALUACIÒN “In Vitro” DE FUNGICIDAS PARA EL CONTROL DE HONGOS PATÓGENOS EN ESQUEJES DE CLAVEL DURANTE LA ETAPA DE ENRAIZAMIENTO DIANA PAOLA ALFONSO PULIDO EDNA ROCIO SANDOVAL SISA APROBADO Directora Codirectora Ma. CLEMENCIA F. DE LA ROTTA Ma. Fernanda Charry ING. AGR. M. Sc. FITOPATOLOGÍA Microbióloga industrial Asesora Zulma Arguelles Bacterióloga Floramérica C.I David Gómez M. Sc. Juan C. Hio Jurado Jurado 3 EVALUACIÒN “In Vitro” DE FUNGICIDAS PARA EL CONTROL DE HONGOS PATÓGENOS EN ESQUEJES DE CLAVEL DURANTE LA ETAPA DE ENRAIZAMIENTO DIANA PAOLA ALFONSO PULIDO EDNA ROCIO SANDOVAL SISA APROBADO Ingrid Schuler Ph.D Janeth Arias Palacios M. Sc. DECANO ACADEMICO DIRECTOR DE CARRERA 4 A Dios por haberme brindado paciencia y fortaleza para salir adelante obteniendo grandes triunfos. A mis padres Alberto y Rosaura por todo su amor, esfuerzo e incondicional apoyo durante toda mi vida. A mi hermana por su compañía y apoyo. A mi tia Blanca y Fernando por su constante apoyo. A “My princess” Valery por ser la luz de mis ojos y darle alegría a mi vida. A las personas que están ausentes pero desde donde se encuentran siempre fueron un apoyo espiritual. A todos mis amigos por su apoyo e incondicional amistad. A Edna por su compañía en los momentos difíciles. 5 A Dios por las personas que ha puesto en mi camino, por guiar cada instante de mi vida y permitir la finalización exitosa de este proyecto. A mi mama por fomentar en mí, valores de honestidad, superación y esfuerzo, si no fuera por su sacrificio no sería lo que soy y este sueño no la habría cumplido. A mi hermana, fiel amiga y consejera, por levantarme en los momentos más difíciles que hemos afrontado y por darme la alegría más grande de mi vida: mis sobrinitas. A mi papa, a pesar de la distancia en nuestros corazones por apoyarme de corazón y por los buenos recuerdos. A Andrés por darme momentos tan felices, por su amor incondicional y por sus palabras de aliento en los momentos indicados. A Paola por su humildad, comprensión y su gran amistad. 6 AGRADECIMIENTOS A la Dra María Clemencia Forero De La Rotta por guiarnos en el desarrollo de este proyecto y por su constante apoyo. A laboratorios Dr Calderon Asitencia Técnica Agrícola LTDA. por su colaboración en la financiación de este proyecto. Al laboratorio de sanidad vegetal de C.I. Floramerica LTDA. Por su contribución en el desarrollo de este proyecto. A María Fernanda Charry por estar siempre incondicionalmente en los momentos que más la necesitamos. A Zulma Arguelles Ramirez por su asesoría y apoyo en el desarrollo de este proyecto. A Andrés Libardo Rojas por su colaboración en el proyecto. A Liseth Castellanos por su aporte en el proyecto. 7 TABLA DE CONTENIDO PAGINA 1. Introducción 1 2. Marco Teórico 3 2.1. Generalidades del clavel 3 2.1.1.Suelos y clima 3 2.1.2.Taxonomía del clavel 3 2.2. Proceso de producción de las flores 4 2.3 Principales enfermedades en bancos de enraizamiento en clavel 9 2.3.1. Mancha foliar anillada Cladosporium echinulatatum. 11 2.3.2. Pudrición del tallo por Fusarium roseum 13 2.3.3. Pudrición de la corona Rhizoctonia solani 17 2.3.4. Alternariosis o tizón por Alternaría dianthi 21 2.4. Mecanismos de control de enfermedades 24 2.4.1. Control químico de fungicidas 24 2.4.2. Tipos de fungicidas 25 2.4.3. Mecanismos de acción 27 2.4.4. Resistencia a los fungicidas 27 2.5. Control biológico 29 2.5.1 Organismos útiles en control biológico 31 2.5.2. Mecanismos de control biológico 32 3. Justificación 35 4. Objetivos 36 4.1. Objetivos general 36 4.2. Objetivos específicos 36 5. Materiales y Métodos 37 5.1. Diseño experimental 37 8 5.2. Aislamiento de los hongos 38 5.3. Caracterización macroscópica y microscópica de los hongos 39 5.4. Evaluación del efecto fungicida y fungistático de los fungicidas 39 5.5. Inhibición de crecimiento micelial 40 5.5.1. Método de Bloques de Agar 41 5.5.2. Prueba de acción fungicida o fungistático 42 5.6. Evaluación por halos de inhibición 42 5.6.1. Métodos de pozos 42 5.6.2. Método de los discos de papel 43 5.7. Análisis estadístico 44 6. Resultados 45 6.1. Aislamiento y caracterización macroscópica y microscópica de los hongos. 45 6.2. Evaluación in vitro de fungicidas 48 6.2.1. Evaluación por halos de inhibición 48 6.2.2. Inhibición de crecimiento micelial 56 7. Discusión 64 8. Conclusiones 68 9. Recomendaciones 70 10. Referencias bibliográficas 71 11. Anexos 80 9 LISTA DE FIGURAS PÁG Figura 1: Síntomas en hojas de clavel causados por C. echinulatum 13 Figura 2. Ciclo de infección Fusarium sp en trigo 16 Figura 3. Síntomas ocasionados por F. roseum en hojas de clavel. 17 Figura 4. Ciclo de infección de Rhizoctonia sp 20 Figura 5. Síntomas causados por Rhizoctonia sp en raíces de clavel. 20 Figura 6. Ciclo de infección Alternaria sp 23 Figura 7. Síntomas causados por A. dianthi en hojas de clavel. 23 Figura 8. Características macroscópicas y microscópicas de A. dianthi. 45 Figura 9. Características macroscópicas y microscópicas de C. echinulatum. 46 Figura 10. Características macroscópicas y microscópicas de F. roseum 47 Figura 11. Características macroscópicas y microscópicas de R. solani 47 Figura 12. Halos de inhibición A. dianthi. 49 Figura 13. Halos de inhibición micelial A. dianthi. 50 Figura 14. Halos de inhibición C. echinulatum, Técnica de pozos Procloraz. 51 Figura 15. Halos de inhibición micelial obtenidos técnica de sensidiscos en la evaluación de fungicidas contra C. echinulatum. 51 Figura 16. Halos de inhibición F. roseum. para Trichoderma spp. 52 Figura 17. Halos de inhibición F. roseum. Técnica de sensidiscos para Carboxin + Captan 53 Figura 18. Halos de inhibición micelial obtenidos por la técnica de Sensidiscos para la evaluación de fungicidas contra F. roseum. 53 Figura 19. Halos de inhibición R. solani con Trichoderma spp 54 Figura 20. Halos de inhibición R. solani. con Metalaxil + Mancozeb. 55 Figura 21. Halos de inhibición micelial obtenidos técnica de sensidiscos para la evaluación de fungicidas contra R. solani. 55 10 Figura 22. Halos inhibición técnica sensidiscos R. solani. 56 Figura 23. Porcentajes inhibición técnica de bloques A. dianthi. 57 Figura 24. Técnica de Bloques A. dianthi frente a Trichoderma spp 58 Figura 25. Técnica de Bloques C. echinulatum frente a Trichoderma spp 58 Figura 26. Porcentajes inhibición técnica de bloques C. echinulatum 58 Figura 27. Técnica de Bloques para F. roseum frente a Trichoderma 60 Figura 28. Porcentajes de inhibición micelial contra F. roseum. 60 Figura 29. Técnica de Bloques para R. solani frente a Trichoderma spp. 61 Figura 30. Porcentajes inhibición técnica de bloques R. solani. 62 Figura No 31. Productos con mayor eficacia para el control de patógenos en clavel. 63 11 LISTA DE TABLAS PÁG Tabla No 1. Comparación entre sustratos utilizados en el enraizamiento: Ventajas y Desventajas. 7 Tabla 2. Principales agentes causales de enfermedades en cultivo de clavel a nivel mundial 10 Tabla 3. Fungicidas evaluados con sus respectivos tratamientos. 40 12 RESUMEN Los cultivos de flores día a día enfrentan problemas fitosanitarios donde se incluyen enfermedades causadas por hongos y bacterias los cuales disminuyen la calidad del producto final generando grandes perdidas económicas. El interés del proyecto fue evaluar tres tratamientos para 10 productos utilizados en el control de A. dianthi, C. echinulatum, F, roseum y R. solani hongos presentes en bancos de enraizamiento de clavel. Los productos se evaluaron mediante las técnicas in vitro de pozos, sensidiscos y trozos, lo que permite definir el mejor producto químico o biológico de control, expresado en porcentaje o halos de inhibición micelial, mediante un análisis de varianza ANOVA multifactorial. De acuerdo a los resultado obtenidos en la investigación el mejor producto fue Trichoderma spp, el cual mostró una eficaz inhibición de los hongos fitopatógenos evaluados, seguido de procloraz, carboxin + captan, etridiazol y metalaxil + mancozeb. Para los tratamientos se encontró diferencia estadísticamente significativa siendo el Tratamiento 1 el que mostró mas eficacia en la evaluación de los productos. Entre las técnicas de pozos y sensidiscos no se evidencio una diferencia estadísticamente significativa, pues los resultados para ambas técnicas fueron homogéneos. La acción fungistática o fungicida de los productos no se logro evaluar mediante la técnica de bloques pues se encontraron desventajas en el montaje de la técnica. 13 ABSTRACT The flowers’ crops day by day have problems phytosanitary, illnesses which include caused by fungi and bacteria which reduce the quality of the final product generating great economic losses. The interest of the project was to evaluate three treatments for 10 products used in controlling A. dianthi, C. echinulatum, F. roseum and R. solani fungus present in rooting banks of carnation.
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