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Evaluación in Vitro De La Capacidad Antagónica De Hongos Endófitos Aislados De Moringa Oleífera (L) En El Control De Pestalotia Palmarum

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Evaluación in Vitro De La Capacidad Antagónica De Hongos Endófitos Aislados De Moringa Oleífera (L) En El Control De Pestalotia Palmarum EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 1 EVALUACIÓN IN VITRO DE LA CAPACIDAD ANTAGÓNICA DE HONGOS ENDÓFITOS AISLADOS DE MORINGA OLEÍFERA (L) EN EL CONTROL DE PESTALOTIA PALMARUM (Q). Samia Hasay Montoya Sánchez UNIVERSIDAD DE SANTANDER UDES FACULTAD DE CIENCIAS EXACTAS, NATURALES Y AGROPECUARIAS BUCARAMANGA 2019 EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 2 EVALUACIÓN IN VITRO DE LA CAPACIDAD ANTAGÓNICA DE HONGOS ENDÓFITOS AISLADOS DE MORINGA OLEÍFERA (L) EN EL CONTROL DE PESTALOTIA PALMARUM (Q). Samia Hasay Montoya Sánchez Trabajo de grado presentado como requisito para optar al título de Microbióloga Industrial Director Beatriz Elena Guerra Sierra PhD UNIVERSIDAD DE SANTANDER UDES FACULTAD DE CIENCIAS EXACTAS, NATURALES Y AGROPECUARIAS BUCARAMANGA 2019 EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 3 EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 4 “A Dios por permitirme haber llegado hasta este momento tan importante de mi formación profesional. A mi madre, por ser el pilar más importante, por ser mi gran amiga y compañera, por su amor, paciencia y esfuerzo. Gracias madre por tu apoyo incondicional eternamente agradecida contigo. TE AMO MONA” EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 5 Agradecimientos Primeramente, agradecerle a Dios por ser mi guía y acompañarme en el transcurso de mi vida, brindándome paciencia y sabiduría para culminar con éxito mis metas propuestas. A mi madre por ser mi guía, por ser mi apoyo incondicional, por su gran esfuerzo. A mis abuelos por estar siempre a mi lado por colaborarme siempre que fuera necesario. A mis tíos, tías, primos y primas por acompañarme en este proceso. A mi Tutora Beatriz Elena Guerra Sierra, por su confianza puesta en mí, por su apoyo para con mi trabajo. Gracias por los conocimientos brindados. A Adriana Sandoval por su gran entrega, colaboración y seguimiento en cada uno de los momentos de mi trabajo. Mi agradecimiento a todos, familia y amigos que de una u otra manera me brindaron su colaboración y se involucraron en este proyecto. EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 6 Contenido Pág. Resumen ....................................................................................................................................... 12 Abstract ........................................................................................................................................ 13 Introducción ................................................................................................................................ 14 1 Presentación del trabajo de grado ..................................................................................... 17 1.1 Planteamiento del Problema ....................................................................................... 17 1.2 Pregunta de Investigación ........................................................................................... 19 1.3 Hipótesis ..................................................................................................................... 19 1.4 Objetivos ..................................................................................................................... 19 1.4.1 Objetivo general .................................................................................................. 19 1.4.2 Objetivos específicos........................................................................................... 19 1.5 Justificación .................................................................................................................... 20 2 Marco teórico ....................................................................................................................... 22 2.1 Palma aceitera ............................................................................................................. 22 2.2 Insecto facilitador de la Pestaloptiosis ........................................................................ 24 2.3 Pestalotia palmarum................................................................................................... 26 2.3.1 Generalidades. ..................................................................................................... 27 2.4 Pestalotiopsis .............................................................................................................. 30 2.4.1 Controles Químicos. ............................................................................................ 30 2.5 Moringa oleífera Lam ................................................................................................ 32 2.6 Hongos endófitos ........................................................................................................ 34 2.6.1 Metabolitos secundarios. ..................................................................................... 36 2.6.2 Mecanismos de Acción. Los mecanismos de acción del antagonismo dual se traducen en competencia por espacio y alimento, antibiosis, parasitismo y sinergismo. ...... 37 3 Metodología .......................................................................................................................... 39 3.1 Población y muestra.................................................................................................... 39 3.2 Sitio de estudio ........................................................................................................... 39 3.3 Manejo y tratamiento de las muestras ........................................................................ 39 EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 7 3.4 Siembra de los fragmentos del material vegetal ......................................................... 40 3.5 Separación y purificación ........................................................................................... 40 3.6 Pruebas de antagonismo in vitro por cultivo dual ...................................................... 41 3.6.1 Preselección de hongos endófitos ....................................................................... 41 3.7 Análisis estadísticos .................................................................................................... 42 3.8 Identificación de las morfoespecies de hongos endófitos con mayor capacidad antagónica.................................................................................................................................. 42 3.9 Evaluación de los mecanismos de antagonismo ......................................................... 42 3.10 Prueba cualitativa de Cocultivo de endófitos ............................................................. 43 4 Resultados y Discusión ........................................................................................................ 44 4.1 Aislamiento de hongos ............................................................................................... 44 4.2 Pruebas de antagonismo ............................................................................................. 47 4.3 Análisis estadístico ..................................................................................................... 51 4.3.1 Análisis de medias del crecimiento de hongos endófitos de M. oleífera en cultivo dual con P. palmarum. .......................................................................................................... 51 4.3.2 Análisis de varianza ANOVA ............................................................................. 52 4.3.3 Prueba Tukey....................................................................................................... 54 4.4 Identificación de los hongos (género) con mayor capacidad antagónica ................... 56 4.5 Evaluación de los mecanismos de antagonistas. ........................................................ 60 4.6 Prueba cualitativa de cocultivo de endófitos .............................................................. 61 5 Conclusiones ......................................................................................................................... 65 6 Bibliografía ........................................................................................................................... 67 7 ANEXOS .............................................................................................................................. 77 7.1 Anexo 1....................................................................................................................... 77 7.2 Anexo 2....................................................................................................................... 77 7.3 Anexo 3...................................................................................................................... 78 7.4 Anexo 4....................................................................................................................... 79 EVALUACIÓN IN VITRO HONGOS ENDÓFITOS 8 Lista de Tablas Pág. Tabla 1.Codificación de hongos endófitos.................................................................................... 41 Tabla 2. Aislamiento de hongos endófitos de Moringa oleífera ................................................... 45 Tabla 3.Pruebas de antagonismos de los hongos endófitos frente a Pestalotia palmarum. .......... 48 Tabla 4.Análisis de varianza de los hongos que presentaron mayor capacidad antagónica ......... 53 Tabla 5.Análisis de varianza ANOVA.......................................................................................... 54 Tabla 6.Pruebas a posteriori Tukey .............................................................................................. 55 Tabla 7.Descripción de los hongos endófitos
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