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1042770714.2019.Pdf (4.255Mb) Controladores de los biotipos de arroz y maíz de Spodoptera frugiperda (Lepidoptera, Noctuidae) en especies de Meliaceae Daniela Alejandra Álvarez Yepes Universidad Nacional de Colombia Facultad de Ciencias, Escuela de Química Medellín, Colombia 2019 Controladores de los biotipos de arroz y maíz de Spodoptera frugiperda (Lepidoptera, Noctuidae) en especies de Meliaceae Daniela Alejandra Álvarez Yepes Tesis o trabajo de investigación presentada(o) como requisito parcial para optar al título de: Magister en Ciencias Directora Tatiana Lobo Echeverri PhD. Codirector (a): Clara Inés Saldamando Benjumea PhD. Línea de Investigación: Química de los Productos Naturales Grupo de Investigación: Química de los Productos Naturales y los Alimentos, Universidad Nacional de Colombia Universidad Nacional de Colombia Facultad de Ciencias, Escuela de Química Medellín, Colombia 2019 Serendipität Agradecimientos Agradezco a la Universidad Nacional de Colombia y a la dirección de investigaciones, sede Medellín (DIME) por la financiación brindada para la ejecución de este proyecto y a Colciencias por la beca “Jóvenes Investigadores” Agradezco a mi familia y a mis amigos por todo el apoyo, amor y comprensión durante estos años. Agradezco especialmente a la profesora Tatiana Lobo por su valiosa instrucción y paciencia, a quien debo todos los instantes alegres de mi paso por el laboratorio 19A-210. Agradezco también a la profesora Clara Saldamando por su apoyo académico e investigativo. Resumen y Abstract IX Resumen Spodoptera frugiperda (Lepidoptera, Noctuidae) es una plaga polífaga de gran importancia económica en Colombia, la cual causa pérdidas hasta del 35% en los cultivos de maíz y en porcentajes menores en los cultivos de arroz, caña de azúcar, sorgo y algodón. Se ha reportado la existencia de dos biotipos de este insecto, denominados de arroz y maíz, por ser estas las plantas de las que se alimentan principalmente. Estos biotipos difieren en su susceptibilidad a insecticidas como lambadacialotrina y metomil y a las endotoxinas del Bacillus thuringensis, generalmente usados en el control de esta plaga, siendo el biotipo de arroz más resistente a los insecticidas y el biotipo de maíz a las endotoxinas. Tal diferencia en susceptibilidad supone un problema si se tiene en cuenta que ambos biotipos han adquirido resistencia a insecticidas sintéticos y a plantas transgénicas y que actualmente los pesticidas sintéticos representan un problema creciente de contaminación y riesgo a la salud. Este panorama pone de relieve la importancia de la búsqueda de alternativas biosostenibles para el control de S. frugiperda, que consideren la existencia de los biotipos. Dado el amplio espectro de compuestos con potencial insecticida e insectistático de la familia Meliaceae, se evaluó la actividad insecticida y antialimentaria de extractos vegetales de Carapa guianensis, Swietenia mahagoni y Swietenia macrophylla, usando larvas de tercer instar. De estas plantas, las fracciones de diclorometano de los tallos de C. guianensis y de S. macrophylla tuvieron mayores efectos sobre S. frugiperda, tanto insecticidas como antialimentarios, siendo S. macrophylla la más exitosa. Para evaluar un posible mecanismo de acción de la fracción se realizó un ensayo de inhibición de tirosinasa, en el que se evidenció un efecto moderado. Dicha fracción de tallos de S. macrophylla está compuesta principalmente por fitoesteroles y ácidos grasos, en la que resalta el β-sitosterol 3-O-β-glucopiranósido por ser el compuesto mayoritario y para el que se ha reportado su potencial controlador de insectos. Palabras clave: Spodoptera frugiperda, Swietenia macrophylla, Carapa guianensis, actividad insecticida, actividad antialimentaria, β-sitosterol 3-O-β-glucopiranósido. X Controladores de los biotipos de arroz y maíz de Spodoptera frugiperda (Lepidoptera, Noctuidae) en especies de Meliaceae Abstract Spodoptera frugiperda (Lepidoptera, Noctuidae) is a polyphagous pest of great economic value in Colombia, which causes losses of up to 35% in corn crops, and to a lesser extent in rice, sugarcane, cotton and sorghum crops. It has been reported that there are two strains of this insect, namely Rice and Corn strain, because these are the main plants they feed from. These strains differ from their susceptibility to insecticides like lambdacyhalothrin and methomyl, and to the Bacillus thuringensis endotoxins, generally used in this pest control, being the Rice strain more resistant to insecticides and the corn one more resistant to the endotoxins. This difference in susceptibility is a problem if it is considered that both strains have become resistant to synthetic insecticides and transgenic crops, and that nowadays the synthetic insecticides are a growing threat to human health and the environment. This scenario highlights the need to find new and bio-sustainable alternatives for controlling S. frugiperda, which consider the strains existence. Because of the wide spectrum of compounds with insecticide and insectistatic potential in the Meliaceae family, we tested the insecticide and antifeedant activity of the Carapa guianensis, Swietenia mahagoni y Swietenia macrophylla ethanolic extracts on third instar larvae. From these plants, the Carapa guianensis and Swietenia macrophylla twigs dicloromethane fractions had the major effects on S. frugiperda, both insecticide and antifeedant, being S. macrophylla the most successful. In order to evaluate a possible mechanism that explains this fraction effect we tested the S. macrophylla tyrosinase inhibition, which resulted in a moderate effect. The dicloromethane fraction is basically composed of phytosterols and fatty acids. The β-sitosterol 3-O-β-glucopyranoside is the main compound of the fraction and a potential insect controller, based on what is reported in the literature and our results. Keywords: Spodoptera frugiperda, Swietenia macrophylla, Carapa guianensis, insecticide activity, antifeedant activity, β-sitosterol 3-O-β-glucopyranoside. Contenido XI Tabla de Contenido 1. Objetivos ...................................................................................................................... 5 1.1 Objetivo general ..................................................................................................... 5 1.2 Objetivos específicos ............................................................................................. 5 2. Marco teórico y estado del arte ................................................................................. 7 2.1 Aspectos biológicos de Spodoptera frugiperda ..................................................... 7 2.1.1 Ciclo de vida de S. frugiperda ............................................................................ 8 2.1.2 Hábitos alimenticios y daño a los cultivos........................................................ 15 2.1.3 Biotipos de S. frugiperda .................................................................................. 18 2.1.4 Estrategias de control de S. frugiperda ............................................................ 25 2.2 Potencial insecticida de compuestos de origen vegetal ...................................... 34 2.3 Familia Meliaceae ................................................................................................ 42 2.3.1 Distribución geográfica y hábitat ...................................................................... 43 2.3.2 Morfología ......................................................................................................... 44 2.3.3 Importancia económica .................................................................................... 46 2.3.4 Fitoquímica ....................................................................................................... 47 2.3.5 Descripción de las especies evaluadas ........................................................... 50 3. Metodología ................................................................................................................ 63 3.1 Extracción etanólica del material vegetal ............................................................ 63 3.2 Fraccionamiento, purificación y elucidación estructural de compuestos ............ 63 3.3 Mantenimiento de insectos .................................................................................. 65 3.4 Bioensayos sobre las larvas de los biotipos de S. frugiperda ............................. 66 3.4.1 Ensayo antialimentario ..................................................................................... 66 3.4.2 Ensayo insecticida ............................................................................................ 67 3.5 Ensayo de inhibición de tirosinasa con la fracción de tallos de S. macrophylla . 67 3.6 Análisis estadístico ............................................................................................... 68 4. Resultados y discusión ............................................................................................ 70 4.1 Fraccionamiento de las especies seleccionadas ................................................ 71 4.2 Ensayo insecticida ............................................................................................... 74 4.2.1 Efectos sobre la fase larval .............................................................................. 75 4.2.2 Efectos sobre el ciclo completo ........................................................................ 94 4.3 Ensayo antialimentario ......................................................................................
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