Determinación de la variabilidad genética de 50 clones de cacao de la colección allen del INIAP con la utilización de marcadores moleculares microsatélites
Fecha
2014
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Facultad de Ciencias Agrarias Universidad de Guayaquil
Resumen
Existen diversas herramientas para la caracterización ya sea del tipo morfológico como molecular. Para esta investigación, que tiene como base la caracterización molecular, se escogieron los marcadores SSR basándose en los objetivos planteados. Por su característica de codominantes, los marcadores microsatélites brindan una herramienta versátil para los fitomejoradores al momento de realizar segregaciones de descendencia. Se tiene como objetivos de este estudio la caracterización molecular de 50 individuos de cacao de la colección Allen. Además, determinar variantes como el poder discriminante de los marcadores SSR, estos basados en la técnica de PCR. La colecta de los clones se la realizó en la EETP (Estación Experimental Tropical Pichilingue) del INIAP, ubicada en la provincia de Los Ríos, donde los árboles de esta colección se encuentran en tres lotes distintos. Se colectaron hojas fotosintéticas en buenas condiciones; mediante pruebas preliminares se determinó el protocolo de extracción llevando a cabo el descrito por Doyle y Doyle. Luego de la obtención y pruebas de los resultantes de la extracción se procedió a la amplificación de las muestras, las mismas que fueron amplificadas con el ciclaje establecido por la Estación Experimental Santa 53 Catalina para marcadores microsatélites, con variación en la temperatura de anillamiento. Se realizó el test de integridad para las muestras, obteniendo buena amplificación de las imágenes. La secuenciación de las muestras se la efectúo en el LICOR 4300. Se realizó la documentación de los geles para luego realizar el genotipage en el programa SAGA; se transformó a matrices de datos, estos resultados fueron modificados y depurados, basándose en trabajos antes realizados donde se le podía asignar un valor ya conocido a los alelos encontrados. Se convirtió en una matriz binaria donde se pudo establecer la relación de
similitud entre los individuos, con la utilización del programa Power Markert. Utilizando los postulados descritos por Nei (1973), se logró establecer la diversidad genética dentro de la colección. Para la elaboración de los árboles de agrupamiento entre los clones se usó el coeficiente de UPGMA que utiliza el ligamento promedio entre individuos, proyectando resultados satisfactorios para futuros estudios de mejoramiento.
Several tools for characterizing either the morphological and molecular type, for this research is based molecular characterization SSR markers were chosen based on the objectives. For his feature codominant microsatellites markers provide a versatile tool for breeders at the time of descent Fito segregations are aims of this study, the molecular characterization of 50 individuals of cocoa Allen collection. Also variants such as determining the discriminating power of these SSR markers based on PCR, the collection of the collection was made in the EETP (Pichilingue Tropical Station) Los Rios province where the trees in this collection are in three different lots. Photosynthetic leaves were collected in good condition mediantes pretesting the extraction protocol was determined by carrying just described by Doyle and Doyle, after obtaining and testing resulting from the extraction, we proceeded to the amplification of the samples the same as were amplified with the cycling set by Santa Catalina Experimental Station for microsatellite markers with variation in the annealing temperature, the integrity test samples to obtain good amplification in the images presented were performed. The sequencing of the samples was the conducted at the LIQUOR 4300 documentation gels then perform genotipage in the SAGA program was conducted, it became a data matrix, these results were modified and depurándos based on work formerly done where he you could assign a value already known alleles found became a binary matrix where it could 55 establish the relationship of similarity between individuals with the use of Power Markert program. Using the principles described by Nei was able to establish the genetic diversity within the collection, for the production of trees of groups of clones coefficient UPGMA using the average ligament between individuals was used, projecting satisfactory results for future studies of improvement .
Several tools for characterizing either the morphological and molecular type, for this research is based molecular characterization SSR markers were chosen based on the objectives. For his feature codominant microsatellites markers provide a versatile tool for breeders at the time of descent Fito segregations are aims of this study, the molecular characterization of 50 individuals of cocoa Allen collection. Also variants such as determining the discriminating power of these SSR markers based on PCR, the collection of the collection was made in the EETP (Pichilingue Tropical Station) Los Rios province where the trees in this collection are in three different lots. Photosynthetic leaves were collected in good condition mediantes pretesting the extraction protocol was determined by carrying just described by Doyle and Doyle, after obtaining and testing resulting from the extraction, we proceeded to the amplification of the samples the same as were amplified with the cycling set by Santa Catalina Experimental Station for microsatellite markers with variation in the annealing temperature, the integrity test samples to obtain good amplification in the images presented were performed. The sequencing of the samples was the conducted at the LIQUOR 4300 documentation gels then perform genotipage in the SAGA program was conducted, it became a data matrix, these results were modified and depurándos based on work formerly done where he you could assign a value already known alleles found became a binary matrix where it could 55 establish the relationship of similarity between individuals with the use of Power Markert program. Using the principles described by Nei was able to establish the genetic diversity within the collection, for the production of trees of groups of clones coefficient UPGMA using the average ligament between individuals was used, projecting satisfactory results for future studies of improvement .
Descripción
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Palabras clave
CACAO, CLONES, VARIABILIDAD GENETICA
Citación
CHenche López, Óscar Mauricio (2014): Determinación de la variabilidad genética de 50 clones de cacao de la colección allen del INIAP con la utilización de marcadores moleculares microsatélites