Resumen de Genetic and genomic approaches to characterize crop varieties
RESUMEN DE LA TESIS DOCTORAL DE D./Dª Leticia Ayllón Egea El resumen de la tesis para la base de datos Teseo debe ser una presentación de la tesis y tener la extensión suficiente para que quede explicado el argumento de la tesis doctoral. El formato debe facilitar la lectura y comprensión del texto a los usuarios que accedan a Teseo, debiendo diferenciarse las siguientes partes de la tesis:
1. Introducción o motivación de la tesis Los bancos de germoplasma de plantas son reservorios de biodiversidad. Pueden albergar variedades en múltiples formas, como semillas, arboretos, sembrando cultivos estacionales y manteniendo cultivos in vitro. Los registros de los primeros bancos de germoplasma pertenecen a las sociedades egipcia y babilónica. Probablemente, Nikolaj Ivanovič Vavilov fue la primera persona que recalcó la necesidad de crear bancos de germoplasma para el bienestar de la sociedad. Tradicionalmente, las variedades eran almacenadas en función de las características morfológicas. Sin embargo, debido a la plasticidad fenotípica de las plantas, esta clasificación puede dar lugar a sinonimias y homonimias. Esta situación ha desencadenado la necesidad de caracterizar los bancos de germoplasma, no solo por criterios morfológicos, sino también mediante marcadores moleculares. Recientemente, estos últimos han mejorado significativamente. De este modo, han evolucionado desde métodos basados en péptidos a otros basados en ADN. Es más, estos últimos han mejorado enormemente gracias a la reacción en cadena de la polimerasa (“PCR”), incluyendo aproximaciones de alto rendimiento recientes. Esto ha sido posible gracias a la emergencia de tecnologías como las plataformas de secuenciación de segunda generación (“SGS”) y de tercera generación (“TGS”). Las aproximaciones utilizadas en la actualidad incluyen las repeticiones de secuencias únicas (“SSR”), los polimorfismos de nucleótidos únicos (“SNP”) y el genotipado por secuenciación (“GBS”). En resumen, la caracterización molecular permite una mejor identificación genética, comprensión de las funciones biológicas y búsqueda de relaciones para biología evolutiva. También son utilizadas en biología de la conservación de plantas, estudios de bioseguridad y gestión de bancos de germoplasma. Además, son herramientas excelentes para la mejora asistida, así como para la certificación de la propiedad intelectual y aplicaciones en trazabilidad.
2.Contenido de la investigación En esta Tesis Doctoral, bancos de germoplasma de ajo y olivo han sido analizados mediante marcadores moleculares. Brevemente, el ajo ha sido tradicionalmente utilizado en todo el mundo como un ingrediente común en alimentación y como un remedio natural en farmacología y medicina. Esto es debido a sus interesantes atributos beneficiosos como, la reducción de la tensión alta, del colesterol y en la arterioesclerosis. El ajo también tiene efectos preventivos contra el cáncer y actividad antimicrobiana. Por otro lado, la importancia del olivo en innegable. Es uno de las especies más cultivadas en el mundo, especialmente en la cuenca mediterránea, y es la segunda especie más utilizada en la producción de aceite tras el aceite de palma. El área total de cultivo es diez millones de hectáreas. Sus innumerables propiedades culinarias y medicinales han impulsado su expansión a áreas que no tienen tradición productora ni consumidora. El principal objetivo de la caracterización del banco de germoplasma de ajo era estudiar la diversidad y estructura genética de 417 muestras del principal bando de germoplasma de ajo localizado en el Instituto Andaluz de Investigación y Formación Agraria, Pesquera, Alimentaria y de la Producción Ecológica (IFAPA) de la Junta de Andalucía, de la Universidad de Córdoba y del Centro de Ensayos de Evaluación de Variedades, localizado en el Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) en Madrid. La técnica elegida fue DArTseq (tecnología de matriz de diversidad), la cual permite la caracterización en especies sin genoma de referencia u otra información genética previa. De este modo, una colección nuclear fue creada para reducir el número original de entradas del banco, sin perder diversidad genética. Además, las secuencias polimórficas de ajo generadas en el análisis de DArTseq fueron usadas para determinar sus identidades, funciones, términos de Ontología Génica (“GO”) y rutas metabólicas mediante la búsqueda de identidades en otras bases de datos de plantas. Por otro lado, el objetivo principal del capítulo de olivo fue describir un método de “tubo cerrado” rentable para genotipar variedades cuando existe información genética disponible. En este caso, 83 muestras fueron analizadas usado seis marcadores moleculares y el análisis de alta resolución de fusión (“HRM”). Además, aunque el número de marcadores empleado fue bajo, los análisis de caracterización estaban en concordancia con trabajos previos.
3.Conclusión Ambas técnicas de marcadores moleculares (DArTseq y HRM) mostraron resultados de genotipado consistentes en función de la información previa de los pasaportes de datos. El tamaño del banco de germoplasma de ajo fue significativamente reducido, lo que indica que en análisis DArTseq es una tecnología adecuada para el genotipado de alto rendimiento sin información genética previa disponible. Hasta nuestro conocimiento, este es el primer genotipado por secuenciación de alto rendimiento en ajo mediante la tecnología DArTseq. El genotipado mediante análisis de HRM en olivo demostró ser una metodología rentable para la caracterización de germoplasma y los estudios de genotipado. Los análisis realizados en estos capítulos pueden ayudar a clarificar los estudios genéticos y las aproximaciones para estudiar las adaptaciones para hacer frente a estreses bióticos y abióticos. Lo que es particularmente relevante en el contexto actual de cambio climático y calentamiento global.
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