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Centro De Ciencias Agropecuarias Departamento De Biología CENTRO DE CIENCIAS AGROPECUARIAS DEPARTAMENTO DE BIOLOGÍA Tesis FILOGENIA DEL COMPLEJO BOUTELOUA REPENS (POACEAE: CHLORIDOIDEAE) BASADA EN DATOS MOLECULARES Presenta BIOL. GERALDINE MURILLO PÉREZ Para obtener el grado de MAESTRA EN CIENCIAS AGRONÓMICAS TUTOR María Elena Siqueiros Delgado PhD. COMITÉ TUTORAL J. Travis Columbus PhD. Dr. Gilberto A. Ocampo Acosta PhD. Aguascalientes, Ags. Mayo de 2017 AGRADECIMIENTOS A la Universidad Autónoma de Aguascalientes por brindarme las herramientas necesarias para desempeñar mi profesión con responsabilidad y sentido humano, y al Consejo Nacional de Ciencia y Tecnología por la beca número 614403, otorgada para el desarrollo de mis estudios de posgrado. A mi tutora, la Dra. María Elena Siqueiros, por su apoyo incondicional para el desarrollo de todas mis metas profesionales y personales; por siempre agotar todos los recursos a su alcance para realizar y concluir satisfactoriamente mi proyecto de investigación; y sobre todo, por ser el pilar más importante en mi formación profesional y por su invaluable amistad. A mis asesores, el Dr. Travis Columbus y el Dr. Gilberto Ocampo, por el vasto conocimiento que han compartido conmigo desde la concepción de éste proyecto, por sus consejos, sugerencias y por la disposición mostrada para llevar a cabo exitosamente éste proyecto. Al personal del laboratorio de Biotecnología vegetal de la UAA, por facilitarme el uso de sus instalaciones y por apoyarme en todo lo que necesité. A los trabajadores, investigadores y estudiantes del Rancho Santa Ana Botanic Garden, por acogerme como un miembro más de su equipo, por los consejos que me brindaron dentro del laboratorio de biología molecular, por compartirme su conocimiento y experiencia en el campo de la sistemática y filogenia vegetal; y sobre todo, por hacerme sentir como en casa. A los encargados e investigadores de los herbarios IEB y MEXU, por su disposición y apoyo para la toma de muestras de los ejemplares depositados en sus colecciones, que se incluyeron en el presente trabajo. A mis compañeros y amigos del herbario HUAA, por estar siempre dispuestos a escuchar mis inquietudes y dudas, y ayudarme dentro de sus posibilidades a resolverlas y enriquecer mi proyecto con sus recomendaciones y observaciones. Al personal de los herbarios IEB y MEXU por permitirme la extracción de material vegetal para realizar los estudios filogenéticos incluidos en el presente trabajo. A mis compañeros de colecta Julio, Fer, Marce, Marisol, Beto y los doctores Columbus y Siqueiros quienes hicieron esos largos recorridos más amenos y divertidos. A las biólogas Marisol Pacheco Jiménez y María Elena Mendoza López por su valiosa colaboración en el trabajo de laboratorio realizado durante el proyecto, por su amistad y por su apoyo incondicional en el ámbito personal y profesional. Y al Ing. Carlos Eduardo Hernández Rincón, por proporcionar el equipo computacional necesario para la ejecución de todos los análisis filogenéticos incluidos en el presente trabajo. DEDICATORIAS A mis padres, quienes a pesar de la distancia siempre han sabido estar presentes en todos mis momentos de triunfo y derrota, ayudándome a levantarme cuando lo he necesitado y a celebrar las metas conquistadas; a quienes les debo no solo la vida, sino ser la mujer que soy, alguien que no se rinde. A mis hermanas, mi mayor fuente de alegrías y orgullo, y mi más grande inspiración en la vida. A mi tío Chuy, mi confidente, mi consejero, mi ángel guardián, quien siempre ha estado a mi lado cuidándome y guiándome por el mejor camino. A mi mejor amigo, compañero y cómplice, Carlos, que aún a la distancia siempre ha estado a mi lado apoyándome sin condiciones y motivándome a cumplir todas mis metas; escuchándome y brindándome su paciencia y consejos, consolándome en los momentos más difíciles y levantándome cuando he caído. A ti, por llegar a mi vida cuando más te necesitaba, y por siempre demostrarme tu cariño y amor sinceros. ÍNDICE GENERAL ÍNDICE GENERAL........................................................................................................... 1 ÍNDICE DE TABLAS ....................................................................................................... 3 ÍNDICE DE FIGURAS ...................................................................................................... 4 TABLA DE ANEXOS ....................................................................................................... 5 RESUMEN ......................................................................................................................... 6 ABSTRACT ....................................................................................................................... 7 INTRODUCCIÓN ............................................................................................................. 8 ANTECEDENTES ........................................................................................................... 11 MARCO TEÓRICO ......................................................................................................... 13 Uso de caracteres moleculares ....................................................................................... 13 Espaciador transcrito interno (ITS) del ADN ribosomal nuclear (nrADN). ................. 14 Espaciador intergénico trnT-L-F ................................................................................... 16 PLANTEAMIENTO DEL PROBLEMA ........................................................................ 18 OBJETIVOS .................................................................................................................... 19 Objetivo general ............................................................................................................. 19 Objetivos específicos ..................................................................................................... 19 HIPÓTESIS DE INVESTIGACIÓN ............................................................................... 19 CAPÍTULO I. FILOGENIA MOLECULAR .............................................................. 20 MATERIALES Y MÉTODOS ........................................................................................ 20 Muestras analizadas ................................................................................................ 20 Extracción de ADN ................................................................................................ 25 Marcadores moleculares utilizados ........................................................................ 26 Gen nuclear ................................................................................................. 26 Gen de cloroplasto ...................................................................................... 26 Condiciones de amplificación del ADN ................................................................. 27 Gen nuclear ................................................................................................. 27 Gen de cloroplasto ...................................................................................... 28 Secuenciación de ADN ........................................................................................... 28 Caracterización de las secuencias ........................................................................... 29 1 Análisis filogenético ............................................................................................... 30 RESULTADOS ................................................................................................................ 31 DISCUSION .................................................................................................................... 37 CAPÍTULO II. EVOLUCIÓN DE CARACTERES MORFOLÓGICOS ................ 40 MATERIALES Y MÉTODOS ........................................................................................ 43 RESULTADOS ................................................................................................................ 45 Presencia/Ausencia de rizoma. ............................................................................... 45 Forma de la flor rudimentaria ................................................................................. 45 Número de espiguillas por rama ............................................................................. 50 Longitud de la arista media de la lema de la flor rudimentaria .............................. 50 DISCUSION .................................................................................................................... 53 Presencia/Ausencia de Rizoma ............................................................................... 53 Forma de la flor rudimentaria ................................................................................. 53 Número de ramas por inflorescencia ...................................................................... 54 Número de espiguillas por rama ............................................................................. 55 Longitud de la arista media del rudimento ............................................................. 56 CAPITULO III. BIOGOEGRAFÍA HISTÓRICA ..................................................... 60 MATERIALES Y MÉTODOS ........................................................................................ 63 RESULTADOS ................................................................................................................ 64 DISCUSION ...................................................................................................................
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