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Tesis Doctoral, En Desarrollo UNIVERSIDAD NACIONAL DEL SUR TESIS DE DOCTORADO EN BIOLOGÍA Estrés genómico y expresión de la apomixis en pasto llorón (Eragrostis curvula (Schrad.) Nees.) Lic. Juan Manuel Rodrigo BAHÍA BLANCA ARGENTINA 2014 Prefacio PREFACIO Esta Tesis se presenta como parte de los requisitos para optar al grado Académico de Doctor en Biología, de la Universidad Nacional del Sur y no ha sido presentada previamente para la obtención de otro título en esta Universidad u otra. La misma contiene los resultados obtenidos en investigaciones llevadas a cabo en el ámbito del Departamento de Biología, Bioquímica y Farmacia durante el período comprendido entre octubre de 2009 y junio de 2014, bajo la dirección de la Dra. Viviana Echenique y la dirección adjunta del Dr. Diego Zappacosta. Lic. Juan Manuel Rodrigo 3 Agradecimientos Agradecimientos Al Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) por haberme otorgado el sustento económico para realizar mis estudios de doctorado. Asimismo deseo agradecer a las autoridades del CERZOS y el Dpto. de Biología, Bioquímica y Farmacia de la Universidad Nacional del Sur por haberme brindado el espacio de trabajo. A Viviana, mi directora de tesis, por estar siempre dispuesta a enseñarme, discutir y brindarme sus cocimientos y experiencia en mi formación como investigador, y sobre todo gracias por confiar en mí. A mi co-director, Diego, por sus valiosas sugerencias, todos los conocimientos brindados y por su invaluable ayuda en el laboratorio. A todos los integrantes del Laboratorio de Biotecnología y Genética Vegetal, por hacer del lugar de trabajo un lugar en el que uno realmente desea estar, gracias por los gratos momentos compartidos. A Manfred, Dani, Diego y Juan, por su amistad y la ayuda que me dieron durante la realización de este trabajo. A Luis Hernández e Ivone Lindström, por haberme dado la posibilidad de trabajar en su laboratorio, gracias por haber sido tan atentos, tanto en la relación laboral como en lo personal. A mis padres, Nilda y José Luis, por la formación que me dieron, por alentarme siempre en el camino del estudio y el trabajo, y por estar siempre a mi lado. A mis hermanos, Lucila, Laureano y Candela, por todo el amor que me brindan y por ser las maravillosas personas que son. Gracias por estar. A Carmen y Juan Carlos, por todo su cariño y el alimento brindado. A mis amigos, siempre dispuestos a darme una mano en cualquier momento si lo necesitaba. 4 Agradecimientos A Lorena, mi mujer, por su contención, su comprensión y su paciencia durante el desarrollo de esta tesis. Gracias mi amor por todo el amor que me das, por estar conmigo siempre y especialmente gracias por darle sentido a mi vida. A Bruno y Lana, por darme amor. A todos ustedes, sinceramente muchas gracias. Juan. 5 Publicaciones generadas durante el desarrollo de la tesis Publicaciones generadas durante el desarrollo de la tesis Publicaciones en revistas científicas Zappacosta D., Ochogavía A., Rodrigo J.M., Romero J., Meier M., Polci P., Garbus I., Pessino S., Echenique V. (2014) Increased apomixis expression concurrent with genetic and epigenetic variation in a newly synthesized Eragrostis curvula polyploid. Scientific reports, 4. Publicaciones en Revistas de Divulgación: Echenique V., Pessino S.C., Meier M., Selva J.P., Ochogavia A., Zappacosta D., Romero J.R., Rodrigo J.M., Garbus I., Terenti C., Diaz A. (2012) Apomixis en Eragrostis curvula. Ciencias Agronómicas, 20: 77. Rodrigo J.M., Zappacosta D., Bonaventure G., Echenique V. (2012) Efectos del estrés abiótico sobre la expresión de la apomixis en pasto llorón. Ciencias Agronómicas, 20: 79. Zappacosta D., Meier M., Terenti C., Rodrigo J.M., Echenique V. (2012) Poblaciones de mapeo en pasto llorón - éxitos y fracasos. Ciencias Agronómicas 20: 81. Rodrigo J.M., Selva J.P., Zappacosta D., Echenique V. (2013) Epigenética, revolucionando la expresión génica. AgroUNS, (En Prensa). Presentaciones a congresos Internacionales Rodrigo J.M., Ochogavia A., Meier M., Zappacosta D., Pessino S., Echenique V. “Effects of stress and genomic plasticity on the reproductive mode of facultative apomictic plants of weeping lovegrass [Eragrostis curvula (Schrad.) Nees]”. The 6th international symposium on the molecular breeding of forage and turf. 15-19 de marzo 2010, Buenos Aires, Argentina. Rodrigo J.M., Orellano N, Selva JP, Echenique V, Zappacosta D. “Determinación del modo reproductivo de Tetrachne dregei Ness.”. XV Congreso Latinoamericano de Genética. 28 - 31 de octubre de 2012. Rosario, Santa Fe, Argentina. 6 Publicaciones generadas durante el desarrollo de la tesis Meier M., Rodrigo J.M., Zappacosta D., Echenique V. “Efecto mentor en pasto llorón, Eragrostis curvula”. XV Congreso Latinoamericano de Genética. 28 - 31 de octubre de 2012. Rosario, Santa Fe, Argentina. Rodrigo J.M., Zappacosta D., Echenique V. “Evaluación de la estabilidad reproductiva en pasto llorón (Eragrostis curvula Nees).”. XV Congreso Latinoamericano de Genética. 28 - 31 de octubre de 2012. Rosario, Santa Fe, Argentina. Nacionales Rodrigo J.M., Marinangeli P.A., Curvetto N.R. “Propagación vegetativa de Rhodophiala bifida”. XI Jornadas Nacionales de Floricultura. 7-9 de octubre 2009. Montecarlo, Misiones. SELECCIONADO PARA PRESENTACIÓN ORAL Rodrigo J.M., Meier M., Zappacosta D., Echenique V. “Expresión de la apomixis bajo estrés hídrico en pasto llorón”. XL Congreso Argentino de Genética y I Jornadas Regionales SAG-NEA. 18-21 de septiembre 2011. Corrientes, Corrientes. 7 Índice Índice PREFACIO ................................................................................................................................................. 3 AGRADECIMIENTOS ........................................................................................................................... 5 PUBLICACIONES GENERADAS DURANTE EL DESARROLLO DE LA TESIS ……………. 6 RESUMEN ............................................................................................................................................... 11 ABSTRACT ............................................................................................................................................. 14 INTRODUCCIÓN ……………............................................................................................................. 17 Las plantas y el estrés .................................................................................................................. 18 Del estrés ambiental al estrés genómico ……………............................................................... 19 Efecto del estrés sobre el modo reproductivo ......................................................................... 21 La apomixis en plantas ………………………………………………...……………………… 23 Mecanismos de apomixis ……………………………………………………………………… 27 Control genético de la apomixis ……………………………………………………………… 30 Apomixis y Poliploidía ………………………………………………………………………. 31 Filogenia y distribución geográfica de la apomixis ………………………………………… 33 Apomixis en pasto llorón …………………………………………………………………….. 34 Importancia de la Apomixis en la Agricultura …………………………………………….. 38 Estrategias para el desarrollo de cereales apomícticos ……………………………………. 40 Eragrostis curvula (Pasto llorón, weeping lovegrass) ….......................................................... 41 Apomixis y estrés ……………………………............................................................................. 45 HIPÓTESIS ............................................................................................................................................. 49 OBJETIVOS ............................................................................................................................................ 51 Objetivo general ......................................................................................................................... 52 Objetivos particulares ................................................................................................................ 52 CAPÍTULO I - “Expresión de la apomixis en pasto llorón durante el periodo de floración.”.... 53 Introducción ................................................................................................................................ 54 Hipótesis ...................................................................................................................................... 62 Objetivo ....................................................................................................................................... 62 Materiales y Métodos ................................................................................................................ 63 Material Vegetal .......................................................................................................... 63 Estudios citoembriológicos ....................................................................................... 63 Análisis estadísticos .................................................................................................... 64 Resultados y Discusión ............................................................................................................. 64 Estudios citoembriológicos ....................................................................................... 64 Análisis del desarrollo de la inflorescencia ……………………………………… 69 8 Índice CAPÍTULO II - “Expresión de la apomixis bajo diferentes situaciones que provocan estrés genómico.”................................................................................................................................................
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