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Aspectos Reproductivos, Citogenéticos Y Evolutivos Aplicados Al Mejoramiento En El Género Passiflora L

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Aspectos Reproductivos, Citogenéticos Y Evolutivos Aplicados Al Mejoramiento En El Género Passiflora L Tesis Doctoral Aspectos reproductivos, citogenéticos y evolutivos aplicados al mejoramiento en el género Passiflora L. Bugallo, Verónica Lucía 2017 Este documento forma parte de las colecciones digitales de la Biblioteca Central Dr. Luis Federico Leloir, disponible en bibliotecadigital.exactas.uba.ar. Su utilización debe ser acompañada por la cita bibliográfica con reconocimiento de la fuente. This document is part of the digital collection of the Central Library Dr. Luis Federico Leloir, available in bibliotecadigital.exactas.uba.ar. It should be used accompanied by the corresponding citation acknowledging the source. Cita tipo APA: Bugallo, Verónica Lucía. (2017). Aspectos reproductivos, citogenéticos y evolutivos aplicados al mejoramiento en el género Passiflora L.. Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. https://hdl.handle.net/20.500.12110/tesis_n6250_Bugallo Cita tipo Chicago: Bugallo, Verónica Lucía. "Aspectos reproductivos, citogenéticos y evolutivos aplicados al mejoramiento en el género Passiflora L.". Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. 2017. https://hdl.handle.net/20.500.12110/tesis_n6250_Bugallo Dirección: Biblioteca Central Dr. Luis F. Leloir, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires. Contacto: bibliotecadigital.exactas.uba.ar Intendente Güiraldes 2160 - C1428EGA - Tel. (++54 +11) 4789-9293 UNIVERSIDAD DE BUENOS AIRES Facultad de Ciencias Exactas y Naturales Departamento de Ecología, Genética y Evolución Aspectos reproductivos, citogenéticos y evolutivos aplicados al mejoramiento en el género Passiflora L. Tesis para optar al título de Doctora de la Universidad de Buenos Aires en el área Ciencias Biológicas Ing. Agr. Verónica Lucía Bugallo Directoras de Tesis: Dra. Lidia Poggio & Dra. Gabriela Facciuto Consejera de Estudios: Dra. María Isabel Remis Laboratorio de Citogenética y Evolución - Departamento de Ecología, Genética y Evolución - Facultad de Ciencias Exactas y Naturales - Universidad de Buenos Aires. Instituto de Floricultura - INTA. Cátedra de Genética - Facultad de Agronomía - Universidad de Buenos Aires. Fecha de defensa: 2 de agosto de 2017 Ciudad Autónoma de Buenos Aires I Agradecimientos A las instituciones que me respaldaron: Facultad de Agronomía, U.B.A.; Laboratorio de Citogenética y Evolución, Facultad de Ciencias Exactas y Naturales, U.B.A. e Instituto de Floricultura del I.N.T.A. A mis directoras. Al jurado: Sara Maldonado, Gabriel Rua y Sandra Aliscioni. A los amigos que colaboraron en esta tesis: Florencia Realini, Marisa Bonasora, Hernán Burrieza, María Julia Pannunzio, Carolina Castagnet y Ramiro Oviedo. II Índice Introducción general………………………………………………………………….…..1 Capítulo 1: Las especies argentinas del género Passiflora……………………….9 Introducción………………………………………………………………………...9 Materiales y métodos…………………………………………………………...15 Resultados…………………………………………………………………………25 Discusión…………………………………………………………………………..69 Conclusiones………………………………………………………………………81 Capítulo 2: Hibridación interespecífica entre especies nativas del género Passiflora subgénero Passiflora…………………………………….......................83 Introducción……………………………………………………………………….83 Materiales y métodos…………………………………………………………….87 Resultados…………………………………………………………………………90 Discusión…………………………………………………………………………100 Conclusiones…………………………………………………………………….109 Capítulo 3: Estudios citogenéticos en híbridos interespecíficos de Passiflora…………………………………………………………………………………111 Introducción……………………………………………………………………..111 I Materiales y métodos…………………………………………………………115 Resultados………………………………………………………………………118 Discusión………………………………………………………………………..162 Conclusiones……………………………………………………………………175 Capítulo 4: Estudio de caracteres fenotípicos de importancia ornamental en especies e híbridos de Passiflora…………………………..............................177 Introducción…………………………………………………………………….177 Materiales y métodos…………………………………………………………181 Resultados………………………………………………………………………185 Discusión………………………………………………………………………..228 Conclusiones……………………………………………………………………236 Capítulo 5: Inducción de poliploides en Passiflora…………………………….238 Introducción…………………………………………………………………….238 Materiales y métodos………………………………………………………….243 Resultados……………………………………………………………………….249 Discusión…………………………………………………………………………266 Conclusiones…………………………………………………………………….271 Conclusiones generales……………………………………………………………….273 II Bibliografía……………………………………………………………………………….277 Anexo 1: Modelado del nicho ecológico en especies nativas de Passiflora…300 Anexo 2: Soluciones y buffers empleados para estudios citológicos………..368 Anexo 3: Análisis estadísticos sobre la cantidad de ADN en especies de Passiflora…………………………………………………………………………………371 Anexo 4: Análisis estadísticos sobre hibridación interespecífica en Passiflora…………………………………………………………………………………374 Anexo 5: Híbridos entre especies argentinas de Passiflora publicados hasta la fecha……………………………………………………………………………………….376 Anexo 6: Análisis estadísticos sobre citogenética de híbridos interespecíficos………………………………………………………………………….379 Anexo 7: Análisis estadísticos sobre cantidad de ADN en híbridos interespecíficos……………………………………………………………………….…381 Anexo 8: Análisis estadísticos sobre caracteres fenotípicos…………………..383 Anexo 9: Registros de la evaluación de tolerancia a temperaturas invernales de Buenos Aires………………………………………………………………………...404 Anexo 10: Formulario y cuestiorario técnico para el registro de la variedad Passiflora “Amatista” INTA…………………………………………………………...406 Anexo 11: Análisis estadíticos sobre inducción de poliploides en Passiflora…………………………………………………………………………………434 III Resumen El género Passiflora cuenta con más de 500 especies, de la cuales 19 son nativas de Argentina. Este género está incluido en un programa de mejoramiento que tiene como objetivo la obtención de variedades ornamentales con características fenotípicas novedosas y con tolerancia a bajas temperaturas. En el análisis citogenético de catorce especies nativas, se reportó por primera vez el número somático 2n = 18 para P. tenuifila y para un citotipo de P. mooreana. Además, se confirmaron los números básicos x = 9 para el subgénero Passiflora, x = 6 para Decaloba y x = 10 para Dysosmia. Los parámetros cariotípicos obtenidos mediante técnicas clásicas y moleculares (FISH) permitieron elaborar los idiogramas de todas las especies y establecer diferencias entre los subgéneros. La hibridación in situ genómica (GISH) reveló que el subgénero Passiflora posee mayor afinidad genómica con el subgénero Dysosmia que con Decaloba. La cantidad de ADN estimada mediante citometría de flujo segregó a los subgéneros con menores cantidades de ADN (0,5 a 0,72 pg.), Dysosmia y Decaloba, respecto de Passiflora (0,9 a 2,5 pg.). Se encontraron barreras de aislamiento reproductivo que actuarían en condiciones naturales (diferencias de nicho ecológico y en los períodos de floración) y otras que actuarían dificultando la formación de híbridos en el programa de mejoramiento (incompatibilidad unilateral, ausencia de formación de frutos, fallas en la germinación de las semillas e inviabilidad de plántulas). IV La especie P. amethystina fue muy prolífica, produciendo frutos con seis de los ocho taxones con los que se la cruzó. Por otro lado, P. alata fue la especie con la que se registró la mayor cantidad de cruzamientos inviables. En los estudios meióticos, los híbridos que presentaron los porcentajes máximo y mínimo de meiocitos con 9 bivalentes fueron P. amethystina x P. edulis f. flavicarpa y P. amethystina x P. alata (95 y 40%, respectivamente). Algunos híbridos estériles presentaron meiosis regulares y elevados índices meióticos, fenómeno que se explicaría por la hipótesis de la hibridez estructural críptica. Además, híbridos altamente estériles presentaron meiosis irregular, puente y fragmento en anafase I, husos tripolares y citomixis. La mayor viabilidad del polen en híbridos se registró en P. amethystina x P. caerulea (90,4%) y P. amethystina x P. edulis f. edulis (79,4%). La técnica de hibridación in situ (GISH), permitió reconocer los genomas parentales en los híbridos P. alata x P. caerulea, P. alata x P. cincinnata, P. alata x P. amethystina y P. mooreana x P. cincinnata, por lo que se infiere que existen diferencias en ADN repetido entre las especies progenitoras de cada híbrido. El contenido de ADN de los híbridos no siempre coincidió con el promedio del tamaño genómico de sus parentales. Esto podría deberse a cambios revolucionarios tales como eliminación/ganancia de secuencias repetidas codificantes y no codificantes. Estudios fenotípicos mostraron que la superficie de las hojas y el tamaño de las flores en híbridos fueron intermedios a los de los parentales en V la mayoría de los cruzamientos estudiados, mostrando herencia cuantitativa, mientras que en otros se registró una herencia transgresiva de estos caracteres, sugiriendo heterosis. Además, se logró la inducción de poliploides a partir del tratamiento con Colchicina de estacas enraizadas y plántulas de especies e híbridos. Las pruebas de tolerancia a las bajas temperaturas y la estimación del potencial de la cobertura de estructuras para cortinas y paredes verdes permitieron seleccionar especies e híbridos, uno de los cuales se encuentra en proceso de registro como la primera variedad del género en Argentina. Palabras clave: Passiflora, plantas nativas de Argentina, hibridación interespecífica, citogenética, mejoramiento de plantas ornamentales, inducción de poliploides. VI Reproductive, cytogenetic and evolutionary aspects applied to breeding in the
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