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Centro De Investigación Científica De Yucatán, AC Centro de Investigación Científica de Yucatán, A.C. Posgrado en Ciencias Biológicas Evidencia de la transmisión del Papaya meleira virus (PMeV) por insectos en Carica papaya L. Tesis que presenta GUADALUPE ISABEL GARCÍA CÁMARA En opción al título de DOCTOR EN CIENCIAS BIOLÓGICAS (Opción Bioquímica y Biología molecular) Mérida, Yucatán, México Octubre del 2018 AGRADECIMIENTOS A la Dra. Daisy de la Caridad Pérez Brito, por darme la oportunidad de formar parte de su equipo al inicio de este proyecto, por su dedicación y sus valiosas enseñanzas durante mi formación académica. Al Dr. Óscar Alberto Moreno Valenzuela, por su apoyo, su confianza y su valiosa dirección a lo largo de todo mi proceso académico. Al Dr. Raúl Tapia Tussell, por sus comentarios y enseñanzas, las cuales contribuyeron al mejoramiento de mi desarrollo académico. Al Dr. Anuar Ahmed Magaña Álvarez, por su apoyo durante cada etapa de la presente investigación y su valiosa amistad. Al I.Q.I. Alberto Cortés Velázquez, por su orientación sobre las técnicas moleculares durante la fase experimental de la presente tesis de doctorado. Al M.C. Rodolfo Martín Mex, por su asesoría sobre el manejo agronómico de las especies vegetales durante los experimentos de campo del presente trabajo de investigación. Al Consejo Nacional de Ciencia y Tecnología (CONACYT) por el otorgamiento de la beca de manutención número 265330, sin la cual no hubiera sido posible realizar mis estudios de doctorado. Al Centro de Investigación Científica de Yucatán A.C., por permitirme el uso de sus instalaciones a lo largo de toda mi formación académica. Al posgrado en Ciencias Biológicas y la Coordinación de la opción en Bioquímica y Biología Molecular, por su apoyo y orientación durante los procesos de titulación. Al Laboratorio GeMBio (Grupo de Estudios Moleculares aplicados a la Biología), por permitirme el uso de sus instalaciones y equipos, y proporcionarme los materiales necesarios para el desarrollo de la presente investigación. A mi Comité Tutoral: Dra. Daisy de la Caridad Pérez Brito, Dr. Óscar Alberto Moreno Valenzuela, Dra. Patricia Machado Bueno Fernandes y Dr. Raúl Tapia Tussell, por su dedicación y sus valiosas observaciones, las cuales contribuyeron al fortalecimiento de mi formación académica. A mi comité predoctoral: Dra. Patricia Machado Bueno Fernandes, Dr. Raúl Tapia Tussell, Dra. Cecilia Hernández Zepeda, Dr. Jairo Cristóbal Alejo y Dr. Enrique Castaño de la Serna, por sus importantes comentarios dirigidos al mejoramiento de mi desarrollo académico. A mi Comité Revisor: Dra. Daisy de la Caridad Pérez Brito, Dr. Óscar Alberto Moreno Valenzuela, Dra. Patricia Machado Bueno Fernandes, Dr. Raúl Tapia Tussell, Dra. Cecilia Hernández Zepeda, Dr. Jairo Cristóbal Alejo y Dr. Enrique Castaño de la Serna, por su dedicación durante la revisión de la presente tesis y las observaciones, las cuales enriquecieron el presente trabajo de investigación. A todos los que de una forma u otra han contribuido en mi formación académica y en la realización de la presente tesis de doctorado. DEDICATORIAS A todas las personas importantes de mi vida, por quienes soy lo que soy. ÍNDICE GENERAL RESUMEN ............................................................................................................................................. XVII ABSTRACT .......................................................................................................................................... XVIII INTRODUCCIÓN ....................................................................................................................................... 1 CAPÍTULO 1. ANTECEDENTES GENERALES .................................................................................... 3 1.1. CULTIVO DE PAPAYA .................................................................................................................. 3 1.2. DESCRIPCIÓN BOTÁNICA .......................................................................................................... 3 1.3. PRODUCCIÓN MUNDIAL Y NACIONAL DE PAPAYA E IMPORTANCIA ECONÓMICA ... 4 1.4. LIMITANTES FITOSANITARIAS DEL CULTIVO DE PAPAYA ................................................ 6 1.4.1. Papaya meleira virus (PMeV) .............................................................................................. 7 1.4.1.1. Sintomatología de la enfermedad pegajosa de la papaya ......................................................... 8 1.4.1.2. Diagnóstico del PMeV ...................................................................................................................... 9 1.4.1.3. Epidemiología del PMeV ............................................................................................................... 11 1.5. MECANISMOS DE TRANSMISIÓN DE VIRUS DE PLANTAS .............................................. 13 1.5.1. Transmisión por insectos vectores ................................................................................... 14 1.5.2. Transmisión no persistente ................................................................................................ 15 1.5.3. Transmisión persistente ..................................................................................................... 16 1.5.4. Transmisión semipersistente ............................................................................................. 17 1.5.5. Transmisión por mosquita blanca ..................................................................................... 17 1.5.6. Transmisión por áfidos ....................................................................................................... 18 1.5.7. Transmisión por chicharritas .............................................................................................. 21 1.6. JUSTIFICACIÓN .......................................................................................................................... 24 1.7. HIPÓTESIS ................................................................................................................................... 25 1.8. OBJETIVOS .................................................................................................................................. 26 1.8.1. General ................................................................................................................................. 26 1.8.2. Específicos ........................................................................................................................... 26 1.9. ESTRATEGIA EXPERIMENTAL ................................................................................................ 27 CAPÍTULO 2. EMPOASCA PAPAYAE (HEMIPTERA: CICADELLIDAE)-MEDIATED TRANSMISSION OF PAPAYA MELEIRA VIRUS IN MEXICO........................................................... 29 2.1. ABSTRACT ................................................................................................................................... 30 2.2. INTRODUCTION .......................................................................................................................... 31 2.3. MATERIALS AND METHODS .................................................................................................... 32 2.3.1. Insect’s collection ................................................................................................................ 32 2.3.2. Morphological and molecular identification of insects .................................................... 33 2.3.3. Screening of PMeV-Mx in insects by RT-qPCR and sequencing ................................ 34 2.3.4. Empoasca papayae colony establishment ...................................................................... 35 2.3.5. Plant material and virus source ......................................................................................... 35 2.3.6. Quantification of PMeV-Mx by RT-qPCR ......................................................................... 36 2.3.7. PMeV transmission tests .................................................................................................... 37 2.3.8. Acquisition of PMeV by E. papaya nymphs ..................................................................... 39 2.4. RESULTS ...................................................................................................................................... 39 2.4.1. Insect species diversity and abundance .......................................................................... 39 2.4.2. Detection of PMeV-Mx in insect species .......................................................................... 42 2.4.3. Establishing Empoasca papaya populations ................................................................... 43 2.4.4. Transmission of PMeV-Mx by Empoasca papayae ........................................................ 44 2.5. DISCUSSION ................................................................................................................................ 46 2.6. ACKNOWLEDGMENTS .............................................................................................................. 52 CAPÍTULO 3. MOLECULAR AND EXPERIMENTAL EVIDENCE OF OCCURRENCE OF MEXICAN VARIANT OF PAPAYA MELEIRA VIRUS IN COMMON WEED SPECIES IN PAPAYA ORCHARDS IN MEXICO ........................................................................................................................ 53 3.1. ABSTRACT ................................................................................................................................... 54 3.2. INTRODUCTION
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