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Evaluación De Identificaciones Taxonómicas Mediante Código De Barras Del ADN En Un Grupo Tropical Megadiverso

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Evaluación De Identificaciones Taxonómicas Mediante Código De Barras Del ADN En Un Grupo Tropical Megadiverso Tesis Doctoral Evaluación de identificaciones taxonómicas mediante código de barras del ADN en un grupo tropical megadiverso Labarque, Facundo Martín 2012-04-11 Este documento forma parte de la colección de tesis doctorales y de maestría de la Biblioteca Central Dr. Luis Federico Leloir, disponible en digital.bl.fcen.uba.ar. Su utilización debe ser acompañada por la cita bibliográfica con reconocimiento de la fuente. This document is part of the doctoral theses collection of the Central Library Dr. Luis Federico Leloir, available in digital.bl.fcen.uba.ar. It should be used accompanied by the corresponding citation acknowledging the source. Cita tipo APA: Labarque, Facundo Martín. (2012-04-11). Evaluación de identificaciones taxonómicas mediante código de barras del ADN en un grupo tropical megadiverso. Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Cita tipo Chicago: Labarque, Facundo Martín. "Evaluación de identificaciones taxonómicas mediante código de barras del ADN en un grupo tropical megadiverso". Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. 2012-04-11. Dirección: Biblioteca Central Dr. Luis F. Leloir, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires. Contacto: [email protected] Intendente Güiraldes 2160 - C1428EGA - Tel. (++54 +11) 4789-9293 UNIVERSIDAD DE BUENOS AIRES Facultad de Ciencias Exactas y Naturales Departamento de Biodiversidad y Biología Experimental Evaluación de identificaciones taxonómicas mediante código de barras del ADN en un grupo tropical megadiverso Tesis presentada para optar al título de Doctor de la Universidad de Buenos Aires en el área Ciencias Biológicas Facundo Martín Labarque Director de Tesis: Martín Javier Ramírez Director Asistente: Miguel Ángel Arnedo Consejero de Estudios: Daniel Roccatagliata Lugar de trabajo: Museo Argentino de Ciencias Naturales “Bernardino Rivadavia” (CONICET), División de Aracnología. Buenos Aires, 6 de Febrero del 2012 i Evaluación de identificaciones taxonómicas mediante código de barras del ADN en un grupo tropical megadiverso Resumen El objetivo principal del proyecto fue evaluar las ventajas y limitaciones de los códigos de barras del ADN y de los métodos de la taxonomía clásica al estudiar la diversidad de arañas en los bosques montanos de Panamá. Se procesaron 29961 especímenes de arañas de 49 familias, de los cuales el 60% son juveniles, se secuenciaron 3553 fragmentos de cox1 y se generaron las bases de datos morfológica y molecular respectivas. Se identificaron los especímenes mediante tres estrategias concatenadas. Primero una identificación rápida utilizando caracteres somáticos externos, seguida por una identificación de grupos moleculares obtenidos con el modelo Generalizado Mixto de Yule y Coalescencia, finalizando con una identificación integrativa fruto de la comparación recíproca de las anteriores y del aporte de la disección de los órganos sexuales que son críticos a la hora de delimitar especies en artrópodos. Se identificaron 548 especies de manera integrativa, se estimó la diversidad alfa y beta según cada estrategia con los individuos adultos y, por primera vez, con los estadios inmaduros identificados molecularmente; se realizó una revisión taxonómica integrada de la familia Theridiosomatidae con el aporte de dos géneros nuevos y 27 especies nuevas y se asociaron los sacos de huevos con sus adultos mediante los códigos de barra, y se evaluaron las metodologías de colecta. Los códigos de barra sirvieron para estimar la diversidad, asociar diferentes estadios entre sí como machos y hembras y juveniles con sus adultos, revelar haplotipos endémicos, y sugirieron especies crípticas y nuevas especies putativas a partir de los estadios inmaduros no encontrados entre los adultos. Palabras clave: Diversidad, Sistemática, Arañas, Theridiosomatidae, Panamá ii Use of DNA barcoding techniques for rapid inventorying in a tropical megadiverse group Abstract The main goal of the project was to evaluate the advantages and limitations of DNA barcoding and traditional taxonomy to study the spider diversity in the montane forests of Panama. We have sorted 29961 specimens of 49 families, 60% of which were immature, and we have sequenced 3553 cox1 fragments, to generate morphological and molecular data bases. Specimens were identified using three linked approaches. First, a fast identification approach using external somatic characters, followed by a General Mixed Yule‐Coalescent molecular cluster identification, ending with the reciprocal illumination of both previous approaches plus a close examination of sexual organs, known as critic characters to delimit arthropods species, called integrative identification. We have identified 548 integrative species, we have estimated alpha and beta diversity with adults using each identification approach and, for the first time, we also have used immature specimens. We have done a taxonomic integrative revision of the family Theridiosomatidae describing two new genera and 27 new species plus spider egg‐sacs association with their species using DNA barcodes, and we have evaluated collecting methods. DNA barcodes can be used to estimate diversity, to associate different life and gender stages such as immature to mature and male to female specimens, and to reveal endemic haplotypes. DNA barcodes also suggest cryptic species, and putative new species from immature specimens not found among adults. Key words: Biodiversity, Systematics, Spiders, Theridiosomatidae, Panama iii ADVERTENCIA Esta Tesis no es válida como publicación, conforme al Capítulo 3 del CÓDIGO INTERNACIONAL DE NOMENCLATURA ZOOLÓGICA. Por lo tanto, los nombres nuevos y cambios taxonómicos propuestos aquí no tienen validez para fines nomenclatoriales o de orden de prioridad. WARNING This Thesis is not valid as publication, as described in the chapter 3 of the INTERNATIONAL CODE OF ZOOLOGICAL NOMENCLATURE. Therefore, taxonomic changes and new names proposed here are not valid for nomenclatural or priority purposes. ÍNDICE Resumen en castellano .................................................................................................... i Resumen en inglés (Abstract) ......................................................................................... ii Agradecimientos .............................................................................................................1 Dedicatoria ......................................................................................................................7 Un sueño zen ...................................................................................................................8 Objetivos Generales ........................................................................................................9 Introducción General ....................................................................................................11 La unidad básica ................................................................................................11 Antes de que se vayan .......................................................................................12 Una mano para la taxonomía ............................................................................13 Un nuevo vecino ................................................................................................. 15 Identificación de especímenes en BOLD ............................................................ 16 Enemigo íntimo ..................................................................................................17 Diversidad ..........................................................................................................19 Amigos de ocho patas ........................................................................................21 Identificación de especímenes ...........................................................................22 Materiales y Métodos ...................................................................................................25 Cuerpo de la Tesis ..............................................................................................25 Abreviaturas utilizadas ......................................................................................25 Sitios de estudio .................................................................................................27 Métodos de muestreo semi‐cuantitativos ......................................................... 27 Identificación de especímenes ........................................................................... 28 Códigos únicos de los especímenes ....................................................................29 Estimadores de diversidad .................................................................................32 Laboratorio taxonómico – Imágenes .................................................................35 Obtención del código de barras del ADN ...........................................................36 Análisis de las secuencias de ADN ......................................................................38 Capítulo 1: Rápido, rápido, rápido… Una aproximación integrativa a la estimación rápida de la diversidad de arañas Theridiosomatidas de los bosques montanos de la cordillera panameña .....................................................................................................42 Taxonomía ......................................................................................................... 42 Material examinado ...........................................................................................42
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