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Diversity, Taxonomy and Phylogeny of Aristolochia Subsection Pentandrae (Aristolochiaceae) Thesis Presented By: Anna Paizanni Guillén

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Diversity, Taxonomy and Phylogeny of Aristolochia Subsection Pentandrae (Aristolochiaceae) Thesis Presented By: Anna Paizanni Guillén Universidad Michoacana de San Nicolás de Hidalgo Facultad de Biología PROGRAMA INSTITUCIONAL MAESTRÍA EN CIENCIAS BIOLÓGICAS MAESTRÍA EN ECOLOGÍA Y CONSERVACIÓN Diversity, taxonomy and phylogeny of Aristolochia subsection Pentandrae (Aristolochiaceae) Thesis presented by: Anna Paizanni Guillén In fulfillment of the requirements to obtain the degree of: Master in Biological Sciences Tutor: Dr. Juan Carlos Montero Castro (U.M.S.N.H.) Co-tutor: Dra. Marie-Stéphanie Samain (INECOL-Bajío) Morelia, Michoacán, March 2016 Universidad Michoacana de San Nicolás de Hidalgo Facultad de Biología PROGRAMA INSTITUCIONAL MAESTRÍA EN CIENCIAS BIOLÓGICAS MAESTRÍA EN ECOLOGÍA Y CONSERVACIÓN Diversidad, taxonomía y filogenia del género Aristolochia subsección Pentandrae (Aristolochiaceae) Tesis que presenta: Anna Paizanni Guillén Como requisito para obtener el grado profesional de: Maestra en Ciencias Biológicas Tutor: Dr. Juan Carlos Montero Castro (U.M.S.N.H.) Cotutor: Dra. Marie-Stéphanie Samain (INECOL-Bajío) Morelia, Michoacán, Marzo 2016 ACKNOWLEDGEMENTS The writing and completion of this thesis would not have been possible without the assistance, support and guidance of numerous people. I would like to thank all of them, namely the following. I want to thank the opportunity to realize this project in the Universidad Michoacana de San Nicolás de Hidalgo and the Centro Regional del Bajío of the Instituto de Ecología, A.C., as well as for the financial support by CONACYT and DAAD. To my essential support of life, my family and my life partner, to be with me unconditionally and for give me the motivation and encourangement during this work. My special thanks to my tutors and supervisor Dra. Marie-Stéphanie Samain, Dr. Juan Carlos Montero and Dr. Stefan Wanke to guide me during this thesis, introduce me in the molecular world, show me a different way to work more efficiently, provide financial support for the lab and fieldwork, and act during my thesis project as a supervisor. I´m very greatful to Dra. Deneb García Avila and Sabina Lara Cabrera for their contributions during the master period. I am especially indebted to my field group mates: to my life partner José Manuel Ramírez Amezcua, to my “nasty” friend Sebastian Müller, to Gaby Zacarias and Sarah Wagner, for helping me with enthusiasm, working with perseverance and motivation even in the hard days, and also for sharing many adventures during the field trips. Moreover, I´d like to thank some external people who helped during the search of pentandrous species: Eva Carolina, Evelyn Rios, Dr. Jacinto Treviño and Selene Rangel. Vielen Dank to the Plant Phylogenetics and Phylogenomics Research Group (Technische Universität Dresden, Germany) to introduce me to the molecular work and support me during my stay. Especially Dr. Stefan Wanke, M.C. Sebastian Müller and Markus Günther. I´m extremely greatful for your professional help. Furthermore, I want to thank Prof. Dr. Christoph Neinhuis, for his suggestions and his expert knowledge about the family Aristolochiaceae, which was very significant to me, knowing his point of view of Aristolochia. I would like to express my gratitude to all the curators of the following herbaria, for being so approachable during the revision of the collections: ANSM, Coahuila; ARIZ, Arizona, U.S.A.; BAYLU, Texas, Waco, U.S.A.; C, Copenhagen, Denmark; DR, Dresden, Germany; ENCB, Mexico City; GUADA, Jalisco; HCIB, Baja California Sur; XAL, Veracruz; MEXU, Ciudad de Mexico; NL, Nuevo León; QMEX, Querétaro; SLPM, San Luis Potosí; TEX, Texas, Austin U.S.A.; UCR, Riverside, California, U.S.A., and the Herbarium (ZEA), Jalisco. I am especially grateful to Dra. Socorro González, (CIIDIR), Geroge S. Hinton (GBH), and Dr. Sergio Zamudio (IEB), who facilitated the prospective trip and further supports. Furthermore, I would like to thank my botanical friends: Mollie Harker, Raymundo Ramírez Delgadillo, Jesús González Gallegos, and Arturo Castro Castro for encouraging my passion for botany and contribute with some Aristolochia sp. nov. inedit. collections; to Eduardo Ruíz Sánchez for his professional support; to Yocupitzia Ramírez Amezcua and Victor Steinmann for their kind support during the whole master period. Finally, I whish to express my gratitude to Francisco Javier Santana Michel, who introduced me to Aristolochia subsection Pentandrae; for his great knowledge in the field, and his contribution to this thesis. Dedicated to my family: Felipe, Lupis, Brenda, Felipe, Nicolás and my life partner Manuel Tablet of Contents RESUMEN ...................................................................................................................................... 1 ABSTRACT ..................................................................................................................................... 2 GENERAL INTRODUCTION AND OBJECTIVES ............................................................................... 2 CHAPTER 1. TAXONOMY OF ARISTOLOCHIA SUBSECTION PENTANDRAE ..................................... 6 1.1 INTRODUCTION ..................................................................................................................... 6 1.2 MATERIAL AND METHODS ..................................................................................................... 7 1.2.1. Databases, distribution maps and taxonomic treatment ............................................... 8 1.2.2. Fieldwork ..................................................................................................................... 8 1.3 RESULTS ............................................................................................................................... 9 1.3.1 Taxonomic treatment .................................................................................................. 10 1.3.1.1 Key to the species of Aristolochia subsection Pentandrae ................................... 17 1.3.1.2 Descriptions .......................................................................................................... 27 Aristolochia bracteosa .................................................................................................. 27 Aristolochia brevipes .................................................................................................... 29 Aristolochia cardiantha ................................................................................................ 31 Aristolochia coryi ......................................................................................................... 33 Aristolochia emiliae ...................................................................................................... 35 Aristolochia flexuosa .................................................................................................... 37 Aristolochia foetida ...................................................................................................... 38 Aristolochia karwinskii ................................................................................................. 42 Aristolochia lassa ......................................................................................................... 43 Aristolochia monticola ................................................................................................. 44 Aristolochia rzedowskiana ........................................................................................... 47 Aristolochia watsonii .................................................................................................... 49 Aristolochia sp. nov. inedit. (El Chavarín) ................................................................... 58 Aristolochia sp. nov. inedit. (Sierra Verde) .................................................................. 59 1.4 DISCUSSION ........................................................................................................................ 70 CHAPTER 2 PHYLOGENETIC RELATIONSHIP IN ARISTOLOCHIA SUBSECTION PENTANDRAE ..... 72 2.1 INTRODUCTION ................................................................................................................... 72 i 2.2. MATERIAL AND METHODS ................................................................................................. 77 2.2.1 Taxon Sampling and Plastid DNA sequences ............................................................ 81 2.2.2 DNA extraction, amplification and sequencing .......................................................... 86 2.2.3 Alignment and phylogenetic reconstruction ............................................................... 88 2.2.4. Nuclear loci ................................................................................................................ 89 2.2.4.1 The two selected nuclear loci ............................................................................... 89 2.2.4.2 Sampling and molecular work with nuclear single copy genes (NSCG). ............ 90 2.3 RESULTS ............................................................................................................................. 93 2.3.1 The plastid regions (DNA Chloroplast). ..................................................................... 93 2.3.2 Nuclear Single Copy Genes ........................................................................................ 96 2.4 DISCUSSION ........................................................................................................................ 97 2.5. GENERAL CONCLUSIONS ...................................................................................................
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