University of Texas at El Paso DigitalCommons@UTEP Open Access Theses & Dissertations 2012-01-01 Systematics of Leptopelis (Anura: Arthroleptidae) from the Itombwe Plateau, Eastern Democratic Republic of the Congo Francisco Portillo University of Texas at El Paso, [email protected] Follow this and additional works at: https://digitalcommons.utep.edu/open_etd Part of the Biology Commons, Developmental Biology Commons, Evolution Commons, and the Zoology Commons Recommended Citation Portillo, Francisco, "Systematics of Leptopelis (Anura: Arthroleptidae) from the Itombwe Plateau, Eastern Democratic Republic of the Congo" (2012). Open Access Theses & Dissertations. 1906. https://digitalcommons.utep.edu/open_etd/1906 This is brought to you for free and open access by DigitalCommons@UTEP. It has been accepted for inclusion in Open Access Theses & Dissertations by an authorized administrator of DigitalCommons@UTEP. For more information, please contact [email protected]. SYSTEMATICS OF LEPTOPELIS (ANURA: ARTHROLEPTIDAE) FROM THE ITOMBWE PLATEAU, EASTERN DEMOCRATIC REPUBLIC OF THE CONGO FRANK PORTILLO Department of Biological Sciences APPROVED: ______________________________ Eli Greenbaum, Ph.D., Chair ______________________________ Jerry D. Johnson, Ph.D. ______________________________ Rip Langford, Ph.D. ______________________________________ Benjamin C. Flores, Ph.D. Dean of the Graduate School Copyright © by Frank Portillo 2012 SYSTEMATICS OF LEPTOPELIS (ANURA: ARTHROLEPTIDAE) FROM THE ITOMBWE PLATEAU, EASTERN DEMOCRATIC REPUBLIC OF THE CONGO by FRANK PORTILLO, B.S. THESIS Presented to the Faculty of the Graduate School of The University of Texas at El Paso in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Department of Biological Sciences THE UNIVERSITY OF TEXAS AT EL PASO December 2012 ACKNOWLEDGMENTS First I would like to thank my family for their love and support throughout my life. As a child, my parents taught me how to appreciate wildlife and my love for the natural world began. I was always encouraged by my parents to learn and explore the outdoors without causing any disturbances to nature. My siblings have been great partners on many of my outdoor excursions and have always been a great support. My family continues to be a big inspiration to this day. I would like to thank my graduate committee, Dr. Eli Greenbaum, Dr. Jerry Johnson and Dr. Rip Langford. I would like to give a special thanks to my thesis advisor Dr. Eli Greenbaum who took me on as a student when I had very limited lab experience, and who provided superb guidance throughout my research. I would also like to thank all other faculty members that have helped along the way, including Dr. Elizabeth Walsh and Dr. Craig Tweedie for providing opinions. I also thank Dr. Vanessa Lougheed who carefully explained multivariate statistics. I thank Jens V. Vindum, Dr. David Blackburn (California Academy of Sciences) and Gregory Watkins-Colwell (Yale Peabody Museum) for access to specimens. DNA samples were sequenced by Omar Hernandez, Dr. Carolina Lema and Ana Betancourt at the UTEP DNA Analysis Core Facility (funded by a UTEP Master’s Thesis Grant). Additional funding was provided by the UTEP Graduate School, Department of Biological Sciences and College of Science. I am also thankful to lab members Chris Anderson who provided support and advice throughout the research process, Nancy Conkey and Fernie Medina for continual lab assistance and support throughout this project, and Katrina Weber and Cesar Villanueva for teaching me lab procedures when I was new. iv ABSTRACT Leptopelis, a genus of Central African treefrogs, includes 51 species that live in tropical forests and savannas. Currently, only two species of Leptopelis are known from the poorly explored Itombwe Plateau in eastern Democratic Republic of the Congo (DRC). Itombwe is renowned among conservationists for its rich and endemic amphibian fauna, including: Xenopus itombwensis, Chrysobatrachus cupreonitens, Laurentophryne parkeri, Hyperolius leleupi and at least three species of Arthroleptis. Evolutionary relationships of Itombwe Leptopelis were examined by sequencing two mitochondrial genes (16S: 557 bp [base pairs], cyt b: 620 bp) and one nuclear gene (RAG1: 761 bp). Results recovered strong support for several new lineages. Morphological characters and male advertisement call data were used to examine species boundaries in distinct lineages identified from the molecular phylogeny. Results indicated that the poorly known species Leptopelis fiziensis, originally described as a subspecies of L. modestus, is a distinct taxon from Cameroonian (topotypic) L. modestus, and a distinct east African population of L. modestus is described. Moreover a distinct L. karissimbensis population from Itombwe with low genetic divergence but distinct morphology and advertisement calls is described. The distinct calls between the recently diverged new species and the partially sympatric taxon L. karissimbensis are a likely consequence of reinforcement of species boundaries. This thesis represents the first phylogenetic analysis of Central African Leptopelis, and emphasizes the importance of the Itombwe Plateau as a conservation site. v TABLE OF CONTENTS ACKNOWLEDGMENTS ............................................................................................................... iv ABSTRACT ..................................................................................................................................v TABLE OF CONTENTS ................................................................................................................ vi LIST OF TABLES ...................................................................................................................... viii LIST OF FIGURES. ...................................................................................................................... Ix CHAPTER 1: INTRODUCTION ........................................................................................................1 1.1 Background .................................................................................................................1 1.2 Natural history ............................................................................................................4 1.3 Taxonomy and systematics .........................................................................................8 1.4 Study site ...................................................................................................................11 1.5 Research objectives ...................................................................................................17 CHAPTER 2: MATERIALS AND METHODS ..................................................................................19 2.1 Specimen acquisition ................................................................................................19 2.2 DNA extraction .........................................................................................................19 2.3 DNA amplification, sequencing and alignment ........................................................21 2.4 Phylogenetic analyses ...............................................................................................22 2.5 Population genetic analyses ......................................................................................23 2.6 Morphological analyses of adult frogs ......................................................................24 2.7 Morphological analyses of tadpoles..........................................................................25 2.8 Acoustic analyses ......................................................................................................26 CHAPTER 3: RESULTS ................................................................................................................28 3.1 Phylogenetic analyses ...............................................................................................28 3.2 Population genetic analyses ......................................................................................30 3.3 Itombwe Leptopelis morphology, coloration and calls .............................................37 vi 3.4 Description of Leptopelis anebos..............................................................................55 3.5 Description of Leptopelis versicolor .........................................................................62 CHAPTER 4: DISCUSSION ...........................................................................................................73 4.1 Taxonomic status and biogeography of Itombwe Leptopelis ...................................73 4.2 Conservation .............................................................................................................80 4.3 Conclusions and future directions .............................................................................82 LITERATURE CITED ...................................................................................................................83 APPENDIX I .............................................................................................................................101 APPENDIX II ............................................................................................................................105 VITA .......................................................................................................................................108 vii LIST OF TABLES Table 2.3.1