Duquesne University Duquesne Scholarship Collection Electronic Theses and Dissertations Spring 2010 Morphological and Genetic Investigations of Pennsylvania Populations of the Channel Shiner, Notropis wickliffi Stephanie Ann Dowell Follow this and additional works at: https://dsc.duq.edu/etd Recommended Citation Dowell, S. (2010). Morphological and Genetic Investigations of Pennsylvania Populations of the Channel Shiner, Notropis wickliffi (Master's thesis, Duquesne University). Retrieved from https://dsc.duq.edu/etd/500 This Immediate Access is brought to you for free and open access by Duquesne Scholarship Collection. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Duquesne Scholarship Collection. For more information, please contact [email protected]. MORPHOLOGICAL AND GENETIC INVESTIGATIONS OF PENNSYLVANIA POPULATIONS OF THE CHANNEL SHINER, NOTROPIS WICKLIFFI A Thesis Submitted to the Bayer School of Natural and Environmental Sciences Duquesne University In partial fulfillment of the requirements for the degree of Master of Science By Stephanie A. Dowell May 2010 Copyright by Stephanie A. Dowell 2010 MORPHOLOGICAL AND GENETIC INVESTIGATIONS OF PENNSYLVANIA POPULATIONS OF THE CHANNEL SHINER, NOTROPIS WICKLIFFI By Stephanie A. Dowell Approved March 10, 2010 ________________________________ ________________________________ Dr. Brady Porter Dr. Kyle Selcer Associate Professor of Biology Associate Professor of Biology (Committee Chair) (Committee Member) ________________________________ Dr. Lisa Ludvico Assistant Professor of Biology (Committee Member) ________________________________ ________________________________ Dr. David W. Seybert Dr. Phillip Auron Dean, Bayer School of Natural and Chair, Department of Biological Sciences Environmental Sciences Professor of Biology iii ABSTRACT MORPHOLOGICAL AND GENETIC INVESTIGATIONS OF PENNSYLVANIA POPULATIONS OF THE CHANNEL SHINER, NOTROPIS WICKLIFFI By Stephanie A. Dowell May 2010 Thesis supervised by Dr. Brady A. Porter The channel shiner, Notropis wickliffi, is listed as a vulnerable species in Pennsylvania. Little is known about this species of minnow due to taxonomic confusion with the mimic shiner, N. volucellus. The regional variation in morphology makes it problematic to apply identification characters determined in other studies to Pennsylvania populations. By sequencing the mitochondrial gene, cytochrome b, it was found that N. wickliffi formed a separate phylogenetic clade, while N. volucellus and N. buchanani (ghost shiners) were present in the same clade. Three pigmentation patterns were correlated to the haplotype groups, proving useful in distinguishing between N. wickliffi and N. volucellus from Pennsylvania. Principal component analysis revealed that the morphology of all three species is statistically different. Eight polymorphic microsatellite loci, analyzed with STRUCTURE, showed that hybridization is most likely taking place iv between both N. wickliffi and N. volucellus and also between N. buchanani and N. volucellus. v DEDICATION I would like to dedicate this work to my parents, John and Kathy Dowell, for their endless support and encouragement. vi ACKNOWLEDGEMENT I would like to thank my advisor, Dr. Brady Porter, for all of his help and encouragement. This experience has taught me more than simply laboratory techniques, but valuable skills that will last a lifetime. His patience coupled with his excitement for science and teaching made for the perfect learning environment. Thank you so much for providing me with so many wonderful opportunities. Funding for this research was mainly provided by the Pennsylvania Department of Conservation and Natural Resources: Environmental Stewardship Fund - Wild Resource Conservation Program. Additional funding as well as the main source of specimens came from the Pennsylvania Department of Environmental Protection as part of the Environmental Protection Agency’s Environmental Monitoring and Assessment Program – Great Rivers Ecosystem Project. Specifically, I would like to thank Rick Spear, Gary Kenderes, and Dan Counahan with the Pennsylvania DEP, and Dr. Elizabeth Dakin and Laura Howell from Duquesne University for helping with the sample collection. Specimens were also obtained with the assistance of Doug Locy with Aquatic System’s Incorporated. Also, the Ohio specimens were collected with the help of Marc Kibbey from Ohio State University. His assistance was definitely appreciated. The morphological reference specimens were provided by the Ohio State University Museum Fish Division under the direction of Ted M. Cavender and Marc R. Kibbey. I would like to thank Dr. Jay Stauffer from Penn State University for taking the time to patiently run and explain principal component analysis to me. These results were invaluable to this project and could not have been produced without his assistance. vii Thanks to Robert A. Hrabik with the Missouri Department of Conservation for providing me with his Master’s thesis on N. wickliffi and N. volucellus in the Mississippi River. He also provided feedback for this project that was much appreciated. Dr. Douglas Landsittel and Dr. John Kern from Duquesne University advised me as to the statistical methods to use for the pigment combination study. Other Porter Lab members, such as Maria Wheeler, Katie Boone, Melanie Mathews, Ben Latoche, Alexis Smith, and TJ Fernino created a supportive and fun work environment. This experience would not have been as enjoyable without them! Finally, I would like to thank Duquesne University’s Bayer School of Natural and Environmental Sciences for supporting me for on a teaching assistantship and for making this research possible. viii TABLE OF CONTENTS Page ABSTRACT....................................................................................................................... iv DEDICATION................................................................................................................... vi ACKNOWLEDGEMENT ................................................................................................ vii LIST OF TABLES............................................................................................................. xi LIST OF FIGURES .......................................................................................................... xv CHAPTER 1: BACKGROUND........................................................................................ 1 Biology............................................................................................................................ 2 Identification Problems................................................................................................... 8 History of the Allegheny and Monongahela Rivers ....................................................... 9 CHAPTER 2: MITOCHONDRIAL INVESTIGATION................................................ 11 Introduction................................................................................................................... 11 Methods......................................................................................................................... 15 Sample Collection..................................................................................................... 15 Genetic Analysis ....................................................................................................... 19 Results........................................................................................................................... 27 CHAPTER 3: MORPHOLOGICAL INVESTIGATION............................................... 39 Introduction................................................................................................................... 39 Methods......................................................................................................................... 42 Pigmentation ............................................................................................................. 43 Morphometrics and Meristics ................................................................................... 45 Results........................................................................................................................... 49 ix Pigmentation ............................................................................................................. 49 Morphometrics and Meristics ................................................................................... 56 Discussion..................................................................................................................... 71 Pigmentation ............................................................................................................. 71 Morphometrics and Meristics ................................................................................... 74 CHAPTER 4: MICROSATELLITE INVESTIGATION ............................................... 80 Introduction................................................................................................................... 80 Methods......................................................................................................................... 82 Results........................................................................................................................... 87 Discussion................................................................................................................... 102 CHAPTER