Phylogeography and Conservation Genetics of Two Endangered
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Phylogeography and Conservation Genetics of Two Endangered Amphibians, Blanchard's Cricket Frog (Acris crepitans blanchardi) and the Puerto Rican Crested Toad (Peltophryne lemur) Kaela B. Beauclerc A thesis submitted to the Committee on Graduate Studies in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Faculty of Arts and Science TRENT UNIVERSITY Peterborough, Ontario, Canada © Kaela B. Beauclerc 2009 Environmental and Life Sciences Ph.D. Program May 2009 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Abstract Phylogeography and Conservation Genetics of Two Endangered Amphibians, Blanchard's Cricket Frog (Acris crepitans blanchardi) and the Puerto Rican Crested Toad (Peltophryne lemur) Kaela B. Beauclerc I investigated the genetic diversity and structure of two endangered amphibians with the goal of developing conservation recommendations. The Puerto Rican crested toad (Peltophryne lemur) is a critically endangered tropical bufonid, for which one wild population remains. Declines are primarily due to habitat modification, and recovery efforts include captive breeding of northern and southern populations. In contrast, Blanchard's cricket frog (Acris crepitans blanchardi) is a temperate hylid that is declining at the northern edge of its range, but for which many large southern populations exist. The causes of declines are not well understood, and few conservation efforts have been initiated. I profiled the captive and wild populations of P. lemur, and 185 locations encompassing all Acris taxa, at the mitochondrial control region and several novel microsatellite loci. P. lemur had moderate microsatellite allelic diversity, but northern and southern populations were each fixed for a different mitochondrial haplotype. The northern population possessed many private microsatellite alleles that, when combined with habitat differences, may be indicative of adaptive variation. Given the highly inbred nature of the northern population and its likely extirpation, I recommend that a third breeding colony be established in which northern and southern individuals are mixed to preserve any unique northern traits. Profiling of A. c. blanchardi revealed a relatively Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. uniform northern clade and a substructured southern clade. Low genetic diversity in the north may be contributing to declines. The south was likely a refugium during Pleistocene glaciations, while the north experienced repeated population expansions and contractions. Phylogenetic analysis of all Acris species and subspecies indicated that substantial revisions to current taxonomy and distributions are required, including elevation of A. blanchardi to species status. Diversification within this complex primarily reflects large rivers, changes in elevation, and Pleistocene glaciations. Recognition of A. blanchardi as a species increases the urgency of conservation actions, and the distinctiveness of northern populations suggests that they may possess adaptive traits that should be conserved. Phylogeographic and spatial analysis provide a framework for actions such as reintroductions. Overall, this research demonstrates the importance of incorporating genetic information into recovery recommendations and management actions for threatened species. Keywords: amphibian, anuran, declining amphibian populations, endangered species, Acris, Peltophryne lemur, cricket frog, Puerto Rican crested toad, microsatellite, mitochondrial DNA, control region, phylogeography, genetic structure, conservation genetics, Pleistocene glaciation 11 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Acknowledgements Immense thanks go to my advisor, Dr. Brad White, for having the patience to see this through to the end. He provided the right amount of prodding, was always available for discussions, and could be counted on to keep the "bigger picture" in focus. I am also grateful to my committee members, Drs. Michael Berrill and David Galbraith, for providing a fresh perspective and forcing me to think outside the DNA. Bob Johnson at the Toronto Zoo is the inspiration for this work and life in general. His passion for these little frogs and their conservation is contagious. I am indebted to him for providing me with these fascinating projects, and for letting me tag along to Puerto Rico and participate in recovery efforts for the crested toad. This was full of unforgettable experiences, including swimming in the Caribbean Sea, maniac drivers, bomba with some locals, rainforests, tiny toad metamorphs, and roadside roasted chicken. Very special thanks go to Drs. Chris Wilson and Paul Wilson for countless chats about my research and my future. Dr. Tim Frasier cannot be thanked enough for his time explaining the minutiae to me and critiquing some of this work. Linda Cardwell of the ENLS program helped to smooth out the endless bumps encountered along the way. Her enthusiasm is refreshing. Andrew Lentini coordinated samples from the Toronto Zoo and endured many annoying emails from me. The cast and crew of the NRDPFC have made this experience enjoyable and unforgettable. Many technicians were instrumental in generating the data and I am sincerely grateful to all of them: Sue White-Sobey, Karen Mills, Kristyne Wozney, Smolly Coulson, and Jen Dart have variously sequenced, genotyped, and picogreened samples. Blane Bell is the automation wiz that maintained my sanity by extracting almost one thousand samples. Very special thanks go to my cohort of grad students, particularly Drs. Cathy Cullingham, Tim Frasier, Brenna McLeod, Roxanne Gillet, and Mark Ball for assistance, sympathy, rides to school, and life outside the lab. Dr. Stephen Petersen and his Mac were always ready to run ModelTest for me. 111 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Ariana Rickard and Edi Sonntag are thanked for dragging me along during their field work in Michigan so I could have some perspective on cricket frogs in real life. I am indebted to the following people for providing tissue samples of Acris: Joe Bartoszek, Neil Bass, Neil Bernstein, Will Bird, Luis Bonachea, Ron Bonnett, Jeff Briggler, Liz Burton, Brian Butterfield, Jeffrey Camper, Keith Coleman, Jeffrey Davis, Kevin Enge, Amy Estep, Eugenia Farrar, Dan Fogell, Carl Franklin, David Garcia, Dave Golden, Wayne Hildebrand, John Himes, John Jensen, Glen Johnson, Bob Jones, John Kleopfer, Kenny Krysko, Mike Lannoo, Jim Lee, Lisa Lehnhoff, Richard Lehtinen, Gregory Lipps, Mike Lodato, Malcolm McCallum, Brian Metts, Jonathan Micancin, Richard Montanucci, Nate Nazdrowicz, Jeff Parmelee, Steve Perrill, John Petzing, Stephen Richter, Ariana Rickard, Leslie Rissler, Floyd Scott, Don Shepard, Dirk Stevenson, David Swanson, Ashley Traut, Stan Trauth, and Zack Walker. Special thanks go to Donna Dittman at Louisiana State University Museum of Natural Science and Travis Taggart at the Sternberg Museum of Natural History, Fort Hays State University for arranging substantial tissue loans of Acris. Many thanks go to Miguel Canals of the Guanica State Forest and Rick Dietz of the Audubon Nature Institute for providing tissue samples of the Puerto Rican crested toad. Dr. Juan Rivero graciously allowed me to sample the historic specimens in his private collection, Jaime Matos-Torres provided additional specimens, and Gail Ross tromped around Puerto Rico collecting historic tissue samples, but these unfortunately were not incorporated. My family deserves special recognition for enduring me throughout my seemingly infinite university education. My parents, Murray and Marianne Jacobs, have encouraged and supported my education goals and love of biology and animals since I first protected that acre of rainforest in grade 9. Parker made sure I knew there was a world outside of my laptop and got some fresh air and exercise twice a day. My husband Michael encouraged, supported, listened, laughed, and vacuumed the house every week. He has seen me at my worst throughout this, and stuck it out anyways. He is my emotional stability and my best friend. Thank you for believing in me. IV Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Table of Contents Abstract ............................................................................................................................. ii Acknowledgements ........................................................................................................... iv List ofFigures .................................................................................................................. vii List of Tables ................................................................................................................. viii Chapter 1 General Introduction ......................................................................................................... 1 Declining amphibian populations ......................................................................... 1 Conservation genetics ........................................................................................... 3 Biology and conservation of Peltophryne lemur .................................................. 7 Biology and conservation of A eris crepitans blanchardi ................................... .14 Thesis objectives and overview