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This reproduction is the best copy available UMI MITOCHONDRIAL DNA DIVERSIT'Y AND ANTHROPOGENIC INFLUENCES IN WALLEYE (Stizostedion vitrem) FROM EASTERN LAKE HURON A Thesis Presented to The Facdty of Graduate Studies of The University of Guelph by MICHAEL HENRY GATT In partial fulfifment of requirements for the degree of Master of Science December 1998 @ Michael Henry Gatt, 1998 Natidnal Library BiMiièque nationale du Canada Acquisitions and Acquisitions et Bibliographie Services seMces bibliographiques 395 Wellmgtm Street 305. rue WMnm ûüawaON K1AW OttawaON K1AW Canada Canada The author has granted a non- L'auteur a accordé une licence non exclusive licence allowing the exclusive permettant à la National Lrtbrary of Canada to Bibliothèque nationale du Canada de reproduce, loan, distriiute or seLl reproduire, prêter, disîribner ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or elecironic formats. la forme de microfiche/nIm, de reproduction sur papier ou sur format éIectrooique. The author retains ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni la thèse ni des extraits substantiels may be printed or otherwise de celle-ci ne doivent être imprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation. MITOCHONDRIAL DNA DIVERSITY AND ANTHROPOGENIC INFLUENCES IN WALLEYE (Stizostedion FROM EASTERN LAKE HURON Michael Henry Gatt Advisor: University of Guelph, 1998 Professor M.M. Ferguson MtDNA diversity in contemporary samples from eight populations of walleye kom eastem Lake Huron was negatively associated with the relative amounts of anthropogenic impacts experienced by each. MtDNA diversity was also compared in contemporary and archival samples from two populations. Ln one instance, contemporary samples had reduced mtDNA diversity compared to archival samples collected before major exploitation. No such difference was detected between contemporary and archival samples fiom the second population where commercial exploitation and habitat alteration have been more limitecl. Law mtDNA diversity in artificially produced fish relative to parental sources suggests that culture programmes might be one mechanism lowering mtDNA diversity in populations. A phylogenetic cornparison of walleye mtDNA haplotypes revealed thai an analysis based on restriction fkagment length polymorphisms in two amplified fÎagments surveyed 554bp and detected almost twice the number of mutations and haplotypes than did a sequence analysis of 5 13nt of the control region. Foremost, I thank Dr. Moira Ferguson for her advice and guidance during my thesis program. I am especially thankfiil to both Drs. Moira Ferguson and Roy Dari~nannfor providing me with the opportunity to partake Ui intereshg research in their lab at the University of Guelph. Their direction and encouragement over the years is greatly appreciaîed. Drs. Roy Danzmann and Teri Crease served as comIILittee mernbers and offered constructive crïticism and insight with various facets of the study for which I am grateful. 1owe special thanks to Amas Liskauskas (Management Biologist - Lake Huron Management Unit) who graciously arranged bding for this study. This research was supported by hdslkm the Lake Huron Mauagement Unit, Ontario Ministry of Natural Resources, and a Fisheries Research Grant fkom the Ontario Federation of Anglers and Hunten. Aninas Liskauskas also deserves thanks for coordinating field collections and providing various walleye samples. Help with collecting additional samples was provided by personnel with the Ontano Ministry of Natural Resources: Jeff Buck, Steve Gile, Butch Lafiance, Eric Machtyre, Wayne Sehger, Jerry Srnitka, and Bruce Stedmen. 1 thank Eric Machlyre and George Morgan (Department of Biobgy, Laurentian University) for locating and providing archival scale samples. Lamy Wickett (Aqua Tem Contracting) is thanked for his help with collecting samples of walleye hm the Moon Riva. 1 also wish to aclmowledge Robert Caphg, Chuck Frail, Al Hoosen, Rob Linder, Dave Russell, Jerry Smitka, and Rino Ouellette for helping obtain samples of adults from various populations and for obtaining pond-reared walleye. Dr. Neil Billington (Zoology Department, Southern Illuiois University) generously provided purified mtDNA of a divergent walleye haplotype hma Mississippian drainage in the USA. Also, 1 appreciate the time and help offered by Tara McParland. Tara provided partially purified mtDNA from numerous walleye samples collected during her M.Sc. thesis research, Usefiil discussions concerning the technical and molecular analyses of my work occwed with several people sharing the lab. In particular, 1 appreciate insight fiom Tony Fishback, David Gislason, Guy Perry, Ana Rakitin, Takashi Sakamoto and Rachael Woram. They provided practical advice when troubleshooting was necessary. 1 particularly enjoyed partaking in philosophical conversations with my lab mates at the Graduate Lounge on Friday evenings! John Colbourne and Kerry Naish provided assistance with the phylogenetic analyses and Ana Rakitin provided useful comments on previous versions of the thesis. 1thank Angela Holliss for sequencing amplified fragments fkom my walleye mtDNA samples. Ian Smith deserves many thanks for his help and time with bioimagixg Lastly, 1 thank rny family and Een& for their understanding and support while 1 conducted my MSc. research. Mostly, 1thank Tiffany Hele for her votes of optimism and overwhelrning encouragement while 1completed my thesis over the last year. TABLE OF CONTENTS ... TAE3LE OF CONTENTS ............................................................................................................... iii LIST OF TABLES ......................................................................................................................... iv LIST OF FIGURES ............. .. ...................................................................................................... v MATERIALS AND METHODS .................................................................................................... 8 F'HYLOGENETIC ANALYSIS OF MTDNAHAPLOTYPES ................. ......... ........................................ 8 Samples .................................................................................................................................... 8 DNA isolation .......................................................................................................................... 8 .......,..... Sequence analysis ............................ ,.,. .......,..... 9 Fragment-RFLP anaiysis ........................................................................................................ 10 mtDNA variation detected by sequence and fragment-RFLP analyses ................................. 12 Relationships among haplotypes ............................................................................................ 13 MTDNA DlVERsITY AND EXPLOITAïION HISTORY .................................................................... 15 Samples .................................................................................................................................. 15 DNA isolation ........................................................................................................................ 16 PCR of contmporary samples ............................................................................................... 16 PCR of archival samples ........................................................................................................ 16 Statisticai anaiysis .................................................................................................................. 17 PHYLOGENETICANALYSE OF MTDNAHAPLOTYPES ................................................................ 19 Sequence analysis ................................................................................................................ 19 Fragment-RFLP analysis........................................................................................................ 20 mtDNA variation detected by sequmce and hgmmt-RFLP analyses ............................ ..... 20 Relationshrps among haplotypes ............................................................................................ 21 MTDNA DIVERsïW AND EXPLOITATION HISTORY................................................................ 23 Contemporary distribution of mtDNA haplotypes ........................... .. .................................... 23 Historical distribution of mtDNA haplotypes ........................................................................ 24 mtDNA distribuîions in cultured fish and correspondmg source populations ....................... 24 DISCUSSION ...................... .. ....................................................................................................... 40 PHYLoGENETIC ANALYSIS OF MTDNAHAPLOTYPES ................................................................ 40 mtDNA variation detected by sequence and ftagment-RFLP analyses ................................. 40 Relationships among haplotypes ............................................................................................ 43 MTDNADIVERSITY AND EXPLOITATION HISTORY ...................................................................