Potential genetic impacts of metal content on Northern Red Oak (Quercus rubra) populations from the Greater Sudbury Region during reclamation By Anh Tran Thesis submitted in partial fulfillment of the requirements for the degree of Master of Science (MSc) in Biology School of Graduate Studies Laurentian University Sudbury, Ontario © Ann Tran, 2013 Library and Archives Bibliotheque et Canada Archives Canada Published Heritage Direction du 1+1Branch Patrimoine de I'edition 395 Wellington Street 395, rue Wellington Ottawa ON K1A0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre reference ISBN: 978-0-494-91872-2 Our file Notre reference ISBN: 978-0-494-91872-2 NOTICE: AVIS: The author has granted a non­ L'auteur a accorde une licence non exclusive exclusive license allowing Library and permettant a la Bibliotheque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par telecommunication ou par I'lnternet, preter, telecommunication or on the Internet, distribuer et vendre des theses partout dans le loan, distrbute and sell theses monde, a des fins commerciales ou autres, sur worldwide, for commercial or non­ support microforme, papier, electronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. 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Canada Abstract Genetic diversity is important for species survival and sustainability under environmental stresses and changes. The objectives of the study are to compare genetic variability in red oak populations growing in eroded and stable upland sites and to determine if there are any long term effects of liming on aspects of soil fertility and toxicity, and population variability of red oak. Total and bioavailable metals were measured in samples from different sites within the Greater Sudbury Region. The highest concentrations of metals were found in the upper organic layer, LFH. Only a small fraction of total metals were bioavailable. The level of metal content in the top soil layer was affected by both land topography and the site distance from smelters. The enrichment factor comparing the contaminated versus uncontaminated sites varied between 0.702 and 16.78 indicating environmental pollution. The translocation factors from soil to branches were low for most total elements, but high for bioavailable metals. The pH levels of limed areas were found to be higher than unlimed areas, an indication of the prolonged effect of liming on soil acidity. Genetic analysis of all the red oak populations was conducted using ISSR markers. The level of polymorphic loci in red oak populations from the Greater Sudbury Region was moderate to high ranging from 43.97% to 64.54%. The mean values of Na, Ne, h and I were determined to be 1.54, 1.22, 0.14 and 0.22, respectively. The levels of inter-population polymorphism and population differentiation (G s t ) were 97.87% and 34%, respectively. The estimated gene flow (Nm) value was 0.98. The genetic distance values ranged between 0.125 and 0.611. There was no association between the level of soil metal content and the genetic variation of the red oak populations analyzed. iv Acknowledgements I would like to take this opportunity to sincerely thank my supervisor Dr. Nkongolo for giving me the opportunity to work in his lab and for his dedication, passion and expertise throughout my research project. To my committee members Dr. Spiers and Dr. Beckett for their assistance with site selection, sample collection and metal analysis. To my labmates Ramya Narendrula, Paul Michael, Melanie Mehes and Gabriel Theriault for their valuable discussions and assistance with sample collection, processing and data analysis. I would like to express appreciation to members of the Elliott Lake Research Field Station of Laurentian University analysts for metal analysis. I would like to thank my loving parents for their continuous support and believing in me, as well as my friend Martin Uceda for his motivation when I needed it most. Table of Contents ABSTRACT....................................................................................................................................................... III ACKNOWLEDGEMENTS..............................................................................................................................V LIST OF FIGURES........................................................................................................................................ VIII LIST OF TABLES...............................................................................................................................................X CHAPTER 1: LITERATURE REVIEW.......................................................................................................... 1 1 .1 M etal C ontamination in t h e G r e a ter S u d b u r y R e g io n ( O n ta rio , C a n a d a ) ............................. 1 1 .2 N u t r ie n t C o n t e n t in S o il ............................................................................................................................................. 3 1 . 3 S o il A m e n d m e n t ...................................................................................................................................................................8 1 .3 R evegetation .....................................................................................................................................................................1 0 1 .4 S pe c ie s o f In t e r e s t .......................................................................................................................................................... 1 2 1 .5 O v e r v ie w o f G en etic variability ........................................................................................................................... 1 5 1 .6 .1 M e a su r in g G en etic variability w it h M o l ec u la r M a r k e r s .................................................................1 5 1 .6 .2 In t e r -S im p l e S e q u e n c e R e p e a t (ISSR)-PCR ...................................................................................................1 8 1 .6 O b je c t iv e s ............................................................................................................................................................................ 2 0 CHAPTER 2: METAL ANALYSIS IN RED OAK (QUERCU5 RUBRA) POPULATIONS GROWING IN LIMED AND UNLIMED AREAS IN THE GREATER SUDBURY REGION (ONTARIO, CANADA)................................................................................................................................. 21 2 .1 Introduction ........................................................................................................................................................................2 1 2 .2 M ateria ls a n d M e t h o d s ............................................................................................................................................... 2 2 2 .2 .1 S a m p l in g ..............................................................................................................................................................................2 2 2 .2 .2 M etal a n d p h a n alysis in s o i l ................................................................................................................................2 5 2 .2 .3 S ta tistic a l A n a l y s is ..................................................................................................................................................... 2 6 2 .3 R e s u l t s ..................................................................................................................................................................................... 2 7 2 .3 .1 T o t a l m e t a l o f S o i l ..................................................................................................................................................... 2 7 2 .3 .2 B ioavailable M etal o f S o i l .................................................................................................................................... 2 8 2 .3 .3 S o il ph lev els .................................................................................................................................................................... 2 9 2 .3 .4 E n r ic h m e n t Fa c t o r ..................................................................................................................................................... 2 9 2 .3 .5 T ranslocation f a c t o r ..............................................................................................................................................