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Agaricomycetes of Ontario Tallgrass Prairies Western University Scholarship@Western Electronic Thesis and Dissertation Repository 8-15-2016 12:00 AM Agaricomycetes of Ontario Tallgrass Prairies Christopher R. J. Hay The University of Western Ontario Supervisor Dr. Greg Thorn The University of Western Ontario Graduate Program in Biology A thesis submitted in partial fulfillment of the equirr ements for the degree in Master of Science © Christopher R. J. Hay 2016 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Biodiversity Commons Recommended Citation Hay, Christopher R. J., "Agaricomycetes of Ontario Tallgrass Prairies" (2016). Electronic Thesis and Dissertation Repository. 3995. https://ir.lib.uwo.ca/etd/3995 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. Abstract This study provides the first characterization of the Agaricomycetes of Ontario tallgrass prairies, assesses the influence of various environmental factors, and compares results of aboveground mushroom surveys with belowground high-throughput DNA sequencing. Overall, the Mycenaceae, Ceratobasidiaceae and Polyporaceae were the most abundant, and the Clavariaceae, Entolomataceae and Sebacinaceae the richest in species. Position along a transect (geographic region) was the primary factor differentiating Agaricomycete composition of sites whereas tillage history and soil organic carbon content were secondary. The Hygrophoraceae and Clavariaceae were associated with pristine sites, and Minimedusa spp. associated with tillage. The belowground method captured most of the minor clades found aboveground and several more unique ones. The aboveground method retrieved 74 species and the belowground method 256 OTUs, with only eight shared between them. Keywords Agaricomycetes, tallgrass prairie, grassland, tillage, disturbance, restoration, soil organic carbon, fruiting bodies, mushrooms, soil fungal community, PCR, high-throughput sequencing, conservation, fungal biogeography, biodiversity, ribosomal DNA. i Acknowledgments Numerous people are acknowledged for their contributions to this project. First, a heartfelt thank-you to my supervisor, Dr. Greg Thorn. I am grateful for Greg’s support throughout my thesis: he accepted me as his student, guided me in the formation of this project, helped to provide conference and presentation opportunities, provided technical help in the lab and working with my data, and gave feedback during the final writing process. I also appreciate the guidance from my advisory committee: Dr. Zoë Lindo and Dr. André Lachance, especially Dr. Lindo’s assistance with R statistics and Dr. Lachance’s answers to questions related to phylogenetics and systematics. As my reader, Dr. Lachance provided the best grammatical and writing style feedback I have ever received. Thanks as well to Dr. Ben Rubin and Dr. Hugh Henry, whom I occasionally consulted for statistical or ecological questions, and Dr. Greg Gloor from biochemistry who provided essential code for my data pipeline and multivariate statistics. I thank all people who are a part of the Thorn lab for their assistance. I am indebted to Sarah Allan for providing me with step-by-step instruction in wet lab techniques and for the soil she collected in the field. Nimalka Weerasuriya assisted Sarah in soil collection and was indispensable to me during the bioinformatics pipeline stage and later to help with R code. Asma Asemaninejad gave her time to help me with PCR troubleshooting and generously shared her knowledge on appropriate methods of multivariate statistic techniques. Jennifer McDonald was always there when I had questions about anything – from technical details of my project to graduate student life. Catriona Catomeris and Aniruddho Chokroborty-Hoque also collected a share of the soil samples that were used in this project. Special thanks to previous lab members Barbara Bahnmann and Michael Lynch. Hard copies of their theses were always at hand as well- written examples. Thanks to the many work study students and lab volunteers who played a role in sequencing my mushroom specimens, especially my first and longest-lasting volunteer Linxi Xie, who was an exceptional assistant. I also thank my field site partners, without whom I would have no data: Clint Jacobs and our chaperones of Walpole Island First Nation; Andrea Zolnai, Meaghan ii Murphy, and Barbara Macdonell of the Herb-Gray Parkway project, as well as the MTO, MNRF, Parkway SAR team including Parkway Infrastructure Constructors (PIC), AMEC, and AECOM; Jim Wigle, the Ministry of Natural Resources and Forestry representative for Ojibway Prairie Provincial Nature Reserve; Bill Prieksaitis, director of the Elgin Stewardship Council, who permitted me to sample at Dutton-Dunwich prairie and provided directions and site details; Mary Garshore and Peter Carson, who showed me their prairie restoration site and allowed me to collect there; Jill Crosthwaite of the Nature Conservancy of Canada, who permitted me to sample at DeMaere prairie; and Jenna Quinn of the Rare Charitable Research Reserve, who was my contact for sampling at Blair Flats. Finally, thanks to my family and friends: my parents who helped me in the process of moving to Ontario, my sister and brother-in-law who were always available for conversation, and thanks for the support and fun times to the new friends I’ve made here – you know who you are. iii Table of Contents Abstract ................................................................................................................................ i Acknowledgments............................................................................................................... ii Table of Contents ............................................................................................................... iv List of Tables ................................................................................................................... viii List of Figures .................................................................................................................... ix List of Appendices ............................................................................................................. xi List of Site Abbreviations ................................................................................................. xii Chapter 1 - Introduction ...................................................................................................... 1 1 Introduction .................................................................................................................... 1 1.1 Agaricomycetes and their role in soils .................................................................... 1 1.2 Tallgrass prairies and agriculture ............................................................................ 2 1.3 Impacts to and conservation of soil fungi ............................................................... 4 1.4 Aboveground and belowground surveys of fungal diversity .................................. 7 1.5 Objectives ............................................................................................................... 9 1.6 Hypotheses and predictions .................................................................................... 9 Chapter 2 – Materials and Methods .................................................................................. 11 2 Materials and Methods ................................................................................................. 11 2.1 Site descriptions .................................................................................................... 11 2.1.1 Herb-Gray Parkway .................................................................................. 12 2.1.2 Ojibway Prairie Provincial Nature Reserve .............................................. 13 2.1.3 Walpole Island First Nation ...................................................................... 14 2.1.4 Dutton-Dunwich ....................................................................................... 15 2.1.5 Norfolk County ......................................................................................... 16 2.1.6 Blair Flats .................................................................................................. 17 iv 2.2 Field sampling ....................................................................................................... 17 2.2.1 Soil sampling design ................................................................................. 17 2.2.2 Mushroom collection ................................................................................ 19 2.3 Soil sieving............................................................................................................ 19 2.4 Molecular protocols .............................................................................................. 20 2.5 Mushroom identification ....................................................................................... 22 2.6 Vegetation metrics ................................................................................................ 22 2.7 Soil analyses.......................................................................................................... 24 2.7.1 pH .............................................................................................................
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