University of Alberta Enhanced Revegetation and Reclamation of Oil Sand Disturbed Land Using Mycorrhizae. By Nnenna Esther Onwuchekwa A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfilment of the requirements for the degree of Master of Science In Land Reclamation and Remediation Department of Renewable Resources © Nnenna Esther Onwuchekwa Spring 2012 Edmonton, Alberta Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. 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ABSTRACT This study examined the potential of using conifer seedlings Picea glauca and Pinus banksiana inoculated with ectomycorrhizal (ECM) fungi to improve revegetation success and plant establishment in reclaimed oil sands mining sites. Mycorrhizal inoculum potential of the reclamation soils was low with the maximum inoculum potential of 23% and 29%for ECM and arbuscular mycorrhizae, respectively. The response of seedlings in the field to ECM inoculation varied between plant species and measured parameters. A significant effect of ECM inoculation on height was observed in P. banksiana but not in P. glauca. The average survival rate for P. glauca seedlings inoculated with different species of ECM varied between 36% and 56%, whereas the control (uninoculated) seedlings had minimum and maximum survival rates of 22 and 41% respectively. Generally, it was construed that the re-introduction of mycorrhizal fungi during reclamation process is an important approach that should be further exploited. ACKNOWLEDGEMENT My sincere gratitude goes to my supervisor Janusz Zwiazek whose thorough guidance, corrections and objective criticisms contributed to the improvement of my research and writing skills. Above all I appreciate him for accepting me as a graduate student under his supervision for which this thesis completion would not be realized. My sincere appreciation goes also to Dr. Ali Quoreshi for his unending efforts and guidance during my field work. Indeed, you became not only a mentor but a friend. My profound gratitude goes to Suncor Energy, Inc. for providing the funds for this project. To Francis Salifu who made sure we had all the necessary help on site as well as other Suncor Staff, your assistance was greatly valued. Charles Greer (NRC Montreal), Alain Atangana (U of A), Rasheed Adeleke (South Africa) and Gaetan Daigle (U of Laval), their involvement is gratefully acknowledged. Many thanks to my Lab colleagues who gave suggestions during lab meetings or experiments, your comments were appreciated. I am indeed very thankful to my supervisory committee, Scott Chang and Damase Khasa whose academic inputs improved the quality of the thesis. Great thanks to the University of Alberta, University of Laval, 2020Seedlab and Symbiotech for the in-kind support provided in the course of this degree. Also, to my friends Ayoola, Christelle, Godwin, Adaku, Uche etc in Edmonton and family members : Mum, Nkechi, Linda and Emeka Ike in Nigeria and Ontario, the moral support, fun and laughter we shared was indeed heart- warming. Finally, to my husband Kingsley Onwuchekwa and Son Israel who supported me and took time out to enable me focus during the final stages of the thesis, I love you and you are intensely appreciated. TABLE OF CONTENTS 1 LITERATURE REVIEW ........................................................................ - 1 - 1.1 Introduction .................................................................................................................... - 1 - 1.2 Oil Sands mining ............................................................................................................. - 1 - 1.3 Oil Sands reclamation and revegetation ......................................................................... - 4 - 1.4 Application of mycorrhizal biotechnology in reclamation ............................................. - 6 - 1.4.1 Mycorrhizas and benefits .............................................................................................. - 6 - 1.4.2 Possible factors that can affect mycorrhizal establishment in reclamation ....................... - 9 - 1.4.3 Reclamation of disturbed lands using ectomycorrihzal fungi .......................................... - 9 - 1.5 Ectomycorrhizal fungi and their eological importance .................................................- 17 - 1.6 Mycorrhizal research application in Canada ................................................................- 17 - 1.7 Motivation for the study ................................................................................................- 22 - 1.8 Objectives and thesis structure ......................................................................................- 22 - 1.9 Literature cited ..............................................................................................................- 23 - 2 MYCORRHIZAL INOCULUM STATUS OF RECLAMATION SOIL SUBSTRATES FROM THE ATHABASCA OIL SANDS REGION .............. - 35 - 2.1 Introduction ...................................................................................................................- 35 - 2.2 Materials and Methods ..................................................................................................- 37 - 2.2.1 Soil Substrate ...............................................................................................................- 37 - 2.2.2 Seedling Preparation ....................................................................................................- 37 - 2.2.3 Bioassay Setup.............................................................................................................- 38 - 2.2.4 Morphological description and molecular identification of ectomycorrhizal fungi .........- 39 - 2.2.5 Data Analysis ..............................................................................................................- 40 - 2.3 Results ............................................................................................................................- 41 - 2.3.1 Seedling growth ...........................................................................................................- 41 - 2.3.2 Morphotyping and mycorrhizal status ...........................................................................- 42 - 2.4 Discussion .......................................................................................................................- 45 - 2.5 Literature cited ..............................................................................................................- 48 - 3 ECTOMYCORRHIZAL BIOTECHNOLOGY: AN APPLICATION TO ENHANCED REVEGETATION AND RECLAMATION OF OIL SANDS DISTURBED LANDS. ...................................................................................... - 54 - 3.1 Introduction ...................................................................................................................- 54 - 3.2 Materials and Methods ..................................................................................................- 57 -