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TESTING NATIVE ISOLATES of Cylindrobasidium Laeve for SUPPRESSION of REGROWTH of Acer Macrophyllum

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TESTING NATIVE ISOLATES of Cylindrobasidium Laeve for SUPPRESSION of REGROWTH of Acer Macrophyllum TESTING NATIVE ISOLATES OF Cylindrobasidium laeve FOR SUPPRESSION OF REGROWTH OF Acer macrophyllum Deepraj Purewal B.Sc. Ag. (Hons.), Punjab Agricultural University 1992 M.Sc. (Plant Pathology), Punjab Agricultural University 1995 THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF PEST MANAGEMENT In the Department of Biological Sciences O Deepraj Kaur Purewal2004 SIMON FRASER UNIVERSITY April 2004 All rights reserved. This work may not be reproduced in whole or in part, by photocopy or other means, without permission of the author. APPROVAL Name: Deepraj Purewal Degree: Master of Pest Management Title of Thesis: Testing native isolates of Cylindrobasidium laeve for suppression of regrowth of Acer macrophyllum Examining Committee: Chair: Dr. L. Lesack Dr. J.E, Rahe, Professor, Senior Supervisor Department of Biological Sciences, S.F.U. Dr. A.R. Kermode, Associate Professor Department of Biological Sciences, S.F.U. Dr. S.P. Lee, IT Systems Consultant Richmond, B .C. Dr. J.M. Webster, Professor Emeritus Department of Biological Sciences, S.F.U. Public Examiner Partial Copyright Licence The author, whose copyright is declared on the title page of this work, has granted to Simon Fraser University the right to lend this thesis, project or extended essay to users of the Simon Fraser University Library, and to make partial or single copies only for such users or in response to a request fi-om the library of any other university, or other educational institution, on its own behalf or for one of its users. The author has further agreed that permission for multiple copying of this work for scholarly purposes may be granted by either the author or the Dean of Graduate Studies. - It is understood that copying or publication of this work for financial gain shall not be allowed without the author's written permission. The original Partial Copyright Licence attesting to these terms, and signed by this author, may be found in the original bound copy of thn work, retained in the Simon Fraser University Archive. Bennett Library Simon Fraser University Burnaby, BC, Canada Abstract Bigleaf maple (Acer macrophyllum) produces large numbers of sprouts after the main stem is removed and the resultant large crowns interfere with growth of young conifers and constitute a hazard on hydroelectric rights of way. Management mainly relies on use of herbicides in combination with physical and manual control methods. However, recent research on preventing sprouting of stumps by inoculation with naturally occurring fungi has shown promise. The research described in this thesis was undertaken to evaluate the potential of Cylindrobasidium laeve for suppressing regrowth from cut stumps of A. macrophyllum. Three local isolates of C. laeve were applied to cut stumps of A. macrophyllum seedlings in June and September of 2001. Growth characteristics of isolates in axenic culture, and when inoculated into cut stems of selected native hardwood and softwood species were examined. Seedlings of A. macrophyllum treated in June of 2001 with two of the three isolates from culture media and all three isolates as colonized wood inoculum did not show any significant difference in number of shoots when compared with the negative control at the end of the nine weeks (P>0.05). Seedlings treated with one of these isolates from culture medium showed significant reduction in number of shoots as compared to the negative control treatment (P<0.05). However, all of the plants treated with C. laeve in June 2001 were alive and appeared healthy one year after treatment. None of the isolates suppressed regrowth of A. macrophyllum when applied in September of 2001. Growth of C. laeve on defined media was much less than on natural media. C. laeve grew comparably on three hardwood and three softwood species. Acknowledgements I express my deep sense of gratitude and extend thanks to my supervisor Dr. James E. Rahe for able guidance, invaluable suggestions and keen interest without which this work could not have been possible. I have real appreciation and regard for him for his unseizing encouragement. My sincere thanks are due to Dr. Stephen P. Lee and Dr. Allison R. Kermode, the members of my committee for their suggestions and critically going through the manuscript. It is my privilege to express my sincere gratitude to Dr. John M. Webster for being my public examiner. I wish to acknowledge the generous help that I received from Gwen Lohbrunner and Peter Hollman in the beginning of the project. My special thanks are due to Cheryl Calam, Arthur Moeller and Dave Trotter of the BC Ministry of Forests - Green Timbers Nursery for their help with the greenhouse experiments, Mohinder Singh Jawanda for providing greenhouse space when Green Timbers Nursery was shut down during the later stages of the experiment, Suchcha Singh Purewal, Malkiat Singh Hehar and Sunny for transporting plants from the greenhouse, Dr. Harry Kope for providing me with technical information and collecting the fungal isolates from Vancouver Island. I express my earnest thanks to the staff of BC Hydro SElP program (Gwen Shrimpton and Dr. Bob Bradley). I would like to thank Ian B. for his help with the statistical analysis of data. I'm thankful to Gagandeep Hehar for scanning, downloading and filing my research pictures. I offer my heartfelt appreciation to Bruce Leighton, Zaid Jumean, Christie MacDougall, and fellow graduate students for their friendship, feedback and encouragement, and the staff of the Department of Biological Sciences, especially Marlene Nguyen, Barbara Sherman, Brian Medford and Fiona for rendering ungrudging assistance whenever and wherever the need arose. From the conscience of my heart I feel a great pleasure to thank Tammy McMullan for her invaluable suggestions and emotional support all through my graduate studies at SFU. I express my earnest thanks to Dr. Santokh Singh Kang for sending me all the required documentation to get admission at SFU. Words at my command are not adequate to convey my profound gratitude to my affectionate husband, Budh Purewal for his constant, untiring and ever encouraging support and the self sacrifices will always remain indelible in my heart. It has indeed been a matter of pleasure to receive all possible help and moral support from Purewal and Hehar families especially from Gagandeep K. Hehar, which has enabled me to go through it all. I must pay my deepest and sincere thanks to the Almighty for steering me on the right path and enabling me to accomplish my task. Who waitedso long for this! andto the memory of my mother Whoinspiredme to continue my studies! Whoworfiedvery hardtogether to raise me! Their impvession on me w iKCast forever! Their unseCfirh he,their gentkness, their understandng, andtheir wkhm that shapedme into the person that I am today! Table of Contents Approval II Abstract iii Acknowledgements IV Dedication v Table of Contents vi .. List of Tables Vlll List of Figures ix 1.0. Introduction & Review of Literature 1 1.I. Weeds and woody plants in context 1 1.1.I. Biological control of weeds 3 1 .I.2. Biological control of undesirable vegetation in forestry 7 I.2. Bigleaf maple - Acer macrophyllum 9 1.2.1. Uses of Acer macrophyllum 10 1.2.2. Acer macrophyllum - Role in the ecosystem 10 1.2.3. Acer macrophyllum - A Weed 12 I.2.4. Control options for Acer macrophyllum 12 1.2.5. Fungi associated with Acer macrophyllum 14 1.3. Cylindrobasidium laeve - a potential mycoherbicide 16 1.3.1. Nomenclature and hierarchical classification of Cylindrobasidium laeve 19 1.3.2. Characteristics of Cylindrobasidium leave in axenic culture 2 1 1.3.3. Effect of Cylindrobasidium leave on regrowth of bigleaf maple 23 1.4. Objectives of the present research 25 2.0. Methods and Materials 26 2.1. Effect of Cylindrobasidium laeve on regrowth of cut stumps of Acer macrophyllum 26 2.1.1. Fungal isolates and fungal inoculum 26 2.1.2. Bigleaf maple trees 27 2.1.3. Experimental design treatments 27 2.1.4. Analysis of data 28 2.2. Growth characteristics of local isolates of Cylindrobasidium laeve in axenic culture 29 2.2.1. Growth on three undefined media 29 2.2.2. Effect of temperature and carbon source in defined media on rate of growth of Cylindrobasidium laeve 30 2.3. Growth of ~~lindrobasidiumlaeve in cut stem segments of selected native hardwood and softwood tree species 3 1 2.3.1 .Collection, inoculation and incubation of stem segments 3 1 3.0. Results 3.1. Effect of Cylindrobasidium laeve on regrowth of cut stumps of Acer macrophyllum 3. I.1. June - 2001 treatments 3.1.2. September - 2001 treatments 3.2. Growth characteristics of local isolates of Cylindrobasidium laeve in axenic culture 3.2.1. Effect of temperature on growth of Cylindrobasidium laeve 3.2.2. Growth on natural and defined media 3.3. Growth of Cylindrobasidium laeve on and in cut stem segments of selected native hardwood and softwood tree species 3.3.1. Growth of Cylindrobasidium laeve on outer surface of inoculated stem segments of selected native hardwood and softwood species at 5 "C, 15 OC and 22 OC 3.3.2. Growth of Cylindrobasidium laeve on the inner bark and cambial surfaces of wound inoculated stem segments of selected native hardwood and softwood species at 5"C, 15•‹Cand 22•‹C 4.0. Conclusions 45 4.1. Evidence that the Cylindrobasidium laeve has insufficient potential for biological control of resprouting of Acer macrophyllum 45 4.2. Conclusions regarding other aspects of research 47 4.3. Concluding remarks 48 References 5 1 vii List of Tables Table 1. Rating scale for fungal growth and discoloration of outer surface of inoculated cut stem segments. 32 Table 2.
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