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Fusarium Head Blight of Wheat: Evaluation of the Efficacies Of Fusarium Head Blight of Wheat: Evaluation of the efficacies of fungicides towards Fusarium graminearum 3-ADON and 15-ADON isolates in spring wheat and assess the genetic differences between 3-ADON isolates from Canada and China By Chami Chathurangi Amarasinghe A Thesis Submitted to the Faculty of Graduate Studies In partial Fulfillment of the Requirements for the degree of MASTER OF SCIENCE Department of Plant Science University of Manitoba Winnipeg, Manitoba, Canada ©Copyright by Chami Chathurangi Amarasinghe 2010 THE UNIVERSITY OF MANITOBA FACULTY OF GRADUATE STUDIES ***** Fusarium Head Blight of Wheat: Evaluation of the efficacies of fungicides towards Fusarium graminearum 3-ADON and 15-ADON isolates in spring wheat and assess the genetic differences between 3-ADON isolates from Canada and China BY Chami Chathurangi Amarasinghe A thesis submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfillment of the requirements for the degree OF MASTER OF SCIENCE Chami Amarasinghe © 2010 Permission has been granted to the Library of the University of Manitoba to lend or sell copies of this thesis, to the National Library of Canada to microfilm this thesis and to lend or sell copies of the film, and to University Microfilm Inc. to publish an abstract of this thesis. The reproduction or copy of this thesis has been made available by authority of copyright owner solely for the purpose of private study and research, and may only be reproduced and copied as permitted by copyright laws or with express written authorization from the copyright owner. ACKNOWLEGMENTS First of all my sincere gratitude goes to Dr. W. G. D. Fernando, my supervisor, for his great support, guidance, encouragement and advice given to me throughout my Masters studies. He provided a valuable opportunity for me to be a part of his challenging research team. My heartiest gratitude goes to Drs Anita Brule Babel, Lily Tamburic-illinicic, and Jeannie Gilbert my advisory committee members, for the encouragement, help and invaluable guidance given to me throughout my research. They always helped me during my studies and gave me a lot of new ideas to improve my research. Special thanks to Ms. Paula Parks, for the great support given to me in the field, greenhouse and lab. Paula always helped me and guides me in every aspect. Also my heartiest gratitude goes to Ms. Mary Meleshko, for the invaluable help given to me for my field trials. I would like to thank Mr. Lyle Frisen, I really appreciate the help and advice given by you, for my data analysis. I would like to thank Nick and Mary, our summer students for helping me in the field. Also I would like to thank the staff in the general office, and green-house for their continuous help. I would like to convey my heartiest gratitude to my dear friends Ru Li and Dr. Khaled Al Taweel, for the invaluable support given to me. At last but not least, very special heartfelt thanks goes to my Amma, Thaththa, Nangi and Sajeewa for their invaluable never ending support and encouragement given to me throughout my Masters studies to make it a success. i ABSTRACT Amarasinghe, Chami Chathurangi. M.Sc., The University of Manitoba, December 2010. Fusarium Head Blight of Wheat: Evaluation of the efficacies of fungicides towards Fusarium graminearum 3-ADON and 15-ADON isolates in spring wheat and assess the genetic differences between 3-ADON isolates from Canada and China. Major Professor; Dr. W. G. D. Fernando. Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is one of the most destructive global diseases of small cereal grains worldwide. Severe epidemic outbreaks of FHB have been reported in North America, South America, Asia and Europe. The most devastating effect of this disease is the deposition of mycotoxins in the grain. Deoxynivalenol (DON) and its analogs 3-ADON, 15-ADON and NIV are the major mycotoxins produced by the fungus. The chemotype, 15-ADON is more prevalent in North America, but recently a chemotypic shift has been observed. The rapid emergence of the 3-ADON chemotype over the traditional 15-ADON chemotype is a major concern in North America. Understanding the behaviour of the 3-ADON chemotype and the reasons for the chemotypic shift is important in several aspects. In this study, the efficacy of fungicides in controlling FHB caused by 3-ADON and 15- ADON chemotypes was evaluated. All fungicides significantly reduced FHB disease symptoms and increased the yield, but some differing results were obtained from a few treatments. The findings of this study eluted to an integrated management approach over the use of a single management strategy to control FHB. This study evaluated the chemotype diversity of F. graminearum in corn, in Ontario and revealed that all investigated isolates were 15-ADON. Use of a single primer to differentiate 3-ADON, ii 15-ADON and NIV chemotypes is more efficient over multiplex polymerase chain reaction (PCR) approach. The reliability of using a single primer based on TRI13 gene to differentiate 3-ADON and 15-ADON chemotypes of F. graminearum was investigated. Interestingly, PCR based on TRI13 gene revealed genetic differences that exist between the 3-ADON chemotypes of Canada and China. The 3-ADON isolates from Canada had a 61 bp deletion in the TRI13 gene, but this deletion was absent in 3- ADON isolates from China. Therefore this confirms the presence of genetic differences among 3-ADON chemotypes based on their geographic origin. Another study revealed that the F. graminearum 3-ADON isolate can cause more disease over 15-ADON chemotype or from the combined infection of both isolates. iii FOREWORD This thesis is written in manuscript style. A general introduction and review of literature precedes manuscripts that comprised the main part of the thesis. Each manuscript consists of an abstract, introduction, materials and methods, results and discussion. A general discussion and conclusions, a list of references cited and a list of appendices follow the manuscripts. iv TABLE OF CONTENTS ACKNOWLEDGMENTS.............................................................................................i ABSTRACT....................................................................................................................ii FORWARD....................................................................................................................iii TABEL OF CONTENTS................................................................................................v LIST OF TABLES..........................................................................................................x LIST OF FIGURES.....................................................................................................xiv LIST OF APPENDICES...........................................................................................xviii CHAPTER 1 1.0 GENERAL INTRODUCTION................................................................................1 1.1 General introduction.................................................................................................2 CHAPTER 2 2.0 LITERATURE REVIEW........................................................................................5 2.1Wheat....................................................................................................................6 2.1.1 Origin of wheat................................................................................6 2.1.2 Classification and Types of Wheat..................................................6 2.1.3 Importance of wheat........................................................................9 2.1.4 Growth and Development of Wheat Plant.......................................9 2.1.5 Wheat production and trade...........................................................12 2.1.6 Wheat in Canada............................................................................12 2.2 Fusarium............................................................................................................14 2.2.1Origin and classification of Fusarium............................................14 2.2.2 Morphological characteristics and distribution of Fusarium spp.................................................................................15 2.2.3 Fusarium graminearum species....................................................16 v 2.3 Fusarium head blight....................................................................................18 2.3.1 Symptoms of FHB...............................................................19 2.3.2 FHB Disease cycle...............................................................19 2.3.3 Types of resistance against FHB development....................23 2.3.4 Infection of wheat by Fusarium..........................................23 2.4 FHB control strategies...................................................................................25 2.4.1 Cultural control......................................................................25 2.4.2 Biological control..................................................................27 2.4.3 Fungicide control...................................................................29 2.5 Breeding for FHB resistance........................................................................32 2.6Fusarium mycotoxins....................................................................................34 2.6.1 Trichothecenes.....................................................................34 2.6.2 Zearalenones........................................................................37
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