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Genetic Analysis of Resistance to Fusarium Head Blight in Wheat (Triticum Spp.)

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Genetic Analysis of Resistance to Fusarium Head Blight in Wheat (Triticum Spp.) Genetic analysis of resistance to Fusarium head blight in wheat (Triticum spp.) using phenotypic characters and molecular markers By Ali Malihipour A thesis submitted to the Faculty of Graduate Studies of the University of Manitoba in partial fulfilment of the requirements of the degree of DOCTOR of PHILOSOPHY Department of Biological Sciences University of Manitoba Winnipeg Copyright © 2010 by Ali Malihipour ABSTRACT Fusarium head blight (FHB), caused mainly by Fusarium graminearum (teleomorph: Gibberella zeae ), is one of the most damaging diseases of wheat. A ‘Brio’/‘TC 67’ spring wheat population was used to map quantitative trait loci (QTLs) for resistance to FHB, and to study the association of morphological and developmental characteristics with FHB resistance. Interval mapping (IM) detected a major QTL on chromosome 5AL for resistance to disease severity (type II resistance) and Fusarium-damaged kernels (FDK) under greenhouse and field conditions, respectively. Inconsistent QTL(s) was also detected on chromosome 5BS for disease severity and index using field data . The associations of plant height and number of days to anthesis were negative with disease incidence, severity, index, and deoxynivalenol (DON) accumulation data under field conditions. However, number of days to anthesis was positively correlated with disease severity (greenhouse) and FDK (field). Awnedness had a negative effect on FHB, namely the presence of awns resulted in less disease in the population. Spike threshability also affected FHB so that the hard threshable genotypes represented lower disease. Phylogenetic relationships of putative F. graminearum isolates from different sources were characterized using Tri101 gene sequencing data. Canadian and Iranian isolates clustered in F. graminearum lineage 7 (=F. graminearum sensu stricto) within the F. graminearum clade while the isolates received from CIMMYT , Mexico were placed in F. graminearum lineage 3 (= Fusarium boothii ) within the Fg clade or Fusarium cerealis . The PCR assay based on the Tri12 gene revealed the presence of the NIV, 3- ADON, and 15-ADON chemotypes with 15-ADON being the predominant chemotype. I While we did not find the NIV chemotype among the Canadian isolates, it was the predominant chemotype among the Iranian isolates. High variation in aggressiveness was observed among and within Fusarium species tested, with the isolates of F. graminearum sensu stricto being the most aggressive and the NIV chemotype being the least aggressive. The interactions between Fusarium isolates and wheat genotypes from different sources were investigated by inoculating isolates of F. graminearum sensu stricto and F. boothii on wheat genotypes. Significant differences were observed among the genotypes inoculated by single isolates. Results also showed significant interactions between Fusarium isolates and wheat genotypes. The F. boothii isolates from CIMMYT produced low disease symptom and infection on wheat genotypes regardless of the origin of the genotypes while F. graminearum sensu stricto isolates from Canada and Iran resulted in higher FHB scores. II ACKNOWLEDGEMENTS I gratefully acknowledge my supervisor Dr. Jeannie Gilbert for her valuable guidance, mentorship, and patience. Her strong support and continuous encouragement motivated me throughout my study leading to successful completion of the project. I extend my sincere appreciation to Drs. Anita Brûlé -Babel, George Fedak, and Michele Piercey-Normore for serving as my PhD committee members and for their valuable guidance, support, and the critical review of the thesis. I would like to thank Dr. Sylvie Cloutier (Cereal Research Centre, AAFC, Winnipeg, MB) for providing me access to her lab to do sequencing analysis of Fusarium isolates, her scientific advice, and comments on data analysis. I also thank Dr. Daryl Somers (Cereal Research Centre, AAFC, Winnipeg, MB) for providing the facilities for microsatellite analysis of the Triticum timopheevii derived mapping population in his lab I wish thank to Dr. Wenguang Cao (Eastern Cereal and Oilseed Research Centre, AAFC, Ottawa, Ontario) for development of the experimental wheat population, Dr. Sheila Woods (Cereal Research Centre, AAFC, Winnipeg, Manitoba) for her comments regarding experimental design in the greenhouse and field and data analysis, Dr. Kerry O’Donnell (National Center for Agricultural Utilization Research, USDA, Peoria, IL) for his information on PCR primers and Fg clade sequences, and Susan Patrick (Canadian Grain Commission, Winnipeg, Manitoba) for letting me access to her lab and assistance to do chemotype analysis of the Fusarium isolates. I would like to acknowledge Ron Kaethler, Kirsten Slusarenko, Uwe Kromer, Tim Unrau, Andrzej Walichnowski, Leslie Bezte, and Allison Brown for their kind cooperation and technical assistance in laboratory and field work at the Cereal Research III Centre, AAFC, Winnipeg, Manitoba. I also would like to thank Roger Larios (Dept. of Plant Science, University of Manitoba) for his assistance in Carman Fusarium nurseries and Sally Buffam (Eastern Cereal and Oilseed Research Centre, AAFC, Ottawa, Ontario) for DON analysis. I wish to thank all support and administration staff of the Cereal Research Centre, AAFC and Dept. of Biological Sciences, University of Manitoba for their kindness and help during my studies. My doctoral scholarship to do my PhD studies in Canada was funded by the Agricultural Research, Education and Extension Organisation (AREEO), Ministry of Agriculture, Iran which is greatly appreciated. Finally, I wish to thank my wife Mina Mahboubi and my children Parisa and Parsa for being supportive and patient during my studies and to my parents for their prayers and patience. IV To my wife Mina V TABLE OF CONTENTS ABSTRACT.....................................................................................................................I ACKNOWLEDGEMENTS ........................................................................................... III DEDICATION ............................................................................................................... V TABLE OF CONTENTS...............................................................................................VI LIST OF TABLES.........................................................................................................XI LIST OF FIGURES.................................................................................................... XIV INTRODUCTION........................................................................................................... 1 CHAPTER 1: GENERAL LITERATURE REVIEW....................................................... 5 Fusarium head blight................................................................................................... 6 Introduction............................................................................................................ 6 Symptoms .............................................................................................................. 8 The pathogens and geographical distribution .......................................................... 9 Epidemiology ....................................................................................................... 11 Disease cycle .................................................................................................. 11 Sources of inoculum ........................................................................................ 12 Inoculum production ....................................................................................... 14 Inoculum release and dispersal ........................................................................ 15 Infection and colonisation of the heads ............................................................ 17 Incubation and sporulation .............................................................................. 19 Sources of resistance ............................................................................................ 20 Components of resistance ..................................................................................... 26 VI Molecular and biochemical mechanisms of resistance........................................... 27 Inheritance of resistance ....................................................................................... 30 Pathogen profile ( Fusarium graminearum )................................................................ 34 Molecular phylogenetics and the Fusarium graminearum complex....................... 35 Genetic diversity of Fusarium graminearum populations...................................... 38 Mycotoxin production and trichothecene chemotypes........................................... 42 Variation in pathogenicity/aggressiveness............................................................. 47 Vegetative compatibility groups (VCGs) and phenotypic variation ....................... 48 Mapping of QTLs for fusarium head blight resistance ............................................... 49 Plant material ....................................................................................................... 49 Phenotyping ......................................................................................................... 50 Genotyping........................................................................................................... 52 QTLs for FHB resistance.....................................................................................
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