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Identify New Resistant Genes for Eyespot Diseases of Wheat In IDENTIFICATION AND MAPPING OF RESISTANCE GENES FOR EYESPOT OF WHEAT IN AEGILOPS LONGISSIMA By HONGYAN SHENG i A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy WASHINGTON STATE UNIVERSITY Department of Plant Pathology May 2011 © Copyright by HONGYAN SHENG, 2011 All Rights Reserved To the Faculty of Washington State University: The members of the Committee appointed to examine the dissertation of HONGYAN SHENG find it satisfactory and recommend that it be accepted. _________________________________________ Timothy D. Murray, Ph. D., Chair _________________________________________ Xianming Chen, Ph. D. _________________________________________ Scot H. Hulbert, Ph. D. _________________________________________ Tobin L. Peever, Ph. D. _________________________________________ Stephen S. Jones, Ph. D. ii ACKNOWLEDGMENT I would like to express my sincere gratitude and appreciation to my mentor and major advisor, Dr. Timothy D. Murray, for all his guidance, support, patience, and encouragement throughout my entire Ph. D. process at Washington State University. I am grateful to Dr. Murray for sharing his knowledge of plant pathology, providing insight into this dissertation, and leading me to the complex and fascinating world of genetics. My grateful appreciation goes to my committee members, Dr. Tobin L. Peever, Dr. Xianming Chen, Dr. Scot H. Hulbert, and Dr. Stephen S. Jones for their helpful advice and guidance during my graduate work and critical review of my dissertation. I would especially like to thank Dr. Deven R. See (USDA-ARS Regional Small Grains Genotyping Laboratory at Pullman, WA) for providing techniques and equipments for marker analysis work. Most of all, I am grateful for his critical suggestion leading to successful results. From all of these individuals, I have gained the knowledge and experience to be a successful scientist. I would also like to thank the Washington Grain Commission for financial support of my Ph.D. research, the USDA National Small Grains Collection for providing seeds of Ae. longissima accessions, and the Wheat Genetics and Genomic Resources Center at Kansas State University for providing seeds of Ae. longissima addition or substitution lines. I am also grateful to all members of Dr. Murray‟s and Dr. See‟s labs. Thanks to Dr. Kathy Klos and Dr. Henry Wetzel for their help and support in the lab and greenhouse, and to the students who assisted in this work. Special thanks to Dan iii Dreesmann for his support in the Plant Growth Facility Greenhouse. My appreciation also goes to all of the faculty, staff, and graduate students in the Department of Plant Pathology who provided help, encouragement, and friendship. I would especially like to thank Dr. Hanu Pappu, Dr. Brenda Schroeder, Cheryl Hagelganz, Mary Stormo, and Robin Stratton. My deep gratitude goes to my family back in Changchun, China. I greatly appreciate my father and mother for their sacrifices, encouragement, and understanding during my studies far from home. I also am grateful to my brother, sister, and all of my relatives and friends who take care of my parents. Finally, I would like to express my appreciation and love to my family here in the US. Thanks to my husband, Bo Gao, for walking with me through this long journey. Thanks also to my wonderful children, Lucy and Luke, for inspiring me all the time and understanding the time that went into this work when they grow up. iv IDENTIFICATION AND MAPPING OF RESISTANCE GENES FOR EYESPOT OF WHEAT IN AEGILOPS LONGISSIMA Abstract by Hongyan Sheng, Ph.D. Washington State University May 2011 Chair: Timothy D. Murray Eyespot of wheat, caused by two fungal pathogens Oculimacula yallundae and O. acuformis, produces elliptical lesions at the stem base that result in lodging of infected plants and yield losses up to 50% in commercial wheat fields. The objectives of this research were: 1) to identify potential new sources of resistance to O. yallundae and O. acuformis from a wild relative of wheat, Ae. longissima (2n = 14, SlSl); and 2) to understand the genetic control of the resistance and map gene(s) in the Sl genome by developing a genetic linkage map based on wheat microsatellite markers. Forty Ae. longissima accessions and 83 Ae. longissima addition or substitution lines in Chinese Spring or Selkirk background were evaluated for resistance to eyespot. Forty-three percent of the accessions were resistant to O. yallundae, 48% were resistant to O. acuformis, and 33% were resistant to both pathogens. This is the first evidence that Ae. longissima contains resistance to eyespot. Ae. longissima chromosomes 1Sl, 2Sl, 5Sl, and 7Sl contributed to the resistance for both O. yallundae and O. acuformis, and indicated that eyespot resistance was controlled by multiple regions of the Sl genome. Differential resistance reactions to O. yallundae and O. acuformis were observed in 25% of Ae. longissima accessions and 39% addition or substitution lines. v A recombinant inbred line (RIL) population was developed from the cross of Ae. longissima accessions PI 542196 (R) x PI 330486 (S) to map eyespot resistance genes. A genetic linkage map of the Sl genome was constructed with 169 wheat microsatellite markers covering 1261.3 cM in 7 groups. F5 lines (189) were tested for reaction to O. yallundae and four QTL were detected in chromosomes 1Sl, 3Sl, 5Sl, and 7Sl. These QTL explained 44% of the total phenotypic variation by GUS scores and 63% by visual ratings. This is the first time multiple QTL conferring resistance to eyespot in the Sl genome have been identified. Markers Xcfd6, Xwmc597, Xwmc415, and Xcfd2 are tightly linked to Q.Pch.wsu-1Sl, Q.Pch.wsu-3Sl, Q.Pch.wsu-5Sl, and Q.Pch.wsu-7Sl, respectively. The identification of these markers will facilitate QTL transfer to wheat by marker- assisted selection and broaden the genetic diversity of eyespot resistance. vi TABLE OF CONTENTS Page ACKNOWLEDGEMENTS ............................................................................................... iii ABSTRACT .........................................................................................................................v LIST OF TABLES ............................................................................................................. ix LIST OF FIGURES ............................................................................................................x CHAPTER 1. INTRODUCTION AND LITERATURE REVIEW .............................1 Wheat Production ........................................................................................1 Eyespot .........................................................................................................3 Host Resistance to Eyespot ........................................................................31 Objectives ..................................................................................................44 References ..................................................................................................45 2. IDENTIFYING NEW SOURCES OF RESISTANCE FOR EYESPOT OF WHEAT IN AEGILOPS LONGISSIMA .......................................64 Introduction ...............................................................................................64 Materials and Methods ..............................................................................67 Results .......................................................................................................72 Discussion .................................................................................................85 References .................................................................................................93 3. GENETIC ANALYSIS AND QTL MAPPING OF RESISTANCE GENES TO EYESPOT OF WHEAT IN AEGILOPS LONGISSIMA .....................................................................................98 Introduction ...............................................................................................98 vii Materials and Methods ............................................................................102 Results .....................................................................................................108 Discussion ...............................................................................................127 References ...............................................................................................137 4. INTERPRETIVE SUMMARY ..........................................................143 Future work .............................................................................................146 References ...............................................................................................148 viii LIST OF TABLES Page CHAPTER 1 Table 1. Virulence of eyespot isolates to seedlings of five host species (modified from Scott & Hollins, 1980) ..................................................................................11 CHAPTER 2 Table 1. GUS scores and disease ratings of 40 Aegilops longissima accessions to Oculimacula yallundae and O. acuformis .............................................................74 Table 2. GUS scores and disease ratings of 83 Ae. longissima addition or substitution lines in Chinese Spring or Selkirk background to Oculimacula yallundae and O. acuformis ..................................................................................80 CHAPTER 3 Table 1. Mean and standard deviation for GUS scores and visual ratings in three generations
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