WHEAT Ssrs PRIMERS SCREENING
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ABSTRACT WOOTEN, JR., DAVID RUDOLPH. New genetic tools for improving oat winter hardiness. (Under the direction of David P. Livingston, III and J. Paul Murphy). Winter hardiness is a complex trait, and poor winter hardiness limits commercial production of winter oat. The identification of new genetic tools to improve oat winter hardiness will allow for continued increases in oat winter hardiness. The objectives of this research were: identify quantitative trait loci (QTL) for crown freeze tolerance in the ‘Kanota’ x ‘Ogle’ mapping population, evaluate the winter hardiness component traits winter field survival, crown freeze tolerance, heading date, plant height, and vernalization responses in a population derived from a cross of winter tender ‘Fulghum’ with winter hardy ‘Norline’, examine the relationships between winter hardiness component traits and a reciprocal intergenomic translocation between chromosomes 7C and 17 in the Fulghum x Norline population, develop a genetic linkage map in the Fulghum x Norline population using SSR markers and selected RFLP markers, and identify QTL for winter hardiness component traits in the Fulghum x Norline population. In the Kanota x Ogle population seven QTL and four complimentary epistatic interactions were identified that accounted for 56% of the phenotypic variation. Ogle contributed alleles for increase crown freeze tolerance at three loci, while Kanota contributed alleles for increase crown freeze tolerance at four loci. All loci where Kanota alleles increased crown freeze tolerance showed complementary epistasis for decreased crown freeze tolerance with the QTL near UMN13. Two of the major QTL identified were in the linkage groups associated with the 7C-17 translocation. In the Fulghum x Norline population, 7C-17 translocation did not segregate in the expected 1:1 ratio, and twice as many translocation types as non-translocation types were observed. The translocation was significantly correlated with crown freeze tolerance (r=0.72) and winter field survival (r=0.62). The heritability of crown freeze tolerance was 83% and the heritability of winter field survival was 76%. Field heading date was significantly correlated with translocation status (r=0.20). Plant height, vernalization response, and photoperiod response were not associated with the translocation. QTL were identified for winter field survival, crown freeze tolerance, vernalization response, plant height and heading date, and epistatic interactions were identified for all of these traits except plant height. Major QTL for winter field survival (R2=35%) and crown freeze tolerance (R2 =52%) were identified on linkage group FN3 which was associated with the 7C-7 translocation. In this research, QTL for freeze tolerance in known linkage groups were identified for the first time in oat and specific QTL for winter hardiness component traits were identified in winter oat for first time. The importance of the 7C-17 translocation to oat winter hardiness traits was confirmed. Comparison with previous results explained differential performance between the hardy winter oat cultivars Norline and Wintok. These results suggested how the 7C-17 translocation conferred increased biological fitness in most Avena germplasm. NEW GENETIC TOOLS FOR IMPROVING OAT WINTER HARDINESS by DAVID RUDOLPH WOOTEN, JR. A dissertation submitted to the Graduate Faculty of North Carolina State University in partial fulfillment of the requirements for the Degree of Doctor of Philosophy CROP SCIENCE Raleigh, North Carolina 2006 APPROVED BY: _____________________________ _____________________________ David A. Dickey James B. Holland _____________________________ _____________________________ David P. Livingston, III J. Paul Murphy Co-chair of Advisory Committee Co-chair of Advisory Committee DEDICATION I dedicate this work to my grandparents Harry and Fran Rouse. One of my earliest memories is of planting watermelons on their farm in western Illinois. They started me on this path many years ago. I am very happy where it has led me. ii BIOGRAPHY David Wooten was born in West Columbia, South Carolina. He was raised in Colorado Springs, Colorado, until moving to Houston, Texas just before high school. He graduated from Klein High School and attended both the University of Texas at Austin, and Texas A&M, College Station before graduation from Texas A&M with B.S. degrees in Genetics and Plant and Environmental Soil Science in 1999. He continued his studies at Texas A&M working in the sorghum breeding program under the direction of Dr. William Rooney. In 2001 he received a M.S. degree in Plant Breeding and left Texas to begin studies at North Carolina State University. Beginning in 2002, he worked in the small grains breeding program at NC State. His dissertation research examined genetic tools for improving oat winter hardiness, under the direction of David Livingston and Paul Murphy. He finished his PhD in Crop Science in 2006 and accepted a position as a soybean breeder for Monsanto in Stonington, Illinois. iii ACKNOWLEDGEMENTS Firstly, I would like to thank my committee members for there encouragement, assistance, and patience in the completion of this research. Sometimes things did not go well, or I did not do well, but they stuck with me and gave me the opportunity and the help I needed to make my time at NCSU a great success. My colleagues in the small grains breeding program provided me with incomparable assistance in my work, and were such good friends to me. I will always remember mi Jefecito, René Navarro, and my fellow students, Dr. Goran Srnić, Dr. Lilian Miranda, Hong Mia Jia, and Judd Maxwell. Sometimes it was tough harvesting oats when it was in the upper 90’s or keeping our hands in ice water for hours at a time, but working together we made it. I had some hard times in the lab, but I made it through thanks to the help of Dr. Jennifer Tarter, Dr. Steve Szalma, David Rhyne, Josie Bloom, and Jeanette Lyerly. Thanks to them, I got the data that I needed without going completely crazy or contaminating all of Williams Hall with radioactive isotopes. Finally I must thank my family for their love and support through the years. That has always meant the most to me, and helped me when I needed it most. iv TABLE OF CONTENTS Page LIST OF TABLES....................................................................................................................... viii LIST OF FIGURES .......................................................................................................................x LITERATURE REVIEW ..............................................................................................................1 Introduction........................................................................................................................2 Oat usage and Production ..................................................................................................4 Origin.................................................................................................................................5 Cytogenetics ......................................................................................................................6 Winter Hardiness ...............................................................................................................7 Freeze Damage ............................................................................................................8 Intracellular Ice......................................................................................................8 Adhesion................................................................................................................9 Dehydration ..........................................................................................................9 Freeze Tolerance.........................................................................................................10 Freeze Testing.............................................................................................................13 Genetics of Winter Hardiness...........................................................................................15 Genetic Mapping ..............................................................................................................19 Quantitative Trait Loci .....................................................................................................21 References Cited...............................................................................................................24 Quantitative trait loci and epistasis for crown freeze tolerance in the Kanota x Ogle hexaploid oat mapping..................................................................................................................39 Abstract.............................................................................................................................40 Introduction.......................................................................................................................41 Materials and Methods .....................................................................................................43 Phenotypic Evaluation................................................................................................43 QTL Detection............................................................................................................45 Examination of Epistasis ............................................................................................46