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Awalemani.Pdf CHARACTERIZATION OF BERRY AND WINE AROMA DEVELOPMENT IN INTERSPECIFIC HYBRIDS IMPORTANT TO MISSOURI USING A METABOLOMICS BASED APPROACH _______________________________________ A Dissertation presented to the Faculty of the Graduate School at the University of Missouri-Columbia _______________________________________________________ In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy _____________________________________________________ by MANI AWALE Dr. Misha Kwasniewski, Dissertation Supervisor JULY 2020 The undersigned, appointed by the dean of the Graduate School, have examined the dissertation entitled CHARACTERIZATION OF BERRY AND WINE AROMA DEVELOPMENT IN INTERSPECIFIC HYBRIDS IMPORTANT TO MISSOURI USING A METABOLOMICS BASED APPROACH presented by Mani Awale, a candidate for the degree of doctor of philosophy, and hereby certify that, in their opinion, it is worthy of acceptance. Professor Misha Kwasniewski Professor Walter Gassmann Professor Laszlo Kovacs Professor Ruthie Angelovici Dedicated to my late father who has always encouraged me to pursue my dreams, to my mom who has helped me navigate my path to my dreams and to my late grandmother who had always been my pillar of strength. Thanks, Mom. ACKNOWLEDGMENTS I am extremely grateful to the number of faculties, friends, and my family for all the encouragement, guidance, love, and support given to me throughout this adventurous, joyful as well as challenging journey. First, I express my profound gratitude to my advisor Dr. Misha Kwasniewski for providing me the precious opportunity to work on several interesting projects at the University of Missouri-Columbia. I am indebted to him for all his guidance, love, care, and immense knowledge throughout my research period. I still remember it was a challenging transition for me to join the lab, but I was excited. Before joining the lab, my background was in Plant (Grapes) Genetics. During my MS, I got a few opportunities to visit the local vineyards and winery and learned about the winemaking process, which was a fascinating moment. This reminded me of how my mom and grandma used to make Nepalese traditional wine and liquor back at home and got me more excited. This intensified my interest in the field of viticulture and enology after realizing that knowledge of genetics along with various management practices and winemaking processes can be an asset to improve the wine industry. I did not know how challenging the analytical chemistry will be for me until I started doing it myself. Thank you, Misha, for providing me an unquantifiable amount of guidance and support especially on chromatography and analytical chemistry without which I would not be able to succeed. I also thank Drs. Walter Gassmann, Ruthie Angelovici, and Lazlo Kovacs for serving in my committee and for providing me with constructive and meaningful comments and suggestions. Thank you very much for writing several recommendation letters on my behalf to different scholarship committees and organizations. I am truly indebted for that. My deepest gratitude goes to the Department of Plant Science, faculty ii and staff for all their help and support. I am very much thankful to Grape and Wine Institute and NSF VitisRoots for funding my research. Thank you to the entire VitisRoots team especially to Dr. Allison Miller and Dr. Anne Fennell (my former MS advisor, SDSU) for being my role model for Women in Science and all the help and support. My sincere thanks to Connie Liu for her contribution and assistance in running my samples and for her love and care in the lab. Thank you, Connie, for being both my mentor as well as my friend. Your contribution to running samples and troubleshooting has helped me to move forward. I cannot forget my mentor and friend Megan for her useful suggestions, motivation, and hearing to me when I needed someone. Thank you for providing me the positive vibes and the right attitudes all the time. I am thankful to all my friends and lab colleagues for their love and cooperation. Shout out to Alex, Alan, Joe, Atzin, and lovely Maggie for all the help and fun. I appreciate the love and support given to me by my family, my mom, without which I couldn't stand at this stage. She has been an example of courage and strength to me. Although she can barely read and write, her love, sacrifice and continued support for my education even after the passing of my Dad at an early age, has motived me to pursue this dream, and to be determined to make this dream come true. My grandma and my aunt also have had my back throughout my life. I thank them and my brother, Abin, and my sister in law, Mimi for always being with me. My interest in the field of agriculture developed from our family farms where we used to plant rice in summer. Thank you for teaching me to plant rice and instilling me that hard work and patience is required to bear fruit. I also owe this to my childhood best friends Sabina and Sareena who always cheered me throughout my difficult times. I also thank my in-laws especially Vivek’s iii sisters who had been the sisters I always wanted and who had been so welcoming and supportive to me throughout this time. Lastly, my dearest love and appreciation goes to my husband, Vivek, for the motivation, encouragement, love, and support and for always being with me in my sorrow and happiness. Thank you for listening to those countless problems and always motivating and supporting me when I was ready to give up. Thank you for believing in me when I doubted myself. To anybody that I failed to mention, I thank you all so much for your help. iv TABLE OF CONTENTS ACKNOWLEDGMENTS .................................................................................................. ii LIST OF FIGURES ............................................................................................................ x LIST OF TABLES ............................................................................................................ xv ABSTRACT:.................................................................................................................... xvi Chapter 1: General Introduction and literature review ....................................................... 1 1. Fruit and wine chemistry: ............................................................................................ 2 1.1 Organic acids: ....................................................................................................... 4 1.2 Flavor profile of grapes ........................................................................................ 5 1.3 Free and bound volatiles in grapes ....................................................................... 6 1.4 Grape aroma molecules: ....................................................................................... 8 Terpenes: ..................................................................................................................... 8 Norisoprenoids: ......................................................................................................... 11 Esters: ........................................................................................................................ 11 Aliphatic alcohols: ..................................................................................................... 12 Aliphatic acids: .......................................................................................................... 12 2. Hybrid grapes and their aroma .................................................................................. 13 2.1 Norton grapes and wines: ................................................................................... 14 2.2 Chambourcin grapes and wines.......................................................................... 15 3. Plant Metabolomics ................................................................................................... 16 3.1 Targeted and untargeted metabolomics................................................................... 17 3.2 Untargeted Metabolomics Workflow ...................................................................... 18 3.3 Solid-phase microextraction (HS-SPME): .............................................................. 19 3.4 Analytical Methodology:......................................................................................... 19 3.5 Data pre-processing and pre-treatment: .................................................................. 21 3.5 Statistical analysis: .................................................................................................. 23 3.6 Multivariate analysis: .............................................................................................. 23 3.7 Univariate Analysis ................................................................................................. 24 4 Development and use of rootstocks in viticulture: .................................................... 26 5 Rootstocks effect on grapes and wine chemistry: ..................................................... 27 v 6 Irrigation effect on grapes and wine chemistry: ........................................................ 29 7 Midday Stem Water Potential: .................................................................................. 31 8 References ................................................................................................................. 32 Chapter 2: Identification of aroma markers for Norton and Cabernet Sauvignon berries and wines to characterize
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