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Characterization and Identification of Novel Reduced Height (Rht-1)

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Characterization and Identification of Novel Reduced Height (Rht-1) CHARACTERIZATION AND IDENTIFICATION OF NOVEL REDUCED HEIGHT (RHT-1) ALLELES IN WHEAT by Emma Marie Jobson A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Plant Genetics MONTANA STATE UNIVERSITY Bozeman, Montana November 2019 ©COPYRIGHT by Emma Marie Jobson 2019 All Rights Reserved ii DEDICATION To my grandpa, Reuben Swanson, who taught me: the keys to success are faith, arable land, and a good education. To my parents, my brother, and Jake; thank you for your encouragement and support throughout this endeavor. iii ACKNOWLEDGEMENTS I would like to express my sincere appreciation to those who have helped me throughout my graduate school experience. Firstly, I would like to thank my advisor, Dr. Mike Giroux. His guidance, patience, and support has been constant throughout my time at Montana State University. He has assisted me through the entirety of my program and I am truly grateful for everything he has taught me. I would like to thank my committee members, Dr. Phil Bruckner, Dr. Robert Sharrock, and Dr. Andreas Fischer for their advice and assistance. Particularly, I would like to express my gratitude to Dr. John Martin for his support designing projects, analyzing data, and editing papers. I would also like to thank Dr. Martin for his continued willingness and enthusiasm as a teacher and his encouragement throughout my time at Montana State. I would like to thank my fellow lab members Rachel Johnston, Alanna Oiestad, Kendra Hertweck, Tavin Schneider, and Maddison Milliman for their assistance throughout my program. I am especially grateful for the help and patience of our lab manager Andy Hogg. I would also like to extend my gratitude to Deanna Nash and Harvey TeSlaa in the MSU Cereal Quality Lab for their assistance in conducting the end use quality testing. Finally, thank you to the current and former MSU PSPP graduate students who have provided feedback along the way including my current and former lab mates: Darby Kammeraad, Hannah Turner, Justin Vetch, and McKenna Brown. iv TABLE OF CONTENTS 1. INTRODUCTION ...........................................................................................................1 History and Significance of Rht-1 ....................................................................................2 Agronomic Impact of Rht-B1b and Rht-D1b ...................................................................2 Rht-1 encodes DELLA proteins ......................................................................................3 Literature Cited ................................................................................................................8 2. THE IMPACT OF THE WHEAT RHT-B1B SEMI-DWARFING ALLELE ON PHOTOSYNTHESIS AND SEED DEVELOPMENT UNDER FIELD CONDITIONS ...................................................................................12 Contribution of Authors and Co-Authors ......................................................................12 Manuscript Information Page .......................................................................................13 Abstract ..........................................................................................................................14 Introduction ....................................................................................................................15 Materials and Methods ...................................................................................................19 Plant Material .........................................................................................................19 Growing Conditions ...............................................................................................19 Plant Sampling .......................................................................................................20 Agronomic Measurements .....................................................................................22 Photosynthetic Measurements ...............................................................................23 Chlorophyll Measurements ....................................................................................24 Grain Weight, Number of Grains per Head, Grain Protein, and Starch Content .................................................................................................25 Leaf Protein and Starch Content ............................................................................26 Expression Analysis ...............................................................................................27 Leaf and Grain Metabolites ...................................................................................28 Statistical Analysis .................................................................................................28 Results ...........................................................................................................................29 Agronomic Measurements .....................................................................................29 Photosynthetic Capacity.........................................................................................29 Leaf Protein and Starch Content Throughout Photoperiod ....................................30 Starch and Protein Content of Grain Throughout Development ...........................30 Seed Number and Individual Kernel Weight Throughout Development ..........................................................................................................31 Effect of Rht-B1b Mutation on Metabolite Levels ................................................31 Transcript Analysis of Leaf and Stem mRNA .......................................................32 Discussion .....................................................................................................................32 Acknowledgements .......................................................................................................37 Literature Cited .............................................................................................................45 v TABLE OF CONTENTS CONTINUED 3. THE IMPACT OF THE RHT-B1B, RHT-D1B, AND RHT-8 WHEAT SEMI-DWARFING GENES ON FLOUR MILLING, BAKING, AND MICRONUTRIENTS ......................................................50 Contribution of Authors and Co-Authors ......................................................................50 Manuscript Information Page .......................................................................................51 Abstract ..........................................................................................................................52 Introduction ....................................................................................................................53 Materials and Methods ...................................................................................................56 Plant Material .........................................................................................................56 Field Design and Conditions ..................................................................................57 End Use Quality Analysis ......................................................................................57 Statistical Analysis .................................................................................................59 Results ............................................................................................................................59 Discussion ......................................................................................................................61 Conclusion .....................................................................................................................64 Acknowledgements ........................................................................................................65 Literature Cited ..............................................................................................................73 4. RHT-1 SEMI-DWARFING ALLELES ALTER DOUGH RHEOLOGY BY MODIFYING GLUTEN COMPOSITION ......................................78 Contribution of Authors and Co-Authors ......................................................................78 Manuscript Information Page .......................................................................................79 Abstract ..........................................................................................................................80 Introduction ....................................................................................................................81 Materials and Methods ...................................................................................................85 Plant Material .........................................................................................................85 Field Design and Conditions ..................................................................................85 Milling....................................................................................................................86 Flour and Dough Rheology ....................................................................................86 Starch Granule Visualization .................................................................................87 Protein Molecular Weight Distribution .................................................................87 RNA Sequencing ...................................................................................................89
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