AN ABSTRACT OF THE THESIS OF Karli R. Van Simaeys for the degree of Master of Science in Food Science and Technology presented on December 15, 2020. Title: Examining Environmental and Genetic Influences on the Brewing Performance of Hops (Humulus lupulus) and Barley (Horduem vulgare). Abstract approved: ______________________________________________________ Thomas H. Shellhammer In short, the brewing process uses hot water to extract fermentable sugars from malt to make a wort that is bittered by hops and finally fermented by yeast to produce beer. The four key ingredients in brewing are malt, water, hops, and yeast. Malt is perhaps the key ingredient, as it contains starches and protein as well as the enzymes required to break them down and is the source of fermentable extract that is ultimately converted to beer during an alcoholic fermentation. Hops, the cones of the female Humulus lupulus plant, are used for both bittering beer and providing aromatic qualities. Hops are grown in various regions globally and differences in soil, weather, climate, disease pressure, and grower practices provide inherent variation among them. This variation in hop growing conditions leads to differences in final hop quality, which growers have begun to focus on in recent years. This relationship, called terroir or regional identity, examines how the quality of agricultural products relate to their place of origin. It encompasses the soil, weather and climate, topography of the growing location, and to a certain degree the influences of grower management in response to these influences. Specified growing regions have been established for other agricultural products, such as wine grapes, coffee, cocoa, and tea. Anecdotal knowledge and published research show early evidence of regional identity affecting hops. Several studies have shown differences in hop chemistry and sensory qualities among samples grown in different locations. However, more work is needed to understand the extent of regional variation and to determine which factors drives differences among hops grown in different locations. To assess the potential effect of regional identity on hops, three varieties were harvested from a total of eleven commercial fields managed by a single hop grower within the Oregon Willamette Valley. Weather, climate, management practices, and soil data were collected for each site. Multiple Factor Analysis of these data sets showed evidence of a location effect. As expected, chemical analysis showed sample groupings by variety but variation within each variety demonstrated the effect of growing location. Sensory evaluation showed significant differences between samples within the same variety. Variation within field and between fields of the same variety provide early evidence of regional influences on hop qualities. Seeds from the barley plant, Hordeum vulgare, are unsuitable for brewing until they are transformed via controlled germination and kilning during the malting process. While malting is a significant driver of malt flavor, recent work shows that other factors, such as genetics, may also influence flavor. To evaluate the contributions of barley genotype to beer flavor, two independent sets of barley germplasm were evaluated. Pedigree, quality of malt and beer, and beer metabolomic profiles were compared within and between the two sets. Sensory attributes of malt hot steeps and lager beers were evaluated, and distinct but subtle differences were reported. Distinct metabolomic profiles, attributable to barley genotype, were detected. In conclusion, metabolite variation observed is a direct result of genetic differences that lead to differential chemical responses within the malting and brewing processes, thus affecting flavor. ©Copyright by Karli R. Van Simaeys December 15, 2020 All Rights Reserved Examining Environmental and Genetic Influences on the Brewing Performance of Hops (Humulus lupulus) and Barley (Horduem vulgare) by Karli R. Van Simaeys A THESIS submitted to Oregon State University in partial fulfillment of the requirements for the degree of Master of Science Presented December 15, 2020 Commencement June 2021 Master of Science thesis of Karli R. Van Simaeys presented on December 15, 2020 APPROVED: Major Professor, representing Food Science and Technology Head of the Department of Food Science and Technology Dean of the Graduate School I understand that my thesis will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my thesis to any reader upon request. Karli R. Van Simaeys, Author ACKNOWLEDGEMENTS I would like to begin by expressing my gratitude to Dr. Tom Shellhammer for the opportunity to be a part of the lab for the last two years. Your mentorship and depth of knowledge were invaluable during my time as a Master’s student. With your enthusiasm and guidance our lab has become a team that can achieve a great deal together. To my lab mates, past and present, I could not have done this without your support and friendship. I am looking forward to seeing what you all go on to accomplish. I would also like to thank Jeff Clawson for his expertise and dedication to the brewing projects I have endeavored on. I am sincerely grateful you were able to share your brewing knowledge with me and for your patience in that process. I also would like to thank Dr. Elizabeth Tomasino for your guidance on sensory data collection and analysis throughout several projects. Special thanks go to Dr. Pat Hayes and his lab, for welcoming me into Barley World and giving me the opportunity to learn about barley and malt. Additionally, I want to thank Coleman Agriculture for their partnership and hop expertise, particularly Liz Coleman and Garrett Weaver. I am also grateful for my time during my internship at Anheuser-Busch, and for Cindy-Lou Lakenburges, Adriana Lezama, and Weilun Tsai in particular. Finally, I want to say how grateful I am for my family and friends. I would not be here without your encouragement and support. Thank you for everything. CONTRIBUTION OF AUTHORS For Chapter 2, several authors contributed to the project. My contributions included gathering data, executing sensory analysis, performing all statistical analysis and interpretation. Andy Gallagher performed soil mapping, soil coring, and interpretation. Garrett Weaver assisted with site selection, gathering farm management data, and project guidance. Arnbjørn Stokholm assisted with harvest and performed hop chemistry analysis. Dr. Tom Shellhammer conceived the project concept and identified research goals, acquired funding, contributed to project planning, data interpretation and manuscript reviewing and editing. For Chapter 3, the work was divided into three sections, malt production and genetics, brewing and sensory, and metabolomics. My contributions to the project were executing laboratory panel sensory testing and analyzing and interpreting the results. My contributions will be included in Chapter 3, while the full publication will be attached as an appendix. Sarah Windes, Scott Fisk, and Dr. Patrick Hayes developed the experimental varieties and were responsible for malting. Harmonie Bettenhausen and Dr. Adam Heuberger performed metabolomics and analysis. Jeff Clawson brewed the experimental beers. Dr. Juyun Lim and Sue Queisser contributed the consumer sensory data. Dr. Tom Shellhammer provided guidance with data interpretation and contributed to manuscript review and editing. TABLE OF CONTENTS Page Chapter 1 – General Introduction ...……………………………………………...……… 1 Brewing Process ………..………...……………………………………....…1 From Barley to Malt ………………...…….……………………………...…3 Hops ………………...……….………...…………………….……………...5 Regional Identity……………..………...…………………....………………8 References ……………...……………...…………………………………..11 Chapter 2 – Examining regional differences of new American aroma hops grown in the Willamette Valley, Oregon…………………………….……….………………... 14 Abstract …...………………………………………………………….……14 Introduction ……….………………………………………………………14 Materials and Methods ……………………………………………………18 Results and Discussion …….……………………………………………...30 Conclusion…………………………………………………………………45 Acknowledgements…..…………………………………………………….46 References …….………...……………...………………………………….48 Supplementary Material…..…………………………….…………………..52 Chapter 3 – Excerpt from Comprehensive Analysis of Different Contemporary Barley Genotypes Enhances and Expands the Scope of Barley Contributions to Beer Flavor….……………………………………………………………………..…..…58 Introduction ……….………………………………………………………59 Sensory Methodology……………………………………………………...59 TABLE OF CONTENTS (Continued) Results ……………….…….……………………………………………...61 Discussion …….…………………………………………………………...65 Conclusions…………………………………………...……………………69 References………………………………………………………………….71 Supplementary Material…..……………………………….…………….....72 Future Work.………………………………….……………………………………….....76 Bibliography…………………………..………………………………………….....…...78 Appendices……………………………………………………………………………….87 Full Publication - Comprehensive Analysis of Different Contemporary Barley Genotypes Enhances and Expands the Scope of Barley Contributions to Beer Flavor……………………………………………………………………...87 LIST OF FIGURES Figure Page Chapter 1 Figure 1: Summary of brewing process and points of ingredient addition……...……...…1 Chapter 2 Figure 1: Map of Oregon commercial hop growing region within the Willamette Valley………………………………………………………………………..……….…..16 Figure 2: Soil map and sampling sites for Alluvial 33 Mosaic® field (MOS – All 33)…………………………………………………………………..………………..21 Figure 3: Soil map and sampling