Production of Native Plants for Seed, Biomass, and Natural Products A
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Production of native plants for seed, biomass, and natural products A Dissertation SUBMITTED TO THE FACULTY OF UNIVERSITY OF MINNESOTA BY Katrina Franziska Freund Saxhaug IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY Craig C. Sheaffer, advisor March 2020 © Katrina Franziska Freund Saxhaug 2019 Acknowledgements The research presented in this document would not have been possible without the love and support of countless mentors, colleague, friends and family. Foremost, I am forever grateful to my advisor, Dr. Craig Sheaffer, whose direction, support, understanding, and unending generosity made it possible for me to complete my doctorate. I am also eternally thankful for my unofficial co-advisor, Dr. Adrian Hegeman, for his kindness, intellectual brilliance, and support throughout my degree program. I am also incredibly grateful to Dr. Susan Galatowitsch and Dr. Clay Carter for their guidance, wisdom, and constructive and insightful commentaries. Though not on my committee, Dr. Jacob Jungers was incredibly generous with his time, advice, and support in all aspects of this research. While a doctoral student, I was supported by grants from the Minnesota Department of Agriculture through the AGRI Crop Research Grant Program and the Specialty Crop Block Grant Program. Additional support came through the Minnesota Institute for Sustainable Agriculture gift fund, graciously provided by Leanna Forcier. Further support was provided by the University of Minnesota, including the Hueg-Harrison Graduate Fellowship, the Mark and Jean Schroepfer Fellowship, the Nancy Jo Ehlke Fellowship, and Annie’s Sustainable Agriculture Scholarship. The Sustainable Cropping Systems Lab, under the direction of Dr Craig Sheaffer, and the Plant Metabolomics Lab, under the direction of Dr. Adrian Hegeman, were constant sources of support. I thank Joshua Larson, Brett Heim, Lindsay Wilson, and Katherine Bohn for their invaluable assistance with field work and the processing of field samples. I thank Dr. Dana Freund for her extensive guidance in the laboratory and for her moral support throughout my entire program. I thank Dr. Arthur Eschenlauer for his unceasing patience and the seemingly endless hours he spent training me to process large metabolomics datasets. I could not have completed my dissertation without the love and support of friends and family members, too numerous to list all here. I thank Courtney Jones for her entertaining conversations about “all things science”, and also for her encouragement to take time to just sit back and relax. I thank Katherine Sammons for her stimulating conversations and practical perspective. I am so very grateful to my parents, Thomas and Kay Freund, for encouraging my endless questions as a child and for putting an emphasis on education and exploration from an incredibly young age. I thank my sister and brother-in-law, Anna and Rick Peterson, for their nonstop support and reassurance. I am forever grateful for my husband, Mark Saxhaug, for his unwavering support throughout this entire program. I thank Mark for his assistance with field work on the hottest summer Saturdays and mundane laboratory tasks in the evenings. This would not have been possible without his love, understanding, support, and extraordinary sense of humor that could lighten any mood. Finally, I am thankful for several relatives who, although they have not been with me several years, still inspire me every day. My grandfather, John William Freund, who was i my first instructor of the natural world and taught me the names of wildflowers, trees, and animals. My Great Uncle Raymond Kabot, who loved farming but also took the time to wander the prairies of southwest Minnesota to find flowers. My Great Aunt Barbara Teckla Kabot, who taught me gentleness, kindness, and how to manage a farm and home through love and compassion. Finally, my Great Aunt Frances Edna Kabot for her hard work ethic, adventurous spirit, support of education, and whose love of the outdoors lead to endless hours lost in the garden. I dedicate this dissertation John, Raymond, Barbara, and Frances. ii Abstract Native and naturalized perennial plants are important components of sustainable agricultural systems, providing a wide range of ecosystem services including marketable products. Although demand for these products is increasing, there is limited information on the establishment and production of native and naturalized perennial plants. Through field and greenhouse studies, four native and naturalized perennial plant species were evaluated for their seed yield, biomass yield, and phytochemical (natural product) production. Canada milk vetch (Astragalus canadensis L.), purple coneflower (Echinacea purpurea Moench [L.]), and showy tick trefoil (Desmodium canadense L.) were evaluated in the field for seed and biomass production over three consecutive years. Seed yield decreased in all three species over time, whereas biomass yields over time varied by species. Seed and biomass yields, on a per hectare basis, decreased as the number of rows decreased, and there was little effect of plant community richness or diversity. An untargeted metabolomics approach was taken to evaluate phytochemical production in purple coneflower tissues. Several bioactive compounds were provisionally identified in purple coneflower tissues, and seed tissue possessed a similar profile to that of root. Although the plant community did not affect the overall metabolomic profile of purple coneflower, levels of specific compounds in leaf, stem, and root tissue were affected. Similar biomass yield and metabolomics approaches were applied to fireweed (Chamaenerion angustifolium (L.) scop., syn. Epilobium angustifolium L.) grown in a hydroponic system. Shoot dry weight increased logistically with increasing mineral concentration of the nutrient solution. Although the abundance of most provisionally identified compounds decreased with increasing mineral nutrient concentration, iii commercially-relevant oenothein B and miquelianin were not affected by mineral nutrient treatments. Native and naturalized perennial plants on the agricultural landscape can provide numerous ecosystem services, including marketable products such as seed for restoration plantings, biomass for fuel and forage, and bioactive phytochemicals for medicinal and supplemental purposes. In order to incentivize the establishment of native and naturalized perennials for such marketable products, research on the productivity of specific plants is needed. The research presented in this dissertation provides the groundwork for four native and naturalized Minnesota species. iv Table of Contents Acknowledgements……………………………………………………………………….. i Abstract………………………………………………………………………………….. iii Table of Contents……………………………………………………………………….... v List of Tables………………………………………………………………………….… vi List of Figures……………………………………………………….…………………. viii Introduction………………………………………………………………………………. 1 Chapter 1: Cultivation of native plants for seed and biomass yield……………………... 6 Chapter 2: Field production of purple coneflower (Echinacea purpurea Moench [L.]) for beneficial phytochemicals………………………………………………………………. 35 Chapter 3: Hydroponic production of fireweed (Chamaenerion angustifolium (L.) scop.) for biomass and phytochemical production…………………………………………..… 69 Conclusion…………………………………………………………………………….. 105 Bibliography…………………………………………………………………………... 109 Appendix A. Chapter 1 Supporting Information……………………………………… 134 Appendix B. Chapter 2 Supporting Information………………………………….…… 139 Appendix C. Chapter 3 Supporting Information…………………………………….… 145 v List of Tables Table 1.1. Average mean daily temperature (°C) and mean monthly total precipitation (mm) from 2016 to 2018. ------------------------------------------------------------------------ 28 Table 1.2. Additional species included in the low-richness and high-richness polyculture plots. ------------------------------------------------------------------------------------------------- 29 Table 1.3. Mean seed, focal biomass, and total biomass yield for main effects and interactions for the three focal species in Becker and Rosemount. ------------------------- 30 Table 2.1. Tissue-specific details on the metabolomics harvest of purple coneflower flower, leaf, stem, root, and seed. --------------------------------------------------------------- 59 Table 2.2. Features provisionally identified in purple coneflower tissues by UHPLC– Orbitrap-MS. --------------------------------------------------------------------------------------- 60 Table 3.1. Stock solution requirements for Hoagland nutrient solutions. ----------------- 96 Table 3.2. Recipes for the six different Hoagland nutrient solutions. --------------------- 97 Table 3.3. Tissue-specific metabolomics harvest details for hydroponic fireweed. ----- 97 Table 3.4. Provisionally identified features in hydroponically-cultivated fireweed. ---- 98 Supplementary Table A.1. Seed and biomass harvest dates for the three focal species at two locations. ------------------------------------------------------------------------------------- 134 Supplementary Table A.2. Mean focal seed yield (kg ha-1) of the six designs and three focal species in Becker and Rosemount 2016-2018. ---------------------------------------- 135 Supplementary Table A.3. Mean focal biomass yield (kg ha-1) of the six designs and three focal species in Becker and Rosemount 2016-2018. --------------------------------- 136 Supplementary Table A.4. Mean total biomass yield (kg ha-1)