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Half-Sib Selection for Higher Betalains Concentration and Lower Total Dissolved Solids in Table Beets (Beta Vulgaris)

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Half-Sib Selection for Higher Betalains Concentration and Lower Total Dissolved Solids in Table Beets (Beta Vulgaris) AN ABSTRACT OF THE THESIS OF Monzarath Hernandez for the degree of Master of Science in Horticulture presented on May 29, 2020. Title: Half-Sib Selection for Higher Betalain Concentration and Lower Total Dissolved Solids in Table Beets (Beta vulgaris). Abstract approved: ______________________________________________________ James R. Myers Betalains are a group of compounds that are major natural food colorants used by the food processing industry. These secondary compounds are found in only a few orders of plants with the Caryophyllales being the source of several domesticated crops. In particular, the family Chenopodiaceae in general and table beets (Beta vulgaris) specifically are the primary source for betalains for commercial extraction. Table beets are preferred because of high pigment concentration in the enlarged root in a crop that is relatively easy to grow, harvest and process. The primary betalains found in table beet are betacyanins (red pigments) and betaxanthins (yellow pigments). The food colorant industry is mainly interested in the betacyanin betanin, but betanin content is highly correlated with betalains so that selection for total betalains will result in an increase in betanin. Beets are also an economic source of sugars (primarily sucrose), which resulted in the development of sugar beets that are unpigmented but have sugar content of more than 20%. Table beets with moderately high sugar content have better flavor for culinary processes, but high sugars reduce efficiency of the extraction process of betalains for food colorants. Sucrose content in table beet is highly correlated with total dissolved solids (TDS), which can be easily measured with a refractometer whereas quantification of sucrose is more involved. Table beets with high betanin and low TDS are being sought by the food 2 concentrate industry. The objective of this study was to breed for high betalain pigment and low TDS in table beets. While quantification of betalains can be performed by UV-Vis spectrophotometry, the process is laborious and not suited to evaluating large numbers typically found in a breeding program. As a substitute for direct measurement of betalains, selection based on colorimeter measurements using the CIE L*a*b* color coordinate system was investigated. Parents were obtained from the USDA National Plant Germplasm System and from commercial sources. Twelve plant introduction accessions were chosen based on passport data in the GRIN database that indicated that they were pigmented and possessed low (<5%) sucrose content. Also selected were commercial lines with strong and uniform pigmentation, but moderately high TDS content. Six rounds of half-sib mass selection were performed to select simultaneously for increased pigment and decreased TDS. The process involved growing plants in the field in a late summer – early fall trial, evaluating pigment and TDS content of 10 roots per family and placing selected roots into cold storage (2°C) where they were vernalized for a minimum of 11-12 weeks. Roots were then transferred to the greenhouse in January and allowed to bolt, flower and random mate. Seeds were harvested in May and June and used for the next cycle field trial. From the accessions and cultivars grown in the field in 2014 - 2019, samples of 10 roots from each accession were taken for pigment and TDS analysis. From 20 - 30% of the individuals with high pigment and low TDS were retained in each cycle. In 2019, remnant seed from previous cycles along with the current cycle (cycle 5) were included in a replicated field trial so that all cycles could be evaluated in the same environment and heritability and gain from selection could be calculated. In conjunction with the colorimeter measurements, betalains were quantified in 2019 using UV-Vis spectrophotometer, and correlations between colorimeter and spectrophotometer data were calculated. For betalains, family means for CIE L* had a steady decease throughout the years indicating selection for lower L* resulted in families with darker pigmentation. TDS family averages, on the other hand, showed essentially no change over cycles. When reviewing data across years, two factors that may have influenced TDS levels were year to year environmental variation and the fact that betalains are usually present as a glycoside such that any increase in the amount of betalains will be associated with additional glyosidic content. Colorimeter measurements were converted to Royal Horticultural Society colors and RGB (Red-Green- Blue) color values. Over time, the half-sib population showed an increase in the purple-red and grey-purple groups. The population had a high narrow-sense heritability of 0.74, and also a relatively high change from selection of 0.75 per cycle for L*. TDS had low h2 and change from selection of 0.16 and 0.062% per cycle, respectively. In the case of the correlation of UV- VIS spectrophotometer data with colorimeter measurements, evaluation of six lines in the 2019 population showed a strong negative relationship between colorimeter and spectrophotometer values. However, there was a weak regression coefficient (r2=-0.03) for L* vs. betanin among families of the entire 2019 population. Overall, selection based on L* resulted in transgressive segregation for betanin, with some families outperforming commercial cultivar. Thus, the colorimeter appears to have utility as a technique for rapid and high throughput betalain measurement. ©Copyright by Monzarath Hernandez May 29, 2020 All Rights Reserved Half-Sib Selection for Higher Betalains Concentration and Lower Total Dissolved Solids in Table Beets (Beta vulgaris). by Monzarath Hernandez A THESIS submitted to Oregon State University in partial fulfillment of the requirements for the degree of Master of Science Presented May 29, 2020 Commencement June 2020 Master of Science thesis of Monzarath Hernandez presented on May 29, 2020 APPROVED: Major Professor, representing Horticulture Head of the Department of Horticulture 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. Monzarath Hernandez, Author ACKNOWLEDGEMENTS There is quite a large number of number of people who I am greatly appreciated for their assistance in the completion of my project. I owe my sincere appreciation to my principal investigator, Dr. Jim Myers for his creative insights, knowledge in the profession, and support during the grad school experience. I would like to show my gratitude Dr. Ryan Contreras, Dr. Yanyun Zhao for their guidance and assistance as thesis committee members. As well, Dr. Jean Hall was very sweet and kind to serve as a graduate council representative. Great thanks to Dr. Jennifer Kling with her assistance with data analysis for my research. Special thanks to Joel Davis and Shinji Kawai for their support and assistance in my project. To all my lab mates, grad and undergrad students throughout my grad school experience thank you for being a helping hand out in the field or greenhouse, and person to lean on when school would become overwhelming. To my parents, sisters and friends, thank you for encouraging and supporting my decision to go back to graduate school to pursue my dreams. In Deep Gratitude to Kerr Concentrates Inc. for funding this project. It was an honor to work on this project. Eugene Heuberger and Jose Guerrero, it was privilege working with you and learning from you about the food processing process for quantifying betalains from beet juice. TABLE OF CONTENTS Chapter 1 ...................................................................................................................................... 1 Introduction .............................................................................................................................. 1 Literature Review......................................................................................................................... 4 Introduction .............................................................................................................................. 4 Table Beets............................................................................................................................... 4 TDS: Comparison of Sugar Beets and Table Beets ................................................................. 8 Genes controlling Color ....................................................................................................... 9 Colorants, and their Market Value ......................................................................................... 10 Colorimetry ............................................................................................................................ 12 Objectives .............................................................................................................................. 14 Hypothesis.............................................................................................................................. 14 Chapter 2: Selection for Lower L* Value and Lower Sugar Concentration in Table Beets ..... 15 Introduction ............................................................................................................................ 15 Methods and Materials ........................................................................................................... 15 Production environments ..................................................................................................
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