COLD-CLIMATE GRAPE BREEDING AND ENOLOGICAL BEST PRACTICES

• Sponsoring Institution: State Agricultural Experiment Station
• Project Status: COMPLETE
• Funding Source: STATE
• Reporting Frequency: Annual
• Accession No.: 1009969
• Project Start Date: Jul 1, 2016
• Project End Date: Jun 30, 2021
• Program Code: [(N/A)]- (N/A)

Recipient Organization
UNIV OF MINNESOTA
(N/A)
ST PAUL,MN 55108

Performing Department
Horticultural Science

Non Technical Summary
Cold-hardy, hybrid grape cultivar development can be advanced through both traditional and molecular genetic approaches. The University of Minnesota (UMN) grape breeding project aims to develop products that spans several distinct sectors including wine grape; table grape; food, beverage, and food products; as well as ornamental and landscape plants. Advancements within the project include the development and testing of DNA markers for early screening of seedlings prior to field planting for important traits, and the use of DNA tests to select top quality parents for performing elite crosses. To date, we have marker tests for several traits including: disease resistance (several powdery mildew genes), flower sex, and seedlessness. This proposal aims to implement routine marker assisted breeding (MAB) as a tool to predict seedling performance and accelerate areas within the breeding timeline, especially for developing disease resistant varieties. Research in wine making will complement variety development, but will also focus on best practices for commercial wine production using cold-hardy grape cultivars. Research driven knowledge on wine making and grape growing will be disseminated through education and extension programming.

Animal Health Component

90%

Research Effort Categories

Basic

10%

Applied

90%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
201	1139	1081	100%

Knowledge Area
201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
1139 - Grapes, general/other;

Field Of Science
1081 - Breeding;

Keywords

genetics

breeding

grape

wine science

Goals / Objectives
Advance hybrid grape cultivar development for (a) cold-hardy wine grape production, (b) table grape markets, (c) food, beverage, and food product industry, and (d) ornamental landscape installations through traditional and high-throughput phenotyping methods.Implement routine marker assisted breeding (MAB) in grape cultivar development through (a) identifying marker-trait associations, (b) developing DNA tests for associated traits, (c) implementing DNA tests for screening potential parents and culling offspring.Develop protocols and best enology practices for commercial wine production using cold-hardy grape cultivars.Advance the understanding and practice of grape growing and wine making across the state through outreach and extension programming.

Project Methods
Cultivar development--Two general approaches are used for hybrid grape cultivar development, traditional breeding methods and marker assisted breeding (covered below). For over 150 years, traditional breeding methods have been applied as a way to develop new varieties, especially those with disease and pest resistance, and improved fruit quality traits for wine and table uses. Parents are selected for a cross based on their complementary traits of interest the breeder would like to combine in an offspring. A family is created, and seedlings are evaluated in the first year in a nursery for growth habit and disease resistance, then planted and evaluated in 3-5 years when the offspring will flower and fruit for the first time. The breeder then evaluates the population as a whole, and each individual within the cross. Approximately one of every 1000 vines each year is identified to become a parent or potential cultivar. The first culling decisions are based on visual performance of the vine and organoleptic properties of fruit quality by a single observer in the field.Juice chemistry analyses (total acidity, soluble solids, pH) are conducted for all samples whether for wine or table grape. Superior wine grape selections, those samples falling within the wine making standards, are also processed for microvinification. This process is repeated annually with new selections being either culled, selected, or evaluated for multiple years. The most elite selections are propagated to increase the number of plants available in order to evaluate different vineyard sites, test within multi-state projects, and to increase the number of fruit available for wine analysis. Sensory analysis with finished wine products is also important for determining the suitability for wine making. In addition to wine products, the breeding program is currently evaluating individuals for performance as table grapes, landscape plants, and food products (juice and dyes).Market research: The UMN grape breeding program has in its development a number of potential table grape cultivars that are an improvement over available cold-hardy varieties. Very little is known about consumer preferences for, market readiness, and the economic viability of new Minnesota-grown table grape cultivars. In order to have buy-in from nursery propagators and growers (a critical step during commercialization), market research is needed to investigate market potential and feasibility of this "new" horticultural product. Research will determine consumer preferences for different grape cultivars and their willingness to pay through experimental auctions (Yue et al. 2011) for Minnesota grown grapes, while also determining what fruit attributes are most important to consumers through sensory evaluation. This research will also determine if CSAs and other direct-to-consumer producers would grow their own table grape crops.Marker assisted breeding (MAB)--Grape breeders are turning to MAB to utilize DNA tests to screen both parents and seedlings for desirable traits. MAB is a tool for accelerating the rate of cultivar development by enriching germplasm for the trait of interest and targeting which plants are grown to maturity for evaluation. DNA is extracted from leaf tissue in the greenhouse from a seedling population(s) and extracted using available kits or protocols. The DNA is then subjected testing to determine linkage to the trait of interest to determine which variations were inherited from the parents.MAB is driven by gene mapping experiments that aim to link a trait of interest with a region of a genome. This requires quality phenotyping (trait measurement) across a population in appropriate experimental conditions. These experiments can be in the field, laboratory, or greenhouse. New methodologies that allow for high-throughput phenotyping will accelerate gene mapping and MAB. The use of digital image analysis has proven successful for the mapping the berry color trait and this technology will be considered for other traits like berry size, cluster size, or cluster architecture (Clark et al. 2016).In collaboration with colleagues in the USDA-NIFA funded project VitisGen, over 70 marker-trait associations have been detected. This includes nearly 10 race-specific resistance loci for powdery mildew. These resistance genes are key targets for MAB in breeding for durable resistance because the pyramiding of these genes can only be tested efficiently through DNA tests that demonstrate that favorable copies of the alleles have been combined into a single, durably resistant cultivar.Enology best practices--As part of the breeding program, selected seedlings that show potential during fruit evaluation in the vineyard are processed according to a standardized wine-making procedure, which include control points at important processing steps, with key analyses specified at each step. These procedures allow early assessment of wine quality and comparison of quality across cultivars. Grapes exhibiting acceptable and reproducible wine quality will be propagated to produce larger crops, allowing larger-scale trials of adjusted processing parameters to facilitate and improve production. All wines will be evaluated for standard chemical parameters (pH, titratable acidity, residual sugar, alcohol). Additional evaluations (organic acids, protein and bitartrate stability, tannin assays, volatile acidity, etc.) may be performed as dictated by a particular wine or fermentation method.Nearly all grape cultivars grown in Minnesota are interspecific hybrids developed by plant breeders over the last century. Therefore, unlike varietals from long established winemaking traditions, the best enological practices are still in development. Research will be conducted in order to optimize protocols for cold-hardy cultivars developed (or in development). Modified processing parameter trials will be conducted to develop optimal procedures for individual cultivars. In red wines the effects of skin contact time, fermentation temperature, yeast strains, and tannin/oak on wine quality will be evaluated. In white wines, varying yeast strains, residual sugar, malolactic fermentation, and aging on lees might be performed.Education and extension--Technical information (such as updates on winter injury or disease monitoring) will be provided through electronic media (www.enology.umn.edu). The Minnesota Grape Growers Association (MGGA) and Minnesota Farm Winery Association (MnFWA) newsletter(s) as well as other trade publications will be used to disseminate technical information to grape and wine producers. Direct communication with producers is also important, and continued extension services will create a line of communication among stakeholders and activities conducted at UMN. The project hosts two programs ("Fall Tour" and spring "Research Update") each year which include workshops on techniques, samples of products (grapes or wine), dissemination of research activities, and promotes communication among industry stakeholders. The project also supports the educational efforts of MGGA through speaking at symposium (Cold Climate Conference), organizing events, and regular workshops. Periodic short courses for commercial winemakers and hobbyist have historically been conducted and will continue to be offered.An often underrepresented stakeholder group is hobbyists and home owners who have an interest in growing grapes and making wine. Anecdotally, interest at this level leads to obtaining a winery license or in securing land for a vineyard as individuals become more and more enaged with the opportunities afforded by this expanding industry. In collaboration with the state Master Gardener program, content will be developed in a "train the trainer model" to educate this stakeholder group and have an impact state-wide by utilizing experts who live and share their knowledge about grape growing in their communities.

Progress 07/01/16 to 06/30/21

Outputs
Target Audience:Minnesosta grape growers and wine producers are the target audience for this research project. Research projects extend regionally due to the extensive planting of UMN grape breeding introductions, including most recently 'Itasca', a white wine grape. Additional outputs include collaborations across the University, and with colleagues from across the United States. UMN grape varieties are being tested and introduced in Canada, Europe, Japan, and Israel which also demonstrates the impact of this project on a global scale Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Seven graduate students are directly related with the grape breeding project including Anna Underhill (MS2018) Lu Yin (finished PhD July 2020), Dominique Ebbenga (MS 2018; PhD current), Jack Olson (MS 2020), Davy DeKrey (PhD), Laise Moreira (Phd), and Logan Spicer (MS). One postdoc works directly on the project, studying table grape breeding and evaluation process, Erin Treiber. A second postdoc, Kate Freund, screened several mappign populations of grape (along with other species) for natural resistance to Japanese beetle screening. This is a collaboration with Drs. Adrian Hegeman and Stan Hokanson. Additionally, graduate student Stephen Brockman has been investigating floral aromas in grapevine. Several undergraduate students have worked on this project over the course of the year but were cut short due to the work-from-home restricts of the pandemic. The students were engaged in multiple aspects of the project including genetic analysis, embryo rescue, data entry, and plant are. In February 2020, two members of the team PI Clark and graduate student Laise Moreira presented their research projects at the International Table Grape Symposium held in Santiago, Chile. This was a profressional development activity and excellent intorduction to table grape production on a global scale. How have the results been disseminated to communities of interest?Results have been disseminated through various channels including: peer reviewed journals, industry and academic meetings, grower field days, and grower workshops. Additionally, progress reports are given annually at the Cold ClimateConference to assembled industry stakeholders. Research is also translated into Extension products including programs, workshops, publications, and posted on our website enology.umn.edu What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1. Advance hybrid grape cultivar development for (a) cold-hardy wine grape production, (b) table grape markets, (c) food, beverage, and food product industry, and (d) ornamental landscape installations through traditional and high-throughput phenotyping methods. 2. Implement routine marker assisted breeding (MAB) in grape cultivar development through (a) identifying marker-trait associations, (b) developing DNA tests for associated traits, (c) implementing DNA tests for screening potential parents and culling offspring. 3. Develop protocols and best enology practices for commercial wine production using cold-hardy grape cultivars. 4. Advance the understan

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Moreira, L. and M. Clark (2021) Embryo rescue of cold-hardy table grapes. HortScience 56(9): 1059-1065
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Yin, Lu, Avinash Karn, Lance Cadle-Davidson, Cheng Zou, Anna Underhill, Paul Atkins, Erin Treiber, Daniel Voytas, and Matthew Clark. "Fine mapping of leaf trichome density revealed a 747-kb region on chromosome 1 in cold-hardy hybrid wine grape populations." Frontiers in plant science 12 (2021): 150.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Yin, Lu, Eric C. Burkness, William D. Hutchison, and Matthew D. Clark. "Effects of Foliar Phylloxera (Hemiptera: Phylloxeridae) Infestations on Wine Grape Photosynthesis, Yield, and Fruit Quality." Journal of Entomological Science 56, no. 4 (2021): 504-518.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Olson, Jack, and Matthew Clark. "Characterization of Anatomical and Physiological Effects of Variegation Mutation on Grapevine." HortScience 56, no. 10 (2021): 1251-1257.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Minnesosta grape growers and wine producers are the target audience for this research project. Research projects extend regionally due to the extensive planting of UMN grape breeding introductions, including most recently 'Itasca', a white wine grape. Additional outputs include collaborations across the University, and with colleagues from across the United States. UMN grape varieties are being tested and introduced in Canada, Europe, Japan, and Israel which also demonstrates the impact of this project on a global scale. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Six graduate students are directly related with the grape breeding project including Lu Yin (finished PhD July 2020), Dominique Ebbenga, Jack Olson (completed MS Sept 2020), Davy DeKrey, Laise Moreira, and Logan Spicer. One postdoc works directly on the project, studying table grape breeding and evaluation process, Erin Treiber. A second postdoc, Kate Freund, screened several mappign populations of grape (along with other species) for natural resistance to Japanese beetle screening. This is a collaboration with Drs. Adrian Hegeman and Stan Hokanson. Additionally, graduate student Stephen Brockman has been investigating floral aromas in grapevine. Several undergraduate students have worked on this project over the course of the year but were cut short due to the work-from-home restricts of the pandemic. The students were engaged in multiple aspects of the project including genetic analysis, embryo rescue, data entry, and plant are. In February 2020, two members of the team PI Clark and graduate student Laise Moreira presented their research projects at the International Table Grape Symposium held in Santiago, Chile. This was a profressional development activity and excellent intorduction to table grape production on a global scale. How have the results been disseminated to communities of interest?Results have been disseminated through various channels including: peer reviewed journals, industry and academic meetings, grower field days, and grower workshops. Additionally, progress reports are given annually at the Cold Climate Conference to assembled industry stakeholders. Research is also translated into Extension products including programs, workshops, publications, and posted on our website enology.umn.edu. What do you plan to do during the next reporting period to accomplish the goals?We will continue progress on each of these goals. We have been quite successful with implementation. We do plant to reduce the number of seedlings for evaluation due to limitations in the nursery and to target specific traits. Cultivar development and testing continues to proceed and plans are in place for the release of new varieties with disclosures made within appropriate University channels.

Impacts
What was accomplished under these goals? 1. In 2020 we planted nearly 8,000 seedlings into our seasonal nursery for initial evaluation (pest and disease resistance, and vigor). We also submitted nearly 2,000 of these seedling for marker assisted breeding (goal 2). From these ~8,000 plants we have selected approximately 1,200 for advancement with traditional phenotypic selection. Several populations have known disease resistance, for example Run1. These populations segregated phenotypically in the nursery for powdery milew resistance and thus were easily culled. The nursery includes both wine and table grape seedlings. In addition to the nursery grown seedlings, embryo rescue was employed for the development of seedless table grapes. For this growing season, several hundred embryo rescuederived seedlings were planted directly in to the vineyard for establishment, thus skipping a year in the nursery screening. This is due to the fact that plants were much larger due to germination initiated in November and the seedlings were greenhouse grown for several months. In 2020, we implemented more digital data collection for field observations of seedlings and advanced selections. In particulary we used the applciation "Fieldbook" and had support from the Breeding Insight group (Cornell/USDA). This has allowed us to have better access to multi-year data for making culling decisions in the field. This is the fourth year for which we have used digital image analysis for aiding in genetic studies and record keeping. Images of grape clusters have been captured for many of the grape mapping populations for genetic studies as well as table grape and advanced wine grape selections. Images can be processed to extract data such as color, cluster compactness, and bery size. In 2020, we got the first reports of table grape testing from off-site collabortator/testers. We also continue to identify testers for all grape selections using the NE1720 network or through growers who are interested in testing our products. The project has also made some crosses specifically for ornamental landscape uses. These include those with red/purple foliage all season, or red foliage in autumn. The other interesting trait called "parsley leaf" has been introgressed into our materials from the variety Chasselas Ciotat and became part of a genetics study with collaborators at North Dakota State University. We shared seeds from targeted crosses for use in understanding the genetics of this trait and the project is ongoing. A third trait that is of itnerest is leaf variegation. A graduate stude on the project, identified a major genomic region associated with this trait using QTL mapping approaches. 2. Marker assisted breeding has become a critical part of the breeding cycle in our program. Graduate students have been focused on identifyign marker trait associations and have been extraordinarily successful for the traits: trichome density/type, leaf variegation, and insect resistance (foliar phylloxera). We have used a fine-mapping approach by establishing a large, 1000 individual, popualtion for the expressed purpose of narrowing the QTL region for important traits. This population has also been used for better understanding of powdery mildew resistance in collaboration with colleagues at USDA-ARS in Geneva, NY and Cornell University. We have implemented rhAmpSeq markers in 2020, but due to covid the marker data has not been completely analyzed at the time of this report. The whole-genome marker platform allows the researcher to target loci of interest for routine MAB, but is affordable for whole-genome mapping of populations. A new population introduced into the program this year will be used to study Ren10 powdery mildew resistance which was first identifed several years ago at UMN. 3. Enology specialist Drew Horton continues to develop protocols and test enology practices related to UMN varities and other important varities grown in the region. As part of a a Specialty Crop Block Grant, our project in collaboration with NDSU have studied the development of grapes (Juice) and wine from SE Minnesota along a transect which into NW Minnesota and into North Dakota. This "terroir" project aims to document ripening along this transect and included additional vareities than previously proposed. We also extended this research to investigate the same basic question about ripening in 'Itasca' grape at the Horticutlural Research Center in 2 years. Current recommendations for most wine grape varities is to also grapes to mature on the fine for an extended period to reduce the acidity in the must. However, with low-acid grapes, this practice may actually result in changes to the wine chemistry (elevated pH) or produce off flavors and aromas. This is known in other varieties like 'Brianna'. This type of research is critical for the sucess of a new variety when growers are only getting the first commericial harvests in recent (and upcoming years) 4. Due to Covid-19, efforts in this area were greatly reduced after March 2020. However, our project along with collboration of Annie Klodd (Extension Educator) supported the Minnesota Grape Growers annual Cold Climate Conference in Feburary. Were were critical to the success of the programming provided and organization of the conference, in genera. Drew Horton led workshops on filtration (and other topics) but was able to partner with a nearby winery outside Rochester, MN to cover topics that are typically difficult to convey in the classroom setting. Through spring and summer, we partnered with the University of Wisconsin-Madison Extension specialists in horticultural crops and entomology to produe a well attended webinar series. Additionally we provide 'consulting-like' services over the phone, emails, and in grower visits as needed.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2020 Citation: Ebbenga, D. N.*, Burkness, E. C., Clark, M. D., & Hutchison, W. D. Risk of increased volatile acidity and Spotted wing drosophila injury in Minnesota wine grape varieties. American Journal for Enology and Viticulture.
• Type: Journal Articles Status: Accepted Year Published: 2020 Citation: Underhill, A. N.*, Clark, M. D., & Hirsch, C. D. Image-based phenotyping identifies QTL for cluster compactness in grape. J ASHS.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Zou, C., Karn, A., Reisch, B., Nguyen, A., Sun, Y., Bao, Y., Cambell, M. S., Church, D., Williams, S., Xu, X., Ledbetter, C. A., Patel, S., Fennell, A., Glaubitz, J., Clark, M., Ware, D., Londo, J., Sun, Q., & Cadle, L. (2020). Haplotyping the Vitis collinear core genome with rhAmpSeq improves marker transferability in a diverse genus. Nature Biotechnology.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Underhill, A. N.*, Clark, M. D., & Hirsch, C. D. (2020). Evaluating and mapping grape color using image-based phenotyping. Plant Phenomics: 8086309. https://doi.org/10.34133/2020/8086309
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Maher, M. F.*, Nasti, R. A.*, Vollbrecht, M.*, Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene editing through de novo induction of meristems. Nature Biotech. doi: https://doi.org/10.1038/s41587-019-0337-2
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Yin, L.*, Clark, M., Burkness, E., & Hutchison, W. (2019). Grape Phylloxera, Daktulosphaira vitifoliae Fitch (Hemiptera: Phylloxeridae), on Cold-hardy Hybrid Wine Grapes (Vitis spp.): A Review of Pest Biology, Damage, and Management Practices. J. Integrated Pest Management. doi: https://doi.org/10.1093/jipm/pmz011
• Type: Journal Articles Status: Submitted Year Published: 2020 Citation: Yin. L., Burkness, E., Hutchison, W.D., & Clark, M.D. Effect of foliar phylloxera infestation on grapevine photosynthesis, yield, and fruit quality. [Submitted to J. of Entomological Science]

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Minnesota grape growers and wine producers are the primary target audience. Research outputs extend regionally to other states that grow and produce wines of the same cold-hardy, grape varieties. Additional outputs include international collaborations and the expansion of UMN grape varieities into Canada, Europe, Japan, and Israel. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Six graduate students have focused on grapevine evaluation in this reporting period including Lu Yin, Laise Moreira, Anna Underhill, Dominique Ebbenga, Jack Olson, and Davy DeKray. We also have a collaboration with Dr. Adrian Hegeman and one of his PhD students Stephen Brockman has used grape as a model for some of his research. Undergraduates also support the project and conduct research in various areas. These include Alex Cummings, Abigail Diering, Killian Harnish, and Anna Andresen. Students have focused on different projects including: mass spectrometry of aromatic compounds; sensory evaluation of wine and table grapes; evaluation of table grapes for disease, post harvest, and quality traits; digital image analysis of grape clusters; development of digital collection for field and sensory data; and genetic determinants of variegation in grape seedlings. How have the results been disseminated to communities of interest?Results have been disseminated through various channels including: peer reviewed journals, industry and academic meetings, grower field days, and grower workshops. Additionally, progress reports are given annually at the Cold Climate Conference to assembled industry stakeholders. Research is also translated into Extension products including programs, workshops, publications, and posted on our website enology.umn.edu. What do you plan to do during the next reporting period to accomplish the goals?We will continue to progress on each of the goals. The graduate students and post-doc will continue to focus on their specific research areas. Cultivar development continues to proceed. We have plants for marker assisted selection of seedlings in spring 2020.

Impacts
What was accomplished under these goals? 1. Cultivar development is a key component of this project. In this reporting season we have evaluated nearly 3,500 seedlings with DNA tests to screen for powdery mildew resistance (Ren3, Ren9, Ren10, Run1). We have been able to pyramid some resistance genes using our DNA testing procedures. Additionally, we continue to make crosses ~100 per year to maintain our pipeline of seedlings for evaluation. Approximately 10,000 seeds were produced in Spring 2018. These are tested with DNA markers as appropriate, culled, and then evaluated in the nursery. About 1500 of these will be planted into the vineyard in 2020. Several advanced lines have been identified for further testing and have been distributed to Unviersity collaboratores (NE1720 project) as well as grower-testers, and nurseries. We have recieved additional funding for the evaluation of table grapes. 1b)Additional effort has been placed into embryo rescue techniques for the advancedment of seedless variety development. We are currently evaluating ~30 selections for harvest fruit quality and post harvest quality evaluations. This information will be useful in setting breeding selection criteria and in identifying potential new cultivars. 1c)In this reporting period an undergraduate student completed a directed studies project to investigate a single seleciton for use in food product industry. This report has been used for engaging with food scientists at the University of Minnesota, and for developin protocols for evalulation of this selection and other suitable selections. 2. Marker assisted breeding using the AmpSeq and rhAmpSeq pipeline is ongoing in collaboration with the USDA-SCRI funded VitisGen2 project. All testing has been conducted in this collaboration. Graduate and undergradate students have been engaged in genemapping studies to identify the genetic regions (loci) that are underlying key traits including: berry color, cluster shape, volatile compounds, leaf variegation, leaf trichomes, and insect resistance.We have collaborated with colleagues from USDA-ARS and Cornell University to breed durable resistant selections (those with stacked powdery mildew resistance genes). The use of DNA marker tests are required for this type of work. The plant material that is selected will be made available publicly to improve grape breeding and the time to cultivar release. 3. In this reporting period we conducted wine making practices for 'Marquette' and 'Itasca' grapes. These were evaluated using sensory studies. A full report can be found here:https://enology.dl.umn.edu/wine/sensory-analysis-itasca-wines-preliminary-report. Nearly 100 selections were evaluated for juice chemistry traits as part of the breeding program. Approximately 75 wines were produced in fall 2018 and were evaluated by the project team in spring 2019. This information was used for making breeding and selection decisions. Currently we are doing a project with North Dakota State University (Hatterman-Valenti) to evaluate the role of terroir on Marquette, La Crescent, and Brianna grapes grown across the region. Fruit have been harvested and wine produced during this reporting cycle. The wines will be evaluated by industry and sensory panels. 4. In collaboration with Annie Klodd, Extension educator, and enology specialist Drew Horton, we have been producing workshops, webinars, and wine maker round tables to engage the grower/winemaker community. We have all presented at the annual Cold Climate Conference on research updates. We facilitated a round table discussion with growers at the event. Additionally we have hosted and sponsored on-the-farm workshops on viticultural techniques with three different growers as part of a SARE partnership grant.

Publications

• Type: Book Chapters Status: Published Year Published: 2019 Citation: Clark, M.D. 2019. Development of Cold Climate Grapes in the Upper Midwestern U.S.: The Pioneering Work of Elmer Swenson. In Plant Breeding Reviews Vol 43. Ed. I. Goldman. p. 31-59
• Type: Journal Articles Status: Submitted Year Published: 2019 Citation: Maher, M.F., Nasti, R.A., Vollbrecht, M., Starker, C.G., Clark, M.D., & Voytas, D.F. Plant gene editing through de novo induction of meristems. Submitted Nature Biotech
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moreira, L.S., Underhill, A., and M. Clark. Postharvest evaluation of cold-hardy table grape breedling lines. XII International Conference on Grapevine Breeding and Genetics. .doi:10.17660/ActaHortic.2019.1248.15
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Fennell, A., Alahakoon, D., Luby, J. and Clark, M. (2019). Mapping the genetic architecture of grapevine bud growth-cycling quantitative traits. Acta Hortic. 1248, 337-344 DOI: 10.17660/ActaHortic.2019.1248.49 https://doi.org/10.17660/ActaHortic.2019.1248.49
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Rice, S. Tursumbayeva, M., Clark, M., Greenlee, D., Dharmadhikari, M., Fennell, A., and J.A. Koziel. 2019. Effects of harvest time on aroma of white wines made from cold-hardy Brianna and Frontenac gris grapes using headspace solid-phase microextraction and gas chromatography-mass spectrometry-olfactometry. Foods. 8(1): 29. https://doi.org/10.3390/foods8010029
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Yin, L., Clark, M.D., Burkness, E.C., & Hutchison, W.D. (2019) Grape phylloxera (Hemiptera: Phylloxeridae), on cold-hardy hybrid wine grapes (Vitis spp.): A review of pest biology, damage, and management practices

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Minneosta grape growers and wine producers are the primary target audience. Research outputs extend regionally to other states that grow and produce wines of the same cold-hardy, grape varieties. Changes/Problems:No major changes to report. What opportunities for training and professional development has the project provided?Three graduate students have worked on the grape breeding project including Lu Yin, Laise Moreira, and Anna Underhill. A graduate student in Plant Entomology is also involved in the project (Dominique Ebbenga). Several undergraduate students worked on the project including: David Tork, Abigail Diering, Jack Olson, Grace Watson, Stein Iverson, and Anna Andresen. Each student has focused on a research area including: mass spectrometry, automation of data collection, and evaluation of variegation in grape seedlings. Enology Specialist Drew Horton participated in 2 wine competitions. At "Indy" he was an observor and judge in training. He was an invited judge for the "Jefferson Cup". His participation brings value to the project and to the International Cold Climate Wine Competition co-sponsored by this project. How have the results been disseminated to communities of interest?Results have been disseminated through peer reviewed manuscripts (see above). Also through field days, presentations at the Cold Climate Conference (hosted by the Minnesota Grape Growers Associations), through the enology.umn.edu website, one-to-one communication, stakeholder board meetings, webinars, and workshops. What do you plan to do during the next reporting period to accomplish the goals?We will continue to work on each of these goals. Three graduate students are engaged in the program to support the advancement of marker-assisted selection and to drive the breeding program towards improved cultivars for fruit quality traits (including seedlessness) and disease and pest resistance. We have a new collaborative project with North Dakota State University for evaluating the effects of terroir in the variety Brianna, on both vine growth and architecture, and wine production

Impacts
What was accomplished under these goals? 1. Plants of UMN varieties and other cold-hardy hybrids continues in Minnesota and in the region. The recent release of 'Itasca' in 2017 has mean that more acerage has been installed for this variety. Additionally, some growers are replacing older cultivars with new ones. The breeding program planted nearly 7,000 seedling vines for evaluation in the nursery and ~1,200 vines into the vineyard for long term evaluation. Several advanced lines were considered for testing in multi-state evaluation (NE1720). 1b. We are continuing to trial table grapes and invest in table grape breeding. We have utilized the embryo rescue technique for the second year. This process allows us to make seedless x seedless crosses and results in hundreds of new seedlings for evaluation. We have partnered with grower and research groups to test several of the advanced table grape lines. Graduate Student Anna Underhill has focused on developing high-throughput phenotyping methods. Primarily the use of digital image analysis of berry clusters. Anna has been developing platforms and protocols to characterize bunch compactness. Additionally, our group has focused on fruit color and fruit chemistry using various methods including: digital image analysis, mass spectrometry, 96-well plate reader (color and enzymatic analysis of fruit quality traits like malic acid), and the use of other sensors. 2. In 2018, we utilized AmpSeq technology to screen our seedlings: Extracted DNA from ~3897 seedlings: Screened about 1726 for culling as not all seedlings submitted ended up segregating for the traits of interest. Also we had a large population for fine mapping. Targeted three loci: SDI (seedless); Ren3 (identified individuals potentially fixed at this locus such as 'Itasca'); and Muscat Eliminated ~ 963: kept~44% Have Rampseq on ~1000 individuals for fine mapping population. Identified recombinants at the LG14 Phylloxera QTL. Cloned ~200 lines for a replicated greenhouse experiment which is being conducted by graduate student Lu Yin. 3.In 2017, nearly 100 research wines were produced and evaluated by the research staff for fruit quality. We also conducted a small yeast trial. We produced 'itasca' wines with four different yeast, and a second treatment of malo-lactic fermentation. in March 2018, at the Minnesota Grape Growers Association Cold Climate Conference ~70 attendees participated in a sensory evaluation of the 8 wines. We used this opportunity to teach growers about the wine selection, the malo-lactic fermnation process. Our preliminary evaluation of the wines indicated no consumer preference among the treatments. Additional experiments conducted in early 2018 include whole cluster Marquette wine production and evaluation of skin contact treatments for flavor development in Itasca wines. 4. Annie Klodd, Extension Educator and Dr. Clark have strengthened relationships with the grower and wine producer community. We hosted Dr. Richard Smart for a workshop. The topic of the workshop focused on trunk disorders in cold climate grapes. We have used the opportunity to engage with many growers. We have also hosted and presented webinars, field days, and research updates to grower/producer audiences. Drew Horton, enology specialist hosted wine-maker round tables monthly for wine producers.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Clark, M., Teh, S. L., Burkness, E., Moreira, L., Luby, J., Watson, Yin, L., G., Hutchison, W., (2018). QTL Identified For Foliar Phylloxera Resistance in A Hybrid Grape Population. Australian Journal of Grape and Wine Research 24(3): 292-300.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Yin, L. (2018) Characterization of leaf trichome for its potential role in resistance to foliar phylloxera in a cold-hardy hybrid grape population (e-poster & Student Ignite Competition). American Society of Horticultural Science.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Underhill, A., C. Hirsch, and M. Clark. 2018. Image-based phenotyping and genetic control of cluster density traits in interspecies grapevine hybrids. Poster presented at: Plant & Animal Genome XXVI, San Diego, CA. 13-17 Jan. Poster P0644.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Underhill, A., A. Diering, D. Tork, D. Freund, A. Hegeman, S. L. Teh, and M. Clark. 2018. Multiple phenotyping methods find QTL for berry color in an interspecific hybrid grape (Vitis spp.) population. Paper presented at: Taming High pH in the East. American Society for Enology and Viticulture-Eastern Section 43rd Annual Conference, King of Prussia, PA. 9-11 July.
• Type: Other Status: Published Year Published: 2018 Citation: Clark, M., B. Tuck, and A. Klodd 2018. 2017 Minnesota Grape Production Statistics. ESTIMATES FOR THE YIELD, PRODUCTION, AND PRICING DATA OF THE MINNESOTA GRAPE INDUSTRY. University of Minnesota Extension. https://enology.dl.umn.edu/sites/g/files/pua1676/f/media/2017_minnesota_crush_report.pdf

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Minnesota grape growers and wine producers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Three graduate students have worked on the grape breeding project including Lu Yin, Laise Moreira, and Anna Underhill. A graduate student in Plant Entomology is also involved in the project (Dominique Ebbenga). Two undergraduate students worked on the project including David Tork (color analysis) and Hannah Hanlon (wine and juice chemistry analysis). One summer worker, Nicole Bacheller, worked in both the vineyard and winery. A summer intern through the LSSURP program, Daphne Cobbs, worked on identifying SRAP markers linked to the variegation trait in seedling grapevines. How have the results been disseminated to communities of interest?Results have been disseminated through peer reviewed manuscripts (see above). Also through field days, presentations at the Cold Climate Conference (hosted by the Minnesota Grape Growers Associations), through the enology.umn.edu website, one-to-one communication, stakeholder board meetings, and workshops. What do you plan to do during the next reporting period to accomplish the goals?We will continue to work on each of these goals. Three graduate students are engaged in the program to support the advancement of marker-assisted selection and to drive the breeding program towards improved cultivars for fruit quality traits (including seedlessness) and disease and pest resistance.

Impacts
What was accomplished under these goals? 1. This year we worked towards all subgoals in this objective. (a) we released the new white wine cultivar 'Itasca' for sale in 2017. Reports indicate that ~40,000 vines were sold and distributed acrossed the United States. We are working to obtain variety protection in Canada so that we can also import virus free materials in 2017/2018 to licensed producers. Additionally, we have continued the decades-old grape breeding program. In 2017 we started nearly 7,000 seedling vines. These were grown in the greenhouse and a subset evaluated with DNA markers for disease resistance. Following several culling steps, ~2000 of these vines will be moved forward for further testing. One graduate student is developing digital image analysis techniques to measure grape clusters for important traits like color, compactness, berry size, etc.(b) We have invested heavily in embryo rescue techniques and personell to advance table grape research. One technican, now a PhD student, is evaluating the best methods for this specialized technique to advance the germplasm. Several seedless table grape lines have gone out on testing agreements for further grower evaluation. (c) We have done small scale, pilot experiments on vinegar production using current UMN cold hardy varieities as well as advanced selections. These will be evaluated in 2017 for suitability for advanced study and use in exploring other grape products. (d) Ornamental grapevines have a role in many landscapes. We have produced hybrid crosses for evaluation that are intended to be disease resistant, have color fall foliage, and unique leaf shapes. 2. (a) This year we have worked on several traits to associated markers with breeding targets. We have mapped a major QTL for phylloxera resistance to chromosome 14 using field and greenhouse experiments. Phylloxera is a major insect pest of grapes world wide. We are working on developing tightly linked markers that can be used to screen seedlings for this trait. We also had a summer intern who worked on a variegation trait we observe in 'Frontenac' offspring which appears to be a recessive trait. The intern has worked to identify a new marker platform for use in bulk segregant analysis. (b) This objective is ongoing. We are currently working on improving current markers and developing new ones for traits of interest. This includes SNP based and sequencing techniques like AmpSeq. (c) In 2017 we screened nearly 5000 plants with DNA markers for powdery mildew resistance. 3. The enology specialist evaluated 2 wine making techniques at harvest in 2016including whole cluster Marquette and malolactic fermentation in the white wine 'La Crescent'. These wines were evaluated in internal sensory panels and then presented at the annual UMN Grape Breeding and Enology Project research update in March 2017 to industry stakeholders. In 2017 we are conducting a yeast and malo-lactic fermentation on the 'Itasca' grape and will present these wines at the Cold Climate Conference in March 2017 for sensory evaluation. 4. We have conducted a number of outreach and extension activites in this reporting document. The project hosted 2 "field day" events that draw in >50 attendees. In September we have a traditional field day and present information about the varieties and have a guest speaker. This year Dominque Ebbenga presented her work on spotted wing dropsophila. In March we hosted our research update, in which graduate students, staff, enology specialist, and I present an update on the current research projects, grant proposals, as well as preliminary and final results. We discuss wine making practices as well as vineyard practices, principally cold hardiness evaluation of lines following winter. In addition to these field days Drew Horton has conducted wine-maker round tables that are open to wine makers and winery operators. This has been a very successful extension activity. Participants trial the wines "blind" and provide constructive feedback about what they are tasting as well as provide the details about the winemaking practices. After the first event, one attendee indicated "I know I need to change what I am doing" and now attends each session and is making efforts to improve wine quality. In 2017 we produced the first crush report for Minnesota that captured harvests from 2013-2016 to get a baseline for the amount of production in the state as well as the prices paid for the grapes. The survey also documented losses, and their sources so that we can continue to target breeding efforts as well as extension programming to increase the sustainability of grape production.

Publications

• Type: Books Status: Published Year Published: 2017 Citation: Minnesota Grape Growers Association. 2016. Growing Grapes in Minnesota (10th Ed.) revised by P. Domoto, C. Anderson, M. Clark, and I. Geary.
• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Teh, S.L., Fresnedo-Ramirez, J., Clark, M.D., Sun, Q., Cadle-Davidson, L., and J.J. Luby. 2017 Genetic dissection of powdery mildew resistance in intersp
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Fresnedo-Ram�rez, J., S. Qi, C.-F. Hwang, C.A. Ledbetter, D.W. Ramming, A.Y. Fennell, M.A. Walker, J.J. Luby, M.D. Clark, J.P. Londo, L. Cadle-Davidson, G.-Y. Zhong, and B.I. Reisch. 2016. Toward the elucidation of cytoplasmic diversity in North American grape breeding programs. Mol. Breeding 36: 116. doi:10.1007/s11032-016-0538-z
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Clark, M., Hemstad, P, and J. Luby. 2017. Itasca Grapevine, a New Cold-hardy Hybrid for White Wine Production. HortScience 52 (4): 649-651.
• Type: Other Status: Published Year Published: 2017 Citation: Clark, M and B. Tuck. 2017. Minnesota Grape Production Statistics: 2013-2016. https://www.extension.umn.edu/community/economic-impact-analysis/reports/docs/2017-mn-grape-production-stats.pdf

Progress 07/01/16 to 09/30/16

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1) Developed methods for screening foliar phylloxera infestation in the greenhouse. Implementing digital image analysis for berry color 2) Developing molecular markers for seedlessness and for disease resistance. Identified major QTL for phylloxera resistance 3) Hired enology specialist. Implementing wine making trials in 2016 crush season. 4) Continue to provide updated information and respond to wine makers and grape growers through website (enology.umn.edu) and through google group for Minnesota grape growers association and Minnesota Farm Winery Association Members.

Publications