Progress 01/15/21 to 01/14/24
Outputs
Target Audience:Target Audiences:This project targeted national and international students at the undergraduate and graduate levels that have an interest in applied breeding and genetics, cereal grains seed industry representatives that actively develop improved crop products for farmers, crop producers, and academic faculty that have current or had previous research programs that involve plant breeding, crop genetics, and/or general plant sciences. There was a focus early in the project to interact with collaborating scientists and communicate project deliverables. A postdoctoral research fellow (Dr. Carolina Ballen-Taborda; female minority) was hired in year 1 (June 2021) that had a background in plant genetics, genetic mapping, trait introgression, and bioinformatics. Masters student Mary Frances-Behnke was onboarded in August 2021 and graduated in August 2023. In addition to training early career plant breeders and geneticists, wheat industry stakeholders were the primary audience of the project with the development and testing of advanced wheat cultivars. Traits such as yield and test weight that directly impact wheat producers were of highest priority, but Fusarium head blight resistance was another focal trait to increase soft wheat quality in the region that directly benefits millers and livestock producers by reducing mycotoxin (e.g., deoxynivalenol) levels in the wheat supply chain. This breeding objective indirectly supported the general public, particularly consumers of wheat-based products and meat-based proteins produced using wheat, by helping to provide safe and healthy wheat-dependent products in regional and national markets. Project personnel delivered applied research information to the scientific community that was specifically aimed to support new or emerging breeding programs. Research results were routinely and directly shared with institutional breeders within the Southeastern University Small Grains Breeding Cooperative (SunGrains). Efforts:A plant breeding workshop was hosted by Clemson University (PI Boyles was a co-organizer and co-host) virtually in June 2021 that was free to register and attend. This effort facilitated more than 200 individuals to attend and learn about conventional and molecular breeding practices across numerous important field crops. This four-day event contained presentations from nationally and internationally renowned scientists in breeding, genetics, biofortification, and human nutrition. A spring nursery tour was hosted by PI Boyles at the Pee Dee Research and Education Center in April 2021. Attendees were Dr. Paul Murphy (Professor, NC State), Dr. Robert Etheridge (President, Mixon Seed), Jimmy Clements (President, AgSouth Genetics), and Blake Shephard (Mixon Seed). To accommodate safe practices during the COVID-19 pandemic, individual meetings with seed industry representatives were facilitated by PI Boyles during spring 2021 that enabled representatives to visit the wheat field site, observe advanced breeding lines, and discuss line performance with PI Boyles. Companies that had representatives visit included Gerard Seed Co, JoMar, KWS Cereals, Limagrain Cereal Seeds, and NorthStar. PI Boyles attended a field day hosted by Mr. Carl Coleman (wheat producer and President of Little Mill Seed) on behalf of GrowPro Genetics (formerly Syngenta Apri Pro) in Dillon, SC in May 2021 to meet with various extension agents, wheat producers, and industry representatives.PI Boyles also gave a presentation on advanced breeding lines and likely future cultivar releases at a GrowPro Genetics wheat field day event in Dillon, SC in May 2022, as well as at the NC Small Grain Growers Association annual meeting in Durham, NC in January 2023. PI Boyles visited six different small grains farm operations in the reporting period to gain feedback on critical production challenges that could be addressed through plant breeding and genetics. In addition, feedback was received from crop consultants and private companies that license small grains releases from the Clemson program to understand how well varieties performed and if particular improvements can be made in future cultivars. In February 2022, PI Boyles participated in the USDA NIFA Wheat Resiliency Conference led by the University of Kentucky to share his vision with colleagues on how to effectively prepare the wheat crop for impending climate change. PI Boyles attended the 2022 National Fusarium Head Blight Forum in Tampa, FL to provide updates on research progress to collaborators within the Variety Development and Host Resistance (VDHR) Southern Winter Wheat (SWW) coordinated project, which is supported by the US Wheat & Barley Scab Initiative. In December 2023, PI Boyles gave an invited presentation at this same National FHB Forum, which was held in Cincinnati, OH, to highlight historical achievements in wheat improvement as well as inform the audience about recent progress made by the Clemson (and broader SunGrains cooperative) wheat breeding program. At these meetings, PI Boyles provided regional updates on wheat industry personnel and production changes to the National Wheat Improvement Committee (NWIC). Postdoc Carolina Ballen-Taborda presented project research findings at the 2022 ASA, CSSA, SSSA International Annual Meeting in Baltimore, MD, and Dr. Ballen-Taborda also placed third in a university-held poster competition held in 2022 by the Clemson University Postdoctoral Association. Lead breeders from the seven SunGrains breeding programs (Arkansas, Clemson, Florida, Georgia, LSU, NC State, and Texas A&M) met biannually each year with the spring field tours rotating across 2-3 states (2021: NC and SC; 2022: LA and TX; 2023: FL, GA, and SC) and a late summer meeting for planning the upcoming season research activities. The late summer meeting provided time to discuss project genotyping plans such as number of lines per program and deadlines for sampling and sequencing. A wheat field day was organized and hosted by PI Boyles in April 2023 at the Pee Dee Research and Education Center in Florence, SC. Attendees of the event included wheat growers from NC and SC, seed company representatives, animal production company representatives, chemical company representatives, regional small grains breeders, and university extension agents. Speakers at the event included Robert Etheridge (President of Mixon Seed), Dr. Christina Cowger (USDA-ARS Raleigh), Alex Coleman (Clemson small grains extension specialist), Dr. Mike Marshall (Clemson weed scientist), and PI Boyles. The project team published 15 journal articles, a book chapter, and gave 12 presentations to disseminate research to the scientific community. Another three manuscripts are in various stages of the publication process to further increase outreach, and a second wheat field day is scheduled for April 2024 to update wheat producers and industry stakeholders on final project products, particularly new wheat varieties released or nearing release. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Postdoctoral Research Associate: Clemson University postdoc Dr. Carolina Ballén-Taborda, with a PhD in Plant Breeding, Genetics, and Genomics from the University of Georgia, completed the implementation and optimization of genomic prediction models for the South Carolina region, by incorporating large historical genomic and phenotype data generated and maintained by SunGrains. Dr. Ballén-Taborda was educated in year 1 on genomic prediction strategies that are currently implemented in Dr. Boyles' Cereal Grains Breeding & Genetics (CGBG) program. She successfully learned about the datasets and bioinformatics pipeline that are used to facilitate generating genomic estimated breeding values (GEBVs) for individual wheat lines at the preliminary testing stage. Her research on implementing genomic prediction was published in Frontiers in Genetics journal (Ballén-Taborda et al 2022) and presented at the ASA-CSSA-SSSA Crop and Soil International Annual Meeting in November 2022. Furthermore, Dr. Ballén-Tabordaworked on performing simulation of progeny performance using the PopVar tool. This has included two different research projects: 1) 2-year PopVar predictions and field validation, and 2) retrospective PopVar analysis to study whether this tool could allow breeders to develop new cultivars by focusing on promising parental combinations. This research was presented at an on-campus venue in 2021 (the 1st Annual Postdoctoral Symposium) and will be presented in January 2023 at the American Seed Trade Association (ASTA) - 62nd Vegetable & Flower Seed Conference. In addition, Dr. Ballén-Taborda was able to acquire hands-on learning by supporting seed processing, preparing wheat for planting, planting, selection, and harvest in every project field season. Dr. Ballén-Tabordaimproved her data analysis and programming skills in R and was trained and advised directly byPDBoyles and Dr. Jeanette Lyerly from NS State. Finally, sheparticipated in SunGrains meetings to interact with wheat breeders and attended interactive workshops and trainings in R and Linux with the Clemson Computing and Information Technology group (CCIT). Graduate students:Graduate students AJ Ackerman (PhD earned in August 2022) and Mary-Frances Behnke (MS degree receivedin August2023) were involved with wheat cultivar development stages such as greenhouse crossing, early generation bulk field selections, headrow evaluation, data collection in yield plots, combine and headrow harvest, and post-harvest grain processing. Both have contributed to evaluating wheat for Fusarium head blight resistance, which Dr. Ackerman recently publishedon this effort (Ackerman et al. 2022 Agronomy). Dr. Ackerman successfully landed a postdoctoral research position at the University of Illinois Urbana-Champaign with the wheat breeding and genetics program and since took a position with Breeding Insight at Cornell University. Ms. Behnke attended the 2023 Plant and Animal Genome Conference (13-18 January) to presenther research findings. Ms. Behnke is now a PhD student in the Wallace Lab (wallacelab.uga.edu)at the University of Georgia-Athens. Project personnel group trainings:Practical Breeding Concepts weekly meetings trained postdocs and graduate students in the CGBG program on relevant topics in applied plant breeding that are difficult to learn through traditional coursework. Practical Breeding Concepts meetings led by PD Boyles provided environments for discussing existing program initiatives as well as brainstorming about new ideas and methodologies to improve upon the cultivar development process and accelerate genetic gain. During early months of each calendar year (January-March), scientists in SunGrains breeding programs collectively met remotely each week as part of a journal club ("Plant Breeding Gumbo") session where a SunGrains member would present a research paper focused on an emerging topic in wheat breeding and genetics. Personnel from SunGrains breeding programs (Arkansas, Clemson, Florida, Georgia, LSU, NC State, and Texas A&M) met biannually each year with the spring field tours rotating across 2-3 states (2021: NC and SC; 2022: LA and TX; 2023: FL, GA, and SC) and a late summer meeting for planning the upcoming season research activities. The late summer meeting provided time to discuss project genotyping plans such as number of lines per program and deadlines for sampling and sequencing. Project personnel professional development:Dr. Ballén-Taborda and all graduate students in the CGBG program were able to attend the National Association of Plant Breeders (NAPB) annual meeting hosted by Clemson University on 16-20 July 2023 in Greenville, SC, with PD Boyles co-organizing and presenting at the event. MS graduate Behnke presented a poster at this meeting as well as at the 2023 Plant and Animal Genome Conference in San Diego, CA. Dr. Ballén-Taborda, as highlighted previously, presented project research at multiple conferences including the 2022 ASA, CSSA, SSSA International Annual Meeting and an American Seed Trade Association Conference in 2023. Project members also attended a WheatCAP genomics workshop held in Raleigh, NC following the 2023 NAPB meeting. Dr. Ballén-Taborda led a workshop on how to run genomic prediction and use the PopVar program for predicting progeny variance. Professional development opportunities for non-project funded early-career scientists:Students, international scientists, and stakeholders representing diverse ethnic groups attended a weeklong virtual workshop event called theOrganic Plant Breeding Institute, heldJune 2021. Based on reporting, 280individuals registered for one or more events, with a total of 218attendees that viewed all or part of the week-long workshop. During this event, PDBoyles (organizer and co-host) and team members at Clemson University provided (1) a detailed overview of the Clemson Cereal Grains Breeding & Genetics wheat and sorghum breeding programs, (2) individual presentations onconventional breeding methods and techniques for wheat and sorghum that included emasculation, use of cytoplasmic male sterility, and cross-pollination, and (3) live tutorials of processing raw genomic data, analyzing genetic relatedness and diversity, genetic mapping strategies, and genomic prediction models and implementation. These collective presentations provided thorough insights into how to successfully develop new crop cultivars using a combination of molecular and conventional breeding tools. PD Boyles also provided guest lectures annually to the Clemson Plant Breeding course (PES4050) on emerging tools in applied plant breeding. Project director professional development: PD Boyles benefited tremendously from this project in numerous ways. Briefly, PD Boyles expanded his research program to deploy advanced breeding approaches such as genomic prediction and doubled haploid technology. These approaches collectively make the Clemson CGBG program very competitive in developing superior winter wheat and small grains for the southeastern US to get better genetics into the hands of growers. PD Boyles was able to recruit top talent into the program, including postdoc Dr. Carolina Ballén-Taborda and MS graduate Mary-Frances Behnke. PD Boyles was able to build his collaborative network with the SunGrains cooperative program to increase research output and product development. PD Boyles was able to annually attend the NIFA Project Director's Meeting held in conjunction with the NAPB meeting to understand current and future grant opportunities and provide feedback on existing initiatives and input on future research directions. In significant part to this project, PD Boyles was awarded Junior Researcher of the Year in 2023 by the Division of Research, which recognizes the top junior faculty across the entire university. How have the results been disseminated to communities of interest?Crop producers:A new website(www.sungrains.org) was developed in late 2022 and went live in January 2023 that provides information about the SunGrains demonstration trial, named the SunShow. The SunShow, grown in five southeastern US locations, is intended to serve as a resource for seedsmen and potential licensees of new cultivar releases. Interested individuals will be able to observe each entry (wheat and oats), scan a QR code on a sign in front of each entry, and access the website for detailed information about each entry, such as important QTL information and overall performance across environments. The website has been expanded to include more information on individual SunGrains breeding programs to increase awareness and outreach. Data generated from statewide variety testing were collected, analyzed, andpublished annually on the Clemson University small grains variety testingwebsite (www.clemson.edu/cafls/research/vt/small-grain.html). A Pee Dee Research and Education Center Small Grains Field Day was held onApril 27, 2023. PD Boyles, Small Grains Extension Specialist Alex Coleman, and Extension Agent David DeWitt organized and publicized the event. Attendees of the event included wheat growers from NC and SC, seed company representatives, animal production company representatives, chemical company representatives, regional small grains breeders, and university extension agents. Speakers at the event included Robert Etheridge (President of Mixon Seed), Dr. Christina Cowger (USDA-ARS Raleigh), Alex Coleman (Clemson small grains extension specialist), Dr. Mike Marshall (Clemson weed scientist), and PD Boyles. A second wheat field day is scheduled for April 25, 2024 to update wheat producers and industry stakeholders on final project products, particularly new wheat varieties released or nearing release. PD Boyles did presented at a GrowPro Genetics industry field day in May 2022 that was hosted by grower Carl Coleman (also, owner of Little Mill Seeds) on his farm in Dillon, SC. PD Boyles was an invited speaker at the North Carolina Small Grain Growers Association annual board meeting held at Research Triangle Park (Durham, NC) in January 2023, where over 40 wheat growers were in attendance. PD Boyles was also invited to present at the first annual joint conference between the NC Seedsmen's Association and SC Seedsmen's Association in July 2023 in Charleston, SC. SunGrains breeders:Both genomic and phenotypic data collected on lines evaluated in cooperative field trials during project years were shared among SunGrains breeding programs, primarily through group email sharing of flat (CSV) files. The SunGrains group maintained a website datahub (www.sungrains.lsu.edu/data.shtml) that stored field data for every SunGrains nursery and site-year to support historical data preservation and use. The group also has established a Google folder for sharing genomic data, genomic prediction datasets, and other relevant information. A Slack channel was created to share observations and information instantaneously to other breeding programs, including students that can learn what experienced breeders are seeking as well as what new/relevant challenges are occurring in production fields. SunGrains breeders met twice each year: (1) spring field tour to observe field trials across mega-environments and (2) summer meeting to discuss prior year results and research progress while also planning upcoming research and breeding trials prior to fall planting. New training sets and genomic prediction models were shared among the group, with Dr. Ballén-Taborda presenting project research results in the winter journal club nicknamed "Plant Breeding Gumbo." Scientific Community:The project team published 15 journal articles, a book chapter, and gave 12 presentations to disseminate research to the scientific community. Another three manuscripts are in various stages of the publication process to further increase outreach. Notably, Dr. Ballén-Taborda published peer-reviewed journal articles on genomics-enabled wheat improvement strategies [Ballén-Taborda et al. (2022) Frontiers in Genetics and Ballén-Taborda et al. (2024) Crop Science (conditionally accepted)]. PhD graduate Dr. AJ Ackerman published an impactful research article on advanced phenotyping for wheat disease evaluation (Ackerman et al. 2022 Agronomy). Also, former NC State University PhD student Zach Winn published an article in Theoretical Applied Genetics about the ability of machine learning to predict major effect QTL presence/absence from genotyping-by-sequencing data. PD Boyles published a review article on the advances of wheat resistance to Fusarium head blight in the southeastern US region (Boyles et al. 2023 Plant Breeding). These published research articles are open-access and available to the broader research community. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Impact statement:This project leveraged genomics and cooperative field testing to make significant advances in the development of soft red winter wheat for the southeastern US. This region, with more than 2.5 million wheat acres, relies heavily on public breeding programs to provide superior genetics to produce safe, nutritious, and high-quality wheat for food and feed. From this project, the Clemson wheat breeding program built a reliable, accurate genomic prediction platform that is being and will continue to be used for preliminary screening of line performance of 1000s of lines annually. Genomics-enabled breeding now estimates yield and test weight while also predicts the presence or absence of key alleles for disease and pest resistance, photoperiod, and vernalization. Doubled haploid (DH) technology was leveraged to expedite line development, with one DH cultivar (SCLANC11558-33) currently going through the Clemson University release process. Many more Clemson advanced wheat lines are at final stages of SunGrains cooperative field testing that show significant promise for future commercialization.Collectively, the tools and germplasm generated from this project set up the Clemson wheat breeding program for short- and long-term success in cultivar development. Genomic selection for regional soft wheat improvement:Historical phenotype data (19 locations, 15 years) for grain yield, test weight, heading date, and plant height of 1,149 advanced wheat breeding lines were obtained from the SunGrains breeding cooperative. Genotype plus genotype-environment interaction (GxE) biplots and PCA plots were used to identify locations that correlated with Florence, SC. Genome-wide marker data consisting of 15,077 SNPs and 9,137 wheat genotypes, including the 1,149 advanced breeding lines from SunGrains and 998 Clemson preliminary breeding lines selected in South Carolina as part of the project were used to optimize training sets for forward prediction. Cross-validation was used to calculate the correlation between the observed and predicted values (i.e.GEBVs) to measure model accuracy. Focusing on grain yield, we obtained an average accuracy of 0.42 among all training populations but optimal training sets generated accuracies as high as 0.62 (Ballén-Taborda et al. 2022). The accurate models built for predictions will increase the likelihood to identify and select the most promising lines for regional testing, thus increasing the probability of selecting a superior line worthy of release and commercialization. In addition, we performed a QTL-by-environment interaction analysis to study whether the presence/absence of major effect QTLs influenced wheat productivity across three mega-environments [Region1: Atlantic Coastal Plain (NC, SC, VA), Region 2: GA and FL, and Region 3: Gulf Coast (TX, AR, LA)]. We studied 18 major effect genes associated with important pest and disease traits. We found that breeding lines harboring certain QTL (e.g.Fhb1) in the absence of disease could potentially underperform to drag down yield while others such as the Yr17 resistance allele had a positive yield impact irrespective of presence or absence of disease. This research highlighted the importance of genomics-enabled breeding and the establishment of multi-institutional breeding cooperatives to accelerate the development of new superior cultivars. This research was published in Frontiers in Genetics (Ballén-Taborda et al. 2022) and presented by Dr. Ballén-Taborda at the 2022 ASA-CSSA-SSSA meeting. Prediction of best cross-combinations that give rise to superior progeny:With the assumption that superior crosses would produce populations that would be retained and advanced by breeders, or that would likely be released, a retrospective PopVar study was performed to test the validity of genomic prediction of progeny performance traits. A total of 217 unique parents representing 668 elite breeding lines were utilized to produce 23,436 PopVar simulated progenies. When we compared the predicted and observed values of 760 breeding lines, low-to-moderate correlations were observed, 0.30 for grain yield, 0.25 for test weight, 0.52 for heading date, and 0.27 for plant height. Importantly, breeding lines that persisted longer in the breeding program tended to originate from families that were predicted to have higher grain yield, including two released cultivars. This research helps reduce manual crossing efforts and save operational costs and space by lowering the number of crosses made per cycle. This research was presented by Dr. Ballén-Taborda at the Clemson Postdoctoral Symposium, where she was awarded 3rdplace prize, as well as at the 2023 ASTA Conference. The manuscript titled "Predicting superior crosses in winter wheat using genomics: A retrospective study to assess accuracy" was just conditionally accepted in Crop Science. Empirical field studies were also completed in project years 2-3 to further validate progeny predictions by evaluating more than 2,000 plants (F2lines and parents) each year from approximately 30 biparental families. Individual plants were barcoded when collecting heading data to further measure plant height and grain yield components. This multiyear dataset provided a rich resource to empirically evaluate the ability of genomic prediction models to estimate progeny usefulness (genetic variance, population mean, transgressive segregant percentage, etc.). A manuscript highlighting this work is being finalized by Dr. Ballén-Taborda for peer-review. Incorporation of doubled haploids into cooperative field testing:In 2023, the Gulf Atlantic Wheat Nursery, the most advanced soft winter wheat nursery evaluated in the southeastern US, evaluated 10 DH lines of 54 experimental lines in total (19%). The DHs yield significantly better (DH: 74.1 bu/ac; Non-DH: 68.5 bu/ac) than the lines selected using conventional breeding practices, with three of the top seven entries being DH lines. Across all advanced, cooperative nurseries in 2022-2023, there were 58 DHs represented out of 171 total entries (34%). Of 337 first-year DHs grown in Clemson headrows, 58 were selected by PD Boyles in 2022-2023 and entered into 2023-2024 preliminary yield trials. Another 560 DHs (representing five families) were planted in headrows in November 2023 for evaluation and selection in the 2023-2024 field season. One doubled haploid (DH) line, SCLANC11558- 33, ranked 1stand 3rdfor grain yield in the Georgia statewide variety trial in 2023 and 2022, respectively. SCLANC11558-33 is going through the release and seed purification processes at Clemson University. Regional yield trials for advanced breeding line evaluation:In each project year, the SunGrains cooperative breeding program coordinated advanced nurseries in 10 states (AR, FL, GA, KY, LA, MD, NC, SC, TX, and VA) within the greater southeastern USA. Advanced trials collectively evaluated over 400 breeding lines in each season. Three Clemson University wheat lines that advanced from these trials were entered into the Uniform Southern Soft Red Winter Wheat Nursery (USSRWWN), which is grown in over 20 environments in the eastern USA. Based on yield means over locations, two of the Clemson entries in the 2022-2023 USSRWWN displayed wide adaptation. During the project, SunGrains evaluated some 1,500 advanced soft winter wheat lines while running preliminary testing on more than 10,000 lines in this 3-yr period. Preliminary testing including low replication field testing (e.g.Clemson evaluated nearly 2,000 lines in two location, single rep yield trials over three years) and genomic prediction to estimate the breeding value of these 10,000 lines, which was conducted using a historical training population and over 15,000 SNPs. This project therefore generated more than 150 million genetic datapoints and over 20,000 phenotypic datapoints, all of which are being used to accelerate wheat development.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Schoen A, Wallace S, Holbert MF, Brown-Guidera G, Harrison S, Murphy P, Sanantonio N, Van Sanford D, Boyles R, Mergoum M, Rawat N. Reducing the generation time in winter wheat cultivars using speed breeding (2023) Crop Science 63(4):2079-90. https://doi.org/10.1002/csc2.20989
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
DeWitt N, Lyerly J, Guedira M, Holland JB, Murphy JP, Ward BP, Boyles RE, Mergoum M, Babar MA, Shakiba E, Sutton R, Ibrahim A, Tiwari V, Santantonio N, Van Sanford DA, Howell K, Smith JH, Harrison SA, Brown-Guedira G (2023) Bearded or smooth? Awns improve yield when wheat experiences heat stress during grain fill in the southeastern United States. Journal of Experimental Botany 74(21):6749-59. https://doi.org/10.1093/jxb/erad318
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Boyles RE, Ball�n-Taborda C, Brown-Guedira G, Costa J, Cowger C, DeWitt N, Griffey CA, Harrison SA, Ibrahim A, Johnson J, Lyerly J, Marshall DS, Mason RE, Mergoum M, Murphy JP, Santantonio N, Saripalli G, Sutton R, Tiwari V, Van Sanford D, Winn ZJ (2024) Approaching 25 years of progress towards Fusarium head blight resistance in southern soft red winter wheat (Triticum aestivum L.). Plant Breeding 143(1):66-81. https://doi.org/10.1111/pbr.13137
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
DeWitt N, Guedira M, Lyerly J, Ward BP, Murphy JP, Marshall D, Santantonio N, Griffey CA, Boyles RE, Mergoum M, Johnson J, Harrison S, Babar MA, Mason RE, Ibrahim A, Van Sanford DA, Tiwari VK, Kuraparthy V, Kolb FL, Sneller CH, McKendry AL, Mohammadi M, Maltecca C, Tiezzi F, Brown-Guedira G (2021) Unpacking the Yield Effects of Major Heading Date Alleles in Wheat through Joint Analysis of Historic Breeding Panels and Their Climates. ASA, CSSA, SSSA International Annual Meeting, Salt Lake City, UT. 10 November 2021.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
McBreen J, Babar MA, Hernandez-Jarquin JD, Lyerly J, Mergoum M, Murphy JP, Boyles RE, Harrison S, Ibrahim A, Shakiba E, Sutton RL (2022) Improving Grain Yield in Wheat Lines Adapted to the Southeastern United States through Multi Trait and Multi Environment Genomic Prediction Models Incorporating Spectral and Thermal Information. ASA, CSSA, SSSA International Annual Meeting, Baltimore, MD. 8 November 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
DeWitt N, Lyerly J, Brown-Guedira G, Rogers A, Holland J, Winn Z, Harrison S, Santantonio N, Murphy JP, Boyles RE, Mergoum M, Shakiba E, Sutton RL, Ibrahim A, Babar MA (2023) Incorporation of Environmental Data and Developmental Phenotypes to Explicitly Model and Predict GxE in Historic Sungrains Breeding Cooperative Yield Trials. ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO. 30 October 2023.
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Babar MA, Harrison SA, Blount A, Barnett RD, Johnson J, Mergoum M, Mailhot DJ, Murphy JP, Mason RE, Ibrahim A, Sutton R, Simoneaux B, Boyles R, Stancil B, Marshall D, Fountain M, Klos KE, Khan N, Wallau M, Jordan HG (2023) FLLA09015-U1: A broadly adapted dual-purpose oat cultivar for southern USA. Journal of Plant Registrations 17(2):228-37. https://doi.org/10.1002/plr2.20249
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Babar MA, Harrison SA, Blount A, Barnett RD, Johnson J, Mergoum M, Mailhot DJ, Murphy JP, Mason RE, Ibrahim A, Sutton R, Simoneaux B, Boyles R, Stancil B, Marshall D, Fountain M, Klos KE, Khan N, Wallau M, Jordan HG, Ma X, Arbelaez J (2023) FLLA11019-8: A new dual-purpose facultative oat cultivar for grain and forage production in the southern United States. Journal of Plant Registrations 17(2):238-46. https://doi.org/10.1002/plr2.20272
- Type:
Journal Articles
Status:
Under Review
Year Published:
2024
Citation:
Ballen-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Ward B, Babar MA, Harrison S, Mason RE, Mergoum M, Murphy J, Sutton R, Griffey C, Boyles RE. Predicting superior crosses in winter wheat using genomics: A retrospective study to assess accuracy. Crop Science. Accepted.
- Type:
Journal Articles
Status:
Submitted
Year Published:
2024
Citation:
Ballen-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, Babar MA, DeWitt N, Harrison S, Mergoum M, Murphy J, Shakiba E, Sutton R, Boyles RE. Predicting superior crosses in winter wheat using genomics: A field-based empirical validation. In preparation.
- Type:
Other
Status:
Accepted
Year Published:
2023
Citation:
Oral presentation: Boyles RE (2023) Building greater resiliency in southern small grains, NC and SC Seedsmen Association, Charleston, SC. 24 July 2023.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Field day talk: Boyles R (2023) The revamped Clemson University wheat breeding program, Clemson University Pee Dee REC Small Grains Field Day, Florence, SC. 27 April 2023.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Seminar: Boyles R (2022) Harnessing diversity and leveraging genomics for cereal crop improvement, University of Nebraska, Lincoln, NE. 21 October 2022.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Babar MdA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA, Boyles RE (2023) Prediction and evaluation of parental combinations using winter wheat historical data. National Association of Plant Breeders Annual Meeting, Greenville, SC. 17 July 2023.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Babar MdA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA, Boyles RE (2023) Prediction and evaluation of parental combinations using historical data. The American Seed Trade Association (ASTA) - 62nd Vegetable & Flower Seed Conference, Orlando, FL. 28 January 2023.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Ball�n-Taborda, C, Boyles RE (2021) PopVar predictions of progeny performance and validation in wheat (Triticum aestivum). 1st Annual Postdoctoral Symposium, Clemson University Postdoctoral Association (CUPDA), Clemson, SC. 10 December 2021. Received 3rd place award in competition.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
DeWitt N, Lyerly J, Brown-Guedira G, Rogers A, Holland J, Winn Z, Harrison S, Santantonio N, Murphy JP, Boyles RE, Mergoum M, Shakiba E, Sutton RL, Ibrahim A, Babar MA. Incorporation of Environmental Data and Developmental Phenotypes to Explicitly Model and Predict GxE in Historic Sungrains Breeding Cooperative Yield Trials, ASA, CSSA, SSSA International Annual Meeting, St. Louis, MO. 30 October 2023
- Type:
Journal Articles
Status:
Submitted
Year Published:
2024
Citation:
McBreen J, Babar MA, Jarquin D, Khan N, Harrison S, DeWitt N, Mergoum M, Lopez B, Boyles R, Lyerly J, Murphy JP, Shakiba E, Sutton R, Ibrahim A, Howell K, Smith JH, Brown-Guedira G, Tiwari V, Santantonio N, Van Sanford DA. Improving grain yield in wheat lines adapted to the southeastern United States through multivariate and multi-environment genomic prediction models incorporating spectral and thermal information. Under review.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Babar MA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA, Boyles RE (2023) Identifying superior parental combinations through simulation of progeny performance in winter wheat. Virtual Plant Breeding GUMBO Journal Club. 15 February 2023.
|
Progress 01/15/22 to 01/14/23
Outputs
Target Audience:Target Audiences:In year 2 of this 3-year project, wheat industry stakeholders remain the primary audience to support through the persistentdevelopment and testing of advanced wheat cultivars.Traits such as yield and test weight that directly impact wheat producers remain highest priority, but another focal trait being Fusarium head blight resistance is targeted to increase soft wheat quality in the region that directly benefits millers and livestock producers by reducing mycotoxin levels in the wheat supply chain. Certainly, this breeding objective indirectly supports thegeneral public, rather consumers of wheat-based products and meat-based protein produced using wheat, by helping to provide safe and healthy wheat-dependent products in the regional and national marketplace. In this reporting period, project personnel delivered applied research information to the scientific community that was specifically aimed to support new or emerging breeding programs. Research results were routinely and directly shared withinstitutional breeders within the Southeastern University small grains breeding cooperative (SunGrains).Clemson University postdoctoral research fellow Dr. Carolina Ballen-Taborda (female minority) continued to participate greatly in the project as well as Clemson MS student Mary-Frances Behnke (female). Efforts:Dr. Boyles gave presentations on advanced breeding lines and likely future cultivar releases at aGrowPro Genetics wheat field day event in Dillon, SC on 3 May 2022 as well as at the NC Small Grain Growers Association annual meeting in Durham, NC on 11 January 2023. Dr. Boyles visited six different small grains farm operations in the reporting period to gain feedback oncriticalproductionchallenges that could be addressed through plant breeding and genetics. In addition, feedback was received from crop consultants and private companies that licensesmall grains releases from the Clemson program to understand how well varieties performed and if particular improvements can be made in future cultivars. In February 2022, Dr. Boyles participated in the USDA NIFA Wheat Resiliency Conference led by the University of Kentucky to share his vision to colleagues on how to effectively prepare the wheat crop for impending climate change. Dr. Boyles attended the 2022 National Fusarium Head Blight Forum in Tampa, FL to provide updates on research progress to collaborators within the Variety Development and Host Resistance (VDHR) Southern Winter Wheat (SWW) coordinated project, which is supported by the US Wheat & Barley Scab Initiative. At this meeting, Dr. Boyles also provided a regional update on wheat industry personnel and production changes to the National Wheat Improvement Committee (NWIC). Postdoc Carolina Ballen-Taborda presented project research findings at the 2022 ASA, CSSA, SSSA International Annual Meeting in Baltimore, MD, and Dr. Ballen-Taborda also placed third in a university-held poster competition held by the Clemson University Postdoctoral Association. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Early career scientists:This project has served and will continue to serve as an important platform to provide training for early career researchers on applied genomics-enabled breeding methodologies, data analysis and for professional development opportunities. One postdoc and a graduate student werehired in summer 2021 and have continued working on data management, genomic analyses, interpretation, and publication of research findings inpeer-reviewed journals. Dr. Carolina Ballén-Taborda, with a PhD in Plant Breeding, Genetics, and Genomics from the University of Georgia, has completed the implementation and optimization of genomic prediction models for the South Carolina region, by incorporating large historical genomic and phenotype data generated and maintained by SunGrains. This research was published inFrontiers in Geneticsjournal (Ballén-Tabordaet al2022) and presented at the ASA-CSSA-SSSA Crop and Soil International Annual Meeting in November 2022.Furthermore, Dr. Ballén-Taborda has worked on performing simulation of progeny performance using the PopVar tool. This has included two different research projects: 1) 2-year PopVar predictions and field validation, and 2) retrospective PopVar analysis to study whether this tool could allow breeders to develop new cultivars by focusing on promising parental combinations. Is anticipated publishing at least one article from this research. This research was presented at an on-campus venue in 2021 (the 1st Annual Postdoctoral Symposium) and will be presented in January 2023 at the American Seed Trade Association (ASTA) - 62nd Vegetable & Flower Seed Conference. Dr. Ballén-Taborda hasimproved her data analysis and programming skills in R and continue to be trained and advised directly by the PD Dr. Richard Boyles and Dr. Jeanette Lyerly from NS State. Finally, she has participated in SunGrains meetings to interact with wheat breedersand attended interactive workshops and trainings in R and Linux with the Clemson Computing and Information Technology group (CCIT). Graduate students AJ Ackerman (PhD earned in August 2022) and Mary-Frances Behnke (MS degree expected in May 2023) have been involved with most wheat cultivar development stages such as greenhouse crossing, early generation bulk field selections, headrow evaluation, data collection in yield plots, combine and headrow harvest, and post-harvest grain processing. Both have contributed to evaluating wheat for Fusarium head blight resistance, which Dr. Ackerman recently publishedon this effort (Ackerman et al. 2022 Agronomy). Dr. Ackerman successfully landed a postdoctoral research position at the University of Illinois Urbana-Champaign with the wheat breeding and genetics program. Ms. Behnke attended the 2023 Plant and Animal Genome Conference (13-18 January) to present some of her research findings. Project Director: PD Boyles attended the 2022 National Association of Plant Breeders Annual Meeting in Ames, IA from 8-11 August. At this event, PD Boyles also attended the NIFA A1141 Cultivar Development Project Director's Meeting and was able to network with renowned plant breeders and provide input on the future priority research areas for A1141 Cultivar Development. PD Boyles was able to provide four differentpresentations in project year 2 that included an invited seminar at the University of Nebraska Lincoln and an invited presentation at the New Crops for the Southeast conference held by Tennessee State University. PD Boyles published nine journal articles in 2022 that contributed to information dissemination across the scientific community. ?Research Technical Staff: Last but certainly not least, research technicians William Caughman, Dawn Hicks, andRyan Holmes were able to successfully carry out all requested cultivar development assignments in project year 2, which helped to build their practical and managementskills. This strong technical support team made it feasible to generate high-quality field data needed to make accurate breeding decisions and share information with the wheat community, including producers, seedsmen, and scientists. How have the results been disseminated to communities of interest? Crop producers: A new website (https://www.sungrains.org/) was developed in late 2022 and went live on 4 January 2023 that provides information about the SunGrains demonstration trial, named the SunShow. The SunShow, grown in five southeastern US locations,is intended to serve as a resource for seedsmen and potential licensees of new cultivar releases. Interested individuals will be able to observe each entry (14 wheat and eight oat lines), scan a QR code on a sign in front of each entry, and access the website for detailed information about each entry, such as important QTL information and overall performance across environments. Data generated from statewide variety testing and the inoculated fusarium head blight (FHB) nursery were collectedagain in spring 2022, analyzed, and will be published on the ScabSmart website(scabsmart.org/South_Carolina_SWW, www.clemson.edu/cafls/research/vt/small-grain.html). Results from the DON Testing Laboratory at Virginia Tech have not yet been received and final data will be reported on ScabSmart upon its receipt and verification.Thefield day planned for project year 2 (spring 2022)was planned but inevitably was postponed due to two reasons: (1) lingering concern around COVID-19 and (2) PD Boyles went on paternity leave at the scheduled time of the planned event (late April). The 2023 PDREC Small GrainsField Day has already been planned and scheduled for Thursday, 27 April, with PI Boyles, Small Grains Extension Specialist Alex Coleman, and Extension Agent David DeWitt coordinating to organize and publicize the event.In year 2, PD Boyles did attend and present at aMay 2022 GrowPro Genetics industry field day that was hosted by grower Carl Coleman (also, owner of Little Mill Seeds) on his farm in Dillon, SC. Finally, PD Boyles was an invited speaker at the North Carolina Small Grain Growers Association annual board meeting held at Research Triangle Park on 11 January 2023, where over 40 wheatgrowers were in attendance. SunGrains breeders: Both genomic and phenotypic data collected on lines evaluated in2021-2022 cooperative field trials were shared among SunGrains breeding programs, primarily through group email sharing of Excel files. The SunGrains group maintains a website datahub (http://www.sungrains.lsu.edu/data.shtml) that can store field data for every SunGrains nursery and site-year. The group also has established a Google folder for sharing genomic data, genomic prediction datasets, and other relevant information. A Slack channel has also been created to share observations and information instantaneously to other breeding programs, including students that can learn what experienced breeders are seeking as well aswhat new/relevantchallenges are occurring in production fields. Plant scientists: Clemson University postdoc Ballen-Taborda and PhD student Ackerman both publishedpeer-reviewed journal articles on wheat improvement strategies, focusing on genomic (Ballen-Taborda et al. 2022 Front Genet) and phenotypic evaluation (Ackerman et al. 2022 Agronomy) methods, respectively. Also, former NC State University PhD student Zach Winn published an article in Theoretical Applied Genetics about the ability of machine learning to predict major effect QTL presence/absence from genotyping-by-sequencing data. All of these original research articles published in 2022 are open-access. What do you plan to do during the next reporting period to accomplish the goals?Simulation of population variance (PopVar): For a second field validation study that was planted in winter 2022, data collection of HD,PH, and YLD components (TSN, TSW, TKW, and TKNS) will be completed in year 3. The 30 F2populations(10 each fromtop,middle, andbottom predicted categories)will be harvested and scored in the same way we did for the first validation experiment (PopVar 2021-22). Furthermore, the 2-year data comprising PopVar predictions for four agronomic traits (YLD, TW, HD and PH) and field observed data (YLD, HD, PH TSN, TSW, TKW and TKNS) will be analyzed. Mid-parent values, genetic variance (VG), population mean (??) and 10% superior/inferior progeny mean (??sp/??ip) values for each trait and each population will be contrasted for validation. If the prediction and validation data correlate (at least for yield), this will encourage breeders to use progeny simulations to help them inform and guide the decisions of the most promising crosses to make. QTLxEnvironment analysis using predicted QTL data: QTL-by-environment interaction analysis will be made to study whether the predicted presence/absence of major effect QTL influence four agronomic traits (YLD, TW, HD and PH) across three mega-environments in the southeastern USA. Here, we will be able to report the effect of presence/absence of individual QTL and the effect of combination of multiple genes. For predicting the presence/absence of a major affect genein collaboration with researchers from NC state (see Winn et al. 2022 Theor Appl Genet), and the effect will be detected using similar methods as previously described (Ballén-Tabordaet al2022). Incorporating environmental covariates into GS to improve prediction accuracy and GxE modeling:One of the main goals of breeders is to develop new cultivars adapted to a wide range of environmental conditions. However, predicting how individual breeding lines interact with a given environmentand measuring how these interactions impactyield and other agronomic traits, is challenging. In this context, we are interested in incorporating historical weather data (e.g. precipitation, temperature, soil characteristics) as covariates to account for genotype-by-Environment (GxE) interactions into existing genomic prediction models to assess if prediction accuracy could be enhanced. This will be useful to predict line performance within and across locations. Submit documentation for proposedwheat releases and Plant Variety Protection: Wheat cultivar SCLANC11558-33 was planted on five acres in November 2022 for certified seed production and further entered into regional state variety trials. If this cultivar remains competitive as in years past, this cultivar will likely be presented to the Clemson University Variety Review and Release Committee (VRRC) to request cultivar release and enable downstream licensing. Extensive phenotypic data will be collected during the 2022-2023 field season that will support application filing for Plant Variety Protection (PVP). The timeline for cultivar release would be receiving approval by the VRRC in summer 2023,filing the PVP application in fall 2023, and working with theClemson University Research Foundation to put out a Request For Bid in fall 2023 to seed companies for licensing the wheat cultivar. ?Host a small grains field day event to promote outreach on newly developed and recently released wheat cultivars:The 2022 Small Grains Field Day was unfortunately postponed due to (1) ongoing concern around the pandemic and (2) PD Boyles being on paternity leave in April 2022. However, the 2023 Pee Dee RECSmall Grains Field Day has been scheduled for 27 April. Meeting planning and publicizing are bothunderway, with organizers being PD Boyles, Pee Dee REC Director Matt Smith, Small Grains Extension Specialist Alex Coleman, and County Agent David DeWitt.
Impacts
What was accomplished under these goals?
Genomic selection forregional soft wheat improvement:In year 2, we performed a QTL-by-environment interaction analysis to study whether the presence/absence of major effect QTL (based on PCR-marker calls) have an influence on YLD, TW, HD and PH across three mega-environments in the southeastern US [Region1: Atlantic Coastal Plain (NC, SC, VA), Region 2: GA and FL locations, and Region 3: Gulf Coast states (TX, AR, LA)]. We studied 18 major effect genes associated with resistance to FHB, leaf rust, stem rust, stripe rust, hessian fly, powdery mildew, susceptibility to Septoria nodorum blotch, and underlying photoperiod sensitivity. We observed that breeding lines harboring major QTL in the absence of disease could potentially underperform (e.g. Fhb1 R-gene), whereas it is advantageous to express a major QTL under biotic pressure (e.g., stripe rust R-gene). This research highlighted the importance of genomics-enabled breeding and the establishment of muti-institutional breeding cooperatives to accelerate the development of new commercial cultivars for the target population of environments. This research was published in Frontiers in Genetics journal (Ballén-Taborda et al 2022) and presented at the ASA-CSSA-SSSA Crop and Soil International Annual Meeting in November 2022. Simulation of population variance:Using phenotypic and genotyped data of 3,084 SunGrains breeding lines, we completed PopVar (Mohammadi et al. 2015)predictions of progeny performance for four agronomic traits (YLD, TW, HD and PH) in years 1 and 2.Empirical field validation was completed in year 2 (2021-2022) and will also be performed in year 3 (2022-2023). PopVar predictions were completed in year 1 for YLD, TW, HD, and PH for 2,016 simulated progenies derived from 64 parents. Based on YLD predictions, 33 F2 populations(10 each fromtop,middle, andbottom predicted categories) were selected for first-yearfield validation. The field trial was planted in an alpha lattice design with 2 blocks for each population that included 126 F2 seeds and 16 seeds of each of the corresponding female and male parents. For 24 of the planted 33 F2 populations, 40 F2 plants, four females and four males were uniquely marked with barcoded plastic tags when collecting HD data and later PH data was measured. At harvest, spikes for eachplant for each population were collected, and four yield components were measured including total spike number (TSN), total seed weight (TSW) (g), 1000-kernel weight (TKW) (g), and total number kernels per spike (TNKS).Furthermore, data quality check and cleaning of HD,PH, and YLD components were implemented. In year 2, PopVar predictions for YLD, TW, HD and PH were performed with 68 parents to produce 2,278 progeny simulations. Based on YLD predicted values, 30 F2 populationswere selected to complete a second field validation. This was planted the same way as the field experiment in year 1. This multiyear dataset will provide a rich resource to empirically evaluate the ability of genomic prediction models to estimate progeny usefulness (genetic variance, population mean, transgressive segregant percentage, etc.). Retroactive PopVar: With the assumption that superior crosses would produce populations that would be retained and advanced by breeders, or that would likely be released, retrospective PopVar predictions and evaluation were performed. To complete this, the pedigree of 3,084 SunGrains advanced breeding lines was used to identify lines derived from two-way crosses. Based on this information, a total of 217 unique parents representing 668 elite breeding lines and 2 released cultivars (GA09436-16LE12 and AR09137UC-17-2) were utilized to produce 23,436 PopVar simulated progenies. When we compared the predicted and observed values of the 760 breeding lines, low-to-moderate correlations were observed, 0.30 for YLD, 0.25 for TW, 0.52 HD, and 0.27 PH. These values for YLD, TW, and PH are not surprising considering that these are very complex traits, and HD is controlled by fewer genes; therefore, the observed correlation was higher. Furthermore, we studied whether these 670 lines were retained by SunGrains breeders and tested in advanced nurseries (USW, GAWN, and SunWheat)or were dropped after preliminary testing (SunPre nurseries). For yield, we observed that lines that persisted longer in the breeding progress were predicted to have high yield, including the two released cultivars. For instance, 100% of the lines advanced into USW (most advanced nursery)produced superior yield; and ~75% of the lines tested in either GAWN or SunWheat nurseries showed the same trend. Lines that did not passthe preliminary testing (SPE and SPL) had a wide range of predicted yield values. This research intendedto reduce manual crossing effortsand save operational costs and space when advancing them in the breeding pipeline. The use of predictions to enhanceefficiency of wheat breedingcould allow breeders and potentiallyacceleratethe deployment of new wheat cultivars in the marketplace.This research will bepresented at the 1st Annual Postdoctoral Symposium at Clemson University (year 1) and will be presented in January 2023 at the American Seed Trade Association (ASTA) - 62nd Vegetable & Flower Seed Conference (year 3). Incorporation of doubled haploids into cooperative field testing:The selected doubled haploid (DH) line SCLANC11558-33 that was originally developed at NC State University, subsequently evaluated by the LSU breeder, and later tested by PI Boyles performed exceptionally well in the 2021-2022 Georgia statewide variety testing program. Across several locations throughout GA, SCLANC11558-33 ranked 3rd of 78 entries for grain yield. Foundation seed of this line was also produced 2021-2022 and used for a five acre planting in fall 2022 for certified seed. This line was again distributed across the region for state variety testing. In the advanced, cooperative 2022-2023 field trial, there were 58 DHs incorporated from prior selection out of total of 171 entries (34%). These entries are being grown in six locations across the southeastern US: Florence, SC; Marianna, AR; Quincy, FL; Plains, GA; Winnsboro, LA; and Kinston, NC. Further, another 58 DHs were selected from headrows and entered into 2022-2023 preliminary yield trials that contain a total of 892 distinct lines that will undergo first-year yield evaluation. First-year doubled haploids (DHs) developed by Heartland Plant Innovations (Manhattan, KS) were planted in November 2022 for evaluation of agronomic and disease traits. There were 337 DHs planted in total that represented three pedigrees (133, 88, and 116). In addition, seed from another five F1 crosses developed in March 2022 were shipped to Heartland for doubled haploid (DH) production. A total of 200 DH lines per population (n=1,000 total) were requested for planting in project year 3 (fall 2023). ?Regional yield trials for advanced breeding line evaluation:The SunGrains cooperative breeding program coordinated advanced nurseries in 10 states (AR, FL, GA, KY, LA, MD,NC, SC, TX, VA) within the greater southeastern USA. Advanced trials, specifically the Gulf Atlantic Wheat Nursery (62 entries), SunWheat (95 entries), and SunPre (250 entries) collectively evaluated over 400 breeding lines across these 10 states in the 2021-2022 winter field season. Three Clemson University wheat lines that advanced from these trials were entered into the 2022-2023 Uniform Southern Soft Red Winter Wheat Nursery, which gets evaluated annually in over 20 environments in the eastern USA. Results from this 2022-2023 field season will largely determine whether to release one or more of these potential cultivars.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, Babar MA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA and Boyles RE (2022) Utilizing genomics and historical data to optimize gene pools for new breeding programs: A case study in winter wheat. Front Genet 13:964684. doi:10.3389/fgene.2022.964684
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Ackerman AJ, Holmes R, Gaskins E, Jordan KE, Hicks DS, Fitzgerald J, Griffey CA, Mason RE, Harrison SA, Murphy JP, Cowger C, Boyles RE (2022) Evaluation of Methods for Measuring Fusarium-Damaged Kernels of Wheat. Agronomy 12(2):532. https://doi.org/10.3390/agronomy12020532
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Ballen-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, Babar MA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton RL, Griffey CA, Boyles RE. Leveraging Genomics and Historical Data to Accelerate Wheat Cultivar Development in South Carolina. 2022 ASA, CSSA, SSSA International Annual Meeting, Baltimore, MD. 7 November 2022.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Winn ZJ, Lyerly J, Ward B, et al. (2022) Profiling of Fusarium head blight resistance QTL haplotypes through molecular markers, genotyping-by-sequencing, and machine learning. Theor Appl Genet 135:31773194. https://doi.org/10.1007/s00122-022-04178-w
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Mergoum M, Johnson J, Buck J, Buntin D, Sutton S, Lopez B, Mailhot D, Chen Z, Bland D, Harrison S, Murphy JP, Mason RE, Sutton R, Babar A, Brown-Guedira, Ibrahim A, Boyles R, Baik B-K, Marshall D, Griffey C, Cambron S, Chen X, Cowger C (2022) A new soft red winter wheat cultivar GA 08535-15LE29 adapted to Georgia and the U.S. southeast region. J Plant Registr 16:597-605. doi:10.1002/plr2.20235
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Boyles RE. Progress update from the Clemson University Cereal Grains Breeding & Genetics program. GrowPro Genetics Annual Wheat Field Day, GrowPro Genetics, Little Mill Seed, Dillon, SC. 3 May 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Boyles R. Small grains breeding in the Southeast, New Crops for the Southeast, Tennessee State University, Nashville, TN. 17 March 2022.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2023
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Babar MA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA, Boyles RE. Identifying superior parental combinations through simulation of progeny performance in winter wheat. In preparation.
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2023
Citation:
Ball�n-Taborda C, Lyerly J, Smith J, Howell K, Brown-Guedira G, DeWitt N, Babar MA, Harrison SA, Mason RE, Mergoum M, Murphy JP, Sutton R, Griffey CA, Boyles RE. Prediction and evaluation of parental combinations using historical data. The American Seed Trade Association (ASTA) - 62nd Vegetable & Flower Seed Conference. Orlando, Fl. 28 January 2023
- Type:
Websites
Status:
Other
Year Published:
2023
Citation:
Lyerly et al. (4 January 2023) Welcome to SunShow 2023. SunGrains (via SquareSpace). https://www.sungrains.org/
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2023
Citation:
Boyles RE. Spanning state lines and land-grants to enrich small grains cultivar development. North Carolina Small Grain Growers Association. Durham, NC. 11 January 2023
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Progress 01/15/21 to 01/14/22
Outputs
Target Audience:Target audiences: During this reporting period, this project targeted national and international students at the undergraduate and graduate levels that have an interest in applied breeding and genetics, cereal grains seed industry representatives that actively develop improved crop products for farmers, crop producers, and academic faculty that have current or had previous research programs that involve plant breeding, crop genetics, and/or general plant sciences. There was a focus in project year 1 to interact with collaborating scientists and communicate project deliverables. A postdoctoral research fellow (Dr. Carolina Ballen-Taborda; female minority) was hired in year 1 (June 2021) that has a background in plant genetics, genetic mapping, trait introgression, and bioinformatics. A PhD student (Yeter Karakoc; female minority) was onboarded in May 2021 to support project objectives. Efforts: (1) A plant breeding workshop was hosted by Clemson University (PI Boyles was a co-organizer and co-host) virtually in June 2021 that was free to register and attend. This effort facilitated more than 200 individuals to attend and learn about conventional and molecular breeding practices across numerous important field crops. This four day event contained presentations from national and international renowned scientists in breeding,genetics, biofortification, and human nutrition.(2) A smallspring nursery tour was hosted by PI Boyles at the Pee Dee Research and Education Center on 29-30 April 2021. Attendees were Dr. Paul Murphy (Professor, NC State), Dr. Robert Etheridge (President, Mixon Seed), Jimmy Clements (President, AgSouth Genetics), and Blake Shephard (Mixon Seed).(3) To accommodate safe practices during the COVID-19 pandemic, individual meetings with seed industry representatives were facilitated by PI Boyles during spring 2021 that enabled representatives to visit the wheat field site, observe advanced breeding lines, and discuss line performance with PI Boyles. Companies that had representatives visit included Gerad Seed Co, JoMar, KWS Cereals, Limagrain Cereal Seeds, and NorthStar. (4) PI Boyles attended a field day hosted by Mr. Carl Coleman (wheat producerand President of Little Mill Seed) on behalf ofGrowPro Genetics (formerly Syngenta Apri Pro) in Dillon, SC on 11 May 2021 to meet with various extension agents, wheat producers, and industry representatives. Changes/Problems:COVID-19 delayed thehiring of postdoc Dr. Carolina Ballen-Taborda (15 June 2021) approximately six months and graduate student (16 August 2021) approximately nine months from the projectstart date. Both scientists were expected to starton or nearer the project start date of 15 January 2021. This will result in a delay in expenditures but should not significantly impede the success and desired outcomes of the project goal and objectives. What opportunities for training and professional development has the project provided?This project has served and will continue to serve as an important platform to provide training for students and postdocs on applied genomics-enabled breeding methodologies and for professional development opportunities. One postdoc and one PhD student were hired in summer 2021 and are directly involved in data compilation, analysis and interpretation. Postdoc: Dr. Carolina Ballen-Taborda received her PhD in Plant Breeding, Genetics and Genomics from the University of Georgia in 2019. At Clemson, she has focused on the optimization of the existing genomic prediction pipeline developed by SunGrains, and simulation of population variance by targeting the South Carolina region. Part of her research was presented in an on-campus venue, the 1st Annual Postdoctoral Symposium and received the 3rd place award for the poster presentation. She also has participated in SunGrains meetings and interacted with wheat breeders from other universities in the region (NC State University, Louisiana State University, Texas A&M University, University of Arkansas, University of Florida and University of Georgia). Although Dr.Ballen-Taborda is proficient in data analysis, she did not have experience in genomic prediction workflow; therefore, she has been trained and advised directly by the PD Dr. Richard Boyles and Dr. Jeanette Lyerly from NS State. In this way, she has had the opportunity to improve her data analysis and programming skills in R. Finally, she has participated in workshops and trainings in R and Linux with the Clemson Computing and Information Technology group (CCIT). PhD student: Yeter Karakoc graduated with a MSin Plant Science, Molecular Seed Biology and Genetics from South Dakota State University in 2020. She is currently a first-year PhD student that has worked on her coursework (Bioinformatics, Principles in Field Crops, and Mycology) and developedher dissertation proposal, in which existing genomics prediction models will incorporate historical weather data to account for genotype-by-environment interaction (GxE). This effort aimsto increase the prediction accuracy of existing genomic prediction models and to be able to select breeding lines adapted to specific environments and with superior performance and quality. Her research proposal has been presented to her graduate student advisory committee that includes both PDBoyles and Dr. Ballen-Taborda. Finally, the postdoc and the student have been encouraged to explore and pursue research projects of their interest, that align with the objectives of the Clemson University Small Grains Breeding and Genetics Program. How have the results been disseminated to communities of interest?Crop producers: Data generated from statewide variety testing and the inoculated fusarium head blight (FHB) nursery were collected in Spring 2021, analyzed, and disseminated through uploading data reports online using various outlets (scabsmart.org/South_Carolina_SWW,www.clemson.edu/cafls/research/vt/small-grain.html). Unfortunately, no field day was possible in project year 1 due to the COVID-19 pandemic. However, PDBoyles did host numerous seed industry representatives (individual meetings throughout the spring) to provide an opportunity for seedsmen to view advanced wheat lines that have been released or are nearing release. Companies included AgSouth Genetics, Gerard Seed Co, KWS Cereals, Limagrain Cereal Seeds, and Mixon Seed. A Clemson University small grains field day is being planned for spring 2022. Postdoctoral training: Dr. Ballen-Taborda was educated in year 1 on genomic prediction strategies that are currently implemented in Dr. Boyles' Cereal Grains Breeding & Genetics (CGBG) program. She successfully learned about the datasets and bioinformatics pipeline that are used to facilitate generating genomic estimated breeding values (GEBVs) for individual wheat lines at the preliminary testing stage. In addition, Dr. Ballen-Taborda was able to acquire hands-on learning by supporting seed processing, preparing wheat for planting, and participate in planting in Fall 2021. Dr. Ballen-Taborda has been involved in bi-weekly CGBG program meetings that provide an environment for discussing existing program initiatives as well as brainstorming about new ideas and methodologies to improve upon the cultivar development process and accelerate genetic gain. Graduate student training:A PhD student (Yeter Karakoc; female minority) was onboarded in May 2021 and began supporting day-to-day breeding tasks that included wheat selection and harvest. Ms. Karakoc began taking coursework in Fall 2021 to satisfy her PhD requirements but has developed a dissertation project that focuses on understanding and modeling genotype-by-environments interactions (GEI) in wheat and how incorporating GEI can improve genomic prediction accuracy. She successfully formed for graduate student advisory committee and recently presented her dissertation outline to the committee in January 2022. Clemson OPBI workshop: Students, international scientists, and stakeholders representing diverse ethnic groups attended a weekly workshop event called theOrganic Plant Breeding Institute, held 7-10 June 2021, which was primarily supported by NIFA OREI project #2018-51300-28431. Based on reporting, 280individuals registered for one or more events, with a total of 218attendees that viewed all or part of the week-long workshop. During this event, PDBoyles (organizer and co-host) and team members at Clemson University provided (1) a detailed overview of the Clemson Cereal Grains Breeding & Genetics wheat and sorghum breeding programs, (2) individual presentations onconventional breeding methods and techniques for wheat and sorghum that included emasculation, use of cytoplasmic male sterility, and cross-pollination, and (3) live tutorials of processing raw genomic data, analyzing genetic relatedness and diversity, genetic mapping strategies, and genomic prediction models and implementation. These collective presentations provided thorough insights into how to successfully develop new crop cultivars using a combination of molecular and conventional breeding tools. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period we will incorporate new genotypic and phenotypic data collected between 2021 and 2022, to retrain the genomic prediction models and continue advancing and selecting superior genotypes. This also will include the simulation of progeny performance from new parental lines to allocate more crossing efforts on the best parental combinations. Correlation of predictions and field validation will allow us to suggest that genomics-enable breeding will help speed up and increase efficiency of the breeding process and maximize the genetic gain within our breeding program. Field validation of genomic predictions: Genomic prediction accuracy is key for the successful application of GS in breeding programs. We obtained a high prediction accuracy from our cross-validation analyses for yield, and to validate these results, field evaluation of 125 SC breeding lines (105 superior and 20 low-performing) is on progress in Florence, SC. Data for grain yield, test weight, heading date, plant height, disease/pest resistance (fusarium head blight, powedery mildew, leaf rust, and Hessian fly) will be collected in spring 2022, and will be contrasted with predicted GEBVs. We expect to obtain a high correlation between GEBVs and true breeding values (observed phenotypes) of individuals from the validation set. Furthermore, Prediction analysis and field validation will be submitted for publication during the next reporting year. Field validation of PopVar simulations: From each of the 33 F2populations (~100 plants each), female and male parental lines (16 plants each) planted in the field, single-plant measurements of grain yield (GY), yield components (KNP-kernel number per spike, KWS-kernel weight per spike, TKW-thousand kernel weight), test weight (TW), heading date (HD) and plant height (PH) (Lozada et al. 2019) will be recorded in spring 2022. This data will be used to calculate genetic variance (VG), mid-parent values, population mean (??) and 10% superior/inferior progeny means (??sp). Observed and predicted data will be contrasted for validation. If the prediction and validation data correlate (at least for yield), this will encourage breeders to incorporate predicted data to help them inform and guide the decisions of the most valuable crosses to make for any trait of interest. Finally, the field observations and prediction analysis will be submitted for publication during the next reporting year. QTL-by-Environment interactions (QEI) analysis: The aim is to study whether there is any effect on yield when individual loci (QTL) expression differs under different environmental pressure (e.g low or high disease pressure) across geographic space (Lowry et al. 2019). We have a compiled dataset of 4,374 breeding lines, phenotypic and genotypic data, and the information of the presence/absence of 54 QTL associated with different traits (e.g. resistance to fusarium head blight, powdery mildew, leaf rust, hessian fly, yellow dwarf virus, between others). Literature review for methods and analysis is on progress, to accurately address the analyses to perform. ?References: Lozada DN, Mason RE, Sarinelli JM, Brown-Guedira G (2019) Accuracy of genomic selection for grain yield and agronomic traits in soft red winter wheat. BMC Genetics 20:82 Lowry DB, Lovell JT, Zhang L, Bonnette J, Fay PA, Mitchell RB, Lloyd-Reilley J, Boe AR, Wu Y, Rouquette FM, Wynia RL, Weng X, Behrman KD, Healey A, Barry K, Lipzen A, Bauer D, Sharma A, Jenkins J, Schmutz J, Fritschi FB, Juenger TE (2019) QTL × environment interactions underlie adaptive divergence in switchgrass across a large latitudinal gradient. Proceedings of the National Academy of Sciences 116:12933-12941
Impacts
What was accomplished under these goals?
For the proposed objectives 1 and 3, two main research activities have been completed for the current reporting period. 1. Genomic selection for South Carolina and the southeastern USA region: Genomic selection (GS) is a methodology that uses genome-wide genotypic information, combined with phenotypic data to predict breeding values (or phenotypes) of a given trait for a set of lines of interest (Voss-Fels et al.2019). Integrating this methodology into the cultivar development pipeline can save costs and allow more resources to be devoted towards advance field testing. In this way by predicting and selecting best performing lines before testing, breeders could decrease the number of lines and increase the number of reps-locations-years for field evaluation. Having this in mind, the aim was to implement and optimize the genomic prediction pipeline to support the wheat cultivar development within the Clemson University Small Grains Breeding and Genetics Program. To accomplish this, several steps have been completed. 1.1. Training population subsetting and cross validation: First, historical data for grain yield (GY), test weight (TW), heading date (HD) and plant height (PH) of 1149 advance breeding lines were obtained from the SunGrains' breeding cooperative. It comprises data from two elite regional nurseries across 19 locations throughout the soft winter wheat region and 12years (2008-2020). Using this multi-environment dataset, genotype plus genotype-vs-environment interaction (GxE) biplots and PCA plots were obtained for yield. These plots allowed us to identify 22 subsets of locations where genotypes had similar yield to the lines grown in Florence, SC. For each of these subsets of locations, best possible estimated/predicted values were calculated using mixed (emmeans) and random (BLUPs) linear models in R, a software environment for running scripts to analyze, interpret, and present research results (https://www.r-project.org/). Second, the historical genome-wide genotypic data consisting of 15077 SNPs and 9137 entries, including the 1149 advance breeding lines from SunGrains and 998 breeding lines selected in South Carolina, was used to select sets of 400 lines (or training populations) for each of the 22 subsets mentioned above. The best training sets of 400 individuals were selected based on the genetic relatedness to the candidate/prediction set, the 998 SC breeding lines. This analysis was carried out with the STPGA package in R. Third, using the calculated emmeans and BLUP values and the SNP data for each training set, the genomic prediction mixed models were fitted using the rrBLUP package in R, and genomic estimated breeding values (GEBVs) were calculated. Finally, two types of cross validation were implemented to assess and compare each model's predictive ability or accuracy of precision: 1) Five-fold cross validation is a procedure that consists in randomly dividing the training set into 5 groups approximately equal size. Next step is masking one group, fit the model with the remaining 4 folds and predict phenotypes for the validation set. The prediction task is repeated using one-fold at a time and calculating the correlation between the observed and predicted values. Accuracy of prediction is measured as the mean of all correlations. Focusing on grain yield, we obtained an average accuracy of 0.42 between all training populations, and four of them having between 0.50 and 0.55. The highest accuracies were observed when the emmeans values of the training populations were used for prediction. 2) For cross validation, we used the grain yield data and genotypic information of 58 advance breeding lines tested in 2021. We calculated the means, emmeans and blups from the dataset, and means only for Florence, SC and its nearest representative trial location Kinston, NC. From this analysis, a prediction accuracy as high as 0.62 was obtained when the training population included all historical data from all advance nurseries, locations and years (GawnSunSetAll). Here we also observed that predictions using on emmeans had a greatest impact on accuracy. Previous studies have shown that prediction accuracy for grain yield ranges from 0.14 and 0.43 (Lozada et al. 2019). In our two scenarios- cross-validations we observed a high accuracy when using training populations that included all historical data. Predictions with high accuracy, will increase the likelihood to identify and select most-promising lines for regional testing stage, that will be rigorously tested for yield, agronomic traits and screened for disease resistance. 1.2 Execution of genomic prediction and field validation: Using the emmeans of the GawnSunSetAll training population, we completed the predictions for grain yield, test weight, heading date and plant height for the prediction set of 998 breeding lines developed at Clemson University. Based on predictions on grain yield, we selected 105 superior and 20 low-ranked lines for field testing. The field trial, that is in progress at the Clemson Pee Dee Research and Education Center (Florence, SC), was planted as an alpha lattice design with three replications, five blocks, and five standard checks per block. 1.3 Simulation of population variance (PopVar): In plant breeding, the selection of parental combinations is critical for developing superior progeny, and to maintain genetic gain over time for a given trait. Although breeders usually focus on crosses among the highest performing (elite) breeding lines to ensure the development of desired progeny, many crosses are not carried forward or are discarded in subsequent years, as they do not deliver superior progeny. Genomic selection has mostly been used to select lines to be advanced in breeding schemes or to be used as parents. Using the GS approach, the R package PopVar (Mohammadi et al. 2015) was developed to identify best crosses, by predicting phenotype (GEBVs) of progeny for all possible parental combinations. The program also predicts genetic variance (VG), mid-parent GEBVs, population mean (??) and 10% superior/inferior progeny means (??sp) for each simulated progeny. The aims of the second study for this proposal were: 1) to predict grain yield for 2016 simulated progenies derived from 64 SC elite parents using PopVar in a half diallel design [(P*(P-1))/2; P = number of parents]; and 2) to validate predictions of selected F2 populations in a field experiment. From the 2016 predicted offspring, we selected 33 F2 populations that we had availability of seed. To cover a full range of yielding progeny, 10 top, 13 middle and 10 bottom performing populations were included. The field trial for validation, that is on progress in Florence, SC., was planted with 18 x 4 blocks (630 ft deep and 26 ft wide total). Each of the 33 F2 populations were planted into two blocks, with 126 F2 seeds and 16 seeds of the corresponding parental lines. 2. Incorporation of doubled haploids into cooperative field testing: For objective 2, 20 seed of two F1 crosses developed in February 2021 were shipped to Heartland Plant Innovations (Manhattan, KS) for doubled haploid (DH) production. An estimated 200 DH lines per populations (n=400 total) are expected for planting in project year 2 (fall 2022). Selected lines will be entered into advanced regional testing for potential cultivar release. In addition, of approximately 150 Clemson wheat DH lines that were evaluated in project year 1, 27 DHs were entered into the Clemson wheat preliminary yield trial (WPT) for yield evaluation in two SC locations: Florence and Pendleton. Another 39 DH lines were selected by PI Boyles that were developed by other SunGrains institutions and are also being evaluated in the 2021-2022 WPT.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Mergoum M, Johnson J, Buck J, Sutton S, Lopez B, Bland D, Chen Z, Buntin G, Mailhot D, Babar MA, Mason R, Harrison S, Murphy JP, Ibrahim A, Sutton R, Simoneaux B, Griffey C, Bockelman H, Baik B-K, Marshall D, Cowger C, Brown-Guedira G, Kolmer J, Jin Y, Chen X, Cambron S, Boyles R (2021) A new soft red winter wheat cultivar,GA 07353?14E19, adapted to Georgia and the US Southeast environments. Journal of Plant Registrations 15:337344 doi:10.1002/plr2.20113
- Type:
Book Chapters
Status:
Published
Year Published:
2021
Citation:
Mason E, Roberts T, Boyles R, Acuna A, Arguello M, Ballesteros D, Subramanian N (2021) Chapter 6: Molecular breeding for improving waterlogging resistance in wheat. In: Alam M, Hossain M, Henry R, Seneweera S, Rakshit S (Eds.) Molecular breeding in wheat, maize, and sorghum: Strategies for improving abiotic stress tolerance and yield. Boston, MA: CAB International Publishing
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Mergoum M, Johnson J, Buck J, Sutton S, Lopez B, Bland D, Chen Z, Buntin G, Mailhot D, Babar MA, Mason E, Harrison S, Murphy JP, Ibrahim A, Sutton R, Brown-Guedira G, Simoneaux B, Bockelman H, Baik B-K, Marshall D, Cowger C, Kolmer J, Jin Y, Chen X, Cambron S, Boyles R. (2021) Soft Red Winter Wheat, GA 051207-14E53: Adapted Cultivar to Georgia and the USA Southeast Region. J Plant Registr 15:132-139. doi:10.1002/plr2.20102
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Ibrahim A, Sutton R, Johnson J, Mergoum M, Simoneaux B, Harrison S, Murphy J, Mason R, Babar A, Neely C, Opena G, Jin Y, Kolmer J, Boyles R, Cambron S, Baik B-K, Brown-Guedira G, Marshall D, Fountain M. (2021) Registration of GA06343-13E2 (TX-EL2) soft red winter wheat. J Plant Registr 15:107-112. doi:10.1002/plr2.20031
- Type:
Journal Articles
Status:
Other
Year Published:
2021
Citation:
Ballen-Taborda C, Lyerly J, Brown-Guedira G, Babar A, Harrison S, Mason R, Mergoum M, Murphy J, Sutton R, Boyles R. Utilizing genomics and historical data to optimize gene pools for new breeding programs: A case study in winter wheat (In preparation)
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