Source: IOWA STATE UNIVERSITY submitted to
ACCELERATING CORN ELITE SELECTIONS (ACES) ORGANIC BREEDING PROGRAM: NOVEL STRATEGIES TO DEVELOP FIELD & SWEET CORN FOR ORGANIC PRODUCERS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
1013072
Grant No.
2017-51300-26811
Project No.
IOW05510
Proposal No.
2017-02493
Multistate No.
(N/A)
Program Code
113.A
Project Start Date
Sep 1, 2017
Project End Date
Aug 31, 2022
Grant Year
2017
Project Director
Lubberstedt, T.
Recipient Organization
IOWA STATE UNIVERSITY
2229 Lincoln Way
AMES,IA 50011
Performing Department
Agronomy
Non Technical Summary
Our project advances goals one and eight of the USDA OREI Program: 1. Facilitating the development of organic agriculture production, breeding, and processing methods. 8. Developing new and improved seed varieties that are particularly suited for organic agriculture.Moreover, the project addresses priority area (4) for FY2017: Strengthen organic crop seed systems, including seed and transplant production and protection, and plant breeding for organic production, with an emphasis on publically available releases. Goals include: disease, weed, and pest resistance, stress tolerance, nutrient efficiency, performance in soil-improving and climate-friendly systems such as organic no-till, quality and yield improvement, and genetic mechanisms to prevent inadvertent introduction of GMO traits through cross-pollination.Our project will develop close-to-variety (elite) organic sweet and field corn genotypes carrying a genetic mechanism for spontaneous haploid genome doubling (SHGD), as well as GCI, which will enable more rapid development of superior organic varieties protected against transgene contamination. These materials will be made available to organic breeders, seed producers, and farmers. Both genetic mechanisms together will allow implementation of DH technology in organic corn breeding. The DH technology to be implemented will not require application of chemicals (e.g., colchicine), and it will be combined with gametophytic cross-incompatibility (GCI) to exclude pollen from transgenic field corn - an urgent problem for organic farmers today.Cooperative research will flourish through a network of organic farmers, government agency staff (NRCS, ARS), organic seed companies, and university researchers, based on on-station and on-farm that will include plant breeding activities and organic seed production.Extension activities, including field days, conferences, workshops, and state advisory teams, will facilitate variety evaluation, build plant-breeding capacity among organic farmers, and advance the development of superior genetic material. Organic seed companies will be full partners by participating on advisory teams, evaluating elite genotypes, and assisting in disseminating project results.A Certificate program in Organic Plant Breeding will be available to graduate students, producers, and other interested persons/parties. Classes will be offered online to allow participation around the globe.
Animal Health Component
0%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
20101201081100%
Knowledge Area
201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
0120 - Land;

Field Of Science
1081 - Breeding;
Goals / Objectives
The overall goal of the project is to substantially increase the productivity of organic field and sweet corn production within organic crop rotations, by improving the efficiency of breeding through doubled haploid (DH) technology and traditional breeding methods, focused on early plant vigor and competitiveness factors. Another goal is to develop a Certificate program in Organic Plant Breeding, which can be accessed online in the U.S. and internationally.Objectives...1. Develop novel organic field corn and sweet corn genotypes for the commercialization pipeline that utilize more efficient methods of breeding (e.g., organic-compliant Doubled Haploid technology)2. Select and test on-farm genotypes that meet the critical needs of organic producers, including high yields, competiveness with weeds, and compatibility with cover crops and ability to exclude GMO pollen3. Practice technology transfer through a network with organic farmers and organic seed companies that allows for plant-breeding, selection, evaluation and seed processing training and skill development4. Develop a graduate Certificate program in Organic Plant Breeding
Project Methods
Objective 1. Novel organic field &sweet corn genotypes for commercialization pipeline utilizing DH technology&GMO incompatibility. This project includes two thrusts for developing high performing organic field and sweet corn materials, which contain the two genetic mechanisms of spontaneous haploid genome doubling (SHGD) and gametophytic cross-incompatibility (GCI). In the first thrust, field and sweet corn DH lines already containing SHGD will be evaluated under organic field conditions, and the best lines converted for GCI. In the second thrust, a donor genotype carrying both SHGD and GCI will be developed prior to project start, elite organic sweet and field corn materials will be converted for both traits, and subsequently evaluated for testcross (TC) performance under organic conditions. In both of these thrusts, we will develop several families of lines that perform well in hybrid combinations. This will be accomplished by starting with sweet and field corn breeding germplasm with well-characterized combining ability. This will allow us to produce competitive hybrids using our new inbreds as parents. Finally, advanced organic hybrids will be evaluated under organic conditions, and educational materials and activities developed to enhance producer knowledge and dissemination of project outputs.Objective 2: Select and test on-farm genotypes that meet the critical needs of organic producers. In this component, on-farm trials in Years 2-4 will be established to conduct extensive organic evaluation of 1) the most promising field and sweet corn hybrids made from experimental inbreds developed at ISU and UW, and from the OREI corn quality breeding project; and 2) testcrosses that carry the SHGD/GCI background, when available for evaluation. Their performance in the presence of cover crops will constitute a main focus, along with their competitiveness with weeds. Once advanced lines are developed on-station, seed will be distributed to cooperating farmer-cooperators for on-farm testing and evaluation. All farms are certified organic with a history of organic corn production, and an Organic System Plan that is based on a soil-building crop rotation. Ideally, the corn crop will follow a legume and soil samples in the Fall prior to planting will determine the amount of manure application in the spring. Because the trials will be based on 'farmer reality,' farmers may adapt their management to match their farm, but manure applications will be monitored to supply no more than 150 lb N/acre to evaluate corn production across equivalent soil fertility conditions. Breeders will provide sufficient seed in order to plant three replications of 4 to 10 hybrids at each of five sites across Iowa and Wisconsin. Plot size will vary, based on land availability, but a minimum of 30 ft. x 10 ft. will be requested for each replicated plot. The strip-trial design is popular with Midwest farmer-cooperators and has shown great promise for variety evaluation. Farmer's equipment will be for used for planting and weed management, with all field operations documented in log books provided by the project.Objective 3. Practice technology transfer. Technology transfer will be an integral part of the project. Farmer-researchers will participate in the organization and implementation of annual field days and workshops in each state to advance knowledge and skills related to plant breeding and selection.Objective 4. Develop a graduate Certificate program in Organic Plant Breeding. We will develop a graduate 3-credit course that will teach both theoretical and practical aspects of Organic Plant Breeding.This course will be embedded in a 12-credit certificate program for Organic Plant Breeding.EVALUATIONThe evaluation plan for this project will include an assessment of the development and implementation of improved corn germplasm for organic systems in two states, as well as the outreach efforts associated with this project. Evaluation of the overall success of these systems will be achieved through the collection of statistical data gathered throughout this 4-year project. Analyses of the experiment will be completed using SAS software. Research findings will be presented in peer-reviewed journal articles and in Extension publications, as well as at meetings widely attended by organic stakeholders.The second part of the evaluation plan will be in the form of "plant breeding and selection" and " cover cropimplementation in corn systems" knowledge gain of cooperating farmers. Evaluation will rely on surveys distributed at field days and through mailings. Changes in participant's skills; If and how new skills and new knowledge are used; If attitudes and behaviors related to organic plant breeding have changed; and extent to which farmer participants felt the research and outreach addressed and integrated their concerns and feedback. Short-term outcomes will be measured by surveys distributed after field days and workshops. Baseline questions will be asked regarding the participants' current level of understanding of organic plant breeding, cover crops, and their value. Learning outcome questions will be included to cover the information presented at the field days and workshops. Medium term outcomes will be measured through follow up interviews with field day and workshop participants. These techniques will engage farmers in a dialogue aimed at garnering their feedback to ensure the research is relevant and applicable to their situations as well as gaining a holistic understanding of the potential barriers they face in planting more diverse germplasm in their particular organic operations.Qualitative techniques, such as interviews, are integral to understanding how farmers interpret and implement best management practices. Data concerning indicators and inputs (types of crops planted, rotation cycles, cover crop practices, interest in new lines/varieties, etc.) will be gathered at annual advisory committee meetings to further interpret the short-, medium- and long-term impacts of the project.Overall project success will be measured as 1) The number of promising field and sweet corn lines or hybrids available for organic farmers in our respective states in the 3 years following the project; 2) The number of farmers adopting recommended practices deriving from this research; and 3) The number of seed companies offering organic corn varieties deriving from this research. We also will survey for this information in our annual surveys of organic farmers in our states.

Progress 09/01/21 to 08/31/22

Outputs
Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the totality of this project, these opportunities were provided: Lubberstedt: Ph.D. students Yu-Ru Chen and Mercy Fakude (Fulbright fellow) contributed to this project. Yu-Ru Chen gave poster presentations at the RF Baker symposium in Ames, IA, 2021, and at the virtual 2021 NAPB meeting. • Scott: Trained Iowa State University Ph.D. Students Amruta Bapat and Nicholas Boerman. Trained undergraduate laboratory assistant Hannah Clubb. • Tracy: MS student Cathleen McClusky finished her MS program, and continues as Ph.D. student with Dr. Tracy. • Delate: Henry Franzen, undergraduate student, was trained in on-farm experimental protocols and data analysis for this project. How have the results been disseminated to communities of interest?Lubberstedt (lecture): • Lubberstedt, T. Past, present and future of maize doubled haploid technology. Zemun Polje conference, Belgrade Serbia (virtual) June 8, 2021. • Poster presentations of group members (see above). Scott: • Information was disseminated in the following Lectures: • Scott, M.P. Using Gametophytic Incompatibility Systems to Improve Genetic Purity of Specialty Maize. Zemun Polje conference, Belgrade Serbia (virtual) June 8, 2021. • Scott, M.P. Breeding corn for organic production systems. Iowa State University Plant Breeding Seminar, May 5, 2021. • Scott, M.P. Cross incompatibility in Corn. Iowa State University Plant Biology Faculty Seminar. September 29, 2021. • Information was disseminated through the following posters at virtual scientific meetings: • Scott, M. P. (2020) The Role of Pectin Methyesterases in Gametophytic Incompatibility Systems in Maize [Abstract]. ASA, CSSA and SSSA International Annual Meetings (2020), Virtual (https://scisoc.confex.com/scisoc/2020am/meetingapp.cgi/Paper/126705) • Bapat, Amruta R, Moran Lauter, A.N. Hufford, M.B., Scott, M.P. 2020. Insights into the genetic architecture of the ga1 locus in maize genotypes Maize Genetics Conference Abstracts 62:P122, virtual. • URL: https://www.maizegdb.org/data_center/reference?id=3230341 Delate: • Because in-person field days were still restricted during the pandemic, a Virtual Field Day of an organic no-till corn experiment was developed as a YouTube video, which had 61 views, as of 10/15/21: https://www.youtube.com/watch?v=qyW0EfGG-Y&t=1s We continue to have buy-in from project farmers to grow the specific varieties or lines, and assist with data collection during the season. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Tracy: • Organology, Portland, OR. February 2019 • North Carolina State University, Raleigh, NC. June 2019 • Organic Seed Symposium, Corvallis, OR. August 2019 • Vitalis Organic Seed, San Juan Batista, CA. August 2019 • Organic Confluences. Baltimore MD. September 2019 • University of Minnesota, St. Paul, MN. September 2019. • Organic Seed Alliance, Port Townsend, WA. September 2019 • Association of University Technology Manager, St. Louis MO. September 2019. • Iowa Organic Conference Iowa City, IA, November 2019 • Diversifood, Rennes France. December 2019. • International Sweet Corn Development Association, Chicago, IL. December 2019. • Organic Seed Growers Conference, Corvallis, OR. February 2020. • Illinois Corn Breeders School, Champaign IL. March 2020. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1.Field corn, BS39-derived lines: From the tropical BS39 population that was adapted by Dr. Hallauer (ISU) to Iowa conditions, more than 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD). Testcrosses of the best 50 lines were evaluated under organic conditions in two locations (Ames, Iowa and Arlington, Wisconsin) in 2019, and in 2020 (Madison). Despite COVID-19 and a derecho that impacted Iowa, both 2020 trials were successfully completed, withdata analysis pending. In addition, a 2nd cycle breeding approach was adopted: the best two lines in terms of agronomic performance carrying the spontaneous haploid genome doubling (SHGD) ability were crossed with the 10 best lines with 100% BS39 background in summer 2019. Winter 2019/20, the haploids were induced for these crosses, and spontaneous doubling haploids selected summer 2020. More than 75 2nd generation inbreds with SHGD ability and 75% BS39 genome background were obtained. During summer 21 season, 38 of the 2nd cycle BS39-DH lines were grown in Ames in two row plots for a first evaluation. The lines show high variability, with plant heights between 0.9 and 1.79m, and flowering times varying between 1273 to 1556 GDD. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Development of donor lines in field and sweet corn carrying Ga1 and the major QTL for SHGD by marker-assisted backcrossing is proceeding as planned, and will be available by the end of this project. In Iowa and Wisconsin, sweet corn lines carrying the pollen exclusion locus Ga1 are being developed. These lines have been shown to exclude foreign pollen, making them well suited to organic production systems where foreign pollen can carry GMOs that eliminate the value added by organic production. In Iowa the derecho provided an opportunity for strong selection for stalk lodging. We learned that some of the germplasm developed for this program has moderate resistance to the emerging pest Fall Army Worm, providing plant breeders with a new resource to combat this pest. In addition, we proposed a molecular model to explain the function of the pollen exclusion systems we are using in this research. This model will help breeders develop effective deployment strategies for pollen exclusion systems. The development of 50% BS39 derived materials with SHGD and Ga1 is completed in seven different backgrounds. During S2021, haploids derived from the seven completed backgrounds were evaluated for their ability to spontaneously restore male fertility. The percentage of pollen shedding tassels in these directly seeded haploid populations varied between 30 and 81%. A similar experiment with BC3 derived sweet corn introgression lines showed, that MAS alone does not guarantee success for introducing the spontaneous doubling ability in these backgrounds. Phenotypic screening during the process is needed. Objectives 2, 3:Field plot design and data collection plans for the 2022 on-farm corn variety trials were finalized in-person with farmer-cooperators in March 2022. Results from the 2021 field corn season on four farms in Iowa (NE, NC, SW, NW) were impacted by wet weather in the spring, and drought in July and August. Despite the harsh weather, high yields were achieved across all farms, with no statistical yield differences between varieties. The ranking of yields was as follows: Blue River (Ames, IA) 64K93 at 195 bu/acre; Viking 0.74-10 (Albert Lea Seed, Albert Lea, MN) at 193 bu/acre; Prairie Hybrids (Deer Grove, IL) 5141 and 4211 at 192 bu/acre; and Viking O.51-04 and Blue River 54C27 at 190 bu/acre. At the research farm site in Greenfield, IA, the organic corn yields in the longest crop rotation sequence (C-S-O/A-A) averaged 177 bu/acre, compared to the conventional yields of 130 bu/acre. Ear samples were hand-collected from each variety at physiological maturity and measured for various parameters in 2021. Corn ear weight averaged 237 g/ear with no statistical differences among varieties, but the PH 5141 tended to be heavier, at 249 g/ear. Kernel weight per ear averaged 208 g/ear across all sites and varieties, with the PH 5141 tending to be heavier, at 217 g/ear. Ear length ranged from 19 to 22 cm across all varieties, with PH 5141 averaging the longest ears at 22 cm. Insect and disease damage ratings were very low on harvested ears in 2021, averaging 1 or lower (scale of 1 to 5, with 1 2% of ear damaged) across all varieties and sites. Objective 4. A graduate certificate in Organic Plant Breeding is offered at ISU.

Publications


    Progress 09/01/17 to 08/31/22

    Outputs
    Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?UW students worked with the Ga1 and Ga2 material with mentor and USDA collaborator Paul Scott in 2018, 2019, and 2020. ISU:Lauren Bilek and Megan Moore (undergraduate students), Yu-Ru Chen (Ph.D. student),and Josiah Pollock (Extension Program Specialist) were trained in on-farm experimental protocols and data analysis for this project. How have the results been disseminated to communities of interest?UW: Summer field days in Madison, WI 2019 and 2021 for organic growers. Information was presented at Midwest Food Processors Conference in Wisconsin Dells in 2019 and Organic Seed Growers Conference in Corvallis, OR in 2020. Other events interrupted by covid. ISU: besides project-specific activities, co-PI Delate is the organizer of the annual Iowa Organic Conference (November of each year in Iowa City), which allowed interactions of all project (co-)PIs and studentswith the organic community in Iowa and beyond. Field Days and conference presentations were utilized to disseminate information about the project. We continue to have buy-in from project farmers to grow the specific varieties or lines, and assist with data collection during the season. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. 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: In Iowa and Wisconsin, field and sweet corn lines carrying a pollen exclusion mechanism were developed, making them well-suited fororganic production systems. These linesexcludeforeign pollen that can carry GMOs, which wouldeliminate the value of organically produced seed. Moreover, agenetic mechanism for accelerated inbred line development without the need to use chemicals, as done in comparable conventional procedures, was successfully established. Both field and sweet corn prototypes were developed in this project, which can be used as donors of these genetic mechanisms in public and private organic maize breeding programs. This materialis being used in twoUSDA OREI follow-up projects led by Drs. Scott (COOP) and Lubberstedt (CoMGI). Results from this project (ACES) were disseminated by various publications, conferences, field days, and the National Association of Plant Breeders meeting held in Ames, IA, 2022, co-hosted by (co-)PIs Lubberstedt and Scott. A graduate certificate program in Organic Plant Breeding was established and is offered at ISU. Objective1. Field corn, BS39-derived lines: From the tropical BS39 population that was adapted by Dr. Hallauer (ISU) to Iowa conditions, more than 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD). Testcrosses of the best 50 lines were evaluated under organic conditions in two locations (Ames, Iowa and Arlington, Wisconsin) in 2019and 2020 (Madison). Despite COVID-19 and a derecho that impacted Iowa, both 2020 trials were successfully completed, data analysis pending. In addition, a 2nd cycle breeding approach was adopted: the best two lines in terms of agronomic performance carrying the spontaneous haploid genome doubling (SHGD) ability were crossed with the 10 best lines with 100% BS39 background in summer 2019. Winter 2019/20, the haploids were induced for these crosses, and spontaneous doubling haploids selected summer 2020. More than 75 2nd generation inbreds with SHGD ability and 75% BS39 genome background were obtained. During summer 21 season, 38 of the 2nd cycle BS39-DH lines were grown in Ames in two row plots for a first evaluation. The lines show high variability, with plant heights between 0.9 and 1.79m, and flowering times varying between 1273 to 1556 GDD. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Development of donor lines in field and sweet corn carrying Ga1 and the major QTL for SHGD by marker-assisted backcrossing is proceeding as planned, and will be available by the end of this project. In Iowa and Wisconsin, sweet corn lines carrying the pollen exclusion locus Ga1 are being developed. These lines have been shown to exclude foreign pollen, making them well-suited to organic production systems where foreign pollen can carry GMOs that eliminate the value added by organic production. In Iowa, the derecho provided an opportunity for strong selection for stalk lodging. We learned that some of the germplasm developed for this program has moderate resistance to the emerging pest Fall Army Worm, providing plant breeders with a new resource to combat this pest. In addition, we proposed a molecular model to explain the function of the pollen exclusion systems we are using in this research. This model will help breeders develop effective deployment strategies for pollen exclusion systems. The development of 50% BS39 derived materials with SHGD and Ga1 is completed in seven different backgrounds. During S2021, haploids derived from the seven completed backgrounds were evaluated for their ability to spontaneously restore male fertility. The percentage of pollen shedding tassels in these directly seeded haploid populations varied between 30 and 81%. A similar experiment with BC3 derived sweet corn introgression lines showed that MAS alone does not guarantee success for introducing the spontaneous doubling ability in these backgrounds. Phenotypic screening during the process is needed. Objectives 2 and3: Field plot design and data collection plans for the 2022 on-farm corn variety trials were finalized in-person with farmer-cooperators in March 2022. Results from the 2021 field corn season on four farms in Iowa (NE, NC, SW, NW) were impacted by wet weather in the spring, and drought in July and August. Despite the harsh weather, high yields were achieved across all farms, with no statistical yield differences between varieties. The ranking of yields was as follows: Blue River (Ames, IA) 64K93 at 195 bu/acre; Viking 0.74-10 (Albert Lea Seed, Albert Lea, MN) at 193 bu/acre; Prairie Hybrids (Deer Grove, IL) 5141 and 4211 at 192 bu/acre; and Viking O.51-04 and Blue River 54C27 at 190 bu/acre. At the research farm site in Greenfield, IA, the organic corn yields in the longest crop rotation sequence (C-S-O/A-A) averaged 177 bu/acre, compared to the conventional yields of 130 bu/acre. Ear samples were hand-collected from each variety at physiological maturity and measured for various parameters in 2021. Corn ear weight averaged 237 g/ear with no statistical differences among varieties, but the PH 5141 tended to be heavier, at 249 g/ear. Kernel weight per ear averaged 208 g/ear across all sites and varieties, with the PH 5141 tending to be heavier, at 217 g/ear. Ear length ranged from 19 to 22 cm across all varieties, with PH 5141 averaging the longest ears at 22 cm. Insect and disease damage ratings were very low on harvested ears in 2021, averaging 1 or lower (scale of 1 to 5, with 1 2% of ear damaged) across all varieties and sites. Objective 4. A graduate certificate program in Organic Plant Breeding is offered at ISU.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2021 Citation: McCluskey, C. & Tracy, W.F. (2021). Engaging Farmer Stakeholders: Maize Producers Perceptions and Strategies for Managing On-Farm Genetic Diversity in the Upper Midwest. Sustainability, 13: 8843, https://www.mdpi.com/2071-1050/13/16/8843/htm
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Revilla, P., Anibas, C.M. & Tracy, W.F. (2021). Sweet Corn research around the world 2015 2020. Agronomy 11(34). https://doi.org/10.3390/agronomy11030534


    Progress 09/01/20 to 08/31/21

    Outputs
    Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems:Because of the derecho in 2020, we will use another year for on-farm trials, which will primarily require efforts from co-PI Delate. Cost-neutral extension was granted, which will in addition allow completion of data analysis and line conversion efforts by the other co-PIs. What opportunities for training and professional development has the project provided? Lubberstedt: New PhD students Yu-Ru Chen and Mercy Fakude (Fulbright fellow) joined this project. Yu-Ru Chen gave poster presentations at the RF Baker symposium in Ames, IA, 2021, and at the virtual 2021 NAPB meeting. Scott: Trained Iowa State University Ph.D. Students Amruta Bapat and Nicholas Boerman. Trained undergraduate laboratory assistant Hannah Clubb. Tracy: MS student Cathleen McClusky finished her MS program, and continues as PhD student with Dr. Tracy. Delate:Henry Franzen, undergraduate student, was trained in on-farm experimental protocols and data analysis for this project. How have the results been disseminated to communities of interest? Lubberstedt (lecture): Lubberstedt, T. Past, present and future of maize doubled haploid technology. Zemun Polje conference, Belgrade Serbia (virtual) June 8, 2021. Poster presentations of group members (see above). Scott: Information was disseminated in the following Lectures: Scott, M.P. Using Gametophytic Incompatibility Systems to Improve Genetic Purity of Specialty Maize. Zemun Polje conference, Belgrade Serbia (virtual) June 8, 2021. Scott, M.P. Breeding corn for organic production systems. Iowa State University Plant Breeding Seminar, May 5, 2021. Scott, M.P. Cross incompatibility in Corn. Iowa State University Plant Biology Faculty Seminar. September 29, 2021. Information was disseminated through the following posters at virtual scientific meetings: Scott, M. P.(2020) The Role of Pectin Methyesterases in Gametophytic Incompatibility Systems in Maize [Abstract]. ASA, CSSA and SSSA International Annual Meetings (2020), Virtual (https://scisoc.confex.com/scisoc/2020am/meetingapp.cgi/Paper/126705) Bapat, Amruta R, Moran Lauter, A.N. Hufford, M.B., Scott, M.P. 2020. Insights into the genetic architecture of the ga1 locus in maize genotypes Maize Genetics Conference Abstracts 62:P122, virtual. URL: https://www.maizegdb.org/data_center/reference?id=3230341 Delate: Because in-person field days were still restricted during the pandemic, a Virtual Field Day of an organic no-till corn experiment was developed as a YouTube video, which had 61 views, as of 10/15/21: https://www.youtube.com/watch?v=q-yW0EfGG-Y&t=1s We continue to have buy-in from project farmers to grow the specific varieties or lines, and assist with data collection during the season. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Tracy: Organology, Portland, OR. February 2019 North Carolina State University, Raleigh, NC. June 2019 Organic Seed Symposium, Corvallis, OR. August 2019 Vitalis Organic Seed, San Juan Batista, CA. August 2019 Organic Confluences. Baltimore MD. September 2019 University of Minnesota, St. Paul, MN. September 2019. Organic Seed Alliance, Port Townsend, WA. September 2019 Association of University Technology Manager, St. Louis MO. September 2019. Iowa Organic Conference Iowa City, IA, November 2019 Diversifood, Rennes France. December 2019. International Sweet Corn Development Association, Chicago, IL. December 2019. Organic Seed Growers Conference, Corvallis, OR. February 2020. Illinois Corn Breeders School, Champaign IL. March 2020. What do you plan to do during the next reporting period to accomplish the goals?Lubberstedt and Scott: Will co-host the 2022 National Association of Plant Breeders meeting at Iowa State University (August 8-11, 2022). This in-person event will offer the possibility to engage with stakeholders, andto show-case outcomes of this USDA OREI project, e.g., during the planned field tour, and by student poster presentations. Lubberstedt: Will finish analysis of data from this project for publication. Moreover, development of field and sweet corn lines carrying SHGD and Ga1 will be completed. This materials will be used in ongoing USDA OREI projects COOP and CoMGI. Delate: Hybrids from germplasm developed under the 2014 NIFA - OREI grant, and any germplasm that becomes available from this OREI grant, will be tested in organic farmer-cooperators' fields. Sweetcorn germplasm from UW's program will also be tested. Field Days will be held on two on-farm sites in 2022.

    Impacts
    What was accomplished under these goals? Objective 1.Develop novel organic field corn and sweet corn genotypes for the commercialization pipeline that utilize more efficient methods of breeding. Field corn, BS39-derived lines: From the tropical BS39 population that was adapted by Dr. Hallauer (ISU) to Iowa conditions, more than 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD) (Verzegnazzi et al. 2021; Santos et al. 2021). A 2nd cycle breeding approach was adopted: the best two lines in terms of agronomic performance carrying SHGDability were crossed with the 10 best lines with 100% BS39 background. Winter 2019/20, the haploids were induced for these crosses, and spontaneous doubling haploids selected summer 2020. More than 75 2nd generation inbreds with SHGD ability and 75% BS39 genome background were obtained, seed was multiplied winter 2020/21. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Development of donor lines in field and sweet corn carrying Ga1 and the major QTL for SHGD by marker-assisted backcrossing is proceeding as planned, and will be available by end of this project. In Iowa and Wisconsin, sweet corn lines carrying the pollen exclusion locus Ga1 are being developed. These lines have been shown to exclude foreign pollen, making them well suited to organic production systems where foreign pollen can carry GMOs that eliminate the value added by organic production. In Iowa the derecho provided an opportunity for strong selection for stalk lodging. We learned that some of the germplasm developed for this program has moderate resistance to the emerging pest Fall Army Worm, providing plant breeders with a new resource to combat this pest. In addition, we proposed a molecular model to explain the function of the pollen exclusion systems we are using in this research. This model will help breeders develop effective deployment strategies for pollen exclusion systems. 2021 progress Field corn, BS39-derived lines:During summer 21 season, 38 of the 2nd cycle BS39-DH lines were grown in Ames (and by collaborator Dr. Bohn, UIUC in Champaign) in 2-row plots. Lines were uniform, plant heights varyingbetween 0.9 and 1.79m, and flowering times between 1273 to 1556 GDD. Data from UIUC have been provided and will be jointly analyzed. In the frame of the USDA OREI project CoMGI, crosses conducted between those lines will be sent to the winter nursery in Puerto Rico, and haploids induced. Haploids will be planted in Ames '22,to develop improved BS39-derived breeding materials. Introduction of SHGD and Ga1 into field and sweet corn (ISU): The development of 50% BS39 derived materials with SHGD and Ga1 is completed in seven different backgrounds, and still ongoing for another three backgrounds. During S2021, haploids derived from the seven completed backgrounds were evaluated for their ability to spontaneously restore male fertility. The percentage of pollen shedding tassels in these directly seeded haploid populations varied between 30 and 81%. A similar experiment with BC3 derived sweet corn introgression lines showed, that MAS alone does not guarantee success for introducing the spontaneous doubling ability in these backgrounds. Phenotypic screening during the process is needed. The SHGD donor was crossed to seven sweet corn inbred lines, and the respective F1s will be backcrossed to the respective inbred lines (Dr. Tracy) and haploid induced (ISU DH Facility) during winter of 2021/22. Resulting haploids will be evaluated for SHGD during summer of 2022. We are developing sweetcorn with improved genetic purity by transferring the Ga2 allele into sweet corn lines. This year we advanced breeding lines with selection for the ability to exclude unwanted pollen and agronomic properties. Interestingly, we observed several lines that were unexpectedly able to cross with lines that should have excluded their pollen, suggesting that our model for function of gametophytic incompatibility systems requires revision. We developed a new hypothesis to explain these data. We suspect that there is interaction between different gametophytic incompatibility systems. An alternative hypothesis is that the function of gametophytic incompatibility systems is dependent on environmental conditions. We have initiated experiments to test these hypotheses. These results will enable to develop corn varieties with improved genetic purity, reducing the impact of GMO contamination on organic corn producers. Objective 2.Select and test on-farm genotypes that meet the critical needs of organic producers. In 2021, six commercial organic field corn varieties (BR 54C27, BR 64K93, Prairie Hybrids 4211, Prairie Hybrids 5141, Viking 0.51-04, and Viking 0.74-10) were compared, with one farm (NC) also evaluating the USDA hybrid, 5323219. Plant emergence across the four on-farm sites ranged from 28,300 plants/acre at the NE site to 33,667 plants/acre at the SW site. Mid-season commercial organic corn plant height varied across varieties and sites, with the Blue River 54C27 variety the tallest at 273 cm, compared to the lowest average height in the Prairie Hybrids 5141 variety at 257 cm. Weed populations were managed well in 2021 across all sites with grass weeds in mid-June ranging from 0 weeds/sq. foot at the SW site to 6 weeds/sq. foot at the NE site. Broadleaf weeds averaged 0 weed/sq. foot at the SW site to 2 weeds/sq. foot at the NE site. Correlations between weed populations and yields will be conducted after all plots are harvested in 2021. Objective 3.Practice technology transfer through a network with organic farmers and organic seed companies. Field plot design and data collection plans for the 2021 on-farm corn variety trials were finalized in-person with farmer-cooperators. The 2020 field corn season on four farms in Iowa (NE, NC, SW, NW) was very challenging, with wet weather in the spring, drought in July and August, and a derecho in August. Despite the harsh weather, Viking 0.18-06 UP (Albert Lea Seed, Albert Lea, MN) and Viking 0.51-04 produced the highest yields, at 168 bu/acre and 166 bu/acre, respectively, across the four on-farm sites. Blue River 48G35 and 57A30 varieties averaged 161 bu/ac and 163 bu/ac, respectively. A USDA hybrid (19MPS/19SJWF) showed competitiveness with commercial hybrids, at 147 bu/acre; the other USDA hybrid (19SJWE/185MPS) was lower at 114 bu/acre. Despite 100 mph winds from the derecho, commercial organic hybrid yields at the most affected farm (North Central Iowa) were exceptional, averaging 165 bu/acre. Ear samples were hand-collected from each variety at physiological maturity and measured for various parameters in 2020. Corn ear weight averaged 200 g/ear for the BR57A30 variety, while the Viking 0.18-06 UP averaged 239 g/ear across all sites. Kernel weight per ear averaged 172 g/ear for the BR48G35 variety while the Viking 0.18-06 UP averaged 209 g/ear across all sites. Ear length ranged from 18 to 21 cm, with Viking 0.18-06 UP averaging the longest ears at 21 cm. Kernel weight in the USDA varieties ranged from 175 to 192 g/ear, with ear length ranging from 18 to 19 cm. Insect and disease damage ratings were very low on harvested ears in 2020, averaging less than 1 (scale of 1 to 5, with 1 2% of ear damaged) across all varieties and sites. Objective 4.Develop a graduate Certificate program in Organic Plant Breeding. The "Organic Plant Breeding" certificate program is available at ISU (https://www.agron.iastate.edu/graduate-department/new-organic-plant-breeding-certificate). A student in the Distance MS program at ISU (Christy Montes) developed materials in her creative component (defense 11/2020), which will be used in the organic plant breeding course. Another student (Lawrence Warnock)will do the same in prepare his creative component spring 2022.

    Publications

    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Delate, K. (2021) Surviving COVID, Derecho, and Drought Lessons from2020- Midwest Organic and Sustainable Education Service (MOSES) Organic Conference, La Crosse, WI, 2/25/21 https://www.dropbox.com/s/lgceptpiwih34vp/Surviving%20COVID%2C%20Derecho%2C%20and%20Drought%20Lessons%20of%20Resilience%20in%202020%20edited.mp4?dl=0
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Delate, K. (2021) Risk Management for Organic Production: March 10, 2021, ISU-FFED, Ames, IA https://www.extension.iastate.edu/ffed/specialty-crops-risk-management-webinar-series/
    • Type: Other Status: Other Year Published: 2021 Citation: Delate, K. (2021) Grain Place Foundation Field Day, July 17, 2021 http://www.grainplacefoundation.org/news/
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Delate, K. (2021) NRCS Webinar: Organic Research to Support Transitioning, February 18, 2021, Des Moines, IA
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Verzegnazzi, A., Goncalves, I., Frei, U.K., Krause, M., Campbell, J., Almeida, V., Tonello Zuffo, L., Boerman, N., L�bberstedt, T. (2021) Major locus for spontaneous haploid genome doubling detected by a case-control GWAS enables efficient doubled haploid line development in exotic maize germplasm. Theor. Appl. Genet. 134: 1423-1434 DOI: 10.1007/s00122-021-03780-8
    • Type: Journal Articles Status: Other Year Published: 2021 Citation: Goncalves, I., Verzegnazzi, A.L., Edwards, J., Frei, U.K., De La Fuente, G.N., Zuffo, L., Pires, L.P.M., L�bberstedt, T. (2021) Usefulness of Adapted Exotic Maize Lines Developed By Doubled Haploid and Single Seed Descent Methods. (https://www.researchsquare.com/article/rs-799789/v1)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Thomas L�bberstedt, Siddique Muhammad-Aboobucker, Tanner Cook, Yu-Ru Chen, Mercy Fakude, Liming Zhou, Elizabeth Bovenmyer, Sarah Pfeffer, Ursula Karoline Frei. Past, present, and future of maize doubled haploid technology. Conference "The Frontiers of Science and Technology in Crop Breeding and Production", Zemun Polje, Belgrade.
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zystro, Jared, Tessa Peters, Kathleen Miller, and William F. Tracy 2021. Inbred and hybrid sweetcorn genotype performance in diverse organic environments. Crop Science 10.1002/csc2.20457
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Solemslie, R., du Toit, L.J., Tracy, W.F., and Stearns, T. 2021. Evaluation of steam treatments for Fusarium spp. and other fungi on sweet corn seed, 2021. Plant Disease Management Reports 15:CF017.
    • Type: Journal Articles Status: Accepted Year Published: 2021 Citation: Muhammad-Aboobucker, S., Jubery, Z., Frei, U.K., Foster, T., Chen, Y.-R., Ganapathysubramanian, B., L�bberstedt, T. (2021) Protocols for in vivo doubled haploid (DH) technology in maize breeding: From haploid inducer to haploid genome doubling. Methods Molecular Biology (in press)
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zystro, Jared, Tessa Peters, Kathleen Miller, and William F. Tracy 2021. Classical and genomic prediction of synthetic open pollinated sweet corn performance in organic environments. Crop Science https://doi.org/10.1002/csc2.20531
    • Type: Journal Articles Status: Published Year Published: 2021 Citation: McCluskey, C. and W.F. Tracy. 2021. Engaging Farmer Stakeholders: Maize Producers Perceptions and Strategies for Managing On-Farm Genetic Diversity in the Upper Midwest. Sustainability https://www.mdpi.com/2071-1050/13/16/8843/htm
    • Type: Journal Articles Status: Accepted Year Published: 2021 Citation: Boerman, N., Moran Lauter, A.N., Scott, M.P. (2020) Variation in degree of pollen exclusion for Ga1-s unilateral cross incompatibility in temperate maize breeding populations, Agrosystems, Geosciences & Environment (In Press).
    • Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Boerman, N, (2020) Genetic analysis of doubled haploid and unilateral cross incompatibility systems for organic maize breeding, Ph.D. Thesis Iowa State University Department of Agronomy.
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Zystro, Jared, Tessa Peters, Kathleen Miller, and William F. Tracy 2020. Classical and genomic prediction of hybrid sweet corn performance in organic environments. Crop Science 10.1002/csc2.20400
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Moore, Virginia M. and Tracy, William F. 2020. Combining ability of husk extension, maysin content, and corn earworm resistance. Journal of American Society of Horticultural Science 146:14-23.
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Moore, Virginia M. and Tracy, William F. 2020. Survey of organic sweet corn growers identifies corn earworm prevalence, management, and opportunities for plant breeding. Renewable Agriculture and Food Systems pp. 1  4 DOI: https://doi.org/10.1017/S1742170520000204
    • Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: McCluskey, C.A. 2020. Corn in the upper Midwest: farmers perceptions and strategies for managing on-farm genetic diversity. Master of Science Thesis. University of Wisconsin-Madison
    • Type: Other Status: Published Year Published: 2020 Citation: Delate, K. (2020) Small Farm Sustainability Podcast: Iowa Organic Update, November 11, 2020, Iowa State University, Ames, IA https://www.extension.iastate.edu/smallfarms/small-farm-sustainability-podcasts
    • Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Delate, K. (2020) Hort 530 Lecture: Update on Organic Research: Nov. 17, 2020, ISU Dept. of Horticulture, Ames, IA
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. (2020) Iowa Organic Conference, Nov. 23, 2020, ISU Dept. of Horticulture, Ames, IA: https://www.iowaorganic.org/2020_iowa_organic_conference
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. (2020) OATS (Organic Agronomy Training Series) Podcast: The Dirt on Organic Farming https://www.organicagronomy.org/the-dirt-on-organic-farming


    Progress 09/01/19 to 08/31/20

    Outputs
    Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems:2020 certainly was/is an unusual year severely impacted by the COVID-19 pandemia, and in Iowa: a major wind event (derecho). This led to university closures after spring break 2020, and reduced research activities in laboratories and research farms. Nonetheless, both ISU, USDA and U Wisconsin were able to continue their field research and nursery activities, while overall reduced, field activities for this project were not affected and continued as planned. The derecho reduced nursery harvest at ISU somewhat, but not dramatically. The BS39 field trial at ISU was impacted, but again, the wind event may even be helpful to differentiate lines with regard to lodging tolerance. What opportunities for training and professional development has the project provided? Overall, opportunities in 2020 were more limited due to the COVID-19 pandemic. One of the last in person events was the RF Baker Symposium at ISU with student poster presentations. Amruta Bapat is an Iowa State University Graduate student in the Interdepartmental Genetics and Genomics degree program. She carried out research on the molecular genetics of gametophytic incompatibility. She presented her results in a poster at the 2020 virtual Maize Genetics Conference. Nicholas (Nick) Boerman is an Iowa State University graduate student in the plant breeding program, and gave an oral presentation at the ASA/CSSA meeting 2019 in San Antonio. Dr. Tracy trained Ph.D. student Cathleen McCluskey. Cathleen McCluskey is a masters degree candidate in theinterdepartmental Agroecology Program at University of Wisconsin-Madison. In this project, she receives training in maize genetics, disease identification, experimental design, statistics, and nursery operation, including planting, pollinating, and harvesting seed. Her thesis work involves measuring and assessing genetic diversity in U.S. maize, which resulted in a MS thesis in 2020. Drs. Henrique Trentin and Arthur Pereira were graduate students in the plant breeding program at ISU. Even though not covered by this project, both contributed to research relevant for this project. Henrique was involved in haploid inducer development, and Arthur in studying spontaneous haploid genome doubling. Both defended in 2019, and are meanwhile working in their home country Brazil for Bayer and Stine, respectively. How have the results been disseminated to communities of interest?Because no in-person field days were permitted during the pandemic, a Virtual Field Day of the organic experiments at the ISU Neely-Kinyon Farm in Greenfield, Iowa, was developed as a YouTube video, which had 298 views, as of 10/15/20: https://www.youtube.com/watch?v=P_PplV5EmT4 We continue to have buy-in from project farmers to grow the specific varieties or lines, and assist with data collection during the season. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Results were published in scientific journals. Other presentations: Scott Presented invited talk at University of Illinois, Urbana -Champaign "Breeding corn for organic production systems" November 21, 2019. Poster at organic seed growers conference Poster at MOSES What do you plan to do during the next reporting period to accomplish the goals? Lubberstedt: Analyze BS39 data, evaluate 2nd cycle lines in summer of 2021. Initiate the certificate in organic plant breeding. Scott: We plan to advance Ga2 breeding lines with selection for Agronomic traits and pollen exclusion. We will continue to characterize pollen exclusion systems both biochemically and genetically to gain a better understanding of how to use these systems to preserve the genetic purity of corn varieties. Delate: Hybrids from germplasm developed under the 2014 NIFA - OREI grant, and any germplasm that becomes available from this OREI grant, will be tested in organic farmer-cooperators' fields. Sweetcorn germplasm from UW's program will also be tested. Field Days will be held on at least two on-farm sites in 2021. Tracy: Wisconsin will advance Ga1 material and continue to evaluate efficacy of pollen exclusion in sweet corn. We will continue working with Ames on moving the SHGD into sweet corn.

    Impacts
    What was accomplished under these goals? Overall impact statement: The overall goal of the project is to substantially increase the productivity of organic field and sweet corn production within organic crop rotations, by improving the efficiency of breeding through doubled haploid (DH) technology and traditional breeding methods, focused on early plant vigor and competitiveness factors. Another goal is to develop a Certificate program in Organic Plant Breeding, which can be accessed online in the U.S. and internationally. All project partners conducted field experiments for evaluation of maize lines under organic conditions, and / or breeding work to develop novel germplasm. For example, 400 lines developed for this project were evaluated in six environments. Superior lines with regard to agronomic performance under organic conditions and for quality traits were identified. These superior lines will receive by conventional crosses additional properties which will make future breeding of organic maize varieties more efficient. A major finding was that an important property for DH technology is simple inherited (one single major gene), which will simplify breeding strategies to incorporate this property into elite breeding materials. Our findings will lead to changes in knowledge and action in the way breeding of organic field and sweet corn will be done in future, by using DH technology and the novel genetic property characterized in this project. Outcomes will be first generation field and sweet corn lines carrying this property along with another important trait (cross-incompatibility) employed in this project. Stakeholders will be engaged to evaluate first prototypes under organic practise conditions, to determine robustness of this materials. The certificate program on Organic Plant Breeding to be established in this program will be a venue, to share new knowledge in the field or organic plant breeding with students, working professionals, and others engaged in this topic. Strengthened organic plant breeding will lead to better adapted and more competitive varieties under organic conditions, which benefits farmers but also the general public, as this should impact prices of organic products. Objective 1:Develop novel organic field corn and sweet corn genotypes for the commercialization pipeline that utilize more efficient methods of breeding. Field corn, BS39-derived lines: From the tropical BS39 population that was adapted by Dr. Hallauer (ISU) to Iowa conditions, more than 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD). Testcrosses of the best 50 lines were evaluated under organic conditions in two locations (Ames, Iowa and Arlington, Wisconsin) in 2019, and in 2020 (Madison). Despite of COVID-19 and a derecho that impacted Iowa, both 2020 trials were successfully completed, data analysis pending. In addition, a 2nd cycle breeding approach was adopted: the best two lines in terms of agronomic performance carrying the spontaneous haploid genome doubling (SHGD) ability were crossed with the 10 best lines with 100% BS39 background in summer 2019. Winter 2019/20, the haploids were induced for these crosses, and spontaneous doubling haploids selected summer 2020. More than 75 2nd generation inbreds with SHGD ability and 75% BS39 genome background were obtained, seed will be multiplied winter 2020/21, and respective inbreds be evaluated within this project during summer of 2021. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Development of donor lines in field and sweet corn carrying Ga1 and the major QTL for SHGD by marker-assisted backcrossing is proceeding as planned, and will be available by end of this project. In Iowa and Wisconsin, sweet corn lines carrying the pollen exclusion locus Ga2 are being developed.These lines have been shown to exclude foreign pollen, making them well suited to organic production systems where foreign pollen can carry GMOs that eliminate the value added by organic production. In Iowa the derecho provided an opportunity for strong selection for stalk lodging. We learned that some of the germplasm developed for this program has moderate resistance to the emerging pest Fall Army Worm, providing plant breeders with a new resource to combat this pest. In addition, we proposed a molecular model to explain the function of the pollen exclusion systems we are using in this research. This model will help breeders develop effective deployment strategies for pollen exclusion systems. Objective 2:Select and test on-farm genotypes that meet the critical needs of organic producers. Seed production efforts were limited by the pandemic this year. We produced seed of one hybrid for distribution to farmers. Objective 3:Practice technology transfer through a network with organic farmers and organic seed companies. Field plot design and data collection plans for the 2020 on-farm corn variety trials finalized at a project farmer meeting at the MOSES conference on February 29, 2020. The 2020 season on four farms in Iowa (NE, NC, SW, NW) was very challenging, with wet weather in the spring, drought in July and August, and an inland hurricane (derecho) with 100 mph winds on August 10, that seriously affected the NC on-farm sites in Jefferson, Iowa. Plant emergence was excellent in 2020, with no statistical differences in plant populations among the commercial organic field corn varieties (BR 57A30, BR 48G35, Viking 0.18-06 UP and Viking 51-04), ranging from 30,438 plants/acre at the NE site to 38,250 plants/acre at the SW site. Late-July commercial organic variety plant height varied across varieties and sites, with the Viking 51-04 averaging 248 cm across three sites, compared to the 190-cm height in the BR48G35 at the NC site. Weed populations were well managed in 2020 across all sites. Grass weeds mid-June ranged from 2 weeds/sq. meter at the SW site, to 21 weeds/sq. meter at the NE site. Broadleaf weeds averaged 1 weed/sq. meter at the NE site to 5 weeds/sq. meter at the SW site. Correlations between weed populations and yields will be conducted after all plots are harvested. The USDA-bred corn (19SMPS/19SJWF and 19SFWE/185MPS) was only grown at the NC site, where there were with no statistical differences in plant populations, averaging 33,500 plants/acre. Late-July 19SMPS/19SJWF plant height averaged 157 cm while 19SFWE/185MPS averaged 182 cm. Weed populations were very low at this site, with mid-June grass weeds averaging 4 weeds/sq. meter and broadleaf weeds averaging 1 weed/sq. meter. Corn stalk nitrate samples collected from on-farm sites were analyzed by a commercial lab. Stalk nitrate content ranged from 218 ppm in the Viking 18-06 variety to 1,667 ppm in the BR57A30 variety. Ear samples were hand-collected from each variety at physiological maturity and measured for various parameters. Corn ear weight ranged from 101 g/ear for the BR48G35 variety at the NW site to 273 g/ear for Viking 18-06 at the NE site. Kernel weight per ear ranged from 88.2 g/ear for the BR48G35 variety at the NW site to 238 g for Viking 18-06 at the NE site. Kernel weight in the USDA varieties ranged from 175 to 192 g/ear, with ear length ranging from 18 to 19 cm. As in previous years, with the BR 57A30 producing the greatest yield in 2019, at 233 bu/acre, at the NW site. Objective 4: Develop a graduate Certificate program in Organic Plant Breeding. The Certificate program "Breeding for Organic Crops" havebeen approved byISU and will be offered from 2021 on, as three courses are already available, while the "Organic Plant Breeding" course (as last course in the program) willbe developed in 2021. A student in the Distance MS program at ISU (Christy Montes) developed materials in her creative component (defense 11/2020), which will be used in the organic plant breeding course.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Coffman, S.M., Hufford, M., Andorf, C., L�bberstedt, T. (2020) Diversity of haplotype structure among commercial maize ex-PVP lines in relation to key pre-commercial germplasm. Theor. Appl. Genet. 133:547-561 https://doi.org/10.1007/s00122-019-03486-y
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ren, J., Boerman, N., Liu, R., Frei, U.K., Trampe, B., Vanous, K., Chen, S., L�bberstedt, T. (2020) QTL mapping of spontaneous haploid genome doubling. Plant Sci. 293:110337 https://doi.org/10.1016/j.plantsci.2019.110337
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Trampe, B., Goncalves, I., Frei, U.K., Ren, J., Chen, S., L�bberstedt, T. (2020) QTL mapping of Spontaneous Haploid Genome Doubling using Genotype by Sequencing Approach in maize. Theor. Appl. Genet. 133:2131-2140 https://doi.org/10.1007/s00122-020-03585-1
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Gustin, J.L., Frei, U.K., Baier, J., Armstrong, P., L�bberstedt, T., Settles, A.M. (2020) Maize haploid classification using Single Kernel Near-Infrared Spectroscopy. Plant Breeding (in press) DOI: 10.1111/pbr.12857
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Abel, C. A., Coates, B. S., Millard, M., Williams, W. P. & Scott, M. P. Evaluation of XL370A-Derived Maize Germplasm for Resistance to Leaf Feeding by Fall Armyworm. Southwest. Entomol. 45, 6974 (2020).
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ma, L., Qing, C., Frei, U.K., Shen, Y., L�bberstedt, T. (2020) Association mapping for root system architecture traits under different nitrogen conditions in germplasm enhancement of maize doubled haploid (GEM-DH) lines. The Crop J 8:213-226 https://doi.org/10.1016/j.cj.2019.11.004
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Boerman, N.A., Frei, U.K., L�bberstedt, T. (2020) Impact of Spontaneous Haploid Genome Doubling in Maize Breeding. Plants 9: 369
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Trentin, H.U., Frei, U.K., L�bberstedt, T. (2020) Maternal haploid inducer development in maize. Plants 9: 614 doi:10.3390/plants9050614
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Qu, Y., Wu, P., Ren, J., Liu, Z., Tang, J., L�bberstedt, T., Chen, S., Li, H. (2020) Mapping of QTL for kernel abortion caused by in vivo haploid induction in maize (Zea mays L.). PloS One 15: e0228411
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: De la Fuente, G., Frei, U.K., Trampe, B., Ren, J., Bohn, M.O., Yana, N., Verzegnazzi, A., Murray, S.C., L�bberstedt, T. (2020) A diallel analysis of a maize donor population response to in vivo maternal haploid induction. II: Spontaneous Haploid Genome Doubling. Crop Sci. 60: 873-882 DOI: 10.1002/csc2.20021
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Costa Almeida, V., Trentin, H.U., Frei, U.K., L�bberstedt, T. (2020) Genomic prediction in maternal haploid induction in maize. The Plant Genome 13:e20014 DOI: 10.1002/tpg2.20014
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: Lu, Y., Moran Lauter, A. N., Makkena, S., Scott, M. P. & Evans, M. M. S. Insights into the molecular control of cross-incompatibility in Zea mays. Plant Reprod. (2020) doi:10.1007/s00497-020-00394-w.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Bapat, Amruta R, Moran Lauter, AN, Hufford, MB, Scott, MP. 2020. Insights into the genetic architecture of the ga1 locus in maize genotypes. Maize Genetics Conference Abstracts 62:P122. https://www.maizegdb.org/data_center/reference?id=3230341
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. (2020) On-Farm Organic Corn Trials  Farmer-Cooperator Meeting, Midwest Organic and Sustainable Education Service (MOSES) Organic Conference, La Crosse, WI, 2/29/20
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. (2020) Organic Research in Iowa - Horticulture Dept., University of Florida, Gainesville, FL, 3/18/20 (via Zoom)
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Delate, K., T. Lubberstedt, P. Scott, W. Tracy and R. Turnbull. 2019. Developing field and sweet corn hybrids for organic producers: The ACES project. ASA Annual Meeting, San Antonio, TX. https://scisoc.confex.com/scisoc/2019am/meetingapp.cgi/Paper/120078
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Delate, K. 2019. Developing field and sweet corn hybrids for organic producers: The ACES project. ASA Annual Meeting, San Antonio, TX. Presented Nov. 11, 2019.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. 2020. Video: Virtual Neely-Kinyon Farm Organic Field Day. Available at: https://www.youtube.com/watch?v=P_PplV5EmT4
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. 2020. Virtual Organic Field Day Available On-Line. ISU Extension and Outreach, September 11, 2020. Available at: https://www.extension.iastate.edu/news/virtual-organic-field-day-video-available-online
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Delate, K. 2020. Organic Agriculture Concerns Amid COVID-19. ISU Extension and Outreach, May 5, 2020. Available at: https://www.extension.iastate.edu/news/organic-agriculture-production-concerns-amid-covid-19
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: McCluskey, C. W.F.Tracy, Midwest Organic and Sustainable Education Services (MOSES) Organic Farming Conference Research Poster Session, La Crosse, WI, Midwest Corn Growers' Perceptions and Management Strategies of On-Farm Genetic Diversity.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: McCluskey, C. W.F. Tracy Selections (ACES) Organic Breeding Program: Novel Strategies to Develop Field & Sweet Corn for Organic Producers (Proceedings at https://seedalliance.org/publications/proceedings-from-the-10th-organic-seed-growers-conference/)
    • Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: McCluskey, C. Master of Science in Agroecology at the University of Wisconsin-Madison, Corn in the Upper Midwest Farmers' Perceptions and Strategies for Managing On-Farm Genetic Diversity


    Progress 09/01/18 to 08/31/19

    Outputs
    Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems:Objective 1: after testing 400 inbred lines derived from the BS39 population in 2018, we noticed wide genetic variation for agronomic traits in this tropical-adapted materials. We decided, after discussions among project participants and with board members December 2018, to deviate from the original approach to re-test all 400 lines in 2019. Instead, we employed a breeder's approach of re-testing only the 50 best lines, while already producing the 2nd cycle of breeding materials after crossing the best lines. 2nd cycle DH lines will be available in 2020, and evaluated in the last year of this project in 2021. What opportunities for training and professional development has the project provided?Amruta Bapat is a graduate student in the Interdepartmental Genetics and Genomics program at Iowa State University who was supported full time by this grant. This year, in addition to her normal course work, she participated in the following activities: 1. Attended ASTA CSS 2018 and Seed expo held between Dec 3-6, 2018, Chicago and presented poster titled " A molecular model for thega1locus in maize genotypes". 2. Attended USTN annual meeting on December 4th 2018. 3. Attend the 6th Annual R. F. Baker Plant Breeding Symposium at ISU on 1st March 2019 and presented poster titled " A molecular model for thega1locus in maize genotypes". 4. Attended and presented poster titled "Analysis of a cluster of ZmPME3 pseudogenes at the maize Gametophytic Incompatibility 1 (Ga1-s) locus" at Corn breeding research meeting between March 13-14, 2019 at St. Louis. 5. Attended and presented poster titled " Analysis of a cluster of ZmPME3 pseudogenes at the maize Gametophytic Incompatibility 1 (Ga1-s) locus" at the AnnualMaizeGenetics Conference between March 14-17th 2019 at St. Louis. 6. Participated in the 3-day hands on Plant CRISPR Workshop held between May 13-15, 2019 at Crop Bioengineering center at Iowa State University. Nicholas (Nick) Boerman is an Iowa State University graduate student in the plant breeding program, supported full time by this project. In addition to his course work, he participated in the activities 1. - 3. mentioned above. In addition, he gave an oral presentation at the ICSDA sweet corn meeting in Wisconsin, 2018, and he will have an oral presentation at the ASA/CSSA meeting 2019 in San Antonio. Dr. Anderson Verzegnazzi was a graduate student in the plant breeding program at ISU. Even though not covered by this project, he developed the BS39-derived inbred lines used in this project, and was involved in the 2018 field trials. Anderson defended in July 2019, and is meanwhile working as plant breeder for KWS Saat AG in his home country Brazil. Dr. Tracy trained Ph.D. student Cathleen McCluskey. Cathleen McCluskey is a masters degree candidate in the interdepartmental Agroecology Program at University of Wisconsin-Madison. In this project, she receives training in maize genetics, disease identification, experimental design, statistics, and nursery operation, including planting, pollinating, and harvesting seed. Her thesis work involves measuring and assessing genetic diversity in U.S. maize. Drs. Scott and Lubberstedt hosted two Ph.D. students from Khon Kaen University in Thailand who are working on adding value to specialty corn verities by developing pigmented varieties. Mr. Prakasit Duangpapeng and Ms. Ponsawan Khamphason worked with project members to carry out field operations this summer. They learned about all aspects of field operations including planning, organization, planting, pollinating, data collection, and harvest. How have the results been disseminated to communities of interest?Results were shared with cooperators scientific meetings including the ASTA Corn and Soybean expo (December 4-6, 2018, Chicago, Il) and the Maize Genetics conference (March 14-17, 2019, St. Louis, Mo.) Nick Boerman he gave an oral presentation at the ICSDA sweet corn meeting in Wisconsin, 2018, and he will have an oral presentation at the ASA/CSSA meeting 2019 in San Antonio. A Field Day was held at the ISU Neely-Kinyon Farm in Greenfield, Iowa, on August 20, where the OREI project was presented and discussed with 52 participants. Dr. Scott presented a talk on corn inbreds, hybrids and triple crosses for organic production systems. On November 25, a workshop on the project and the state of organic seed in the U.S. will be held at the Iowa Organic Conference in Iowa City, Iowa. We continue to have buy-in from project farmers to grow the specific varieties or lines, and assist with data collection during the season. A new farmer from SW Iowa will participate in 2020. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Dr. Tracy and Cat McCluskey hosted a group of organic farmers from Germany and the USA. We toured the ISU plots grown in Wisconsin and gave a presentation on this project. They were 19 attendees. Dr. Tracy presented at talk at the UW-Madison organic cropping field day, Arlington Wisconsin August 29, 2019. There were approximately 60 attendees. What do you plan to do during the next reporting period to accomplish the goals?Dr. Lubberstedt: In the 3rd year of the project, we will evaluate testcrosses of BS39 inbred lines under organic field conditions at the ISU Agronomy farm and in Wisconsin (2nd year in both cases). We will continue our introgression program of qSHGD1 and Ga1 into 10 BS39 and 3 sweet corn derived lines by backcrossing. We hope to complete the approval process of the "organic plant breeding" certificate program, and initiate course development for a first offering fall 2020. We will prepare first publications on characterization of BS39 inbred lines, and report findings at different conferences and meetings (ICSDA, stakeholder meeting, MOSES, MGC, Corn Breeder school, ..). Dr. Delate: Hybrids from germplasm developed under the 2014 NIFA - OREI grant, and any germplasm that becomes available from this OREI grant, will be tested in organic farmer-cooperators' fields. Sweetcorn germplasm from UW's program will also be tested. Field Days will be held on at least two on-farm sites in 2020. Dr. Scott: We will continue to study the inheritance of gametophytic incompatibility systems in different families of maize in order to develop a selection strategy that effectively transfers the trait to desired varieties. Our current focus is on understanding genotypic variation in pollen exlusion, and genotype-dependent variation in gene action. We will evaluate hybrids derived from high methionine doubled haploid lines to identify those with superior agronomic traits. Dr. Tracy: We will evaluate the progenies being developed for pollen exclusion and SHGD and sweet corn characteristics. We will also grow locations of the ISU trials and supply sweet corn cultivars to interested participants. We will continue our trials on ways to improve organic production of hybrid corn seed.

    Impacts
    What was accomplished under these goals? Overall impact statement: The overall goal of the project is to substantially increase the productivity of organic field and sweet corn production within organic crop rotations, by improving the efficiency of breeding through doubled haploid (DH) technology and traditional breeding methods, focused on early plant vigor and competitiveness factors. Another goal is to develop a Certificate program in Organic Plant Breeding, which can be accessed online in the U.S. and internationally. All project partners conducted field experiments for evaluation of maize lines under organic conditions, and / or breeding work to develop novel germplasm. Superior lines with regard to agronomic performance under organic conditions and for quality traits were identified. These superior lines will receive by conventional crosses additional properties which will make future breeding of organic maize varieties more efficient. A major finding was that an important property for DH technology is simple inherited (one single major gene), which will simplify breeding strategies to incorporate this property into elite breeding materials. Our findings will lead to changes in knowledge and action in the way breeding of organic field and sweet corn will be done in future, by using DH technology and the novel genetic property characterized in this project. Outcomes will be first generation field and sweet corn lines carrying this property along with another important trait (cross-incompatibility) employed in this project. Stakeholders will be engaged to evaluate first prototypes under organic practise conditions, to determine robustness of this materials. The certificate program on Organic Plant Breeding to be established in this program will be a venue, to share new knowledge in the field or organic plant breeding with students, working professionals, and others engaged in this topic. Strengthened organic plant breeding will lead to better adapted and more competitive varieties under organic conditions, which benefits farmers but also the general public, as this should impact prices of organic products. Objective 1 Field corn, BS39-derived lines: From the tropical BS39 population adapted by Dr. Hallauer (ISU) to Iowa conditions, more than 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD). Large variation for grain yield and other agronomic traits was observed in per se (3 locations) and testcross evaluations (4 locations) in 2018. Besides lines exceeding commercial checks, quite a few lines showed lodging, high kernel moisture at harvest, among others. Only the best 50 lines were evaluated under organic conditions in two locations (Ames, Wisconsin), to be repeated in 2020. In addition, a 2nd cycle breeding approach was adopted: the best two lines with SHGD ability were crossed with the 10 best lines with 100% BS39 background. 2nd generation inbreds with SHGD ability and 75% BS39 genome background, to be evaluated summer 2021. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Development of donor lines in field and sweet corn carrying Ga1 and the major QTL for SHGD by marker-assisted backcrossing is proceeding as planned. In addition, based on mapping information for the 400 BS39-derived inbred lines, a major SHGD QTL was responsible for the high success rate of obtaining DH lines without colchicine. Field and sweet corn, gametophytic incompatibility: To minimize the possibility of GMO contamination, we are developing pollen excluding varieties of corn using a natural pollen exclusion (PE) process called gametophytic incompatibility (GI). We continued to advance sweet corn breeding populations for the purpose of developing pollen excluding sweet corn lines. These lines contain the Ga2 GI locus known to exclude pollen of non-Ga2 varieties, including GMO varieties. This year we carried out tests for PE ability and will advance those plants that display the ability. The extent of PE systems in maize germplasm is poorly understood, making it difficult to plan deployment of these systems. In an effort to better understand the distribution of the three known pollen exclusion systems, we also carried out tests for the ability of sweet corn inbreds to overcome the PE barriers Ga1, Ga2 and Tcb-1. Sweet corn lines were crossed to plants know to possess each of the three PE barriers. In the unlikely event that such crosses do produce kernels, we will characterize PE ability of these sweet corn lines in more detail. Objective 2 We produced hybrid seed to distribute to farmers for observation of two high-yielding hybrids, and produced seed of six experimental hybrids produced from DH parents with high levels of methionine. We produced test cross seed of 22 promising hybrids for evaluation next year and carried out one organic yield trial containing 32 hybrids. This activity will allow our farmer-cooperators to evaluate the products of this grant and to provide feedback to the researchers, ensuring that grant activities are relevant to the needs of our customers. Objective 3 Field plot design and data collection of 2019 farmer trials were finalized at the MOSES conference 2019. The 2019 season was challenging. Cold and wet weather delayed planting by two to three weeks; July's rainfall was 80% below normal; and harvests have been delayed due to wet weather. The trial at ISU Neely-Kinyon Farm in Greenfield, Iowa, was not planted until June 12, where five USDA lines were compared. Emergence was depressed due to the wet weather, averaging 70% in the 5769 and 8517 lines, and significantly lower, at 56%, in the other lines. Yields and grain quality have not yet been calculated from this site due to wet weather delaying harvest. The second set of trials was conducted on three farms-two in Iowa (field corn) and Minnesota (sweet corn). The Minnesota farm suffered from a wet spring and only 50% of the sweet corn plants produced ears. The White Earth Nation 4-H students, who helped maintain the plots, reported high kernel sweetness. The field corn sites, while enduring excessive rains early and late in the season, fared well in 2019.Mid-July commercial organic variety plant height was equivalent across the two on-farm sites, averaging 158 cm. Weed management was more difficult in 2019. Grass weeds averaged 16 weeds/sq. meter across all sites. Broadleaf weeds averaged 5 weeds/sq. meter. The breeder's corn could not be planted on-farm this year, due to the NC site experiencing excessive rains for an entire month. Stalk nitrate samples were collected from two on-farm sites and analyzed by a commercial lab. Stalk nitrate content, which ranged from 345 ppm at the NW farm to 1,331 ppm at the NE farm, demonstrated the lack of nitrate pollution potential usually experienced with conventional corn production. Ear samples were hand-collected from each variety at physiological maturity at two of the three sites and dried to 15% moisture. Corn ear weight averaged 200 g/ear for Blue River 57A30 compared to 218 g/ear for Viking 5014. Kernel weight per ear averaged 168 g for Blue River 57A30 compared to 198 g/ear for Viking 5014. To date, none of the on-farm field corn trials has been machine-harvested for yields because of wet weather. After harvest, we will determine grain quality from ear samples from all sites. Objective 4 The "Organic Plant Breeding" course has been approved by the CALS curriculum committee. A proposal for the "Organic Plant Breeding" certificate program has been developed and approved by the Department of Agronomy curriculum committee and faculty. Next step is approval by the CALS curriculum committee. Once approved, the Certificate program can be offered, as three courses are already available, while the "Organic Plant Breeding" course (as last course in the program) would be developed.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Khamphasan, P., Lomthaisong, K., Harakotr, B., Ketthaisong, D., Willcox, M., Scott, M.P., Lertrat, K., Suriharn, B. 2018. Genotypic variation in anthocyanins, phenolic compounds and antioxidant activity in cob and husk of purple field corn. Agronomy Journal. 8(11):271. https://doi.org/10.3390/agronomy8110271.
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Duangpapeng, P., Ketthaisong, D., Lomthaisong, K., Lertra, K., Willcox, M., Scott, M.P. 2018. Corn tassel: A new source of phytochemicals and antioxidants for value-added products in agro-industry. Agronomy Journal. 8:242. http://dx.doi.org/10.20944/preprints201809.0517.v1.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Delate, K., T. Lubberstedt, P. Scott, W. Tracy and R. Turnbull. 2019. Developing field and sweet corn hybrids for organic producers: The ACES project. ASA Annual Meeting, San Antonio, TX. https://scisoc.confex.com/scisoc/2019am/meetingapp.cgi/Paper/120078
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Boerman, N., Frei, U.K., and T. Lubberstedt. 2019. Characterization of Spontaneous Haploid Genome Doubling in Maize. ASA Annual Meeting, San Antonio, TX. https://scisoc.confex.com/scisoc/2019am/meetingapp.cgi/Paper/118480
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: On-Farm Organic Corn Trials  Farmer-Cooperator Meeting, Midwest Organic and Sustainable Education Service (MOSES) Organic Conference, La Crosse, WI, 2/23/19
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Organic Research in Iowa - Horticulture Dept., University of Florida, Gainesville, FL, 3/20/19
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Organic Research Update  OREI ACES Project  ISU Neely-Kinyon Farm Field Day, Greenfield, IA, 8/20/19, K. Delate and Paul Scott
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Iowa Organic Research Update  Oregon State University, Corvallis, OR, 10/24/19
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Delate, K. 2019. Organic Field Day Planned at Neely-Kinyon Farm, ISU Extension and Outreach, Ames, IA: July 31, 2019: https://www.extension.iastate.edu/news/organic-field-day-planned-neely-kinyon-research-farm
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Sanchez, D., Liu, S., Ibrahim, R., Blanco, M., L�bberstedt, T. (2018) Association mapping of seedling root traits in exotic derived doubled haploid lines of maize. Plant Science 268:30-38
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Andorf, C., Beavis, W.D., Hufford, M., Smith, S., Suza, W., Wang, K., Woodhouse, M., Yu, J., L�bberstedt, T. (2019) Technological advances in maize breeding: Past, present and future. Theor. Appl. Genet. 132: 817-849
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Yang, J., Li, H., Qu, Y., Chen, Q., Tang, J., L�bberstedt, T., Liu, Z. (2019) Identification of QTL for haploid male fertility in maize (Zea mays L.). Plant Breeding 138: 259-265
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Vanous, A., Blanco, M., Martin-Schwarze, A., Flint-Garcia, S., Bohn, M., Edwards, J., L�bberstedt, T., Gardner, C. (2019) Stability analysis of kernel quality traits in exotic derived maize doubled haploid lines. The Plant Genome 12(1):1-14
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Vanous, K., Jubery, T.Z., Ganapathysubramanian, B., L�bberstedt, T. (2019) A scalable reusable framework for early discrimination of maize haploids. The Plant Phenotype Journal 2:1 DOI: 10.2135/tppj2018.10.0008


    Progress 09/01/17 to 08/31/18

    Outputs
    Target Audience:Organic farmers, Organic organizations, Plant breeders, Seed producers, Seed processors Changes/Problems:Objective 4 is delayed. While a syllabus for the new course on "Organic Plant Breeding" is in place, and its approval ongoing, the time needed for the administrative process to (a) get this new course approved, and (b) the certificate program, was underestimated. We currently plan to offer this course for a first time spring 2020, which is 6 months later than mentioned in the project description. On the positive side, we gained approval for development and offering this course far beyond duration of the ongoing project. Thus, once developed it will be available through the distance education in plant breeding platform at ISU. What opportunities for training and professional development has the project provided?Dr. Lubberstedt trained Ph.D. student Nicholas Boerman.Nicholas (Nick) Boerman is an Iowa State University graduate student in the plant breeding program. Nick received his M.Sc. degree in plant breeding at Texas A & M University, where he took fundamental courses in plant breeding, genetics and statistics. At ISU, he is taking more advanced classes on these topics, as well as on molecular genetics and genomics and bioinformatics. In addition, he participated in the R.F. Baker Symposium (Plant Breeding) at ISU in March 2018, and attended a summer workshop on Python and R programming this summer (in NSF P3 proram). Nick's research addresses doubled haploid (DH) technology, specifically spontaneous haploid genome doubling, and its introgression into field and sweet corn. Nick has been exposed to all steps of DH technology as part of his field nursery work, also attended a DH workshop at ISU this summer. Nick prepared a research proposal, which he presented to his program of study committee. He already passed two out of five written prelims, aiming for completing all prelims within the next year. Dr. Scott trained Ph.D. student Amruta Bapat and undergraduate student Taylor Hintch. Taylor Hintch is an Iowa State University undergraduate student majoring in Global Resource Systems. As an hourly worker in this project, she receives training in corn genetics, participating in all aspects of the nursery operation, including planting, pollinating, harvesting, and inventorying seed. In addition, she has her own research project that involves characterization of a series of doubled haploid maize lines with enhanced nutritional quality. In this project, she learned fundamentals of experimental design and laboratory methods for evaluating nutritional quality. She prepared a poster and presented her results at the Borlaug Lecture poster contest at Iowa State University, where she won first prize in the Undergraduate division. Amruta Bapat is an Iowa State University graduate student in the Interdepartmental genetics and genomics graduate program. She receives training in maize genetics and molecular biology by assisting with shared nursery operations and carrying out her thesis research. Her thesis project involves understanding the molecular mechanism controlling pollen exclusion in the Ga1 genetic system. In this research she has learned bioinformatics analysis, and molecular biology methods such as DNA manipulations and heterologous gene expression. Dr. Tracy trained Ph.D. student Cathleen McCluskey. Cathleen McCluskey is a masters degree candidate in the interdepartmental Agroecology Program at University of Wisconsin-Madison. In this project, she receives training in maize genetics, disease identification, experimental design, statistics, and nursery operation, including planting, pollinating, and harvesting seed. Her thesis work involves measuring and assessing genetic diversity in U.S. maize. How have the results been disseminated to communities of interest?A first stakeholder meeting with representatives from plant breeding companies, seed producers, seed processors, and organic organization was held at the beginning of the USDA OREI ACES project in December 2017, in conjunction with the ASTA meeting in Chicago. Project co-PIs presented the planned activities within the four project objectives, which were subsequently critically discussed. A follow-up meeting is planned December 2018 in conjunction with ASTA, combined with a meeting of the US testing network (USTN). One of the most critical aspects of this component of ACES was the buy-in from project farmers to maintain the variety trial fields in excellent condition throughout the season and their assistance with data collection. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Taylor Hintch presented a poster titled "Analyzing doubled haploids using a microbial assay to identify high methionine lines" at the Borlaug Lecture Poster Session (Ames, Iowa, October 15, 2018) and won first prize in the undergraduate poster contest. A report of the sweet corn portion will be given at the international sweet corn development association annual meeting in December. What do you plan to do during the next reporting period to accomplish the goals?Dr. Lubberstedt: In the 2nd year of the project, we will evaluate testcrosses of BS39 inbred lines under organic field conditions at the ISU Agronomy farm. We will continue our introgression program of qSHGD1 and Ga1 into 10 BS39 and 3 sweet corn-derived lines by backcrossing. We hope to complete the approval process of the "organic plant breeding" course and respective certificate program, and initiate course development for a first offering spring 2020. We will prepare first publications on characterization of BS39 inbred lines, and report findings at different conferences and meetings (ICSDA, stakeholder meeting, PAG, MOSES, MGC, Corn Breeder school, ..). Dr. Delate: Hybrids from germplasm developed under the 2014 NIFA - OREI grant, and any germplasm that becomes available from this OREI grant, will be tested in organic farmer-cooperators' fields. Sweetcorn germplasm from UW's program will also be tested. Field Days will be held on at least two on-farm sites in 2019. Dr. Scott: We will continue to study the inheritance of gametophytic incompatibility systems in different families of maize in order to develop a selection strategy that effectively transfers the trait to desired varieties. Our current focus is on understanding genotypic variation in pollen exlusion, and genotype-dependent variation in gene action. We will evaluate hybrids derived from high methionine doubled haploid lines to identify those with superior agronomic traits. Dr. Tracy: We will continue to work with the over 50 unique crosses between sweet corn genotypes and Ga1 or Ga2. These crosses have been sent to winter nurseries where they will be self pollinated generating 50 F2 populations from which we will derive new inbreds.

    Impacts
    What was accomplished under these goals? Overall impact statement: The overall goal of the project is to substantially increase the productivity of organic field and sweet corn production within organic crop rotations, by improving the efficiency of breeding through doubled haploid (DH) technology and traditional breeding methods, focused on early plant vigor and competitiveness factors. Another goal is to develop a Certificate program in Organic Plant Breeding, which can be accessed online in the U.S. and internationally. All project partners conducted field experiments for evaluation of maize lines under organic conditions, and / or breeding work to develop novel germplasm. For example, 400 lines developed for this project were evaluated in six environments. Superior lines with regard to agronomic performance under organic conditions and for quality traits were identified. These superior lines will receive by conventional crosses additional properties which will make future breeding of organic maize varieties more efficient. A major finding was that an important property for DH technology is simple inherited (one single major gene), which will simplify breeding strategies to incorporate this property into elite breeding materials. Our findings will lead to changes in knowledge and action in the way breeding of organic field and sweet corn will be done in future, by using DH technology and the novel genetic property characterized in this project. Outcomes will be first generation field and sweet corn lines carrying this property along with another important trait (cross-incompatibility) employed in this project. Stakeholders will be engaged to evaluate first prototypes under organic practise conditions, to determine robustness of this materials. The certificate program on Organic Plant Breeding to be estbalished in this program will be a venue, to share new knowledge in the field or organic plant breeding with students, working professionals, and others engaged in this topic. Strengthened organic plant breeding will lead to better adapted and more competitive varieties under organic conditions, which benefits farmers but also the general public, as this should impact prices of organic products. Objective 1 Develop novel organic field corn and sweet corn genotypes for the commercialization pipeline that utilize more efficient methods of breeding. Field corn, BS39-derived lines: From the tropical BS39 population that was adapted by Dr. Hallauer (ISU) to Iowa conditions, 400 inbred lines were derived by the single seed descent (SSD) or doubled haploid (DH) approach, the latter using either colchicine for genome doubling, or a colchicine-free genetic mechanism for spontaneous haploid genome doubling (SHGD). These 400 lines were evaluated at per se level in three environments in Iowa, including one organic location at the ISU Agronomy farm. Moreover, testcross trials with those same 400 lines were conducted under conventional conditions (4 locations), and will be conducted under organic conditions at the ISU Agronomy farm in 2019, and 2020. All field BS39-experiments for 2018 have been completed and harvested. Respective data are under analysis and will be presented at the stakeholder meeting in December 2018. Field corn, quality protein maize: Methionine is an important nutrient in the diets of poultry. Most poultry diets in the US are based on corn, and synthetic methionine is added to meet the nutritional needs of the birds. Organic standards limit the level of synthetic methionine in poultry diets, so corn with levels of methionine that are high enough to offset the need for supplementation are desirable. High methionine field corn lines produced by doubled haploid technology were evaluated per se for their ability to produce high grain methionine levels. Five lines were identified that had significantly higher methionine levels than the high methionine inbred check. These lines have the highest level of methionine of any inbred observed to date. The lines with the highest levels of methionine were used to make hybrid seed that will be evaluated in yield trials next year. Hybrids of high methionine lines and commercial inbreds were observed for the purpose of assigning them to heterotic groups. We concluded that these inbreds combine equally well with stiff stalk and non-stiff stalk commercial inbreds, making them desirable for use as hybrid parents. Introduction of SHGD and Ga1 into field and sweet corn (ISU): Genetic mapping experiments at ISU revealed, that a single major QTL (qSHGD1) explains more than 50% of the phenotypic variation for SHGD. A donor line fixed for qSHGD1 and cross-incompatibility gene Ga1 for pollen control has been completed. Moreover, crosses and marker-assisted backcrosses have been initiated to develop 10 BS39- and 3 sweet corn-derived lines carrying both qSHGD1 and Ga1. Dr. Tracy evaluated in a four replication inbred trial selected BS39 lines that were chosen for this conversion program under organic conditions at West Madison WI. He supplied performance data to the originators at ISU. Sweet corn (Wisconsin): We have made over 50 unique crosses between sweet corn genotypes and Ga1 or Ga2. These crosses have been sent to winter nurseries where they will be self pollinated generating 50 F2 populations from which we will derive new inbreds. Objective 2. Select and test on-farm genotypes that meet the critical needs of organic producers. On farm trials for field corn: There were two sets of variety trials with farmer involvement in 2018. Field plot design and data collection were finalized at a project farmer meeting at the MOSES conference on February 23, 2018. The first trial was at the ISU Neely-Kinyon Farm in Greenfield, Iowa (southwest Iowa), where four USDA lines were compared to the native 'Meskwaki Tama Flint' (MTF) corn, which was secured from a member of the Meskwaki Nation. The second set of trials was conducted on five farms-four in Iowa (field corn) and one in Minnesota (sweet corn). The USDA lines, averaging 123 bu/acre, with the 393 line more competitive at 138 bu/acre, compared well with the commercial organic variety (Viking .79-08, Albert Lea, MN), which yielded 173 bu/acre. We are currently calculating grain quality from ear samples from all sites. Organic producers need corn varieties that differ from those used in conventional agriculture systems. They must be GMO-free, have native resistance to insect pests and pathogens, and compete effectively with weeds. Hybrids with traits desired by organic producers were identified from germplasm developed under a 2014 NIFA - OREI grant. Seed of these hybrids was produced in sufficient quantity to share with farmers next year. Objective 3. Practice technology transfer through a network with organic farmers and organic seed companies. One of the most critical aspects of this component of ACES was the buy-in from project farmers to maintain the variety trial fields in excellent condition throughout the season and their assistance with data collection. This shows great promise for their continued involvement in evaluating new lines that are developed from the ACES pipeline. Dr. Tracy gave a field tour to a group from one of the major organic seed sales companies and we have also carried out trials with major organic hybrid seed producer. Objective 4. Develop a graduate Certificate program in Organic Plant Breeding. A syllabus for the novel course "Organic Plant Breeding" has been established. The course is currently routing in the ISU approval process. As important initial step, the head of Department of Agronomy approved development and continuous offering of this course, which requires resources beyond this ongoing USDA OREI project ACES. Currently, approval is requested from the College of Agriculture and Life Science curriculum committee. Once the approval process is completed, actual course development will commence, targeted for a first offering spring 2020.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2018 Citation: De la Fuente, G., Frei, U.K., Trampe, B., Nettleton, D., L�bberstedt, T. (2018) A diallel analysis of a maize donor population response to in vivo maternal haploid induction. I: Inducibility. Crop Sci. 58:1830-1837
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: De La Fuente, G.N., Carstensen, J.M., Edberg, M.A., L�bberstedt, T. (2017) Discrimination of haploid and diploid maize kernels via multispectral imaging. Plant Breeding 136:50-60
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wu, P., Ren, J., Tian, X., Li, G., Li, W., L�bberstedt, T.,Wang, L., Liu, W., Chen, S. (2017) New insights into the genetics of haploid male fertility. Crop Sci. 57:637-647
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Vanous, K., Vanous, A., Frei, U.K., L�bberstedt, T. (2017) Generation of maize (Zea mays) doubled haploids via traditional methods. Current Protocols in Plant Biology 2: 147-157
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wang, C., Hu, S., Gardner, C., L�bberstedt, T. (2017) Emerging avenues for utilization of exotic germplasm. TIPS 22: 624-637
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Ren, J., Wu, P., Tian, X., L�bberstedt, T., Chen, S. (2017) Fine mapping of quantitative trait locus qhmf4 causing haploid male fertility in maize based on segregation distortion. Theor. Appl. Genet. 130:13491359
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Ren, J., Wu, P., Trampe, B. , L�bberstedt, T., Chen, S. (2017) Novel technologies in doubled haploid line development. Plant Biotech J. 15: 1361-1370
    • Type: Conference Papers and Presentations Status: Other Year Published: 2018 Citation: On-Farm Organic Corn Trials  Farmer-Cooperator Meeting, Midwest Organic and Sustainable Education Service (MOSES) Organic Conference, La Crosse, WI, 2/24/18
    • Type: Conference Papers and Presentations Status: Other Year Published: 2018 Citation: Organic Research in Iowa - Horticulture Dept., University of Florida, Gainesville, FL, 3/14/18
    • Type: Conference Papers and Presentations Status: Other Year Published: 2018 Citation: Organic Research Update  OREI ACES Project  ISU Neely-Kinyon Farm Field Day, Greenfield, IA, 8/12/18, K. Delate and Paul Scott
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Vanous, A., Gardner, C., Blanco, M., Martin-Schwarze, A., Flint-Garcia, S., Bohn, M., Edwards, J., L�bberstedt, T. (2018) Association mapping of flowering and plant height traits in germplasm enhancement of maize doubled haploid (GEM-DH) lines. The Plant Genome 11:170083