Source: WASHINGTON STATE UNIVERSITY submitted to
FRAME: FUNGICIDE RESISTANCE ASSESSMENT, MITIGATION AND EXTENSION NETWORK FOR WINE, TABLE, AND RAISIN GRAPES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1016359
Grant No.
2018-51181-28364
Cumulative Award Amt.
$4,743,734.00
Proposal No.
2018-03375
Multistate No.
(N/A)
Project Start Date
Sep 1, 2018
Project End Date
Aug 31, 2023
Grant Year
2019
Program Code
[SCRI]- Specialty Crop Research Initiative
Project Director
Moyer, M.
Recipient Organization
WASHINGTON STATE UNIVERSITY
240 FRENCH ADMINISTRATION BLDG
PULLMAN,WA 99164-0001
Performing Department
Agricultural Research Center
Non Technical Summary
In specialty crops, product quality is king. What happens when routine production tactics that influence quality, such as disease management, stop working? This situation has become reality in grape (wine, table, raisin) production in the USA, where control failures of powdery mildew (Erysiphe necator) have recently occurred. Many factors can influence the effectiveness of a disease management program, but fungicide resistance is the most feared. There is currently no effective system to monitor or predict fungicide resistance; it is usually identified after a management failure. Our proposed research and extension efforts will empower growers with data and predictive tools on the potential for fungicide resistance development that will be coupled with improved approaches to managing and mitigating resistance development, allowing them to design and implement fungicide resistance stewardship programs. We will develop rapid monitoring technology and the implementation programs for diagnostic labs. Our systems-approach will enable all audiences (e.g., growers, consultants, extension, chemical manufacturers and resellers) to engage in developing stewardship programs to protect at-risk fungicides. The integration of this project's objectives address: Where fungicide resistance is currently; Improve how we detect and monitor fungicide resistance and improve application efficiency; Predict where and when fungicide resistance will arise; and Develop strategies that help growers, educators, and manufactures answer what they need to do to mitigate resistance development and manage resistance that has already developed. Importantly, this Fungicide Resistance Assessment, Mitigation and Extension Network (FRAME Network) will develop tools that can be applied to other specialty crops facing fungicide resistance challenges.
Animal Health Component
45%
Research Effort Categories
Basic
45%
Applied
45%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2124020106060%
2121139106040%
Goals / Objectives
In specialty crops, product quality is king. What happens when routine production tactics that influence quality, such as disease management, stop working? This situation has become reality in grape (wine, table, raisin) production in the USA, where control failures of powdery mildew (Erysiphe necator) have recently occurred. Many factors can influence the effectiveness of a disease management program, but fungicide resistance is the most feared. There is currently no effective system to monitor or predict fungicide resistance; it is usually identified after a management failure. Our proposed research and extension efforts will empower growers with data and predictive tools on the potential for fungicide resistance development that will be coupled with improved approaches to managing and mitigating resistance development, allowing them to design and implement fungicide resistance stewardship programs. We will develop rapid monitoring technology and the implementation programs for diagnostic labs. Our systems-approach will enable all audiences (e.g., growers, consultants, extension, chemical manufacturers and resellers) to engage in developing stewardship programs to protect at-risk fungicides. The integration of this project's objectives address: Where fungicide resistance is currently; Improve how we detect and monitor fungicide resistance and improve application efficiency; Predict where and when fungicide resistance will arise; and Develop strategies that help growers, educators, and manufactures answer what they need to do to mitigate resistance development and manage resistance that has already developed. Importantly, this Fungicide Resistance Assessment, Mitigation and Extension Network (FRAME Network) will develop tools that can be applied to other specialty crops facing fungicide resistance challenges.Objective 1: Extension and OutreachObj. 1 - Activity 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United StatesObj. 1 - Activity 2: In-the-field demonstrations of management recommendations.Obj. 1 - Activity 3: Develop a "University Service Center" business plan for fungicide resistance testing.Obj. 1 - Activity 4: Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Objective 2: Group DecisionsObj. 2 - Activity 1: Conduct interviews to identify the externalities negotiating parties are experiencing (i.e., how their pesticide use affects their neighbors' production).Obj. 2 - Activity 2: Use game theory to develop compensating mechanisms that induce cooperation and aid fungicide stewardship by inducing cooperative behavior among growers, crop consultants and chemical companies.Objective 3: Detection and MonitoringSub-Objective 3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection.Obj. 3.1 - Activity 1: Development of PCR based diagnostic assays.Obj. 3.1- Activity 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance.Obj. 3.1 - Activity 3: Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Sub-Objective 3.2 - Monitoring for Fungicide Resistance.Obj. 3.2 - Activity 1: Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Obj. 3.2 - Activity 2: Phenotype for fungicide sensitivity.Sub-Objective 3.3 - Understanding the Basis of Resistance.Obj. 3.3 - Activity 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides.Obj. 3.3 - Activity 2: Create reference genome of E. necator for comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Obj. 3.3 - Activity 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis.Objective 4: Predicting Resistance SpreadSub-Objective 4.1: Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity.Obj. 4.1 - Activity 1: Sample collections and phenotyping.Obj. 4.1 - Activity 2: Genome sequencing and allelic diversity analyses.Sub-Objective 4.2 - Dispersion Tracking and Population Prediction.Obj. 4.2 - Activity 1.1: Mapping resistant isolate spread risk.Obj. 4.2 - Activity 1.2: System validation and evaluation.Sub-Objective 4.3 - Fungicide Record Evaluation.
Project Methods
Objective 1: Extension and OutreachHypotheses: (1) Ready-to-use extension and outreach information will improve implementation and use of recommendations; and (2) Providing in-field examples of resistance mitigation techniques will improve grower adoption of proposed management recommendationsEffortsNational survey to develop baseline knowledge; change in knowledge.In-the-field demonstrations of management recommendations.Create and engage with enhanced local industry groups.Develop train-the-trainer workshops and classic extension products on fungicide resistance management.Develop a "Service Center" business plan for resistance testing.EvaluationMitigation efficacy determined by comparing resistance frequency and tracking crop loss in vineyards using modified programs.National survey in yrs 1 and 4 to determine changes in stakeholder knowledge/behavior.Tracking downloads and web hits for Extension presentations, curricula.MetricsShort-Term: Extension curricula improve grower knowledge on resistance fungicide management knowledge.Mid-Term: Wide-spread adoption of routine resistance testing.Long-Term: Reduction in economic loss due to fungicide resistance; reduction in overall pesticide use as growers stop using "rescue" sprays.Objective 2: Group DecisionsHypotheses: (1) Conflicting interests can be solved through negotiation between growers and chemical companies and; (2) Providing a compensating mechanism will induce cooperation among agents involved in the negotiation.EffortsInterviews to identify the source of conflict among the negotiating parties.Game theory simulation to develop compensating mechanisms that induce cooperation.Policy evaluations.EvaluationDiscussion with growers about the result of the simulation and identification of scenarios that represent their current negotiation problem.MetricsShort-Term: Identification of the source of conflict during the bargaining process.Mid-Term: Characterization and simulation of the bargaining process among growers.Long-Term: Policies are developed that promote cooperation towards resistance management.Objective 3: Detection and MonitoringHypotheses: (3.1) Field-level diagnostics based on nucleic acid amplification will be suitable for detecting presence of fungicide resistance. (3.2) Rapid sampling protocols can be used in place of intensive inoculum monitoring to accurately assess fungicide resistance presence and make management decisions. (3.3) Direct sequencing of genes or transcripts encoding fungicide resistance targets will allow for detection of low-frequency resistant genotypes before practical resistance is observed.Efforts3.1 - Developing Molecular Markers to Improve the Speed of Resistance DetectionDevelopment of PCR based diagnostic assays.Developing isothermal diagnostic tools for in-field detection of fungicide resistance.Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.3.2 - Monitoring for Fungicide Resistance.Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Phenotype for fungicide sensitivity.Collect spatial and temporal data on fungicide resistance for Objective 4.3.3 - Understanding the Basis of Resistance.Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHIs and AzN.Create reference genome of E. necator for comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis.EvaluationSampling methods must meet growers' expectations for ease of collection while balancing sensitivity of detection.Surveys will be conducted that assess grower probability of using different collection methods.Adoption of diagnostic assays by private labs and crop consultants.Implementation of in-field assays with growers and crop consultants.MetricsShort-Term: New tools become available to help diagnostic labs identify potential fungicide resistance before field failures occur.Mid-Term: Identify diversity of mutations associated with FRAC 3, 7 and 13 fungicide resistance for new diagnostic assay developmentLong-Term: Fungicide resistance monitoring becomes routine practices in vineyard management plans.Objective 4: Predicting Resistance SpreadHypotheses: (4.1) Sub-regions, or even vineyards, will have different genotypes and fungicide resistance allele frequencies. (4.2) Highly-resolved biophysical models for air turbulence and vineyard microclimate can be used to significantly improve the prediction of regional spread and risk of fungicide resistant E. necator. (4.3) Fungicide resistance development can be predicted by analysis of historical use.Efforts4.1 - Seasonal Monitoring and Recurring Collection of IsolatesEngage with growers to Identify vineyards for recurring isolate collection.Collect isolates for DNA extractions; monospore isolates for DNA sequencing.Phenotype subset of isolates for resistance to FRAC chemistries 3,7,11,13.Collect and analyze next generation sequencing data4.2 - Dispersion Tracking and Population PredictionConstruct FRAME dispersion risk system.Quantify FRAME dispersion risk system uncertainty (and reduce if possible).4.3 - Fungicide Record EvaluationSolicit historical fungicide programs from growers to determine use pattern influence on resistance.EvaluationValidated genetic features are used to develop diagnostic detection tools.Uncertainty of risk maps is bounded and of reasonable magnitude.Grower adoption rate of fungicide use histories in developing immediate and long-term fungicide plans.MetricsShort-Term: Identification of key features from historical fungicide use that may lead to development of resistance; use of that knowledge to improve spray program design.Mid-Term: Use of population models (genetic, dispersion) to predict where fungicide resistant isolates may spread. Understand how fungicide resistant-isolates spread, and how to predict that spread.Long-Term: Policies of sales and distribution of at-risk fungicides are based on risk-data and follow multi-year approaches to mitigate practical resistance development.

Progress 09/01/18 to 08/09/23

Outputs
Target Audience:Our target audience consists of a diverse body of stakeholders including but not limited to university and government scientists and extension specialists, grape growers, vineyard managers, farm consultants, vineyard management businesses, state and federal agencies focused on the use of agrochemicals, pesticide policy makers and agents, and fungicide retailers. Whether they make, sell, use, recommend, or regulate fungicides, all parties play an important role in fungicide stewardship and the maintenance of effective tools for vineyard disease management. We engaged with these audiences through extension activities (workshops, seminars, virtual presentations), developing new diagnostic tools to detect fungicide resistance, providing information on how to use those tools, and conducting survey work to better understand the underlying factors behind social decisions around fungicide resistance development and management. Over the life of the project, we have presented at various local, regional, national and international scientific and grower meetings, including: in-person and virtual extension workshops and grower meetings across the United States, the American Phytopathological Society annual meeting, the International Grapevine Downy and Powdery Mildew workshop, as well as international agrichemical conferences. We have also used alternative media methods and materials (newsletters, handouts, and trade publications), to create durable resources that our audience can access long after the end of a grant-funded project. We have specifically targeted grower groups and organizations, as evidence-based powdery mildew and fungicide management practices can improve or maintain yields (fewer losses to disease) and produce high quality fruit to meet internal, contract and winery standards. Proper fungicide stewardship and responsible management programs also minimize economic and environmental impacts, including but not limited to fewer tractor passes in the vineyard, worker protection, and reduced carbon footprint for transport and application of fungicides. Indirectly, through peer-to-peer interactions also the project benefits the wider community by providing stability in the agricultural industry, access to a wide range of technical and professional job opportunities, revenue generated from agritourism, habitat for wildlife species and climate resiliency through protection of natural resources and open space. Additionally, responsible pesticide use reduces the risk of exposure and indirect effects of excessive or ineffective applications. Changes/Problems:This project, which ran from September 2019 until August 2023, was significantly impacted by the COVID-19 Pandemic. Exacerbating these challenges was the patchwork differences in allowed and restricted activities, that varied state-to-state, and institution-to-institution. Laboratory Research: Work-from-home, and personnel restrictions in 2020 and 2021 severely limited our ability to access key facilities to complete much of our molecular work. However, advances in genetic technology in this same timeframe has allowed to reach a similar endpoint after our no-cost extension. Many of the scientists were also very creative during the pandemic, as it relates to employee scheduling within acceptable limits of institutional HR rules, to keep the progress moving forward. In some cases, this mean lab activities 24 hours a day, 7 days a week, to accommodate for the 1-person-at-a-time rules that were in place at many institutions. Field Research: Issues with supply chain management that resulted from the pandemic also delayed access to protective gear (facemasks, Tyvek suits), that initially limited our ability to safely work in vineyards and with pesticides. These challenges also lasted for about 2 years but have since been alleviated. Interestingly, the Glove Swab technique we deployed in 2019 for E. necator collection is also the same technique that was adopted for COVID-19 detection, which resulted in some supply bottlenecks during our summer field seasons. Extension Workshops: Our Extension efforts were also initially halted by the pandemic - we had one workshop shut down in the middle of the event (March 2020). Fortunately, our Extension team was able to quickly pivot, and we opted to record several training modules and offer hybrid workshop options - where participants watched training videos online at their leisure, and then met for a ZOOM workshop complete with guided breakout room activities. This allowed us to reach growers all across the United States, and offer more workshops than we had originally anticipated, at a much lower cost to the project. It also changed the way we are planning future Extension events. Institutional Hiring Restrictions: Another major struggle in the last 5 years has been the timely hiring of project personnel. Whether hiring freezes within the USDA-ARS, or hiring freezes and delays at Universities, it is getting more difficult to hire people into technician, project manager, and postdoctoral positions. In many cases, it takes up to a year from the start of the process until someone is in place, and many institutions (including the USDA) are not allowing programs to begin those processes until funding is solidly in place. This significantly impacted expenditure rates, as people were often the largest component to any institutional budget. In this project, by the time we were fully staffed across most institutions, the Pandemic started, which created challenges and delays in training new research assistants in proper sampling techniques, which affected sample quality in some instances leading to unexpected outcomes. What opportunities for training and professional development has the project provided?High School and Undergraduate Students: Several undergraduate and high school students have been engaged in project activities in programs across the country. Most of these opportunities focused on laboratory activities, where students learned basic microbial culturing techniques, as well as greenhouse plant care. Students were also involved in instrument development and deployment as a part of our engineering studies. Graduate Students: This project has provided direct, or indirect, training for a number of graduate students. This includes opportunities to work on FRAME-associated research projects (from field scouting to genome sequencing), to presenting their results to the FRAME scientific and stakeholder advisory groups. Opportunities were also offered to students to participate in writing extension and trade magazine articles (see Products), as well as present their research at relevant scientific conferences (e.g., American Phytopathological Society, and International Grapevine Downy and Powdery Mildew Workshop) or at regional Extension field days. We also offered several opportunities for cross-lab training, which expanded the interdisciplinary activities of FRAME-associated graduates. Graduate students across the USA, but not necessarily associated with FRAME, were engaged as group leaders in our FRAME-specific workshops, to provide an opportunity for Extension engagement and leadership while simultaneously learning from regional grape growers. Postdocs: Multiple postdoctoral scientists have worked full or part-time on FRAME. They received additional field and laboratory training, as well as were provided opportunities to present at our FRAME annual project meetings. In addition, postdocs were engaged in both scientific and Extension publication writing. In addition, our Project Manager position was a postdoctoral position, where the individual also learned about budgeting, meeting coordination, and project management from the Project Director. Technicians: Technicians participated in cross-lab training to expand out diagnostic and PCR activities across the country. They participated in FRAME annual project meetings, were included on the development of scientific posters and papers, and also participated in FRAME Extension workshops. These provided opportunities for professional growth, particularly in scientific communication. Extension Agents and Crop Consultants: Several train-the-trainer style workshops have been offered regionally (i.e., GA), or through FRAME, targeting individuals who assist or consult for grape producers. These workshops have focused on sprayer calibration and spray program design and development, similar to what has been offered to grape growers. However, these consultant workshops went more in-depth on resistance mechanisms and pathogen biology to provide a more in-depth learning and discussion point for participants. Growers: In all of our FRAME-related workshops, we emphasize hands-on learning. As such, participating growers have been trained on sprayer calibration, how to read pesticide labels, how to use available resistance mitigation resources, spray program design, and we also touched on spray program evaluation. The intent was to help build grower confidence in their ability to assess the "fungicide stewardship" aspect of their pest management approaches, rather than just solely focusing on end-efficacy. Stakeholders learned about incidence of fungicide resistance by accessing the Dashboard. How have the results been disseminated to communities of interest?As seen in our list of products, over the life of this project we have delivered: 17 FRAME-associated grower training workshops; 3 podcasts; 27 newsletters, blogs, videos and trade articles; 175 grower or scientific conference presentations; 33 grower or scientific conference posters; 35 peer-reviewed scientific publications (published, in-press, or are currently under review); data and data analysis script deposits into public repositories (NCBI, GenBank, GitHub), 1 project website (routinely updated); 1 fungicide resistance tracking dashboard; and 2 project-associated social media platforms in which we shared the aforementioned products (Twitter/X, Facebook). Conference presentations have ranged from regional and national grower organization meetings, to national and international scientific conferences and symposia. Given our broad targeted audience (students, growers, technicians, scientists, extension specialists, diagnosticians, chemical companies, and consultants), we needed an equally diverse approach to disseminating information. While the "push" approaches (presentations, and social media) helped us to rapidly spread information, we also heavily relied on our project website (framenetworks.wsu.edu), using it as a single source of information on all things FRAME. The website was also where we housed more durable educational products that end-users could continually reference. This sustained approach to developing content and delivering it to varied targeted audiences, allowed FRAME to build "brand" recognition over the course of the project - growers to extension specialists to chemical company representatives in the grape sector across the USA recognize FRAME as an "organization" that is not only developing tools for fungicide resistance diagnostics, but as a credible and reliable source of best-practice information for mitigating crop loss related to fungicide resistance development. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Obj 1: Extension & Outreach Activities: We conducted a national early-project survey that summarized grower's "state of knowledge". We learned that growers understand the basic principles of fungicide resistance, but understanding was influenced by their stage of career. We conducted regional field demonstrations of different powdery mildew management programs in the presence of QoI and DMI resistant E. necator populations, demonstrating the reintegration of QoI fungicides into commercial production systems is feasible without loss under low-pressure conditions. We also demonstrated that the integration of sulfur tank mix or rotation significantly reduces QoI resistance within a E. necator population. We developed a budget models for University and private company service centers to assist them in adoption of our molecular diagnostic techniques. We developed and delivered regional and national workshops on fungicide stewardship, where participants learned about pathogen biology and resistance management concepts and had opportunities for interactive fungicide evaluation sessions. We developed a fungicide resistance dashboard to summarize and display fungicide resistance data in real-time. Impacts: Through this project's efforts, growers identified an improvement in how they read and understand fungicide labels, how they approach spray program design and fungicide rotation, and how the role that pathogen biology and spray application techniques factors into fungicide stewardship. Our molecular diagnostic assays were quickly adopted by commercial testing services, extending the availability of this tool across the country. Growers (>60%) participating in our FRAME-specific reported that they would be changing the way they approach spray program design after attending a FRAME workshop. The dashboard was a communication tool for growers in peer-learning groups, who used the real-time data to advocate for the adoption of fungicide stewardship practices. Obj. 2: Group Decisions Activities: Through surveys, we gathered information about grape growers' willingness to cooperate to manage and mitigate fungicide resistance. The results of the survey indicated that growers are willing to adjust their fungicide use practices if it assisted with the mitigation of resistance. However, they were less willingness to change their practices when that adjustment would negatively impact their profits. In addition, we examine the role of misinformation about fungicide resistance in a compensation mechanism that induces grape growers to reduce their use of fungicide. We found that misinformation (pessimistic or optimistic beliefs) about fungicide resistance can affect the use of fungicides. Impacts: The impacts here are largely targeted related to what can be done by those who influence policies (regulations). Within the US grape communities, cooperation towards mitigating fungicide resistance can be increased by offering a compensation tool of some form. We also learned about the importance of information transmission and how the management of information can help ameliorate the negative effects of fungicide resistance. Obj. 3: Detection & Monitoring Activities: We developed a CYP51-Y136F qPCR assay for detecting Y136F mutation and copy number of the CYP51 gene that was sensitive to 20 conidia and 100% accurate in estimating the genotype of individual isolates. We developed a germination bioassay for FRAC 9, 11, 13, 50 and U6 fungicides that improved throughput by >400%. We developed a novel SNP assay that can detect QoI resistance rapidly in the field. We also identified differentially expressed genes associated with FRAC 13 fungicide resistance in grapes. Across most of the USA, we processed approximately 16,000 E. necator samples for the presence of the G143A and CYP51 mutations, and also identified isolates resistant to FRAC 7 and FRAC 13 fungicides. We demonstrated that there is not complete cross-resistance among different FRAC 3. We also produced a high-quality chromosome-scale 81.1 Mb genome assembly (98% complete; assembled into 11 complete chromosomes) and annotation of E. necator isolate EnFRAME01. We used mutations in SDHB/C/D subunits of E. necator to assess resistance in over 1,000 isolates across US and Canada against SDHI fungicides. In total, 17 missense mutations were identified in one of the SDHI subunits. We determined that SDHI resistance is mediated in E. necator by both target-site resistance and non-target site resistance mechanisms. Impacts: We determined that the QoI qPCR assay was sensitive to a single conidium, 100% accurate, and was a suitable replacement for the En qPCR assay used to monitoring concentration of E. necator in airborne sample; this reduces costs for monitoring. We also showed that gloves swab sampling (e.g., using a cotton swab to recover spores from worker gloves) was more sensitive and cost effective than visual disease scouting; this swabbing technique was rapidly adopted by growers participating in sampling. Our work on fungicide cross-resistance provided data-driven information to growers on how to best adjust their chemical choices to avoid crop loss related to resistance selection in E. necator. The new E. necator genome assembly enabled us to identify the exact genomic location of fungicide target-site genes, create a basis for analyzing genomic structural variations associated with fungicide resistance, and produce a list of candidate genes involved in NTSR. Obj 4: Predicting Resistance Spread Activities: Over 2,500 powdery mildew samples were collected across six major grape growing states. Samples were phenotyped for QoI resistance using qPCR, and sequenced using a pooledseq approach, the first of its kind in an obligate, plant pathogen. Allelic diversity analyses are ongoing to better understand the population dynamics of resistance development. To monitor how spores (particles) spread, we improved our numerical spore dispersion model using graphics processing unit technology, developed a new model for the average wind flow in vineyards, created a new turbulence model for vineyard flows, used our new vineyard models to understand the effectiveness of impaction traps, created an initial model to identify the source of trapped spores given trap data, and built a powdery mildew spread model that we used to examine the effectiveness of different disease scouting strategies. Finally, using publicly available records (CA Pesticide Use Reporting), as well as records solicited from growers across the USA, we were able to capture fungicide use patterns in vineyards from 2012-2018. We were able to capture average number of applications per season, common spray intervals and propensity for tank mixing versus application of a single product. We were also able to see what fungicide groups were most commonly used in US winegrape vineyards. Impacts: Our dispersion model is fast and captures 3D flow features of a vineyard canopy, including the channeling of winds down the rows and directional mixing of spores by turbulence. These models can be used to evaluate best placement of stationary pathogen monitoring tools, given site-specific landscape and environmental conditions. Our fungicide survey provided a clear understanding of fungicide use patterns in the USA. We noted that the use of FRAC 11 fungicides from 2017 until 2020 has declined, which overlaps with the same time period this project has been active in educating the grape pest management communities on FRAC 11 fungicide resistance in grape powdery mildew. There was also an increase in grower's use of contact and biological fungicides, both individual and as tank-mixes, reflective of their learning about the value of these products in fungicide stewardship.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Breeden, S., W. Sanders, C. Hawkins, P. Brannen, C. Johnston, and R. Covington. 2023. Efficacy of Microthiol Disperss sulfur and various surfactants for control of powdery mildew of grape, 2022. Plant Disease Management Reports 17:PF016.
  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Breeden, S., W. Sanders, C. Hawkins, P. Brannen, C. Johnston, and R. Covington. 2023. Efficacy of DMI (FRAC 3) fungicides for control of powdery mildew in a DMI-tolerant Erysiphe necator population, 2022. Plant Disease Management Reports 17:PF017.
  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Pardini, C.A., and A. Esp�nola-Arredondo2023. Facing Fungicide Resistance in Grape Production: A Game Theoretic Approach. Canadian Journal of Agricultural Economics DOI: 10.1111/cjag.12334
  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Stergiopoulos, I., N. Aoun, Q. van Huynh, T. Neill, S. Lowder, C. Newbold, M.L. Cooper, S. Ding, M.M. Moyer, T.D. Miles, C.L. Oliver, J.R. Urbez-Torres, and W. Mahaffee. 2023. Identification of Putative SDHI Target-Site Mutations in the SDHB, SDHC, and SDHD Subunits of the Grape Powdery Mildew Pathogen Erysiphe necator.Plant Disease106: 2310-2320. DOI: 10.1094/PDIS-09-21-1993-RE
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2023 Citation: Lowder, S.R., T. M. Neill, A.B. Peetz, T.M. Miles, M.M. Moyer, C. Oliver, I. Stergiopoulos, S. Ding, and W. Mahaffee.2023. A Rapid Glove-Based Inoculum Sampling Technique to Monitor Erysiphe necator in Commercial Vineyards. Plant Disease (First Look). DOI: 10.1094/PDIS-02-23-0216-RE
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2023 Citation: Oliver, C., M. Cooper, M. Lewis Ivey, P. Brannen, T. Miles, W. Mahaffee, and M.M. Moyer. 2023.Fungicide Use Patterns in Select United States Wine Grape Production Regions. Plant Disease (First Look).DOI: 10.1094/PDIS-04-23-0798-RE
  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Zaccaron, A.Z., Neill, T., Corcoran, J., Mahaffee, W.F., Stergiopoulos, I.2023A chromosome-scale genome assembly of the grape powdery mildew pathogen Erysiphe necator reveals its genomic architecture and previously unknown features of its biology. mbioJun 21:e00645DOI: 10.1128/mbio.00645-23
  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Lowder, S.R., Moyer, M.M., Pscheidt, J., Cooper, M.L., Mahaffee, W.F. (2023 submitted, in review).Grower perspectives on collective action for pest and disease management in vineyards in the western US. Phytofrontiers
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2023 Citation: Sharma, M., T. Neill, H-C.Yang, C. Oliver, W. Mahaffee, R. Naegele, M.M. Moyer, and T.D. Miles.2023. Development of a Competitive PNA-LNA-LAMP Assay to Detect a SNP Associated with QoI Resistance in Erysiphe necator. Plant Disease (First Look). DOI: 10.1094/PDIS-09-22-2027-RE
  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Lowder, S.R., Moyer, M.M., Cooper, M.L., Pscheidt, J., Mahaffee, W.F. (2023 submitted, in review).Information transfer in vineyards: Different grower perspectives, social networks, and how they use information resources for pest and disease management. Phytofrontiers
  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Garcia-Figuera, S, Lowder, S.R., Lubell, M.N., Mahaffee, W.F. McRoberts, N., and Gent, D.H.(2023 submitted, in review).A Social-Ecological Systems Approach to Collective Action in Plant Health. Annual Review of Phytopathology
  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Yang, H.C., Rodriguez J., Hale L. Naegele, R.P.(2023 submitted, accepted minor revisions). Grapevine leaf epiphytic fungal and bacterial communitieis are influenced more by spatial and temporal factors than powdery mildew spray programs. Phytofrontiers.
  • Type: Websites Status: Accepted Year Published: 2023 Citation: FRAME Networks Project Website: https://framenetworks.wsu.edu/
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Wong, A and Mahaffee, WF 2023. A high throughput fungicide tolerance assay for grape powdery mildew,�Erysiphe necator.20th International Reinhardsbrunn Symposium Modern Fungicides and Antifungal Compounds. Friedrichroda, Germany.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Lowder, S.R., Moyer, M.M., Pscheidt, J., Cooper, M.L., Mahaffee, W.F 2022. It takes a village: Understanding barriers to regional cooperatives for pest and disease management. Northwest Center for Small Fruits Research Conference. Corvallis, OR, USA.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Brannen P. 2023. Critical disease updates for Georgia wine producers. Georgia Wine Producers Conference. Ellijay, GA.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Mahaffee, W. 2023. Mission Impossible: Managing Grape Powdery Mildew and Fungicide Resistance. Oregon Vineyard Supply Grower Meeting. McMinnville, OR.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Mahaffee, W. 2023. The Challenge of Managing Grape Powdery: We just cant see it coming. Oregon Wine Symposium. Portland, OR.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Mahaffee, W. 2023. Mission Impossible? Inoculum monitoring as a decision aid. Department of Plant, Soil, and Microbial Sciences, Michigan State University. Lansing, MI.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Mahaffee, W. 2023. Inoculum monitoring as a decision aid. Biopreparedness�Research Virtual Environment (BRaVE) workshop. Department of Energy - Office of Science. Virtual.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Mahaffee, W. 2023. Mission Impossible? Inoculum monitoring as a decision aid. National Predictive Modeling Tool Initiative (NPMTI). Beltsville, MD.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Mahaffee, W. 2023. Fits like a Glove: Novel Approaches to Disease Assessment & Monitoring Fungicide Resistant Powdery Mildew. Sustainable Ag Expo. San Luis Obisbo, CA.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Mahaffee, W. 2022. Finding Needles in Haystacks: Disease Monitoring and Risk Assessment. National Academy of Sciences Workshop on AMRI fungi. Virtual.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Lowder, S.R 2022. It takes a village: Understanding barriers to regional cooperatives for pest and disease management. Northwest Center for Small Fruit Research. Kennewick, WA.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Moyer, M. 2023. Grape Powdery Mildew: As Predictable as the Weather!. Growers Supply Annual Horticulture Show (British Columbia). Virtual.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Moyer, M. 2023. A Pre-Season Checklist for Powdery Mildew Management. LIVE Annual Conference. Chehalem, OR.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Moyer, M. 2022. Best Application Practices in Fruit Production. Great Lakes Expo. Grand Rapids, MI.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: Moyer, M. 2023. Cool vs. Warm Years: Disease and Pest Management. WineVit2023. Kennewick, WA.
  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Margairaz, F., Signh, B., Gibbs, J., Atwood, L., Pardyjak, E., Stoll, R.(2023 re-vised after major revisions, in review).QES-Plume v1.0: A Lagrangian dispersion modelEGUspere(discussion paper)DOI: 10.5194/egusphere-2022-1256
  • Type: Journal Articles Status: Under Review Year Published: 2023 Citation: Ulmer, L, Margairaz,F, Mahaffee, WF, Stoll. R. (2023 submitted).A fast-response model of turbulence and passive scalar transport in row-organized canopiesAgricultural and Foreset Meteorology
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Ulmer, L. 2023. A Fast-Response Modeling System for Particle Dispersion in Vineyards. 103rd AMS Annual Meeting. Denver, CO.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Margairaz, F. 2023. Qes-Plume: A Fast Lagrangian Dispersion Model for Complex Environment. 103rd AMS Annual Meeting. Denver, CO.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Bozorgmehr, B. 2023. Cut-Cell Method: Higher-Order Geometry Representation in Qes-Winds to Calculate More Accurate Wind Fields in Urban Areas and over Complex Terrain. 103rd AMS Annual Meeting. Denver, CO.


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

Outputs
Target Audience:We have numerous target audiences, based on who would benefit most from our various project efforts. Collectively, grape growers, viticulturists, crop consultants, plant disease diagnosticians, and other scientists have accessed and used a multitude of our FRAME project products, both for the improvement of grape production, more efficacious application of disease control strategies, improved fungicide stewardship, increased efficiency in diagnosing fungicide resistance, and advancement of knowledge to direct future research efforts. Grape growers, viticulturists and crop consultants have participated in our formal seminars, regional group meetings, have accessed our newsletters and trade publications. We have also had many personal interactions with industry members regarding fungicide resistance. Peer-networking by growers across growing regions has supported dissemination of project results and increased uptake of recommended mitigation practices. This includes targeted grape growers in Michigan, Indiana, New York, California, Ohio, Georgia, Washington, and Oregon, and indirectly, grape growers across the United States (through trade articles and website access). Crop consultants, diagnosticians, and other scientists have utilized rapid sampling techniques and diagnostic assays to monitor for the presence of grape powdery mildew and resistance QoI and DMI fungicides in their vineyards. Scientists and diagnosticians have used the new screening and ddPCR assays for assessing fungicide resistance phenotypes. Scientists have access to our E. necator genome sequence (published in a peer-reviewed article) assist in their work on understanding E. necator biology and mechanistic host interactions. Improved rapid sampling protocols and kits have been made available to growers and consultants with instructions on return for processing. Our data can also now be used as a reference point for routine monitoring programs and the development of diagnostic assays for fungicide sensitivity. Our recent publications have also been made open-access, so the scientific community can use this information for their own projects. Undergraduate and graduate students have benefited by working directly on projects, participating in writing of manuscripts, and presenting at both virtual and in-person scientific conferences. These activities provide valuable training opportunities in data collection, analysis, and public presentation. Information generated through FRAME has also been incorporated into undergraduate and graduate course lectures. Changes/Problems:No major problems to report (other than additional travel and lab restrictions related to the COVID pandemic). Where possible, when restricted, the team looked for alternative means to complete efforts (i.e., virtual workshops, use of newer, cost-efficient genetic techniques that have been developed in the last few years, etc). We have asked for a 1 year NCE, primarily for Objective 3 and 4 activities, which were more heavily impacted by the pandemic due to their requirements for laboratory access. What opportunities for training and professional development has the project provided?We have used regional group meetings for information sharing, as they are opportunities for peer learning. Information is exchanged among members to encourage adoption of inoculum monitoring and resistance mitigation practices. Stakeholders accessing fungicide resistance data via the FRAME networks Dashboard may evaluate and respond to risk of fungicide resistance in their growing region. Growers participating in FRAME workshops have learned how to both build, and assess, a grape disease program for principles related to pest management and fungicide stewardship. We have also offered numerous training opportunities to undergraduate, graduate, and postdoc mentees in research and scientific communication, with opportunities to work on data collection, analysis, presenting results in written and oral formats. This is noted by the number of publications, posters, and presentations with mentees as first or presenting author. How have the results been disseminated to communities of interest?We have delivered 26 presentations and 8 posters at scientific and industry conferences, delivered 5 FRAME fungicide workshops (3 in-person, 2 virtual) and 3 FRAME-associated workshops, and wrote 2 popular press articles and 8 peer reviewed articles (with 2 more in-review). When growers or consultants submit samples to one or more of our various participating laboratories for fungicide resistance genetic testing, they received customized letters that include their sample results, along with resistance mitigation action plans. Information has also been presented at informal grower networking groups, and through our online interactive FRAC 11 Resistance Dashboard. We also routinely post content to our project website at: framenetworks.wsu.edu, as well as our social media accounts (Twitter - @FRAMEnetworks, Facebook- https://www.facebook.com/FRAMEnetworks). What do you plan to do during the next reporting period to accomplish the goals?The next period is a 5th year, no-cost extension year, where primary activities will be focused on Objective 3 and 4, as expenditures and activities in Objectives 1 and 2 were not impact by COVID and have been predominately spent as originally planned. The exception is some remaining travel funds, which will likely be used to offer additional grower FRAME workshops, based on regional interests and needs. For Objective 3, we will focus on the following: 1) Phenotyping of fungicide-resistant isolates; 2) Sequencing and comparing the transcriptome of six isolates of E. necator when exposed to two fungicides; 3) Studying genomic structural variations in E. necator and their potential impact to fungicide resistance, by sequencing at near-chromosome level the genomes of two more isolates of E. necator and shot-gun sequencing the genomes of an additional 20 isolates; 4) Investigating the impact on SDHI resistance of the mutations that were identified in the sdh subunits of the fungus; 5) Determining the contribution to fungicide resistance in E. necator of a carboxylesterase enzyme, three putative efflux transporters, and a 7-dehydrocholesterol reductase (DHCR7) enzyme that were identified from our genomics work as possible mediators of SDHI or DMI resistance in this pathogen; and 6) Continue processing of grower samples to extend this service for 1 partial growing season (as activities in the NCE need to be completed by mid-June 2023). For Objective 4, we will focus on: 1) Completing our work to map risk of spread from sources within vineyards at the entire vineyard to regional scale. This will consist of linking the new vineyard specific dispersion model (QES-Plume) to the mesoscale weather model WRF to predict spread and thus risk; and 2) Screening and pooling DNA samples for sequencing and data analysis for population genetic studies and writing the resulting 2-3 associated manuscripts.

Impacts
What was accomplished under these goals? Project Management: The fourth annual project meeting was held in-person from 4-7 April 2022 in Hood River, Oregon. Participants included all project PIs and Key Personnel, most of the Stakeholder Advisory group, as well as participating technicians and graduate students. This meeting is where the stakeholder advisory group provides a review and direction on project activities. Obj. 1: Extension and Outreach Obj. 1 - Act. 1: Pre- and post-project surveys. Completed in previous reporting period. Obj. 1 - Act. 2: In-the-field demonstrations of management recommendations. Updates - In Oregon, demonstrated that the reintegration of QoI fungicides into commercial production systems where QoI resistance was at or below detection limits early in the season is feasible without loss of disease management. In Ohio, developed a field trial to tested for QoI resistant E. necator in plots sprayed using intelligent sprayer technology. No differences in incidence was observed. In Georgia, four field trials have been initiated (two in 2021 [completed] and two in 2022 [in process]) to review management methods for control of powdery mildew in the presence of QoI and DMI E. necator resistant populations. Obj. 1 - Act. 3: Develop a "University Service Center" business plan for fungicide resistance testing. Completed in previous reporting period. Obj. 1 - Act. 4: Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups. Updates - Continued to deliver FRAME-specific workshops, both in person and in virtual formats, reaching grape growers across the country. Updated FRAC 11 Fungicide Resistance Dashboard. Stakeholders access data to view spatiotemporal trends in incidence of fungicide resistance. Educational opportunities included a workshop, seminar and neighborhood group meeting. Discussions included challenges and opportunities for adoption of best practices. Regional communication and collaboration were encouraged.. Obj. 2: Group Decisions Obj. 2 - Act. 1: Conduct interviews to identify the externalities negotiating parties are experiencing. Completed in previous reporting period. Obj. 2 - Act. 2: Use game theory to develop compensating mechanisms that induce cooperation. . Updates - Wrote "Fungicide Resistance and Misinformation: A Game Theoretic Approach" under revision in the Can. J. Ag. Econ. Currently developing a model that describes the decision-making process when grape growers can choose between different fungicide use practices (to solve the fungicide resistance problem). Obj. 3: Detection and Monitoring Sub-Obj. 3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Act. 1: Development of PCR based diagnostic assays. Updates - PCR diagnostics have been available now for 2 years. Processed stakeholder samples (> 2,000 across the country). Obj. 3.1- Act. 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Updates - A draft of a paper for peer-reviewed publication is being prepared for submission in Winter 2022. Obj. 3.1 - Act. 3: Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance. Updates - A 3rd year of data demonstrated that a variety of sampling strategies are effective at sampling FRAC 11 resistance but ToughSpots tend to be more effective on DMI and SDHI resistance. Identified significant cross resistance in Botrytis cinerea for many of the powdery mildew fungicides during 2018-2021. This is an issue in FRAC codes 11, and 7 which have effective on powdery mildew and Botrytis. Sub-Obj. 3.2 - Monitoring for Fungicide Resistance. Obj. 3.2 - Act. 1: Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance. Updates - Spore capture devices (CA, OR) and glove-swabs (GA, MI, OR, WA) were used to monitor inoculum occurrence and incidence of FRAC 11 resistance. A 3nd year of data demonstrated that gloves swab sampling was more sensitive and cost effective than visual disease scouting and as effective spore samplers for monitoring the presence of grape powdery mildew at a significantly lower cost than both. Obj. 3.2 - Act. 2: Phenotype for fungicide sensitivity. Updates - Continued to bioassay individual isolates to confirm molecular resting and identify new resistant phenotypes. Expanded the profile of tested materials to include multiple chemistries in the FRAC 3, 7, 9, 11, 13, 50 and U06 groups. Sub-Obj. 3.3 - Understanding the Basis of Resistance. Obj. 3.3 - Act. 2: Create reference genome of E. necator for comparative genomic and transcriptomic approaches. Updates -. Additional gene annotation refinements were carried out in order to increase their accuracy. To elucidate the role of genomic structural variations in fungicide resistance in E. necator, we attempted to generate chromosome-level assemblies of three more isolates by long-read sequencing. High-molecular weight DNA was isolated from the isolates and was submitted for genome sequencing using HiFi, the latest sequencing technology of Pacific Biosciences. The genomes of 5 isolates were sequenced by illumina and are currently analyzed. Obj. 3.3 - Act. 3: Identification of genomic markers associated with fungicide resistance. Updates - An in silico reconstruction of the ergosterol biosynthesis pathway in E. necator was performed since this is the target of DMI fungicides. Most genes in the pathway were conserved, except of Erg4 and Erg5 that catalyze the last two steps in the biosynthesis of ergosterol in fungi. Their absence explains why E. necator does not produce ergosterol. We propose that 7-dehydrocholesterol reductase (DHCR7), a , is involved in the synthesis of 24-methylenecholesterol, the main sterol in the membranes of E. necator. The genome of E. necator was further exploited to produce a targeted list of candidate genes involved in fungicide resistance. A total of 46 genes were predicted as candidates involved in fungicide resistance by efflux transport or enzymatic detoxification. Functional analysis of three of these candidate genes encoding ABC transporters is currently underway, by means heterologous expression in a drug hypersensitive strain of yeast. Obj. 4: Predicting Resistance Spread Sub-Obj. 4.1: Conduct seasonal monitoring of isolates for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Act. 1: Sample collections and phenotyping. Updates - More than 2000 samples collected from 2018-2020 have been organized and pooled into 230 samples for sequencing. Agilent probes have been acquired and protocols are currently being tested. Amplicon markers were developed using the E. necator genome developed in Obj. 3, and have been tested against powdery mildew from table grape vineyards in California. Obj. 4.1 - Act. 2: Genome sequencing and allelic diversity analyses. Updates - Samples are being prepared for sequencing submission. Sub-Obj. 4.2 - Dispersion Tracking and Population Prediction. Obj. 4.2 - Act. 1.1: Mapping resistant isolate spread risk. Updates - Development, finalization, and testing of numerical models that faithfully describe the transport of spores in grape vineyards. This includes continued development and submission of a publication that describes QES-Winds' grape vineyard wind-flow model and continued development of QES-Plume's particle transport model. Obj. 4.2 - Act. 1.2: System validation and evaluation. Updates - Used our models to describe the footprint that a spore sampler 'sees' and developed a powdery mildew epidemiological model that we could use to understand the effectiveness of different disease scouting methods. Sub-Obj. 4.3 - Fungicide Record Evaluation. Updates - Record collection is complete. Finished summarizing / data clean up, and have begun writing the paper summarizing collected fungicide survey data.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2022 Citation: Stergiopoulos, I., Aoun, N., van Huynh, Q., Neill,T., Lowder, S., Newbold, C., Cooper, M.L., Ding, S., Moyer, M.M., Miles, T.D., Oliver, C.L., �rbez-Torres, J.R., and W. Mahaffee. 2022. Identification of SDHI target-site mutations in the SDHB, SDHC, and SDHD subunits of the grape powdery mildew pathogen Erysiphe necator. Plant DiseaseFirst Look. DOI: 10.1094/PDIS-09-21-1993-RE
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Margairaz, F., H. Eshagh, A.N. Hayati, E.R. Pardyjak and R. Stoll. 2022.Development and evaluation of an isolated-tree flow model for neutral-stability conditions.Urban Climate 42:101083.DOI: 10.1016/j.uclim.2022.101083
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Torkelson, G., T. Price, and R. Stoll. 2022.Momentum and Turbulent Transport in Sparse, Organized Vegetative Canopies.Boundary-Layer Meteorology. DOI: 10.1007/s10546-012-9796-4
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zaccaron, A.Z., and I. Stergiopoulos. 2021.Characterization of the mitochondrial genomes of three powdery mildew pathogens reveals remarkable variation in size and nucleotide composition. Microbial Genomics. DOI: 10.1099/mgen.0.000720
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Pardini, C.A., A. Esp�nola-Arredondo, and M.M. Moyer. 2022. Cooperation and Compensation to Mitigate Fungicide Resistance. Am. J. Enol. Vitic 73: 190-195. DOI: 10.5344/ajev.2022.21052
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2022 Citation: Mahaffee, WF, Margairaz,F, Ulmer,L, Bailey, BN, Stoll. R. 2022. Feature article: Catching spores: linking epidemiology, pathogen, biology and physics to ground-based inoculum monitoring.Plant Disease First Look. https://doi.org/10.1094/PDIS-11-21-2570-FE
  • Type: Journal Articles Status: Under Review Year Published: 2022 Citation: Pardini, C.A., and A. Esp�nola-Arredondo.(submitted). Facing Fungicide Resistance in Grape Production: A Game Theoretic Approach. Canadian Journal of Agricultural Economics.
  • Type: Journal Articles Status: Accepted Year Published: 2022 Citation: B. Warres, S. Breeden, W. Sanders, T. Rios, A. Cheh, P. Brannen, D. Rogers and R. Covington. 2022. Assessment of interactions between sulfur and DMI fungicides to control powdery mildew in the presence of a DMI-resistant Erysiphe necator population in Georgia, 2021. Plant Disease Management Reports16:PF010.
  • Type: Journal Articles Status: Accepted Year Published: 2022 Citation: B. Warres, S. Breeden, W. Sanders, T. Rios, A. Cheh, P. Brannen, D. Rogers and R. Covington. 2022. Efficacy of DMI (FRAC 3) fungicides for control of powdery mildew in a DMI-tolerant Erysiphe necator population, 2021. Plant Disease Management Reports 16:PF009
  • Type: Journal Articles Status: Under Review Year Published: 2022 Citation: Ulmer, L, Margairaz,F, Bailey, BN, Mahaffee, WF, Pardyjak, E, Stoll. R. (submitted). A fast-response, wind angle-sensitive model for predicting mean winds in row-organized canopies.Agricultural and Foreset Meteorology.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Zaccaron A. Z, Jorge De Souza J. and Stergiopoulos I. 2022. Assembly and annotation of the mitochondrial genomes of four powdery mildew pathogens reveals remarkable variation in size and nucleotide composition. 31st Fungal Genetics Conference. Pacific Grove, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Samaras A., Zaccaron A. Z., and Stergiopoulos I. 2022. The role of ABC transporters in resistance to SDHI fungicides in the obligate fungal pathogen Erysiphe necator. 31st Fungal Genetics Conference. Pacific Grove, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Heger, Lexi et al. 2022. Advances in molecular and optical detection strategies for grape downy mildew. American Phytopathological Society. Pittsburg, PA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Thompson, S. et al. 2022. Bridging the gap between powdery mildew genomics and valuable culturing methods of Erysiphe necator and Podosphaera aphanis. American Phytopathological Society. Pittsburg, PA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Sharma, N. et al. 2022. Assessment of QoI and CAA fungicide resistance of Plasmopara viticola populations in vineyards of the Great Lakes region in the United States of America. American Phytopathological Society. Pittsburg, PA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Lewis Ivey, M.L. 2022. Precision Application Technologies and Stakeholder Communication. National Academies of Sciences, Engineering and Medicine - Forum on Microbial Threats. Washington, DC.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Espinola-Arredondo, A. 2022. Fungicide Resistance and Misinformation: A Game Theoretic Approach. 27th Annual Conference of the European Association of Environmental and Resource Economists. Virtual.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Lowder, S.R 2022. Rapid Sampling Technique to Monitor Erysiphe necator More Effective Than Visual Scouting. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Moyer, M.M 2022. Improving Confidence and Understanding of Fungicide Program Design in USA Grape Growers Through Peer-Interactive Extension Programming. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Heger, L. 2022. Advances in molecular and optical detection strategies for grape downy mildew. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Thompson, S. 2022. Bridging the gap between powdery mildew genomics and valuable culturing methods of Erysiphe necator and Podosphaera aphanis. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Sharma, N., Heger, L., Neugebauer, K., and Miles, T. 2022. Assessment of QoI and CAA fungicide resistance of Plasmopara viticola populations in vineyards of the Great Lakes region in the United States of America. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Oliver, C.L 2022. Influence of Sustainability Programs on Fungicide Stewardship Practices in Pacific Northwest United States Vineyards. 9th International Workshop on Grapevine Downy and Powdery Mildew. Cremona, Italy.


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

Outputs
Target Audience:The target audience includes grape growers, vineyards managers, vineyard interns, vineyard technicians, vineyard laborers, professional crop consultants, Extension professionals, plant health diagnostic clinics, chemical manufacturing companies, chemical distribution companies, industry organizations, undergraduate students, graduate students, and fellow researchers in the fields of fungicide resistance and pathogen dissemination. We have delivered results of the research through formal seminars, regional grower group meetings, Extension newsletters, trade publications and personal interactions with industry members. Peer-networking by growers across growing regions has supported dissemination of project results and increased uptake of recommended mitigation practices including in-field testing. Outreach presentations have been conducted in 7 states across the country. FRAME-specific fungicide spray program full-day virtual workshops (half-day of presentations, half-day of synchronous activities) have been provided in 12 states. Changes/Problems:We have not experienced major changes, except for unprecedented challenges associated with COVID-19 travel and workspace restrictions. This has impacted some members of our team more than others; however, given we are a smaller project, we were able to adopt and shift some of the necessary workload between groups and have been able to keep largely on-time for most project efforts. In some ways, COVID-19 has actually helped teams, such as Extension, due to the forced wide-spread adoption of distance learning, that have allowed us to reach regional grower groups that would normally be difficult to reach due to costs assocaited with time and travel. What opportunities for training and professional development has the project provided?Members of the Extension team have conducted multiple workshops and training sessions across the country since June 2020 (the end of the previous reporting timeline). This includes one-on-one training sessions for growers in multiple states, 4 FRAME-specific fungicide resistance educational virtual workshops, 23 grower presentations, and a poster at a virtual grower meeting. In-person meetings could not be held due to COVID-19 gathering and travel restrictions. There are multiple undergraduate and graduate students who are currently being trained on this project (7 graduate students, and several undergraduate (5) lab rotations). These students have participated in developing newsletter articles, a popular press article, and several peer reviewed papers (2 published, 1 submitted). Graduate students have also presented posters at the American Phytopathological Society in August 2020 and 2021 and given presentations at four separate scientific conferences. There are also 3 postdoctoral research associates who have participated in developing a peer reviewed paper, delivered 3 presentations and 2 posters at grower and scientific meetings. How have the results been disseminated to communities of interest?We have delivered 31 presentations, 4 virtual FRAME fungicide workshops, 1 popular press article, a blog post, 4 peer reviewed articles, and attended 6 scientific conferences. Our team also developed an interactive dashboard of FRAC 11 fungicide resistance results that was updated in real time during the 2021 growing season. Unfortunately, direct interactions and conference participation were limited to virtual methods due to COVID-19. In addition, when growers or consultants submit samples to one or more of our various participating laboratories for fungicide resistance genetic testing, they receive customized letters that include their sample results, along with resistance mitigation action plans. We also routinely post content to our project website at: framenetworks.wsu.edu, as well as our social media accounts (Twitter - @FRAMEnetworks, Facebook -https://www.facebook.com/FRAMEnetworks). What do you plan to do during the next reporting period to accomplish the goals?Objective 1: Extension and Outreach Obj. 1 - Activity 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United States.Plan of action for year 4 - Begin developing post-project survey questions in fall of 2021 for dissemination in 2022. Obj. 1 - Activity 2: In-the-field demonstrations of management recommendations. Plan of action for year 4 - In-field demonstrations will not occur in year 4 due to timing of the project relative to the growing season. Obj. 1 - Activity 3: Develop a "University Service Center" business plan for fungicide resistance testing. Plan of action for year 4 - Complete compilation of protocols and business plans based on the experiences from the 2020-2021 WSU IAREC lab setup with protocols and budgets from two external labs. Obj. 1 - Activity 4:Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Plan of action for year 4 - Will continue to host regional workshops, write additional extension articles, and podcasts. Will update and expand online educational content to address the current challenges of in-person training. Maintain the mapping tool to report current and historical survey results in real-time for all participating growing regions. Expand extension efforts to further reach constituents across the country. Objective 2: Group Decisions Obj. 2 - Activity 1:Conduct interviews to identify the externalities negotiating parties are experiencing (i.e., how their pesticide use affects their neighbors' production).Plan of action for year 4 - We are in the process of completing drafting the paper for this activity for submission in 2022. Obj. 2 - Activity 2: Use game theory to develop compensating mechanisms that induce cooperation and aid fungicide stewardship by inducing cooperative behavior among growers, crop consultants and chemical companies. Plan of action for year 4 - We are currently working on a signaling game that characterizes the lack of information that some growers face about the severity of fungicide resistance. We consider a context in which two grape growers have already decided to cooperate (reduce their fungicide usage), however, one of them has more information about the severity of fungicide resistance. We aim to evaluate their strategic behavior and identify under which cases growers will tend to deviate from cooperation (i.e., increase their fungicide usage) Objective 3: Detection and Monitoring Sub-Objective 3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Activity 1:Development of PCR based diagnostic assays.Plan of action for year 4 - This activity has been completed. Obj. 3.1 - Activity 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Plan of action for year 4 - Finish analysis of results from the separate labs and complete the paper for submission in Winter 2021. Obj. 3.1 - Activity 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Plan of action for year 4 - Continue to monitor the incidence and dynamics of mutations in FRAC 7 - associated genes in powdery mildew from CA, OR, WA, and MI vineyards. Our toxicity assays have shown that next to target-site mutations, there is an additional mechanism in E. necator that mediates tolerance to SDHIs. We will complete comparative transcriptomic analysis to determine these alternative mechanisms. Sub-Objective 3.2 - Monitoring for Fungicide Resistance. Obj. 3.2 - Activity 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Plan of action for year 4 - In CA, commercial vineyards will be monitored for fungicide resistance by surveying powdery mildew colonies and through a network of fixed, rotorod spore traps. In OR, we are monitoring E. necator and fungicide resistance by deploying 25 spore traps in the region. In OR, MI, GA, and WA, vineyards will be intensively sampled using leaf, glove, and isolate sampling to examine which method is the efficient for allowing an accurate grower decision. This sampling will partially be in collaboration with a large consulting group to cover more vineyards on the west coast. Obj. 3.2 - Activity 2:Phenotype for fungicide sensitivity.Plan of action for year 4 - We will repeat the limited introduction of FRAC 3 and 11 products back into spray programs in cooperating vineyards to assess the viability for product use and maintenance of field level control of grape powdery mildew. We will continue to increase the number of live cultures tested for our new expanded fungicide sensitivity profile. Sub-Objective 3.3 - Understanding the Basis of Resistance. Obj. 3.3 - Activity 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides. Plan of action for year 4 - See Obj. 3.3 Activity 3. Obj. 3.3 - Activity 2:Create reference genome ofE. necatorfor comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Plan of action for year 4 - Paper was published. Activity is complete. Obj. 3.3 - Activity 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis. Plan of action for year 4 -We will further in building genomic and transcriptomic resources for E. necator to understand the molecular basis of fungicide resistance in this pathogen. We will also continue to monitor for SDHI target-site mutations in populations of E. necator from different US states. Objective 4: Predicting Resistance Spread Sub-Objective 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Activity 1:Sample collections and phenotyping. Plan of action for year 4 - New postdoctoral researcher and technician will build amplicon libraries for pooled powdery mildew samples and submit them for sequencing. Sequenced samples will be analyzed for genetic variability. Obj. 4.1 - Activity 2: Genome sequencing and allelic diversity analyses. Plan of action for year 4 - Primers and probes will be finalized with the completion of the genome sequences in Spring 2021. Sub-Objective 4.2 - Dispersion Tracking and Population Prediction. Obj. 4.2 - Activity 1:Mapping resistant isolate spread risk. Plan of action for year 4 - Code cleanup has been completed. Currently in the process of drafting papers for peer-reviewed publication. Obj. 4.2 - Activity 2: System validation and evaluation. Plan of action for year 4 - the implementation of the QES dispersion modeling system with a focus on real world application of dispersion with full topography and flow fields linked to weather. We will also continue to work on efficiency of QES to make its interface, execution, and analysis smooth. This will enable to project to move from engaging researchers to engaging practitioners. Sub-Objective 4.3 - Fungicide Record Evaluation. Plan of action for year 4 - We will conclude record collections in Fall 2021 and will begin data analysis. A paper will be drafted for publication in 2022.

Impacts
What was accomplished under these goals? Overall- The third annual project meeting was held virtually 23 - 24 Feb, 2021. Participants included all project PIs and Key Personnel, most of the Stakeholder Advisory group, as well as participating technicians and graduate students. Objective Teams have and continue to host regular web conferences from monthly to quarterly (depending on nature of the objective). Objective 1 - We have learned how growers relate to information about fungicide stewardship. We have developed improved extension curriculum for optimized information uptake and adoption, and have a solid resource set on our website for regular grower interaction. Objective 2 - We have learned what drives grower's willingness to cooperate, and will be able to prescribe policy recommendations providing a different approach to subsidies or penalizations. Objective 3 - We have rapid sampling and detection tools that provide near real-time results for grape growers. Objective 4 - We have learned how powdery mildew can spread in a vineyard, and observed general fungicide application practices. Objective 1: Extension and Outreach Obj. 1 - Act. 1: Pre- and post-project surveys. Accomplishments - Published presurvey results in AJEV. Obj. 1 - Act. 2: In-the-field demonstrations.Accomplishments - Five field trials have been initiated in GA (three in 2020 [completed] and two in 2021 [in process]) to review management methods for control of powdery mildew in the presence of FRAC 3 and 11Erysiphe necator resistant populations. All powdery mildew survey data collected to date (2017-2020) were collated and a Dashboard was designed to display data by year, region and grape category (raisin, table, wine and juice). Obj. 1 - Act. 3: Develop a "University Service Center" business plan.Accomplishments - Consulted with local growers and adjusted the testing facility procedures to decrease the amount of time between sample receipt and result dispersion to growers. Obj. 1 - Act. 4:Develop workshops and other classic extension tools. Accomplishments - Delivered an additional 36 FRAME presentations with a cumulative attendance of approximately 2,400 individuals. The audiences included both the industry and the scientific community, with 23 of the 31 presentations targeted to industry. Our FRAME fungicide spray program workshop was reformatted to an asynchronous virtual format containing a series of 5 videos and a half-day interactive design section. This workshop was presented 4 times with attendees from 12 states (IA, IN, MA, MD, MI, MN, ND, NJ, OH, PA, WA, and WI). Additionally, there have been 5 student/technician posters, 1 blog post, 1 trade article, 4 newsletters, and 8 videos since June 2020. Objective 2: Group Decisions Obj. 2 - Act. 1:Conduct interviews to identify the externalities negotiating parties are experiencing.Accomplishments - We finished collecting data from two surveys: grape growers and crop consultants. The paper based on the grower survey is under revision in Journal of Wine Economics (submitted 05/20/2021) and are developing a research paper focused on the strategic behavior of grape growers when their choices of fungicide levels generate fungicide resistance which is under review in the European Review of Agricultural Economics. Objective 3: Detection and Monitoring Sub-Objective 3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Act.1:Development of PCR based diagnostic assays.Accomplishments - The PCR diagnostics are now in full-use for real-time diagnostics of grower-submitted samples. Obj. 3.1 - Act.2: Developing isothermal diagnostic tools.Accomplishments - A draft of a paper for peer-reviewed publication is being prepared for submission in Winter 2021. We are not pursuing this diagnostic tool further, per the recommendations of our stakeholder advisory group. Obj. 3.1 - Act. 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Accomplishments - Continued expanding the collection of resistant isolates, including isolates resistant to multiple FRAC group fungicides. Sub-Objective 3.2 - Monitoring for Fungicide Resistance. Obj. 3.2 - Act. 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Accomplishments - Spore capture devices (CA, OR) and glove-swabs (GA, MI, OR, WA) were used to monitor inoculum occurrence and incidence of FRAC 11 resistance. Processed more than 5213 swabs samples for FRAC 11 and FRAC 3 genetic markers in OR, and 300 in WA. Obj. 3.2 - Act. 2:Phenotype for fungicide sensitivity.Accomplishments - We collaborated with 3 OR growers that quit using FRAC 3 and 11 fungicides to see if thesefungicides could be rotated back into their program as single applications and still achieve successful field control. We continued to bioassay individual isolates to confirm molecular resting and identify new resistant phenotypes. We expanded the profile of tested materials to include multiple chemistries in the FRAC 3, 7, 9, 11, 13, 50 and U06 groups. Sub-Objective 3.3 - Understanding the Basis of Resistance. Obj. 3.3 - Act. 2:Create reference genome ofE. necator.Accomplishments - Gene annotation of the assembled E. necator genome was completed, with an estimated completeness of 97%. Obj. 3.3 - Act. 3: Identification of genomic markers associated with fungicide resistance.Accomplishments - Functional annotations of major gene categories involved in drug resistance were performed. Further identification marker examinationrevealed that the genotype of EnFRAME01 is similar to east coast genotypes, rather than the other west coastisolates. Transcriptome of EnFRAME01 at six time points during infection has been sequenced and found3,000 genes whose expression varied significantly through time. Among four isolates of E. necator included in the analyses, no allelic variation was observed in cytb apart from the G143A mutation associated with resistance to QoIs. Objective 4: Predicting Resistance Spread Sub-Objective 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Act. 1:Sample collections and phenotyping.Accomplishments - Collected more than 1300 samples representing fields from CA, WA, OR, and GA using gloves and trap plants and tested for QoI resistance. A subset of samples with sufficiently high DNA were selected for sequencing, and several products tested for improving DNA quality prior to sequencing. A postdoctoral researcher was hired and a new technician was hired (both will start September 2021). Obj. 4.1 - Act. 2: Genome sequencing and allelic diversity analyses.Accomplishments - Genome sequence was published and amplicon based sequencing probes (for ~1400 genes) were developed. Sub-Objective 4.2 - Dispersion Tracking and Population Prediction. Obj. 4.2 - Act.1:Mapping resistant isolate spread risk.Accomplishments - The QES GPU based flow solver, the real-time dispersion and spore tracking system, development was completed thoughdynamic coupling. The model was used in initial studies to examine how spore trap placement impacts which vines are sampled in the field. We also developed a powdery mildew 3D disease spread model that incorporates key biophysical components that contribute to spore movement. The model was used in initial studies on the effectiveness of different disease scouting methodologies (e.g., visual identification, impaction traps). Obj. 4.2 - Act. 2: System validation and evaluation.Accomplishments - Models are currently being verified and two publications are being drafted which will be submitted early in 2022. Sub-Objective 4.3 - Fungicide Record Evaluation.Accomplishments - Expanded collected records to 55 CA, 2 GA, 5 MI, 1 OH, 67 OR and 19 WA vineyards for a total of 152 participating vineyards. Additionally, preliminary analyses have begun.

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Warres, B., Breeden, S., Winkles, J., Severns, P., Brannen, P., Rogers, D., Covington, R., Mahaffee, W., and Neill, T. 2020. Fungicide comparisons for powdery mildew management in a fungicide-resistant Erisiphe necator population, 2019. Plant Dis. Mgmt. Rep.14:PF014.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Warres, B., Breeden, S., Winkles, J., Severns, P., Brannen, P., Rogers, D., Covington, R., Mahaffee, W., and Neill, T. 2020. Efficacy of sulfur and myclobutanil combinations for control of grapevine powdery mildew in Georgia, 2019.Plant Dis. Mgmt. Rep.14:PF015.
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Warres, B., Johnson, K., Busher, K., Cameron, C., Brannen, P., Rogers, D., Covington, R., Mahaffee, W., and Neil, T. 2021. Fungicide comparisons for powdery mildew management in a fungicide-resistant Erysiphe necator population, 2020.Plant Dis. Mgmt. Rep.15:PF012
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Warres, B., Johnson, K., Busher, K., Cameron, C., Brannen, P., Rogers, D., Covington, R., Mahaffee, W., and Neil, T. 2021. Assessment of interactions between sulfur and DMI fungicides to control powdery mildew in the presence of a DMI-resistant Erysiphe necator population in Georgia, 2020.Plant Dis. Mgmt. Rep.15:PF013
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Oliver, C., Cooper, M., Lewis-Ivey, M., Brannen, P., Miles, T., Mahaffee, W., and Moyer, M.2021. Assessing the United States Grape Industrys Understanding of Fungicide Resistance Mitigation Practices Am. J. Enol. Vitic.72: 181-193. https://doi.org/10.5344/ajev.2021.20062
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Bozorgmehr, B., Willemsen, P., Gibbs, J., Stoll, R., Kim, J., and Pardyjak, E. 2021. Utilizing dynamic parallelism in CUDA to accelerate a 3D red-black successive over relaxation wind-field solverEnvironmental Modelling & Software: with Environment Data News.137
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Warres, B., Johnson, K., Busher, K., Cameron, C., Brannen, P., Rogers, D., Covington, R., Mahaffee, W., and Neil, T. 2021. Efficacy of DMI (FRAC 3) fungicides for control of powdery mildew in a DMI-tolerant Erysiphe necator population, 2020. Plant Dis. Mgmt. Rep.15:PF014
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Zaccaron, A., De Souza, J., and Stergiopoulos, I. 2021 The mitochondrial genome of the grape powdery mildew pathogen Erysiphe necator is intron rich and exhibits a distinct gene organization. Scientific Reports. 11:13924
  • Type: Other Status: Published Year Published: 2021 Citation: Oliver, C. and Moyer, M. 2021. FRAME Network: Fungicide Resistance in Wine Grapes. WineVit. Virtually. (Poster)
  • Type: Other Status: Published Year Published: 2021 Citation: Newbold, C., and Mahaffee, W. 2021. Phenotypic variability among Erysiphe necator isolates. American Phytopathological Society. Virtually. (Research On Demand)
  • Type: Other Status: Published Year Published: 2021 Citation: Lowder, S., and Mahaffee, W. 2021. Reduction of the G143A mutation in Erysiphe necator overwintering chasmothecia. American Phytopathological Society. Virtually. (Research On Demand)
  • Type: Other Status: Published Year Published: 2021 Citation: Sharma, N., and Miles, T. 2021. Assessment of fungicide resistant Plasmopara viticola populations in vineyards of the Great Lakes region. American Phytopathological Society. Virtually. (Research On Demand)
  • Type: Other Status: Published Year Published: 2021 Citation: Yang, H., and Miles, T. 2021. Grape powdery mildew spray programs affect non-target fungi populations differentially in California vineyards. American Phytopathological Society. Virtually. (Research On Demand)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Mahaffee, W. 2020. Finding Needles in Haystacks: Inoculum Monitoring as a Decision Aid. Western Region IPM Center IPM hour Webinar series. http://youtu.be/ewG-Fv5gS4o.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer, M. 2020. A Casual Conversation on Grape Powdery Mildew. Thirsty Thursday Westover Viticulture Grower Meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Mahaffe, W. 2020. Improved Scouting Methods for Powdery Mildew in Vineyards. Sustainable Ag Expo. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Mahaffe, W. 2020. Rollers, Wakes and Vortices and Other Considerations in Locating Samplers. APS Webinar. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Warres, B., and Brannen, P. 2020. Efficacy of Fungicides for Grape Powdery Mildew Control in the Presence of Fungicide-Resistant Erysiphe necator Populations in Georgia. Cumberland-Shenandoah Fruit Workers Conference. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Brannen P. 2021. Fungicide Resistance Management for Powdery and Downy Mildews. Maryland Grape and Wine Industry Annual Conference. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Mahaffe, W. 2021. Updates on fungicide resistance monitoring. LIVE technical meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Moyer, M. 2021. FRAME-ing Grape Powdery Mildew Management for the West Coast. Syngenta Crop Consultant Summit. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Lowder, S. 2021. Fits like a glove: A new method to monitor grape powdery mildew and FRAC 11 fungicide resistance. Orchard and Vineyard Supply Grower Meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Mahaffe, W. 2021. New approaches for improving disease scouting. Coastal Viticulture Consultants. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Pardini, C. 2020. Facing Fungicide Resistance in Grape Production: A Game Theoretic Approach. AERE at SEA Virtual Meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Warres, B., and Brannen, P. 2021. Fungicide resistance survey of Georgia wine grapes and fungicide efficacy in the presence of a resistant Erysiphe necator population. Georgia Association of Plant Pathologists meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Warres, B., and Brannen, P. 2021. Survey of QoI fungicide resistance in Erysiphe necator populations in Georgia. Southern Fruit Workers meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Brannen, P. 2021. Downy and Powdery mildew resistance management update. Georgia Wine Producers Conference. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Espinola-Arredondo, A. 2021. Fungicide Resistance and Misinformation: A Game Theoretic Approach. Montreal Workshop on Resource and Environmental Economics. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Moyer, M. 2021. Alternative Approaches for Powdery Mildew Management: Fungicide Resistsance Update. Washington Advancements in Viticulture and Enology (WAVE). Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Aoun N. 2021. Monitoring for SDHI resistance in field isolates of Erysiphe necator. Oregon Wine Research Institute Spring Webinar. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Lowder, S. 2021. Fits like a glove: Monitoring powdery mildew and what that can tell us about QoI resistance. Oregon Wine Research Institute Spring Webinar. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Lowder, S., Neill, T., Miles, T., Moyer, M., Ding, S., Oliver, C., Butler, C., Walgenbach, P., and Mahaffee, W. 2021. Spatial and temporal trends in G143A caused QoI resistance in Erysiphe necator across the growing season in the Western US. American Pathological Society - Pacific Division. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Newbold, C., and Mahaffee, W. 2021. Fitness difference in Quinilone outside inhibitor fungicide resistant Erysiphe necator. American Pathological Society - Pacific Division. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Renault, M., Pardyjak, E., and Stoll, R. 2021. Fast-Response, High-Resolution Flow Solver for Vegetation Canopies. 34th Conference on Agricultural and Forest Meteorology/5th Conference on Biogeosciences. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Ulmer, L., Miller, M., Bailey, B., Mahaffee, W., Pardyjak, E., and Stoll., R. 2021. A Semi-Empirical Wind-Resolving Model for Predicting Transport Processes in Row-Organized Agricultural Canopies. 34th Conference on Agricultural and Forest Meteorology/5th Conference on Biogeosciences. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Lowder, S. 2021. Early Powdery Mildew Detection and Fungicide Resistance. Orchard and Vineyard Supply Grower Meeting. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: DeLong, J., Naegele, R., Saito, S., Wang, F., and Xiao, C. 2021. Grape powdery mildew spray programs affect non-target fungi populations differentially in California vineyards. American Phytopathological Society. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Oliver, C. 2021. Using Historical Fungicide Records to Evaluate Use Patterns. American Phytopathological Society. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Mahaffe, W. 2021. Mission Impossible? Inoculum monitoring as a decision aide. University of Maryland. Virtually.
  • Type: Other Status: Published Year Published: 2021 Citation: Lowder, S., and Warres, B. 2021. Early-season scouting for grape powdery mildew. Good Fruit Grower. https://www.goodfruit.com/early-season-scouting-for-grape-powdery-mildew/ (trade article)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Deford, L., Stoll, R., and Pardyjak, E. 2021. Development of a Low-Cost Aspirated Temperature Measurement Radiation Shield. 34th Conference on Agricultural and Forest Meteorology/5th Conference on Biogeosciences. Virtually.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Oliver, C. 2021. Understanding Fungicide Use Patterns in Vineyards through Historical Use Records. American Society for Enology and Viticulture. Virtually.
  • Type: Other Status: Published Year Published: 2021 Citation: Oliver, C.2021. New FRAME networks Resources for Growers. Washington State University Viticulture and Enology Extension News - Spring 2021. http://s3-us-west-2.amazonaws.com/sites.cahnrs.wsu.edu/wp-content/uploads/sites/66/2021/04/15161804/2021-Spring-VEEN-FINAL.pdf (newsletter)
  • Type: Other Status: Published Year Published: 2021 Citation: Lowder, S., and Mahaffee, W. 2021. Hands on information: Updates on rapid sampling techniques for grape powdery mildew. Oregon Wine Research Institute Technical Newsletter - Spring 2021. (newsletter)


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

Outputs
Target Audience:The target audience for this project is broad and includes: grape growers, vineyards managers, vineyard interns, vineyard technicians, vineyard laborers, professional crop consultants, Extension professionals, plant health diagnostic clinics, chemical manufacturing companies, chemical distribution companies, industry organizations, undergraduate students, graduate students, and fellow researchers in the fields of fungicide resistance and pathogen dissemination. We are reaching these audiences through one-on-one recommendations and interactions, newsletters, trade publications, regional industry meetings, and academic conferences. Outreach presentations have been conducted in 10 states across the country. FRAME-specific fungicide spray program full day workshops have been provided in 4 states on both coasts. Rapid powdery mildew sampling kits to screen for fungicide resistance have been made available to growers and consultants, along with kits designed to assist growers in determining spray coverage. Kits for fungicide resistance molecular screening assay validation have been created and disseminated to participating labs in 3 states. The FRAME scientific team has also been regularly presenting through scientific venues, including routine publication of research results, along with presentations and posters at scientific conferences such as the American Phytopathological Society annual meeting. Changes/Problems:We continued to have personnel challenges associated with slow hiring processes through the USDA, but many of these were alleviated by the end of Year 2. However, at that time, challenges related to University hiring (freezing of new hires due to financial challenges brought about by COVID-19), arose. While we continue to make excellent progress despite these hiring challenges, they have impacted our ability to spend in salary and wage allocations. Actual field activities (which is a signficant component of this project) were slowed due to COVID-19, but not stopped, so work efforts have proceeded at a similar-to-normal pace. Project Meetings (both routine and annual) will likely move forward on an exclusively digital basis until a more consistent national return-to-work approach has been acheived. The Extension team will also be re-considering education and outreach efforts, to determine how to best deliver those in durable forms that don't require face-to-face gatherings (where those gatherings are restricted). Many stakeholder winter meetings have already gone to ditigal platforms, which will likely provide a positive opporutnity to "interact" with more groups than was possible when travel was required. What opportunities for training and professional development has the project provided?Members of the Extension team have conducted multiple workshops and training sessions across the country since June 2019 (the end of the previous reporting timeline). This includes 2 GA extension agent trainings, one-on-one training sessions for growers in multiple states, 3 FRAME-Specific fungicide resistance educational workshops, 41 grower presentations, and multiple posters at grower meetings. We have also hosted Round-table discussion meetings in California, 3 wine grape spray workshops, and multiple open houses and field days in late summer / early fall of 2019. There are multiple undergraduate and graduate students who are currently being trained on this project (6 graduate students, and several undergraduate lab rotations). These students have participated in developing newsletter articles and are also currently working on a popular press article. They have also given departmental seminars (2), as well as participated in the FRAME resistance training workshops as either participants or working directly with growers on the activities. Graduate students have also presented posters at the American Phytopathological Society in August 2019 and August 2020. How have the results been disseminated to communities of interest?We have delivered 50 presentations, 4 FRAME fungicide workshops, 2 original videos (one on fungicide resistance, one on spore trap assembly), blog posts, email blasts, and direct interactions. In addition, when growers or consultants submit samples to one or more of our various participating laboratories for fungicide resistance genetic testing, they receive customized letters that include their sample results, along with resistance mitigation action plans. We also routinely post content to our project website at: framenetworks.wsu.edu, as well as our social media accounts (Twitter - @FRAMEnetworks, Facebook - https://www.facebook.com/FRAMEnetworks). What do you plan to do during the next reporting period to accomplish the goals?Objective 1: Extension and Outreach Obj. 1 - Act. 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United States.Plan- Submit pre-project survey for publication; begin developing post-project survey questions. Obj. 1 - Act. 2: In-the-field demonstrations of management recommendations. Plan- Consider alternative means of field demonstrations. Obj. 1 - Act. 3: Develop a "University Service Center" business plan for fungicide resistance testing. Plan- Begin compilation of protocols and business plans based on the experiences from the 2020 WSU IAREC lab setup with protocols and budgets from two external labs. Obj. 1 - Act. 4:Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Plan- Continue regional workshops (digitally), write extension articles, and podcasts. Develop accessible mapping tool to report current and historical survey results for all participating growing regions. Objective 2: Group Decisions Obj. 2 - Act. 1:Conduct interviews to identify the externalities negotiating parties are experiencing.Plan- All surveys have been distributed. Our objectives for the Fall 2020 are: (1) analyze data, (2) literature review and (3) prepare a paper. Obj. 2 - Act. 2: Use game theory to develop compensating mechanisms that induce cooperation and aid fungicide stewardship by inducing cooperative behavior among growers, crop consultants and chemical companies. Plan- Currently working on signaling game that characterizes the lack of information growers face about the severity of fungicide resistance. We consider a context in which two grapes growers have already decided to cooperate, however, one of them has more information. We aim to evaluate their strategic behavior and identify under which cases growers will deviate from cooperation. Objective 3: Detection and Monitoring Sub-Obj.3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Act. 1:Development of PCR based diagnostic assays.Plan- Complete FRAC 11 detection manuscript and submit for peer reviewed publication. Continue to optimize isothermal assays for sensitivity and specificity and conduct technology transfer experiments with other partner laboratories. Obj. 3.1 - Act. 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Plan- Finish analysis of results from the separate labs and complete the paper for submission in Spring 2021. Obj. 3.1 - Activity 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Plan- Continue monitoring incidence and dynamics of mutations in the FRAC 7 - genes in powdery mildew from CA, OR, WA, and MI vineyards. Determine the effect of mutations tolerance in E. necator. Develop diagnostic assays for some of the mutations. We will continue to perform target-site gene sequencing as this is currently more cost effective and reduces the response time needed for obtaining accurate results. A multiplex PCR assay is being developed to allow us to amplify 3 SDH genes in a single PCR reaction. Isolates will be collected from vineyards in which the mutations were identified and tested for tolerance to SDHIs using leaf-disc toxicity assays. For mutations that increase the tolerance of the fungus against SDHIs, TaqMan and other PCR-based diagnostic assays will be developed and tested for specificity and sensitivity. High resolution melting analysis will be pursued to simultaneously detect multiple mutations. Sub-Obj. 3.2 - Monitoring for Fungicide Resistance. Obj. 3.2 - Act. 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Plan- In CA, vineyards will be monitored for fungicide resistance by surveying powdery mildew. In OR, monitoring of E. necator and fungicide resistance will be done by deploying 25 spore traps. In OR, MI, GA, and WA, vineyards will be sampled using leaf, glove, and isolate sampling for method validation. Sampling will collaborate with a large consulting group to cover more vineyards on the west coast. Obj. 3.2 - Act. 2:Phenotype for fungicide sensitivity.Plan- We will collaborate with growers that quit using FRAC 3 and 11 fungicides in 2016 no longer have detectable FRAC 11 resistance populations to see if these fungicides can be re-introduced. We continue to bioassay individual isolates to confirm molecular testing and identify new resistant phenotypes. We will optimize rapid testing technique and increase the number of live cultures tested for fungicide sensitivity. Sub-Obj. 3.3 - Understanding the Basis of Resistance. Obj. 3.3 - Act. 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides. Plan- See Obj. 3.3 Act. 3. Obj. 3.3 - Act. 2:Create reference genome ofE. necatorfor comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Plan- Annotations of E. necator genome will be completed utilizing PacBio's Isoform sequencing method to capture full-length transcripts. Using the methods developed during 2020, 5-6 more isolates will be sequenced to provide additional genetic references to capture the variation between sensitive and resistant isolates of E. necator. Obj. 3.3 - Act. 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis. Plan- The new genome assembly of E. necator will be used to search for sources of SDHI resistance. This will be primarily done through (i) identification of differentially expressed genes (DEGs) resulting from exposure to different SDHIs compounds and doses, and (ii) characterization of genomic structural variants (SVs). RNAseq reads will be mapped to our reference E. necator genome assembly with HISAT2 and DEGs will be identified with DESeq2. To identify SVs, 3 to 5 additional E. necator strains showing different patterns of SDHI resistance will be sequenced and assembled at near chromosome level. Sequencing will be mainly performed at 100x depth coverage on the PacBio Sequel II system. In addition, Illumina data will be generated to correct single base errors and small INDELs in the assemblies. The genomes assembled with Canu and whole-genome alignments with NUCmer. Isolates created will be saved for use 1) in future fitness costs experiments, 2) validation of molecular, and 3) as a correlative tool to see if resistant isolates belong to specific sub populations of E. necator. Objective 4: Predicting Resistance Spread Sub-Obj. 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Act. 1:Sample collections and phenotyping. Plan- Collection of 2020 field samples from glove swabs and trap plants will be completed. Pooled isolates will be barcoded and submitted for sequencing. Obj. 4.1 - Act. 2: Genome sequencing and allelic diversity analyses. Plan- Primers and probes will be finalized with the completion of the genome sequences in Spring 2021. Sub-Obj. 4.2 - Dispersion Tracking and Population Prediction. Obj. 4.2 - Act. 1:Mapping resistant isolate spread risk. Plan- Wrap up the code clean-up and final code notation in Winter 2020. Obj. 4.2 - Act. 2: System validation and evaluation. Plan- Will use the validated and accelerated dispersion modeling system in an inverse mode with both idealized and realistic vineyard geometries and distributions of E. necator. Will enable us to understand how data collected by FRAME objectives links to the distribution of E. necator. Sub-Obj. 4.3 - Fungicide Record Evaluation. Plan- Will continue to collect fungicide spray records from participating states.

Impacts
What was accomplished under these goals? Overall- Project meeting held 11 - 12 Mar, 2020, in Napa, CA. Participants included: PIs and Key Personnel, Stakeholder Advisory group, and several technicians and graduate students. Objective Teams host regular (weekly to quarterly) web conferences. Objective 1: Extension and Outreach Obj. 1 - Act. 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United States.Accomplishments - Completed data analysis and drafted paper for publication. Obj. 1 - Act. 2: In-the-field demonstrations of management recommendations. Accomplishments - Due to COVID-19, in-field demonstrations were reduced. A FRAC 3 and 11 fungicide trial was established in GA and sentinel blocks for disease progress and observation were established in OH. In WA, monitoring fungicide trials for FRAC 11 resistance continues. Obj. 1 - Act 3: Develop a "University Service Center" business plan for fungicide resistance testing. Accomplishments - Began collection of budgets, and "dry run" testing facility establishment (purchasing supplies, assay validation, etc.). Obj. 1 - Activity 4:Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Accomplishments - Delivered 50 FRAME presentations since Jun 2019 (Jun-Aug 2019 not included in previous report). Cumulative attendance of 2,800+ individuals. Audiences included: industry and scientific community, with 42 of the 50 presentations targeted to industry. FRAME fungicide spray program workshop was presented in 4 states (WA, OR, CA, and OH). There were 12 student/tech posters, 3 blog posts, 3 trade articles, 6 newsletters, 2 videos, and a podcast since June 2019. Objective 2: Group Decisions Obj. 2 - Act. 1:Conduct interviews to identify the externalities negotiating parties are experiencing.Accomplishments - Finished data collection from 3 surveys. Currently developing a paper that focuses on the survey results. Developed a second paper titled "Facing Fungicide Resistance in Grape Production: A Game Theoretic Approach." This provides insight into behavior of grape growers when their choices generate a negative intertemporal production externality in the form of fungicide resistance. When growers encounter this externality, the noncooperative fungicide level is higher than the socially optimal level. We examine a compensation mechanism to ameliorate fungicide resistance and find that it induces the socially optimal level; however, misinformation about severity of the fungicide resistance generates distortions. Information available to growers about fungicide resistance is essential for its mitigation with the proposed compensation mechanism. If the misinformed grower considers fungicide resistance to be relatively mild, then the misinformed grower has the compensating role. Objective 3: Detection and Monitoring Sub-Obj. 3.1 - Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Act. 1:Development of PCR based diagnostic assays. Accomplishments - A manuscript was published on FRAC 11 molecular diagnostics. Obj. 3.1 - Act. 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Accomplishments - New rapid isothermal tool (LAMP) was developed for FRAC 11 resistance detection. Assay was validated in external labs. During the annual meeting, stakeholders advised that new diagnostics should focus traditional assays. Obj. 3.1 - Act. 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance. Accomplishments - Collected samples from vineyards with FRAC 7 and 13 resistance to establish living lab cultures. Sub-Obj. 3.2 - Monitoring for Fungicide Resistance. Obj. 3.2 - Act. 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance. Accomplishments - Repeated collections in CA, GA, MI, OR, and WA. Demonstrated that gloves swab sampling was more sensitive and cost effective than visual scouting. Also was as effective as spore samplers for monitoring for disease. Processed >4000 swabs samples for FRAC 11 and 3 resistance markers. MI processed ~700 samples from multiple locations. Obj. 3.2 - Act. 2:Phenotype for fungicide sensitivity.Accomplishments - Identified pathogen isolates resistant to FRAC 7 and 13 fungicides. Found >14 isolates with FRAC 7 resistance and 4 with FRAC 13 resistance. Identified 100+ isolates with FRAC 11resistance and 48 with FRAC 3 tolerance. Sub-Objective 3.3 - Understanding the Basis of Resistance. Obj. 3.3 - Act. 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides. Accomplishments - Naturally-occurring isolates were found, so this activity is not needed. Obj. 3.3 - Act. 2:Create reference genome ofE. necatorfor comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms. Accomplishments - Further refined large-scale culturing of E. necator. Produced enough DNA for sequencing of 2 more E. necator isolates. Obtained a chromosome-scale genome assembly for E. necator. Among the assembled scaffolds, 11 are complete chromosomes that contain telomeric repeats at both ends. This new genome is a considerable improvement over the 2014 assembly - now at 7,570 predicted genes with 97% completeness. Developed transcriptomes of 6 E. necator isolates to refine the genome annotation. Obj. 3.3 - Act. 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis. Accomplishments - Transcriptomics studies are underway for determining FRAC 13 resistance. In total this project has collected over 1600 Gbp of RNAseq data. These experiments investigate resistant and sensitive isolates to quinoxyfen (FRAC 13) at various timepoints and conditions, in order to develop a resource for understanding the expression regulation of E. necator. For FRAC 7, we continued USA monitoring and nd identified new SDHI target-site mutations. Target-site mutations were identified in 9 of 12 states, with the majority of strains with such mutations originating from CA and MI. The frequency of FRAC 7 target-site mutations almost doubled in 2019 as compared to 2018, although there are were no reports of reduced field efficacy. Objective 4: Predicting Resistance Spread Sub-Obj. 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Act. 1:Sample collections and phenotyping. Accomplishments - Field collections continued. Probes developed for specific powdery mildew genes to facilitate exome-based pooled sequencing. Obj. 4.1 - Act. 2: Genome sequencing and allelic diversity analyses. Accomplishments - Genome sequencing is still underway. Preliminary identification of primers and probes has begun with the genome annotations provided from Obj. 3. Sub-Obj. 4.2 - Dispersion Tracking and Population Prediction. Obj. 4.2 - Act. 1:Mapping resistant isolate spread risk. Accomplishments - Modernized dispersion modeling numerical code base, enabling the use of graphics processing unit (GPU) technology. This has resulted in a greater than 100x reduction in model execution time for our E. necator spore dispersion model. Obj. 4.2 - Act. 2: System validation and evaluation. Accomplishments - Finalized the physics of vineyard dispersion model by performing a rigorous validation against existing vineyard dispersion data. Journal articles for both advancements will be submitted for publication soon. Sub-Obj 4.3 - Fungicide Record Evaluation. Accomplishments - Collected records from 75 CA, 2 GA, 1 OH, 3 MI, and 13 WA vineyards with a history of QoI resistance testing. Records needed extensive quality control.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Oliver C. 2019. Fungicide Stewardship - Best Practices. FRAME Extension Meeting. Salem, OR
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Wong, A. 2019. Powdery Mildew Biology. FRAME Extension Meeting. Salem, OR
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lowder S. 2019. Rapid sampling techniques to monitor fungicide resistant grape powdery mildew. Mendicino County Grape IPM Conference. Hopland, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Miles T. 2020. Detecting and managing Plant pathogens in Michigan vineyards. Orchard and Vineyard Show. Traverse City, MI
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Warres, B., and Breeden, S. 2020. Wine Grape Powdery and Downy Mildew Resistance Management. Georgia Wine Growers Association. Braselton, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Miles T. 2020. Detecting and managing Plant pathogens in Michigan vineyards. Southwest Michigan Horticultural Days. Benton Harbor, MI
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Mahaffee W. 2020. Recognizing, monitoring, and mitigating fungicide resistance. D&M Chem Inc. Annual Grower Meeting. Moxee, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lowder S. 2020. Powdery Mildew Management: Monitoring Fungicide Resistance. Oregon Wine Symposium. Portland, OR
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Stergiopoulos I. 2020. Fungicide Resistance in Grape Powdery Mildew Across the Western US; What Do We Know So Far. Current Wine and Winegrape Research Course, UC Davis Extension . Davis, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lewis-Ivey, M. 2020. Best practices for utilizing fungicides to prevent fungicide resistance development. Ohio State University, Pesticide Safety Education Program. Akron, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Zaccaron A. 2020. A chromosome-level assembly of the grape powdery mildew fungus Erysiphe necator. 2019-2020 Host-Microbe Interactions Meeting. Davis, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lewis-Ivey, M. 2020. Fungicide Stewardship  Best Practices. FRAME Extension Meeting. Dublin, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Powdery Mildew Biology and Management . FRAME Extension Meeting. Dublin, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Best Sprayer Practices. FRAME Extension Meeting. Dublin, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Mahaffee W. 2020. Needles in haystacks: Inoculum monitoring as a decision aid. University of Florida, Dept. Plant Pathology. Gainesville, FL
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. The Past, Present and Future of Grape Pest Management. Ohio Grape Growers Conference. Dublin, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Brannen P. 2020. Fungicide and Insecticide Spray Program Workshop for Extension Agents. Georgia Extension Service. Blairsville, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Powdery Mildew Biology and Management . FRAME Extension Meeting. Prosser, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Oliver C. 2020. Fungicide Stewardship and Application Practices. FRAME Extension Meeting. Prosser, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Powdery Mildew: FRAMing the Reality of Fungicide Resistance. Business Enology Viticulture (B.E.V.) New York Annual Meeting. Rochester, NY
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Brannen P. 2020. West Georgia Wine Grape Spray Program Workshop. Carrol County Extension Service. Carrollton, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Brannen P. 2020. Northwest Georgia Wine Grape Spray Program Workshop. Lumpkin County Extension Service. Dahlonega, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Lewis-Ivey, M. 2020. QoI resistance in Ohio vineyards and management strategies to mitigate resistance development and the spread of resistance between vineyards. Ohio Winter Grape School. Madison, OH
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Miles T. 2020. Fungicide resistance research and mitigation strategies for grape powdery mildew . FRAME Extension Meeting. Napa, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. What makes for a challenging powdery mildew season? Environmental and cultural factors. FRAME Extension Meeting. Napa, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Oliver C. 2020. Decision-support tools for fungicide resistance management. FRAME Extension Meeting. Napa, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Brannen P. 2020. North Georgia Wine Grape Spray Program Workshop. Fannin County Extension Service. Ellijay, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sharma, N. 2020. Prebloom meeting on early season vineyard management and the importance of fungicide resistance. MSU Prebloom series. MI, (ZOOM)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Grape Disease Management  Powdery Mildew & Botrytis Bunch . Wine Island Growers Association. British Columbia (ZOOM)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Moyer M. 2020. Fungicide Resistance and the Acronyms. Pennsylvania Wine and Grape Team. Pennsylvania (ZOOM)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Pardini, C. 2020. Facing fungicide resistance in grape production: a game theoretic approach. AAEA Virtual Meeting. Virtually
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2020 Citation: Miles, T.D., Neill, T.M., Colle, M., Warneke, B, Robinson, G., Sterigiopoulus, I. and Mahaffee, W.F.(2020). Allele-specific detection methods for QoI fungicide resistant Erysiphe necator in vineyards. Plant Disease. https://doi.org/10.1094/PDIS-11-19-2395-RE In press
  • Type: Other Status: Published Year Published: 2019 Citation: Oliver C. 2019. FRAME Network: Fungicide Resistance in Wine Grapes. Washington State Grape Society. Grandview, WA (poster)
  • Type: Other Status: Published Year Published: 2019 Citation: Alzohairy, S., Sharma, N., Colle, M., Gillett, J., Sysak, R., Miles, T. 2019. 2019 update on fungicide resistance in Michigan vineyards. Great Lakes Fruit and Vegetable Expo. Grand Rapids, MI (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Oliver C. 2020. FRAME Network: Fungicide Resistance in Wine Grapes. Washington Association of Winegrape Growers. Kennewick, WA (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Oliver C. 2020. Grower Knowledge and Perception of Fungicide Resistance in US Vineyards. American Phytopathological Society. Virtually (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Warres, B., Neill, T., Mahaffee, W., and Brannen, P. 2020. Field efficacy of fungicides for management of grape powdery mildew in the presence of fungicide - resistant populations of Erysiphe necator in Georgia. American Phytopathological Society. Virtually (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Wong, A., and Mahaffee, W. 2020. A High Throughput Bioassay to CharacterizeErysiphe necator Tolerance to Multiple Fungicide Classes. American Phytopathological Society. Virtually (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Newbold, C., and Mahaffee, W. 2020. Phenotypic variability in the grape powdery mildew pathogen, Erysiphe necator. American Phytopathological Society. Virtually (poster)
  • Type: Other Status: Published Year Published: 2020 Citation: Lowder, S., and Mahaffee, W. 2020. Fits like a glove: Rapid, cost-effective sampling techniques to monitor Erysiphe necator. American Phytopathological Society. Virtually (poster)
  • Type: Other Status: Published Year Published: 2019 Citation: Moyer M., M. Cooper, P. Brannen, W. Mahaffee, M. Lewis-Ivey, T. Miles, and C. Oliver. 2019. Good to Know: Why Some Seasons are Worse for Powdery Mildew. Good Fruit Grower . https://www.goodfruit.com/why-some-seasons-are-worse-for-powdery-mildew/ (trade magazine)
  • Type: Other Status: Published Year Published: 2020 Citation: Miles T. 2020. Fungicide resistance a growing concern: Michigan grape growers must take steps to manage powdery mildew, botrytis bunch rot. Good Fruit Grower . https://www.goodfruit.com/fungicide-resistance-a-growing-concern/ (trade magazine)
  • Type: Other Status: Published Year Published: 2020 Citation: Lewis-Ivey, M. 2020. Managing Fungicide Resistance in the Vineyard. American Vineyard. https://americanvineyardmagazine.com/managing-fungicide-resistance-in-the-vineyard/ (trade magazine)
  • Type: Other Status: Published Year Published: 2020 Citation: Lewis-Ivey, M.2020. Managing Fungicide Resistance in the Vineyard. Ohio Grape Newsletter . https://ohiograpeweb.cfaes.ohio-state.edu/sites/grapeweb/files/imce/pdf_newsletters/March%202020%20OGEN.pdf (newsletter)
  • Type: Other Status: Published Year Published: 2020 Citation: Warres, B., and Brannen, P. 2020. Widespread QoI resistance and emergence of DMI resistance in powdery mildew in Georgia. Integrated Pest Management Newsletter. https://site.extension.uga.edu/ipm/2020/04/08/widespread-qoi-resistance-and-emergence-of-dmi-resistance-in-powdery-mildew-in-georgia/ (newsletter)
  • Type: Other Status: Published Year Published: 2020 Citation: Warres, B., and Brannen, P. 2020. Widespread QoI resistance and emergence of DMI resistance in powdery mildew in Georgia. Small Fruit News. https://smallfruits.org/2020/04/widespread-qoi-resistance-and-emergence-of-dmi-resistance-in-powdery-mildew-in-georgia/ (newsletter)
  • Type: Other Status: Published Year Published: 2020 Citation: Sharma, N., and Miles, T.2020. Grapevine powdery mildew fungicide resistance survey. MSU Extension News. https://www.canr.msu.edu/news/grapevine-powdery-mildew-fungicide-resistance-survey (newsletter)
  • Type: Other Status: Published Year Published: 2020 Citation: Tekip, A., and Miles, T.2020. Detecting emerging pesticide resistance in grapes. MSU AgBio Research. https://www.canr.msu.edu/news/detecting-emerging-pesticide-resistance-in-grapes (newsletter)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer M. 2019. The Past, Present, and Future of Viticulture Research and Extension at Washington State University. Washington State University - IAREC Centennial Celebration. Prosser, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Brannen P. 2019. Update on fungicide resistance in downy and powdery mildew diseases of grape in Georgia. Southeastern Professional Fruitworkers Conference. Tifton, GA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Stergiopoulos I. 2019. New developments in managing powdery mildew resistance. 8th Annual Vineyards & Wineries Continuing Education Class Series. Napa, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee W. 2019. Finding Needles in haystacks: Disease monitoring and risk assessment. National Grape Research Alliance. Sacramento, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee W. 2019. Needles in haystacks; Monitoring and mitigating fungicide resistance. University of California - Davis, Department Plant Pathology. Davis, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee W. 2019. Tracking fungicide resistant grape powdery mildew. Northwest Small Fruit Center Annual Conference. Ferndale, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Stergiopoulos I. 2019. Fungicide Resistance in grape powdery mildew across the Western US; What do we know so far?. FPS Grape Advisory Committee Meeting. Davis, CA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles T. 2019. Fungicide Resistance management in grapes: powdery mildew and botrytis. Great Lakes Fruit and Vegetable Expo. Grand Rapids, MI
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lowder S. 2019. Grape Powdery Mildew Management: Mildew's and Mildon'ts. FRAME Extension Meeting. Salem, OR
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer M. 2019. Best Sprayer Practices. FRAME Extension Meeting. Salem, OR


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

Outputs
Target Audience:The target audience for this project is broad and includes: grape growers, vineyards managers, vineyard interns, vineyard technicians, vineyard laborers, professional crop consultants, Extension professionals, plant health diagnostic clinics, chemical manufacturing companies, chemical distribution companies, industry organizations, undergraduate students, graduate students, and fellow researchers in the fields of viticulture, integrated pest management, plant pathology, fungicide resistance and disease epidemiology. We are reaching these audiences through surveys, on-farm and in-meeting demonstrations, workshops, and industry and extension publications (trade magazines, email blasts, social media posts, websites, and peer-reviewed extension publications).Rapid powdery mildew-sampling kits to screen for fungicide resistance have been made available to growers and consultants for the 2019 growing season. Sprayer coverage kits have been developed (in English and Spanish) to begin with initial training on what on-farm practices can lead to the selection of resistance, and how to correct for those actions. Nationwide surveys have been deployed to assess the general knowledge of growers, managers, and consultants on the presence of fungicide resistance; the responses from these surveys will be used to tailor and develop future extension and outreach efforts to target these specific audiences in the different geographical and cultural regions of the United States. Changes/Problems:We currently have no major changes within the scope of this project after the first 9 months. We did have a few delays in project hiring (postdoctoral scientists and graduate students). These were related to institutional delays in funding dispersement and account set-ups (most universities will not allow hiring of staff until accounts are fully established, which can take months after initial fund receival). The government shutdown over Dec 2018 - Feb 2019 significantly impacted two of our CoPIs during prime hiring time for students and postdocs related to project activities. We have since overcome these staffing delays, and have hired, or are in the process of hiring, the remaining budgeted staff and students associated with the project. What opportunities for training and professional development has the project provided?We (Extension team) delivered a full-day fungicide program design workshop, with 65 participants in Washington state, which provided additional opportunities for FRAME-associated postdocs and graduate students to participate and delivered pre-designed content, providing opportunities in public speaking and group-work with industry members. Postdocs and students have also engaged growers in OR and WA with specific demonstration workshops on how to properly scout for, and collect, grape powdery mildew. In addition, a YouTube instructional video was also made by a graduate student. Graduate students have also presented several posters and small research-based presentations (including short presentations at our annual project meeting) between September 2018 and May 2019. In addition to our postdoctoral research associates and graduate students, this project is providing training for1 undergraduate student, currently. How have the results been disseminated to communities of interest?Members of the Extension team (including graduate students) have delivered 35 FRAME-related presentations in multiple states since September 2018 with total cumulative attendance of over 1600 individuals. The audience type included both the industry and scientific community, with 31 of the 35 presentations targeted to industry. We have also presented several posters at regional grape conferences, and have had multiple articles shared via our university news feeds and trade magazines such as the Good Fruit Grower. We have also launched our project website at: framenetworks.wsu.edu, which will provide a holding ground for our durable outreach products. Social media accounts, such as Twitter and Facebook pages, have been established to help with the dispersal of materials. Additionally, regional grower targeted products such as blog posts, extension bulletins, and newsletters have been published in CA, GA, OR, MI, and WA. All articles and podcasts are featured on our FRAME project website. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: Extension and Outreach Obj. 1 - Activity 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United States. Plan of action - We plan on publishing our pre-project survey results in a peer-reviewed journal, highlighting key areas for targeting teaching and engagement. Obj. 1 - Activity 2: In-the-field demonstrations of management recommendations. Plan of action - Coordinated field demonstrations will begin in summer 2019, focusing on fungicide program design that incorporate rotation with non-risk products. After national surveying of the extent of fungicide resistance in 2019, these trials will be expanded in summer 2020 to reflect regional approaches to disease management. Obj. 1 - Activity 3: Develop a "University Service Center" business plan for fungicide resistance testing. Plan of action - No activity until years 3 and 4. Obj. 1 - Activity 4:Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Plan of action - We will continue to host regional workshops relating to basic powdery mildew management. We plan on writing additional extension articles on the basis of fungicide resistance and will work towards developing short videos and podcasts on basic fungicide resistance education. Objective 2: Group Decisions Obj. 2 - Activity 1:Conduct interviews to identify the externalities negotiating parties are experiencing (i.e., how their pesticide use affects their neighbors' production).Plan of action - All surveys have been distributed. Our objectives for the fall semester are: (1) analyze data, (2) literature review and (3) prepare a paper. Obj. 2 - Activity 2: Use game theory to develop compensating mechanisms that induce cooperation and aid fungicide stewardship by inducing cooperative behavior among growers, crop consultants and chemical companies. Plan of action - No intended activity until after year 2. Objective 3: Detection and Monitoring Sub-Objective 3.1-Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Activity 1:Development of PCR based diagnostic assays.Plan of action - Complete FRAC 11 detection manuscript and submit for peer reviewed publication. For SDH mutations that increase the most tolerance of the fungus against SDHIs, TaqMan and other PCR-based diagnostic assays will be developed and tested for specificity and sensitivity. High resolution melting analysis will also be pursued as a method to simultaneously detect multiple mutations in the SDH genes. Obj. 3.1- Activity 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Plan of action -Continue to optimize isothermal assays for sensitivity and specificity and conduct technology transfer experiments with other partner laboratories. Obj. 3.1 - Activity 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Plan of action - We will continue to monitor the incidence and dynamics of SDH gene mutations bytarget-site gene sequencing in collected samples from multiple states,and determine the effect of these mutations on SDHI tolerance.A multiplex PCR assay is being developed that will allow us to amplify the three SDH genes in a single PCR reaction and subsequently sequence them with gene-specific primers. Sub-Objective 3.2-Monitoring for Fungicide Resistance. Obj. 3.2 - Activity 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Plan of action - Field collections are currently under way for the Spring 2019 growing season. In CA, commercial vineyards will be monitored for fungicide resistance by surveying powdery mildew colonies and through a network of fixed, rotorod spore traps. In OR, we are monitoring for E. necator and fungicide resistance by deploying 25 spore traps in the region. In OR, MI, GA, and WA, vineyards will be intensively sampled using leaf, glove, and isolate sampling to examine which method is the efficient for allowing an accurate grower decision. This sampling will be in collaboration with a large consulting group to cover more vineyards on the west coast. Obj. 3.2 - Activity 2:Phenotype for fungicide sensitivity.Plan of action - We will collaborate with 3 growers that quit using DMI and QoI fungicides in 2016 and have lower or no detectable QoI resistance populations to see if QoIs and DMI fungicides can be rotated back into their program. We will continue to bioassay individual isolates to confirm molecular resting and identify new resistant phenotypes. Sub-Objective 3.3-Understanding the Basis of Resistance. Obj. 3.3 - Activity 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides. Plan of action - Populations will start to be developed in year two, with activity focus occuring in year 3 and 4. Obj. 3.3 - Activity 2:Create reference genome ofE. necatorfor comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Plan of action - A combined assembly will be generated using two draft assemblies produced from DNA long-reads and Hi-C data. The final assembly will be polished and completeness will be assessed using a minimum of five standard metrics, including N50 and L50 measures. If necessary, additional long-read sequences will be pursued by Nanopore sequencing, using theUC Davis genome center. After full assembly, transcripts will be annotated during the winter of 2019, finishing the process and publishing the genome in early 2020. Obj. 3.3 - Activity 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis.Plan of action - We are currently in the process of hiring another graduate student to continue work on FRAC 7 (SDHI) fungicide resistance. Objective 4: Predicting Resistance Spread Sub-Objective 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Activity 1:Sample collections and phenotyping. Plan of action - A postdoctoral research candidate has been identified to work on the project starting in July, and federal clearance paperwork has been initiated. Sampling from each location will continue through the end of the grape growing season in each location. Obj. 4.1 - Activity 2: Genome sequencing and allelic diversity analyses. Plan of action - Initial population sequencing will begin in year 2, with activity focus in years 3 and 4. Sub-Objective 4.2- Dispersion Tracking and Population Prediction. Obj. 4.2 - Activity 1.1:Mapping resistant isolate spread risk. Plan of action - A postdoctoral research candidate has been identified to work on the project and a formal offer has been extended to the candidate. Construction of model domains for each FRAME test vineyard will commence throughout the summer and downscaled weather forecasts in both a reanalysis and forecast model will be performed and archived. Obj. 4.2 - Activity 1.2: System validation and evaluation. Plan of action - Major activity not planned until after year 2. Sub-Objective 4.3-Fungicide Record Evaluation. Plan of action - Collection of fungicide records will begin during the spring and will continue through the fall 2019. Data analysis of records will begin in winter 2019, with preliminary models comparing use patterns to time of noticeable control failure and resistance detection developed by end of year 2. Model refinement and validation in years 3 and 4.

Impacts
What was accomplished under these goals? At the time of reporting, we are currently only 9 months into the project, and just starting the first in-field season. Not all project goals and objectives were slated for the first year. Updates for what was completed / in progress for Year 1 are below: Overall accomplishments - The first annual project meeting for FRAME was held on 27 Feb - 1 Mar, 2019, in Prosser WA. Participants included all project PIs and Key Personnel,Stakeholder Advisory group, technicians and students. Objective Teams hosted regular web conferences from weekly to quarterly, depending on nature of the objective). The project website (https://framenetworks.wsu.edu/), was developed. Objective 1: Extension and Outreach Obj. 1 - Activity 1: Pre- and post-project surveys to track short-term learning, knowledge, and skills changes in grape growers across the United States.Year 1 accomplishments - Completed our nation-wide pre-project survey, designed to gather regional baseline understanding of fungicide resistance among grape growers. Data analysis to begin in late summer 2019. Obj. 1 - Activity 2: In-the-field demonstrations of management recommendations. This is just starting for 2019. Obj. 1 - Activity 3: Develop a "University Service Center" business plan for fungicide resistance testing. Not scheduled until years 3 and 4. Obj. 1 - Activity 4:Develop workshops and other classic extension tools; create and engage with enhanced local and regional industry groups.Year 1 accomplishments - Members of the Extension team delivered 35 FRAME-related presentations since Sept. 2018 to a cumulative 1600 individuals. The audience type included both the industry and scientific community, with 31 of the 35 presentations targeted to industry. There were several student posters, a pod cast, and several feature articles on the project between September 2018 and May 2019. All articles and podcasts are featured on the FRAME website. Social media accounts have been established to assist in information "push". Objective 2: Group Decisions Obj. 2 - Activity 1:Conduct interviews to identify the externalities negotiating parties are experiencing (i.e., how their pesticide use affects their neighbors' production).Year 1 accomplishments -We developed three surveys which were distributed (Growers Survey, Crop Consultant Survey) and May (Manufacturers survey). The Growers Survey aims to identify the level of knowledge about fungicide resistance among grape growers and their willingness to adjust their own practices if it meant a regional benefit in mitigate crop loss due to fungicide resistance. The Crop Consultant Survey seeks to determine the level of knowledge about fungicide resistance among crop consultant and their willingness to adjust their sales and program designs if it meant a regional improvement in management of fungicide resistance. The Manufacturers Survey will evaluate what changes in production or marketing manufacturers would deem appropriate in light of potential product efficacy loss due to resistance. Obj. 2 - Activity 2: Use game theory to develop compensating mechanisms that induce cooperation and aid fungicide stewardship by inducing cooperative behavior among growers, crop consultants and chemical companies. Not scheduled until after year 2. Objective 3: Detection and Monitoring Sub-Objective 3.1-Develop Molecular Markers to Improve the Speed of Resistance Detection. Obj. 3.1 - Activity 1:Development of PCR-based diagnostic assays.Year 1 accomplishments - An improved rapid sampling technique was developed that reduced sampling kit manufacturing time by 80% and the time to collect sample by 60%, with no effect on sample processing time or sensitivity. We refined the y136F qPCR assay for detecting Y136F mutation associated with FRAC 3 resistance, including correlating Cyp51 copy number in relation to genome copy number (and subsequent expression of FRAC 3 resistance phenotype). Obj. 3.1- Activity 2: Developing isothermal diagnostic tools for in-field detection of fungicide resistance. Year 1 accomplishments - We developed an isothermal molecular diagnostic marker for FRAC 11 resistance, which should allow for field detection of fungicide resistance. Obj. 3.1 - Activity 3:Target-site gene sequencing to diagnose DMI, QoI, and SDHI resistance.Year 1 accomplishments - Genetic libraries were developed to study FRAC 13 resistance and to examine the transcriptome profile of FRAC 13 sensitive and resistant isolates. Libraries are actively being created for examination of SDHI resistance profiles to screen for consistant mutation prevalence and relavence to field resistance. Sub-Objective 3.2-Monitoring for Fungicide Resistance. Obj. 3.2 - Activity 1:Assess the sampling density and pattern needed to estimate proportion of E. necator with fungicide resistance.Year 1 accomplishments - We standardized sampling protocols across collaborators in CA, GA, MI, OR, and WA; it included three experimental sampling proceedures and research plot design. Obj. 3.2 - Activity 2:Phenotype for fungicide sensitivity.Year 1 accomplishments - A germination bioassay for FRAC 3, 9, 11, 13, 50 (formally U8) and U6 fungicides was develop that improved throughput by >400%. Sub-Objective 3.3-Understanding the Basis of Resistance. Obj. 3.3 - Activity 1: Elucidate mechanisms of resistance development by experimentally evolving populations with resistance to SDHI and AzN fungicides. Just starting this field season. Obj. 3.3 - Activity 2:Create reference genome ofE. necatorfor comparative genomic and transcriptomic approaches to identifying fungicide resistance mechanisms.Year 1 accomplishments - We developed an improved procedure for growing E. necator to increase DNA concentrations for Hi-C or PacBio sequencing. We have produced over 20mg of E. necator E1-101 DNA for sequencing, meeting Objective target 9 months ahead of schedule. A current draft of a genome is under construction using 3 separate methods for validation and consensus comparison. Obj. 3.3 - Activity 3: Identification of genomic markers associated with fungicide resistance through comparative genome and transcriptome analysis. Not slated year 2. Objective 4: Predicting Resistance Spread Sub-Objective 4.1:Conduct seasonal monitoring and recurring collection of isolates from selected grower vineyards for AzN, DMI, QoI and SDHI resistance and genetic diversity. Obj. 4.1 - Activity 1:Sample collections and phenotyping. Year 1 accomplishments - Selection of field sampling locations and physical sample collectionsfor the 2019 season (first full growing season) are underway. Obj. 4.1 - Activity 2: Genome sequencing and allelic diversity analyses. Not slated until after year 2. Sub-Objective 4.2- Dispersion Tracking and Population Prediction. Obj. 4.2 - Activity 1.1:Mapping resistant isolate spread risk. Year 1 accomplishments - A model for airborne isolate spread at the vineyard scale was developed and validated prior to the project's start. Further tests downscaling forecasted regional weather to the vineyard scale (~1 km) where performed. These initial tests downscaled forecasts from the North American Mesoscale Forecast System (NAM) at 12 km to 1 km using the Weather Research and Forecasting (WRF) model. Obj. 4.2 - Activity 1.2: System validation and evaluation. Not slated until after year 2. Sub-Objective 4.3-Fungicide Record Evaluation. Year 1 accomplishments - We began soliciting fungicide records from participating growers at the start of the 2019 growing season (April).

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles, T. (2019). Dormant and early season management including information about "softer chemicals". presented at MI Statewide Pre-Season Kickoff meeting, Benton Harbor, MI.
  • Type: Websites Status: Published Year Published: 2018 Citation: https://framenetworks.wsu.edu/
  • Type: Other Status: Published Year Published: 2019 Citation: Lowder, S. (2019). Rapid sampling techniques to monitor group 11 fungicide resistant grape powdery mildew. Poster presentation to grower groups, Corvallis, OR.
  • Type: Other Status: Published Year Published: 2019 Citation: Newbold, C. (2019). Succinate dehydrogenase inhibitor resistance in isolates of Erysiphe necator from the Western U.S. Poster presentation to grower groups, Corvallis, OR.
  • Type: Other Status: Published Year Published: 2019 Citation: Wong, A. (2019). A High Throughput Bioassay to Rapidly Characterize Erysiphe necator Resistance to Demethylation Inhibitor Fungicides. Poster presentation to grower groups, Corvallis, OR.
  • Type: Other Status: Published Year Published: 2019 Citation: Neill, T. (2019). An International Mildew Management Problem  fungicide resistance from coast to coast. Poster presentation to grower groups, Corvallis, OR.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2019 Citation: Colle., M. Neill, T., Mahaffee, W., Miles, T. (2019). Development of a rapid isothermal assays to detect QoI resistance in Erysiphe necator the causal agent of grape powdery mildew. Phytopathology Abstr.
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2019 Citation: Sharma, N. Neill, T., Mahaffee, W., Miles, T.D. (2019). Development of a rapid isothermal assays to detect QoI resistance in Erysiphe necator the causal agent of grape powdery mildew. Phytopathology Abstr.
  • Type: Journal Articles Status: Other Year Published: 2020 Citation: Colle, M., Neill, T., Warnke, B., Mahaffee, W., Miles, T.D. (201X). Detection of Erysiphe necator fungicide-resistant alleles in leaf and air samples using novel molecular diagnostic techniques. (in preparation, submitting to Plant Disease).
  • Type: Other Status: Published Year Published: 2019 Citation: Hansen, M., and M. Moyer. 2019. Basic Training for Combating Mildew. Wine Business Monthly. March 2019: 84  91. Online: https://www.winebusiness.com/wbm/
  • Type: Other Status: Published Year Published: 2019 Citation: Oliver, C. 2019. Avoiding Selection of Fungicide Resistance. WSU Viticulture and Enology Extension News - Spring 2019. Online: http://wine.wsu.edu/extension/viticulture-enology-news-veen/
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles, T. (2019). Grape Disease Management and Program Directions. presented at Southwest Horticulture Days, Benton Harbor, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles, T. (2019). How Fungicide Resistance Occurs, and Using Resistance Management Concepts in Mildew Management Programs. presented at FRAME Extension Meeting, Prosser, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles, T. (2019). Vineyard Integrated Pest Management: Diseases. presented at MI Statewide MSU Grape School, Benton Harbor, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Mahaffee, W. (2018). Fungicide Resistance in grape production. presented to BASF representatives, Raleigh, NC.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Mahaffee, W. (2018). Fungicide resistance, selection, and timing. presented at the Sustainable Ag Expo, San Luis Obispo, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Mahaffee, W. (2018). Fungicide stewardship. presented to PCA representatives, Woodland, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Mahaffee, W. (2018). Grape powdery mildew management in the era of fungicide resistance. presented to the Growest PCA conference, Woodland, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee, W. (2019). What the mildew?. presented at the Lodi Grape Growers Conference, Lodi, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee, W. (2019). Fungicide resistance, selection, and timing. presented to Nutrien Crop Consultants, McMinnville, OR.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Mahaffee, W. (2019). Fungicide resistance, selection, and timing. presented to North-Coast CAPCA, Santa Rosa, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Robinson, G. (2019). Monitoring of QoI and SDHI Resistance in Grape Powdery Mildew through Target-Site Mutation Detection  A Critical Update. presented to UC Davis Extension via Current Wine and Winegrape Research Course, Davis, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Stergipopoulos, I. (2019). An update on SDHI resistance monitoring and the (mt) genome of Erysiphe necator. presented to FRAME research team, Prosser, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Moyer, M. (2018). Realizing the Role of People in Grape Disease Management. presented to WSU Dept of Plant Pathology, Prosser, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Moyer, M. (2018). Grape Powdery Mildew Update. presented to Washington State Grape Society, Grandview, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Monitoring and Mapping Grape Powdery Mildew Fungicide Resistance and Crown Gall Incidence in Washington Vineyards. presented to Washington State Grape and Wine Research Program, Prosser, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Fungicide Management. presented to Washington Winegrowers Association, Kennewick, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Managing Mildew and Fungicide Resistance: Back to Basics. presented to Gallo Grape Growers - South Valley, Fresno, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Managing Mildew and Fungicide Resistance: Back to Basics. presented to Gallo Grape Growers - North Valley, Lodi, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Designing a Mildew Spray Program. presented during FRAME extension meeting, Prosser, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Managing Crown Gall and Powdery Mildew Diseases in the Columbia Gorge. presented to WAVEx (Washington State Wine Commission), Dalles, OR.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moyer, M. (2019). Powdery Mildew Management in Concord - An Update. presented to National Grape Cooperative Growers, Grandview, WA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Cooper, M. (2018). Powdery Mildew Fungicide Resistance Update. presented at UCCE Lake/Mendocino IPM Conference, Hopland, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Cooper, M. (2018). Strategies for managing fungicide resistance. presented to Napa Regional Grower Group, Oakville, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Cooper, M. (2019). Monitoring fungicide resistance with a spore trap array in Napa County. presented to Napa Regional Grower Group, Oakville, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Brannen, P. (2018). Grape Post Harvest Round Table. presented to Georgia Wine Grape Producers, White, GA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Brannen, P. (2018). Fungicide Sensitivity Profiling of Grapevine Downy Mildew in Georgia. presented at Southeastern Professional Fruit Workers Conference, Manchester, TN.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Brannen, P. (2019). Powdery and Downy Mildew Resistance Update. presented to Georgia Wine Growers Association, Braselton, GA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Brannen, P. (2019). Powdery and Downy Mildew Resistance Updates. presented to Georgia Wine Grape IPM Meeting, Elijay, GA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lowder, S. (2019). Rapid methods for monitoring fungicide resistance in grape powdery mildew. presented at Raisin and Table Grape Growers annual Conference, Visalia, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lowder, S., Neill, T., Moyer, M., Miles, T., Stergiopoulous, I., and Mahaffee, W. (2019). Monitoring fungicide resistant grape powdery mildew and the use of micronized sulfur. California Table Grape Commission Conference, Visalia, CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Miles, T. (2018). Fungicide resistance and viruses commonly found in Michigan. presented at Great Lakes Fruit and Vegetable Expo, Grand Rapids, MI.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Miles, T. (2019). Detecting viruses and fungicide resistant pathogens in MI vineyards. presented at Orchard and Vineyard Show, Traverse City, MI.