Source: MICHIGAN STATE UNIV submitted to
A PRODUCTION SYSTEM FOR HIGH VALUE CROPS AT RISK FROM DOWNY MILDEW: INTEGRATING DETECTION, BREEDING, EXTENSION, AND EDUCATION.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1008855
Grant No.
2016-68004-24931
Project No.
MICL08516
Proposal No.
2015-08750
Multistate No.
(N/A)
Program Code
A5160
Project Start Date
Mar 1, 2016
Project End Date
Feb 28, 2021
Grant Year
2018
Project Director
Hausbeck, M.
Recipient Organization
MICHIGAN STATE UNIV
(N/A)
EAST LANSING,MI 48824
Performing Department
Plant Soil Microbial Sciences
Non Technical Summary
Downy mildews are devastating diseases that infect many vegetable crops important to healthy diets (lettuce, spinach, cucumber, melon, pumpkin, squash, onion, broccoli, cabbage) and valuable local and export crops (hop, basil, sunflower) that are worth $7.5 billion yearly to U.S. growers. Our team proposes a team-based scientific approach to develop a growing system that limits the destructive effects of these downy mildew diseases. We will (1) use DNA and molecular biology to quickly identify downy mildew disease of cucumber, melon, pumpkin, squash, lettuce, spinach and basil, and determine if it is resistant to fungicides, (2) track downy mildew disease of cucumber, melon, pumpkin, squash and basil yearly on a map at a website that growers and home gardeners can access, (3) breed basil, spinach and cucumber plants that are resistant to downy mildew, (4) evaluate whether our research solutions will be economical, (5) inform growers and the public of our research, and (5) involve college students in our research so they can be better prepared for jobs once they graduate. Growers will limit the losses from downy mildew diseases of cucumber, melon, pumpkin, squash, lettuce, spinach and basil by making informed decisions based on tested solutions developed through science. We will educate diverse and underrepresented college students in agricultural sciences. The solutions from our research will apply to other crops affected by other types of downy mildews because the pathogens that cause these diseases are similar. Therefore, this project will also benefit a wide group of growers and consumers interested in onion, sunflower, hop, broccoli, and cabbage.
Animal Health Component
0%
Research Effort Categories
Basic
70%
Applied
30%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2161421116015%
2161420116015%
2161430116010%
2165220116010%
7115220116010%
2021421108010%
2021430108010%
2161421117010%
2051421117010%
Goals / Objectives
The goal of our Coordinated Agricultural Project (CAP) is to improve/develop novel Agricultural Production Systems that detect and track downy mildew (DM) pathogens and contain them through sustainable and effective mitigation. To accomplish this we will focus on the following food crop/DM pathogen combinations: cucurbits/Pseudoperonospora cubensis, spinach/Peronospora effusa, lettuce/Bremia lactucae, and basil/Peronospora belbahrii.OBJECTIVE 1. Early detection and rapid diagnostics: We will improve early detection by developing rapid diagnostics for DM that will alert growers of pathogen influx/overwintering, and distinguish among DM species. A DM database with diagnostic tools will be generated and hosted at the unified project website proposed in Objective 2.1i) Species-specific genetic markers for diagnostics.1ii) Diagnostic assays for early detection: cucurbits, lettuce, spinach, basil.1iii) Fungicide-resistance diagnostics: lettuce spinach, basil.OBJECTIVE 2. Tracking pathogen dispersal and survival: We will prevent and contain specific DM pathogens by establishing a website that will track DM outbreaks nationwide, provide data that will support optimal timing of fungicide applications, quantify fungicide activity, report overwintering sources, and provide control recommendations.2i) Expand the national DM tracking website: cucurbits, basil.2ii) Deploy molecular diagnostics: spore trapping cucurbits, basil; field test cucumber, spinach, lettuce, basil.2iii) Provide grower alerts based on DM inoculum strength: cucurbits.2iv) Determine DM survival: cucumber, melon.2v) Quantify duration of disease control with DM fungicides: cucumber, watermelon, cantaloupe, basil.OBJECTIVE 3. Developing DM-resistant crops: We will develop improved resistant basil, spinach, and cucumber germplasm.3i) Develop DM-resistant crop germplasm: basil, spinach, cucumber.3ii) Participatory plant breeding of DM-resistant: cucumbers.OBJECTIVE 4. Cost/benefit ratios and grower adoption: We will compare economic costs, returns and risks of grower acceptance and adoption of strategies including a cost analysis of new, resistant germplasm, and DM prevention, diagnostic, and forecasting systems.4i) Baseline DM disease impacts to food security.4ii) Economic impact of the behavioral, social, and environmental outcomes.OBJECTIVE 5. Outreach and adoption: We will conduct a nationwide outreach program to increase adoption of DM mitigation strategies and increase fungicide efficiency. We will host a national conference for researchers, extension educations, and producers. The effectiveness of our outreach activities will be evaluated.5i) DM monitoring/forecasting systems.5ii) Deployment of DM-resistant breeding lines/varieties.5iii) Seed testing for basil DM.5iv) Field spray recommendations.5v) National conference.5vi) Social media.OBJECTIVE 6. Attract students to agricultural sciences: We will provide research and extension education opportunities to undergraduate students via summer internship programs.
Project Methods
OBJECTIVE 1. Early detection and rapid diagnostics. Nuclear and mitochondrial diagnostic markers of Pseudoperonospora cubensis (cucurbits DM), P. effusa (spinach DM), B. lactucae (lettuce DM), P. belbahrii (basil DM), will be used to develop and deliver assays that can detect DM in spore trap samples and infected seed. Candidate diagnostic markers will be tested using a multiplex real-time polymerase chain reaction (PCR) technique. DM spore numbers will be correlated with the real-time PCR, and validated with isolates from multiple hosts and geographic regions. Diagnostic assays for the carboxylic acid amide, benzimide and QoI fungicides for which the genetic basis of resistance is currently known in P. cubensis will be developed and tested for transferability to the other DM pathogens, including those associated with lettuce, spinach, and basil. Assays will be developed in platforms that can be adopted by Plant Diagnostic Clinics and extension personnel and include PCR-based assays and isothermal recombinant polymerase amplification assays.OBJECTIVE 2. Tracking pathogen dispersal and survival. The current cucurbit ipmPIPE DM tracking website (http://cdm.ipmpipe.org) will be expanded to include basil DM. Molecular diagnostic tools will be validated by comparing the tools' quantitative PCR (qPCR) methods to conventional spore trapping methods and risk maps developed based on a spore trap network. Tools will test presence of DM in basil and cucurbit seeds. Weekly data and landscape level analyses will quantify the source strength of an infected cucurbit crop with extent of final DM spread and severity. This will be used in conjunction with weather, host susceptibility and other risk factors to develop a predictive model to advise growers when DM infection is imminent and management techniques need to be implemented. The possibility of oospore production in regions with a frost/freeze period will be investigated by inoculating cucurbit fields with isolates of opposite mating types and evaluating for oospore presence. Pre- and post-infection duration of disease control from DM fungicides compared to untreated plots will be quantified in the field and modeled to provide fungicide selection and timing recommendations for the ipmPIPE website.OBJECTIVE 3. Developing DM-resistant crops. Diverse germplasm of basil, spinach and cucumber will be screened for DM resistance and potential resistance gene(s) will be introduced into marketable cultivars and varieties by crossing and recurrent selection for DM resistance and desirable marketable traits. Participatory plant breeding will be established with cucumber growers in Michigan, North Carolina and New York to recurrently select DM-resistant cucumbers from a population of cucumber germplasm.OBJECTIVE 4. Cost/benefit ratios and grower adoption. A cost-benefit model will be developed to capture the cost and benefits associated with each of the innovative methods to control DM. An economic analysis of field experiments will be performed to compare, with partial budgeting, the expected net returns and cost of production for each of the innovative and traditional methods. All input and output prices will be standardized to focus comparison on yield and input use differences. Using secondary sources, price data will be collected. The model will integrate the experimental data with economic input and output prices to determine the economic returns for DM management strategies. Yield data comparisons from differing control strategies will be critical for this assessment. Additional information relating to the economic impact of DM will be collected through focus group meetings of experts. A questionnaire including questions relating to production input, alternative methods to control DM, cost and returns associated with each alternative method, insecticide and fungicide applications and demographics characteristics will be designed for producers, based on published literature on the subject as well as experiences of the co-PIs in the field.OBJECTIVE 5. Outreach and adoption. The national reporting/disease forecasting and monitoring website for cucurbit DM will be updated to include basil DM. Demonstration plots will introduce new breeding lines/varieties of cucumber, spinach and basil to local growers during field days. Diagnostic tools will be tested and provide information on potential basil seed contamination to growers. Best Management Practice guides for fungicides will be developed on basil, lettuce and spinach DM and disseminated at local, state, regional and national meetings and published online. A national conference will be scheduled to disseminate results to researchers, extension educators, industry personnel, and growers. A web presence will be created through project profiles at networking sites such as LinkedIn, Facebook and Twitter, and the project's sites will be promoted through project PIs' websites and social media.OBJECTIVE 6. Attract students to agricultural sciences. A summer undergraduate research program for 10-12 students will be implemented to serve two demographics: Hispanic serving institutions (HSI) and multi-campus internships in various laboratories of other team members. Additionally, we will offer 3 guest lectures in a Plant Pathology course taught at an underserved HSI campus, which will impact 20-30 students per year.

Progress 03/01/16 to 02/28/21

Outputs
Target Audience:Our target audience includes people in the scientific, extension, and agricultural communities. Included are other scientists, undergraduate and graduate students, and post-docs in plant sciences, plant breeding and plant pathology, crop consultants, extension agents, crop protection industry, growers, shippers, processors, and other allied stakeholder industries. Changes/Problems:Due to the COVID-19 pandemic, co-PI Smart was not able to conduct field trials or host an undergraduate in 2020. Other project field trials and lab work were amended due to the limitations of COVID-19 safety restrictions on travel to field sites and the need to stagger personnel in the lab. Result dissemination was done in a virtual format in 2020. These changes did not affect completion of the main objectives of this project. What opportunities for training and professional development has the project provided?This project has resulted in the training of many undergraduates, graduate students, and post-doctoral scientists in both field- and laboratory-based agricultural scientific techniques. Field-based techniques include conventional and q-PCR, Burkard volumetric spore trapping and rotorod spore trapping, weather monitoring, establishing and evaluating greenhouse and field fungicide/biopesticide efficacy trials, sentinel plots, and cultivar trials including cucurbit, lettuce, spinach, and basil production, field plot design, establishment, seed germination, transplanting, maintenance, fungicide application, disease diagnostics/PCR diagnostics, disease evaluation, yield assessments, canopy cover imaging and analysis, greenhouse plant production, cucumber pollinations and seed processing, cross and self-pollination and drying of basil, organizing breeding populations in the field, and prioritizing selections. Laboratory-based techniques include DM pathogen sampling and sporangial and DNA preservation, transfers of DM onto new host plant material and cell counting; growth and storage of P. cubensis and P. humuli, DNA extraction, PCR, qPCR, SSR/microsatellite genotyping sequencing analyses and Sanger sequencing; design and development of diagnostic assays including the mitochondrial-based TaqMan Diagnostic assay. Other techniques learned included statistics, network modeling approaches in biological systems, image analysis techniques for disease damage quantification, biomathematics on network modeling of cucurbit DM, genomics, bioinformatics, epidemiology, the role of oospores in the pathogen survival and epidemiology, designing, developing and testing of the website for tracking basil DM, coordination and compilation of grower survey results, and preparation of presentations, technical reports, and manuscripts. Undergraduates (33) were accepted into Research Experience for Undergraduates (REU) sites at North Carolina State Univ., Cornell Univ.-Geneva, Cornell Univ.-Boyce Thompson Institute, Pennsylvania State Univ., and Michigan State Univ. through Co-PI Miles's program. Post-doctoral scientists, graduate students, and undergraduate students have been trained in the methods and techniques listed above as required for this project under the supervision of the PD and co-PIs and aided by technicians. Those who have benefited from this training include post-doctoral scientists (Safa Alzohairy, Surendran Arumugam, Elizabeth Brisco, Alyssa Burkhardt, Shyam Kandel, Alamgir Rahman, Andres Salcedo), graduate students (Julian Bello, Lara Brindisi, Lauren Brzozowski, Kim D'Arcangelo, Phong Dang, Sara Getson, Juan Gonzalez Giron, Katelyn Goldenhar, Lexi Heger, Doug Higgins, Kathryn Homa, Grace Kenny, Robert Mattera, Anthony Noto, Maureiq Ojwang, David Perla, Robert Pyne, Trevor Ruiz, Andy Shirey, Martha Sudermann, Samantha Thompson, Matthew Uebbing, Greg Vogel, Tianxin Wu, Jesse Yamagata), undergraduate students (Andrew Abdelmalek, Carson Adams, Madison Ahmad, Andrew Aldcroft, Stephanie Aponte, Sierra Barnes, Tanner Boss, Hyacinth Burrows, Ali Cala, Baylee Carr, Jordan Castellari, Jia-Yi Chan, Brian Change, Howard Chhen, Samuel Cude, Gina Dabbah, Maeve Day, Phi Diep, Abby Dooley, Jacob Dorn, Paul Anthony Edwards, Philip Engelgeau, Morgan Gifford, Amber Glenn, Ryan Gould, Asia Green, William Gura, Jolie Habashy, Ross Hatlen, Lexi Heger, Emily Henderson, Madeline Hendrickson, Clayton Jarvis, Thilani Jayakody, Adam Job, Kavindi Karunaratne, Nicholas, King, Elizabeth Kogan, Krupali Kathari, Jacob Kowaleski, Courtney Lepping, Devin Linderman, Adrian Maldonado, Anastasia Matta, Vicki Meraz, Laura Merja, Kelly Merrill, Sara Merrill, Reilly Nakomoto, Han Nguygen, A.J. Noto, Lisa Panichelli, Gabriel Ramirez, Benjamin Rouse, Carly Seldow, Aidan Shands, Trevor Styles, Halley Taddonio, Opal Tam, Jessica Tan, Andrea Karina Suazo, Jonathan Vantman, Miranda Weatherly, Darius Wells, Yazmin Williams, Danielle Wolleman, Jesse Yamagata, Li Yang), and technicians (Mariami Bekauri, Tanner Boss, Matt Bour, Alyssa Burkhardt, Briana Claasseen, Alex Cook, Uma Crouch, Colin Day, Christopher DelCastillo, Allison DeSario, Cheryl Engfehr, Jack Fabrizio, Garrett Giles, Asia Green, Blair Harlan, Taylere Herrmann, Barry Hughes, Scott Hughes, Gregory Inzinna, Marc Jaquez, Steve Kalb, Saben Kane, Rachel Kreis, Emi Kuroiwa, John Kurtz, Holly Lange, Sheila Linderman, Macus Lopez, Patricia Love, Leah Martin, Mike Matson, Evan McClenthan, Marina Ramon, Paige Reeves, Emily Rustad, Joe Shemanski, Brian Sheppard, Sheri Smith, Shane Sullivan, Rebecca Willis, Sascha Zepeda, Richard Zimmerman). How have the results been disseminated to communities of interest?Research results have been disseminated via grower demonstration plots, one-on-one grower conversations and farm visits, publications, websites, scientific/extension presentations, webinars, and scientific/extension meetings. NIFA was acknowledged in 104 presentations. Scientific Meetings: Results were disseminated to the scientific community in 69 presentations at scientific meetings, including yearly presentations at the Annual Meeting of the American Phytopathological Society, Cleveland, OH. Extension Meetings: Research results were disseminated as extension presentations at 146 meetings at the local, regional, and national levels, to growers, commodity groups, chemical companies, extension educators, and other agricultural industry stakeholders. Meetings included yearly presentations at the Pickle Packers International Annual Meeting, Great Lakes Farm, Fruit and Vegetable Expo, NJ-NOFA Annual Conference, California Leafy Greens Research Program (CLGRP) Annual Meeting, Eastern New York Fruit & Vegetable Conference, Western New York Vegetable Growers Meeting, Empire State Producers Expo, Syngenta Annual Research Meeting, Citrus Expo and Vegetable & Specialty Crop Expo, North Carolina Vegetable Growers Association Ag Expo, Agent Training on Disease Diagnostics and Management in Vegetable Crops, and numerous yearly local extension meetings. Webinars, open houses, workshops, field days, trial tours: Fifty-three webinars, open houses, workshops, field days, and trial tours featured this project's research, including the annual USDA NIFA Basil Workshops, and Phytophthora and Cucurbit Downy Mildew Workshops. Five articles featuring project research, authored by people other than project PD/co-PIs, were published in Edible Jersey, GrowerTalks, Greenhouse Grower, Inside Grower, and Futures. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1. Co-PI Gent developed assays (3) to differentiate P. cubensis vs. P. humuli in airborne sporangia and leaf tissue. Co-PI Gent provided P.humuli DNA and isolates to PD Hausbeck and Co-PIs Gent, Quesada, and Vallad to test assays. Co-PI Quesada identified species-specific nuclear diagnostic markers and developed assays for P. cubensis clade 1 and 2 isolates. Co-PI Martin developed mitochondrial markers for detection of P. cubensis (clade 1 and 2) and P. humuli and validated markers to determine the relationship between spore counts and PCR results. Co-PI Smart compared assays from co-PIs Martin and Quesada. A mitochondrial assay developed by Co-PI Smart was tested; the assay developed by Co-I Martin was quicker and easier to perform. PD Hausbeck added an internal control to the qPCR assay developed by Co-PI Martin to lower the quantification limit to 3 sporangia. Co-PI Vallad evaluated clade-specific P. cubensis markers provided by Co-PI Quesada against 274 isolates. PD Hausbeck used spore traps and the qPCR assay to detect P. cubensis in MI. Co-PIs Miles and Quesada developed a qPCR assay for fungicides, targeting FRAC codes 11 and 40. PD Hausbeck tested the assay for MI P. humuli populations. Efforts were focused on SNP-based TaqMan markers due to their easy transferability to diagnostic clinics. Objective 2. Sentinel plots for DM were established yearly in MI, NY, FL, NC, and NJ. Burkard volumetric, ionic volumetric, and impaction traps were used in FL by Co-PI Vallad. Co-PI Smart deployed roto-rod traps but PCR assays were inconsistent. PD Hausbeck used Burkard volumetric and impaction traps in MI fields. DNA of P. cubensis and P. humuli were detected by Burkard traps, while only P. cubensis DNA was detected by impaction traps. qPCR results from Burkard trap samples were more closely correlated with spore counts from light microscopy when sporangial counts were low, compared to impaction traps. Confirmed DM cases were reported to the CDMipmPIPE and the Basil Ag Pest monitor, and disseminated via websites and bulletins. Diagnostic assays for P. cubensis clade 1 and clade 2 isolates were developed and validated. Assays to detect to QoI and CAA fungicide resistance mutations were developed and tested. Species-specific nuclear diagnostic markers for P. belbahrii isolates were identified. Diagnostic assays for P. belbahrii isolates were developed and validated in the lab, field, and using seed samples. Co-PI Gent supported diagnostic assays for air sampling. Co-PI Ojiambo conducted field experiments to assess the survival of P. cubensis as oospores. Oospores survived during the winter and were still viable at the start of the planting season. Co-PI Quesada determined that weed and wild cucurbit hosts can serve as reservoirs for P. cubensis. Follow up work not only confirmed the host adaptation of clade 1 and clade 2 isolates in North Carolina; clade 2 isolates arrive earlier in NC. PD Hausbeck refined techniques for sampling and extracting DNA to create a genotypic library to compare P. cubensis (clades 1 and 2 from different locations, hosts and years). The results showed little genetic variation in P. cubensis from locations around Michigan, but significantly larger differences between Michigan and samples from southern states. This supports the theory that P. cubensis may be overwintering in Michigan greenhouses, then spreading to fields. Co-PIs Bhattacharyya, Gent, and Ojiambo developed spatio-temporal models to account for multiple sources of biological species that can govern disease gradients and spatial spread in time. This modeling strategy can be used to predict DM spread, using only first incidence and wind data. The effect of early season disease prevalence on the model indicate multiple disease sources. The model proved tractable for estimating the generalized location and velocity of a disease front from sparsely sampled data with minimal data acquisition costs. PD Hausbeck and co-PIs Smart, Vallad, and Wyenandt conducted fungicide efficacy trials each year; cucurbit DM trials in MI, NY, and FL and a basil DM trial in NJ. Treatments evaluated included conventional and organic fungicides, biopesticides, and application timing schedules. Objective 3. Co-PIs Mazourek, Mou, and Simon bred cucumber, spinach, and basil plant populations with increased DM-resistance and good horticultural traits. PD Hausbeck and Co-PIs Quesada, Agehara, Mazourek, and Smart collaborated on a cucumber participatory breeding project and field trials of new cultivars. Co-PI Quesada collaborated with seed companies to test CDM-resistant cucumber lines, supporting the release of two varieties. Co-PI Mazourek selected and released nine new cucumber breeding lines to the seed industry. Seven of these are pickling types, developed by crosses between our best sources of DM resistance with grower preferred, cucumbers that lacked intellectual property restrictions. Also two new DM resistant slicing cucumbers with improved shape and earliness were developed and released. Co-PI Simon collected 500 basil accessions from around the world and evaluated the commercial lines. Some varieties marketed as DMR were not and several newer varieties of basil DMR that exhibited different levels of resistance, were identified. New Thai basils (20) with high DMR and aroma were created. Four DMR sweet basils were patented. One lemon basil family with DMR-resistance was identified. Co-PI Mou conducted RNA-seq analyses of transcriptomic changes in spinach cultivars to assess candidate genes necessary for DM resistance in spinach. The results supported the hypothesis that oospores imported on seeds, serve as primary inoculum sources for spinach DM. Recurrent selection breeding led to DM-resistant spinach and biopesticides tested in field trials also minimized DM. Objective 4. A survey developed by Co-PI Govindasamy showed that although more a virulent strain of CDM is present, cucurbit crop losses were significantly reduced in the last cropping cycle, without use of additional fungicides. This may be attributed to grower education about efficacious fungicide programs. In the initial survey, about 50% of the farmers were using more fungicides than 5 years ago. Another survey indicated that over 75% of basil growers in Florida are now using DM-resistant lines. Information developed and disseminated during this project also increased grower profits. Objective 5. Yearly sentinel plots were established; Co-PI Ojiambo facilitated CDMipmPIPE and Basil Ag Pest Monitor for reporting outbreaks by PD and Co-PIs. Workshops, field days, trial tours, open houses, extension articles, websites, fact sheets, social media, grower and professional meetings, and coordinated outreach with extension agents kept DM stakeholders updated on disease outbreaks and management. Eight DM-resistant sweet basil cultivars (patent #10159212) by Co-PIs Simon and Wyenandt and tested by PD Hausbeck and Co-PIs Quesada, Raid, Smart, and Vallad, became available. Over 75% of basil in FL are DM resistant. Field losses due to lettuce DM have been negligible since 2018, and basil acreage has rebounded from pre-2016 levels with use of resistant cultivars. Objective 6. Participation of underrepresented students in agriculture was increased and students were mentored for graduate school or agricultural careers. Students received their PhDs (3) and MS (1) with the PD/PIs. Post-doctoral researchers (7), graduate (25), and undergraduate (68) students were mentored. Courses (2) were developed and one enhanced by Co-PI Miles (over 150 undergrads) and shared with non-project professors. Undergraduate summer research included 37 students. Students (33) were accepted at NSF Research Experiences for Undergraduates (REU) at 5 eastern US sites. Awards were won by students (NCSU Kelman Scholars Awards, SACNAS poster, MSU Plant Genomics REU, and Outstanding Future Alumni and Research Fellowship).

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Adams, M. L., DArcangelo, K. N., Quesada-Ocampo, L. M. 2020. Evaluation of fungicides and cultivars for control of cucumber downy mildew. Phytopathology 110: S1.21.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: DArcangelo, K. N., Rahman, A., Miles, T. D. and Quesada-Ocampo, L. M. 2021. Utilizing a population genetics approach to facilitate crop-specific management of the cucurbit downy mildew pathogen, Pseudoperonospora cubensis. Phytopathology (in press).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: DArcangelo, K. N., Rahman, A., Miles, T. D. and Quesada-Ocampo, L. M. 2020. Leveraging population genetics to develop disease control practices: a study in the crop-specific management of cucurbit downy mildew. Phytopathology 110: S2.203.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: DArcangelo, K. N., Rahman, A., Miles, T. D., and Quesada-Ocampo, L. M. 2020. Utilizing a population genetics approach to provide crop-specific management strategies for cucurbit downy mildew. Phytopathology 110: S1.7.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Hausbeck, M.K. 2020. Downy Mildew Management in Pickling Cucumbers. Great Lakes Farm, Fruit and Vegetable Expo. Virtual, 8 Dec. Pickling Cucumber 1 Session Summaries.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Hausbeck, M.K. and Higgins, D.S. 2020. The Grounder, the Line Drive, and the Pop Fly: Fielding Three Very Different Vine Crop Diseases. Great Lakes Farm, Fruit and Vegetable Expo. Virtual, 9 Dec. Vine Crops Session Summaries.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Johnson E., Tian M., and Quesada-Ocampo, L. M. 2020. Differential expression of genes encoding sugar transporters in the basil pathogen Peronospora belbahrii. Phytopathology 110: S2.72.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Purayannur, S., Cano, L. M., Bowman, M. J., Childs, K. L., and Quesada-Ocampo, L. M. 2020. Clade-specific RXLR effectorome of the cucurbit downy mildew pathogen Pseudoperonospora cubensis. Phytopathology 110: S2.6.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Purayannur, S., Cano, L.M., Bowman, M., Childs, K.L., and Quesada, L.M. 2020. Host-specific effectors of the cucurbit downy mildew pathogen Pseudoperonospora cubensis. Phytopathology 110: S1.3.
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Raid, R. N., L. Rodrigues, E. Cooper, P. Watanabe, A. Hartman, L. Lopez, and G. Sandoya. 2021. Evaluation of Systemic Acquired Resistance (SAR) inducers for management of downy mildew (Hyaloperonospora parasitica) on baby leaf kale (Brassica oleracea Lacinata). Proc. S. Div. Assoc. of Plant Biologist (in press).
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Raid, R. N., L. Rodrigues, E. Cooper, P. Watanabe, A. Hartman, L. Lopez, and G. Sandoya. 2021. Evaluation of various rates of potassium phosphite for management of downy mildew on baby leaf kale. Proc. S. Div. Assoc. of Plant Biologist (in press).
  • Type: Conference Papers and Presentations Status: Awaiting Publication Year Published: 2021 Citation: Standish, J. R., Bowman, M. J., Childs, K. L., Tian, M., and Quesada-Ocampo, L. M. 2021. Development and validation of Peronospora belbahrii-specific diagnostic markers. Phytopathology: in press.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Standish, J. R., Bowman, M., Childs, K. L., Tian, M., and Quesada-Ocampo, L. M. 2020. Utilizing comparative genomics to develop species-specific diagnostic markers for basil downy mildew. Phytopathology 110: S2.59.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Standish, J. R., Purayannur, S., Bowman, M. J., Childs, K. L., Tian, M., and Quesada-Ocampo, L. M. 2020. Predicting the Peronospora belbahrii secretome for in silico identification of effector proteins. Phytopathology 110: S1.18.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Adams, M. L., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of fungicides for control of downy mildew on cucumber, Clayton, 2019. Plant Disease Management Reports 14:V116.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Adams, M. L., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of cultivars in combination with fungicides for control of downy mildew and yield effects on cucumber, Clinton, 2019. Plant Disease Management Reports 14:V117.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Adams, M. L., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of fungicides and cultivars for control of downy mildew on cucumber, Kinston 2019. Plant Disease Management Reports 14:V107.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Adams, M. L., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of fungicides for control of downy mildew on cucumber, Kinston II, 2019. Plant Disease Management Reports 14: V106.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Adams, M. L., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of fungicides for control of downy mildew on cucumber, Kinston III, 2019. Plant Disease Management Reports 14: V105.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: Bello Rodriguez, J.C., Sakalidis, M.L., Perla, D., and Hausbeck, M.K. 2021. Detection of airborne sporangia of Pseudoperonospora cubensis and P. humuli in Michigan using Burkard spore traps couple to qPCR. Plant Disease (first look). https://doi.org/10.1094/PDIS-07-20-1534-RE
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Bello, J. C., Engfehr, C.L. and Hausbeck, M.K. 2020. Evaluation of alternating programs of fungicides for control of downy mildew on pickling cucumber, 2019. Plant Disease Management Reports 14:V163.
  • Type: Journal Articles Status: Accepted Year Published: 2021 Citation: Bello, J.C., Hausbeck, M.K., and Sakalidis, M.L. 2021. Genotyping of the obligate plant pathogens Pseudoperonospora cubensis and P. humuli using target enrichment sequencing. Molecular Plant-Microbiome Interactions (accepted with revisions).
  • Type: Journal Articles Status: Submitted Year Published: 2021 Citation: Bello, J.C., Sakalidis, M.L., Quesada-Ocampo, L.M., Martin, F., and Bello, J.C., Sakalidis, M.L., Quesada-Ocampo, L.M., Martin, F., and Hausbeck, M.K. 2021. Advancing the monitoring of airborne Pseudoperonospora spp. sporangia using spore traps and qPCR for disease management of cucurbit downy mildew. Phytopathology (submitted).
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Bhattarai, G., A. Shi, C. Feng, B. Dhillon, B. Mou, and J. C. Correll. 2020. Genome wide association studies in multiple spinach breeding populations refine downy mildew Race 13 resistance genes. Frontiers in Plant Science 11:563187. https://doi.org/10.3389/fpls.2020.563187
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Clark, K.J., C. Feng, B. Dhillon, S.L. Kandel, B. Poudel, B. Mou, S.J. Klosterman, and J.C. Correll. 2020. Evaluation of spinach cultivars for downy mildew resistance in Yuma, AZ 2020. Plant Disease Management Reports 14: V146.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: Crandall, S. G., Ramon, M. L., Burkhardt, A. K., Bello, J. C., Adair, N., Gent, D. H., Hausbeck, M. K., Quesada-Ocampo, L. M., and Martin, F. N. 2021. A multiplex TaqMan qPCR assay for detection and quantification of clade 1and clade 2 isolates of Pseudoperonospora cubensis and P. humuli. Plant Disease (first look). https://doi.org/10.1094/PDIS-11-20-2339-RE
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: DArcangelo K. N., Adams M. L., Kerns J. P., and Quesada-Ocampo L. M. 2021. Assessment of fungicide product applications and program approaches for control of downy mildew on pickling cucumber in North Carolina. Crop Protection 140: 105412.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: DArcangelo, K. N., Rahman, A., Miles, T. D., and Quesada-Ocampo, L. M. 2020. Utilizing a population genetics approach to provide crop-specific management strategies for cucurbit downy mildew. Phytopathology 110: S1.7.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: Govindasamy, R., Hausbeck, M.K., Simon, J., and Wyenandt, A. 2021. Downy Mildew Impacts and Control Measure on Cucurbits in the United States. Journal of the American Society of Farm Managers and Rural Appraisers (in press).
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: Govindasamy, R., S. Arumugam, G. Gao, M. Hausbeck, A. Wyenandt and J.E. Simon. Downy Mildew Impacts and Control Measures on Cucurbits in the United States. Journal of American Society Of Farm Managers And Rural Appraisers 2021. (In Press).
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Govindasamy, R., S. Arumugam, G. Gao, M. Hausbeck, A. Wyenandt and J.E. Simon. 2020. Cucurbit Downy Mildew in High Value Crops: A Producer Survey. Journal of New Jersey Agricultural Experiment Station Report, December 2020.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Hatlen, R.J., Gillett, J.M., Sysak, R.W., Smith, R.L., Miles, T.D. 2020. Evaluation of fungicides for control of downy mildew in hops, 2019. Plant Disease Management Reports 15:OT018.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: Higgins, D., Miles, T.D., Hausbeck, M. 2021. Fungicide efficacy against Pseudoperonospora humuli and assessment of point-mutations linked to carboxylic acid amide (CAA) resistance in Michigan. Plant Disease (First Look). https://doi.org/10.1094/PDIS-01-20-0023-RE
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Kandel, S., A. Hulse-Kemp, K. Stoffel, S. Koike, A. Shi, B. Mou, A. van Deynze, and S. Klosterman. 2020. Transcriptional analyses of differential cultivars during resistant and susceptible interactions with Peronospora effusa, the causal agent of spinach downy mildew. Scientific Reports 10: 6719. https://doi.org/10.1038/s41598-020-63668-3
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Kenny, G.E., Engfehr, C.L. and Hausbeck, M.K. 2020. Evaluation of single product treatments for control of downy mildew on pickling cucumbers, 2019. Plant Disease Management Reports 14:V183.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Ojiambo, P. S., Gent, D. H., Mehra, L. K., Christie, D., and Magarey, R. 2017. Focus expansion and stability of the spread parameter estimate of the power law model for dispersal gradients. PeerJ 5:e3465. https://doi.org/10.7717/peerj.3465.
  • Type: Journal Articles Status: Accepted Year Published: 2021 Citation: Ojwang, M., Ruiz, T., Bhattacharyya, S., Chatterjee, S., Ojiambo, P. S., and Gent, D. H. 2021. A general framework for spatio-temporal modeling of epidemics with multiple epicenters: Application to an aerially dispersed plant pathogen. PLoS Computational Biology. (Accepted pending revision).
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Purayannur, S., Gent, D.H., Miles, T.D., Radisek, S., Quesada-Ocampo, L.M. 2021. The hop downy mildew pathogen Pseudoperonospora humuli. Molecular Plant Pathology. Online. https://doi.org/10.1111/mpp.13063.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Purayannur, S., Miles, T.D., Gent, D.H., Pigg, S., Quesada-Ocampo, L.M. 2020. Hop downy mildew caused by Pseudoperonospora humuli: a diagnostic guide. Plant Health Progress 21:173179. https://doi.org/10.1094/PHP-10-19-0072-DG.
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Rahman, A., Standish J.R., DArcangelo K. N., and Quesada-Ocampo L. M. 2021. Clade-specific biosurveillance of Pseudoperonospora cubensis using spore traps for precision disease management of cucurbit downy mildew. Phytopathology 111: 312-320.
  • Type: Journal Articles Status: Submitted Year Published: 2021 Citation: Raid, R. N., A. Hartman, L.Rodrigues, and G. Sandoya. 2021. Influence of oxathiopiprolin rate on downy mildew incidence and severity in lettuce in Florida. Plant Disease Management Reports 15: (submitted).
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2021 Citation: DArcangelo, K. N., Rahman, A., Miles, T. D. and Quesada-Ocampo, L. M. 2021. Utilizing a population genetics approach to facilitate crop-specific management of the cucurbit downy mildew pathogen, Pseudoperonospora cubensis. Phytopathology: in press.
  • Type: Journal Articles Status: Submitted Year Published: 2021 Citation: Raid, R. N., L. Rodrigues, E. Cooper, P. Watanabe, A. Hartman, L. Lopez, and G. Sandoya. 2021. Evaluation of Systemic Acquired Resistance (SAR) inducers for management of downy mildew (Hyaloperonospora parasitica) on baby leaf kale (Brassica oleracea Lacinata). Plant Disease Management Reports 15: (submitted).
  • Type: Journal Articles Status: Submitted Year Published: 2021 Citation: Raid, R. N., L. Rodrigues, E. Cooper, P. Watanabe, A. Hartman, L. Lopez, and G. Sandoya. 2021. Evaluation of various rates of potassium phosphite for management of downy mildew on baby leaf kale. Plant Disease Management Reports 15: (submitted).
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Salcedo A., Purayannur S., Standish J. R., Miles T., Thiessen L., and Quesada-Ocampo L. M. 2021. Fantastic downy mildew pathogens and how to find them: Advances in detection and diagnostics. Plants 10: 435.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Salcedo, A., Hausbeck M. K., Pigg S., and Quesada-Ocampo L. M. 2020. Diagnostic guide for cucurbit downy mildew. Plant Health Progress 21: 166-172.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Salcedo, A., Hausbeck, M., Pigg, S., and Quesada-Ocampo, L.M. 2020. Diagnostic guide for cucurbit downy mildew. Plant Health Progress 21:166-172. https://doi.org/10.1094/PHP-12-19-0095-DG
  • Type: Journal Articles Status: Submitted Year Published: 2021 Citation: Shirley, A.M., G.E. Vallad, N. Dufault, R. Raid, and L.M. Quesada-Ocampo. 2021. Duration of disease control for fungicides against cucurbit downy mildew under Florida field conditions. Plant Disease (submitted).
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Shirley, A.M., G.E. Vallad, S. Kalb, B.S. Hughes, and R. Willis. 2019. Evaluation of selected fungicides for management of downy mildew on cucumber, fall 2018. Plant Disease Management Reports 14: V085.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Shirley, A.M., G.E. Vallad, S. Kalb, B.S. Hughes, and R. Willis. 2017. Evaluation of selected fungicides for management of downy mildew on cucumber, fall 2016. Plant Disease Management Reports 11: V060.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish J. R., Collins H., and Quesada-Ocampo L. M. 2020. Evaluation of fungicides for managing basil downy mildew epidemics in North Carolina, 2019. Plant Disease Management Reports 14: V032.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish, J. R. and Quesada-Ocampo L. M. 2020. Evaluation of sweet basil cultivars for downy mildew tolerance in North Carolina, 2020. Plant Disease Management Reports 15: V059.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish, J. R., Adams M. L., and Quesada-Ocampo L. M. 2020. Evaluating basil cultivars for tolerance to downy mildew in North Carolina, 2019. Plant Disease Management Reports 14: V033.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish, J. R., Collins H., and Quesada-Ocampo L. M. 2020. Evaluating fungicides to manage epidemics of basil downy mildew in North Carolina, 2020. Plant Disease Management Reports 15: V058.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish, J. R., Raid R. N., Pigg S., and Quesada-Ocampo L. M. 2020. A diagnostic guide for basil downy mildew. Plant Health Progress 21: 77-81.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Standish, J. R., Sharpe S., Butler S., Quesada-Ocampo L. M., and Meadows I. 2020. First report of downy mildew, caused by Peronospora effusa, on spinach (Spinacea oleracea L.) in North Carolina. Plant Health Progress 21: 194-196.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wallace, E., DArcangelo K. N., and Quesada-Ocampo L. M. 2020. Population analyses reveal two host-adapted clades of Pseudoperonospora cubensis, the causal agent of cucurbit downy mildew, on commercial and wild cucurbits. Phytopathology 110: 15-78-1587.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wyenandt, C.A. 2020. Evaluation of fungicides for the management of downy mildew in cucumber, 2016. Plant Disease Management Reports 14:V171.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wyenandt, C.A. 2020. Evaluation of fungicides for the management of downy mildew in cucumber, 2017. Plant Disease Management Reports 14:V172.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wyenandt, C.A., and J.E. Simon. 2020. Evaluation of biopesticide and conventional fungicides for management of downy mildew in sweet basil, 2019. Plant Disease Management Reports. 14:V175.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Wyenandt, C.A., and J.E. Simon. 2020. Evaluation of fungicide programs for management of downy mildew in sweet basil, 2017. Plant Disease Management Reports. 14:V174.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M. 2020. Cucumber downy mildew moves across Michigan. MSU Extension News: Vegetables:30 Jun. https://www.canr.msu.edu/news/cucumber-downy-mildew-moves-across-michigan.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M. 2020. Cucumber downy mildew update on spore detection. MSU Extension News: Vegetables:17 Jun. https://www.canr.msu.edu/news/cucumber-downy-mildew-update-on-spore-detection.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M. 2020. First cucumber downy mildew outbreak of 2020 confirmed in Berrien County. MSU Extension News: Vegetables:22 Jun. https://www.canr.msu.edu/news/cucumber-downy-mildew-outbreak-of-2020.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M.K. 2020. Downy Mildew Team Develops a New Tool to Detect Cucurbit Pathogen in Air Samples. Vegetable Growers News: Oct 2020.
  • Type: Other Status: Published Year Published: 2020 Citation: Tekip, A. 2020. Monitoring and managing cucumber downy mildew. Michigan State University AgBioResearch, Futures: Summer 2020. https://www.canr.msu.edu/news/monitoring-and-managing-cucumber-downy-mildew.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M.K., Linderman, S.D., and Higgins, D.S. 2020. Managing cucurbit downy mildew. Fact Sheet. https://veggies.msu.edu/extension-publications/#FactSheets.
  • Type: Other Status: Published Year Published: 2020 Citation: Hausbeck, M.K., Linderman, S.D., and Higgins, D.S. 2020. Monitoring cucurbit downy mildew. Fact Sheet. https://veggies.msu.edu/extension-publications/#FactSheets.
  • Type: Other Status: Published Year Published: 2020 Citation: Quesada-Ocampo L. M. and DArcangelo, K. N. Pumpkin downy mildew found in Surry county, North Carolina. Extension Plant Pathology Portal. August 21, 2020.
  • Type: Other Status: Other Year Published: 2020 Citation: Quesada-Ocampo L. M. and Parker B. Cucumber downy mildew found in Johnston County. Extension Plant Pathology Portal. June 18, 2020.
  • Type: Other Status: Published Year Published: 2020 Citation: Quesada-Ocampo L. M., DArcangelo, K. N, Adams M., and Batts T. Cucumber downy mildew confirmed in North Carolina. Extension Plant Pathology Portal. June 8, 2020.
  • Type: Theses/Dissertations Status: Awaiting Publication Year Published: 2021 Citation: Bello, J.C. 2020. Genetic Monitoring of Cucurbit Downy Mildew in Michigan. Dissertation. Michigan State University, East Lansing, MI.


Progress 03/01/19 to 02/29/20

Outputs
Target Audience:Our target audience includes people in the scientific, extension, and agricultural communities. Included are other scientists, undergraduate and graduate students, and post-docs in plant sciences, plant breeding and plant pathology, crop consultants, extension agents, crop protection industry, growers, shippers, processors, and other allied stakeholder industries. Changes/Problems:Due to the COVID-19 pandemic, co-PI Smart will not be able to conduct field trials or host an undergraduate in 2020. Other project field trials and lab work may be amended or shortened due to the limitations COVID-19 safety restrictions on travel to field sites and staggering personnel in the lab. These changes are not expected to impede completing work on the main objectives of this project. What opportunities for training and professional development has the project provided?This project has resulted in the training of many undergraduates, graduate students, and post-doctoral scientists in both field- and laboratory-based agricultural scientific techniques. Field-based techniques include conventional and q-PCR, Burkard volumetric spore trapping and rotorod spore trapping, weather monitoring, establishing and evaluating greenhouse and field fungicide/biopesticide efficacy trials, sentinel plots, and cultivar trials including cucurbit, lettuce, spinach, and basil production, field plot design, establishment, seed germination, transplanting, maintenance, fungicide application, disease diagnostics/PCR diagnostics, disease evaluation, yield assessments, canopy cover imaging and analysis, greenhouse plant production, cucumber pollinations and seed processing, cross and self-pollination and drying of basil, organizing breeding populations in the field, and prioritizing selections. Laboratory-based techniques include DM pathogen sampling and sporangial and DNA preservation, transfers of DM onto new host plant material and cell counting, growth and storage of P. cubensis and P. humuli, DNA extraction, PCR, qPCR, SSR/microsatellite genotyping sequencing analyses and Sanger sequencing, design and development of diagnostic assays including the mitochondrial-based TaqMan Diagnostic assay. Other techniques learned included statistics, network modeling approaches in biological systems, image analysis techniques for disease damage quantification, biomathematics on network modeling of cucurbit DM, genomics, bioinformatics, epidemiology, the role of oospores in the pathogen survival and epidemiology, designing, developing and testing of the website for tracking basil DM, coordination and compilation of grower survey results, and preparation of presentations, technical reports, and manuscripts. Post-doctoral scientists, graduate students, and undergraduate students have been trained in the methods and techniques listed above as required for this project under the supervision of the PD and co-PIs and aided by technicians. Those who have benefited from this training include post-doctoral scientists (Safa Alzohairy, Elizabeth Brisco, Alyssa Burkhardt, Shyam Kandel, Alamgir Rahman), graduate students (Julian Bello, Lara Brindisi, Kim D'Arcangelo, Doug Higgis, Sara Getson, Juan Gonzalez Giron, Grace Kenny, Robert Mattera, Anthony Noto, Maureiq Ojwang, David Perla, Robert Pyne, Trevor Ruiz, Andy Shirley, Martha Sudermann, Tianxin Wu), undergraduate students (Andew Abdelmalek, Carson Adams, Stephanie Aponte, Tanner Boss, Baylee Carr, Jordan Castellari, Jia-Yi Chan, Brian Change, Maeve Day, Abby Dooley, Paul Anthony Edwards, Morgan Gifford, Amber Glenn, Ryan Gould, Asia Green, William Gura, Ross Hatlen, Lexi Heger, Emily Henderson, Madeline Hendrickson, Clayton Jarvis, Adam Job, Kavindi Karunaratne, Nicholas King, Elizabeth Kogan, Krupali Kothari, Jacob Kowaleski, Courtney Lepping, Vicki Meraz, Laura Merja, Kelly Merrill, Sara Merrill, Reilly Nakomoto, Han Nguyen, A.J. Noto, Lisa Panichelli, Benjamin Rouse, Carly Seldow, Trevor Styles, Halley Taddonio, Opal Tam, Jessica Tan, Jonathan Vantman, Yazmin Williams, Danielle Wolleman), and technicians (Tanner Boss, Alex Cook, Uma Crouch, Colin Day, Christopher DelCastillo, Allison DeSario, Cheryl Engfehr, Jack Fabrizio, Garrett Giles, Asia Green, Blair Harlan, Taylere Herrmann, Barry Hughes, Scott Hughes, Gregory Inzinna, Marc Jaquez, John Kurtz, Holly Lange, Sheila Linderman, Marcus Lopez, Patricia Love, Leah Martin, Mike Matson, Marina Ramon, Paige Reeves, Emily Rustad, Joe Shemanski, Sheri Smith, Shane Sullivan, Richard Zimmerman). How have the results been disseminated to communities of interest?Research results have been disseminated via grower demonstration plots, one-on-one grower conversations and farm visits, publications, websites, scientific/extension presentations, webinars, and scientific/extension meetings. NIFA was acknowledged in 16 presentations. Scientific Meetings: Results were disseminated to the scientific community in 9 presentations at scientific meetings, including the Annual Meeting of the American Phytopathological Society, Cleveland, OH (7 presentations) and the Plant Science Symposium, Rutgers, the State University of New Jersey, New Brunswick, NJ (2 presentations). Extension Meetings: Research results were disseminated as extension presentations at 36 meetings at the local, regional, and national levels, to growers, commodity groups, chemical companies, extension educators, and other agricultural industry stakeholders. Meetings included including the USDA NIFA Basil Workshop, Everglades Research and Education Center, University of Florida, Belle Glade, FL, 15 Dec 2019 (7 presentations), three Phytophthora and Cucurbit Downy Mildew Workshops, Benton Harbor, MI, 23 Jan 2019 (1 presentation), Frankenmuth, MI, Feb 2019 (4 presentations), Hart, MI, Mar 2019 (2 presentations), Pickle Packers International Annual Meeting, St. Petersburg, FL, Oct 2019 (2 presentations), Pickle Packers International Spring Meeting, Detroit, MI, Apr 2019, Great Lakes Farm, Fruit and Vegetable Expo, Grand Rapids, MI, Dec 2019, Growers' Meeting, Davison, MI, 6 Mar, Southwest Michigan Winter Extension Meeting, Benton Harbor, MI, 2019, Clifford E. & Melda C. Snyder Research and Extension Farm, Pittstown, NJ, Aug 2019, NJ-NOFA Annual Conference, New Brunswick, NJ, 2019, California Leafy Greens Research Program (CLGRP) Annual Meeting, Pismo Beach, CA, 19 March 2019, California Leafy Greens Research Program Mid-year Meeting, Salinas, CA, 8 Oct 2019, Eastern New York Fruit & Vegetable Conference, Albany, NY, 26 Feb 2020, Western New York Vegetable Growers Meeting (Zoom), 20 Feb 2020, Long Island Agricultural Forum Riverhead, NY, 9 Jan 2020, Empire Farm Days, Seneca Falls, NY, 8 Aug 2019, Eastern New York Winter Vegetable Meeting, Albany, NY, 20 Feb 2019, Empire State Producers Expo, Syracuse, NY, 15 Jan 2019, Suwannee Valley Watermelon Institute, Gainesville, FL, Dec 2019, Syngenta Annual Research Meeting, Vero Beach, FL, 2019, Citrus Expo and Vegetable & Specialty Crop Expo, Fort Myers, FL, Aug 2019, North Carolina Vegetable Growers Association Ag Expo, Wilmington, NC, Dec 2019, Agent Training on Disease Diagnostics and Management in Vegetable Crops, Clayton and Raleigh, NC, Jul 2019, and Small Farms Tour: Disease Diagnostics and Management in Vegetable Crops, Clayton, NC, Jun 2019. Webinars, open houses, workshops, field days, trial tours: Four workshops were scheduled to feature this project's research: USDA NIFA Basil Workshop (Belle Glade, FL, 15 Dec 2019), and three Phytophthora and Cucurbit Downy Mildew Workshops (Hart, MI, 5 Mar 2019, Frankenmuth, MI 14 Feb 2019, and Benton Harbor, MI, 24 Jan 2019). Four articles were authored by people other than project PD/co-PIs and featured project research were published in Edible Jersey, GrowerTalks, Greenhouse Grower, and Inside Grower. What do you plan to do during the next reporting period to accomplish the goals?The effects of the COVID-19 pandemic will impact research and field trials for the upcoming reporting period; PD Hausbeck and all co-PIs will endeavor to continue this project. Objective 1: Early detection and rapid diagnostics. Co-PI Martin will continue to develop mitochondrial diagnostic markers and provide sequencing support. Co-PI Quesada will continue to identify nuclear markers and provide sequencing support. PD Hausbeck and co-PIs Gent and Quesada will continue to test markers and provide isolates. Co-PIs Ojiambo, Raid, and Smith will continue to provide isolates. Co-PI Smart will test isolates collected in NY in 2020 using markers developed as part of this project, if protocols are available. Co-PI Miles will continue assay development and marker validation. Objective 2: Tracking pathogen dispersal and survival. Co-PI Ojiambo will continue ipmPIPE administration. PD Hausbeck and co-PIs Quesada, Smart, Smith, Vallad, and Wyenandt will continue sentinel plot deployment to track movement of DM disease on cucurbits and basil. PD Hausbeck will continue to monitor airborne sporangia and compare efficacy of molecular diagnostics via PCR rotorod spore traps versus conventional traps with markers supplied by co-PI Martin. Co-PI Smart will test from leaf samples, not spore traps in 2020. Co-PI Wyenandt will test seeds with molecular diagnostics developed by this project. Co-PIs Ojiambo, Bhattacharyya, and Gent will continue developing modeling to provide grower alerts based on inoculum source, strength, and wind patterns; PD Hausbeck and co-PIs Ojiambo, Quesada, Smart, Vallad, and Wyenandt will contribute to provide data for the national tracking website. PD Hausbeck and co-PIs Vallad and Wyenandt will continue to monitor duration of DM control with fungicide efficacy trials. Co-PI Smart will not have a DM fungicide trial in 2020 due to the COVID-19 shutdown. Objective 3: Developing dm-resistant crops. Co-PI Mou will plant seeds harvested from 34 populations with resistant and susceptible controls. The field plots will be kept wet with sprinkler irrigation to promote downy mildew development and sporulation. Leaf samples with spores of the pathogen will be collected and sent to the plant pathology lab at University of Arkansas to identify pathogen races using differential hosts. Downy mildew disease incidence of each breeding population as well as resistant/susceptible controls will be recorded. Resistant plants will be selected again from each population and transplanted into isolators for seed production. This cycle of recurrent selection will continue until a satisfactory level of resistance is achieved. Then the populations will be tested in the field for downy mildew resistance, uniformity, horticultural traits, and yield. Co-PI Simon will continue to evaluate new basil seed for DM and newly available DM-resistant basil cultivars for field resistance to DM. New crosses and selections of basil will continue to be evaluated for field performance and DM resistance. Basil DNA will continue to be extracted and all lines will be sequenced and genotyped with SNPs and SSR markers. Co-PI Mazourek will sow seeds from plants produced by cuttings from 13 lines selected and evaluated in 2019 in a replicated complete block design and will evaluate for fruit number, shape, seed cavity, and disease resistance. PD Hausbeck and co-PI Agehara will establish participatory field trials in MI and FL to evaluate/select the breeding lines with commercial growers and extension agents. Objective 4: Cost/benefit ratios and grower adoption. PD and co-PIs will continue to administer surveys prepared by co-PI Govindasamy and collect data as needed to evaluate costs/benefits and grower adoption. Objective 5: Outreach and adoption. Co-PI Ojiambo will continue to coordinate monitoring and reporting of disease outbreaks using sentinel plots and uploaded disease forecasts and disease alerts to DM stakeholders during the growing season. PD Hausbeck and co-PIs Agehara, Quesada, Raid, Smart, Smith, and Vallad will continue to report DM outbreaks to the ipmPIPE monitoring/forecasting system and train growers and extension personnel in the use of this system. Grower outreach educational programs will be planned for the 2020 growing season. PD Hausbeck and co-PIs Quesada, Raid, Vallad, and Wyenandt will test breeding lines/varieties in their regions for DM resistance as needed. Because of shut down due to COVID-19, co-PI Smart will not be testing DM resistant breeding lines in 2020. PD Hausbeck and co-PIs Agehara and Smith will evaluate breeding lines/varieties in their region for horticultural traits. PD Hausbeck and co-PIs will conduct basil seed testing experiments, including testing commercial seed for the presence of DM and continue to develop/disseminate educational materials on the importance of clean seed. PD Hausbeck and co-PIs Quesada, Raid, Smart, and Wyenandt will continue developing and disseminating field spray recommendations based on regional trials. PD Hausbeck and co-PIs Agehara, Goldy, Quesada, Raid, Smart, Smith, Vallad, and Wyenandt will continue developing social media and posting frequent updates to advise growers. Objective 6: Attract students to agricultural sciences. Due to limited remaining funds, 1-2 additional students are expected to participate in a Research Experiences for Undergraduates program in the labs of co-PI Quesada at NCSU or Miles at MSU. Co-PI Smart will not host an undergrad during the summer of 2020 due to COVID-19. Co-PI Miles is currently conducting a survey of all undergraduate students impacted by this grant for reporting purposes.

Impacts
What was accomplished under these goals? Objective 1: Early detection and rapid diagnostics. Co-PI Martin used additional field samples to validate diagnostic markers and confirm relationship between spore counts and qPCR results and provided DNA samples to co-PIs Quesada, Smart and PD Hausbeck for calibrating qPCR results with sporangial counts. Co-PI Smart successfully compared assays from co-PIs Martin and Quesada. PD Hausbeck added an internal control to the qPCR assay that lowered quantification limit of the assay to 3 sporangia. PD Hausbeck used a combination of spore traps and the qPCR assay to rapidly detect and differentiate P. cubensis in the main cucumber growing regions of Michigan. Co-PI Miles designed several in silico assays to detect resistance to U15 fungicides. These markers still require sensitivity and specificity testing which would go along with assays developed for FRAC 11 and 40. Co-PI Miles developed a LAMP based assay capable of in-field detection for resistance of FRAC 11 fungicides. Objective 2: Tracking pathogen dispersal and survival. Sentinel plots for monitoring DM during the reporting year were planted in MI, NY, FL, NC, and NJ by PD Hausbeck and co-PIs Quesada, Smart, Vallad, and Wyenandt. A Burkard volumetric spore trap, ionic volumetric spore trap, and impact rod spore trap were all used at Florida sentinel plots by Co-PI Vallad. CDM outbreaks were reported from the west central FL sentinel plots and commercial cucurbit fields in nearby counties. PD Hausbeck collected and analyzed 1,000 samples and determined that P. humuli was more prevalent than P. cubensis in MI early in the growing season and P. cubensis clade II sporangia were detected 5-10 days before CDM symptoms were observed. The IPMpipe website continued to provide disease spread information throughout the season and based on grower surveys, >50% of cucurbit growers in NY are utilizing the alert system. Co-PIs Ojiambo, Bhattacharyya, and Gent used a new modeling strategy to predict DM spread, using only first incidence and wind data. The model assumed one inoculum source, however the effect of early season disease prevalence on the model pointed to multiple disease sources. The model worked well, showing the anisotropic model was more effective than an isotropic model. PD Hausbeck and co-PIs Smart, Vallad, and Wyenandt conducted fungicide efficacy trials; cucurbit DM trials in MI, NY, and FL and a basil DM trial in NJ. Treatments evaluated included conventional and organic fungicides, and biopesticides. Objective 3: Developing DM-resistant crops. Co-PI Simon compared 4 new downy mildew resistant lines, developed at Rutgers, with 17 commercially available cultivars, including varieties with varying claims of DMR. The Rutgers DMR lines performed well with generally high levels of resistance, but some exhibited higher levels of undesirable leaf appearance characteristics. Co-PI Simon continued to add new basil accessions to the Rutgers collection and screen for DMR to select the lines to use in breeding programs. Co-PI Mou used 34 spinach varieties that had been cross-bred from DMR cultivars to conduct a field trial in CA. Results of the field trial showed the breeding program was very effective at increasing downy mildew resistance. Resistant plants from this trial were transplanted into isolators for open-pollination and seed production. Co-PI Mou used RNA-seq to identify transcriptomic changes in spinach leaf tissue during different stages of DM infection. Objective 4: Cost/benefit ratios and grower adoption. Co-PI Vallad conducted field trials to assess the impact of CDM on cucumber yield and economic return of using a CDM fungicide program. Responses to Co-PI Govindasamy's survey from 98 cucurbit growers showed that 56% of farmers surveyed had a DM problem and 71% used a fungicide program, despite the added cost. Dollar loss due to DM ranged from $50-$1,425 per acre and yield loss was 137-1,372 pounds per acre. Overall, the farmers were concerned about yield loss and rising fungicide price and use. Most of the responding producers (67%) had not seen evidence of pathogen resistance to fungicides in the last year, but more than 50% felt they were using more fungicides than 5 years ago. Using plant host resistance as a preventative measure was noted by fewer than half of the respondents. Only about 50% of the farmers received disease management training from agricultural extension services, although the participants reported extension agents as their most relevant and preferred source of information. Objective 5: Outreach and adoption. Sentinel plots were established in the eastern U.S. and scouted weekly for disease. Regional reports of cucurbit DM occurrences to the cucurbit ipmPIPE website were kept up to date by the PD and co-PIs, ensuring accurate information was available to growers during 2019. Co-PI Ojiambo continued to coordinate the monitoring and reporting of disease outbreaks using sentinel plots and uploaded disease forecasts and disease alerts to DM stakeholders during the growing season. PD Hausbeck and Co-PI Quesada held open houses, field days, trial tours, workshops, and grower and professional meetings in 2019 to inform growers, extension agents, and industry personnel of 2019 research results and train growers how to identify and manage CDM through fungicides and other disease management practices. Fungicide efficacy and timing programs trials provided growers recommendations for managing DM that are up to date and region-specific. Cultivar trials were conducted; PD Hausbeck and Co-PI Quesada ran basil trials and Co-PI Smith ran a cucumber trial. Co-PI Raid surveyed Florida growers on use of novel DM resistant lines deployed in 2018 and University of Florida-recommended fungicide programs. 100% of survey respondents used both the recommended fungicide programs and tried the new lines with resulting increased production. Growers appreciated the disease resistance of the new basil lines, although plant horticultural trait reviews were mixed. Co-PI Wyenandt collected basil seed from DM resistant and susceptible lines to use for RT-PCR testing for BDM infestation. Trial results and disease updates were communicated quickly via lab websites and social media. Objective 6: Attract students to agricultural sciences. Andrew Shirley completed his Ph.D. (Thesis: Chemical control and population dynamics of Pseudoperonospora cubensis, the cause of cucurbit downy mildew in Florida) under the supervision of Co-PI Vallad. The PD and co-PIs have recruited 1 post-doc, 4 graduate students, and 14 undergraduate students to work on this project in the past year. This proposal has trained a number of undergraduate students and attracted them to plant sciences in general. Fourteen students have gone on to graduate school in plant science. At least 12 students also are currently working professionally in the agricultural industry in the Monterey Bay and Oklahoma areas at large agricultural companies. Course materials at CSUMB were enhanced by co-PI Miles for three courses (Plant Pathology, Plant Physiology and Systematic and Molecular Mycology) that have served over 150 undergraduates over the life of this project. Course materials have been shared with the new professor assuming leadership of this program to benefit future undergraduate students. Co-PI Miles transferred from CSUMB to MSU, modified a fungicide resistance laboratory to conduct downy mildew research and introduced a new lecture on biotrophic pathogens and symbiotic organisms. In the past year, three additional students attended a REU program at MSU or NCSU for a total of 33 undergraduate students partially or fully funded for research experiences in various laboratories or off-campus REU programs during this project. 100% of these students have graduated and 10 of them were accepted into graduate school in a Plant Science field (either MS or PhD).

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kandel, S., Mou, B., Shishkoff, N., Shi, A., Subbarao, K., and S. Klosterman. 2019. Spinach downy mildew: Advances in our understanding of the disease cycle and prospects for disease management. Plant Disease 103:791-803. https://doi.org/10.1094/PDIS-10-18-1720-FE (Cover story)
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Keinath, A.P., Miller, S.A., and Smart, C.D. 2019. Response of Pseudoperonospora cubensis to preventative fungicide applications varies by state and year. Plant Health Progress 20:142-146. https://doi.org/10.1094/PHP-04-19-0028-RS
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Mou, B. 2019. USDA Red spinach. HortScience 54:2070-2072. https://doi.org/10.21273/HORTSCI14308-19 (Cover story)
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides for control of downy mildew on hop, Clayton 2018. Plant Disease Management Reports 13:V066.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides for control of downy mildew on cucumber, Kinston 2018. Plant Disease Management Reports 13:V067.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides for control of downy mildew on cucumber, Kinston 2018. Plant Disease Management Reports 13:V068.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Goldenhar, K.E., and Hausbeck, M.K. 2019. Fungicides for control of downy mildew on pickling cucumber in Michigan. Plant Health Progress 20:165-169.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides for control of downy mildew on cucumber, Clinton 2018. Plant Disease Management Reports 13:V070.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides for control of downy mildew on cucumber, Clinton 2018. Plant Disease Management Reports 13:V071.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2019. Evaluation of cultivars in combination with fungicides for control of downy mildew and yield effects on cucumber, Clinton 2018. Plant Disease Management Reports 13:V072.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Adams, M.L., DArcangelo, K.N., Collins H., and Quesada-Ocampo, L.M. 2019. Evaluation of fungicides and cultivars for control of downy mildew on cucumber, Kinston 2018. Plant Disease Management Reports 13:V069.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: DArcangelo, K.N., Collins, H., Adams, M.L., and Quesada-Ocampo, L.M. 2019. Evaluation of cultivars for control of downy mildew on squash, Kinston 2018. Plant Disease Management Reports 13:V124.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: DArcangelo, K.N., Collins, H., Adams, M.L., and Quesada-Ocampo, L.M. 2019. Evaluation of cultivars for control of downy mildew on cucumber, Kinston 2018. Plant Disease Management Reports 13:V125.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Hausbeck, M.K., and Harlan, B.R. 2019. Evaluation of newly-released basil cultivars for resistance to downy mildew in the greenhouse, 2018. Plant Disease Management Reports 13:OT021.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Hausbeck, M.K., Perla, D.E., and Cook, A.J. 2019. Evaluation of single fungicide products for control of downy mildew of cucumber, 2018. Plant Disease Management Reports 13:V139.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Kandel, S.L., Subbarao, K.V., Shi. A., Mou, B., and Klosterman, S.J. 2019. Evaluation of biopesticides for managing downy mildew of spinach in organic production systems 2017 and 2018. Plant Disease Management Reports 13:V171.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Lange, H.W., Smart, C.D., and Seaman, A.J. 2019. Evaluation of materials allowed for organic production on downy mildew of cucumber, 2018. Plant Disease Management Reports 13:V096.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Pollok, J.R., Rideout, S.L., Wyenandt, C.A., Garcia-Gonzalez, J.F., Sutton, K.L., and Custis, J.T. Jr. 2020. Evaluation of basil cultivar resistance to downy mildew infection in Virginia, 2019. Plant Disease Management Reports 14:V036.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Raid, R.N., Hartman, A., and Reis, K. 2020. Evaluation of picarbutrazox for control of downy mildew on sweet basil. Plant Disease Management Reports 14:V039.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Raid, R.N., Hartman, A., and Sandoya, G. 2020. Evaluation of fungicides for control of downy mildew on crisphead lettuce. Plant Disease Management Reports 14:V036.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Raid, R.N., Hartman, A., and Sandoya, G. 2020. Evaluation of potassium phosphite alone and in combination with mancozeb for control of downy mildew on crisphead lettuce. Plant Disease Management Reports 14:V037.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Raid, R.N., Hartman, A., Reis, K., and Sandoya, G. 2020. Evaluation of picarbutrazox for control of downy mildew on crisphead lettuce. Plant Disease Management Reports 14:V038.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Perla, D.E., Cook, A.J., and Hausbeck, M.K. 2019. Evaluation of fungicide programs for control of downy mildew of cucumber, 2018. Plant Disease Management Reports 13:V147.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Downy mildew management in pickling cucumbers. Pages 2-3 in: Pickling Cucumber Session Summaries, Great Lakes Fruit, Vegetable and Farm Market Expo, Grand Rapids, MI, Dec. Online. https://glexpo.com/wp-content/uploads/2019/12/Pickling-Cucumber-Tuesday-AM.pdf
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Smart, C.D., and Lange, H. 2019. Disease update: What we saw in 2018 and what to expect in 2019. Proceedings of the Empire State Producers Expo, Jan.
  • Type: Theses/Dissertations Status: Published Year Published: 2020 Citation: Shirley, A. 2020. Chemical control and population dynamics of Pseudoperonospora cubensis, the cause of cucurbit downy mildew in Florida. Dissertation. University of Florida, Gainesville, FL.
  • Type: Other Status: Published Year Published: 2019 Citation: Bello, J., Hausbeck, M.K., and Sakalidis, M.L. 2019. Genotyping of the downy mildew species Pseudoperonospora cubensis and P. humuli using target genome sequencing. Abstract. Phytopathology 109(10S):S2.187.
  • Type: Other Status: Published Year Published: 2019 Citation: DArcangelo, K., and Quesada-Ocampo, L.M. 2019. Investigating the genetic structure of Pseudoperonospora cubensis populations after single-product fungicide application events. Abstract. Phytopathology 109:S2.167.
  • Type: Other Status: Published Year Published: 2019 Citation: Mandal, M.K., Ikerd, J., Wadl P.A., Williams, L., Quesada-Ocampo, L.M., and Kousik C.S. 2019. Comprehensive disease survey on USDA-ARS cucumber germplasm collections in search for downy mildew resistance at Charleston SC. Abstract. Phytopathology 109:S2.95.
  • Type: Other Status: Published Year Published: 2019 Citation: Quesada-Ocampo, L.M. 2019. Harnessing new technologies to improve management of cucurbit downy mildew. Abstract. Phytopathology 109:S2.198.
  • Type: Other Status: Published Year Published: 2019 Citation: Shirley, A., and Vallad, G.E. 2019. Developing cost-effective, sustainable fungicide programs for managing downy mildew on cucumber in Florida. Abstract. Phytopathology 109:S2.59.
  • Type: Other Status: Published Year Published: 2019 Citation: Shirley, A., Quesada-Ocampo, L.M., and Vallad. G.E. 2019. Effect of cucurbit host, production region, and season on the population structure of Pseudoperonospora cubensis in Florida. Abstract. Phytopathology 109:S2.169.
  • Type: Other Status: Published Year Published: 2019 Citation: Shirley, A., Quesada-Ocampo, L.M., and Vallad, G. 2019. Population genetic and structural analysis of Pseudoperonospora cubensis isolates in Florida. Abstract. Phytopathology 109:S2.169.
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Growers target downy mildew in cucumbers. MSUE Vegetable Notes. Vegetable Grower News: Jun, pp. 14-15.
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Cucumber downy mildew moves across Michigan: Apply fungicides proven to be effective immediately. Michigan State University Extension News-Vegetables:20 Aug. https://www.canr.msu.edu/news/cucumber-downy-mildew-moves-across-michigan
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Wet, humid weather challenges vegetable disease management. Michigan State University Extension News-Vegetables: 24 Jun. https://www.canr.msu.edu/news/wet-humid-weather-challenges-vegetable-disease-management
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M.K., Linderman, S.D., and Higgins, D.S. 2020. Managing cucurbit downy mildew. Online at https://veggies.msu.edu/extension-publications/#FactSheets
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Cucumber downy mildew disease confirmed in research plot in southwest Michigan. Michigan State University Extension News-Vegetables:29 Jul. https://www.canr.msu.edu/news/cucumber-downy-mildew-disease-confirmed-in-research-plot-in-southwest-michigan
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Cucumber downy mildew update. Michigan State University Extension News-Vegetables:22 Jul. https://www.canr.msu.edu/news/cucumber-downy-mildew-update
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M. 2019. Vegetable disease alert: Act now to protect cucumbers and onions. Michigan State University Extension News-Vegetables:8 Jul. https://www.canr.msu.edu/news/vegetable-disease-alert-act-now-to-protect-cucumbers-and-onions
  • Type: Other Status: Published Year Published: 2019 Citation: Hausbeck, M.K., Linderman, S.D., and Higgins, D.S. 2020. Monitoring cucurbit downy mildew. Online at https://veggies.msu.edu/extension-publications/#FactSheets
  • Type: Other Status: Published Year Published: 2019 Citation: Quesada-Ocampo, L.M., and DArcangelo, K. 2019. Cucumber downy mildew detected in North Carolina. Extension Plant Pathology Portal, 13 Jun. https://plantpathology.ces.ncsu.edu/2019/06/cucumber-downy-mildew-detected-in-north-carolina/
  • Type: Other Status: Published Year Published: 2020 Citation: Simon, J.E., Wyenandt, C.A., Raid, R.L., McGrath, M.T., and K. Homa. 2020. A plant breeding breakthrough: Downy mildew resistant sweet basil. Growing Produce, 29 Apr. https://www.growingproduce.com/vegetables/a-breeding-breakthrough-downy-mildew-resistant-sweet-basil/
  • Type: Other Status: Published Year Published: 2019 Citation: Vallad, G., and Shirley, A. 2019. Downy mildew of cucurbits. UF/IFAS GCREC Vegetable Pathology Program Fact Sheet. http://www.southeastagnet.com/documents/VegFactSheet-Cucurbits-downy%20mildew%20(2).pdf
  • Type: Other Status: Published Year Published: 2018 Citation: Vallad, G., and Shirley, A. 2018. Downy mildew of cucurbits. UF/IFAS GCREC Vegetable Pathology Program Fact Sheet.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Basil downy mildew confirmed in southern and central New Jersey  ALERT 6/30/16. https://plant-pest-advisory.rutgers.edu/basil-downy-mildew-confirmed-in-southern-and-central-new-jersey-alert-63016/
  • Type: Other Status: Published Year Published: 2019 Citation: Wyenandt, A. 2019. Basil downy mildew confirmed in southern New Jersey  ALERT 6/13/19. Plant & Pest Advisory, 13 Jun. https://plant-pest-advisory.rutgers.edu/basil-downy-mildew-confirmed-in-southern-and-central-new-jersey-alert-63016-2/
  • Type: Other Status: Published Year Published: 2019 Citation: Wyenandt, A. 2019. Cucurbit downy mildew alert  7/19/19. Plant & Pest Advisory, 19 Jul. https://plant-pest-advisory.rutgers.edu/cucurbit-downy-mildew-alert-7-4-19-2/
  • Type: Other Status: Published Year Published: 2019 Citation: Wyenandt, A. 2019. Cucurbit downy mildew alert  7/4/19. Plant & Pest Advisory, 4 Jul. https://plant-pest-advisory.rutgers.edu/cucurbit-downy-mildew-alert-7-4-19/
  • Type: Other Status: Published Year Published: 2020 Citation: Wyenandt, A. 2020. Controlling basil downy mildew in the greenhouse. Plant & Pest Advisory, 22 Jan. https://plant-pest-advisory.rutgers.edu/controlling-basil-downy-mildew-in-the-greenhouse/
  • Type: Other Status: Published Year Published: 2019 Citation: Wyenandt, C.A., and J.E. Simon. 2019. Four new Rutgers sweet basils available. Morning AgClips 5 Jul. https://www.morningagclips.com/four-new-rutgers-sweet-basil-varieties-available/
  • Type: Other Status: Published Year Published: 2020 Citation: Wyenandt, C.A., Homa, K., and J.E. Simon. 2020. Options for controlling basil downy mildew in field. Morning AgClips, 3 Feb. https://www.morningagclips.com/options-for-controlling-basil-downy-mildew-in-field/


Progress 03/01/18 to 02/28/19

Outputs
Target Audience:Our target audience includes people in the scientific, extension and agricultural communities. Included are other scientists, undergraduate and graduate students and post-docs in plant sciences, plant breeding and plant pathology, crop consultants, extension agents, crop protection industry, growers, shippers, processors and other allied stakeholder industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has been training many people in many field- and laboratory-based agricultural scientific techniques. Field-based techniques include conventional and q-PCR rotorod spore trapping; weather monitoring; establishing and evaluating greenhouse and field fungicide/biopesticide efficacy trials, sentinel plots and cultivar trials including cucurbit, lettuce, spinach and basil production, field plot design, establishment, seed germination, transplanting, maintenance, fungicide application, disease diagnostics/PCR diagnostics, disease evaluation (using the Horfall-Barratt scale), harvesting; canopy cover imaging and analysis; greenhouse plant production; cucumber pollinations and seed processing; cross and self-pollination and drying of basil; organizing breeding populations in the field and prioritizing selections. Laboratory-based techniques include DM pathogen isolations and culture preservation, transfers and cell counting; growth and storage of P. cubensis and P. humuli, DNA extraction, PCR, qPCR, SSR/microsatellite genotyping sequencing analyses and Sanger sequencing; design and development of diagnostic assays including the mitochondrial-based TaqMan Diagnostic assay. Other techniques learned included statistics, network modeling approaches in biological systems, biomathematics on network modeling of cucurbit DM, genomics, bioinformatics, epidemiology, the role of oospores in the pathogen survival and epidemiology; designing, developing and testing of the website for tracking basil DM; coordination and compilation of grower survey results; and preparation of presentations, reports and manuscripts. Post-docs, graduate students and undergraduate students have been trained in the methods and techniques listed above as required for this project under the supervision of the PD and co-PIs and aided by technicians. People who have benefited from this training include post-docs (Alyssa Burkhardt, Shyam Kandel, Alamgir Rahman), graduate students (Julian Bello, Kim D'Arcangelo, Sara Getson, Robert Mattera, Anthony Noto, Maureiq Ojwang, Robert Pyne, Trevor Ruiz, Andy Shirley, Martha Sudermann, Tianxin Wu), undergraduate students (Andew Abdelmalek, Stephanie Aponte, Baylee Carr, Jia-Yi Chan, Brian Change, Abby Dooley, Paul Anthony Edwards, Morgan Gifford, Amber Glenn, Asia Green, Emily Henderson, Clayton Jarvis, Nicholas King, Elizabeth Kogan, Krupali Kothari, Reilly Nakomoto, Han Nguyen, A.J. Noto, Laura Merja, Sara Merrill, Carly Seldow, Trevor Styles, Opal Tam, Jessica Tan, Yazmin Williams), and technicians (Tanner Boss, Allison De Sario, Asia Green, Blair Harlan, Taylere Herrmann, Barry Hughes, Sheila Linderman, Macus Lopez, Patricia Love, Mike Matson, Paige Reeves, Emily Rustad, Sheri Smith, Shane Sullivan, Richard Zimmerman). How have the results been disseminated to communities of interest?Research results have been disseminated via demonstration plots, publications, websites, scientific/extension presentations, webinars, and scientific/extension meetings. NIFA was acknowledged in 16 presentations. Scientific Meetings: Results were disseminated to the scientific community in 11 presentations at scientific meetings, including the Joint International Congress of Plant Pathology/American Phytopathological Society Annual Meeting, Boston, MS, Jul 2018 (3 presentations); Southern Division Annual Meeting, American Phytopathological Society, Fayetteville, AR, 16-18 Feb 2018; 6th International Oomycetes Workshop, Joint International Congress of Plant Pathology/American Phytopathological Society Annual Meeting, Boston, MA, Jul 2018; 5th Flavors, Fragrances and Perception Symposium, Rutgers Douglass Campus Center, New Brunswick, NJ, 30 Nov 2018; Plant Pathology Society of North Carolina Meeting, Raleigh, NC, Oct 2018; Plant Biology Graduate Student Symposium, Rutgers New Jersey Institute for Food, Nutrition and Health, New Brunswick, NJ, 19 Jan 2018; Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, Mar 2018; NCSU Undergraduate Research Symposium, Raleigh, NC, Aug 2018, Institute of Food Technologists Annual Meeting, New Orleans, LA, Jun 2018. Extension Meetings: Research results were disseminated as extension presentations at 26 meetings at the local, regional and national levels, to growers, commodity groups, chemical companies, extension educators, and other agricultural industry stakeholders. Meetings included Pickling Cucumber Session, Great Lakes Fruit, Vegetable and Farm Market Expo, Grand Rapids, MI, 4 Dec 2018; Syngenta Meeting, East Lansing, MI, 29 Mar 2018; Moses Organic Farming Conference, LaCrosse, WI, 22 Feb 2018; Growers' Meeting, Dundee, MI, 8 Feb 2018; Spring County Meeting, Manatee County, FL, 18 Apr 2018; hands-on workshop and field demonstrations, Florida AgExpo, Balm, FL, 2 Nov 2018; SWMREC Winter Extension Meeting, Benton Harbor, MI 23 Jan 2019; California Leafy Greens Research Program (CLGRP) Annual Meeting, Pismo Beach, CA, 27 Mar 2018; California Leafy Greens Research Program Mid-Year Meeting, Salinas, CA, 9 Oct, 2018; Pickle Packers International Conference, Pittsburgh, PA, 19 Apr 2018; Vegetable Breeding Institute Field Days, Freeville, NY, 27 Aug 2018; Empire State Producers Expo, Syracuse, NY, 18 Jan 2018; Grower Meeting, Kreher Family Farms, Batavia, NY, 24 May 2018; Empire Farm Days, Seneca Falls, NY, 8-9 Aug 2018; Cornell Cooperative Extension In-Service Training, Ithaca, NY, 13 Nov 2018; Snyder Farm Open House, Clifford E. & Melda C. Snyder Research and Extension Farm, Pittstown, NJ, 29 Aug 2018; Vegetable Growers Field Day, Rutgers Agricultural Research and Extension Center, Upper Deerfield, NJ, Aug, 2018; Basil Downy Mildew Workshop, New Jersey Vegetable Growers Association and Trade Show, Atlantic City, NJ, 9 Feb 2019; Lettuce Advisory Committee Meeting, Everglades Research and Education Center, Belle Glade, FL, 22 Feb 2018; Lettuce Advisory Committee Meeting, Everglades Research and Education Center, Belle Glade, FL, 3 Oct 2018; New Jersey Vegetable Growers Association Annual Meeting, Atlantic City, NJ, 7 Feb 2019; Lettuce Advisory Committee Meeting, Everglades Research and Education Center, Belle Glade, FL, 27 Feb 2019; BASF Innovations in Agriculture Minority, Raleigh, NC, Apr 2018 (2 presentations); North Carolina Pickle Grower Meeting, Wilson, NC, Mar 2018; Extension Conference, Raleigh, NC, Nov 2018; 33rd Annual Southeast Vegetable and Fruit Expo, Myrtle Beach, NC, Dec 2018. Webinars, open houses, field days, trial tours: Research was featured on 2 events: Downy Mildew Resistant Pickling Cucumber Participatory Plant Breeding Field Evaluation, Plant Pathology Farm, Lansing, MI, 26 Sep 2018; Downy Mildew Resistant Pickling Cucumber Participatory Plant Breeding Field Evaluation, Florida Ag Expo, Wimauma, FL, 7, Nov 2018. What do you plan to do during the next reporting period to accomplish the goals?OBJECTIVE 1: EARLY DETECTION AND RAPID DIAGNOSTICS. co-PI Martin will obtain sporangial dilution series from co-PI Quesada for further testing of diagnostic assay sensitivity and continue to work with PD and co-PIs on technology transfer so they can use the assays in their research. co-PI Quesada and PD Hausbeck will finish processing spore trap samples to compare spore counts with PCR results. Co-PI Gent will provide isolates of P. humuli as requested for comparative testing with assays developed for P. cubensis. co-PI Miles will test a different version of SNP detection for FRAC 11 and FRAC 40 fungicides for rapid-in field isothermal tests of fungicide resistance. co-PIs Miles and Quesada will collaborate on field dissemination of qPCR markers with her graduate student. co-PI Vallad will establish three individual spore traps, an electrostatic, impact rod, and Burkard spore trap, to monitor spore counts and test marker diagnostic assays, and all P. cubensis isolates will be tested with the 10 SSRs and markers for mating type (A1 or A2) in spring and fall. Co-PI Smart will test DNA and frozen sporangia using fungicide resistance diagnostics and provide additional isolates as needed. Co-PI Vallad will collect P. cubensis isolates from FL and test for resistance to fluopicolide, cyazofamid, and oxathiapripolin using small plot trials, limited in vivo testing, and available molecular markers. OBJECTIVE 2: TRACKING PATHOGEN DISPERSAL AND SURVIVAL. Co-PI Ojiambo will refine and update the national basil DM tracking website based on stakeholder needs and create a link to assess DM reports within the cucurbit DM ipmPIPE site. PD Hausbeck and co-PIs Quesada, Smart, Smith, Vallad, and Wyenandt will continue sentinel plot deployment to track movement of DM disease on cucurbits and basil. PD Hausbeck and co-PI Vallad will continue to monitor airborne sporangia and compare efficacy of molecular diagnostics via PCR rotorod spore traps versus conventional traps. Co-PI Vallad will also compare an electrostatic spore trap. Co-PIs Gent and Ojiambo will investigate alternative modeling frameworks that can accommodate sparse data typical of the cucurbit DM PIPE system, with the intent to predict the epidemic front of spread for DM from historical data sets and determine whether one or multiple inoculum sources are likely responsible for disease outbreaks in northern latitudes. PD Hausbeck and co-PIs Smart, Vallad and Wyenandt will continue to monitor duration of DM control with fungicide efficacy trials. OBJECTIVE 3: DEVELOPING DM-RESISTANT CROPS. co-PI Mazourek will increase seed from 2018 field selections of the DM-resistant cucumber breeding project. Trials will be conducted in 2019 with all promising selection from 2018 evaluated in a replicated complete block design. co-PI Mou will plant seed harvested from 23 DM-resistant spinach populations, resistant/susceptible controls, and evaluate DM incidence. Resistant plants will be selected and transplanted into isolators for seed production. This cycle of recurrent selection will continue until a satisfactory level of resistance is achieved. Populations will be field tested for DM resistance, uniformity, horticultural traits, and yield prior to public release. co-PI Simon will evaluate new basil seed for DM and newly available DM-resistant basil cultivars for field resistance to DM. New crosses and selections of basil will continue to be evaluated for field performance and DM resistance. Basil DNA will continue to be extracted and all lines will be sequenced and genotyped with SNPs and SSR markers. co-PI Mazourek will increase the 2018 selections of the participatory cucumber plant breeding project and PD Hausbeck and co-PI Agehara will establish them in 2019 field trials in MI and FL to evaluate/select the breeding lines with commercial growers and extension agents. OBJECTIVE 4: COST/BENEFIT RATIOS AND GROWER ADOPTION. PD and co-PIs will continue to administer surveys prepared by co-PI Govindasamy and collect data as needed. OBJECTIVE 5: OUTREACH AND ADOPTION. co-PI Ojiambo will continue to coordinate the monitoring and reporting of disease outbreaks using sentinel plots and uploaded disease forecasts and disease alerts to DM stakeholders during the growing season. PD Hausbeck and co-PIs Agehara, Quesada, Raid, Smart, Smith and Vallad will continue to report DM outbreaks to the ipmPIPE monitoring/forecasting system and train growers and extension personnel in the use of this system. Grower outreach educational programs will be planned for the 2019 growing season. PD Hausbeck and co-PIs Quesada, Raid, Smart, Vallad and Wyenandt will test breeding lines/varieties in their regions for DM resistance as needed. PD Hausbeck and co-PIs Agehara and Smith will evaluate breeding lines/varieties in their region for horticultural traits. PD Hausbeck and co-PIs will conduct basil seed testing experiments, including testing commercial seed for the presence of DM develop/disseminate educational materials on the importance of clean seed. PD Hausbeck and co-PIs Quesada, Raid, Smart and Wyenandt will continue developing and disseminating field spray recommendations based on regional trials. PD Hausbeck and co-PIs Agehara, Goldy, Quesada, Raid, Smart, Smith, Vallad and Wyenandt will continue developing social media and posting frequent updates to advise growers. OBJECTIVE 6: ATTRACT STUDENTS TO AGRICULTURAL SCIENCES. PD and all PIs will continue to recruit post-docs, graduate and undergraduate students as needed, and will continue to mentor them in scientific techniques needed to participate in project research. The summer research program for undergrads will continue to be implemented; we expect at least 4 students to participate in a Research Experiences for Undergraduates program, one at NCSU with co-PI Quesada and three at MSU with co-PI Miles.

Impacts
What was accomplished under these goals? OBJECTIVE 1: EARLY DETECTION AND RAPID DIAGNOSTICS. Co-PI Martin further tested the multiplexed mitochondrial markers for detection of P. cubensis and P. humuli and shared the assays with co-PIs Quesada, Vallad, Smart. Co-PI Miles conducted genomic analysis of P. cubensis and identified putative loci resistance to FRAC 49. PCR detection systems for FRAC 11 and FRAC 40 resistance have been refined. Co-PI Miles showed that loop-mediated isothermal amplification (LAMP) shows more promise than recombinant polymerase amplification (RPA) for in-field single-nucleotide polymorphism (SNP) detection. Co-PI Miles identified the resistance mechanism to carboxylic acid amide (CAA) and quinone outside inhibitor (QoI) fungicides as two SNPs at the cellulose synthase 3 gene (CesA3 at amino acid positions 1105 and 1109) and in the cytochrome oxidase b gene (Cyt b at amino acid position 143), respectively, in P. cubensis isolates. Co-PI Miles developed a molecular assay to identify the 1105 and 143 SNPs, and determine the occurrence of CAA and Cyt B resistance. Results from the CesA3 and Cyt b sequence alignments showed that resistance was most prevalent in cucumber P. cubensis isolates. OBJECTIVE 2: TRACKING PATHOGEN DISPERSAL AND SURVIVAL. Co-PI Ojiambo discussed developing a framework to incorporate the basil DM monitoring site with the cucurbit DM ipmPIPE infrastructure with Cornell University. co-PI Ojiambo completed analyses to relate size of initial disease focus on final disease extent. The consistency of the power law model in determining the rate of epidemic development was determined. The cucurbit DM ipmPIPE continued to provide information on the disease spread during the season. Seven cucurbits and basil were planted in sentinel plots (53 total) for monitoring DM during the reporting year, including plots in MI, NC, NY, FL and NJ by PD Hausbeck and co-PIs Quesada, Smart, Vallad, Wyenandt. Roto-rod spore traps provided by co-PI Quesada were deployed in NC, MI and FL by co-PIs Quesada, Vallad and PD Hausbeck. PD Hausbeck deployed conventional Burkard for comparison with roto-rod spore traps in MI. DNA of P. cubensis and P. humuli DNA were detected by Burkard spore traps, while DNA of P. cubensis only was detected by rotorod spore traps. qPCR results were corroborated by reanalyzing a random set of samples with multiplexed mitochondrial marker system developed for P. cubensis and P. humuli by co-PI Martin, which differentiated the two clades of P. cubensis. PD Hausbeck and co-PIs Ojiambo, Smart, Vallad established fungicide efficacy trials; cucurbit DM trials in MI, NC, NY and FL and a basil DM trial in NC. Treatments evaluated included conventional and organic fungicides, products applied alone or in combination/alternation, and timing of initiation of the spray program. OBJECTIVE 3: DEVELOPING DM-RESISTANT CROPS. Co-PI Mazourek evaluated >3,000 cucumber plants from 47 pedigree F4 families and recurrent selection populations under natural DM pressure in 2018. Plants (28) were selected for strong DM resistance and good horticultural and fruit/consumer characteristics (i.e. earliness and fruit shape) to be advanced over the winter greenhouse season. Ten 2017 selections were evaluated in a 2018 yield trial; all were improved compared to susceptible Vlaspik. Co-PI Mou crossed open-pollinated DM-resistant spinach cultivars with different DM-resistance genes to combine their resistances and planted progenies in the field; resistant plants were selected for open pollination and seed production. The USDA spinach germplasm collection (386 accessions) were planted in the field to map and find molecular markers for the field resistance to DM. Co-PI Simon continued the basil DM breeding program. Seed from the F7 generation was planted in Apr 2018 and the resulting F8 plants from the 113 recombinant inbred lines (RILs) and 2 parents were transplanted in 2 field locations in NJ and grown into adulthood to produce seed for the next generation. PD Hausbeck and co-PI Agehara established participatory plant breeding trials of DM-resistant cucumbers in MI and FL, including 250 seeds of the intermated pickle breeding populations from their respective sites the previous year, and 50 seeds each of 4 individual lines selected from their sites the previous year. Plants with preferred characteristics were selected by co-PI Mazourek and pathologists, growers, processors, and extension specialists. Disease-free cuttings were taken to be intermated for the recurrent breeding population, and also within those selected from the individual breeding lines; 25 selections were rooted from MI and 11 from FL. Co-PI Quesada tested 8 cultivars and Plant Introduction cucumber varieties for DM resistance in NC. PI 330628 and PI 197088 had significantly less DM than other cultivars, while Peacemaker had the highest yield. Co-PI Smith established a cucumber variety trial of the DM-resistant NY varieties; however, fruit were commercially unacceptable as they were too short. OBJECTIVE 4: COST/BENEFIT RATIOS AND GROWER ADOPTION. Co-PI Govindasamy developed a producer survey to collect DM data. It was administered to vegetable growers with the help of the PD and co-PIs. OBJECTIVE 5: OUTREACH AND ADOPTION. As part of the cucurbit DM monitoring system for the 2018 season, sentinel plots were established in the eastern U.S. and scouted weekly until disease was observed. DM disease outbreaks were reported to the cucurbit ipmPIPE website (http://cdm.ipmpipe.org/) in 150 counties in 2018. Disease forecasting was conducted 2-3 times a week during the growing season from March to October, 2018. This information was available at the cucurbit ipmPIPE website. Regional reports of cucurbit DM occurrences to the cucurbit ipmPIPE website was kept up to date by the PD and co-PIs, ensuring accurate information was available to growers during 2018. Open houses, field days and trial tours also occurred in 2018 to alert and inform growers, industry and extension personnel of results of 2018 research (summarized in "How have the results been disseminated to communities of interest"). Results of 2018 trials testing cultivars for DM resistance allows evaluation of the performance of these cultivars over geographical areas and seasonal weather differences, generating valuable information for growers. The new Rutgers DM-resistant sweet basil cultivars, Passion, Devotion, Obsession and Thunderstruck, were made available to commercial growers. Fungicide efficacy trials provided growers recommendations for managing DM that are up to date and tailored to their regions. Results were communicated via social media during the 2018 growing season as they occurred to ensure that growers received the latest DM information and recommendations in a timely manner. OBJECTIVE 6: ATTRACT STUDENTS TO AGRICULTURAL SCIENCES. Katelyn Goldenhar completed her M.S. (Thesis: Management of cucumber DM with fungicides and host resistance) under the direction of PD Hausbeck. The PD and co-PIs have recruited 3 post-docs, 8 graduate students, and 25 undergraduate students to work on this project in the past year. Two students attended the Research Experiences for Undergraduates program at CSUMB that were partially or fully funded by this grant. Ross Hatlen was recently accepted into a Plant Pathology PhD program at Michigan State University starting in Fall 2019. This proposal has trained a number of undergraduate students and attracted them to plant sciences in general. Twelve students have gone on to graduate school in plant sciences. These twelve students either received the modified plant science lecture content in three different courses at CSUMB or attended the REU program at Michigan State University, North Carolina State University or Cornell. Several students also are currently working professionally in the agricultural industry in the Monterey Bay area at large agricultural companies (e.g. Sakata Seed, Driscoll's, Tanamura and Antle).

Publications

  • Type: Theses/Dissertations Status: Published Year Published: 2018 Citation: Goldenhar, K.E. 2018. Management of cucumber downy mildew with fungicides and host resistance. M.S. Thesis, Michigan State University, East Lansing.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Fletcher, K., Klosterman, S.J., Derevnina, L., Martin, F., Bertier, L.D., Koike, S., Reyes-Chin-Wo, S., Mou, B., and Michelmore, R. 2018. Comparative genomics of downy mildews reveals potential adaptations to biotrophy. BMC Genomics 19:851.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Qin, J., Shi, A., Mou, B., Grusak, M.A., Weng, Y., Ravelombola, W., Bhattarai, G., Dong, L., and Yang, W. 2017. Genetic diversity and association mapping of mineral element concentrations in spinach leaves. BMC Genomics 18:941
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Shi, A., Qin, J., Mou, B., Correll, J., Weng, Y., Brenner, D., Feng, C., Motes, D., Yang, W., Dong, L., Bhattarai, G., and Ravelombola, W. 2017. Genetic diversity and population structure analysis of spinach by single-nucleotide polymorphisms identified through genotyping-by-sequencing. PLoS ONE 12(11):e0188745.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wyenandt, C.A., McGrath, M.T., Everts, K.E., Rideout, S.L., Gugino, B.K., and Kleczewski, N.M. 2018. Fungicide resistance management guidelines for cucurbit downy and powdery mildew control in the mid-Atlantic and Northeast regions of the United States in 2018. Plant Health Progress 19:34-36.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Crandall, S.G., Rahman, A., Quesada-Ocampo, L.M., Martin, F.N., Bilodeau, G. J., and Miles, T.D. 2018. Advances in diagnostics of downy mildews: lessons learned from other oomycetes and future challenges. Plant Disease 102:265-275.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2018. Evaluation of fungicides for control of downy mildew on cucumber II, Clinton 2017. Plant Disease Management Reports 12:V096.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2018. Evaluation of fungicides for control of downy mildew on cucumber, Clinton 2017. Plant Disease Management Reports 12:V097.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2018. Evaluation of cultivars in combination with fungicides for control of downy mildew and yield effects on cucumber, Clinton 2017. Plant Disease Management Reports 12:V098.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Pyne, R.M., Honig, J.A., Vaiciunas, J., Wyenandt, C.A., and Simon, J.E. 2018. Population structure, genetic diversity and downy mildew resistance among Ocimum species germplasm. BMC Plant Biology 18(1):p.69.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wyenandt, C.A., Maimone, L.R., Homa, K., Madeiras, A.M., Wick, R., and Simon, J.E. 2018. Detection of the downy mildew pathogen on seed of basil following field infection in southern New Jersey. HortTechnology 28(5):637-641.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wyenandt, C.A., McGrath, M.T., Everts, K.E., Rideout, S.L., Gugino, B.K., and Kleczewski, N. 2018. Fungicide resistance management guidelines for cucurbit downy and powdery mildew in the mid-Atlantic and northeast regions of the United States in 2018. Plant Heath Progress 19:34-36.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Wyenandt, C.A., Maimone L.R., Homa, K., Madeiras, A.M., Wick, R.L., and Simon, J.E. 2018. Detection of downy mildew (Peronospora belbahrii) in seed of basil (Ocimum spp.) following field epidemics in southern New Jersey. HortTechnology 28(5):637-641.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, M.L., DArcangelo, K., Collins, H., and Quesada-Ocampo, L.M. 2018. Evaluation of fungicides and cultivars for control of downy mildew on cucumber, Kinston 2017. Plant Disease Management Reports 12:V116.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: DArcangelo, K.N., Adams, M.L., and Quesada-Ocampo, L.M. 2018. Evaluation of cultivars for control of downy mildew on cucumber, Kinston 2017. Plant Disease Management Reports 12:V126.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, M.L., Collins, H., and Quesada-Ocampo, L.M. 2018. Evaluation of fungicides for control of downy mildew of winter squash, Kinston 2017. Plant Disease Management Reports 12:V148.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: DArcangelo, K.N., Adams, M.L., and Quesada-Ocampo, L.M. 2018. Evaluation of cultivars for control of downy mildew on squash, Kinston 2017. Plant Disease Management Reports 12:V151.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Hausbeck, M.K., and Goldenhar, K. 2017. Cucumber disease control in 2017. Pages 6-11 in: Pickling Cucumber Session Summaries, Great Lakes Fruit, Vegetable and Farm Market Expo, Grand Rapids, MI, Dec. Online at https://glexpo.com/wp-content/uploads/2018/12/Pickling-Cucumber.pdf.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Smart, C.D., and Lange, H. 2018. Update on cucurbit diseases. Proceedings of the Empire State Producers Expo, Syracuse, NY. Jan 2018. Online.
  • Type: Other Status: Published Year Published: 2018 Citation: Hausbeck, M.K., and Linderman, S.D. 2018. Monitoring and managing cucurbit downy mildew. Online at https://veggies.msu.edu/wp-content/uploads/2017/05/FS_Cucurbit_DM_2017.pdf.
  • Type: Other Status: Published Year Published: 2018 Citation: Hausbeck, M. 2018. Downy mildew is found in Michigan cucumbers. Michigan State University Extension News for AgricultureVegetables: 25 Jul. Online at https://www.canr.msu.edu/news/downy-mildew-found-in-michigan-cucumbers-in-2018.
  • Type: Other Status: Published Year Published: 2017 Citation: DArcangelo, K., Miles, T., and Quesada-Ocampo, L.M. 2017. Occurrence of fungicide resistance in Pseudoperonospora cubensis populations causing cucurbit downy mildew in commercial and wild hosts. Abstract. Phytopathology 107(12S):S5.63.
  • Type: Other Status: Published Year Published: 2017 Citation: Mandal, M.K., Ikerd, J., Wallace, E., Grace, R., Turechek, W., Quesada-Ocampo, L.M., and Kousik, C. 2017. Population biology of the downy mildew pathogen on tolerant and susceptible cucumber in the southeastern United States. Abstract. Phytopathology 107(12S):S5.127.
  • Type: Other Status: Published Year Published: 2018 Citation: DArcangelo, K., Miles, T., and Quesada-Ocampo, L.M. 2018. Pseudoperonospora cubensis populations infecting wild and commercial cucurbit hosts display host-specific sensitivity to fungicides. Abstract. Phytopathology 108:S1.1.
  • Type: Other Status: Published Year Published: 2018 Citation: Hausbeck, M. 2018. Time for downy mildew protectant sprays for cucumber. Michigan State University Extension News for AgricultureVegetables: 26 Jun. Online at https://www.canr.msu.edu/news/time-for-downy-mildew-protectant-sprays-for-cucumbers.
  • Type: Other Status: Published Year Published: 2018 Citation: Rahman, A., and Quesada-Ocampo, L.M. 2018. Biosurveillance for precision disease management of Pseudoperonospora cubensis, the cucurbit downy mildew pathogen. Abstract. Phytopathology 108:S1.52.
  • Type: Other Status: Published Year Published: 2018 Citation: Shirley, A.M., and Vallad, G. 2018. Establishing residual activity of current fungicides against downy mildew on cucumber. Abstract. Phytopathology 108(12S):S2.9.
  • Type: Other Status: Published Year Published: 2018 Citation: Quesada-Ocampo, L.M. 2018. Basil downy mildew found in North Carolina. Extension Plant Pathology Portal, June 26, 2018. Online at https://plantpathology.ces.ncsu.edu/2018/06/basil-downy-mildew-found-in-north-carolina/.
  • Type: Other Status: Published Year Published: 2018 Citation: Quesada-Ocampo, L.M., and DArcangelo, K. 2018. Cucumber downy mildew reported in North Carolina. Extension Plant Pathology Portal, June 11, 2018. Online at https://plantpathology.ces.ncsu.edu/2018/05/cucumber-downy-mildew-reported-in-north-carolina-3/.


Progress 03/01/17 to 02/28/18

Outputs
Target Audience:Our target audience includes people in the scientific, extension and agricultural communities. Included are other scientists, undergraduate and graduate students and post-docs in plant sciences, plant breeding and plant pathology, crop consultants, extension agents, crop protection industry, growers, shippers, processors and other allied stakeholder industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has been training many people in many field- and laboratory-based agricultural scientific techniques. Field-based techniques include spore trapping; weather monitoring; field plot design, establishment, seed germination, transplanting, maintenance, fungicide application, disease evaluation (using the Horfall-Barratt scale), harvesting; establishing efficacy trials and sentinel plots; canopy cover imaging and analysis; greenhouse plant production; cucumber pollinations and seed processing; cross and self-pollination and drying of basil; organizing breeding populations in the field and prioritizing selections. Laboratory-based techniques include DM pathogen isolations, transfers and cell counting; DNA extraction, PCR, qPCR, SSR/microsatellite genotyping sequencing analyses and Sanger sequencing; development of diagnostic tools and the mitochondrial-based TaqMan Diagnostic assay. Other techniques learned included statistics, biomathematics, genomics, bioinformatics, epidemiology, preparation of presentations, reports and manuscripts; designing, developing and testing of the website for tracking basil DM; coordination and compilation of grower surveys. People who have benefited from this training include post-docs (Surendran Arumugam, Shyam Kandel, Alamgir Rahman), graduate students (Lauren Brzozowsi, Kim D'Arcangelo, Katelyn Goldenhar, Kathryn Homa, Robert Mattera, Maureiq Ojwang, Robert Pyne, Trevor Ruiz, Andy Shirley, Greg Vogel, Jesse Yamagata), undergraduate students (Andew Abdelmalek, Madison Ahmad, Stephanie Aponte, Sierra Barnes, Tanner Boss, Ali Cala, Baylee Carr, Jordan Castellari, Jia-yi Chan, Brian Change, Howard Chhen, Samuel Cude, Gina Dabbah, Phi Diep, Abby Dooley, Paul Anthony Edwards, Philip Engelgeau, Morgan Gifford, Amber Glenn, Asia Green, Jolie Habashy, Thilani Jayakody, Elizabeth Kogan, Courtney Lepping, Devin Linderman, Adrian Maldonado, Anastasia Matta, Laura Merja, Sarah Merrill, A.J. Noto, Lisa Panichelli, Gabriel Ramirez, Carly Seldow, Aidan Shands, Opal Tam, Miranda Weatherly, Darius Wells), and technicians (Mariami Bekauri, Alyssa Burkhardt, Chris DelCastillo, Scott Hughes, Steve Kalb, Emi Kuroiwa, Holly Lange, Sheila Linderman, Evan McClenthan, Marina Ramon, Paige Reeves). How have the results been disseminated to communities of interest?Research results have been disseminated via demonstration plots, publications, websites, scientific/extension presentations, webinars, and scientific/extension meetings. NIFA was acknowledged in 16 presentations. Scientific Meetings: Results were disseminated to the scientific community in 14 presentations at scientific meetings, including the American Phytopathological Society Meeting (4 presentations), San Antonio, TX, Aug; Oomycete Molecular Genetics Network Annual Meeting (2 presentations), Asilomar, CA, 11-14 Mar; 4th Annual Flavors, Fragrances & Perception Symposium (2 presentations), Rutgers University, New Brunswick, NJ, 8 Jun; American Phytopathological Society Potomac Division Meeting, Morgantown, WV, Mar; American Phytopathological Society Southern Division Meeting, College Station, TX, 18-20 Feb; 8th Annual Conference, American Council for Medicinally Active Plants (ACMAP), Clemson University, SC, 22 Jun; Rutgers 240 Breeding Celebration and Luncheon, New Brunswick, NJ, 4 Nov; Scientific Seminar, Universidad de los Andes, Bogota, Colombia; and NCSU Undergraduate Research Symposium. Raleigh, NC, Aug. Extension Meetings: Research results were disseminated as extension presentations at 29 meetings at the local, regional and national levels, to growers, commodity groups, chemical companies, extension educators, and other agricultural industry stakeholders. Meetings included the Michigan Agribusiness Association Meeting, Lansing, 11 Jan; Agricultural Extension Consultants Breakfast, Hart, MI; Michigan State University Extension and AgBioResearch State CouncilMeeting, Lansing, MI, Mar; Organic Growers' Meeting, East Lansing, MI, 9 Mar; Wisconsin Fresh Fruit and Vegetable Conference, Wisconsin Dells, WI, 23 Jan; Seminis Meeting, Charleston, SC, Oct; Southwest Hort Days, Benton Harbor, MI, Feb; Bay Area Growers Extension Meeting, Bay City, MI, Jan; Western NY Field Days, Portland, NY, 31 Aug; Empire Farm Days, Seneca Falls, NY, 8 Aug; Agricultural Career Days, Finger Lakes, NY, 26 Apr; California Leafy Greens Research Program (CLGRP) Annual Meeting, Pismo Beach, CA, 14 Mar; California Leafy Greens Research Program Mid-Year Meeting, Salinas, CA, 10 Oct; Florida Ag Expo, Wimauma, FL, 8 Nov; North Country Willsboro Research Farm Summer Growers Meeting, Willsboro, NY, 4 Aug; Canton Summer Growers Meeting, Canton, NY, 3 Aug, Western NY Veg Growers Meeting, Lockport, NY, 1 Mar; Discovering Geneva Community Event, Geneva, NY, 27 Feb; Managing cucurbit downy mildew. Empire State Producers Expo, Syracuse, NY, 20 Jan; NY Fun on the Farm, 30 Sep; 2016 Crop Consultant Meeting, Syracuse, NY, 1 Dec; Florida AgExpo, Wimauma, FL, 8 Nov; North Jersey Vegetable Growers Meeting, Flemington, NJ, 23 Feb; NJVGA Annual Vegetable Growers Meeting, Atlantic City, NJ, 8 Feb; NJ-NOFA Annual Meeting, New Brunswick, NJ, 28 Jan; New Holland Vegetable Growers Meeting, New Holland, PA, 16 Jan; NJVGA Annual Vegetable Growers Meeting (3 presentations), Atlantic City, NJ, 8 Feb. Webinars, open houses, field days, trial tours: Research was featured on 3 webinars and 19 open houses/field days/trial tours: Cucumber Breeding Trial, Field Day, Florida Ag Expo, Wimauma, FL, 8 Nov; Participatory Cucumber Downy Mildew Trial, Michigan State University, Lansing, MI, 22 Sep; Downy Mildew Pickling Cucumber Field Day, Ransomville, NY, 26 Sep; Cornell Vegetable Breeding Institute Field Days, Freeville, NY, 28 Aug; Grower Field Day, Everglades Research and Education Center, Belle Glade, FL, 24 Mar; Open House, Everglades Research and Education Center, Belle Glade, FL, 6 Apr; Grower Field Day, Everglades Research and Education Center, Belle Glade, FL, 21 Apr; Dept. of Agriculture tour, Everglades Research and Education Center, Belle Glade, FL, 16 May; Plant pathology issues, Everglades Research and Education Center, Belle Glade, FL; Lettuce Advisory Meeting, Everglades Research and Education Center, Belle Glade, FL, 4 Oct; First Detector Meeting, Everglades Research and Education Center, Belle Glade, FL, 1 Nov; Basil DM variety breeding evaluations, Rutgers University, New Brunswick, NJ, 11-12 Aug; Open House Everglades REC, Everglades Research and Education Center, Belle Glade, FL, 6 Apr; Glades Day School Field Trip, Everglades Research and Education Center, Belle Glade, FL, 20 Apr; Pickling Cucumber Breeding Demonstration, Niagara County, NY; Cucumber Downy Mildew Field Demonstration, Florida Ag Expo, Wimauma, FL, 8 Nov; Cucumber Downy Mildew Fungicides Field Demonstration, Florida Ag Expo, Wimauma, FL, 8 Nov. New Lectures in Plant Breeding (Plant Biology, graduate course) and in Sensory Sciences (Dept. Food Science, undergraduate course) developed and presented in 2017 from this project initiative and with NIFA acknowledged: Breeding for disease resistance, nutrition and aroma, Case study with sweet basil and downy mildew, Plant Breeding, Spring Semester, 2017; Chemistry of aromas, Case study with sweet basil, Sensory Sciences, Spring Semester, 2017. What do you plan to do during the next reporting period to accomplish the goals?OBJECTIVE 1. Early detection and rapid diagnostics. co-PI Martin will evaluate the sensitivity and specificity of the diagnostic assay developed in his lab compared to the nuclear assay from co-PI Quesada's lab and the cox2 marker from co-PI Smart's lab using a standardized set of DNAs (same DNAs will be used in all three labs).co-PI Martin has provided the internal controls to co-PI Vallad and will assist his program in using molecular assays for pathogen detection.co-PI Quesada will continue developing a P. cubensis detection system via spore traps using designed nuclear and mitochondrial markers in collaboration with co-PI Martin. co-PI Miles will continue to develop assays to detect resistance to fungicides, including active ingredients with differing FRAC codes. co-PIs Miles and Quesada will collaborate on developing the TaqMan marker system to detect fungicide resistance. We currently have 2 markers that will be used to develop RPA assays for FRAC 11 and 40 fungicides. Ongoing research is to identify the loci associated with other fungicide chemistries.PD Hausbeck and co-PIs will provide isolates and test markers for all diagnostic assays developed in Objective 1. Rigorous field testing of the molecular assays in comparison to traditional spore trapping will take place in grower cooperator fields. OBJECTIVE 2. Tracking pathogen dispersal and survival. co-PI Ojiambo will continue ipmPIPE administration, in particular coordinating the monitoring and reporting of disease outbreaks using sentinel plots and providing maintenance for the basil DM website. PD Hausbeck and co-PIs Quesada, Smart, Smith, Vallad and Wyenandt will continue sentinel plot deployment track movement of DM disease on cucurbits and basil.PD Hausbeck and co-PIs Quesada and Vallad will deploy spore traps and test seeds. co-PI Ojiambo will continue synthesizing reports of disease outbreaks to generate disease alerts for the cucurbit stakeholders. co-PI Ojiambo will work with co-PIs Bhattacharyya and Gent to develop a framework and develop a model for network transmission of cucurbit DM. co-PIs Bhattacharyya and Gent will examine historical epidemic and weather data to form networks of DM spread, distinguish between DM due to disease spread or new occurrences, and provide a sampling scheme of identifying infectious source fields by finding locations critical for disease spread. PD Hausbeck and co-PIs Quesada, Smart, Vallad and Wyenandt will continue to collect data for modeling grower alerts.PD Hausbeck and co-PIs Quesada, Ojiambo and Vallad will continue research of DM survival between growing seasons. Proposed research will include planting wild cucurbits in sentinel plots to determine the potential role in the overwintering of the cucumber DM pathogen. co-PI Ojiambo will start analyzing relevant collected from field trials to quantify DM control in fungicide treated and untreated plots and analyze data using generalized linear mixed model to derive corresponding control functions. PD Hausbeck and co-PIs Smart, Vallad and Wyenandt will continue to establish yearly field experiments to obtain data on the efficacy of DM fungicides including new, unlabeled products. OBJECTIVE 3. Developing DM-resistant crops. co-PI Mazourek will continue cucumber breeding by selfing 36 individuals from pedigree and recurrent selection populations for 2018 replicated progeny family plots, and 15 were chosen from three populations to continue intermating for population improvement. Top performing lines for yield, fruit quality and resistance will also be backcrossed to preferred processing types, fresh eating types and seedless types for hybrid evaluation and populations development.co-PI Mou will plant seeds harvested from 26 spinach populations between 80-inch beds of susceptible 'Viroflay' which will provide natural inoculants of DM pathogen. Leaf samples with sporulation of the pathogen will be collected to identify pathogen races using differential hosts. DM-resistant plants will be selected and transplanted into isolators for seed production. This cycle of recurrent selection will continue until a satisfactory level of resistance is achieved.co-PI Simon will evaluate Rutgers DM resistant sweet basil lines for resistance/tolerance in field trials and will continue to construct large backcross populations to overcome the frequency of recessive resistance-reducing alleles and achieve reasonable numbers of resistant individuals. Pedigree breeding will be continued to the F9 generations to fix dominant, DM resistant loci, eliminate recessive alleles conferring susceptibility, and simultaneously select for desirable horticultural traits. Studies on the genetic polymorphism will continue to allow for mapping of QTLs to identify genes involved in DM resistance.Participatory cucumber breeding: co-PI Mazourek will intermate and self regionally selection cuttings between locations. Depending on seed availability, improved populations will be redistributed to PD Hausbeck and co-PIs Agehara, Goldy, Quesada, Smart and Smith again in 2018 for another round of selection along with progeny rows from regional grower selections. OBJECTIVE 4. Cost/benefit ratios and grower adoption. co-PI Govindasamy will continue to work on baseline survey data to tabulate and analyze results. Enterprise budgets with five sections will be developed for the selected crops. co-PI Govindasamy will document behavioral, social and environmental outcomes by evaluating grower preference and the impact of DM on selected crops. Profitability of the farms growing these selected crops will be estimated based on both primary and secondary data collection process. PD Hausbeck and co-PIs Quesada, Raid, Smart, Smith, Vallad and Wyenandt will continue to provide input for this objective. OBJECTIVE 5. Outreach and adoption. co-PI Ojiambo will continue to coordinate the monitoring and reporting of disease outbreaks using sentinel plots and availing disease forecasts and disease alerts to DM stakeholders during the growing season. PD Hausbeck and co-PIs Agehara, Quesada, Raid, Smart, Smith and Vallad will continue to report DM outbreaks to the ipmPIPE monitoring/forecasting system and train growers and extension personnel in the use of this system. Grower outreach educational programs will be planned for the 2018 growing season. PD Hausbeck and co-PIs Quesada, Raid, Smart, Vallad and Wyenandt will test breeding lines/varieties in their region for DM resistance. co-PIs Agehara, Goldy and Smith will evaluate breeding lines/varieties in their region for horticultural traits.PD Hausbeck and co-PIs Quesada, Vallad and Wyenandt will conduct basil seed testing experiments, including testing commercial seed for the presence of DM using RT-PCR to determine how much infested seed maybe entering commercial use. PD Hausbeck and co-PIs Agehara, Goldy, Quesada, Raid, Smart, Vallad and Wyenandt will develop/disseminate educational materials on the importance of clean seed.PD Hausbeck and co-PIs Quesada, Raid, Smart and Wyenandt will continue developing and disseminating field spray recommendations based on regional trials. PD Hausbeck and co-PIs Agehara, Goldy, Quesada, Raid, Smart, Smith, Vallad and Wyenandt will continue developing social media and posting frequent updates. OBJECTIVE 6. Attract students to ag sciences. PD and all PIs will continue to mentor post-docs, graduate and undergraduate students who participate in project research. The summer research program for undergrads will continue to be implemented.

Impacts
What was accomplished under these goals? Objective 1. Early detection and rapid diagnostics. co-PI Martin's mitochondrial haplotype assay has been finalized is ready to be used to support P. cubensis population studies. The LNA probe-based qPCR protocol developed by co-PIs Gent, Smart for differentiating P. cubensis and P. humuli has been completed. LNA probes were specific and sensitive, detecting as few as 10 sporangia for both species and as little as 1 fg P. cubensis total DNA and 10 fg P. humuli total DNA. The HRM assay correctly diagnosed all tested isolates as well as symptomatic plants collected in the field. It was successfully tested in MI. The LNA probe-based qPCR assay tested in MI and was able to detect DNA from extractions made out of solutions containing 20, 50, 100, 200 and 300 sporangia. The relationship between sporangial counts and Cq using the DNA extracted from the five different sporangial counts spiked in the matrix was significant and the assay exhibited a linear response with a R2 value of 0.629, suggesting that changes in sporangial counts are related to changes in Cq values. Roto-rod traps for detecting cucurbit DM spores were tested in NC, MI and FL for the 2017 growing season. Analysis of the season's worth of sampling rods will provide data to refine development of the traps. co-PI Quesada found the modified Phenol-chloroform method and MO-BIO kit performed similarly, both of which were better than QIAmp DNA mini kit in extracting DNA. Two different organic solvents, chloroform and ethyl acetate, are able to disperse silicon grease and neither solvent are known to have any detrimental effect on DNA molecules. co-PIs Quesada and Miles have developed a TaqMan assay for fungicide resistance diagnostics to test carboxylic acid amide FRAC code 40) and quinone outside inhibitor (FRAC code 11) fungicides. Further work will include fungicides of other FRAC codes. Objective 2. Tracking pathogen dispersal and survival. co-PI Ojiambo has created a functional basil DM forecasting website which will become publicly available in 2018.co-PI Ojiambo established a framework for incorporating important weather variables into a network model for disease transmission. Confirmed DM-infected cucurbit crops from the 2017 sentinel plots established by PD Hausbeck and co-PIs Quesada, Ojiambo, Smart, Smith, Vallad were observed and reported as part of the cucurbit DM monitoring network. Cucurbit DM occurrences and forecasting the movement of the disease were available publicly to growers at the cucurbit DM ipmPIPE website during the 2017 growing season. Based on this year's fungicide efficacy trials established by PD Hausbeck and co-PIs, Orondis products are highly effective and have been added to their fungicide recommendations to growers for use in a DM program along with other products shown to be effective in previous trials. Objective 3. Developing DM-resistant crops. Another season of breeding using different techniques by co-PIs Mazourek, Mou and Simon in 2017 has produced cucumber, spinach and basil plant populations with increased DM resistance and good horticultural traits to be used for next season's breeding effort to further refine the crops. Under co-PI Simon's supervision, work was completed on developing a first linkage map for DM resistance for sweet basil. The map was constructed using 42 EST-SSR and 1,847 SNP markers spanning 3,030.9 cM. Results obtained from nested Bayesian model-based clustering, analysis of molecular variance (AMOVA), and unweighted pair group method using arithmetic average (UPGMA) analyses were synergized to provide an updated phylogeny of the Ocimum genus. One year of participatory plant breeding of DM-resistant cucumbers was accomplished in MI and NY and is in progress in FL. Desirable plants were selected with the input of growers, growers, processors, extension specialists and pathologists. co-PI Mazourek took disease-free cuttings in MI and NY for producing the next generation of plants for 2018. Objective 4. Cost/benefit ratios, grower adoption. Basil survey results compiled by co-PI Govindasamy indicate that, on average, growers lost 52% of basil yield due to DM, a loss of $15,600 or more per season in greenhouse farming with 2304 square feet. Growers indicated that 40% of them are using more fungicides on basil now when compared to about 5 years ago. Actinovate AG, Serenade MAX, and Oxidate were the most popular organic fungicides, and Quadris and Ranman the most popular conventional fungicides in use. Growers found agricultural extension services to be very helpful, as program technicians often offer the necessary tools needed for the producers' success. Objective 5. Outreach and adoption. As part of the cucurbit DM monitoring system for the 2017 season, sentinel plots were established in 24 states across the eastern US. The sentinel plots were scouted weekly until disease was observed. DM disease outbreaks were reported to the cucurbit ipmPIPE website (http://cdm.ipmpipe.org/) in 150 counties in 2017. Disease forecasting was conducted 2-3 times a week during the growing season from March to October, 2017. This information was available at the cucurbit ipmPIPE website. Regional reports of cucurbit DM occurrences to the cucurbit ipmPIPE website was kept up to date by the PD and co-PIs, ensuring accurate information was available to growers during 2017. Webinars, open houses, field days and trial tours also occurred in 2017 to alert and inform growers, industry and extension personnel of results of 2017 research (summarized in "How have the results been disseminated to communities of interest"). Results of 2017 trials testing cultivars for DM resistance allows evaluation of the performance of these cultivars over geographical areas and seasonal weather differences, generating valuable information for growers. Another year of fungicide efficacy trials provided growers recommendations for managing downy mildew that are up to date and tailored to their regions. Using social media to communicate results during the 2017 growing season as they occurred ensured that growers received the latest DM information and recommendations in a timely manner. Objective 6. Attract students to agricultural sciences. Robert Pyne completed his Ph.D. (Dissertation: Introgression of genetic resistance to downy mildew (Peronospora belbahrii) in a non-model plant species, sweet basil (Ocimum basilicum) in 2017 under the supervision of co-PIs Simon and Wyenandt. Many of the undergraduate students that have participated in the summer research program have moved on to productive careers/graduate programs in agricultural sciences: Andrew Aldcroft (Incotec Integrated Coat & Seed), Jesse Yamagata (Ph.D. Program in Plant Pathology, NCSU), Aidan Shands (Ph.D. program in Plant Pathology, University of California Riverside), Noah Luecke (Ph.D. program in Plant Science, University of Houston), Philip Engelgau (Ph.D. program in Horticulture, Michigan State University), Samuel Cude (USDA Agricultural Research Service), Juan Cerda (Ph.D. program in Bioinformatics, Pennsylvania State University), and Chelsea Newbold (currently applying to various Plant Pathology programs). Their work has been presented at the American Phytopathological Society Annual Meeting, the Oomycete Molecular Genetics Network Meeting, the Kelmen Scholars program at NCSU, and at local research events with CSUMB's Undergraduate Research Opportunity Center

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Chitwood, J., Shi, A., Mou, B., Evans, M., Clark, J., Motes, D., Chen, P., and Hensley, D. 2016. Population structure and association analysis of bolting, plant height, and leaf erectness in spinach. HortScience 51:481-486
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Koroch, A.R., Simon, J.E., and Juliani, H.R. 2017. Essential oil composition of purple basils, their reverted green varieties (Ocimum basilicum) and their associated biological activity. Industrial Crops and Products 107:526-530. https://doi.org/10.1016/j.indcrop.2017.04.066
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lange, H.W., Smart, C.D., and Seaman, A.J. 2016. Evaluation of fungicides allowed for organic production on downy mildew of cucumber, 2015. Plant Disease Management Reports 10:V015. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lee, R., Simon, J.E., Reichert, W., Juliani, R., and Tepper, B.J. 2017. Development of a lexicon for fresh basil aroma with links to volatile chemical composition. Perfumer & Flavorist 42:37-40,42-50.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ma, J., Shi, A., Mou, B., Evans, M., Clark, J., Motes, D., Correll, J., Xiong, H., Qin, J., Chitwood, J., and Weng, Y. 2016. Association mapping of leaf traits in spinach (Spinacia oleracea L.). Plant Breeding 135:399-404. doi:10.1111/pbr.12369
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Pyne, R., Honig, J., Vaiciunas, J., Bonos, S., Wyenandt, A., and Simon, J.E. 2017. A first linkage map and downy mildew resistance QTL discovery for sweet basil (Ocimum basilicum) facilitated by double digestion restriction site associated DNA sequencing (ddRADseq). PLoS ONE 12(9):e0184319.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Rahman, A., Miles, T.D., Martin, F.N., and Quesada-Ocampo, L.M. 2017. Molecular approaches for biosurveillance of the cucurbit downy mildew pathogen, Pseudoperonospora cubensis. Canadian Journal of Plant Pathology 3:282-296. DOI: 10.1080 /07060661.2017.1357661.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N. 2017. Evaluation of low-risk fungicides and host plant resistance for control of basil downy mildew, 2016. Plant Disease Management Reports 11:V045. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N. 2017. Evaluation of fungicides for control of downy mildew on cucumbers, 2016. Plant Disease Management Reports 11:V147. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N. 2017. Evaluation of fungicide programs for control of downy mildew on crisphead lettuce, Spring 2017. Plant Disease Management Reports 11:V153. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N., Hartman, D., and Vital, J. 2017. Evaluation of SAR compounds alone and in combination with a protectant for control of lettuce downy mildew, Spring 2017. Plant Disease Management Reports 11:V154. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N., Hartman, D., and Vital, J. 2017. Evaluation of fungicides for control of downy mildew on crisphead lettuce, Spring 2017. Plant Disease Management Reports 11:V155. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N., Vital, J., and Hartman, D. 2017. Evaluation of fungicide drench treatments for control of basil downy mildew, 2016. Plant Disease Management Reports 11:V031. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Shirley, A.M., Vallad, G.E., Kalb, S., Hughes, B., and Willis, R. 2017. Evaluation of selected fungicides for management of downy mildew on cucumber, fall 2016. Plant Disease Management Reports 11:V060. Online.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Thomas, A., Carbone, I., Choe, K., Quesada-Ocampo, L.M., and Ojiambo, P. 2017. Resurgence of cucurbit downy mildew in the United States: Insights from comparative genomic analysis of Pseudoperonospora cubensis. Ecology and Evolution 7:6231-6246. DOI: 10.1002/ece3.3194
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wallace, E.C., and Quesada-Ocampo, LM. 2017. Analysis of microsatellites from the transcriptome of downy mildew pathogens and their application for characterization of Pseudoperonospora populations. PeerJ 5:e3266 https://doi.org/10.7717/peerj.3266.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Wyenandt, C.A., Kline, W.L., and Ward, D.L. 2017. Effect of fungicide program on the development of downy mildew in three cucurbit crops in New Jersey. Plant Health Progress (https://doi.org/10.1094/PHP-04-17-0026-PHM)
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Xu, C., Jiao, C., Sun, H., Cai, X., Wang, X., Ge, C., Zheng, Y., Liu, W., Sun, X., Xu, Y., Deng, J., Zhang, Z., Huang, S., Dai, S., Mou, B., Wang, Q., Fei, Z., and Wang, Q. 2017. Draft genome of spinach and transcriptome diversity of 120 Spinacia accessions. Nature Communications 8:15275. doi: 10.1038/ncomms15275
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Crandall, S.G., Rahman, A., Quesada-Ocampo, L.M., Martin, F.N., Bilodeau, G.J., and Miles, T.D. 2017. Advances in diagnostics of downy mildews: lessons learned from other oomycetes and future challenges. Plant Disease Feature Article (First Look).
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Wyenandt, C.A., Maimone L.R., Homa, K., Madeiras, A., Wick, R.L., and Simon, J.E. 2017. Detection of downy mildew (Peronospora belbahrii) in seed of basil (Ocimum spp.) following field epidemics in southern New Jersey. HortTechnology
  • Type: Journal Articles Status: Accepted Year Published: 2018 Citation: Wyenandt, C.A., McGrath, M.T., Everts, K.E., Rideout, S.L., Gugino, B.K., and Kleczewski, N.M. 2018. Fungicide resistance management guidelines for cucurbit downy and powdery mildew control in the mid-Atlantic and Northeast regions of the United States in 2018. Plant Health Progress
  • Type: Theses/Dissertations Status: Published Year Published: 2017 Citation: Pyne, Robert. 2017. Ph.D. Plant Biology, School of Environmental and Biological Sciences. Introgression of Genetic Resistance to Downy Mildew (Peronospora belbahrii) in a Non-Model Plant Species, Sweet Basil (Ocimum basilicum). 193 pp. Rutgers, The State University of New Jersey.
  • Type: Other Status: Other Year Published: 2017 Citation: DArcangelo, K., Miles, T., and Quesada-Ocampo, L.M. 2017. Occurrence of fungicide resistance in Pseudoperonospora cubensis populations causing cucurbit downy mildew in commercial and wild hosts. Abstract. American Phytopathological Society Annual Meeting, San Antonio, TX, Aug.
  • Type: Other Status: Other Year Published: 2017 Citation: Mandal, M.K., Ikerd, J., Wallace, E., Turecheck, W., Grace, R., Quesada-Ocampo, L.M., and Kousik, C.S. 2017. Population biology of the downy mildew pathogen on tolerant and susceptible cucumber in the Southeastern United States. Abstract. American Phytopathological Society Annual Meeting, San Antonio, TX, Aug.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M. 2017. Downy mildew found on cucumber in southeast Michigan, homeowners should be on alert for downy mildew. Michigan State University Extension News for AgricultureVegetables: 3 Jul. Online.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M. 2017. Downy mildew confirmed on cucumber in Michigan and Ohio, growers should scout fields and protect their crops from downy mildew, now confirmed in Michigan in 2017. Michigan State University Extension News for AgricultureVegetables: 29 Jun. Online.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M. 2017. Ontario reports downy mildew on cucumber and cantaloupe, Michigan cucurbit growers should protect their crops from downy mildew. Michigan State University Extension News for AgricultureVegetables: 28 Jun. Online.
  • Type: Other Status: Other Year Published: 2017 Citation: Shands, A., Wallace, E., Miles, T., and Quesada-Ocampo, L.M. 2017. Detection of fungicide resistant Pseudoperonospora cubensis isolates using novel molecular tools. Oomycete Molecular Genetics Network Meeting, Pacific Grove, CA, Mar.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M.K., and Goldenhar, K. 2017. Downy mildew recommendations for cucumbers. Michigan State University Extension News for AgricultureVegetables: 14 Jun. Online.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M. 2017. Downy mildew found on cucumber in southeast Michigan, homeowners should be on the alert for downy mildew in their garden. Michigan State University Extension News for AgricultureHome Gardening/Vegetable Gardening: 3 Jul. Online.
  • Type: Other Status: Published Year Published: 2017 Citation: Hausbeck, M.K., and Linderman, S.D. 2017. Monitoring and managing cucurbit downy mildew. Online at https://veggies.msu.edu/wp-content/uploads/2017/05/FS_Cucurbit_DM_2017.pdf.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M.K., Morrice, J., and Linderman, S.D. 2016. Management of cucurbit downy mildew for gardeners. Online at https://veggies.msu.edu/extension-publications/#FactSheets.
  • Type: Other Status: Published Year Published: 2017 Citation: Quesada-Ocampo, L., and DArcangelo, K. 2017. Cucumber downy mildew confirmed in North Carolina and South Carolina. NC State Extension Plant Pathology Newsletter: 6 Jun. Online
  • Type: Other Status: Published Year Published: 2017 Citation: Smart, C.D. 2017. Cucurbit downy mildew management. VegEdge Newsletter: Feb.


Progress 03/01/16 to 02/28/17

Outputs
Target Audience:Our target audience includes people in the scientific, extension and agricultural communities. Included are other scientists, undergraduate and graduate students and post-docs in plant sciences, plant breeding and plant pathology, crop consultants, extension agents, crop protection industry, growers, shippers, processors and other allied stakeholder industries. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has been training many people in many field- and laboratory-based agricultural scientific techniques such as spore trapping; weather monitoring; field plot design, maintenance and disease evaluation; diagnostics; growth and storage of DM pathogens; development of diagnostic tools; genomics; bioinformatics; biomathematics; DNA extractions; PCR; qPCR; SSR/microsatellite genotyping; Sanger sequencing; statistical analyses. People who have benefited from this training include post-docs (Alamgir Rahman, Surendran Arumugam), graduate students (Andy Shirley, Katelyn Goldenhar, Kim D'Arcangelo, Lauren Brzozowsi, Marth Suderman, Maureiq Ojwang, Trevor Ruiz, Robert Pyne, Kathryn Homa), undergraduate students (Aidan Shands, Andrew Aldcroft, Gina Dabbah, Jesse Yamagata, Phi Diep, Philip Engelgeau, Thilani Jayakody, Amber Glenning, Courtney Lepping, Jordan Castellari, Lisa Panichelli, Tanner Boss), and technicians (Brian Sheppard, Chris DelCastillo, Emi Kuroiwa, Evan McCllenthan, Marina Ramon, Paige Reeves, Rachel Kreis, Scott Hughes, Sheila Linderman, Steve Kalb). Aidan Shands, Andrew Aldcroft, and Jesse Yamagata have participated in the summer research program for undergraduate students, one of the objectives of this project. How have the results been disseminated to communities of interest?Research results have been disseminated via demonstration plots, publications, websites, scientific/extension presentations, webinars, and scientific/extension meetings. Scientific Meetings: Presentations to the scientific community acknowledging NIFA included 4 talks and 2 posters at the XIth Eucarpia Cucurbitaceae Meeting, Warsaw, Poland, Jul; the Canadian Phytopathological Society Annual Meeting, Biovigilance Symposium, Moncton, Canada, Jun; the American Phytopathological Society Meeting (1 talk and 2 posters), Tampa, FL, Aug; the Department of Plant, Soil, and Microbial Sciences Seminar, Michigan State University, East Lansing, MI, Jun. Other scientific presentations included talks at the Department of Biochemistry Seminar, North Carolina State University, Raleigh, NC, Feb; the National Association of Plant Breeders (NAPB) National Meeting, Raleigh, NC, Aug; and posters at the 7th Annual Conference of the American Council for Medicinally Active Plants (ACMAP), Lima, Peru, Jun-Jul; and the northeastern Plant, Pest and Soil Conference, Philadelphia, PA, Jan. Three undergraduate researchers presented posters at the NCSU Undergraduate Research Symposium, Raleigh, NC, Aug; two of these undergraduates participated in this grant's summer research program. Extension Meetings: Research results were presented at 23 meetings at the local, regional and national levels, and as 5 webinars, to growers, commodity groups, chemical companies, extension educators, and other agricultural industry stakeholders. Meetings included the 31st Annual Southeast Vegetable and Fruit Expo, Myrtle Beach, NC; Southeast Regional Fruit and Vegetable Conference, Savannah, GA; and the Great Lakes Fruit, Vegetable and Farm Market Expo (2 talks), Grand Rapids, MI; Pickle Packers International Annual Meeting, Charleston, SC; Pickle Packers International Spring Meeting, Raleigh, NC; Empire State Producers Expo, Syracuse, NY; Seminis Meeting, Charleston, SC; Small Farm Conference, Danville, IN; Valent Meeting, East Lansing, MI; North Carolina Watermelon Convention, Wrightsville Beach, NC; NJ Twilight Meeting, Snyder Research and Extension Farm, Pittstown, NJ; AgBiome Meeting, Durham, NC; Discovering Geneva Community Event, Geneva, NY; North Country Willsboro Research Farm Summer Growers Meeting, Willsboro, NY; Crop Consultant Meeting, Syracuse, NY; Canton Summer Growers Meeting, Canton, NY; New Jersey Vegetable Growers Association Annual Agricultural Convention, Atlantic City, NJ; Northeast Organic Farming Association-New Jersey Annual Winter Conference, New Brunswick, NJ; Lettuce Advisory Committee Meeting, Everglades Research and Education Center, Belle Glade, FL; Leadership Glades Meeting, Everglades Research and Education Center, Belle Glade, FL; Plant Pathology Teacher Workshop, Everglades Research and Education Center, Belle Glade, FL. Field Days, Tours, Workshops: Research was featured at 11 extension field days, tours, and workshops: Michigan State University Plant Pathology Farm Tour, Lansing, MI; Western New York Field Days, Portland, NY; Cucurbit Field Day, Cleveland, NC; Agent Training Workshop, Extension Conference, Raleigh, NC; Downy Mildew Experiment Tour, Hubbard, OR; Twilight Tour, Rutgers Agricultural Research and Extension Center (RAREC), Bridgeton, NJ; Downy Mildew Field Trials and Sentinel Plots Tour, Florida Ag UF/IFAS Gulf Coast Research and Education Center, Wimauma, FL; Basil Variety Trial Tour, Florida Ag UF/IFAS Gulf Coast Research and Education Center, Wimauma, FL; Cornell Vegetable Breeding Institute Annual Field Days, Homer Thompson Vegetable Research Farm, Freeville, NY; Spinach Downy Mildew Breeding Plots Tour, California Leafy Greens Research Program, USDA-ARS Station, Salinas, CA; Cucurbit and Basil Downy Mildews Fungicide Efficacy Trials, Twilight Tour, Rutgers Agricultural Research and Extension Center, Bridgeton, NJ. What do you plan to do during the next reporting period to accomplish the goals?OBJECTIVE 1. Early detection and rapid diagnostics: research will occur in two areas. 1i) Species-specific genetic markers for diagnostics: Martin will continue developing mitochondrial diagnostic markers will continue to be developed and provide sequencing support; Quesada will continue identifying nuclear markers for P. cubensis and provide sequencing data; Gent and Smart will continue developing mitochondrial SNP assays. 1iii) Fungicide-resistance diagnostics: Martin will begin developing assays for detection of fungicide resistance; Gent, Hausbeck, Quesada, Smart, and Vallad will begin testing markers and provide isolates; Miles will begin developing RPA assays for detection of fungicide resistance; Goldy, Ojiambo and Raid will provide isolates. OBJECTIVE 2. Tracking pathogen dispersal and survival: activities will occur in four areas. 2i) Expand the national DM tracking site: Ojiambo will continue ipmPIPE administration, in particular developing and testing the basil DM website; and Hausbeck, Quesada, Smart, Smith, Vallad and Wyenandt will continue sentinel plot deployment. 2iii) Provide grower alerts based on DM inoculum strength: Ojiambo will continue data collection and modeling; Bhattacharyya and Gent will continue modeling; Hausbeck, Quesada, Smart, Vallad and Wyenandt will continue data collection. 2iv) Determine DM survival: Hausbeck, Quesada, Ojiambo and Vallad will continue research. 2v) Quantify duration of disease control with DM fungicides: Ojiambo will continue field experiments and modeling; Hausbeck, Smart, Vallad and Wyenandt will continue field experiments. OBJECTIVE 3. Developing DM-resistant crops: research will occur in two areas. 3i) Develope DM-resistant crop germplasm: Mazourek, Mou and Simon will continue research. In particular, Mazourek will continue pedigree breeding approaches. 3ii) Participatory plant breeding of DM-resistant cucumbers: Agehara, Goldy, Hausbeck, Mazourek, Quesada, Smart and Smith begin this research, Agehara will make selections with grower input, and Mazourek will expand multilocation trials if sufficient seed is available. OBJECTIVE 4. Cost/benefit ratios and grower adoption: research will occur in two areas. 4i) Baseline DM disease impacts to food security: Govindasamy will continue to determine impacts with input from Hausbeck, Quesada, Raid, Smart, Smith, Vallad and Wyenandt. 4ii) Economic impact of the behavioral, social, environmental outcomes: Govindasamy will continue evaluation of impacts with the aid of Hausbeck, Quesada, Raid, Smart, Smith, Vallad and Wyenandt. OBJECTIVE 5. Outreach and adoption: activities will occur in four areas. 5i) DM monitoring/forecasting system: Ojiambo will continue providing details on monitoring/forecasting system; and Agehara, Hausbeck, Quesada, Raid, Smart, Smith, Vallad, and Wyenandt will continue reporting pathogen outbreaks, provide educational materials and training for growers, industry and extension personnel during year 2. 5ii) Deployment of DM resistant breeding lines/varieties: Hausbeck, Quesada, Raid, Smart, Vallad and Wyenandt will evaluate host resistance over a wide geographic area; and Agehara, Goldy and Smith will evaluate horticultural traits. 5iii) Seed testing for basil DM: Hausbeck, Quesada, Vallad and Wyenandt will start conducting seed testing experiments; and Agehara, Goldy, Hausbeck, Quesada, Raid, Smart, Vallad and Wyenandt will begin to develop/disseminate educational materials on the importance of clean seed. 5iv) Field spray recommendations: Hausbeck, Quesada, Raid, Smart, Vallad and Wyenandt will continue providing input on fungicide success or failures seen in their region, and extending these best management practices to growers, extension and industry personnel. 5vi) Develop/frequently update social media: Agehara, Goldy, Hausbeck, Quesada, Raid, Smart, Smith, Vallad and Wyenandt will continue developing social media and posting frequent updates. OBJECTIVE 6. Attract students to ag sciences: the PD and all PIs will continue to mentor post-docs, graduate and undergraduate students who participate in project research. The summer research program for undergrads will continue to be implemented.

Impacts
What was accomplished under these goals? OBJ 1. Assembled mitochondrial genomes of 328 isolates representing 7 genera and 18 species provide additional support for marker development. Two different marker systems have been developed for P. cubensis: a rapid gel-based assay based on amplicon size to differentiate P. cubensis from related taxa, and a TaqMan real-time PCR assay. The TaqMan assay will amplify both P. cubensis and P. humuli with species-specific TaqMan probes for detection of each species. Specificity of the markers was evaluated and observed detection of P. cubensis was highly specific; the P. humuli assay is in the final stages of development/validation. Mitochondrial haplotype of the 12 P. cubensis and 16 P. humuli mitochondrial genomes were analyzed to support population studies. OBJ 1. Genomic regions unique to and conserved in P. cubensis isolates were identified through bioinformatics, and validated using PCR against a larger collection of isolates of P. cubensis, P. humuli and other oomycetes. Seven diagnostic markers were found to be specific to P. cubensis. These markers are being used for pathogen diagnostics on infected tissue and some are being adapted for monitoring airborne inoculum with real-time PCR and spore traps. For early detection using spore traps with species-specific markers, genomic marker c3155.4e9 significantly improved sensitivity of P. cubensis DNA detection on leaf disks (LD), sampling rods (ROD), and sporangia suspension (SPR) using probe-based qPCR. OBJ 1. High resolution melt analysis (HRM) correctly diagnosed all tested isolates of P. cubensis and P. humuli as well as symptomatic plants collected in the field. The LNA and HRM assays correctly identified both species when tested independently in a second laboratory. A nested conventional PCR was accurate for 13 of 15 P. cubensis isolates, with two isolates collected from squash hosts inaccurately identified as P. humuli due to non-conserved SNPs lying within the reverse primer sequences. OBJ 2. Discussions were also held to determine the long-term hosting of the website. The goal of expanding the national DM tracking site is in progress and on track to be met. OBJ 2. Hausbeck confirmed cucurbit DM only on cucumber 'Straight Eight' in sentinel plots on 9 Aug and only in Ingham County, MI. Quesada confirmed first detection of DM occurred 6 Sep. Smart confirmed first symptoms of DM appeared in late Jul in Geneva, NY. Smith conducted one DM evaluation in Watsonville, CA; no mildew developed in this trial. Vallad initially confirmed DM in FL on 17 Oct. OBJ 2. MI cucurbit DM outbreaks were reported to the Cucurbit ipmPIPE website and continually uploaded to www.veggies.msu.edu, www.downymildew.msu.edu during the growing season. NC cucurbit DM outbreaks were reported to the CDM ipmPIPE and disease alerts for other DM were disseminated using Extension websites and Twitter. NY cucurbit DM outbreaks were reported to the CDM ipmPIPE and disease alerts were disseminated using Extension websites and Twitter. FL sentinel plots were established in early September, but DM was not confirmed until Oct 17. OBJ 2. Development of a network model is in progress. Analyses were completed that related the size of the initial disease focus and the final extent of cucurbit DM, and quantified the consistency of models that predict the velocity of epidemic expansion. A significant interaction was observed of the spread parameter of a power law dispersal model for epidemic years where data were well described by the power law model, suggesting that the spread parameter may not be stable over multiple epidemic years. However, value of ≈ 2 of the parameter may be considered the lower limit of the distance traveled by epidemic wave-fronts for aerially transmitted pathogens that follow a power law dispersal function. Analysis of historical data from the CDM ipmPIPE data found evidence of a relationship between the initial area affected by cucurbit DM along the Gulf coast and final extent of disease across the eastern US data sets. There was a significant correlation relationship between current and final epidemic area as early as late April, well before cucurbit crops in northern latitudes are even planted. These findings suggest that the magnitude of epidemics at the landscape level may be largely determined by antecedents associated with overwintering success of the pathogen in southern latitudes. A manuscript detailing these findings was prepared and submitted. OBJ 2. Koch's postulates are being completed in the lab for isolates from wild or alternative host that presented a CDM infection based on visual inspection and a molecular confirmation of CDM as the cause of infections observed in the field. Molecular confirmation of a CDM infection is being performed by amplifying and sequencing the internal transcribed spacer (ITS) region. OBJ 2. Especially effective fungicides in MI testing were Omega, Orondis Opti, Orondis Ultra, Zing! and V-10208 applied alone and alternating programs of Orondis Opti, Presidio, Ranman, Zampro and Zing!, often tankmixed with Bravo WeatherStik. Smart found best treatments in NY as measured by AUDPC were Orondis Gold drench, Zing! and Ranman. Based on AUDPC treatments, Vallad found Orondis Opti A, Omega, Raman + Silwet L-77, Ranman + Bravo Weatherstik, Zampro, and Presidio provided the greatest level of disease control in FL. Conventional and organic fungicides and different fungicide programs were evaluated for the control of cucurbit DM and basil DM at RAREC, Bridgeton, NJ. OBJ 3. Cucumber: 2,684 plants of cucumber breeding populations were evaluated for disease severity, earliness and productivity. Cuttings were rooted from the best 57 individuals. These are being self-pollinated, backcrossed and/or intermated according to which pollination scheme was determined to be the most appropriate toward the development of uniform germplasm that combines all the appropriate characteristics. OBJ 3. A first generation genetic map was developed from a basil population segregating for DM resistance. One major and two minor QTLs associated with DM resistance were identified. Individuals in the F2 generation were used as a base population for DM resistance breeding to develop advanced breeding lines. OBJ 3. Basil host-pathogen RNAseq: Relative expression is being compared to identify resistance and susceptibility related genes in the host and pathogen. OBJ 4. Institutional review board (IRB) formalities and requirements have been met and the survey will be finalized after review by mid-Atlantic region panelists. The full survey will be conducted during Mar 2017. OBJ 5. Cucumber DM pathogen outbreaks were reported at the cucumber DM ipmPIPE website. PIs also communicated pathogen outbreaks via alerts published online at extension websites, phone calls, and in-person visits. OBJ 5. PIs have presented recommendations at extension meetings and via proceedings published online at meeting websites, alerts published online at extension websites, phone calls, and in-person visits. OBJ 5. PIs regularly used social media to update growers. OBJ 6. Two post-docs, 9 graduate students, and 12 undergraduate students are involved in the research of this project. See information in Section 5.b.i. Training for more details. OBJ 6. Three of the undergraduate students have participated in the summer research program. See information in Section 5.b.i. Training for more details.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adams, M. L., and Quesada-Ocampo, L. M. 2016. Evaluation of fungicides for control of downy mildew on cucumber, Cleveland 2015. Plant Disease Management Reports 10:V085.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adams, M. L., and Quesada-Ocampo, L. M. 2016. Evaluation of fungicides for control of downy mildew on cucumber, Kinston 2015. Plant Disease Management Reports 10:V086.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lange, H.W., Smart, C.D., and Seaman, A.J. 2016. Evaluation of fungicides allowed for organic production on downy mildew of cucumber, 2015. Plant Disease Management Report 10:V015.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Withers, S., Gongora-Castillo, E., Gent, D., Thomas, A., Ojiambo, P., and Quesada-Ocampo, L. M. 2016. Using next-generation sequencing to develop molecular diagnostics for Pseudoperonospora cubensis, the cucurbit downy mildew pathogen. Phytopathology 106: 1105-1116.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Naegele, R. P., Quesada-Ocampo, L. M., Kurjan, J. D, Saude, C., and Hausbeck, M. K. 2016. Regional and temporal population structure of Pseudoperonospora cubensis in Michigan and Ontario. Phytopathology 106:372-379.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adams, M. L., Noel, N. A., and Quesada-Ocampo, L. M. 2016. Evaluation of fungicides for control of downy mildew on cucumber, Clayton 2015. Plant Disease Management Reports 10:V084.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lange, H.W., Smart, C.D., and Seaman, A.J. 2017. Evaluation of materials allowed for organic production on downy mildew of cucumber, 2016. Plant Disease Management Report 11:V015.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N., Vital, J., and Hartman, D.A. 2017. Evaluation of fungicide drench treatments for control of basil downy mildew, 2016. Plant Disease Management Reports 11:V031.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Raid, R.N. 2017. Evaluation of low-risk fungicides and host-plant resistance for control of basil downy mildew, 2016. Plant Disease Management Reports 11:V045.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Raid, R.N., Hartman D.A., and Vital, J. 2016. Evaluation of sweet basil varieties for resistance to downy mildew, 2015. Plant Disease Management Reports 10:V115.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Raid, R.N., Moreira, D., and Vital, J. 2016. Evaluation of fungicide programs for control of lettuce downy mildew, Spring 2016. Plant Disease Management Reports 10:V131.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Raid, R.N., Moreira, D., and Vital, J. 2016. Evaluation of Torac for use in a management program for control of lettuce downy mildew, Spring 2016. Plant Disease Management Reports 10:V130.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Hausbeck, M.K., Brisco-McCann, E.I., Goldenhar, K.E., and Cook, A.J. 2017. Evaluation of fungicide programs for control of downy mildew of cucumber, 2016. Plant Disease Management Reports 11:V063.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Goldenhar, K.E., and Hausbeck, M.K. 2017. Evaluation of fungicides for control of downy mildew of cucumber when applied after pathogen establishment, 2016. Plant Disease Management Reports 11:V080.
  • Type: Other Status: Published Year Published: 2016 Citation: Bello, J., Guo Y., and Hausbeck, M. 2016. Population structure of Pseudoperonospora cubensis isolates in Michigan and Ontario, Canada. Abstract. Phytopathology 106 (Suppl.):S4.140.
  • Type: Other Status: Published Year Published: 2016 Citation: Wallace, E., and Quesada-Ocampo, L. 2016. Pseudoperonospora cubensis on commercial and non-commercial cucurbits in North Carolina: population structure determined by simple sequence repeats (SSRs). Abstract. Phytopathology 106 (Suppl.):S4.12.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M. 2016. Early infection of downy mildew found in Michigan cucumbers. Michigan State University Extension News for AgricultureVegetables: 11 Jul. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M. 2016. Downy mildew confirmed on cucumbers in Ohio. Michigan State University Extension News for AgricultureVegetables: 7 Jul. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M. 2016. Downy mildew on cucumber reported in Ontario. Michigan State University Extension News for AgricultureVegetables: 6 Jul. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M. 2016. Downy mildew spends a decade damaging cucumbers. Vegetable Grower News 50(5):16-17.
  • Type: Other Status: Published Year Published: 2016 Citation: Hausbeck, M.K., and Goldenhar, K. 2016. Downy mildew prevention and control. Pages 9-12 in: Pickling Cucumber Session Summaries, Great Lakes Fruit, Vegetable and Farm Market Expo, Grand Rapids, MI, Dec. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Smart, C., and Lange, H. 2016. Cucurbit downy mildew update. Article for the VegEdge 12(2):4. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Smart, C., and Lange, H. 2016. Vine crop update 2015. Proceedings of the 2016 Empire State Producers Expo, Syracuse, NY.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Cucurbit downy mildew update: 8/17/16. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition, 17 Aug. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Cucurbit downy mildew update confirmed on cucumber in north Jersey 7-27-16. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition, 27 Jul. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Cucurbit downy mildew update confirmed on cucumber in southern New Jersey  ALERT  7/11/16. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition, 11 Jul. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Cucurbit downy mildew update confirmed on cucumber in Maryland  ALERT. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition, 29 Jun. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Basil downy mildew confirmed in southern and central New Jersey  ALERT 6/30/16. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition/Organic Farm Advisory, 30 Jun. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Basil downy mildew  24cs granted for Subdue Maxx and Heritage use in greenhouse. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition/Commercial Ag Updates + Farm Food Safety, 16 May. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Wyenandt, A. 2016. Basil downy mildew workshop session videos. Rutgers University Cooperative Extension Plant & Pest Advisory, Vegetable Crops Edition, 7 Mar. Online.
  • Type: Other Status: Published Year Published: 2016 Citation: Quesada-Ocampo, L. M. 2016. Watermelon downy mildew reported in North Carolina. Extension Plant Pathology Portal. 17 Jun.
  • Type: Other Status: Published Year Published: 2016 Citation: Quesada-Ocampo, L. M. 2016. Cucumber downy mildew reported in North Carolina. Extension Plant Pathology Portal. 1 Jun.