PROPOSAL COVER PAGE Sponsor: Hatch-Individual Hatch-Multidisciplinary McIntire-Stennis-Individual McIntire-Stennis Multidisciplinary Project Title: Pratylenchus penetrans as a pest of soybean Will this proposal have an integrated/extension component? Is this proposal related to an official Multistate project? YES NO YES NO Is this a revision of a previous proposal? If yes, which Multistate project number? YES NO NC1197 Which USDA National Priorities does this proposal link to? Global Food Security: Crops and Agronomic Plants Global Food Security: Livestock and Poultry Climate Change and Energy Needs Sustainable Use of Natural Resources Nutrition Food Safety Education and Science Literacy Rural Prosperity Which USDA Knowledge Area(s) does this proposal link to? Insert Code(s) 212 Project Period: 10/01/2017 to 09/30/18 09/30/19 09/30/20 09/30/21 Individual Name (lastname, firstname) 9 month or 12 month appointment Principal Investigator: MacGuidwin, Ann 9 month 12 month Department Plant Pathology Individual Name (lastname, firstname) 9 month or 12 month appointment Co-PI(s): 9 month 12 month 9 month 12 month 9 month 12 month Summary Budget Request: Year 1 Year 2 Year 3 Year 4 Research Assistant Salary: 22,081 22,081 0 0 Student Hourly/LTE Salary: 1,540 462 0 0 Total RA and Student HourlyWages: 23,621 22,543 0 0 Fringe Benefits: 5,314 5,288 0 0 Supplies: 4,000 1,000 0 0 Travel: 500 100 0 0 Total Request: 33,435 28,931 0 0 Summary of Proposed Work: Total Award Budget: 62,366 The genus Pratylenchus is the most common nematode pest of soybean in Wisconsin. Three of the five species shown to damage soybean under controlled conditions have been reported here. One, P. penetrans, is known to cause yield loss for other crops in Wisconsin so our first objective is to develop a model to estimate the percentage of soybean yield lost as a function of P. penetrans population density. This will be the first damage function model of the P. penetrans/soybean interaction based on field-scale data. We will use the results to raise awareness about nematode pests and to improve feedback by our diagnostic service. The prevalence of different species of Pratylenchus is not known for Wisconsin so objective two is to identify bisexual species to test the hypothesis that P. penetrans is the prevalent bisexual species in Wisconsin. About half of the 89 currently recognized Pratylenchus species are bisexual (comprised of males and females) as are four of the five species with demonstrated pathogenicity to soybean. Eight bisexual species are on record for Wisconsin so this research is important for assessing the frequency and distribution of known soybean pests and presents opportunity to discover species new to Wisconsin or science. We observed in previous work that P. penetrans males do less damage to roots than females so our third objective is to characterize gender differences in feeding activities and host tissue response. This research will be useful for future studies aimed at identifying parasitism genes and their expression in P. penetrans. Reviewer Names Please provide names and contact information (department and email address) for at least three independent reviewers you believe are especially well qualified to review this proposal. They should have no conflicts of interest with the proposal PIs. A conflict of interest constitutes anyone with whom the applicant has collaborated or published research in the previous three year period. You may also provide names of reviewers whom you would prefer not to review this proposal and indicate why. Reviewer Name/Department Email Address Patty McManus Plant Pathology [email protected] Russ Groves Entomology [email protected] Glen Stanosz Forest and Wildlife Ecology [email protected] WORKSHEET FOR PROJECT BUDGET Detailed Budget for Year: 1 Project Title: Pratylenchus penetrans as a pest of soybean Project Period: 10/01/2017 -- 09/30/2018 Individual Name (lastname, firstname) % Effort on Project (Will be Payrolled Effort) Principal Investigator: MacGuidwin, Ann 10% Co-PI(s): Salaries/Wages & Fringe Benefits: Salary/ Individual Name Wage Salaries/ Fringe Budget Category (lastname, firstname) Base Months Effort Wages Rate (%) Fringes Research Assistant: Kanan Kutsuwa 44,162 12 50% 22,081 23.9% 5,277 (annual base) (# months) (appt %) Student Hourly/LTE: TBD 11.00 7 20 1,540 2.4% 37 (hourly rate) (hrs/week) (# weeks) Subtotal 23,621 5,314 Supplies & Travel: Supplies (provide detail below*): 4,000 Travel: 500 Experimental Animals: Subtotal: 500 Page Charges (direct result of Hatch project only): Survey Costs (i.e., printing, postage) Subtotal: 4,000 Year 1 Total Budget Request: 33,435 *Details regarding Supplies and Travel [include one-time computer/printer requests/justification]: Growth chamber rental (obj 1) = $700 per experiment x 2 experiments = $1,400; supplies for nematode assays (obj. 1) = $500; supplies for in vitro cultures for inoculum and studies (obj 3) = $500; field supplies (obj 1) = $300; supplies for molecular identification (obj. 2) including DNA extraction kits, primers, chemicals, sequencing costs, etc. = $1,300. Travel = vehicle rental to travel to field plots WORKSHEET FOR PROJECT BUDGET Detailed Budget for Year: 2 Project Title: Pratylenchus penetrans as a pest of soybean Project Period: 10/01/2018 -- 09/30/2019 Individual Name (lastname, firstname) % Effort on Project (Will be Payrolled Effort) Principal Investigator: MacGuidwin, Ann 10% Co-PI(s): Salaries/Wages & Fringe Benefits: Salary/ Individual Name Wage Salaries/ Fringe Budget Category (lastname, firstname) Base Months Effort Wages Rate (%) Fringes Research Assistant: Kanan Kutsuwa 44,162 12 50% 22,081 23.9% 5,277 (annual base) (# months) (appt %) Student Hourly/LTE: TBD 11.00 7 6 462 2.4% 11 (hourly rate) (hrs/week) (# weeks) Subtotal 22,543 5,288 Supplies & Travel: Supplies (provide detail below*): 1,000 Travel: 100 Experimental Animals: Subtotal: 100 Page Charges (direct result of Hatch project only): Survey Costs (i.e., printing, postage) Subtotal: 1,000 Year 2 Total Budget Request: 28,931 *Details regarding supplies and travel [include one-time computer/printer requests/justification]: General lab supplies (nematode culture, assays) = $300, supplies and sequencing services (obj 2) = $700 and travel to field plots for sampling and harvest 2017 2018 2019 2020 2021 Major Activities Qtr 1 Qtr 2 Qtr 3 Qtr 4 Qtr 1 Qtr 2 Qtr 3 Qtr 4 Qtr 1 Qtr 2 Qtr 3 Qtr 4 Qtr 1 Qtr 2 Qtr 3 Qtr 4 Qtr 1 Qtr 2 Qtr 3 Qtr 4 coursework, prepare for and take prelim X X X finish field 2 season (obj 1) X develop model (obj 1) X X X X field season 3 (obj 1) X X X validate and modify model (obj 1) X X growth chamber dose/response experiments (obj 1) X X X X X morphologcial analysis and culture initiation (obj 2) X X X X X molecular identification (obj 2) X X X X X in vitro histopathology experiments (obj 3) X X X X write thesis X X defend thesis / graduate X Pratylenchus penetrans as a Pest of Soybean in Wisconsin Summary The genus Pratylenchus is the most common nematode pest of soybean in Wisconsin. Three of the five species shown to damage soybean under controlled conditions have been reported here. One, P. penetrans, is known to cause yield loss for other crops in Wisconsin so our first objective is to develop a model to estimate the percentage of soybean yield lost as a function of P. penetrans population density. This will be the first damage function model of the P. penetrans/soybean interaction based on field-scale data. We will use the results to raise awareness about nematode pests and to improve feedback by our diagnostic service. The prevalence of different species of Pratylenchus is not known for Wisconsin so objective two is to identify bisexual species to test the hypothesis that P. penetrans is the prevalent bisexual species in Wisconsin. About half of the 89 currently recognized Pratylenchus species are bisexual (comprised of males and females) as are four of the five species with demonstrated pathogenicity to soybean. Eight bisexual species are on record for Wisconsin so this research is important for assessing the frequency and distribution of known soybean pests and presents opportunity to discover species new to Wisconsin or science. We made observations in previous work that P. penetrans males do less damage to roots than females so our third objective is to characterize gender differences in feeding activities and host tissue response to male and female nematodes. This research will be useful for future studies aimed at identifying parasitism genes in P. penetrans. 1 Results from Previous Hatch Support: Current Individual Hatch (2012-2016): Practical Management of Nematodes on Corn, Soybean, and Other Crops of Regional Importance: Pathogenicity of Pratylenchus penetrans to Corn Toby Lunt completed his M.S. degree in Agroecology and Plant Pathology in 2014 and his M. S. project, “Investigating relationships between Pratylenchus penetrans and Fusarium verticillioides on corn seedlings” contributed important background to the Ph.D project of Tomas Rush, who joined the lab in 2014 and was funded by Hatch in 2015 to 2016. Toby found no evidence of a synergistic interaction for the nematode and fungus and his data indicated that the association might actually be detrimental to the nematode. Toby’s results were not conclusive enough to publish, but will be incorporated in papers written by Tomas Rush. Tomas recently completed an experiment with strong data confirming that the presence of the nematode does not alter corn seedling disease initiated from F. verticilliodes-infected seed. Like Toby, Tomas found the relationship between F. verticilliodes and P. penetrans was antagonistic to the nematode. Corn producers are anxious to know if nematicidal seed treatments should be considered in fields at risk for F.