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FY18 USWBSI INDIVDIUAL PROJECT PROPOSAL(S) NOTE: This document will be retained in the USWBSI’s NFO

COVER PAGE

Instructions: Update any contact information that is not correct and sign cover page before submitting full Individual Project Proposal(s).

USWBSI Consolidate Title for Agreement:

Diagnostic Services for DON

Principal Investigator (PI): Yanhong Dong PI’s Institution: University of Minnesota

PI’s Address: Department of Plant Pathology 495 Borlaug Hal 1991 Upper Buford Circle St. Paul, MN 55108

PI’s E-mail: [email protected] PI’s Phone: 612-625-2751

Fiscal Year (FY): 2018 Award Period: 8/1/18 – 7/31/19

ARS Agreement Number: New USWBSI’s FY18 Total

Recommended Amount: $ 274,009

USDA-ARS FY18 Total Award Amount: $ 265,513

USWBSI Project ID

USWBSI Research Category USWBSI Project Title

PI Requested Amount

USWBSI‘s Recommended

Amount

ARS Award

Amount

FY18-DO-002 FST Diagnostic Services for DON. $ 282,769 $ 274,009 $ 265,513

FY 18 USWBSI’s Total Recommended/Award Amount $ 274,009 $ 265,513

FY18 USWBSI INDIVIDUAL PROJECT PROPOSAL

FY18 USWBSI Individual Project Proposal

PROJECT SUMMARY PAGE Principal and Co- Investigator(s): Principal Investigator: Yanhong Dong Institution: University of Minnesota

Co-Investigator #1: Institution:



Project Title 1: Diagnostic Services for DON.

PROJECT SUMMARY

The goal of this project is to provide rapid, cost-effective and accurate mycotoxin analysis -

especially deoxynivalenol (DON) - for Fusarium Head Blight (FHB or scab) research projects.

The project will use gas chromatography-mass spectrometry (GC-MS) to provide quick and accurate

measurement for: 1) DON and related mycotoxins, such as 3-acetyl-DON, 15-acetyl-DON and

nivalenol (NIV); 2) Zearalenone for the projects approved by the executive committee; 3) mycotoxins

in mature wheat and barley seeds as well as individual kernel, spikelet, head, small leaf and stem

fragment at different disease development stages and fungal culture extract; 4) 35,485 and 36,285

DON samples for FY18 and FY19, respectively, from 42 FHB research groups and 22 states

including Arkansas, Delaware, Georgia, Idaho, Illinois, Indiana, Kansas, Kentucky, Louisiana,

Maryland, Michigan, Minnesota, Missouri, New York, North Carolina, North Dakota, Ohio,

Pennsylvania, South Dakota, Tennessee, Texas, and Wisconsin.

The DON data provided by the services is essential to breeding (traditional and molecular) projects

aiming at the development of wheat and barley varieties with improved resistance to the disease and

DON contamination. The services have been used for epidemiology, genetics and molecular studies

of the host, pathogen, and host-pathogen interaction aiming at improving our understanding of the

biology of the disease as well as developing effective disease control practices. The services have

also been used to assist the developments of other rapid DON screening methods such as IR and

Raman. The mycotoxin data provided to FHB researchers by the services gives researchers a means

to evaluate the effectiveness of their efforts in fighting Fusarium Head Blight.



PROJECT DESCRIPTION

Diagnostic Service for DON

Introduction:

The goal of this project is to provide rapid, cost-effective and accurate mycotoxin analysis - especially

deoxynivalenol (DON) - for Fusarium Head Blight (FHB or scab) research projects.

DON is one of the most common and abundant naturally occurring mycotoxins in wheat and barley, and is

produced predominately by Fusarium graminearum, the primary FHB pathogen in the USA (Leonard et al.,

2003). It adversely affects animals, especially swine by causing vomiting, refusal of feed and decreased

weight gain (D’Mello et al., 1999; Pestka, 2007). It may also cause vomiting in humans as well as

headache, fever, upset stomach, and diarrhea (Pestka et al., 2005; Pestka, 2010). Contamination of wheat

and barley with DON results in grains unacceptable for the marketplace, leading to huge economic losses to

the producers.

The project will use gas chromatography-mass spectrometry (GC-MS) to provide quick and accurate

measurement for: 1) DON and related mycotoxins, such as 3-acetyl-DON, 15-acetyl-DON and nivalenol

(NIV); 2) Zearalenone for the projects approved by the executive committee; 3) mycotoxins in mature

wheat and barley seeds as well as individual kernel, spikelet, head, small leaf and stem fragment at different

disease development stages and fungal culture extract; 4) 35,485 and 36,285 DON samples for FY18 and

FY19, respectively, from 42 FHB research groups and 22 states including Arkansas, Delaware, Georgia,

Idaho, Illinois, Indiana, Kansas, Kentucky, Louisiana, Maryland, Michigan, Minnesota, Missouri, New

York, North Carolina, North Dakota, Ohio, Pennsylvania, South Dakota, Tennessee, Texas, and Wisconsin.

Rationale and Significance:

Quick and reliable DON analysis services have been an important part of efforts to fight FHB. Monitoring

DON levels in wheat and barley lines from field experiments, nurseries and greenhouse screening has been

used to assists wheat and barley breeding programs to develop FHB resistant varieties (Smith et al., 2004;

Fuentes et al., 2005; Jiang et al., 2006, 2007; Yu et al., 2008, Costa et al., 2010; Lewis et al., 2010, 2012;

Bernardo et al., 2014; Clark et al., 2014; Van Sanford et al., 2016; Wang et al., 2017). Monitoring DON

levels is the only means to detect resistances that function as an ability to degrade or prevent the synthesis

of DON (Miller & Arnisen, 1986) and resistances that are observed as the plant tolerance to high DON

concentrations (Wang & Miller, 1988;) and to detoxify DON into a DON-glucose conjugate (Lemmens et

al., 2005). The ability to detect very low levels of DON in a single kernel, or spikelet has helped the study



of disease mechanisms. Levels of DON in individual spikelet during the early infection processes in barley

have been used to investigate when biosynthesis of DON occurs in plant (Evans et al., 2000), and to study

the relationship among disease development, DON accumulation, and host gene transcript accumulation

(Boddu et al., 2006). Establishing when toxins are synthesized facilitates the identification of treatments

that could delay or prevent toxin occurrence. DON data can be used to evaluate the effectiveness of

chemical treatments to control the disease or reduce DON concentration (Boyacioglu et al. 1992; Jones,

2000), to assist discovering factors that involve in toxin biosynthesis, toxin resistance and pathogenesis of

Fusarium graminearum (Seong et al., 2009), and to characterize Fusarium isolates that differ in their ability

to produce mycotoxins (Gale et al., 2005). DON analysis is the only means to prevent unacceptable

amounts of the toxin from entering human food and animal feed.

Several methods have been developed to analyze DON in small grain. Thin layer chromatography (TLC)

has been used for DON analysis (Trucksess et al., 1984; Fernandez et al., 1994). Chang et al. (1984)

described a method for rapid determination of DON using liquid chromatography (LC). Enzyme-linked

immunosorbent assay (ELISA) provides cost effective method for DON analysis (Casale et al. 1988). Gas

chromatography with electron capture detection (GC/EC) (Scott et al., 1981; Croteau et al., 1994; Tacke &

Casper 1996) and mass spectral detection (GC-MS) (Scott et al., 1981; Mirocha et al., 1998) methods have

also been developed for DON analysis in wheat and other grains. Recently, FT-Raman and Near-Infrared

spectroscopic technologies have been explored for the feasibility of rapid and non-destructive screening of

DON in grains (Liu et al., 2009; Peiris, et al, 2010). It has been proven that GC-MS provides a quick and

the most reliable measurement of DON and related mycotoxins in grains.

Research Methods:

The method we have used for analyzing DON and related mycotoxins in wheat and barley was developed in

our laboratory (Mirocha et al. 1998), and has been improved to give faster and more cost effective

measurements (Fuentes et al., 2006; Jiang et al., 2006). For mature grain, 4 g of ground sample was

extracted with 16 mL of acetonitrile/water (84/16, v/v). The extract was purified by passing through a

column packed with C18 and aluminum oxide (1/3, w/w), and then evaporated to dryness under nitrogen at

room temperature. DON in the dried extract was derivatized by TMS reagents (TMSI/TMCS = 100/1, v/v)

for GC-MS analysis using Selected Ion Monitoring (SIM) mode. Concentrations of DON and related

mycotoxins in barley or wheat samples were calculated using standard calibration curves generated with

each set of samples. Mirex (1,1a,2,2,3,3a,4,5,5,5a,5b,6-dodecachlorooctahydro-1,3,4-metheno-2H-

cyclobuta-[cd]pentalene) in each sample is used to monitor the stability of instruments as well as an

internal standard. A naturally Fusarium infected wheat or barley sample with a certain concentration of



DON was analyzed together with every 29 grain samples as an intra-laboratory quality control to check the

accuracy and repeatability of the DON measurements. Quality assurance was done by collaborative studies

with other Mycotoxin Diagnostic Laboratories on a monthly basis. High detection selectivity and sensitivity

of the mass spectrometer combining with fast GC program enable us to analyze a sample within 13 minutes

with minimum sample matrix interference. Up to one hundred samples could be analyzed in one day by one

instrument when needed.

Taking the advantage of high sensitivity and selectivity of the mass spectrometer in SIM mode, we are able

to detect DON in small size samples such as individual kernel, spikelet, head, small stem, leaf fragment,

and fungal culture extract at levels as low as 5 ng/sample, and use as little as 2 mg of a sample. This unique

service provides researchers an opportunity to investigate mycotoxin development in the early stages of

infection, study mechanisms of resistance to FHB, and characterize isolates of Fusarium graminearum that

differ in their ability to produce mycotoxins. Normally, 20 ~ 30% samples submitted to our laboratory are

in this category.

The DON analysis service will be available year around for FHB research projects. Turnaround time will be

between one week and two months depending on volumes of samples. Individual projects will not be

charged for toxin analysis.



Estimates for 2018/2019 and 2019/2020 PI Estimates Institution 2018/2019 2019/2020

Alyssa Collins 216 216 Pennsylvania State University Andrew Green 1000 1000 North Dakota State University Anne McKendry 1000 1000 university of Missouri Brian Steffenson 1800 1800 University of Minnesota Carl Bradley 800 800 University of Kentucky Clay Sneller 150 150 Ohio State University Corby Kistler 3500 3500 University of Minnesota Christina Cowger 300 300 USDA-ARS, Raleigh, NC Damon Smith 250 250 University of Wisconsin-Madison David Van Sanford 3000 3000 University of Kentucky Elias Elias 1500 1500 North Dakota State University Eric DeWolf 350 350 Kansas State University Eric Olson 1500 1500 Michigan State University Eric Stockinger 500 1000 Ohio State University Floyd Dowell 100 100 USDA-ARS, KS Frances Trail 150 150 Michigan State University Frederic Kolb 1800 1800 University of Illinois at Urbana Champaign Gary Bergstrom 750 750 Cornell University Gary Muehlbauer 300 300 University of Minnesota Guihua Bai 1000 1000 USDA-ARS, KS

Heather Kelly 100 100 University of Tennessee

Jason Wight 350 350 University of Maryland Jianli Chen 600 600 University of Idaho Jim Anderson 1000 1000 University of Minnesota Jinrong Xu 100 100 Purdue University Jochum Wiersma 100 100 University of Minnesota Juliet Marshall 435 435 University of Idaho Jyoti Shah 100 100 University of North Texas Kevin Smith 1000 1000 University of Minnesota Madeleine Smith 200 200 University of Minnesota Mark Sorrells 400 400 Cornell University Martin Chilvers 600 600 Michigan State University Martin Nagelkirk 220 220 Michigan State University

Mohamed Mergoum 700 1000 University of Georgia Nathan Kleczewski 500 500 University of Delaware Paul Murphy 1000 1000 North Carolina State University Pierce Paul 2700 2700 Ohio State University Richard Esten Mason 1500 1500 University of Arkansas Ruth Dill-Macky 2000 2000 University of Minnesota Shahryar Kianian 514 514 USDA-ARS, MN Stephen Harrison 1200 1200 Louisiana State University Yang Yen 200 200 South Dakota State University

Total 35485 36285



References: Bernardo, A.; Bai, G.H.; Yu, J.; Kolb, F.; Bockus, W.; Dong, Y. Registration of near-isogenic winter wheat germplasm contrasting in Fhb1 for Fusarium heat blight resistant. Journal of Plant Registrations, 2014, 8(1), 106-108.

Boddu, J.; Cho, S.; Kruger, W. M.; Muehlbauer, G. J. Transcriptome analysis of the barley-Fusarium graminearum. Interaction. Mol. Plant-Microbe Interact. 2006, 19, 407-417. Boyacioglu, D.; Hettiarachchy, N. S.; Stack, R. W. Effect of three systemic fungicides on deoxynivalenol (vomitoxin) production by Fusarium graminearum in wheat. Canadian J. Plant Sci. 1992, 72, 93-101. Casale, W. L.; Pestka, J. J.; Hart, L. P. Enzyme-linked immunosorbent assay employing monoclonal antibody specific for deoxynivalenol (vomitoxin) and several analogs. J. Agri. Food Chem. 1988, 36, 663-668. Chang, H. L; DeVries, J. W.; Larson, P. A.; Patel, H. H. Rapid determination of deoxynivalenol (vomitoxin) by liquid chromatography using modified Romer column cleanup. J. Assoc. Off. Anal. Chem. 1984, 67, 52-54.

Croteau, S. M.; Prelusky, D.B.; Trenholm, H. L. Analysis of trichothecene mycotoxins by gas chromatography with electron capture detection. J. Assoc. Off. Anal. Chem. 1994, 42, 928-933. Clark, A.J.; Costa, J.M.; Griffey, C.A.; Brown-Guedira, G.L.; Dong, Y.; Souza, E.J.; Murphy, J.P; Van Sanford, D.A. Registration of Scab-Resistant KY06C-11-3-10 Soft Red Winter Wheat Germplasm. Journal of Plant Registrations, 2014, 8(2), 211-216. Costa, J.M.; Bockelman, H.E.; Brown-Guedira, G.; Cambron, S.E.; Chen, X.; Cooper, A.; Cowger, C.; Dong, Y.; Grybauskas, A.; Jin, Y.; Kolmer, J.; Murphy, J.P; Sneller, C.; Souza, E. Registration of the Soft Red Winter Wheat Germplasm MD01W233-06-1 Resistant to Fusarium Head Blight. Journal of Plant Registrations, 2010, 4(3), 255-260. D’Mello, J. P. F.; Placinta, C. M.; Macdonald, A. M. C. Fusarium mycotoxins: a review of global implications for animal health, welfare and productivity. Anim. Feed Sci. Technol. 1999, 80, 183-205. Evans, C. K.; Xie, W.; Dill-Macky, R.; Mirocha, C. J. Biosynthesis of deoxynivalenol in spikelets of barley inoculated with macroconidia of Fusarium graminearum. Plant Dis. 2000, 84(6), 654-660. Fernandez, C; Stack, M. E.; Musser, S. M. Determination of deoxynivalenol in 1991 U. S. winter and spring wheat by high-performance thin-layer chromatography. J. Assoc. Off. Anal. Chem. 1994, 77, 628-630. Fuentes, R. G.; Mickelson, H. R.; Busch, R. H.; Dill-Macky, R.; Evans, C. K.; Thompson, W. G.; Wiersma, J. V.; Xie, W.; Dong, Y.; Anderson, J. A. Resource allocation and cultivar stability in breeding for Fusarium head blight resistance in spring wheat. Crop Sci. 2005, 45, 1965-1972. Gale, L. R.; O’Leary, L. E.; Bryant, J. D.; Ochocki, G. E.; Ward, T. J.; Kistler, H. C. Displacement of the native population of Fusarium graminearum in North Dakota and parts of Minnesota by a genetically divergent and more toxigenic population. Proceedings of the 2005 National Fusarium Head Blight Forum 2005, 158.



Jiang, G-L.; Dong, Y.; Lewis, Janet M.; Siler, Lee; Ward, R. W. Characterization of Resistance to Fusarium graminearum in a Recombinant Inbred Line Population of Wheat: Resistance to Fungal Spred, Mycotoxin Accumulation, and Grain Yield Loss, and Trait Relationships. Crop Sci., 2006, 46, 2590-2597. Jiang, G-L.; Dong, Y.; Shi, J.; Ward, R.W. QTL analysis of resistance to Fusarium head blight in the novel wheat germplasm CJ 9306. II. Resistance to deoxynivalenol accumulation and grain yield loss” Teror. Appl. Genet., 2007, 115, 1043-1052.

Jones, R. K. Assessments of Fusarium Head Blight of Wheat and Barley in Response to Fungicide Treatment. Plant Dis. 2000, 84(9), 1021-1030. Lemmens, M.; Scholz, U.; Berthiller, F.; Dall’Asta, C.; Koutnik, A.; Schuhmacher, R.; Adam, G.; Buerstmayr, H.; Mesterhazy, A.; Krska, R.; Ruckenbauer, P. The Ability to Detoxify the Mycotoxin Deoxynivalenol Colocalizes With a Major Quantitative Trait Locus for Fusarium Head Blight Resistance in Wheat. Molecular Plant-Microbe Interactions, 2005, 18(12), 1318-1324.

Leonard, K. J.; Bushnell, W. R., Eds. Fusarium Head Blight of Wheat and Barley; APS Press: St. Paul, MN, 2003. Lewis, J.M.; Siler, L.; Souza, E.; Ng, P.K.W; Dong, Y.; Brown-Guedira, G.; Jiang, G.L.; Ward, R.W. Registration of ‘Ambassador’ Wheat. Journal of Plant Registrations, 2010, 4(3), 195-204. Lewis, J.M.; Siler, L.; Souza, E.; Ng, P.K.W; Dong, Y.; Jiang, G.L.; Ward, R.W. “Registration of ‘Coral’ Wheat. Journal of Plant Registrations, 2010, 4(3), 205-214. Lewis, J.M.; Siler, L.; Souza, E.; Ng, P.K.W; Dong, Y.; Brown-Guedira, G.; Jiang, G.L.; Ward, R.W. Registration of ‘Red Amber’ Wheat. Journal of Plant Registrations, 2010, 4(3), 215-223. Lewis, J.M.; Siler, L.; Ellis, D.; Souza, E.; Ng, P.K.W; Dong, Y.; Jiang, G.L.; Ward, R.W. Registration of ‘Jupiter’ Wheat. Journal of Plant Registrations, 2012, 6(3), 315-323. Lewis, J.M.; Siler, L.; Ellis, D.; Souza, E.; Ng, P.K.W; Dong, Y.; Brown-Guedira, G.; Jiang, G.L.; Ward, R.W. Registration of ‘MSU E5024’ Wheat. Journal of Plant Registrations, 2012, 6(3), 333-341. Lewis, J.M.; Siler, L.; Ellis, D.; Souza, E.; Ng, P.K.W; Dong, Y.; Brown-Guedira, G.; Marshall, D.; Kolmer, J.; Jiang, G.L.; Ward, R.W. Registration of ‘Red Ruby’ Wheat. Journal of Plant Registrations, 2012, 6 (3), 324-332. Liu, Y.; Delwiche, S. R.; Dong, Y. Feasibility of FT-Raman spectroscopy for rapid screening for DON toxin in ground wheat and barley. Food Additives and Contaminants, 2009, 1-6.

Miller, J. D.; Arnison, P. G. Degradation of deoxynivalenol and Fusarium head blight resistance in spring cereals. J. Phtopathol. 1986, 8,147-150. Mirocha, C. J.; Kolaczkowski, E.; Xie, W.; Yu, H.; Helen, H. Analysis of deoxynivalenol and its derivatives (batch and single kernel) using gas chromatography/mass spectrometry. J. Agric. Food Chem. 1998, 46, 1414-1418. Pestka, J. J.; Smolinski, A. T. Dexoxynivalenol: toxicology and potential effects on humans. J. Toxicol. Environ. Health 2005, Part B, 8, 39-69.



Pestka, J.J. Deoxynivalenol: Toxicity, mechanisms and animal health risks. Anim. Feed Sci. Technol. 2007, 137, 283-298. Pestka, J.J. Deoxynivalenol: mechanisms of action, human exposure, and toxicological relevance. 2010, 84, 663-679.

Peiris, K.H.S; Pumphrey, M.O.; Dong, Y.; Maghirang, E.B.; Berzonsky, W.; Dowell, F.E. Near-Infrared Spectroscopic Method for Identification of Fusarium Head Blight Damage and Prediction of Deoxynivalenol in Single Wheat Kernels. Cereal Chemistry Cereal Chemistry, 2010, 87(6), 511-517. Scott, P. M.; Lau, P. Y.; Kanhere, S. R. Gas chromatography with electron capture and mass spectrometric detection of deoxynivalenol in wheat and other grains. J. Assoc. Off. Anal. Chem. 1981, 64, 1364-1371. Seong, K-Y; Pasquali, M.; Zhou, X.; Song, J.; Hilburn, K.; McCormick, S.; Dong, Y.; Xu, J-R; Kistler, H.C. Global gene regulation by Fusarium transcription factors Tri6 and Tri10 reveals adaptations for toxin biosynthesis. Mol. Microbio. 2009, 72, 354-367. Smith, K. P.; Evans, C. K.; Dill-Macky, R.; Gustus, C.; Xie, W.; Dong, Y. Host genetic effect on deoxynivalenol accumulation in Fusarium head blight of barley. Phytopathology 2004, 94, 766-771. Tacke, B. M.; Casper, H. H. Determination of deoxynivalenol in wheat, barley and malt by column cleanup and gas chromatography with electron capture detection. J. Assoc. Off. Anal. Chem. Int. 1996, 472-475. Trucksess, M. W.; Nesheim, S.; Eppley, R. M. Thin layer chromatographic determination of deoxynivalenol in wheat and corn. J. Assoc. Off. Anal. Chem. 1984, 67, 40-43. Van Sanford, D.A.; Clark, A.J.; Hershman, D.; Brown-Guedira, G.L.; Cowger, C.; Dong, Y.; Baik, B.K. “Registration of ‘Pembroke 2014’ soft red winter wheat”, Journal of Plant Registrations, 2016, 10 (1), 41-46 (DOI: 10.3198/jpr2015.07.0045crc). Wang, R.; Anderson, J.A.; Zhang, J.; Zhao, W.; Wheeler, J.; Klassen, N.; See, D.R.; Dong, Y. “Genome-Wide Association Mapping of Fusarium Head Blight Resistance in Spring Wheat Lines Developed in Pacific Northwest and CIMMYT”, Phytopathology,2017 ( DOI: 10.1094/PHYTO-02-17-0073-R). Wang, Y. Z.; Miller, J. D. Effects of Fusarium graminearum metabolites on wheat tissue in relation to Fusarium head blight resistance. J. Phytopathol. 1988, 122, 118-125. Yu, J.-B.; Bai, G.-H; Zhou, W.-C.; Dong, Y.; Kolb, F. L. Quantitative Trait Loci for Fusarium Head Blight Resistance in a Recombinant Inbred Population of Wangshuibai/Wheaton. Phytopathology, 2008, 98, 87-94. Facilities and Equipment:

The major instruments used for DON analysis will be Shimadzu QP2010 and QP2010 PLUS GC/MS systems

equipped with auto-samplers (capacity of 150 samples). Other instruments include four laboratory mills for

sample grinding, four shakers for mycotoxin extraction, three balances for sample weighing, one analytical

balance for small samples and toxin standard preparation, and three freezers to store toxin standards, reagents



and sample extracts. Mycotoxin Laboratory has enough space for hosting all the instruments and for

processing grain samples.



BUDGET JUSTIFICATION


Principal Investigator: Yanhong Dong

Total USWBSI Recommended Amount for FY18: $ 274,009

ARS Award Amount for FY18: $ 265,513

Instructions: Complete all applicable sections below where funds are being requested; description (left columns) and requested amount (right column). If budget category is not applicable, leave line item blank. NOTE: All amounts must be rounded to the nearest whole number. A. SENIOR/KEY PERSON: In fields below, add details for salary and fringe benefits associated withthe Senior/Key Person (i.e. PI/PD). Details should include PI’s Base Salary ($), the number ofCalendar, Academic and/or Summer months/time to be devoted to the funding. Provide subtotals forboth ‘Salary’ and ‘Fringe Benefits’ to the right of the descriptive details. The total amount requested for theSenior/Key Person category should be included in the far right column.

TOTAL $ AMT.

REQUESTED FOR

SENIOR/KEY PERSON

Salary: PI’s base salary is $81,750 and devote 80% time to the project, and ask for 35% of salary support.

$28,613 $38,398

Fringe Benefits: 34.2% $9,785

B. OTHER PERSONNEL: For each sub category listed below, add details for salary and fringebenefits associated with that sub category. Details should include the percentage of time (months)/totalhours to be devoted to the funding, rate of pay and fringe rate. Include the amounts requested for Salary,Fringe Benefits and number of personnel for each subcategory (Post Doc, Graduate Students,Undergraduate Students, etc.) as well as the total amount. The TOTAL amount requested for ALL ‘OtherPersonnel’ should be entered in the far right column.

TOTAL $ AMT.

REQUESTED FOR OTHER PERSONNEL

Sub Total $ Amts. Request for Salary and Fringe Benefits

Total $Amt. Requested per

Sub Category(ies)

$117,440

Post Doctoral Associates $ Salary: $ Fringe Benefits: $

Number of Post Doc Personnel: Graduate Students. NOTE: Graduate Student Tuition/Fees/Health Insurance should be included in section ‘Participant/Trainee Support Costs’ (E1).

$

Salary: $ Fringe Benefits: $

Number of Graduate Student Personnel: Undergraduate Students $63,960

Salary: 6 undergrads, 50%, $10.25/hr $63,960 Fringe Benefits: $

Number of Undergraduate Student Personnel: 6 Secretarial/Clerical $

Salary: $ Fringe Benefits: $

Number of Secretarial/Clerical Personnel: Other – Research Technician $53,480

Salary: One technician, 12 months, $41,651 $41,651 Fringe Benefits: 28.4% $11,829

Number of Other – Research Technician Personnel: 1



B. OTHER PERSONNEL (cont.) Sub Total $

Amts. Request for Salary and Fringe Benefits

Total $Amt. Requested per

Sub Category(ies)

Other – Temporary Labor $ Salary: $ Fringe Benefits: $

Number of Other – Temporary Labor Personnel:

Other $ Salary: $ Fringe Benefits: $

Number of Other Personnel:

C. EQUIPMENT: List below any items whose total dollar amount exceeds $5,000 and has a useful life of one year or more. Justification must include relevance to proposed research and dollar amounts. Include cost per item if more than one item will be purchased AND the total amount requested for this budget category in right column.

TOTAL $ REQUESTED

FOR EQUIPMENT

$

D. TRAVEL: Travel costs are the projected expenses for transportation, lodging, subsistence, and related items incurred by employees who are in travel status on official business related to the Federal award. This category is only for cooperator staff travel. Provide requested amount for domestic and foreign travel (middle $ column) in addition to the ‘Total $ Requested for Travel’ (left $ column). The travel costs should be supported with the purpose of the travel, the estimated amount of the trip(s) and the destination(s) if known at the time of award. It is not necessary to identify traveler names and travel dates.

TOTAL $ REQUESTED FOR TRAVEL

D.1. Domestic Travel (DT): List below proposed trips individually and describe their purpose in relation to the funding. Also provide dates, destination, and number of travelers where known. Include total amount per sub category below next to ‘$’and total amount requested for DT in middle column. Enter the total for Travel (DT and FT) in far right column.

Total $ Requested

for Domestic

$1,000

Research Related (e.g. travel to research plots):

$ $1,000

Non-Research Related (i.e. professional meetings): FHB Forum: For PI to attend the 2018 National FHB Forum

$1,000

Other Conferences/Meetings:

$

D.2. Foreign Travel (FT): List below proposed trips individually and describe their purpose in relation to the funding. Also provide dates, destination, and number of travelers where known. Include total amount per sub category below and total amount requested for FT in column on the right.

Total $ Requested for Foreign

Research Related (e.g. travel to research plots): $ $

Non-Research Related (i.e. professional meetings): $



E. PARTICIPANT/TRAINEE SUPPORT COSTS (P/TSC): Participant support costs means direct costs for items such as stipends or subsistence allowances, travel allowances, and registration fees paid to or on behalf of participants or trainees (but not employees) in connection with conferences, or training projects. The cost of training and education provided for employee (i.e. Graduate and Undergraduate Students) development is allowable. Tuition remission is NOT an allowable cost. Include total amount per sub category below next to ‘$’ and total amount requested for ‘Participant/Trainee Support Costs’ in column on the right (i.e. Total $ Requested).

TOTAL $ REQUESTED FOR P/TSC

1. Fees/Health Insurance:

$ $

2. Stipends:

$

3. Travel:

$

4. Subsistence:

$

5. Other:

$

F. OTHER DIRECT COSTS (ODC): This section contain multiple sub categories. Totals per sub category are required in addition to the total requested for Other Direct Costs (far right column). If there are additional sub categories under the main sub categories (i.e. Materials and Supplies), provide a total as well. F.1. Materials and Supplies (M/S): In the space below, provide as much detail and specificity as possible for all materials and supplies associated with proposed research. Materials and Supplies should be described in detail e.g., chemical reagents, printer/field paper and supplies, glassware, lumber, etc. under each sub category (Field, Greenhouse, Laboratory and Other). Include total amount per sub category below next to ‘$’ and total amount requested for M/S in the middle column (i.e. Total $ Amt. Requested – M/S)

Total $ Amt. Requested - M/S

TOTAL $ AMT. REQUESTED -

ODC Field:

$ $95,675 $108,675

Greenhouse:

$

Laboratory: Acetonitrile, isooctane, water, ethanol, acetone, TMSI, TMCS, mirex, C18, Al2O3, N2, He, plastic syringes, filter paper, pipette tips, weigh boat & paper, centrifuge tubes, test tubes, transfer pipettes, glass vials, GC vials, 200 ul insert, septa for GC vials, GC columns, GC septa, injection liners, pump filters, filaments, ion source assembly & pre-rods, masks, gloves, paper towels, lab wipes, notebooks, etc.

$95,675

Other:

$

F.2. Publications and Printing Costs (PPC): Below, provide details for any publication costs for electronic and print media, including distribution, promotion, and general handling, for which funds are being requested. NOTE: Page charges for professional journal publications are allowable provided publications report research that was supported by USDA-ARS.

Total $ Amt. Requested - PPC

$

F.3. Consulting Services (CS): For each consultant, list below the services he/she will perform, total number of days, travel costs, and the total estimated costs. Please include names and organizational affiliations for all consultants, other than those involved in consortium/contractual arrangements.

Total $ Amt. Requested - CS

$



F. OTHER DIRECT COSTS (ODC) (cont.)

F.4. Automatic Data Processing /Computer Services (ADP/CS): This section covers cost of computer services, including computer-based retrieval of scientific, technical, and education information. In the space below, list all ADP/CS and include the established computer service rates, if applicable.

Total $ Amt. Requested -

ADP/CS

$

F.5. Subawards/Consortium/Contractual Costs (SCCC): In the space below, provide details for all costs associated with subawards, consortium and contractual costs. The total requested amount for this sub-category should include both direct and indirect costs for all subaward/consortium organizations. A separate budget for the subaward should be included (i.e. attached to funding application).


SCCC $

F.6. Equipment/Facility/Land Rental and User Fees (RUF): List the total funds requested for equipment or facility rental/user fees. Justify each rental user fee by providing specific details (e.g. Land Rental Fees – number of acres/cost per acre).

Total $ Amt. Requested –

RUF $

F.7. Alterations and Renovations (A&R): List the total funds requested for alterations and renovations (A&R). Justify (i.e. required in order to carry out research) the costs of alterations and renovations, including repairs, painting, and removal or installation of partitions, shielding, or air conditioning. Where applicable, provide the square footage and costs.

Total $ Amt. Requested –

AR

$

F.8. Other - Miscellaneous Direct Costs (OMDC): Under each relevant sub category below, enter a brief description, and basis for the estimate (i.e. individual fee rate/price). Include total amount per sub category below next to ‘$’ and total amount requested for ODC in column on the right.


OMDC Laboratory Animal Fees: $ $13,000 U.S. based Winter Nurseries: $

International Nurseries: $ Double Haploids: $ Other Analyses/Services: $

Communication (postage, shipping, fax, long distance phone): $ Photocopying: $

Other MODC (describe): $13,000 for maintaining 2 GC-MS instruments ($10,000), 4 sample mills ($1,000), 4 shakers ($1,000), 4 balances ($500) and pipettes ($500).

$13,000



H. INDIRECT COSTS (IDC): Provide below your Institution’s approved Indirect Cost (IDC) rate for

USWBSI/USDA-ARS grants. ‘Type’ refers to Total Costs (TC), Direct Costs (DC), Modified Direct Costs (MDC). Rate and Type has been prefilled by NFO based on submitted FY18 Pre-Proposals.

TOTAL $ AMT.

REQUESTED FOR IDC

IDC Rate and Type: N/A for NACAs IDC Base Amount:

$ N/A

J. FEE (Small Business Act – SBIR Fee): The SBIR fee is a Congressional mandated fee charged to all ARS/USWBSI grants and is applicable to all non-ARS PIs. The rate for FY18 is 3.2% and will be deducted from the USWBSI’s recommended amount prior to the processing of the award by ARS. The Formula for calculating the fee is below (prefilled by the NFO):

Step 1 – USWBSI’s Total Recommended Amount/1.032 Step 2 - Result from Step 1 should be subtracted from USWBSI Recommended Amount to obtain the SBIR

Fee).

TOTAL $ AMT FOR FEE-SBIR

Step 1: 274,009 / 1.032 = 265,513 Step 2: 274,009 - 265,513 = 8,496

$ 8,496



PROJECT BUDGET PAGE

Instructions: Insert values from corresponding budget justification into this form.

Budget based on OMB Number: 4040-0001

PROJECT 1 TITLE: Diagnostic Services for DON. PROJECT ID: FY18-DO-002

ARS AGREEMENT NO: New

Totals ($)

PRINCIPAL INVESTIGATOR: Yanhong Dong

ORGANIZATION: University of Minnesota A. Senior/Key Persons (i.e. PI/PD) ........................................................................................................................................ ® $38,398 B. Other Personnel (Post-Docs, Graduate Students, Secretarial/Clerical, Research Technician, Temporary Labor, and

Other) ................................................................................................................................................................................................... ® $117,440

Total Number of ‘Other Personnel’: 7

Total Salaries, Wages and Fringe Benefits (A + B) ................................................................................................ ® $155,838

C. Equipment ................................................................................................................................................................................. ®

D. Travel (Insert total amount for D. Travel to left and totals for subsections (1 and 2) below) ............................... ® $1,000 1. Domestic .................................................................................................................................................. ® $1,000

2. Foreign ...................................................................................................................................................... ®

E. Participant/Trainee Support Costs (Insert total for E to left and totals for sub sections below) ........... ® 1. Fees/Health Insurance .......................................................................................................................... ® $ 2. Stipends .....................................................................................................................................................® 3. Travel .........................................................................................................................................................® 4. Subsistence ...............................................................................................................................................® 5. Other...........................................................................................................................................................®

Total Number of Participants/Trainees: F. Other Direct Costs (Insert total amount for F) ............................................................................................................. ® $108,675

1. Materials and Supplies ......................................................................................................................... ® $95,675 2. Publication Costs ................................................................................................................................... ® 3. Consultant Services ............................................................................................................................... ® 4. ADP/Computer Services ...................................................................................................................... ® 5. Subawards/Consortium/Contractual Costs ..................................................................................... ® 6. Equipment or Facility Rental/User Fees .......................................................................................... ® 7. Alterations and Renovations ............................................................................................................... ® 8. Other - Miscellaneous........................................................................................................................... ® $13,000

G. Total Direct Costs (Total Salaries, Wages and Fringe thru F).................................................................................® $265,513

H. Indirect Costs not applicable for NACAs ....................................................................................................................®Rate and Type: N/A Base:

N/A

I. Total Direct and Indirect Costs - ARS FY18 Award Amount (G + H) ............................................ ® $ 265,513 J. FEE - Small Business Act – SBIR Fee (3.2%).................................................................................................... ® $ 8,496

K. TOTAL COSTS - USWBSI FY18 Total Recommended Amount (I + J) .................................... ® $ 274,009

NAME AND TITLE SIGNATURE

(Adobe E-Sign or insert image of signature) DATE Principal Investigator

Yanhong Dong 07/03/2018



ADJUSTMENT SUMMARY PAGE

USWBSI Consolidate Award Title: Diagnostic Services for DON

Principal Investigator: Yanhong Dong Institution: University of Minnesota

Fiscal Year: 2018 USWBSI’s FY18 Total Recommended Amount: $ 274,009

ARS Agreement Number: New USDA-ARS FY18 Total Award Amount: $ 265,513

Instructions: Under each of the project titles listed below, please indicate the changes made to the pre-proposal that address the comments given by the Review Panel(s) and/or the Executive Committee (see table in “Letter of Instructions”). Use additional pages if necessary.


None.


I agree to send copies of any printed materials (e.g. brochures, extension publications, etc.) and/or electronic versions of communication materials or URL links to materials posted on the Web to the Networking & Facilitation Office of the U.S. Wheat & Barley Scab Initiative.

______________________________________ Principal Investigator Date


COMMUNICATION PLAN

USWBSI Consolidate Award Title: Diagnostic Services for DON

Principal Investigator: Yanhong Dong Institution: University of Minnesota

Fiscal Year: 2018 USWBSI’s FY18 Total Recommended Amount: $ 274,009

USDA-ARS Agreement Number: New USDA-ARS FY18 Total Award Amount: $ 265,513

Instructions: Using the space below, describe in detail how you plan to communicate the results from this research to your stake-holders in the most effective way. Please describe your target audience (i.e. USWBSI Administration/members, industry, private growers, interest groups etc.) and the methods (i.e. written, electronic, oral, etc.) of communication you will use to communicate your results to your audience.

The target audiences of this project are the Fusarium head blight (FHB or scab) researchers. We will provide DON results to the researchers in MS Excel file via email and answer their questions through either email or phone.

07/03/2018

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