USDA-ARS/ U.S. Wheat and Barley Scab Initiative FY07 Final Performance Report (approx. May 07 April 08) July 15, Cover Page

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1 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Tilahun Abebe Institution: University of Northern Iowa Address: Department of Biology 144 McCollum Science Hall Cedar Falls, IA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 32,664 Engineering Barley with Antifungal Gene Gastrodianin to Enhance Resistance to Scab Disease. USWBSI Individual Project(s) USWBSI Research Area * GET ARS Adjusted Award Project Title Amount Engineering Barley with Anti-Fungal Gene Gastrodianin for $32,664 Resistance to Scab Disease. Total Award Amount $ 32,664 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

2 PI: Abebe, Tilahun USDA-ARS Agreement #: Project 1: Engineering Barley with Anti-Fungal Gene Gastrodianin for Resistance to Scab Disease. 1. What major problem or issue is being resolved and how are you resolving it? There is very little genetic resistance to Fusarium head blight (FHB) in barley. Genetic engineering has a great potential in the development of FHB resistant genotypes. So far, however, engineering resistant barley has been unsuccessful because of lack of genes that effectively inhibit Fusarium graminearum. Transformation with the common pathogenesisrelated proteins such as, chitinases, glucanases, and thaumatin-like proteins (TLPs), did not significantly improve resistance. There is a need to study efficacy of other fungal resistance genes against F. graminearum. We are working with an antifungal gene gastrodianin isolated from a Chinese medicinal plant Gastrodia elata. Gastrodianin is known to inhibit growth of F. graminearum and other saprophytic fungi. The major routes of F. graminearum infection of barley kernels are through the husk (lemma and palea) and the apex of florets. To successfully block infection, expression of gastrodian in transgenic Golden Promise barley was targeted to the husk and epidermis of the kernel using a spike-specific Lem2 promoter that was isolated from Morex barley. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Golden Promise barley was co-transformed with plasmids plem2vgm2 (containing GFP and gastrodianin) and pahc25 (containing a selectable bar gene). Both GFP and gastrodianin are driven by the spike-specific Lem2 promoter. Fifteen T 0 transformants were recovered. Of these, eleven were fertile, two plants were sterile and two plants were lost to contamination. Integration of gastrodianin into the genome of Golden Promise was verified by PCR. Accumulation of gastrodianin transcript was confirmed by RT-PCR. Western analysis using a polyclonal antibody raised against gastrodianin protein also detected a band at 12 kda, which corresponds to the size of the mature protein. We planned to test resistance of transgenic plants to F. graminearum in fall 2007 and spring of Unfortunately, the transgenic plants grow very slow and we were unable to collect seeds for planting T1 and T2 generation. Instead, greenhouse testing is scheduled for fall Impact: Use of the tissue-specific Lem2 prompter will direct accumulation of gastrodianin protein in the spike of transgenic plants. This is advantageous because the protein will only be expressed in the tissues F. graminearum must cross to infect the kernel. In addition, constitutive expression of proteins is an energy-demanding process and can interfere with metabolic processes resulting in dwarf plants. Restricting expression of gastrodianin to the spike could reduce possible side effects of the protein on growth. (Form FPR07) 2

3 PI: Abebe, Tilahun USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Transgenic Golden Promise plants expressing gastrodianin can be crossed to elite barley lines to develop varieties that are resistant to F. graminearum. Ultimately, resistant varieties can increase yield and quality of grain in areas where scab disease is a major problem. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None (Form FPR07) 3

4 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Eric Adee Institution: University of Illinois Address: Northwestern Illinois Agricultural Research and Demonstration Center th Ave. Monmouth, IL Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Uniform Fungicide Trial for Scab on Wheat in Illinois. Title: FY07 ARS Award Amount: $ 4,878 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Uniform Fungicide Trial for Scab on Wheat in Illinois. $4,878 Total Award Amount $ 4,878 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

5 PI: Adee, Eric USDA-ARS Agreement #: Project 1: Uniform Fungicide Trial for Scab on Wheat in Illinois. 1. What major problem or issue is being resolved and how are you resolving it? Head scab on wheat has caused severe losses in yield and grain quality in Illinois. Fungicides have the potential to be one of the best options for control of scab (Fusarium head blight, FHB), but many fungicides are not affective or are not labeled for application time necessary for control of scab. The goal of this study is to test the efficacy of new or recently labeled fungicides and biological control agents in controlling head scab on wheat and their resulting effect on yield and quality of wheat. A small reduction in the toxin level (DON) produced by the fungus means a great deal to the millers who process the grain. Conducting uniform fungicide trials will give farmers valuable information for management of scab as well as contributing to data applicable across all the Midwestern wheat growing regions. Wheat for these trials were planted at Brownstown, DeKalb, Dixon Springs and Monmouth, IL. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Weather conditions were not favorable for FHB development to adequately test the effectiveness of the products. The study at Brownstown had no visible symptoms of FHB, however, the level of DON reduced significantly by all but one fungicide (Table 1). The percent incidence of FHB at Monmouth was very low, 4.75, in the check with no fungicides, indicating there was very little disease pressure from FHB. There were no differences between the check and the fungicide treatments for FHB severity, incidence, grain yield, or level of DON at Monmouth (Table 2). There were no leaf disease ratings taken at Monmouth due to the dry conditions experienced after heading resulting in very healthy leaves. These dry conditions may have affected the production of DON, resulting in very low levels the control plot and with all fungicide treatments. The studies at DeKalb and Dixon Springs were lost due stand quality issues associated with winter-kill. These results have been presented through Extension activities and can aid companies in developing products and farmers in selecting fungicides that will be effective in controlling scab. (Form FPR07) 2

6 PI: Adee, Eric USDA-ARS Agreement #: Table 1: Uniform FBH Fungicide Trial, Brownstown, IL, 2007 Fungicide Treatment z Test Weight (lb/bu) Yield (bu/ac) DON v (ppm) Untreated check Folicur 432SC 4.0 fl oz Caramba 8.2 fl oz Caramba 13.5 fl oz Topguard 14 fl oz Prosao 6.5 fl oz Proline 5 fl oz Pr>F LSD (0.05) (0.49) Table 2: Uniform FBH Fungicide Trial, Monmouth, IL, 2005 Fungicide Treatment z FHB FHB Severity FHB Index w Yield Test Incidence (%) x (bu/ac) Weight DON v (%) y (lb/bu) (ppm) Untreated check Folicur 432SC 4.0 fl oz Caramba 8.2 fl oz Caramba 13.5 fl oz Topguard 14 fl oz Prosao 6.5 fl oz Proline 5 fl oz Tilt 4 fl Pr>F z Fungicides were applied at the early flowering stage (Feekes 10.51) with 0.125% Induce.. y Incidence is the percent of heads with symptoms of FHB. xseverity is percent of florets with FHB in the infected heads. w Disease index is FHB incidence x severity. v Deoxynivalenol Impact: Foliar fungicides will have little impact on yield of wheat if the environmental conditions are not favorable for the development of fungal diseases attacking the head and leaves before early grainfill. It does appear that several of these fungicides can lower the production of DON even if there are very few symptoms of FHB visible on the heads. (Form FPR07) 3

7 PI: Adee, Eric USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Data from these studies show that there was relatively little benefit in applying fungicides to wheat in northern Illinois in Utilizing disease prediction models that include weather data, variety characteristics, cropping history and growth stage can greatly benefit growers when determining if fungicide applications are economically viable. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. (Form FPR07) 4

8 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Tika Adhikari Institution: North Dakota State University Address: Department of Plant Pathology 331 Walster Hall Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 34,297 Effect of Inoculum Levels, Host Resistance, Fungicide and Weather on FHB. USWBSI Individual Project(s) USWBSI Research Area * EEDF HGG ARS Adjusted Award Project Title Amount Effects of Growth Stage, Host Resistance, and Weather on DON and FHB Development. $24,939 Characterizing the FHB Resistance of 'Frontana' Wheat Using Unique RBCM Lines. $ 9,358 Total Award Amount $ 34,297 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

9 PI: Adhikari, Tika USDA-ARS Agreement #: Project 1: Effects of Growth Stage, Host Resistance, and Weather on DON and FHB Development. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (Scab), caused by Fusarium graminearum, is one of the important diseases of wheat worldwide. Although wheat cultivars moderately resistant to FHB is managed through fungicide applications and cultural practices, a reliable disease forecasting system is essential to predict FHB epidemics during the season for the wheat growers to know if their crop needed fungicide application. 2. What were the most significant accomplishments and impact? Accomplishment: To determine effects of three wheat cultivars Glenn (FHB resistant), Steele-ND (moderately susceptible) and Trooper (susceptible) and crop growth stages on FHB severity and incidence, and DON content, field plots were artificially inoculated at Fargo, ND. Glenn has the lowest (20.6%) and Trooper has the highest FHB severity (28.12%). All three disease components: incidence (12.75%), severity (41%), and DON (2.45 ppm) were higher when the cultivars were inoculated at mid flowering stage. A positive correlation was observed between FHB severity and DON content in all three cultivars Glenn (r = ), Steele-ND (0.9893), and Trooper (0.9844). It appeared that locally available G. zeae inoculum could play a significant role in disease development, and infection at mid flowering growth stage could be a crucial in FHB incidence, severity, and DON production. Impact: This project would ultimately speed up the efforts in the development of effective and précised forecaster, and would help wheat producers better manage disease. Incorporation of FHB severity data into the disease forecasting system would help in DON level prediction prior to harvesting. 3. A list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles. Ali, S., and T. B. Adhikari Effect of wheat genotypes and inoculation timings on Fusarium Head Blight of wheat in North Dakota. Page 24: in the Proceeding of the 2007 National Fusarium Head Blight Forum, December 2-4, 2007, The Westin Crown Center, Kansas City, MO. Paul, P. A., P. E. Lipps, E. De Wolf, G. Shaner, G. Buechley, T. Adhikari, S. Ali, J. Stein, L. Osborne, and L. V. Madden A distributed lag analysis of the relationship between Gibberella zeae inoculum density on wheat spikes and weather variables. Phytopathology 97: Paul, P. P., L. V. Madden, S. Wegulo, T. Adhikari, S. Ali, and E. DeWolf Influence of SRWW, HRSW, and HRWW varieties on the relationship between FHB and DON. Page 128: in the Proceeding of the 2007 National Fusarium Head Blight Forum, December 2-4, 2007, The Westin Crown Center, Kansas City, MO. Nita, M, E. DeWolf, L. Madden, P. Paul, S. Shaner, T. Adhikari, S. Ali, J. Stein, and L. Osborne, and S. Wegulo Mechanistic simulation models for Fusarium Head Blight and DON. Page 108: in the Proceeding of the 2007 National Fusarium Head Blight Forum, December 2-4, 2007, The Westin Crown Center, Kansas City, MO. (Form FPR07) 2

10 PI: Adhikari, Tika USDA-ARS Agreement #: Project 2: Characterizing the FHB Resistance of 'Frontana' Wheat Using Unique RBCM Lines. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) of small grains, is caused primarily by Fusarium graminearum Schwabe. Both yield and quality are severely affected by the fungal mycotoxins [(e. g., deoxynivalenol (DON)]. The Brazilian spring wheat cultivar, Frontana represents a genetically different source of resistance and prevents initial fungal infection (Type I resistance) or accumulation of DON (Type V resistance). Unique reciprocal backcross monosomic (RBCM) lines have been developed from a cross between the resistant cultivar Frontana and the susceptible cultivar Chris. It would be advantageous to wheat breeding efforts if this mechanism was characterized and if the chromosomes carrying the genes involved in the Frontana type of resistance could be confirmed. 2. What were the most significant accomplishments and impact? Accomplishment: An attached-leaf bioassay and real-time PCR analysis were used to determine resistance in the two parents Frontana (resistant parent), Chris (susceptible parent), Frontana chromosome lines (FTN 3AS, FTN 6AS, and FTN 4DS), and Alsen using digital image analysis. After disease scoring, the inoculated leaf segments for each cultivar/line were used to quantify fungal biomass by real-time PCR assays. Frontana and three lines (FTN 3AS, FTN 4DS, and FTN 6AS) exhibited smaller lesions than Alsen and Chris. Real-time PCR assay detected the highest FgDNA in Chris ( pg) and the lowest FgDNA in FTN 4DS line (0 pg). FgDNA in Frontana and other two lines (FTN 3AS and FTN 6AS) ranged from 0.75 pg to 0.98 pg. Both leaf bioassay and real-time PCR analysis will provide accurate quantification of F. graminearum in wheat and can be useful to differentiate among susceptible and resistant wheat lines to early infection of FHB. Impact: The resistant line(s) identified in this study can be utilized in wheat breeding programs for the development of FHB-resistant wheat cultivars. (Form FPR07) 3

11 PI: Adhikari, Tika USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None. (Form FPR07) 4

12 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: James Anderson Institution: University of Minnesota Address: Department of Agronomy and Plant Genetics 411 Borlaug Hall 1991 Upper Buford Circle St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 219,091 Breeding and Development of DNA Markers for Fusarium Head Blight Resistance in Wheat. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGR Characterization and Pre-Breeding of Novel Fusarium Head Blight Resistance Sources. $54,974 HGG Construction and Utilization of a Pooled BAC Library of Sumai 3. $ 29,775 HGG QTL Mapping of Wheat Fusarium Head Blight Resistance in the Japanese Landrace PI $ 50,732 VDUN Breeding Fusarium Head Blight Resistant Spring Wheat. $ 83,610 Total Award Amount $ 219,091 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

13 PI: Anderson, James USDA-ARS Agreement #: Project 1: Characterization and Pre-Breeding of Novel Fusarium Head Blight Resistance Sources. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight resistant spring wheat germplasm has been identified by vigorous screening of the worldwide collections in the National Small Grains Collection. The utilization of the selections is not satisfactory due to the lack of adequate information on their novelty, and the low direct breeding value of the unadapted germplasm. This project confronts the issue of characterizing the resistance, introgression of the resistance into elite germplam, and finding additional new sources of resistance in spring wheat. We use FHB nurseries to select for FHB resistance and adaptability in spring wheat germplasm introduction and pre-breeding populations. Resistant selections made based on three year replicated FHB screening nurseries were evaluated for point-inoculation in the greenhouse, and genotyped with DNA markers linked to FHB resistant QTLs. The most resistant selections are distributed to breeders. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A total of 25 Elite year 2, 21 Elite year 1, 15 advanced plant introductions, 15 selected Abura/Wheaton RIL lines, 74 Wheaton/Tokai 66 RILs; 114 PI /Wheaton RILs; 130 Wheaton/ RILs; 138 Wheaton/PI (Prodigio Italiano) RILs, and 112 Wheaton/PI (Chudoskaja) RILs were evaluated for FHB resistance in 2007 FHB nurseries at Crookston and/or St. Paul. The Crookston nursery had high disease levels and we were able to make selections within and among the most promising populations. Due to low disease levels in the St. Paul nursery, presumably due to unseasonably high temperatures, we did not get useful, discriminating data. Based on this data, we planted 318 RILs from 5 different populations, and 4 selected RILs (Wheaton/Tokai 66 RIL57, Wheaton/ RIL53, Wheaton/Abura RIL64, Wheaton/Abura RIL83) in our 2008 Crookson FHB nursery. Six F 3 populations were also planted in 2008 for generation advancement as we continue to cross the best resistance sources with adapted spring wheat lines. These six populations are: Wheaton/PI 19766, Wheaton/PI 62083, Wheaton/PI 69261, RB07/PI 19766, RB07/PI 62083, and RB07/PI Impact: We have continued our characterization of potential new FHB resources and screening of the most promising materials that were crossed to regionally adapted HRS lines. This makes the FHB resistance genes contained in these materials more accessible to the breeding community because they are contained in improved genetic backgrounds. Furthermore, we have prioritized our pre-breeding efforts to focus on materials that do not contain Fhb1 based on diagnostic markers. This allows us to focus on germplasm more likely to contain novel resistance genes. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: All germplasm and pre-breeding materials are available upon request. RIL populations developed as part of this project are good candidates for QTL mapping populations. Our Wheaton/PI population (Project 3 of this report), resulted from this project. After (Form FPR07) 2

14 PI: Anderson, James USDA-ARS Agreement #: summarizing multiple years of data for these materials, the results will be published, similar what is shown in Crop Sci. 48: which summarizes earlier work on this project. Accomplishment: Five lines, resulting from this project were entered into the 2007 Uniform Regional Scab Nursery. All five were from the cross Wheaton/Abura. Two of the RILs, namely #64 and #83 had the lowest overall DON among the 48 entries in the test. Impact: Abura is a good source of FHB resistance, but is not well adapted to the HRS wheat region of the U.S. We have incorporated high levels of FHB resistance from this source into a more adapted HRS background by crossing it with Wheaton. The high levels of resistance in progeny from this cross indicate that high levels of FHB resistance can be recovered from single crosses involving a highly susceptible parent. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The regional scab nursery provides an effective means of sharing germplasm. Therefore, other regional breeding programs growing this nursery have had these materials and opportunity to cross with them since April, (Form FPR07) 3

15 PI: Anderson, James USDA-ARS Agreement #: Project 2: Construction and Utilization of a Pooled BAC Library of Sumai What major problem or issue is being resolved and how are you resolving it? Bacterial Artificial Chromosome (BAC) libraries are an essential tool for gene cloning and sequence analysis. There are no BAC libraries of wheat available in the U.S. and none of the few existing libraries worldwide were constructed using a known FHB resistance source. The main purpose of this grant it to develop genomics resources that will be useful in our cloning of Fhb1, the major QTL for Fusarium head blight resistance in wheat. A BAC library of Sumai 3, the donor of Fhb1 and at least two other important FHB resistance QTL is being constructed. This library may be useful to other groups attempting to clone other FHB resistance from Sumai 3. The objectives of the grant are to: 1) Construct a pooled BAC library of Sumai 3 for the wheat community. 2) Construct a Sumai 3 BAC contig spanning Fhb1. 3) Develop user-friendly markers for Fhb1 and provide the markers to the Regional Genotyping Centers. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Objective 3 has been completed. Dr. Sixin Liu, postdoctoral research associate, has worked with Shiaoman Chao at the Fargo Genotyping Center in designing and testing new markers for Fhb1 that are more amenable to the equipment used at the Genotyping Centers. We have developed a robust, co-dominant marker, named UMN10 that can be used to screen for Fhb1. Impact: This marker is more diagnostic and easier to use compared to previously developed markers. This helps breeders select for the exact genotype at the Fhb1 locus. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The Fargo, Manhattan, and Raleigh Genotyping Centers and several wheat breeders in the U.S. have all been using marker UMN10 since December Wheat breeders can now more efficiently breed for resistance to FHB using this improved marker. Accomplishment: At the current stage the Sumai 3 BAC library consists of 460,800 clones with the average insert size 120 kb. This number of clones corresponds to 3.5x coverage of the hexaploid wheat genome. All BAC clones are arrayed into 384-well microtiter plates (a total of 1,200 plates) containing LB-based freezing media suitable for long-term storage of bacterial clones at -80C. The original copy of the library is currently being stored in -80C freezer in the Department of Plant Pathology, KSU. (Form FPR07) 4

16 PI: Anderson, James USDA-ARS Agreement #: Impact: None realized as yet as the BAC library is still in development. We are using FY08 funding to complete the library and begin screening clones from the library this fall and plan to sequence one or more of the BAC clones containing Fhb1 within 6 months. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: There are currently no BAC libraries of hexaploid wheat available in the US. Therefore, Sumai 3 BAC library is important addition to genomic resources available to US wheat researchers. Recent studies of wheat genome evolution demonstrated significant amount of structural variation among different wheat lines. This variation results in absence of perfect synteny between wheat accessions and precludes utilization of available BAC libraries for positional cloning of all wheat genes. Therefore, Sumai 3 BAC library will provide additional resource for positional cloning of agronomically important genes in wheat. (Form FPR07) 5

17 PI: Anderson, James USDA-ARS Agreement #: Project 3: QTL Mapping of Wheat Fusarium Head Blight Resistance in the Japanese Landrace PI What major problem or issue is being resolved and how are you resolving it? We are trying to improve FHB resistance in wheat by incorporating new resistance genes. The resistant wheat landrace, PI 81791, was identified as having high levels of FHB resistance and not containing Fhb1 as part of another USWBSI-funded project (the progenitor of FY07 Project 1). A mapping population of 150 RILs was constructed by crossing PI with the agronomically adapted line, Wheaton. Phenotypic evaluation in four different field environments and two greenhouse environments are being performed to evaluate the resistance to Fusarium graminearum. These 150 RILs will also be genotyped using SSR markers. Using both the phenotypic and genotypic data, QTL involved in resistance to scab will be identified and mapped. Identification and mapping of these QTL will allow for the resistance to be capitalized upon for use in breeding programs and, potentially, for the eventual release of lines containing this resistance to growers. Furthermore, markers will be assessed for their ability to be used diagnostically in order to identify the resistance QTL in existing and future lines. Additionally, the phenotypic screens will be used to identify RILs that are both resistant and agronomically adapted for use as parents in our breeding program. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): A.) Accomplishment: Completion of disease screenings. Type II resistance screening in the greenhouse has been completed for both the fall 2007 and spring 2008 seasons. Type I and II resistance screening in the field was completed for the summer 2007 season at both St. Paul and Crookston, MN locations. This population is being grown in 2008 FHB nurseries at both locations. Impact: The greenhouse and field resistance scores will be used as phenotypic data for QTL mapping of the Wheaton/PI mapping population. Seed from heads harvested from the field screenings are being used to assess visually scabby kernels (VSK) and determine DON content for each of the RILs and parents. Additionally, the scores are being used to identify resistant lines that are also agronomically adapted for use as parents in our breeding program. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The data collected during the 2007 and 2008 greenhouse seasons and the 2007 field season, as well as the data that will be collected upon the completion of the 2008 field season, has provided a means to more accurately identify the QTL involved in scab resistance from the PI resistance source. Additionally, the field and greenhouse screenings provide data to identify new potential parents for breeding programs that have shown consistent resistance and agronomic adaptation. (Form FPR07) 6

18 PI: Anderson, James USDA-ARS Agreement #: B.) Accomplishment: Development of QTL mapping tools. The Wheaton and PI screening with approximately 880 SSR markers was done at the USDA-ARS Fargo Genotyping Lab. The data generated from that screen resulted in the identification of 519 that are informative (i.e., polymorphic or dominant). DNA has also been isolated from all 150 individuals comprising the Wheaton/PI RIL mapping population, as well as the parents of this population. Impact: The DNA will be used for mapping the 519 polymorphic markers available for the population. A majority of these markers will be run by a Ph.D. student on the Anderson project at the USDA-ARS Fargo Genotyping Lab from mid to late August. The markers and DNA will be used to identify the QTL involved with scab resistance donated by PI via QTL mapping procedures. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The polymorphic SSR markers will allow for the identification and locating of resistance QTL. The SSR markers may also be useful for future comparative and association mapping work, as they are being consistently used on several different wheat mapping populations for various traits. Also, once QTL are identified, some of the SSR markers can potentially be used as diagnostic markers. These diagnostic markers may be useful across different populations in identifying scab resistant individuals with resistance originating from PI (Form FPR07) 7

19 PI: Anderson, James USDA-ARS Agreement #: Project 4: Breeding Fusarium Head Blight Resistant Spring Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Wheat varieties with greater resistance to Fusarium head blight (FHB) would make a substantial contribution to reducing the losses from this devastating disease. The main objective of this project is to develop Fusarium head blight resistant wheat germplasm and varieties adapted for commercial production in Minnesota and the surrounding region and characterize the level of FHB resistance of all wheat varieties grown in the region. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The new variety Tom was released in Named to honor Tom Anderson, Tom has moderate resistance to FHB and has high grain yields. The overall performance of this variety it similar to Freyr, the most popular variety among Minnesota wheat growers in However, Tom also has moderate resistance to stem rust race Ug99. This provides an additional safeguard in case this new stem rust race reaches the U.S. in the next few years. Impact: Tom is rated as a 4 for FHB resistance on our 1-9 scale in which 1 is the highly resistant and 9 highly susceptible. Currently, only six other wheat varieties in the region have a scab rating this good. Unfortunately, 4 of these 6 varieties have lower than average yields and are not likely to be popular with growers. Tom s moderate FHB resistance can help reduce losses to this disease. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The release of Tom gives wheat growers in the region an additional option when selecting for varieties that provide high yield and FHB resistance. It is vitally important that FHB resistant varieties also have high yields and acceptable agronomic characteristics and enduse quality. Tom is one of only two varieties (Freyr is the other) that combines a moderate level of FHB resistance (4 or lower on our 1-9 scale) and above-average yield among the 20 varieties for which we have at least 3 years of Minnesota yield data. Accomplishment: Five experimental lines were entered in the 2007 Uniform Regional Scab Nursery. These lines were identified in previous testing as having improved levels of FHB resistance and two of them were among the best performers in the nursery. MN , a repeat entry from the 2006 nursery had the 3 rd lowest DON level among the 48 entries in the trial and was also in the top 10 for lowest disease index and VSK. Impact: These lines combine FHB resistance from different sources and are candidates for germplasm release. These lines are available and have been requested by other wheat breeders in the region for use as crossing parents. (Form FPR07) 8

20 PI: Anderson, James USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These materials contain unique combinations of FHB resistance genes and in many cases the resistance levels are superior to the resistant checks used in the nursery. Increasingly, the other important characteristics desired of crossing parents are improving as well (e.g. shorter height, greater straw strength, better leaf rust resistance, better grain yield and quality). Accomplishment: Scab nurseries were established at 3 field sites in A total of 1,700 genotypes were evaluated in 5,000 total rows among the locations. Dry, hot weather resulted in low FHB at the Morris and St. Paul locations. The Crookston FHB screening nursery was excellent, similar to 2006, and provided highly discriminatory data. As a result of this nursery and results from previous years, the FHB resistance data of 25 spring wheat cultivars was assessed and reported to growers via print media and field day presentations. Impact: Good field screening nurseries are needed to maintain progress in breeding for FHB resistance. FHB remains a potentially devastating disease in the region as severe damage was inflicted in Our FHB resistance ratings are an important part of growers decision regarding which variety they will grow. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before: The U of MN FHB variety ratings are based on data from about six FHB inoculated, mist-irrigated nurseries (fewer than six is possible if the nurseries were highly discriminatory). In most cases, we have ratings of new varieties published by December of the year preceding the sale of certified seed to growers. (Form FPR07) 9

21 PI: Anderson, James USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-Reviewed Articles Anderson, J.A Marker-assisted selection for Fusarium head blight resistance in wheat. Intl. J. Food Microbiol. 119: Anderson, J.A, S. Chao, and S. Liu Molecular breeding using a major QTL for Fusarium head blight resistance in wheat. Crop Sci. 47:S-112-S-119. Abstracts/Proceedings Anderson, J.A Use of MAS for FHB Resistance: Is it working for Wheat Breeders? In S.M. Canty, A. Clark, D. Ellis, and D. Van Sanford (eds), Proceedings of the National Fusarium Head Blight Forum; Dec 2-4, 2007; Kansas City, MO. Okemos, MI: ASAP Printing, Inc. pp. Reports Anderson, J., J. Wiersma, J. Kolmer, and R. Dill-Macky Spring Wheat. In Preliminary Report 24; 2006 Wheat, Barley and Oat Variety Performance in Minnesota, Preliminary Report, Edited by Jochum Wiersma. Anderson, J.A., G.L. Linkert, and J.J. Wiersma Hard Red Spring Wheat. In Minnesota Varietal Trials Results, University of Minnesota Extension Service. (Form FPR07) 10

22 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Olin Anderson Institution: USDA-ARS Address: Western Regional Research Center 800 Buchanan St. Albany, CA Phone: Fax: Fiscal Year: 2007 FY07 ARS Agreement ID: NA Agreement Title: U.S. Wheat & Barley Scab Initiative's Networking and Facilitation Office and Website. FY07 ARS Award Amount: $ 35,500 USWBSI Research Area * EC/HQ USWBSI Individual Project(s) ARS Adjusted Project Title Award Amount U.S. Wheat & Barley Scab Initiative's Networking and Facilitation Office and Website. $ 35,500 Total Award Amount $ 35, Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

23 PI: Anderson, Olin ARS Agreement #: NA Project 1: U.S. Wheat & Barley Scab Initiative's Networking and Facilitation Office and Website. 1. What major problem or issue is being resolved and how are you resolving it? The Scab community requires easy to use, comprehensive, and centralized tools for data management and dissemination. These tools are provided, maintained, and created as necessary by the U.S. Wheat & Barley Scab Initiative. A suite of in-house designed and customized off the shelf software packages are hosted by a U.S. Wheat & Barley Scab Initiative server computer. Most applications are accessed through a friendly web browser based interface that can be accessed from any internet connected computer and at any time, day or night. These services include online calendars, weblogs, community mailing lists, forum registration, forum proceedings purchasing, and discussion boards. Standard web pages are also employed to provide online information sources concerning research areas, news, annual forums, literature, and useful links. Additionally, the U.S. Wheat & Barley Scab Initiative maintains a web-accessible database (the USWBSI database) that allows querying and downloading of public information. An improved feature set, better user interface, more services, and improved reliability are goals of the U.S. Wheat & Barley Scab Initiative. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment (1): Electronic Pre-Proposal Submission System Impact (1): After last year's successful partial implementation of the Electronic Pre- Proposal Submission system (EPS), it was determined that the system could be broadened to include all of the research areas for this year's Pre-Proposal submission process. User feedback from last year's beta interface was used to build a list of features and a process flow that would be easy and intuitive for first time users and to expand on the system's features. The improvements to the system are still underway but are scheduled to be finished before the Pre-Proposal Submission process starts on August 1, The beta system was based on a software package named "E-Groupware". This package was replaced with a similar package called Xoops because the new package contains more features, a much improved user interface, a larger support community, and was written in the same programming language, allowing code written for the old software to be easily ported into the new system. The EPS is now the default mechanism for users to submit their funding pre-proposals and associated paperwork. The system is also used by pre-proposal reviewers to access these documents during the pre-proposal review process. (Form FPR07) 2

24 PI: Anderson, Olin ARS Agreement #: NA As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (1): The EPS streamlines the pre-proposal process by making it more convenient for users to both submit and to review pre-proposals. The web interface is available 24 hours a day 7 days a week so users can perform their tasks at their convenience. The system is also faster than the old paper system which required forms be mailed to the Initiative's office. The process was also designed around the principle of making the whole submission process simple for the users and reviewers. Accomplishment (2): Electronic Document Management System Impact (2): The Document Management System (DMS) provides an easy to use interface for users to store electronic documents of any type in a secure and centralized location. This has helped the community to increase data sharing and collaboration. The DMS is a plugin module for the Xoops software package that is the basis for the EPS described above. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (2): The DMS allows for more community based information sharing and collaboration. Users have a simple interface that allows them to easily manage documents. They control their own online file systems and may keep documents private or share with them with any number of user/groups that have been registered in the Xoops system. Files may also be edited by users with proper permissions and then resubmitted to the DMS which has a comprehensive revision control system. (Form FPR07) 3

25 PI: Anderson, Olin ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Forum Proceedings: Canty, S., A. Clark, D. Ellis and D. Van Sanford (Eds.) Proceedings from the 2007 National Fusarium Head Blight Forum, Kansas City, MO; December 2-4. University of Kentucky, Lexington, KY. Proceedings are available in PDF format on the USWBSI s Website ( Newsletters: Lilleboe, D., S. Canty, and D. Van Sanford. Spring Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 8, Issue 1. Lilleboe, D., S. Canty, and D. Van Sanford. Fall Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 7, Issue 2. Press Releases: News Release: Scab Incidence Varies Across Midwestern States News Release: Early Southern Scab Reports Mixed News Release: USWBSI Announces new Co-Chair News Release: Scab Disease Generally Low across U.S. in Press Release: USWBSI Mourns Loss of Co-Chair Tom Anderson. All Press Releases are available in PDF format on the USWBSI s Website ( (Form FPR07) 4

26 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Stephen Baenziger Institution: University of Nebraska Address: Department of Agronomy 330 Keim Hall Lincoln, NE Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 46,378 Developing Winter Wheat with Improved Fusarium Head Blight Tolerance by Conventional and Transgenic Approaches. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount To Enhance Variety Development of Scab Resistant Hard Winter $46,378 Wheat Varieties. Total Award Amount $ 46,378 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

27 PI: Baenziger, Stephen USDA-ARS Agreement #: Project 1: To Enhance Variety Development of Scab Resistant Hard Winter Wheat Varieties. 1. What major problem or issue is being resolved and how are you resolving it? The major problem that we are addressing is the susceptibility of wheat cultivars grown in Nebraska and adjacent regions. We are resolving this problem through: 1. developing elite hard winter wheat cultivars that are resistant to Fusarium head blight (FHB, scab) and 2. screening experimental lines in regional FHB nurseries and hard winter wheat regional nurseries to identify the level of FHB resistance within the existing elite winter germplasm of the Great Plains. In our crossing blocks we made 221 crosses specifically for FHB tolerance. The crossing effort was greatly aided by our receiving 255 backcrossed derived lines using MAS for the FHB1 allele developed in Wesley (an excellent variety and parent, released by the USDA-University of Nebraska cooperative breeding program and currently grown on 7.7% of Nebraska s acreage, field season statistics, NASS), Trego, and Harding from Dr. Guihua Bai. It should be noted that Wesley is predominantly grown in eastern NE and under irrigation, so its acreage in the primary region for FHB is larger than the reported 7.7%. A brief summary of the breeding progress so far is: Native or Marker Parent Marker Based Comment Crosses made 221 Using native resistance or known FHB QTLs F2 Populations 153 Using native resistance or known FHB QTLs F3 Populations 103 Using native resistance or known FHB QTLs F4 Headrows 3500 Using native resistance or known FHB QTLs F5 Lines Selected >100 57* Generation where line are tested with markers for FHB QTLs F6 Lines Selected Many of the 14 lines are agronomically unacceptable F7 Lines Selected NA** 1 F8 - F12 Lines Selected 14 0 Three years of FHB mist testing, mostly using native resistance Lines released (2008) 2 Native resistance *Have parents with FHB1 ** Second year of testing in Mist Nursery, hence waiting additional testing before being considered tolerant. We increased our crosses because we have better germplasm as parents, but also to help assist the SDSU winter wheat breeding program which is currently looking for a new winter wheat breeder (hence may not be making as many crosses as normal). We are not satisfied with the frequency of known QTLs in our breeding program so we are working with Dr. Bai to develop a F 1 half seed system for 3-way crosses and F 2 s after they have been sorted by Dr. Dowell (see below) to remove soft kernel types. Allele enrichment will be a critical strategy for the future. In addition, we have grown a replicated experiment in two locations to determine if our seed sorting efforts with Dr. Floyd Dowell is selecting for heritable traits. It appears that kernel hardness (critical as we use VA and MO soft winter wheat lines as parents) and color selection is heritable (it is doubtful that white wheats will be grown in the major FHB areas of the Great Plains). Sorting on protein content is still being studied but may be less heritable as might be expected for a trait with considerable environmental variation. Efficacy for sorting on low DON concentration is being evaluated in the field this year. The FHB nurseries were successfully screened and submitted for DON testing. (Form FPR07) 2

28 PI: Baenziger, Stephen USDA-ARS Agreement #: List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Two of the difficulties in the Hard Winter Wheat Region (HWWR) are that many lines are susceptible to FHB and that lines are poorly characterized, thus not allowing growers to select more tolerant lines for their farms (make educated decisions). In 2008, we released two new hard red winter wheat lines (e.g. Camelot (formerly NE01604) and Settler CL (formerly NH03614 CL) which have a higher level of tolerance to scab. Both lines are under seed increase and will be sold to certified seed producers in the fall, Camelot is a very high quality line for milling and baking and has an excellent disease resistance package. Settler CL is a new, broadly adapted herbicide tolerant wheat and will be co-released with South Dakota and Wyoming. Both of these wheats will complement Overland (formerly NE01643), our first release (in 2007) with better levels of FHB tolerance which will be widely available to growers in 2008 (expect a minimum of 120,000 units of seed to be available). Though these lines only give a partial improvement in reducing FHB, they are moving us away from truly susceptible lines. The three wheats can be grown with early flowering winter wheat as a way for growers to use flowering date to buffer against episodic events of FHB infection. Impact: We expect these three cultivars (Camelot, Settler CL, and Overland) will be widely grown and provide growers with three better choices for producing grain with lower DON on their farms. Overland is a superior agronomic line with acceptable end-use quality and stem rust resistance. Camelot has good agronomic performance, but better end-use quality and stem rust resistance than Overland. Settler CL has good end-use quality and stem rust resistance with the highest grain yield potential of the current herbicide tolerant wheats in NE and SD. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The growers of NE and SD will have agronomically superior cultivars that fit their cropping systems and that will lessen their risk to FHB. The millers and bakers of the hard winter wheat region will have grain that will have lower levels of DON, thus increasing food safety. We expect the tolerance to FHB in these cultivars will lessen the need for fungicides and will also increase the efficacy of fungicides when applied to reduce DON. (Form FPR07) 3

29 PI: Baenziger, Stephen USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Baenziger, P.S., B. Beecher, R.A. Graybosch, A. M. H. Ibrahim, D.D. Baltensperger, L.A. Nelson, Y. Jin, S. N. Wegulo, J.E. Watkins, J. H. Hatchett, Ming-Shun Chen, and Guihua Bai Registration of 'NE01643' wheat. J. Plant Registrations 2: Bockus, W. W., Baenziger, P. S., and Ibrahim, A. M. H Reaction of Kansas, Nebraska, and South Dakota winter wheat accessions to Fusarium head blight (FHB), Plant Disease Management Reports (online). Report 1:CF009. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. (Form FPR07) 4

30 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Guighua Bai Institution: USDA-ARS Address: 4008Throckmorton Hall Manhattan, KS Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 55,520 Marker-Assisted Characterization and Deployment of FHB- Resistance Genes in Wheat. USWBSI Individual Project(s) USWBSI Research Area * HGG HGR ARS Adjusted Award Project Title Amount Characterization of Novel QTLs for FHB Resistance in Asian Wheat Cultivars. $30,520 Improve FHB Resistance in Hard Winter Wheat by High-throughput Marker-assisted Selection. $ 25,000 Total Award Amount $ 55,520 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

31 PI: Bai, Guighua USDA-ARS Agreement #: NA Project 1: Characterization of Novel QTLs for FHB Resistance in Asian Wheat Cultivars. 1. What major problem or issue is being resolved and how are you resolving it? Effective utilization of FHB resistant resources relies on identification of new sources of resistance and understanding inheritance of their resistance to FHB and to DON accumulation in wheat grain. A major FHB resistance QTL from Sumai 3 has been mapped and widely used in breeding programs. Identification of resistance genes from new sources may enrich FHB resistance gene diversity and provide new sources of resistance genes to enhance FHB resistance levels through gene pyramiding. We identified new resistant sources that might be different from Sumai 3 after screening a collection of Asian wheat cultivars and landraces. We mapped scab resistance QTLs in Chinese landrace Wangshuibai and Chokwang from Korea. We are testing mapping populations with new Chinese landraces Haiyanzhong and Huangfangzhu as resistant parents in which the QTLs have not been mapped previously and markers for 3BS QTL showed different haplotypes from that of Sumai 3.We will elucidate genetic effects of these QTLs by testing the mapping populations for FHB resistance and DON content under greenhouse and field conditions. The results are also expected to provide breeders with quality markers for breeding wheat cultivars with low DON and high levels of FHB resistance to speed up breeding process. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: 1. The population of 250 F 6:7 RILs from the cross of Clark/Chokwang were evaluated in the greenhouse and field at Manhattan KS, spring 2008 for QTL validation. Data will be summarized and a manuscript will be prepared for publication. 2. About 140 F6 RILs from Wangshuibai/Wheaton were evaluated for field resistance in Manhattan, KS and Lincoln NE to investigate interaction between QTL and environments. 3. Two F5 RIL populations from the crosses between two Chinese landraces, Haiyanzhong and Huangfangzhu, and Wheaton (a highly susceptible parent) were evaluated for FHB resistance in a greenhouse experiment in Manhattan, KS. Marker screening work is initiated. 4. A high density map with about 900 SFPs derived from Affymetrix array and 300 SSRs was constructed using population Ning 7840/Clark. This is the first high density SFP map in wheat. A manuscript is prepared and will be submitted soon. Impact: Wangshuibai is a highly resistant landrace from China and Chokwang is a Korean cultivar with FHB resistance. The FHB resistance QTL from both QTLs are analyzed and compared with previously reported QTLs from Sumai 3 etc. The knowledge of similarity and difference of QTLs between these cultivars and Sumai 3 will help breeders to select right resistant parents and develop new breeding strategy for pyramiding FHB resistance QTLs from diverse origins. The SFP map developed will be a powerful tool for high resolution (Form FPR07) 2

32 PI: Bai, Guihua ARS Agreement #: NA FY04 Final Performance Report mapping of wheat resistance QTL and developing SNP markers for high-throughput genotyping. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The information on QTLs from different sources and linked molecular markers will be published in a peer-reviewed journal and represents an important technology transfer event because wheat geneticists, pathologists and breeders will have access to QTL and markers in their research or breeding programs. Also, the SFP map was the first high density SNP map developed from a FHB mapping population and will be used by FHB researchers worldwide for FHB mapping and marker identification. Project 2: Improve FHB Resistance in Hard Winter Wheat by High-throughput Marker-assisted Selection. 1. What major problem or issue is being resolved and how are you resolving it? The best source of FHB resistance in wheat is mainly spring type that derived from Sumai 3 from China. Hard winter wheat cultivars grown in the U.S. are usually highly susceptible to FHB. Progress in breeding HWW for improved FHB resistance has been slow. Major FHBresistance QTL from Sumai 3 has been mapped and closely linked markers were identified. This QTL has been widely used in spring wheat or soft winter wheat breeding programs in the U.S. with great success. However, it has not been deployed in commercial hard winter wheat cultivars yet. To improve FHB resistance in hard winter wheat, we use marker-assisted backcross method to transfer the 3BS major QTL as well other minor QTLs into adapted hard winter wheat backgrounds. The results are expected to provide breeders with hard winter wheat lines that have low DON content and a high level of FHB resistance. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The major QTL on 3BS from Sumai 3 and its derivatives has been transferred into hard white winter wheat cultivar Trego from Kansas, and hard red winter wheat Wesley from NE and Harding from South Dakota. The 200 Bc2F4 lines carrying the 3BS QTL were selected and continuously evaluated for FHB resistance and other agronomic traits in the field experiments at Manhattan KS and Lincoln, NE. The result indicated that most selected lines have good winter hardiness and similar agronomic traits as recurrent parents but better FHB resistance than that of recurrent parents under severe FHB epidemics in greenhouse experiment and field experiments of 2007 and Five to ten best lines will be selected from each back ground and released as breeding parents to HWW breeding programs. Meanwhile, a new backcross was developed between (Ning7840/Jagger)F1 and (Jagger /Chokwang)F1 to pyramiding different QTLs. Its F1 carrying 3-4 known QTL from two (Form FPR04) 3

33 PI: Bai, Guihua ARS Agreement #: NA FY04 Final Performance Report resistant parents were selected using markers and backcrossed or crossed with locally adapted Jagger and Overlay from Kansas and Overland from Nebraska. Impact: FHB has been severe in many fields of KS and NE this year and current used HWW cultivars grown in the Great Plains and elite breeding lines available are highly susceptible to FHB. Marker analysis didn t detect 3BS QTL in the breeding lines from HWW regional nurseries. This is the first time to use marker-assisted backcross to transfer 3BS resistance QTL into US hard winter wheat. The resulting breeding lines will be used as breeding parents to speed up FHB improvement in hard winter wheat and these materials may also be released as FHB resistant cultivars after further field selection and FHB evaluation. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Current elite breeding lines from HWW regional nurseries don t carry the 3BS major QTL FHB resistance. After the project is accomplished, breeders from Kansas, South Dakota and Nebraska will have adapted wheat breeding materials that carry 3BS major QTL for FHB resistance. (Form FPR04) 4

34 PI: Bai, Guighua USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Yu GH, Ma HX, Bai GH, Tang KX SSCP markers associated with a major QTL for Fusarium head blight resistance in wheat. Mol. Biology (accepted) 2. Yu J-B, Bai G-H, Cai S-B, Dong Y-H, and Ban T New FHB-resistant Sources from Asian Wheat Germplasm. Crop Sci. (in press). 3. Yu J-B, Bai G-H, Zhou W-C, Dong Y-H and Kolb FL Quantitative trait loci for Fusarium head blight resistance in recombinant inbred population Wangshuibai/Wheaton. Phytopathology 98:87-94 Meeting Abstract: 1. A.N. Bernardo, S-W. Hu, P.J. Bradbury, R.L. Bowden, E.S. Buckler and G-H. Bai Using Affymetrix array to discover single nucleotide polymorphisms in wheat Genome XVI, January 12-16, 2008 San Diego, CA. 2. A.N. Bernardo, S-W. Hu, P.J. Bradbury, R.L. Bowden, E.S. Buckler and G-H. Bai Using Affymetrix array to discover single nucleotide polymorphisms in wheat National Fusarium Head Blight Forum, December 2-4, Kansas City, KS. 3. G Bai, P St. Amand, D Zhang, A Ibrahim, SP Baenziger, B Bockus, and A Fritz Marker-assisted Transfer of 3BS QTL for FHB Resistance into Hard Winter Wheat National Fusarium Head Blight Forum, December 2-4, Kansas City, KS. 4. A.N. Bernardo, S-W. Hu, P.J. Bradbury, R.L. Bowden, E.S. Buckler and G-H. Bai Using Affymetrix array to discover single nucleotide polymorphisms in wheat. Wheat Genomics Conference Nov 31-Dec 2. Kansas City, KS. 5. G Bai, P St. Amand, D Zhang, A Ibrahim, SP Baenziger, B Bockus, and A Fritz Improvement of FHB Resistance of Hard Winter Wheat through Marker-assisted Backcross. ASA-CSSA-SSSA 2007 International Annual Meeting, Nov 4-8, New Orleans Louisiana (Form FPR04) 5

35 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Gary Bergstrom Institution: Cornell University Address: Department of Plant Pathology 334 Plant Science Building Ithaca, NY Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 60,488 Developing Technologies to Enhance Utility for B. Subtilis Against Wheat Scab. USWBSI Individual Project(s) USWBSI Research Area * CBCC EEDF ARS Adjusted Award Project Title Amount Evaluation of Integrated Management Strategies for FHB in New York. $20,488 Relative Contribution of within-field Inoculum to FHB Infection of Wheat. $ 40,000 Total Award Amount $ 60,488 7/14/2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

36 PI: Bergstrom, Gary USDA-ARS Agreement #: Project 1: Evaluation of Integrated Management Strategies for FHB in New York. 1. What major problem or issue is being resolved and how are you resolving it? This research in New York was part of a multi-state, multi-year field study aimed at demonstrating that integrated management is the most efficacious and economical approach to management of Fusarium head blight and DON. The individual and combined effects of cultivar resistance (four wheat cultivars varying in resistance to FHB) and fungicide (Prosaro at initiation of flowering) were assessed for their effects on FHB, DON, and grain yield. In New York we conducted two separate experiments in 2007, one in wheat planted no-till into corn stubble and one planted after conventional tillage following soybean harvest. Central New York experienced one of the driest May-July periods on record such that FHB symptoms were not observed in the plots. And yet, detectable levels of DON were recorded for some plots as a result of infection later during grain development. There were no significant effects of treatment except that the FHB susceptible cultivar Caledonia outyielded the other cultivars in the conventional tillage environment following soybean in the virtual absence of wheat fungal diseases. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We demonstrated that DON can result from late infections with no associated symptoms and that this late DON accumulation may not be reduced by fungicide application at initiation of flowering. Impact: The detection of DON in grain from plants that exhibited no visual symptoms of FHB is a reminder that successful integrated management must also address late infections resulting in DON. FHB management should not end at the initiation of flowering. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: It is unlikely that fungicide application past the initiation of flowering will be available as a solution to late infections that result in DON accumulation. But resistant cultivars and applications of biocontrol agents during grain formation should be assessed for extension of FHB management through the grain filling period. (Form FPR07) 2

37 PI: Bergstrom, Gary USDA-ARS Agreement #: Project 2: Relative Contribution of within-field Inoculum to FHB Infection of Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Knowledge of the relative contribution of within-field inoculum sources (i.e., corn and small grain crop debris) of Gibberella zeae to infection of local wheat is important for developing and/or excluding strategies for managing FHB. We utilized a marked isolate, releaserecapture approach to assess relative contribution of spores from corn stalks to infection of wheat heads at the corn stalk source and at 10 foot and 20 foot radii from the corn stalk sources as well as in more distant portions of the fields. The 2007 experiment was conducted in commercial scale wheat fields in New York and Virginia that lacked corn or cereal residues. Isolates of G. zeae from wheat heads were scored for AFLP haplotype and were categorized for identity or non-identity with released clonal isolates possessing unique AFLP haplotypes. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We provided the first direct (genetic) evidence of the relative contribution of local inoculum to local infection of wheat heads by Gibberella zeae. Both within-field, infected corn stalks and background atmospheric sources provided significant levels of inoculum for infection of local wheat heads in a non-epidemic environment in New York and in a moderate FHB epidemic environment in Virginia in Yet, in both cases, local corn stalk isolates of G. zeae contributed less than half of the spores resulting in wheat head infection immediately above the corn stalks and a small fraction of the spores resulting in head infection at 10 foot or greater distances from the corn stalks. A small number of isolates of the released clones were recovered in distant portions of the fields indicating a contribution of corn stalk-derived spores to the mixed atmospheric population of the fungus. Impact: The strong implication of these first year findings is that within-field corn or cereal debris contributes less spores for infection of local wheat or barley heads than does the regional atmospheric population of fungal spores. That is not to say that within-field sources are not important or that they should not be reduced as a component of integrated management of FHB. But our findings suggest strongly that debris management in single fields achieved through tillage, crop sequence, or the application of fungicides or biocontrol agents to stubble is unlikely, by itself, to result in large reductions of FHB or DON. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The wheat and barley community now has a more realistic idea of the relative contributions of within-field and area atmospheric sources of inoculum for FHB. The largest vulnerability is from regional populations of spores in the atmosphere. The localized benefits of debris management may be less than some have expected. Protection of cereals with resistant varieties, fungicides, and biocontrol agents should result in benefits whether spores come from within-field or regional sources. (Form FPR07) 3

38 PI: Bergstrom, Gary USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Publications (peer reviewed): Schmale, D.G. III and G.C. Bergstrom The aerobiology and population structure of Gibberella zeae. Plant Health Progress DOI: /PHP RV. Pryor, S.W., D.M. Gibson, G.C. Bergstrom, and L.P. Walker Minimization of betweenwell sample variance of antifungal activity using a high-throughput screening microplate bioassay. Biotechniques 42: Publications (not peer reviewed): Bergstrom, G.C., and D.G. Schmale Aerobiology of Gibberella zeae: Whence come the spores for Fusarium head blight? Pages in Proc National Fusarium Head Blight Forum, The Westin Crown Center, Kansas City, Missouri, December 2-4, Keller, M.D., K.D. Duttweiler, D.G. Schmale, and G.C. Bergstrom Contribution of within-field inoculum sources to Fusarium head blight in wheat. Page 98 in Proc National Fusarium Head Blight Forum, The Westin Crown Center, Kansas City, Missouri, December 2-4, Presentations for Scientific/Professional Audiences: Bergstrom, G.C. and D. G. Schmale. Aerobiology, regional epidemiology and population genetic structure of Gibberella zeae. 62 nd Northeastern Corn Improvement Conference, Boyce Thompson Institute, Cornell University, Ithaca, NY, February 15-16, 2007 Bergstrom, G.C., and D.G. Schmale. Aerobiology of Gibberella zeae: Whence come the spores for Fusarium head blight? 2007 National Fusarium Head Blight Forum, The Westin Crown Center, Kansas City, Missouri, December 2-4, Extension presentations by Gary C. Bergstrom in that included updates on Fusarium head blight research: Seed Growers Field Day. Ithaca, NY. Small grain and forage crop disease update; Fusarium head blight (ca. 65 persons) (7/8/08) Small Grains Management Field Day. Aurora, NY. Integrated management of Fusarium head blight. (ca. 100 persons) (6/5/08) North Country Crop Congress, Carthage, NY. Field crop disease update. (ca. 50 persons) (3/13/08) North Country Crop Congress, Madrid, NY. Field crop disease update. (ca. 50 persons) (3/12/08) (Form FPR07) 4

39 PI: Bergstrom, Gary USDA-ARS Agreement #: Monroe Tractor Spray Clinic. Auburn, NY. Plant disease update. (ca. 200 persons) (3/11/08) New York State Small Grains Seed Committee. Waterloo, NY. Cereal disease research and extension update. (12 persons) (3/4/08) Seneca County Crop and Dairy Day. Romulus, NY. Field crop disease update. (ca. 25 persons) (2/20/08) Madison County Crop Congress. Cazenovia, NY. Disease and mycotoxin threats to New York crops. (ca. 50 persons) (1/23/08) Cornell Nutrition Conference For Feed Manufacturers. East Syracuse, NY. Field practices that reduce mycotoxins. (ca. 400 persons) (10/25/07) Field Crop Dealer Meeting. Auburn, NY. An update on disease and mycotoxin threats to New York field crops. (ca. 60 persons) (10/26/07) Field Crop Dealer Meeting. Batavia, NY. An update on disease and mycotoxin threats to New York field crops. (ca. 60 persons) (10/25/07) Field Crop Dealer Meeting. New Hartford, NY. An update on disease and mycotoxin threats to New York field crops. (ca. 60 persons) (10/24/07) Field Crop Dealer Meeting. Clifton Park, NY. An update on disease and mycotoxin threats to New York field crops. (ca. 60 persons) (10/23/07) Seed Growers Field Day. Ithaca, NY. Field crop disease update. (ca. 50 persons) (7/5/05) Small Grains Management Field Day. Aurora, NY. (ca. 75 persons) (6/7/07) Vermont Crops n Critters Conference, Bridport, VT. Sustainable wheat health management for the Northeast. (ca. 50 persons) (3/20/07) Steuben County Crop Symposium, Bath, NY. Field crop disease update. (ca. 120 persons) (2/27/07) Finger Lakes Soybean and Small Grains Congress, Waterloo, NY. Best management practices for Fusarium head blight. (ca. 100 persons) (2/8/07) Western New York Soybean and Small Grains Congress, Batavia, NY. Best management practices for Fusarium head blight. (ca. 100 persons) (2/7/07) (Form FPR07) 5

40 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: William Berzonsky Institution: North Dakota State University Address: Department of Plant Sciences 370G Loftsgrad Hall Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 33,476 Hastening the Development of Specialty Spring Wheats with Resistance to Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * IIR ARS Adjusted Award Project Title Amount Pyramiding Host Genes to Reduce DON and Develop Scab $33,476 Resistant Spring Wheat. Total Award Amount $ 33,476 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

41 PI: Berzonsky, William USDA-ARS Agreement #: Project 1: Pyramiding Host Genes to Reduce DON and Develop Scab Resistant Spring Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Spring wheat genotypes which express a single major source of resistance to fusarium head blight (FHB) may not provide the maximum level of resistance or provide the greatest reduction in deoxynivalenol (DON). Hence, using molecular markers, we identified doubled-haploid lines and developed spring wheat breeding lines which combine or pyramid two different sources of FHB resistance. This project was designed to assess whether or not resistance to FHB has been enhanced as a result of combining the Sumai-3 source of FHB resistance with a source of resistance from Triticum dicoccoides. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Two related spring wheat lines, designated NDSW0714 and NDSW0715, were developed such that they contained one and two QTL for FHB resistance, respectively. In three separate greenhouse experiments, spikes of NDSW0714 and NDSW0715 were injected with Fusarium graminearum and compared for expression of resistance. Resistance to FHB was assessed by measuring disease severity, percent fusarium-damaged kernels (FDK), and DON content. Across all experiments, the mean disease severity rating of NDSW0715 at 21 days after inoculation was 8.3%, which was significantly lower than the 17.6% rating for NDSW0714. Mean FDK and DON content measurements of the lines were not significantly different. Impact: Few studies have examined the impact of pyramiding different genes with resistance to FHB into a single genotype. Since pyramiding genes for resistance from different sources and developing acceptable varieties is not necessarily an easily accomplished breeding task, particularly if at least one source derives from alien germplasm, it is important to determine if pyramiding enhances resistance. Results of this study indicate that there could be a significant benefit derived from pyramiding genes, which is demonstrated by a decrease in the level of disease severity over time. It provides information that will help breeders determine if pyramiding genes will be useful in developing host plant resistance to FHB, and it ultimately will result in the release of a spring wheat germplasm line which combines two different genes for resistance to FHB. It also demonstates that molecular markers can be effectively employed to pyramid different genes, despite these genes expressing a similar Type II phenotypic resistance to FHB. (Form FPR07) 2

42 PI: Berzonsky, William USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community now has a better understanding of how two different genes for resistance to FHB interact in the host wheat plant to enhance the level of resistance to FHB and potentially reduce the production of DON. The use of molecular markers to effectively pyramid different genes validates this approach for plant breeders who might be interested in combining genes that still express a similar type of phenotypic resistance. Seed of NDSW0715 is being increased with plans to release it as germplasm and both NDSW0714 and NDSW0715 are being tested in statewide and regional FHB nurseries. Once released, NDSW0715 will provide wheat breeders with potential parental material in which the Sumai-3 and Triticum dicoccoides sources of resistance to FHB have been packaged together in an adapted type, similar in phenotype to Alsen spring wheat. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Berzonsky, W.A., B.L. Gebhard, E. Gamotin, G.D. Leach, and S. Ali. A reciprocal backcross monosomic analysis of the scab resistant spring wheat (Triticum aestivum L.) cultivar, Frontana. Plant Breed. 126: Berzonsky, W.A., E.L. Gamotin, and G.D. Leach Enhancing host resistance to fusarium head blight: Pyramiding genes in spring wheat. ASA-CSSA-SSSA Annual Meetings - New Orleans, LA (Poster presentation). Berzonsky, W.A., E.L. Gamotin, G.D. Leach, and T Adhikari Enhancing host resistance to fusarium head blight: Pyramiding genes in spring wheat. USWBSI Annual Meetings - Kansas City, MO (Poster presentation). (Form FPR07) 3

43 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Bruce Bleakley Institution: South Dakota State University Address: Department of Biology & Microbiology NPBL-Box 2140D, Room 252 Brookings, SD Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 9,756 Management of Fusarium Head Blight with Biological Control Agents. USWBSI Individual Project(s) USWBSI Research Area * CBCC ARS Adjusted Award Project Title Amount Management of Fusarium Head Blight With Biological Control $9,756 Agents. Total Award Amount $ 9,756 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

44 PI: Bleakley, Bruce USDA-ARS Agreement #: Project 1: Management of Fusarium Head Blight With Biological Control Agents. 1. What major problem or issue is being resolved and how are you resolving it? a) A major part of the project is to quantify numbers of bacterial biocontrol agents (BCAs) (for our project, selected Bacillus spp.) after they are sprayed onto heads of wheat and barley. For the 2006 and 2007 growing seasons, we have counts of the BCA Bacillus 1BA. More than two years of BCA population data are needed to better understand how 1BA behaves in the field after spray application. --More data are sought by continuing to do similar BCA population counts via field plot work at Brookings, SD in the summer of 2008, to provide a larger data set to better gauge population fluctuations of 1BA after it is sprayed onto wheat and barley heads in the field. We are also involved in the Uniform Biological Control tests that are part of the Scab Initiative. b) There is still a need for evidence that 1BA and related BCAs produce metabolites such as lipopeptides (such as iturin and surfactin) on the grain heads. These lipopeptides are thought to be the major mechanism whereby 1BA and several other Bacillus spp. used as BCAs inhibit growth of F. graminearum, reduce FHB, and/or reduce DON levels. --Resolution of the problem is being sought by studies done over the last year, and also in progress with Chris Dunlap of USDA-ARS-Peoria, analyzing methanol extracts from grain heads via mass spectrometry (MALDI-TOF) to semi-quantitatively assay the amount of lipopeptide present. c) There is also a need for evidence of lipopeptide genes on treated grain heads using PCR. Resolution of this is sought by ongoing studies that extract DNA from inoculated grain heads, then use PCR to verify presence of lipopeptide genes in the sample. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: a) Field plot population studies of 1BA: Populations of Bacillus strain 1BA were studied during 2006 and 2007 in field plots at Brookings, SD after spray application at anthesis. Field plot treatments in 2006 used cells of 1BA grown in several different broth media; and including the fungicide Folicur with a spreader or sticker in some treatments. Field plot treatments in 2007 used only tryptic soy broth cultures of 1BA by itself; and 1BA + Prosaro (a fungicide mix) + Induce NIS (nonionic surfactant). Population counts were done over a 20 day period via MPN technique. Controls not receiving 1BA inoculation gave low cell counts in both years, no higher than about 3.5 X 10 2 CFU/g fresh plant weight. Numbers on barley heads were low, comparable to control treatments, throughout the 2007 study. In both years, there was a rapid decline in numbers of 1BA on wheat heads in the first few days after spraying, followed by an increase in numbers in some treatments to levels around 10 4 CFU/g fresh plant weight. Treatments using 1BA grown in defined broth yielded lower numbers of 1BA on wheat heads over time than treatments using 1BA grown in complex broth. In both years, some treatments showed a major increase in 1BA numbers around 10 days after spraying. In 2007 in the treatment with 1BA alone, vegetative cell counts peaked around day 10, then declined sharply, followed by a second increase in numbers by day 20, with an increase in endospore numbers. Compared to the treatment with 1BA alone, in the treatment combining 1BA with Prosaro and Induce NIS, population peaks shifted in time. Peak numbers of vegetative cells occurred at about (Form FPR07) 2

45 PI: Bleakley, Bruce USDA-ARS Agreement #: day 6 then declined. As vegetative numbers of 1BA in this treatment declined, endospore numbers increased. Impact: Two years of field plot data showed similar behavior of 1BA after spray application onto wheat heads at anthesis. This work documents that our Bacillus strain 1BA persists and grows on wheat heads after application, which is considered to be an important trait for a BCA that controls a pathogen by producing antibiotics such as lipopeptides. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: About 10 days after wheat anthesis, there appears to be a predictable increase in numbers of 1BA sprayed onto heads at anthesis. Survival and growth of the BCA on wheat heads is greatly affected by the medium used to grow BCA inoculum, and by fungicides and adjuvants in spray. Population fluctuations of 1BA on barley heads are different from that on wheat heads. Accomplishment: b) Evidence of lipopeptide in wheat heads after spraying: Methanol extracts of wheat heads inoculated with 1BA were processed and sent to Chris Dunlap s laboratory. Initial results were negative for presence of lipopeptide. Dunlap s analytical protocol works well with Bacillus broth populations of 10 7 CFU/ml or higher; whereas our samples from inoculated wheat heads were shown by MPN counts to have 1BA numbers of about 10 4 CFU/g. We are working with Dunlap to establish the lower limit of detection for numbers of lipopeptide-producing Bacillus his technique can detect. Impact: Establishing a lower limit of detection for number of lipopeptide-producing Bacillus that can be detected by MALDI-TOF analysis is being sought, to see if cell numbers lower than that in dense broth cultures can be detected. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? Lower limit of detection for numbers of lipopeptide-producing BCAs on plant material is not certain, but will become more certain after this work proceeds further. Accomplishment: c) Evidence of lipopeptide genes on inoculated grain heads using PCR: DNA extracted from pure cultures of 1BA in the laboratory show amplified bands after PCR that indicate presence of lipopeptide genes. Work later this year will focus on using the same methodology to look for similar bands in amplified DNA extracted from inoculated grain heads. Impact: Pure culture work with strain 1BA verifies that it has the expected bands indicative of lipopeptide genes. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Detection of lipopeptide genes in DNA extracted from grain heads treated with lipopeptideproducing BCAs should be possible, once PCR parameters are optimized. (Form FPR07) 3

46 PI: Bleakley, Bruce USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Morgan, J., B.H. Bleakley, and C.A. Dunlap Field and laboratory studies to monitor cell populations, lipopeptides and lipopeptide genes of Bacillus 1BA, a biocontrol agent active against Fusarium Head Blight. Abstract, pg In: S.M. Canty, A. Clark, D. Ellis, and D. Van Sanford (eds.), Proceedings of the 2007 National Fusarium Head Blight Forum, Dec. 2-4, 2007, Kansas City, Missouri. East Lansing; Michigan State University. Ruden, K.R., B.H. Bleakley, and B.E. Ruden Uniform trials for the performance of biological control agents in the suppression of Fusarium Head Blight in South Dakota. Abstract, pg In: S.M. Canty, A. Clark, D. Ellis, and D. Van Sanford (eds.), Proceedings of the 2007 National Fusarium Head Blight Forum, Dec. 2-4, 2007, Kansas City, Missouri. East Lansing; Michigan State University. Yuen, G.Y., C.C. Jochum, S. Halley, G. Van Ee, V. Hoffman, and B.H. Bleakley Effects of spray application methods on biocontrol agent viability. Pp In: S.M. Canty, A. Clark, D. Ellis, and D. Van Sanford (eds.), Proceedings of the 2007 National Fusarium Head Blight Forum, Dec. 2-4, 2007, Kansas City, Missouri. East Lansing; Michigan State University. Yuen, G.Y., C.C. Jochum, K.R. Ruden, J. Morgan, B.H. Bleakley, and L.E. Sweets Results from the 2007 standardized evaluation of biological agents for the control of Fusarium Head Blight on wheat and barley. Pp In: S.M. Canty, A. Clark, D. Ellis, and D. Van Sanford (eds.), Proceedings of the 2007 National Fusarium Head Blight Forum, Dec. 2-4, 2007, Kansas City, Missouri. East Lansing; Michigan State University. Bleakley, B.H., and J. Morgan Field Plot Studies of Survival and Growth of the Biocontrol Agent Bacillus Strain 1BA Applied To Wheat Heads. Poster abstract: In Abstracts of the 108 th General Meeting of the American Society for Microbiology; June 1-5, 2008; Boston, MA. Baye, N.L Bacillus strains used for biological control of Fusarium Head Blight: identification, growth studies, and lipopeptide production. Master thesis, South Dakota State University, Brookings, SD. (Form FPR07) 4

47 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Ann Blechl Institution: USDA-ARS Address: Western Regional Research Center 800 Buchanan St. Albany, CA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 7,000 Wheat Transformants Containing DON Resistant Versions of Wheat L3 Ribosomal Protein. USWBSI Individual Project(s) USWBSI Research Area * GET ARS Adjusted Award Project Title Amount Modification of the Ribosomal Target to Enhance Resistance to $7,000 Trichothecene Mycotoxins. Total Award Amount $ 7,000 7/14/2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

48 PI: Blechl, Ann USDA-ARS Agreement #: NA Project 1: Modification of the Ribosomal Target to Enhance Resistance to Trichothecene Mycotoxins. 1. What major problem or issue is being resolved and how are you resolving it? One of the most cost-effective ways to prevent losses due to Fusarium Head Blight (FHB) is host plant resistance. Although applications of traditional breeding have resulted in introgression of Type II resistances into wheat, these are only partial and can be overcome by large inocula of Fusarium spores. The deoxynivalenol (DON) toxin produced by Fusarium species acts as a virulence factor in increasing the success of fungal colonization and spread. The target of the toxin is the ribosomal protein L3. Genetic transformation can be used to introduce new sources of resistance into wheat. In previous work with collaborators Nilgun Tumer and Di Rong (Rutgers University, New Brunswick, New Jersey), we have shown that transgenic wheat seedlings with high levels of a truncated form of the yeast L3 protein are protected against DON toxicity. More recently, Tumer and colleagues isolated genes encoding wheat L3 ribosomal proteins. They prepared DNA constructions fusing the highly expressed maize Ubiquitin1 promoter and first intron to two different DON-resistant mutants of the wheat RPL3A1 gene and to a truncated wheat RPL3A1 gene that encodes the L3 protein fragment. Each coding region included V5 and 6xHis epitope tags so that the new proteins can be distinguished from native wheat L3. The work for the project that is the subject of this report was to introduce these three constructs into wheat using particle bombardment. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Wheat transformants have been identified that contain the constructs for each DON-resistant mutant form of the wheat L3 gene or for the wheat L3 gene fragment. Inheritance to the T 1 generation has been confirmed for 9 transformants containing the W258C (cysteine for tryptophan at position 258) mutant form of the wheat gene, 12 transformants containing the W258K (lysine for tryptophan) mutant form of the wheat gene, and 4 transformants containing the truncated wheat gene encoding the L3 fragment. Some putative transformants still need to be characterized for inheritance. Impact: After securing interstate movement permits from APHIS, T 1 or T 2 seed from the transformants will be sent to Rutgers University where Tumer and colleagues will characterize expression of the L3 mutant and L3 fragment genes in various tissues. The resistance of transgene-expressing wheat seedlings to DON will be determined. If toxin resistance is demonstrated, selected lines will be greenhouse- and field-tested for DON accumulation levels and resistance to FHB. If this research is successful, these transgene loci will provide genetically marked sources of novel resistance to FHB. (Form FPR07) 2

49 PI: Blechl, Ann USDA-ARS Agreement #: NA As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community will have a new and independent way of attaining resistance to FHB. This strategy utilizes over-expression of variant forms of natural wheat proteins and thus may be more acceptable to the public than introduction of completely alien proteins into wheat. As barriers to introduction of genetically engineered wheat into commercial production are overcome, wheat breeders can combine this type of resistance with Type I and Type II resistances from other sources to provide producers with wheat cultivars with robust multi-genic FHB resistance. (Form FPR07) 3

50 PI: Blechl, Ann USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. NONE (Form FPR07) 4

51 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Sue Blodgett Institution: South Dakota State University Address: Plant Science Department Rm 219 Ag Hall Brookings, SD USA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 22,699 Field Studies on Chemical and Biological Control of Fusarium Head Blight in South Dakota. USWBSI Individual Project(s) USWBSI Research Area * CBCC CBCC ARS Adjusted Award Project Title Amount Collaboration on Uniform Fungicide and Biological Control Trials in South Dakota. $12,864 Evaluation of Integrated Management Strategies for Fusarium Head Blight in SD. $ 9,835 Total Award Amount $ 22,699 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

52 PI: Blodgett, Sue USDA-ARS Agreement #: Project 1: Collaboration on Uniform Fungicide and Biological Control Trials in South Dakota. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight can only be managed by a combination of cultural and chemical means. In many cases, producers have to compromise on cultural approaches at disease management and become reliant on chemical control. Fusarium head blight is an erratic problem in SD, but was severe over a wide area of the state in 2005; however, the impact of Fusarium head blight was significantly reduced in 2007 due to statewide drought. The continued availability of tebuconazole (Folicur, Orius, Embrace, and TebuStar) via Section 18 has become widely accepted among growers and has become a common production input in some parts of the state. Full registration of a new product, Bayer s Proline (prothioconazole) was given in early Availability of the Section 18 fungicides plus having a new Section 3 fungicide gave the producers additional options for spraying wheat and barley for Fusarium head blight in More effective fungicides or application methods are needed to provide better disease control and improve the profit margin of producers. Awareness of FHB risk has been elevated and is a significant consideration in grower decisions. We have continued to screen products through the uniform fungicide trial, participated in the mini -uniform biological trial, and have also screened SDSU biological products in the field for Dr. Bruce Bleakley. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Success in using the uniform trials to show producers that fungicides can make a difference in FHB suppression. The identified products tebuconazole, metconazole, and prothioconazole continue to be the most efficacious fungicide chemistries. Impact: Full registration of a new product, Bayer s Proline (prothioconazole) was given in early This finally puts the tools in the hands of the producer without the need for special labels. In early 2008, EPA has granted full registrations for the following products: Bayer s Folicur (tebuconazole), Mana s Orius (tebuconazole) and BASF s Caramba (metconazole). These registrations have helped the producers by allowing them to have various options for fungicide use against Fusarium head blight. Having additional pesticides available should reduce shortages as a whole, as several of these chemicals were in short supply in As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: In 2008, the producer has more choices of efficacious products than they have had in previous years and with special labels. (Form FPR07) 2

53 PI: Blodgett, Sue USDA-ARS Agreement #: Project 2: Evaluation of Integrated Management Strategies for Fusarium Head Blight in SD. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight can only be managed by a combination of cultural and chemical means. In many cases, producers have to compromise on cultural approaches at disease management and become reliant on chemical control. Fusarium head blight is occasionally a severe problem in the state (2005); however, damage was significantly reduced in 2007 due to statewide drought. Integrated methods are needed to provide better disease control and improve the profit margin of producers. Awareness of FHB risk has been elevated and is a significant consideration in grower decisions. Plant breeding to introduce increased tolerance and resistance to Fusarium head blight continues to move forward. We are now cooperating with seven other states (Ohio, North Dakota, Kentucky, Missouri, Nebraska, New York and Kansas) to evaluate the effects of multiple strategies including chemical, crop rotation and varietal selections for Fusarium head blight and DON control under different environments. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Success in using an integrated management strategy so far has shown that using a moderately resistant variety paired with fungicide control shows the highest percent control of Fusarium head blight. When using different cropping scenarios, the one management scheme to use to get the highest control of Fusarium head blight is to use a non-host crop plus a moderate resistant variety plus a fungicide treatment. This study is helping to show producers that making the right choices in their cropping systems, selecting appropriate varieties plus fungicides can make a difference in FHB suppression. Impact: Increasing producer knowledge of the effects of cropping systems, understanding the disease cycle and what tools they can use to develop an integrated management program to prevent/suppress Fusarium head blight. This approach may ultimately reduce the use of fungicides by incorporating a moderately resistant variety paired with a non-host rotational crop to reduce inoculum load for Fusarium head blight in their wheat or barley. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: So far this study has shown that what has been the general idea about different cropping systems paired with varieties that have different resistance levels do have a difference in the levels of Fusarium head blight and that using a moderately resistant variety paired with a non-host crop and a fungicide spray will lessen the severity of Fusarium head blight in the years that are conducive to having Fusarium head blight. (Form FPR07) 3

54 PI: Blodgett, Sue USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Paul, P., L. Madden, M. McMullen, D. Hershman, L. Sweets, S. Wegulo, W. Bockus, S. Halley and K. Ruden An integrated approach to managing FHB and DON in Wheat: Uniform Trials National Head Blight Forum. December 2-4, 2007, Kansas City, MO, Proceedings, pp Paul, P., L. Madden, M. McMullen, D. Hershman, D. Brown-Rytlewski, L. Sweets, E. Adee, C. Bradley, B. Padgett and K. Ruden Fungicide effects on FHB and DON in wheat class across multiple locations and wheat classes: uniform fungicide trials National Head Blight Forum. December 2-4, 2007, Kansas City, MO, Proceedings, pp Ruden, K.R. and B.E. Ruden Field Plot Summaries: Plant Disease and Fungicide Trials. SDSU Extension Service. 54 p. online at Ruden, K.R., B.E. Ruden, K.D. Glover, and J.L. Kleinjan uniform fungicide performance trials for the suppression of Fusarium head blight in South Dakota. National Head Blight Forum. December 2-4, 2007, Kansas City, MO, Proceedings, p Ruden, K.R., B.H. Bleakley, and B.E. Ruden uniform trials for the performance of biological control agents in the suppression of Fusarium head blight in South Dakota. National Head Blight Forum. December 2-4, 2007, Kansas City, MO, Proceedings, p Yuen, G.Y., C.C. Jochum, K.R. Ruden, J. Morgan, B.H. Bleakley, and L.E. Sweets Results from the 2007 standardized evaluation of biological agents for the control of Fusarium head blight on wheat and barley. National Head Blight Forum. December 2-4, 2007, Kansas City, MO, Proceedings, pp Presentations Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. January 4, Aberdeen, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. January 8, Faulkton and Selby, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. January 9, Timber Lake and Bison, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. January 10, Wall, SD. (Form FPR07) 4

55 PI: Blodgett, Sue USDA-ARS Agreement #: Ruden, B.E Fungicide Management in Small Grains What Happened to my Wheat? Crops Clinic. January 11, Martin, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Winter Wheat Meeting. January 15, Tripp County, SD. Ruden, B.E Disease Management for Season long disease protection Seed Treatment to Fungicide Timing. Ag Expo, SD Commercial Pesticide Applicators Training. January 17, Sioux Falls, SD. Ruden, B.E Disease Management for Season long disease protection Seed Treatment to Fungicide Timing. SD Commercial Pesticide Applicators Training. January 22 January 30, Rapid City, Pierre, Mitchell, Watertown, Sioux Falls, Yankton and Aberdeen, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. February 5, Bristol and Britton, SD. Ruden, B.E What Happened to my Wheat? Disease Management for Crops Clinic. February 5, Bristol and Britton, SD. Ruden, B.E What Happened to my Wheat? Disease Management for SD Private Applicators Training. February 6, Murdo, SD. Ruden, B.E What Happened to my Wheat? Disease Management for SD Private Applicators Training. February 6, Winner, SD. Ruden, B.E Disease Management for Season long disease protection Seed Treatment to Fungicide Timing. IPM Conference. February 8, Brookings, SD. Ruden, B.E Disease Management for Season long disease protection Seed Treatment to Fungicide Timing. SD Commercial Pesticide Applicators Training. February 11, Brookings, SD. Ruden, B.E What Happened to my Wheat? Disease Management Industry Day/Winfield Solutions Meeting. February 13, Brookings, SD. Ruden, B.E What Happened to my Wheat and Corn? Disease Management for Crops Clinic. February 14, Roscoe and Mellette, SD. Ruden, B.E Disease Management Timing, What s New, What s Changed. Crops Clinic. February 19, Pierre, SD. Ruden, B.E What Happened to my Wheat and Corn? Disease Management for Crops Clinic. February 26, Huron, SD. (Form FPR07) 5

56 PI: Blodgett, Sue USDA-ARS Agreement #: Ruden, B.E Hitting the Fungicide Target in Precision Placement through Application. Potter County Implement Crops Clinic. March 4, Gettysburg, SD. Ruden, B.E Disease Management Timing, What s New, What s Changed. Canby s Ag Crops Clinic. March 6, Ipswich, SD. Ruden, B.E Wheat Management SD Cooperative Agronomy Educator s Meeting. March 18, Philip and Pierre, SD. Ruden, B.E Disease Management Timing, What s New, What s Changed. Crop Consultant s Meeting. March 24, Brookings, SD. (Form FPR07) 6

57 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 14, 2008 Cover Page PI: Sue Blodgett Institution: South Dakota State University Address: Plant Science Department Rm 219 Ag Hall Brookings, SD USA Phone: Fax: Fiscal Year: 2007 FY07 ARS Agreement ID: Agreement Title: Winter Wheat Breeding for Scab Resistance in South Dakota. FY07 ARS Award Amount: $ 52,257 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount VDUN Winter Wheat Breeding for Scab Resistance in South Dakota. $52,257 Total Award Amount $ 52,257 Principal Investigator Date * BIO Biotechnology CBC Chemical & Biological Control EDM Epidemiology & Disease Management FSTU Food Safety, Toxicology, & Utilization GIE Germplasm Introduction & Enhancement VDUN Variety Development & Uniform Nurseries (Form FPR07)

58 PI: Blodgett, Sue USDA-ARS Agreement #: Project 1: Winter Wheat Breeding for Scab Resistance in South Dakota. 1. What major problem or issue is being resolved and how are you resolving it? Resistant varieties will be the main component of an integrated strategy to control scab. The development and implementation of resistant varieties is the most economical, sustainable, and long lasting means of control. We will continue to simultaneously select for resistance and desirable agronomic characteristics. Our long term objective is to continue to use traditional breeding techniques, aided by molecular marker selection (MAS), to develop and release FHB-resistant hard winter wheat varieties and germplasm with superior agronomic performance and end-use quality characteristics, excellent winter survival ability, and resistance to diseases prevalent in South Dakota and the northern Great Plains. Our specific objectives are to 1) use elite, FHBresistant germplasm with tagged QTLs, in addition to indigenous native resistant sources, in developing populations segregating for FHB resistance and desirable agronomic traits, 2) screen segregating populations, advanced lines, and established varieties in our mist-irrigated nursery and greenhouse for the purposes of line advancement and releasing and providing growers with accurate FHB ratings on commonly grown varieties, 3) use MAS as a complementary tool to select FHB-resistant lines, and 4) enter promising resistant lines into regional nurseries to facilitate development of varieties with broad adaptation in collaboration with the University of Nebraska and Kansas State University. We use mist-irrigated field screening nurseries to evaluate the material. Winter wheat would be vernalized in the early spring and then transplanted into the field in April. The program has successfully tested dormant seeding as an alternative to transplanting. All scab material is planted into 5 foot rows in the mist irrigated nursery while a transplanted nursery is used as a backup. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Darrell hard red winter wheat (HRWW) was released in It has the best FHB rating among all Great Plains HRWW varieties tested in South Dakota during the last six years. It ranked top for yield in South Dakota Crop Performance Testing (CPT) Variety Trial in 2006 and had an exceptional three-year yield average. It had exceptional performance in the state of Nebraska in the Northern Regional Performance Nursery (NRPN) in 2003 and It has acceptable milling, good baking quality, and a good diseases package. 1,500 lines were dormant seeded in November 2006 in the mistirrigated nursery in Brookings, SD. Due to excessive early spring rain, the seed failed to germinate and the nursery was lost for the second time in the last six years. A backup nursery consisting of 397 lines, including the NRPN, CPT, Advanced Yield Trial (AYT), Nebraska Interstate Nursery (NIN), TSWWSN, and Preliminary Yield Trials (PYT) was transplanted in May 2007 and evaluated in July Four lines with promising FHB (Form FPR07) 2

59 PI: Blodgett, Sue USDA-ARS Agreement #: resistance were included in the 2008 CPT and 10 in the 2008 AYT. We included 15 experimental lines and checks in the 2007 Tri-state FHB Nursery (South Dakota, Nebraska, and Kansas). Based on preliminary FHB disease index% average from the Kansas and Nebraska sites, our lines ranked as SD07359, SD , SD05133, SD05048, Darrell, SD02480, SD05267, SD05250, SD07288, SD05156, SD05W012, SD01058, SD07338, numbers 1, 2, 4, 5, 7, 9, 11, 13, 15, 21, 24, 32, and 36 respectively. About 2,800 head-rows and 43 EYT entries with tagged FHB QTL sources were planted in season. Best lines out of the head-row nursery will be included in the EYT in Resistant lines will be entered into regional nurseries to facilitate development of varieties with broad adaptation to the northern Great Plains. Impact: Scab is a potentially very devastating disease in the eastern and central South Dakota. Yield losses as high as 90% occurred in some fields in this area. Losses were less severe in fields planted to Expedition wheat which reflects the progress that we have made. Darrell hard red winter wheat was released in This line is expected to play a big role in eastern South Dakota where scab epidemics can be very devastating. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Producers in South Dakota now have access to superior cultivars with very good scab resistance that was not available in the past. Our effort will continue to meet the needs of our clientele. (Form FPR07) 3

60 PI: Blodgett, Sue USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Bockus, W. W., Baenziger, P. S., and Ibrahim, A. M. H Reaction of Kansas, Nebraska, and South Dakota winter wheat accessions to Fusarium head blight (FHB), Plant Disease Management Reports (online). Report 1:CF009. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. 2. Ibrahim, A.M.H., S.D. Haley, P. S. Baenziger, Y. Jin, M.A.C. Langham, J. Rickertsen, S. Kalsbeck, R. Little, J. Ingemansen, O.K. Chung, B.W. Seabourn, G.H. Bai, Ming-Shun Chen and D.V. McVey Registration of Alice wheat. Journal of Plant Registrations 2: Ibrahim, A.M.H., S.D. Haley, P. S. Baenziger, Y. Jin, M.A.C. Langham, J. Rickertsen, S. Kalsbeck, R. Little, J. Ingemansen, O.K. Chung, B.W. Seabourn, G.H. Bai, Ming-Shun Chen and D.V. McVey Registration of Darrell wheat. Journal of Plant Registrations 2: Malla, S. and A.M.H. Ibrahim Evaluation of South Dakota Winter Wheat for Fusarium Head Blight Resistance. p In Proc. South Dakota Academy of Science, Brookings, South Dakota, USA. 5. Malla, S., A.M.H. Ibrahim, and K. Glover Diallel Analysis of Fusarium Head Blight Resistance in Wheat. Journal of Crop Improvement (Accepted). 6. Malla, S., A.M.H. Ibrahim and K. Glover Winter and Spring Wheat Parental Diallel Analysis for Scab Resistance. p In Proc National Fusarium Head Blight, Kansas City, Kansas, USA. (Form FPR07) 4

61 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: William Bockus Institution: Kansas State University Address: Department of Plant Pathology 4733 Throckmorton Hall Manhattan, KS USA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Development of Scab Resistant Wheat Cultivars for Kansas. Title: FY07 ARS Award Amount: $ 26,341 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount VDUN Development of Scab Resistant Wheat Cultivars for Kansas. $26,341 Total Award Amount $ 26,341 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

62 PI: Bockus, William USDA-ARS Agreement #: Project 1: Development of Scab Resistant Wheat Cultivars for Kansas. 1. What major problem or issue is being resolved and how are you resolving it? Serious scab (Fusarium head blight) epidemics occurred in Kansas in 1982, 1990, 1993, 1995, and 2008 and annual losses average $2.1 million. Approximately one million acres of wheat in the eastern part of Kansas are annually at risk from scab where rainfall is higher during heading and corn residue is more prevalent. Since 1980, wheat acreage in the eastern one quarter of Kansas has declined by two thirds and a major cause of the decline has been farmer aversion to the risk of scab epidemics. Therefore, the availability of cultivars with resistance to scab is highly desirable for eastern Kansas. Additionally, there are three main reasons that scab has the potential to become much more prevalent in central Kansas where wheat is the traditional dominant crop (about 5 million additional wheat acres). First, there is a trend for increasing cultivation of corn in that part of the state, the main reservoir of inoculum of the scab pathogen. Second, there is a strong trend for decreasing tillage, which allows spore production and release from the residue. Third, the prevalent varieties in central Kansas are susceptible to scab. Resistant wheat cultivars adapted to this area of Kansas would also be highly desirable. Genetic resistance offers the best hope for economic management of this disease. The long-term goal of this research is to develop hard red and hard white winter wheat cultivars adapted for Kansas with improved resistance to scab. Short term objectives are to: 1) test existing local cultivars for resistance, 2) test advanced breeding lines for resistance, 3) test exotic germplasm lines for resistance, 4) test the Northern Uniform Winter Wheat Scab Nursery and the Hard Winter Wheat (Kansas, Nebraska, South Dakota) Scab Nurseries for reaction to scab, and 5) incorporate new sources of scab resistance into the Kansas wheat breeding program. Testing will be done in misted field nurseries using soil-applied infested corn grain inoculum and in the greenhouse using single-floret inoculations. Visual disease evaluation methods will be used to rate the percentage spikelets infected by the pathogen. Data will be disseminated to wheat producers and used by wheat breeders as they make selections for future Kansas cultivars. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Until involvement in the USDA Scab Initiative, there was virtually no effort to identify sources of scab resistance in Kansas breeding programs. The Initiative has resulted in the development of accurate and efficient greenhouse and field testing nurseries that are providing useful ratings for current cultivars in Kansas, advanced breeding lines, and allow participation in the regional scab nurseries. Respectively, these nurseries allow dissemination of information to growers on the reaction of current commercial cultivars, selection by breeders for scab resistance in their breeding lines, and identification of additional sources of resistance from other breeding efforts in the region that can be incorporated into Kansas breeding lines. Kansas has also taken the lead in organizing a Hard Winter Wheat Scab Screening Nursery for the hard red winter wheat breeding programs of Kansas, Nebraska, and South Dakota. This latter nursery provides valuable data on the reaction of hard red winter wheat cultivars to scab in their area of adaptation. The long-term goal of the research is to develop, deploy, and advertise winter wheat cultivars adapted for Kansas with improved levels of resistance to scab. (Form FPR07) 2

63 PI: Bockus, William USDA-ARS Agreement #: Impact: Two commercial cultivars in Kansas (Hondo and Heyne) were identified in 2000 (and confirmed in later years) as having good levels of scab resistance (3 and 4 on the 1-9 scale where 1=immune and 9=highly susceptible). During the past few years, these cultivars have averaged only 12 and 15% scab, respectively compared with about 50% in susceptible cultivars. Similarly, the cultivar Lakin has shown moderate levels of resistance with 22-34% scab. Six other commercial cultivars have also displayed moderate levels of resistance equal to, or better than, Lakin. Therefore, we have identified a few genes for scab resistance already present in cultivars adapted to Kansas that can be used by producers and may be potential sources of resistance for the development of future cultivars. Finally, both KSU wheat breeders and the USDA wheat geneticist have been involved in the project by having their breeding lines evaluated for resistance to scab. Several breeding populations are tested each year from which the breeders make selections of promising lines showing resistance. Also, there are approximately 40 advanced breeding lines (The Kansas Intrastate Nursery) that are tested each year. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Because of the scab testing efforts, a new column for reaction to Head Scab was added to the popular KSU extension publication Wheat Variety Disease and Insect Ratings for the fall, 2000 issue and has been updated in each subsequent year. For the first time, this has allowed producers in Kansas to use the reaction to scab to help select cultivars for planting. Similarly, data produced from nurseries funded by the Scab Initiative have recently been incorporated into another popular extension publication (Kansas Performance Tests with Winter Wheat Varieties). Both publications are available as hard copy or online. The involvement of breeders and the wheat geneticist has resulted in significant progress to improve the level of resistance to scab in future commercial wheat cultivars. It normally takes years to produce a new cultivar from the time initial crosses are made. The goal is to release within 4-6 years the first Kansas scabresistant cultivar produced directly from the activity of the Initiative. Additionally, this research has resulted in a germplasm release in 2004 from Kansas State University with resistance to scab derived from Triticum timopheevii ssp. armeniacum. (Form FPR07) 3

64 PI: Bockus, William USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Bockus, W. W., Fritz, A. K., and Martin, T. J Reaction of the 2006 Kansas Intrastate Nursery to barley yellow dwarf and Fusarium head blight. Plant Disease Management Reports (online). Report 1:CF012. DOI: /PDMR01. The American Phytopathological Society, St. Paul, MN. 2. Bockus, W. W., Baenziger, P. S., and Ibrahim, A Reaction of Kansas, Nebraska, and South Dakota winter wheat accessions to Fusarium head blight (FHB), Plant Disease Management Reports (online). Report 1:CF001. DOI: /PDMR01. The American Phytopathological Society, St. Paul, MN. 3. Bockus, W. W., and Davis, M. A Reaction of selected winter wheat accessions to Fusarium head blight, Plant Disease Management Reports (online). Report 1:CF013. DOI: /PDMR01. The American Phytopathological Society, St. Paul, MN. 4. Lingenfelser, J., Roozeboom, K, Fritz, A., DeWolf, E., Knapp, M., Whitworth, J., Claassen, M., Gordon, W. B., Heer, W., Kimball, J., Maddux, L., Evans, P., Long, J., Martin, T. J., Spangler, M., Schlegel, A., Chen, R., McLaughlin, L., Miller, R., Bockus, B., Shoryer, J Performance Tests. Pages 5-26 in: 2007 Kansas Wheat Seed Book. Kansas State University Agricultural Experiment Station and Cooperative Extension Service Report of Progress No pp. 5. Bockus, W. W Determining and reporting the reaction of Kansas commercial wheat varieties to wheat diseases. Poster at the meeting of the Kansas Wheat Conference, Wichita. 6. Bockus, W. W Kansas. Report for the Western Education/Extension Research Activities Coordinating Committee on cereal diseases (WERA-97). ( 7. Bockus, W. W., Fritz, A. K., and Martin, T. J Reaction of the 2007 Kansas Intrastate Nursery to barley yellow dwarf and Fusarium head blight. Plant Disease Management Reports (online). Report 1:CF006. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. 8. Bockus, W. W., Baenziger, P. S., and Ibrahim, A. M. H Reaction of Kansas, Nebraska, and South Dakota winter wheat accessions to Fusarium head blight (FHB), Plant Disease Management Reports (online). Report 1:CF009. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. 9. Bockus, W. W., and Davis, M. A Reaction of selected winter wheat accessions to Fusarium head blight, Plant Disease Management Reports (online). Report 1:CF007. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. 10. Bockus, W. W., and Wegulo, S. N Effect of foliar fungicides on Fusarium head blight of winter wheat, Plant Disease Management Reports (online). Report 1:CF011. DOI: /PDMR02. The American Phytopathological Society, St. Paul, MN. 11. Appel, J. A., De Wolf, E., Bockus, W. W., and Bowden, R. L Prelimilary 2007 Kansas Wheat Disease Loss Estimates. Kansas Cooperative Plant Disease Survey Report Bockus, W. W. and De Wolf, E. D Kansas. Report for the Western (Form FPR07) 4

65 PI: Bockus, William USDA-ARS Agreement #: Education/Extension Research Activities Coordinating Committee on cereal diseases (WERA-97). ( 13. Bockus, W. W Determining and reporting the reaction of Kansas commercial wheat varieties to wheat diseases. Poster at the meeting of the Kansas Wheat Conference, Hays. (Form FPR07) 5

66 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Robert Bowden Institution: USDA-ARS Address: Plant Science and Entomology Research Unit (PSERU) 4008Throckmorton Hall Manhattan, KS USA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Test of a Novel Control Method against Fusarium Head Blight. Title: FY07 ARS Award Amount: $ 45,000 USWBSI Individual Project(s) USWBSI Research Area * PGG ARS Adjusted Award Project Title Amount Test of Sex Pheromones as a Novel Control Method against $45,000 Fusarium Head Blight. Total Award Amount $ 45,000 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

67 PI: Bowden, Robert USDA-ARS Agreement #: NA Project 1: Test of Sex Pheromones as a Novel Control Method against Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? The germination of conidia and ascospores of Fusarium graminearum (Gibberella zeae) is inhibited by fungal sex pheromones under laboratory conditions. This suggests that pheromone peptides might be able to prevent germination and penetration of plants by F. graminearum under natural conditions. For example, a transgenic plant expressing pheromone in the florets could exhibit a novel form of Type I resistance to initial infection. There are many hurdles to overcome, but the first priority is to test the feasibility of this strategy. This proposal aims to determine optimum conditions for spore inhibition by the pheromone and then conduct infection inhibition tests by applying pheromones onto wheat spikes. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We found that germination of conidia in the laboratory was inhibited for 24 hours at concentrations of 40 μmol of the ppg1 pheromone peptide from G. zeae. The ppg1 pheromone peptide from Neurospora crassa was more effective and inhibited germination for 24 hr at a concentration of 10 μmol. Synthetic pheromone peptides were produced by substitution of amino acids from the pheromone peptide from Neurospora into the peptide from G. zeae. Some of the synthetic recombinant peptides were more effective than the natural peptides and were able to inhibit germination for 24 hr at 1 μmol. In growth chamber pathogenicity tests, 100 μmol of G. zeae pheromone peptide co-inoculated with conidia into single florets was ineffective at preventing infection. Impact: As a result of this work, we know that sex pheromones have important functions in the sexual cycle of G. zeae, the fungus that causes Fusarium head blight. Therefore, sex pheromones are a potential target for disease control. Initial attempts to inhibit conidial germination and prevent disease infection with sex pheromones were not successful. Further work is needed to define conditions affecting pheromone inhibition of germination of conidia. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The functions of sex pheromones in G. zeae have been determined. A variety of natural and synthetic fungal sex pheromone peptides have been shown to inhibit germination of conidia and ascospores in the laboratory. (Form FPR07) 2

68 PI: Bowden, Robert USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. REFEREED JOURNALS 1. Lee, J., J. E. Jurgenson, J. F. Leslie & R. L. Bowden Alignment of genetic and physical maps of Gibberella zeae. Appl. Environ. Microbiol. 74: Lee, J., J. F. Leslie & R. L. Bowden Expression and function of sex pheromones and receptors in the homothallic ascomycete Gibberella zeae. Eukaryot. Cell 7: ABSTRACTS 1. Lee, J., R. L. Bowden & J. F. Leslie Pheromone functions in Gibberella zeae. Fungal Genetics Newsletter 54(Suppl.): Lee, J., J. F. Leslie & R. L. Bowden Functions of the sex pheromones of Gibberella zeae. Proceedings of the 2007 National Fusarium Head Blight Forum (Kansas City, Missouri):30. (Form FPR07) 3

69 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Gina Brown-Guedira Institution: USDA-ARS Address: Eastern Regional Small Grains Genotyping Lab NCSU-Crop Science Campus Box 7620 Raleigh, NC USA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 25,000 Developing FHB Resistant Soft Wheat Varities by Accelerated Backcrossing. USWBSI Individual Project(s) USWBSI Research Area * HGR ARS Adjusted Award Project Title Amount Developing FHB Resistant Soft Wheat Varities by Accelerated $25,000 Backcrossing. Total Award Amount $ 25,000 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

70 PI: Brown-Guedira, Gina USDA-ARS Agreement #: NA Project 1: Developing FHB Resistant Soft Wheat Varieties by Accelerated Backcrossing. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) is a devastating disease of wheat that causes reduction in grain yield and quality in the eastern United States where soft wheat is grown. Host resistance is the best method to control losses. Although moderate FHB resistance has been found in soft red winter wheat germplasm in the eastern US, the high levels of resistance needed against this pathogen are not currently available in these cultivars. We are rapidly addressing the needs of the eastern US soft wheat growers using technology at the USDA-ARS Genotyping Lab at Raleigh, NC to combine native resistance in soft red winter wheat cultivars and breeding lines with resistance genes from Chinese sources that are tagged with DNA markers. We are attempting to complement the moderate resistance of soft wheat lines with these exotic resistance genes in an accelerated backcrossing scheme using high-throughput genotyping that should provide timely release of backcross-derived varieties in the eastern wheat region and rapidly supply improved parents for forward breeding. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We are complimenting the level of resistance to FHB that is present in soft red winter wheat cultivars in the eastern US using genes derived from Chinese wheat lines having high levels of resistance. More than 5000 backcross progeny from crosses between the soft red winter wheat (SRWW) cultivars McCormick and NC-Neuse with the highly FHB resistant lines Ning7840 and VA01W476 (Roane/W14) have been evaluated with molecular markers linked to FHB resistance genes. Genomic DNA of backcross plants having up to three resistance genes from the exotic sources was evaluated with markers distributed throughout the genome. Progeny from the backcross plants with 95-99% of the background of the adapted soft wheat parent were identified that are homozygous for different combinations of resistance genes on chromosome 3BS, 5AS and 2DL. These plants were grown in the field during the growing season to select lines with plant type similar to the SRWW parent. Lines were also advanced in the greenhouse for inbreeding and seed increase. Seeds of more than 100 F 2 - and F 3 -derived backcross lines enriched for the background of the SRWW parent that carry one to three resistance genes from the Chinese sources are being distributed to breeding programs during the summer of 2008 for fall planting. Seeds are available for replicated evaluation of these near-isogenic lines in scab inoculated nurseries that will provide important information about the effect of these resistance genes in moderately resistant soft wheat backgrounds. The backcross lines also have potential for release as cultivars. Plants with 2 to 3 pyramided resistance genes and high levels of recurrent parent background were used as parents in forward breeding by crossing to elite breeding lines adapted to the region. Marker-assisted selection was done on these three-way crosses and progeny were selected that are homozygous for FHB resistance and have genes for resistance to other pests that were segregating in the populations. Seed of the F 2:3 lines from these crosses is being distributed to collaborating breeding programs during summer These populations are a source of new lines having high levels of FHB resistance in combination with resistance to other diseases including stem, leaf and stripe rust and powdery mildew. (Form FPR07) 2

71 PI: Brown-Guedira, Gina USDA-ARS Agreement #: NA Impact: This project brings together the resources of multiple wheat breeding programs and the Eastern USDA Genotyping Lab to rapidly address the need of soft wheat growers in the eastern US. The application of high-throughput marker screening for accelerated backcrossing is allowing us to deploy exotic FHB resistance genes in adapted soft red winter wheat varieties. Seed of BC 2 F 3:4 lines similar to the recurrent parent and homozygous for FHB resistance gene combinations is being distributed to cooperating breeding programs in the region during summer Evaluation of FHB reaction of the lines that are nearisogenic for resistance gene combinations will be done during the growing season. This will provide valuable information about the effect of these resistance genes in moderately resistant soft wheat backgrounds. Field selection of lines that have potential for release as cultivars has already begun. In addition, the backcross plants selected with markers are being used as parents in forward breeding strategies to develop improved varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The germplasm being developed represents not only potential new FHB resistant varieties, but also improved parents for forward breeding and near-isogenic lines for genetic studies. The distribution of seed for evaluation and crossing is an important technology transfer event since breeders now have access to a pyramid of exotic resistance genes in an adapted soft winter wheat background. (Form FPR07) 3

72 PI: Brown-Guedira, Gina USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Costa, JM, L Al-Tukhaim, R Brown, N Gal-Edd, A Ku, E Wnger, D Van Sanford, and GL Brown-Guedira Development of scab resistant soft red winter wheat germplasm using marker-assisted selection. Proceedings of the 2007 National Fusarium Head Blight Forum, pp 175. (Form FPR07) 4

73 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Xiwen Cai Institution: North Dakota State University Address: Department of Plant Sciences Box Loftsgard Fargo, ND USA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 70,509 Characterization of Resistance to Fusarium Head Blight in Wheat and Its Relatives. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGR Enhancing Resistance to Fusarium Head Blight in Wheat Using Alien Species. $38,314 HGG Fine Mapping of Qfhs.ndsu-3AS in Durum Wheat. $ 32,195 Total Award Amount $ 70,509 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

74 PI: Cai, Xiwen USDA-ARS Agreement #: Project 1: Enhancing Resistance to Fusarium Head Blight in Wheat Using Alien Species. 1. What major problem or issue is being resolved and how are you resolving it? The lack of effective resistance to FHB and the complex inheritance of currently identified sources of resistance limit the progress of breeding for FHB resistance in wheat. We have developed and identified wheat-alien species derivatives with FHB resistance and have been introgressing resistance genes from the derivatives into adapted wheat backgrounds through chromosome manipulation. Alien introgression for FHB resistance is a time-consuming research project due to the quantitative nature of resistance and the genomic distinction between wheat and alien species. To ensure success in the alien introgression, we have been increasing the generation of the progeny derived from the crosses, manipulating wheat and alien chromosomes in the progeny, and extensively evaluating the materials of interest in each generation for FHB resistance in the greenhouse and fields. We selected 422 lines (F 5-7 ) that exhibited a FHB severity of less than or equal to 15% and favorable agronomic characteristics for seed increase in the greenhouse Summer A total of 195 advanced lines were re-evaluated for FHB resistance with three replications in the greenhouse Spring Meanwhile, these introgression lines have been grown at three locations to verify their resistance and to evaluate their agronomic performance under different field conditions. We grew 281 lines in the FHB nursery at Jianyang, China Fall Most of the lines showed similar levels of resistance as what we observed in the greenhouse screening experiments. Currently, 154 lines have been grown with three replications in the FHB nursery with a mist system in Langdon, ND. We will evaluate their reaction to FHB under that environment. Also, we have grown 285 advanced lines for seed increase and further evaluation of agronomic traits in Prosper, ND. Moreover, we haplotyped most of the introgression lines at 7 molecular marker loci linked to several FHB resistance QTL, including fhb1 on 3BS. It appears that some of the lines may contain novel resistance genes according to their haplotypes. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We developed and identified over 200 advanced alien introgression lines (BC 2 F 6-9 ) with FHB resistance under the greenhouse environments. Most of these lines consistently exhibited resistance under a field condition (Jiangyang, China). Importantly, many of the lines showed favorable agronomic characteristics, including plant height, maturity, resistance to other diseases, and yield potential, etc., in addition to FHB resistance. An obvious linkage drag has not been observed in these lines. We have been increasing seed of these lines in the field and anticipate releasing several spring wheat germplasm with FHB resistance next year. Impact: The alien introgression lines with FHB resistance and other desirable agronomic traits represent a useful source of resistance for the development of superior wheat cultivars. Utilization of these germplasm in wheat breeding will broaden the genetic basis of FHB resistance and enhance resistance of wheat cultivars to FHB. Deployment of the alien resistance genes in wheat cultivars will reduce the economic losses caused by this devastating disease. (Form FPR07) 2

75 PI: Cai, Xiwen USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Fusarium head blight resistance has been identified from relatives of wheat. It is, however, difficult to utilize the resistance genes from the relatives directly in wheat breeding. A prebreeding effort like what we have been doing in this project is needed to make the alien resistance genes available to wheat breeders. Thus, this research project has been building a bridge for wheat breeders to exploit the alien gene pool for enhancing resistance of wheat to FHB. (Form FPR07) 3

76 PI: Cai, Xiwen USDA-ARS Agreement #: Project 2: Fine Mapping of Qfhs.ndsu-3AS in Durum Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Qfhs.ndsu-3AS is a major FHB resistance QTL identified in tetraploid wheat. The map resolution of this QTL was not high enough to allow one to effectively access this locus in terms of molecular cloning and marker-assisted selection of the QTL. We have saturated the genomic region harboring Qfhs.ndsu-3AS with newly developed EST- and BAC end-derived STS and SSR markers. Also, we have generated CAPS (cleaved amplified polymorphic sequences) markers for saturation mapping of this genomic region. By collaborating with the wheat physical mapping and sequencing of 3AS project led by Dr. B. S. Gill (Kansas State University), we have identified contigs and BACs spanning the genomic region containing Qfhs.ndsu-3AS. Also, we have identified rice and Brachypodium distachyon genomic regions collinear with the QTL region on 3AS. All these efforts have significantly facilitated saturation and fine mapping of this QTL. To date, a total of 28 new molecular marker loci have been detected near the QTL. Meanwhile, we developed an F 2:3 population with 1,207 F 2 individuals/f 3 families from the cross of a resistant recombinant containing the smallest T. diccocoides chromosomal fragment containg Qfhs.ndsu-3AS in LDN background with LDN. We have been continuously developing molecular markers residing within the QTL region and using the large population to increase the resolution of the QTL map. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We detected 28 new EST- and BAC end-derived STS and SSR marker loci on 3AS. Eight of them reside near Xgwm2, a SSR marker locus tightly linked to the highest point of the QTL peak. We identified contigs and BACs harboring the marker loci in the QTL region, including Xgwm2, Xwgc501, Xwgc774, Xwgc716, and Xwgc1127, and have been assembling the contigs within this region according to the genetic map. We identified the rice and Brachypodium genomic regions collinear with the QTL region on 3AS. Also we developed a large population to further increase the resolution of the QTL map. Impact: The newly developed PCR-based molecular markers residing within the QTL region are useful for marker-assisted selection (MAS) of Qfhs.ndsu-3AS in wheat breeding and germplasm development. Some of the markers have been provided to wheat breeding programs for MAS and gene pyramiding of FHB resistance QTL. The genetic map we have constructed has been used to assemble the contigs on 3AS. These physical mapping results and the microcollinearity identified with rice and Brachypodium genomes enhance knowledge of this genomic region harboring the QTL and facilitate cloning and understanding of this QTL in durum wheat. (Form FPR07) 4

77 PI: Cai, Xiwen USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We developed new user-friendly molecular markers tagging the FHB resistance QTL Qfhs.ndsu-3AS and more precisely positioned the QTL on 3AS. This will make MAS and gene pyramiding more efficient in breeding for FHB resistance. In addition, this is the research project to first time identify the microcollinearity between the chromosomal interval containing Qfhs.ndsu-3AS and the genomic regions of rice and Brachypodium. Moreover, all the molecular markers we developed have been made available to the USDA-NRI funded project on the physical mapping and sequencing of 3AS. (Form FPR07) 5

78 PI: Cai, Xiwen USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Refereed Journal Articles: Oliver, R.E., X. Cai, R.W. Stack, T. Friesen, S. Halley, and S.S. Xu Fusarium head blight resistance in tetraploid wheat (Triticum turgidum L.). Crop Sci 48: Chen, X., J.D., Faris, J. Hu, R.W. Stack, T. Adhikari, M.E. Elias, S.F. Kianian, and X. Cai Saturation and comparative mapping of a major Fusarium head blight resistance QTL in tetraploid wheat. Molecular Breeding 19: Oliver, R.E., R.W. Stack, J.D. Miller, and X. Cai Reaction of wild emmer wheat accessions to Fusarium head blight. Crop Sci 47: Proceedings: Cai, X., S.S. Xu, R.E. Oliver, Q. Zhang, R.W. Stack,, S. Zhong, T.L. Friesen, S. Halley, and E.M. Elias Alien Introgression for FHB Resistance in Wheat - Challenges and Strategies. Proc. 11 th Intern. Wheat Genet. Symp., Brisbane, Australia, August 24-29, Xu, S.S., R.E. Oliver, X. Cai, T.L. Friesen, S. Halley, and E.M. Elias Searching for new sources of FHB resistance in the relatives of wheat. In Proc National Fusarium Head Blight Forum (abstr.), Kansas City, MO, December 2-4, Invited Oral Presentations: Zhu, X., X. Chen, J.D., Faris, J. Hu, R.W. Stack, T. Adhikari, M.E. Elias, S.F. Kianian, and X. Cai Toward a better understanding of a major FHB resistance QTL in tetraploid wheat (abstr.). Proc. National Wheat Genomics Conference, November 30 December 2, Kansas City, MO. Cai, X Molecular characterization of pest resistance and meiotic restitution in wheat. Wheat Cytogenetics Institute, Nanjing Agricultural University, Nanjing, P. R. China, April 22-23, Cai, X Wheat genetics project: identification, characterization, and introgression of desirable genes. Annual Meeting of ND SBARE and ND Wheat Commission, April 4, (Form FPR07) 6

79 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Flavio Capettini Institution: CIMMYT Address: C/O ICARDA Biodiversity & Integrated Gene Management Program P.O. Box 5466 Aleppo, Syria Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: F092 USDA-ARS Agreement ICARDA/CIMMYT FHB Barley Enhancement. Title: FY07 ARS Award Amount: $ 27,805 USWBSI Individual Project(s) USWBSI Research Area * HGR ARS Adjusted Award Project Title Amount International Barley Germplasm and Information Exchange through $27,805 ICARDA/CIMMYT. Total Award Amount $ 27,805 July 7, 2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

80 PI: Capettini, Flavio USDA-ARS Agreement #: F092 Project 1: International Barley Germplasm and Information Exchange through ICARDA/CIMMYT. 1. What major problem or issue is being resolved and how are you resolving it? The primary problems that we are working to resolve are the need for search and identification of new sources of FHB resistance in barley which will diversify the current resistance gene pool (with emphasis in 6-row types), and the need for facilitation of distribution of such resistant germplasm identified. We are meeting these needs through the following approaches: Screening new FHB resistant barley germplasm through extensive systematic screening activities of the barley genetic resources available at the ICARDA gene bank and making that available to the programs cooperating with the USWBSI. Introducing ( highly ) resistant barley germplasm from international programs and promoting germplasm exchanges, especially 6-row types, through the ICARDA gene bank and ICARDA & CIMMYT international network that otherwise maybe inaccessible to US researchers. Providing agronomically suitable FHB resistant barley germplasm to US collaborators through pre-breeding activities using major USA cultivars. Testing USA barley germplasm at CIMMYT-El Batán field station and/or through the ICARDA International Barley Improvement Network. Testing preliminary resistant gemplasm identified through other projects searching for novel sources of resistance in order to determine the GxE interaction of such sources. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: During the summer a wide screening nursery was established at El Batán, México, with artificial misting and inoculation. Disease levels reached satisfactory severity levels allowing selection of resistant genotypes. The major accomplishment was the identification of new putative sources of FHB resistance from materials that were screened, especially entries fom the ICARDA Gene Bank that were never tested before (Table 1). Genotypes tested the year before has been advanced for further testing to confirm resistance. Entries from the program selected in 2006 were distributed to US breeding programs (North Dakota State University, The University of Minnesota, Busch Agricultural Resources, Inc.) and Canada (Agriculture Canada at Brandon, Manitoba). Three nurseries that were deployed were the EGS2007: 282 entries, NABSEN 2007: 8 entries contributed to the nursery, and China Nursery 2007: 100 entries. Genotypes with at least 2 seasons of selection were the ones tested in the China Nursery. Germplasm with superior resistance is being used in crosses within the breeding program. Genotypes showing unusual brightness were identified in China within the entries of BARI and ICARDA. Seed samples of those exceptional entries were distributed by BARI to interested breeders during the Barley CP meeting in Minnesota. (Form FPR07) 2

81 PI: Capettini, Flavio USDA-ARS Agreement #: F092 Table 1. Number of nurseries and entries screened at El Batán, México during 2007 and number of putative resistant entries selected for further testing. Name Origin Entries Selected MV-06 New Germplasm ICARDA 2006 ICARDA Gene Bank Bulgaria Bulgaria 5 3 Germplasm Introduced from Other Programs Alberta 2007 CANADA BMZY-07 F BARI 2007 BARI Brandon 2007 Canada BARI 2006 USA NABSEN 07 USA Germplasm from the Breeding Program Preliminar FHB 07 Breeding Program EGS 2007 Breeding Program China 2007 Breeding Program F10CEBADASCX Breeding Program 8 7 Alberta Best Canada 7 6 Palestina ICARDA Gene Bank 3 1 ICARDA 2006 ICARDA Gene Bank (Sel.) Eritrea/Ethiopia ICARDA Recolection BARI Sel Obregón Total Impact: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community is basically obtaining: 1. Putative resistance sources from ICARDA gene bank that was not available before. 2. Advanced lines originated from the ICARDA/CIMMYT breeding program with enhanced FHB resistance as well as resistance to several other important diseases in an acceptable agronomic background, many of them in a US-germplasm based lines. (Form FPR07) 3

82 PI: Capettini, Flavio USDA-ARS Agreement #: F092 Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Capettini Flavio, Stefania Grando, Les Wright, Linnea Skoglund Contribution of ICARDA to Improve Fusarium Head Blight Tolerance in Barley 20 Years of International Collaboration. The 10th International Barley Genetics Symposium, Alexandria, Egypt, 5-10 April Capettini, Flavio, Mario Cattáneo, Silvia Germán, Luz Gómez Pando, Euclydes Minella, Sergio Pieroni, Miguel Rivadeneira, Mauro Zamora Barley enhancement in Latin America in the last 20 years: a story of success. In Promaco PTY LTD. 13 th Australian Barley Techincal Symposium. Fremantle, Western Australia August pp (Form FPR07) 4

83 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Jose Costa Institution: University of Maryland Address: Dept. of Nat. Res. and Landscape Arch Plant Sciences Building College Park, MD Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 27,661 Screening and Developing Wheat Germplasm with Resistance to Scab. USWBSI Individual Project(s) USWBSI Research Area * HGG VDUN ARS Adjusted Award Project Title Amount Sources of Variation for DON Concentration in Moderately Resistant SRWW. $4,917 Developing and Screening Wheat Germplasm with Resistance to Scab. $ 22,744 Total Award Amount $ 27,661 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

84 PI: Costa, Jose USDA-ARS Agreement #: Project 1: Sources of Variation for DON Concentration in Moderately Resistant SRWW. 1. What major problem or issue is being resolved and how are you resolving it? In collaboration with Dr. Caly Sneller (Ohio State University) we are attempting to understand variation for DON accumulation among moderately resistant lines to efficiently obtain cultivars with low scab index and low DON required by the wheat industry. We are testing a large set of soft red winter wheat breeding lines with moderate resistance to FHB. During the we grew these lines, their parents, and checks, and rated them for FHB in June 2007 at Feekes GS We collected grain samples that will be tested for DON in the fall of List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): At this time we are processing grain samples harvested in June 2008 for DON testing. No major accomplishment has been achieved at this time. It is the first season of testing in MD for this project. (Form FPR07) 2

85 PI: Costa, Jose USDA-ARS Agreement #: Project 2: Developing and Screening Wheat Germplasm with Resistance to Scab. 1. What major problem or issue is being resolved and how are you resolving it? The major problem being addressed is the need to develop rapidly and effectively host resistance to scab (Fusarium Head Blight) from exotic sources into adapted soft red winter wheat (SRWW) germplasm. The approach to address this problem is to backcross, and to three-way cross the Sumai 3 allele and other exotic resistance alleles into adapted SRWW lines and varieties. Marker-assisted selection (MAS) is being used to rapidly incorporate Sumai 3 (from Ning7840) resistance into SRWW lines such as McCormick, that have wide adaptation in the Southern and Eastern US wheat growing regions and moderate resistance to scab. Over 400 Backcross 2 F1 seedlings were screened for SSR markers at the USDA in Raleigh (NC) in collaboration with Dr. Gina Brown-Guedira at that National Genotyping Center. Of these, a single BC2F1 plant was selected that had Ning7840 alleles at the 3BS, 5A and 2DL genomic regions. 340 F2 progenies were further screened in 2007, selected homozygous BC2F3s were planted in the field in the fall of 2007 in our inoculated nursery. Incidence and severity of scab were scored in the spring of 2008 and these will be tested for DON in the fall of Additionally, selected BC-1F1s were crossed with the wheat cultivar SS 8641 that has wide adaptation as well as leaf and stripe rust resistance. The BC1 F1 seeds were screened with markers in and the BC1-F2s selected progenies were advanced in the growth chamber. Selected homozygous BC1F3s for the 3BS, 5A and 2DL genomic regions, will be planted in the field in the fall of Furthermore, screening of MD (University of Maryland) wheat advanced lines and check varieties was conducted under field conditions in an inoculated nursery at Salisbury (MD). Conditions favorable for disease development were aided with daily misting before and during wheat flowering. The scab inoculum was scabby corn grain spread in the field a month before flowering. The Southern wheat scab and Northern Uniform Scab Screening nurseries that include new experimental lines were also screened for resistance at Salisbury (MD) with artificial inoculation and misting. Data for all nurseries was obtained for scab incidence, scab severity, Fusarium damaged kernels, seed weight, plant height, heading date, and DON levels. (Form FPR07) 3

86 PI: Costa, Jose USDA-ARS Agreement #: List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Incorporation of the 3BS, 5A and 2DL quantitative trait loci (QTL) of resistance to scab from Sumai3 into adapted soft red winter wheat germplasm. Impact: the availability of these germplasm with resistance will reduce scab negative effects in years favorable to scab development. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? Plant breeders will have homozygous F4 seed for the 3 QTL of adapted soft red winter wheat germplasm available in the McCormick and SS8641 background in the fall of 2008 for crossing. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Costa, J. M., Al-Tukhaim, L., Gal-Edd, N., Ku, A., Wenger, E., Brown-Guedira, G., and Van Sanford, D Development of Scab Resistant Soft Red Winter Wheat Germplasm (SRWW) Using Marker-Assisted Selection (MAS). Proceedings of the 2007 National Fusarium Head Blight Forum p Kansas City, MI. Presentations: Costa, J.M Use of DNA markers for developing head blight resistant wheat genotypes Argentina Symposium on wheat diseases. University of La Plata, Argentina. 11/23/2007. Costa, J.M Marker-assisted selection for head blight resistance in winter wheat. University of Puerto Rico. Mayaguez, PR. 1/17/2008. (Form FPR07) 4

87 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Christina Cowger Institution: USDA-ARS Address: Plant Science Research Unit 3409 Gardner Hall Box 7616 Raleigh, NC Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 43,000 Effects of Post-Anthesis Moisture, Cultivar, and Infection Timing on FHB and DON in Wheat. USWBSI Individual Project(s) USWBSI Research Area * EEDF ARS Adjusted Award Project Title Amount Effects of Post-Anthesis Moisture, Cultivar, and Infection Timing $43,000 on FHB and DON in Wheat. Total Award Amount $ 43,000 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

88 PI: Cowger, Christina USDA-ARS Agreement #: NA Project 1: Effects of Post-Anthesis Moisture, Cultivar, and Infection Timing on FHB and DON in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Deoxynivalenol (DON) levels are important both for their health effects and because DON is a pathogenicity factor in cereals. Our knowledge of the epidemiological and host genetic influences governing DON concentrations is incomplete. While anthesis is thought to be the primary period for FHB infection in wheat, late infections can also lead to DON production. High levels of DON have sometimes been observed in the absence of abundant disease symptoms. Visual disease severity and DON concentration in grain are often correlated, but coefficients are generally low, and vary greatly among locations and years. Our research goal is to improve our understanding of how moisture duration and infection timing affect disease development, Fusarium growth, and DON production. Our multi-year, replicated field experiment involved seven winter wheat cultivars with varying resistance levels and types in a misted nursery with 4 durations of misting and 4 inoculation timings. In addition to assaying DON at harvest time, we have monitored DON levels in spikes from flowering until about 10 days after normal harvest time. Taken together, the data will enhance our ability to forecast epidemic severity and economic risk. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The two years in which disease levels were adequate in our inoculated winter wheat field experiment were 2006 and In both years, FHB incidence, FHB severity, and mean grain DON increased significantly as duration of post-flowering misting increased up to 20 days. We also found that grain DON at harvest exceeded acceptable levels in heads infected at flowering and in heads infected 10 days after flowering, but not in those infected 20 days after flowering. Further, we observed a high DON in plump kernels scenario in heads inoculated 10 days after flowering and subjected to extended misting during grain-fill. In other words, in this treatment combination, DON exceeded acceptable levels while visually scabby kernel (VSK) percentages remained indistinguishable from the low levels found in unmisted treatments. In a time-course study, we found that DON levels declined during grain-fill up to and in some cases during the harvest period. This finding led to our current investigation of whether harvest delay results in increased, decreased, or unchanged DON levels, a question we are addressing under varying levels of harvest-time moisture. This subject is of considerable practical relevance to growers. Our findings will help increase the accuracy of FHB risk forecasting, relate visual FHB symptoms to grain DON levels at harvest, and construct models that accurately forecast the risk of unacceptable DON levels at harvest. The information will be used by USWBSI risk modelers, and by agricultural scientists and extension workers helping growers understand the factors that lead to FHB and DON damage. (Form FPR07) 2

89 PI: Cowger, Christina USDA-ARS Agreement #: NA Impact: This is the most systematic investigation ever conducted of the interaction of wheat resistance genotype, FHB infection timing, and post-flowering moisture. Capturing data on disease, kernel infection, and DON throughout the grain-fill period gives us greater insight into the process of disease and toxin development. The finding of increased kernel infection, visual symptoms, and DON with increasing days of post-flowering moisture is an important and novel contribution to our current U.S. national FHB risk forecasting program, which focuses on pre-flowering weather conditions and corn residue. The impact of this research will be a greater capacity to forecast, monitor, and manage DON in wheat. Researchers and extension workers will be better able to advise growers and millers when to anticipate and prepare for problems with DON, and when DON management measures should be prioritized. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Researchers, extension workers, and growers now have strong evidence that post-flowering moisture duration up to three weeks in length enhances disease severity and DON concentrations in wheat grain. This knowledge will help researchers develop and refine DON risk forecasting models. Our data are also consistent with the hypothesis that cultivar resistance may interact with post-flowering moisture duration, although this needs further investigation. Such models will help growers and millers reduce and manage DON risk and damage. Our results will also eventually help breeders understand whether advanced lines should be screened under extended post-flowering irrigation. (Form FPR07) 3

90 PI: Cowger, Christina USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Duration of post-flowering moisture can affect FHB in wheat Cowger, C. and Medina- Mora, C. In: Canty, S., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the National Fusarium Head Blight Forum; 2007 Dec 2-4, Kansas City, MO. Lexington: University of Kentucky. (Conference Proceedings) Effects of post-flowering moisture on Fusarium head blight and deoxynivalenol levels in winter wheat, 200_. Cowger, C., Patton-Özkurt, J., and Brown-Guedira, G. Phytopathology (in preparation). (Form FPR07) 4

91 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Erick De Wolf Institution: Kansas State University Address: Dept. of Plant Pathology 4607 Throckmorton PSC Manhattan, KS Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 34,695 Prediction Models and Improved Pre-Harvest Estimates of Deoxynivalenol. USWBSI Individual Project(s) USWBSI Research Area * EEDF ARS Adjusted Award Project Title Amount Prediction Models and Improved Pre-Harvest Estimates of $34,695 Deoxynivalenol. Total Award Amount $ 34,695 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

92 PI: De Wolf, Erick USDA-ARS Agreement #: Project 1: Prediction Models and Improved Pre-Harvest Estimates of Deoxynivalenol. 1. What major problem or issue is being resolved and how are you resolving it? Wheat producers need a reliable disease prediction system to help evaluate the risk of disease and potential need for fungicide applications. Fungicides are an important part of the integrated management for head scab, but are not needed when the disease risk is low. Making the correct decision to apply or not apply improve the potential profit for growers, and helps ensure the grain has lowest possible levels of DON. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Disease prediction models were deployed in 24 states where head scab epidemics have occurred. Accuracy of the prediction models has been stable at approximately 75 to 80% depending on the type of wheat and production region. We are now testing the first generation of a new type model that should allow us to address parts of the pathogen biology that are not currently part of the prediction system. These new models incorporate a module to predict DON for the first time. Impact: Wheat growers in 24 states have access to prediction models for head scab. Growers use these models to assess the risk of disease and respond with fungicides if needed or avoid pesticide use when the risk of disease is low. The new modeling approach should be able to improve the accuracy of disease predictions. The new models will also incorporate DON into the predictions. These enhancements will increase the value of the prediction models as decision tools used by growers. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Wheat growers in 24 states have access to prediction models the help them make disease management decisions based on real-time weather information. (Form FPR07) 2

93 PI: De Wolf, Erick USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Paul, P.A., Lipps, P.E., De Wolf, E.D., Shaner, G., Buechley, G., Adhikari, T., Ali, S., Stein, J., Osborne, L. and Madden, L.V A distributed lag analysis of the relationship between Gibberella zeae inoculum density on wheat spikes and weather variables. Phytopathology 97: De Wolf. E.D. and Isard, S.A Disease cycle approach to plant disease prediction. Annual Review of Phytopathology 45: (Form FPR07) 3

94 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Ruth Dill-Macky Institution: University of Minnesota Address: Department of Plant Pathology 495 Borlaug Hall St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 108,780 Crop Residue Management and Screening Techniques for Improved Management of FHB. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Targeting Fusarium-infested Crop Residues in the Control of Fusarium Head Blight. $34,146 EEDF Factors Influencing the Accumulation of DON in Fusarium-Infected Wheat. $ 42,439 HGR FHB Resistance in Hard Red Spring Wheat Near-Isogenic Lines. $ 32,195 Total Award Amount $ 108,780 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

95 PI: Dill-Macky, Ruth USDA-ARS Agreement #: Project 1: Targeting Fusarium-infested Crop Residues in the Control of Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? Reduced tillage practices, which have been adopted worldwide in agriculture, have undoubtedly contributed to the global upsurge in FHB of wheat and barley. This study is investigating our ability to reduce the inoculum associated with Fusarium-infested residues. Targeted are treatments that either a) promote the rate of decomposition of crop residues (particularly the residues of Bt-corn - which decomposes more slowly than those of regular corn residues), or b) reduce the survival of the Fusarium spp. which infest crop residues. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Three field experiments (examining crop residues from wheat, barley and corn, respectively) were established during the 2007 growing season in Rosemount, MN. The residues produced were used to establish treatments designed to examine the effect of biological control agents (Trichoderma spp. - fall application), fungicides (prothioconazoletebuconazole mix - spring application) and soil amendments (bentonite clay, urea, and spent lime (spent lime is a byproduct of beet processing and has been shown to reduce Aphanomyces root rot in sugar beets) - fall application). Additionally, in the corn residue trial, fine mechanical chopping of residues was introduced as an additional treatment. The residue cover of the soil surface of these trials was monitored in spring. Additionally the residue in these trials is currently being sampled to determine its inoculum potential. Results from the first sampling this spring indicated that none of the treatments significantly impacted the soil cover provided by the residues before the establishment of the subsequent wheat crop. Assessments of the treatment effects on inoculum potential are ongoing using traditional mycological techniques to examining ascospore production and PCR for the identification of the Fusarium species in the residues. As part of the PCR identification in this study, suitable protocols for extracting DNA from the residues collected on the soil surface, and subsequently PCR testing, needed to be developed. This has now been accomplished. FHB incidence within the wheat sown into each trial in the 2008 season will assessed at the appropriate to stage of crop maturity. The crops which will provide the residues for the planned 2008/2009 field experiments have been established and inoculated in preparation for Impact: Preliminary results indicate that the treatments employed here do not reduce the residues on the soil surface in the spring - this is actually desirable as residue cover is required for soil conservation programs. The effect on the treatments examined on residue decomposition the inoculum load from these residue/treatment combinations, is however still to be determined and the analysis of sampled materials is in progress. The development of practical and repeatable protocols for DNA extraction and PCR testing from residues will enable the planned work in this study to be completed and will greatly aid subsequent work in this area. (Form FPR07) 2

96 PI: Dill-Macky, Ruth USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The findings of this research have the potential to impact our understanding of FHB and may aid in the development of integrated management strategies for FHB. Specifically the study will indicate if the fine chopping of corn residues or the application of soil amendments can be used to accelerate the decomposition of crop residues or the inoculum production of these residues. The study should also indicate if soil amendments, biological control agents, or fungicides, as tested in this study, could be used to target the survival of Fusarium spp. in crop residues. (Form FPR07) 3

97 PI: Dill-Macky, Ruth USDA-ARS Agreement #: Project 2: Factors Influencing the Accumulation of DON in Fusarium-Infected Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Few studies have closely examined the development of Fusarium head blight (FHB) and deoxynivalenol (DON) accumulation in relation to the resistance of wheat cultivars, the relative aggressiveness of F. graminearum isolates, and the competence and ability of F. graminearum isolates to produce DON, or the impact of environmental conditions, especially moisture on the accumulation of DON in Fusarium-infested wheat. This project aimed to improve our knowledge of the development of Fusarium head blight and the accumulation of DON in wheat by examining; i) the effect of environmental conditions, principally moisture between anthesis and harvest, on the development of FHB and the accumulation of DON in wheat; ii) the impact of host genetic resistance on the development of FHB and the accumulation of DON in wheat; and iii) the effect of pathogen variability (aggressiveness and mycotoxin production capacity) on the development of FHB and the accumulation of DON in wheat. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: An inoculated, mist irrigated field experiment examining FHB development and DON accumulation was established in St. Paul in Entries were planted in a randomized split-split-plot design with five replicates. Main plots were irrigation regime (irrigation for 14, 21, 28 and 35 days after inoculation (dai)), subplots were host genetic background and sub-sub-plots were F. graminearum isolates. Plots were inoculated at anthesis and mistirrigated following inoculation for 14 days to facilitate disease development and then irrigation treatments were imposed. Entries were assessed for FHB incidence and severity and grain harvested and assessed for visually scabby kernels (VSK) and DON. Developing spikes were harvested at regular intervals between inoculation and maturity and assessed for FHB symptoms and DON. Impact: Severity, VSK and DON, across all isolates, were significantly higher in the susceptible wheat Wheaton. FHB severities were not significantly impacted by irrigation treatments, though this was to be expected as the first irrigation treatments was only imposed 14 days after inoculation and disease assessment was at 21 dai. VSK values were significantly lower in the treatments receiving the least amount of mist-irrigation (14 DAI) suggesting that extended moisture promotes disease development after disease assessment in the field. DON was significantly lower in the longest (35 dai) misting treatment. DON in head samples was reduced with increased durations of irrigation, but was only significantly lower in grain from the 35 dai misting treatment. The reduction of DON was larger in Wheaton than other cultivars. Our results suggest that DON may be reduced by late-season moisture despite increased grain colonization. Leaching may explain the reduction of DON observed with an increased misting duration, and differences in tissue morphology and metabolism may determine the rate of leaching from specific tissues. (Form FPR07) 4

98 PI: Dill-Macky, Ruth USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The finding of this study demonstrate that rainfall prior to harvest influences the DON levels in Fusarium-infected grain. The findings increases our understanding of why there are discrepancies between; a) visual evaluations of FHB made in the field prior to head senescence, b) post harvest visual assessment of FHB damage to grain (VSK or FDK), and c) the DON content of harvested grain. These discrepancies are frequently observed in breeder's FHB screening nurseries and in commercial fields where apparently sound grain is found to contain significant levels of Fusarium-produced mycotoxins, especially DON. Understanding the impact of different host resistance genes on DON accumulation will be utilized to establish the relationship between resistance to disease development and toxin accumulation and evaluate the value of host resistance genes. An understanding of the impact of the variability within the pathogen population for aggressiveness and DON toxin as measured under controlled environmental conditions on DON production in the field will be used to evaluate the magnitude of the threat posed by potential shifts in the pathogen population with respect to aggressiveness and/or the spectrum and quantities of mycotoxins produced. (Form FPR07) 5

99 PI: Dill-Macky, Ruth USDA-ARS Agreement #: Project 3: FHB Resistance in Hard Red Spring Wheat Near-Isogenic Lines. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) resistance quantitative trait loci (QTL) have been mapped in a diverse range of common wheat (Triticum aestivum) genotypes and related species, but most have not yet been utilized in commercial wheat. This project is developing wheat lines that are principally hard red spring wheat (HRSW), but are near-isogenic for the different reported novel QTLs (QTL NILs) from unadapted or alien germplasm. We are using the most rapid method - marker-assisted backcrossing - to introduce new FHB resistance QTLs into HRSW. Resultant materials can then be used to critically assess whether these presumptive QTLs do actually increase FHB resistance in HRSW. Further, pyramiding FHB resistance QTLs is expected to provide a greater opportunity to increase FHB resistance in HRSW. We will use the QTL NILs with different individual FHB resistance QTLs to pyramid these QTLs, and thus identify favorable QTL combinations. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Introgression of exotic Fusarium head blight (FHB) resistance QTLs into elite wheat gene pools is expensive and time consuming, and so it is beneficial to know in advance if the effort to deploy a new QTL in a breeding program is worth the effort. Greenhouse and field studies using multiple backcross near-isolines of hard red spring wheat identified individual exotic FHB resistance QTLs that appear to improve FHB resistance, as well as a set of QTLs that do not. Pyramided combinations of two QTLs that appear to be effective individually and linked to the markers use for their introgression were developed, and sets of F 3 families with 0, 1 or 2 of the QTLs were developed. These materials will be used for future studies of FHB resistance in the field. One promising new putative FHB resistance QTL is not linked to the introgressed marker, leading us to embark on a marker identification effort so that it can be used for further gene pyramiding. Impact: These near-isolines provide FHB resistance QTL validation for U.S. hard red spring wheat breeding programs, and thus are a prebreeding resource for them. They represent useful genetic stocks for a plethora of uses in the exploration of wheat-fusarium interactions. Materials have been distributed to wheat researchers in 6 U.S. states and the U.K. for a diversity of research uses. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: This is, to our knowledge, the largest available set of wheat backcross near-isolines for different reported FHB resistance QTLs. The information gained on individual QTL effects provides a breeding guide to FHB resistance QTL selection and introgression efforts. The wheat FHB research community also has access to these materials for additional experimental purposes. (Form FPR07) 6

100 PI: Dill-Macky, Ruth USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Di, R., Blechl A., Dill-Macky, R., Tortora, A. and Tumer N.E. (2007). Expression of the N- terminal 99 amino acids of yeast ribosomal protein L3 in transgenic wheat confers resistance to Fusarium head blight. Mol. Plant Breed., 5:283. Di, R., Blechl, A., Dill-Macky, R., Tortora, A. and Tumer, N. (2007). Expression of a truncated form of ribosomal protein L3 in transgenic wheat confers resistance to deoxynivalenol and Fusarium Head Blight. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p. 48. Dill-Macky, R., Dong, Y. and Schwarz, P. (2007). Sampling Grain for Deoxynivalenol (DON) Analysis: A Researchers Guide. (PDF) Posted 7/16/07 on the US Wheat and Barley Scab Initiative Website. ( Gautam, P. and Dill-Macky, R. (2007). Fusarium Head Blight infection and deoxynivalenol production by Fusarium graminearum in wheat. Phytopathology, 97:S39. Gautam, P. and Dill-Macky, R. (2007). Effect of post inoculation moisture on deoxynivalenol accumulation in Fusarium graminearum-infected wheat. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p Makandar, R., Nalam, V., Essig, J.S., Schapaugh, M.A. Trick, H., Dill-Macky, R. and Shah, J. (2007). Enhancing Fusarium Head Blight resistance in wheat by manipulating mechanisms contributing to host resistance and susceptibility. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p Schwarz, P., Dong, Y. and Dill-Macky, R. (2007). Deoxynivalenol measurement: Sources of error and sampling recommendations. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p. 13. Shin, S.H., Lewis, J.M., Mackintosh, C.A., Elakkad, A.M. Wennberg, K.J., Heinen, S.J., Dill- Macky, R. and Muehlbauer, G.J. (2007). Transgenic wheat with enhanced resistance to Fusarium Head Blight. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p. 63. Rojas-Barros, P., Blankenheim, Z.J., Wennberg, K.J., Elakkad, A.M., Dill-Macky, R. and Garvin, D.F. (2007). Fusarium head blight resistance of wheat lines near-isogenic for five different FHB resistance QTLs. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, Missouri, USA, December 2-4, 2007, p (Form FPR07) 7

101 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Yanhong Dong Institution: University of Minnesota Address: Department of Plant Pathology 495 Borlaug Hal St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Diagnostic Services for DON. Title: FY07 ARS Award Amount: $ 169,842 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount FSTU-S Diagnositc Services for DON. $169,842 Total Award Amount $ 169,842 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

102 PI: Dong, Yanhong USDA-ARS Agreement #: Project 1: Diagnositc Services for DON. 1. What major problem or issue is being resolved and how are you resolving it? Our laboratory provided deoxynivalenol (DON) and related mycotoxin diagnostic services for Fusarium Head Blight (Scab) research projects conducted by 34 PIs in 14 states last year. The major issue that we dealt with was how to efficiently handle huge amounts of samples submitted by so many groups and ensure researchers to get their results in a timely manner. In general, we analyzed samples based on a first come, first served policy. In case we received large amounts of samples from a single group or received several submissions from different groups around same time, we contacted PI(s) about their desired dates of having DON results for each set of their samples and adjusted samples analysis schedules to make sure that each PI could receive their results in a reasonable time frame. By doing so, we were able to provide DON results to most PIs within one month after receiving their samples. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: From May 2007 to April 2008, the Mycotoxin Diagnostic Laboratory at the University of Minnesota analyzed 29,217 samples, which was 2.1 times the number of samples analyzed last year (13,873) and 42.5% more than the estimate (20,500) presented in the proposal. The huge increase of samples reflected the great needs of DON analysis by the scab research community. The samples were submitted by 34 scab research groups from 14 states including Minnesota, Kansas, Michigan, New York, Arkansas, Indiana, South Dakota, Illinois, Kentucky, Louisiana, Ohio, North Carolina, Maryland, and Virginia. They included 20,299 regular mature grain samples (6-100 g) and 8,918 small size samples such as single kernels, single spikeletes, single heads, small stems, and fungal cultures extracts. The target toxins included DON, 15-Acetyl-DON, 3-Acetyl-DON, nivalenol and zearalenone. Ergosterol, a chemical marker for measuring fungal biomass, was also analyzed for some samples as requested by researchers. (Form FPR07) 2

103 PI: Dong, Yanhong USDA-ARS Agreement #: Impact: The DON data has been used in all areas of scab research. By analyzing mycotoxins, the project provided support to barley and wheat breeding programs to develop resistant varieties, and to researchers to study disease mechanisms and to develop effective and economical chemical and biological disease controls. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Mycotoxin data provided to scab researchers by our laboratory gave researchers a means to evaluate the effectiveness of their efforts in fighting Fusarium Head Blight. (Form FPR07) 3

104 PI: Dong, Yanhong USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Goswami, R. S.; Dong, Yanhong; Punja, Z. K. Host range and mycotoxin production by Fusarium equiseti isolates originating from gindeng fields Can. J. Plant Pathol. 2008, 30, Jiang, Guo-Liang,; Dong, Yanhong; Shi, Jinrong; Ward, Richard W. QTL analysis of resistance to Fusarium head blight in the novel wheat germplasm CJ II. Resistance to deoxynivalenol accumulation and grain yield loss Teror. Appl. Genet., 2007, 115, Yu, J.-B.; Bai, G.-H; Zhou, W.-C.; Dong, Yanhong; Kolb, F. L. Quantitative Trait Loci for Fusarium Head Blight Resistance in a Recombinant Inbred Population of Wangshuibai/Wheaton Phytopathology, 2007, 98, Kong, L.; Dong, Yanhong; Ohm, H. W. Characterization of Resistance to Deoxynivalenol (DON) Accumulation in Different Wheat Lines Proceedings of the 2007 National Fusarium Head Blight Forum, 2007, 191. (Form FPR07) 4

105 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Floyd Dowell Institution: USDA-ARS Address: 1515 College Avenue Manhattan, KS Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Single Kernel Sorting Technology for Enhancing Scab Resistance. Title: FY07 ARS Award Amount: $ 25,000 USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Single Kernel Sorting Technology for Enhancing Scab Resistance $25,000 and Grain Quality. Total Award Amount $ 25,000 Floyd Dowell 7/11/08 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

106 PI: Dowell, Floyd USDA-ARS Agreement #: NA Project 1: Single Kernel Sorting Technology for Enhancing Scab Resistance and Grain Quality. 1. What major problem or issue is being resolved and how are you resolving it? Wheat scab caused by fungi of Fusarium spp has become a major constraint in the wheat industry by affecting crop yields and quality of grain products. Development of Fusarium resistant/tolerant wheat varieties is a cost effective and safe way to address this critical issue facing the wheat industry. Objective identification of scabby, or Fusarium Damaged Kernels (FDKs) and estimation of the DON levels of FDKs are two important things needed in evaluating varieties for scab resistance or for assessing grain samples for quality. At present, visual methods are used to classify FDKs from sound kernels. However, it is possible for visually-sound kernels to have higher DON levels while visually-scabby kernels can be free from DON. Visual methods alone are not adequate to identify FDKs. Estimation of DON levels is expensive, requires numerous chemicals for extracting and assaying DON, and is also difficult to perform on single kernels. Near Infrared Spectroscopy (NIRS) can be used as a rapid and non destructive method to identify FDKs and to estimate DON levels of single kernels rapidly. Hence we are developing NIRS techniques to identify FDKs and estimate DON levels in single wheat kernels in order to facilitate evaluation of germplasm for varietal improvement. Single kernel scab and DON calibrations were developed using an automated single kernel NIR system for classification of sound and FDKs and to estimate DON levels of single wheat kernels. Attempts are now being made to minimize error levels in single kernel DON estimation. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: In sorting 108 wheat samples from NDSU using the scab calibration for sorting FDKs from sound kernels, our NIR technique sorted visibly sound kernels with a high accuracy (98.75 %). Samples were sorted into fractions with: 1) sound kernels {No detectable DON or low DON levels}; 2) FDKs with medium DON levels; and 3) FDK with high DON levels. Further detailed DON analysis of the sorted fractions of 23 scabby wheat samples from NDSU indicated that sorted sound fraction contained ppm DON (Mean= 0.8 STD=1.8) while FDK with medium DON fractions had ppm DON (Mean=98.2 STD=58.3) and the FDK fraction with high DON levels had ppm DON (Form FPR07) 2

107 PI: Dowell, Floyd USDA-ARS Agreement #: NA (Mean=680.1 STD=251.0) showing that NIR technique could separate sound from FDKs successfully and further classify FDKs into groups depending on the DON levels. Single kernel DON estimation work showed that our NIR technique can sort FDKs from sound kernels and estimate the DON levels of fractions having higher DON levels (>50 ppm) fairly successfully. However, further studies are necessary before it can be used for accurate DON estimation in kernels having low DON levels. Impact: This technology can be used to sort sound kernels from FDKs. Further FDKs could be sorted into 2-3 fractions based on the severity of scab/don levels. Hence it is possible to use this technology to evaluate germplasm for Fusarium resistance and will help breeders evaluate many lines to identify superior breeding materials. This technique can also be used to select single kernels that may have scab resistance. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Currently scabby kernels are identified manually and subjectively by trained personal based on visual symptoms. Now an objective method is available for wheat breeders to identify scab damaged kernels based on DON levels. This is a rapid, non destructive and environmentally safe method. Breeders can use the assessed kernels for generation advancement or for other trait analysis. (Form FPR07) 3

108 PI: Dowell, Floyd USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peiris, K.H.S, M.O. Pumphrey, Y. Dong and F.E. Dowell NIRS method for precise identification of Fusarium damaged wheat kernels. AACCI Poster, September 21-24, Honolulu, HI. A poster on the ongoing work in relation to scab sorting presented to the participants of Hard Winter Wheat Working Group meeting held in Manhattan, KS, May (Form FPR07) 4

109 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Alan Dyer Institution: Montana State University Address: Dept. of Plant Sciences and Plant Pathology 119 Ag Bioscience Facility Bozeman, MT Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Responding to Montana's Head Scab Epidemic. Title: FY07 ARS Award Amount: $ 20,488 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount VDUN Responding to Montana's Head Scab Epidemic. $20,488 Total Award Amount $ 20,488 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

110 PI: Dyer, Alan USDA-ARS Agreement #: Project 1: Responding to Montana's Head Scab Epidemic. 1. What major problem or issue is being resolved and how are you resolving it? There are 540,000 irrigated production acres in the northern Rocky Mountain range and western Great Plains that are potentially impacted by Fusarium Head Blight (FHB). Many growers in scab-infested areas are converting to FHB tolerant varieties from public and private companies. However, the current FHB resistant varieties are tall in stature and weak-strawed. The objective of the spring wheat breeding effort is to develop FHB-resistant varieties suitable for production in high-yield, irrigated systems. The FHB nursery is located on a producer s field with a history of FHB disease and a uniform inoculum source of F.pseudograminearum and F. graminearum from the wheat residues. In the 2006 and 2007, the overall grain deoxynivalenol (DON) concentration of the nursery was 1.88 ppm. Susceptible varieties had 0.6 to 2.0 ppm grain DON levels, including; Hank, Vida, Howard, Explorer (HWW), Choteau and Expresso. Tolerant varieties had less than 0.5 ppm grain DON, including; Kuntz, Volt, Freyr, Granite, Knudson, Alsen, Glenn, Kelby, and an experimental line MT0550 (Choteau/ND709-9). Tolerant varieties had 8% incidence of symptomatic scab heads as compared to 28% among varieties lacking the Sumai3 gene. Overall, resistant varieties yielded 80.4 bu/ac or 18% higher than varieties lacking tolerance to FHB. Grain test weights were 61.8 lb/bu in the tolerant varieties and 57.6 lb/bu in the susceptible varieties. We have used molecular markers to backcross the Sumai 3 QTL into three backgrounds of spring wheat. The Sumai 3 QTL has performed well under Montana scab disease conditions. We have used molecular markers to backcross the Sumai3 QTL into Choteau, a solid-stem variety with resistance to the wheat stem sawfly and which has performed very well in irrigated production. Molecular marker GWM 533 was utilized for selection in Choteau lines. An FHBresistant line similar to Choteau should find immediate utility in irrigated production areas as well as in sawfly infested areas of eastern Montana. Another line used as recurrent parent for backcrossing include MT0249, a variety with long green leaf duration and short stature. MT0249 is a sister line to our recent cultivar release Vida but is about 6 cm shorter under irrigation. BARC 133 was used when MT0249 was a recurrent parent. Sufficient seed of Choteau and MT0249 BC3-derived lines will be available for single hill plot screening in the MSU FHB nursery in The third line we have focused on MTHW9420 is a hard white spring wheat with high yield potential under irrigation. MTHW9420 has relatively low protein, thus we truncated the backcrossing program at BC 1 and will follow a single seed descent program to develop materials with white seeds, higher protein, and scab resistance. This project will have inbred materials in the field in We have made additional crosses with numerous FHB-resistant parents, and these materials are included in the standard spring wheat breeding cycle. We have concentrated on the Sumai-3 resistance source, although we also have materials in the program that derive from the dicoccoides resistance found in Howard. Our long-term plan with these crosses is to select for agronomic and quality characteristics first, and then to screen elite lines for resistance. The goal is to be sure that the breeding program has germplasm at all stages of development with FHB resistance. We will have a more extensive set of (Form FPR07) 2

111 PI: Dyer, Alan USDA-ARS Agreement #: materials for testing in These will include superior lines selected from the Choteau and MT0249 backcrosses, along with inbred materials from additional forward crossing projects. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have documented the impact of Scab disease on spring wheat under irrigated conditions in Montana. Growers were also not aware of the high levels of grain DON as documented by the tests conducted from NDSU laboratory. These field trials have convinced growers and seed dealers that FHB resistance was an effective tool in management of scab disease. We have utilized extension networks such as AgAlerts, a MontGuide, and MAES Research & Extension Center Tours to educate the importance of scab disease and the available management tools. Impact: Several varieties with FHB resistance from WestBred, LLC and AgriPro-Coker, Inc. have been widely distributed through their dealer associates. MSU spring wheat program is making rapid progress on FHB resistance with adapted varieties for Montana. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Fusarium head blight in Montana does originate from infested residue and volunteer wheat following grain harvest. The primary inoculum for infection is macroconidia produced from the infested residue and it is not thought that ascospore infection from corn or wheat residue plays a major role. The effective resistance from the Sumai3 QTL under disease conditions and molecular markers have allowed for the backcross of FHB resistance into adapted spring wheat varieties for Montana. Screening the breeding lines and varieties under natural field conditions has been successful. Our goal is the development of scab resistant varieties adapted to sawfly areas and irrigated production. (Form FPR07) 3

112 PI: Dyer, Alan USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. William Grey, Alan Dyer and Luther Talbert Responding to Fusarium head blight for the northern Rocky Mountains and western Great Plains. pg Proceedings of the 2007 National Fusarium Head Blight Forum. The Westin Crown Center, Kansas City, MO, 2-4 December William Grey, Alan Dyer, and Luther Talbert Responding to Fusarium Head Blight and Grain Deoxynivalenol in Montana. Midwest Association of Analytical Communities, International. Midwest Sectional Meeting, Bozeman June 8-10, Mary Burrows, William Grey, and Alan Dyer Fusarium head blight (scab) of wheat and barley. MontGuide, MSU Extension, MT200806AG New 6/08. MSU Press Release Growers gain approval for fungicide to fight wheat and barley scab, 23 May 2008 Radio News Alerts: July 8, 2007: Northern Ag Network, Fusarium head scab risk. April 29, 2008: Northern Ag Network, Current condition of the wheat crop and diseases to be aware of. June 16, 2008: Northern Ag Network, Wheat viruses and chemicals available for head scab control. AgAlerts released and number of hits as of 25 June, 2008 (359 total subscribers): April. Proline registered for use, 117 hits 19 July. Spray to reduce scab risk, 139 hits 23 July. Information available online about Fusarium head scab, 207 hits January. Assess your risk for scab when choosing a spring wheat variety to be planted under irrigated conditions, 151 hits 17 January. Pesticide issues to be aware of in 2008, 187 hits 30 April. Possible fungicide shortages in the United States, 318 hits 22 May. Tebuconazole registered for control of Fusarium head blight (scab) of wheat and barley, 200 hits 16 June. Prosaro labeled for scab control in wheat and barley, 167 hits 16 June. Scab risk in 2008, 152 hits (Form FPR07) 4

113 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Elias Elias Institution: North Dakota State University Address: Department of Plant Sciences 166 Loftsgrad Hall Box 5051 Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 130,732 Identify and Develop Durum Wheat Resistant to Fusarium Head blight. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGR Identify Sources of Resistance to Fusarium Head Blight in Durum Wheat. $38,049 VDUN Development of Durum Wheat Reistant to Fusarium Head Blight. $ 92,683 Total Award Amount $ 130,732 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

114 PI: Elias, Elias USDA-ARS Agreement #: Project 1: Identify Sources of Resistance to Fusarium Head Blight in Durum Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Durum Wheat is very susceptible to Fusarium head blight (FHB) caused by the fungus Fusarium graminearum Schwabe (teleomorph Gibberella Zeae (Schw.) Petch. Sources of resistance to FHB in durum wheat that are equivalent to the Chinese spring wheat Sumai 3 are not available yet. Our objective is to identify sources of resistance that can be utilized by durum plant breeders to develop FHB resistant cultivars. There are 7,000 durum wheat accessions at the National small grain Collection, Aberdeen, ID that are available for evaluating for FHB resistance. We are in the process of evaluating these accessions in field nurseries in China and greenhouses in North Dakota. In addition to these we are evaluating germplasm from the International Center of Agricultural Research in the Dry Areas (ICARDA) and International Maize and Wheat Improvement Center (CIMMYT). 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: To date we have evaluated 7,000 accessions from the world collection. After several evaluations in the field and greenhouses seven accessions maintained disease severity less than 30%. These accessions will be used as parents in crosses. Nine hundred seventy-nine new accessions were evaluated in China. Of these 109 accessions were selected and evaluated for the second time in the greenhouse. Of the 109 accessions 25 were selected for further FHB evaluations in the field and the greenhouse. Twenty-seven lines from crosses with Tunisian lines were selected from Preliminary Yield Trials to be evaluated in Advanced Yield Trials. The diversity study on the Tunisian lines was completed. Tunisian 7 appears to be significantly different than Tunisian 18, 34, 36, and 108. A study to haplotype these sources of resistance has been initiated. Impact: Any resistant germplasm that is identified above could potentially lead into the development of FHB resistant durum cultivars. Resistant durum cultivars will generate million of dollars to the farm economy in the Midwest and will insure the stability of the durum industry in the United States. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The five Tunisian lines and the durum accessions that we have identified to have resistance to FHB are being shared with breeders working on developing FHB resistant durum cultivars. (Form FPR07) 2

115 PI: Elias, Elias USDA-ARS Agreement #: Project 2: Development of Durum Wheat Resistant to Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) caused by the fungus Fusarium graminearum Schwabe (telomorph Gibberella zea (Schwein.) Petch. has been seriously attacking durum wheat. Since 1993, it is estimated that FHB has cost over $3 billion in direct and indirect losses in North Dakota. Although fungicides may reduce FHB, using genetic resistance is the most environmentally safe and economical way to control the disease. The objective of this project is to incorporate identified sources of resistance into the currently susceptible durum wheat germplasm in order to develop resistant cultivars. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The cultivar Divide that have some level of resistance to FHB is gaining acreages in North Dakota. Sumai 3 and Wangshuibai sources of resistance: 6 lines were evaluated in the Uniform Regional Nursery 40 lines were evaluated in the Elite Advanced Yield Trials 56 lines were evaluated in the Advanced Yield Trials 810 lines were evaluated in the Preliminary Yield Trials 58 populations were screened in the field and greenhouses Impact: The above developed material is the only known improved durum germplasm with Fusarium head blight resistance. This germplasm is vital for the survival of the Midwest durum producers. Since the Midwest produces over 75% of the US durum, this germplasm has a major impact on the pasta industry and the US economy. Divide, based on its FHB resistance and yield advantage it could replace up to 30% of the acreage of the current grown cultivars in the Midwest which would generate millions of dollars into the economy. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The producers of North Dakota have a durum cultivar (Divide) that has a moderate level of resistance to FHB. (Form FPR07) 3

116 PI: Elias, Elias USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None (Form FPR07) 4

117 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: James English Institution: University of Missouri Address: 108 Waters Hall Columbia, MO Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 39,024 Selection of Defense Peptides to Protect Wheat from Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * PGG ARS Adjusted Award Project Title Amount Selection of Defense Peptides to Protect Wheat from Fusarium Head $39,024 Blight. Total Award Amount $ 39,024 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

118 PI: English, James USDA-ARS Agreement #: Project 1: Selection of Defense Peptides to Protect Wheat from Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? There is a continuing need for enhanced wheat resistance to scab. In this regard, recently developed biotechnology tools have promise for designing unique new forms of resistance that can complement current wheat breeding efforts. Scab is caused by several Fusarium species, including in particular, F. graminearum. Wheat is susceptible to infection by Fusarium from the time of initial flowering through the soft dough stage of kernel development. Numerous studies have shown that initial infection is caused by germinating ascospores and macroconidia (germlings) of the pathogen. Beyond initial infection, pathogen hyphal growth occurs beneath the cuticle layer of floral parts, and eventually hyphal penetration of cell walls of parenchyma cells leads to cell degradation and development of scab symptoms. This project addresses the need to specifically protect flowers and kernels from Fusarium infection and colonization. We are developing defense peptides that can be deployed in wheat plants for protection of flowers during these early stages of pathogenesis. Peptides are selected from combinatorial phage-display peptide libraries that allow the identification of peptides that bind to multiple cell-surface molecules of the pathogen. Based on experience with several other plant pathogens, some of these binding peptides will disrupt pathogen development and limit disease. This platform peptide technology will complement ongoing resistance breeding that is directed against scab. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The first step in developing defense peptides for scab protection is selection of candidate peptides that bind to Fusarium germlings and disrupt growth and development. We have made initial selections by screening a library of random phage-display peptides for affinity binding with germlings derived from macroconidia. The original library included about one billion peptide variants. Each peptide variant is 8 amino acids in length. Numerous phage-peptide clones were recovered after three rounds of screening and amplification. Evaluation of recovery rates indicated selectivity of phage-peptide clone recovery. A collection of 35 randomly selected clones were sequenced initially. Of these clones, about 25% displayed identical peptide inserts. Additional motifs with sequence redundancy were also identified along with several unique sequences. This pattern of recovery is indicative of appropriate sampling of the initial peptide pool for affinity binding. (Form FPR07) 2

119 PI: English, James USDA-ARS Agreement #: We have performed initial evaluations of the inhibitory potential of affinity-selected, phage-display peptides. In these evaluations, we have found phage-peptide clones that significantly slow germling development in relation to water and non-selected phage library controls. We are also testing selected peptides for inhibitory effects against germlings derived from ascospores. We have begun to incorporate candidate defense peptides into plant-derived protein carrier molecules. These fused peptide-protein scaffolds will be tested for germling inhibition by both in vitro and in planta assays. In addition to developing and testing scaffold-displayed peptides, we have made additional selections of germling-binding peptides from a different library that contains peptides 12-amino acids in length. Recovered peptides are also being assessed for germling inhibition. We are a Impact: We have developed the first set of peptides that have the potential to directly disrupt germling penetration of wheat flowers and kernals, and thus slow or inhibit scab development. Completion of proof of concept for the use of defense peptides will open the door for development of unique wheat germplasm with defensive traits that can enhance existing resistance established by traditional breeding methods. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Genomic and proteomic studies are elucidating what proteins and other factors produced by Fusarium are critical to scab development. These molecules are targets for disruption by application of defense peptides. The platform technologies developed in this project will be made available to the scab research community and enable researchers to develop defense peptides against targets of interest. (Form FPR07) 3

120 PI: English, James USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Poster Presentations Inhibition of Fusarium graminearum germling development caused by combinatorially selected defense peptides. N. W. GROSS (1), Z. D. Fang (1), C. J. Murphy (1), B. Cooper (2), J. T. English (1) (1) Division of Plant Sciences, University of Missouri, Columbia, MO, USA; (2) Soybean Genomics and Improvement Laboratory, USDA- ARS, Beltsville, MD American Phytopathological Society Annual Meeting. San Diego, CA, July Inhibition of Fusarium graminearum germling development caused by combinatorially selected defense peptides. N. W. Gross 1, Z. D. Fang 1, B. Cooper 2, and J. T. English 1. 1 Division of Plant Sciences, University of Missouri, Columbia, MO and 2 Soybean genomics and Improvement Laboratory, USDA-ARS, Beltsville, MD National Fusarium Head Blight Forum. Kansas City, MO, December (Form FPR07) 4

121 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Justin Faris Institution: USDA-ARS Address: NCSL P.O. Box 5677 State Univ. Station Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 18,000 Pyramiding Three Triticum dicoccoides Derived FHB Resistance QTL in Durum Wheat. USWBSI Individual Project(s) USWBSI Research Area * HGR ARS Adjusted Award Project Title Amount Pyramiding Three Triticum dicoccoides Derived FHB Resistance $18,000 QTL in Durum Wheat. Total Award Amount $ 18,000 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

122 PI: Faris, Justin USDA-ARS Agreement #: NA Project 1: Pyramiding Three Triticum dicoccoides Derived FHB Resistance QTL in Durum Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) is one of the most devastating diseases of wheat. Resistant sources of hexaploid bread wheat have been identified and are currently being employed in breeding programs, but development of resistant tetraploid durum wheat has met with less success. Moderate levels of resistance have been identified in Triticum dicoccoides, a wild tetraploid relative, which readily hybridizes with durum wheat. In the T. dicoccoides accession Israel A, QTLs and associated molecular markers for FHB resistance have been identified on chromosomes 3A and 6B. A different accession of T. dicoccoides (PI478742) with moderate levels of FHB resistance was shown to harbor a QTL on chromosome 7A for FHB resistance. Molecular markers linked to these QTL should expedite the pyramiding of T. dicoccoides-derived resistance genes into elite durum lines using marker-assisted selection. Combining the FHB resistance genes from T. dicoccoides chromosomes 3A, 6B, and 7A along with genes identified by others from different sources should lead to a highly resistant durum cultivar. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Initial mapping of the 7A FHB resistance defined the QTL to a 20 cm segment, which is rather large for efficient marker-assisted selection. In this work, we targeted the 7A QTL region with additional PCR-based markers and identified four new markers within the region. The QTL is now delineated to an 8 cm interval by SSR markers, which allow for much more efficient selection of the QTL. We identified two FHB resistant F 3 plants, designated and 56-78, that were homozygous for the 3A and 6B QTLs in the Langdon background. These plants were crossed to LD7A-28, which is a recombinant line homozygous for the 7A QTL. The resulting F 1 plants, which were heterozygous for all three QTL, were then crossed to the durum variety Divide. A total of about 500 plants from these crosses were genotyped with molecular markers for the 3A, 6B, and 7A QTLs and four progeny heterozygous for all three QTL were identified. These plants will be crossed again to Divide to recover more of the Divide background, and the progeny will again be genotyped to retain plants with all three FHB resistance QTL. Impact: Incorporation of the 7A QTL identified in this research together with the 3A and 6B QTLs will lead to higher levels of FHB resistance in tetraploid wheat. The additional markers associated with the 7A QTL provide valuable tools for breeders and geneticists to use in marker-assisted selection schemes for the development of FHB resistant germplasm and varieties. The pyramiding of the 3A, 6B, and 7A T. dicoccoides derived QTLs into the (Form FPR07) 2

123 PI: Faris, Justin USDA-ARS Agreement #: NA Divide background provides the first step toward the development of an FHB resistant durum variety. The molecular markers associated with each QTL make the pyramiding of the QTLs and recovery of the Divide background extremely efficient compared to having to conduct phenotypic evaluations. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The new markers identified for the 7A FHB resistance QTL are valuable for breeders and geneticists who wish to introgress the 7A QTL into adapted material. These markers are currently being used by the USDA-ARS genotyping lab in Fargo, ND and by the North Dakota durum breeder. The community now has tetraploid germplasm containing the 3A, 6B, and 7A FHB resistance QTLs in a single background. Only further recovery of the recurrent parent background is needed before an adapted FHB resistant durum germplasm is available. (Form FPR07) 3

124 PI: Faris, Justin USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None. (Form FPR07) 4

125 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Liane Gale Institution: University of Minnesota Address: Department of Plant Pathology 200 Oak Street S.E. Minneapolis, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 36,219 Heterogeneity & Toxigenic Potential of U.S. Fusarium graminearum. USWBSI Individual Project(s) USWBSI Research Area * PGG ARS Adjusted Award Project Title Amount Heterogeneity and Toxigenic Potential of U.S. Fusarium $36,219 graminearum Population. Total Award Amount $ 36,219 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

126 PI: Gale, Liane USDA-ARS Agreement #: Project 1: Heterogeneity and Toxigenic Potential of U.S. Fusarium graminearum Population. 1. What major problem or issue is being resolved and how are you resolving it? Our long term objectives are to accurately determine the composition of genetically coherent populations of Fusarium Head Blight (FHB) pathogens in economically important wheat producing areas of the world with special focus on the U.S., to determine their genetic structure, to evaluate their potential to change in composition and genetic structure and to determine the effect of such change on deployed host genotypes and/or other agricultural practices. Our USWBSI-funded research established that U.S. isolates of Fusarium graminearum, the main causal agent of FHB in small grains, do not belong simply in a single, homogeneous and interbreeding population as has been previously assumed, but that the pathogen population composition in the U.S. is both complex and in flux. In addition to the widespread F. graminearum population (Midwestern 15ADON population) we have identified, molecularly and phenotypically characterized, and geographically and temporally mapped F. graminearum populations that are genetically distinct from the MW 15ADON population. Genetically distinct populations have been identified in the Upper Midwestern U.S. (MN, ND, and more recently in SD) and in Louisiana. We also learned that differences between populations are not only present at a molecular level, but also at a phenotypic level, affecting traits that are agriculturally and economically important. Briefly, representatives of emerging populations of F. graminearum that have been identified in the Upper Midwestern U.S. have a higher toxigenic potential than the MW 15ADON population as they produce substantially more deoxynivalenol (DON) on wheat in greenhouse experiments than members of the MW15ADON population. Members of F. graminearum populations in Louisiana were found to be predominantly nivalenol producers. Nivalenol is believed to be several times more toxic than DON. Prior to our efforts, it was assumed that nivalenol producers of F. graminearum in the U.S. are rare or non-existent. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment #1: We demonstrated that the emergent populations of F. graminearum that have a higher toxigenic potential than the common MW15DON population and that were first identified from samples in ND and MN, are moving further south. Population genetic analysis of 1,132 isolates from a 2006 collection found members of the emergent populations in areas were these populations were previously rare or absent. Impact: For the first time, isolates of F. graminearum with a 3ADON trichothecene type were identified in South Dakota. The isolates belong to the highly toxigenic Upper Midwestern (UMW) 3ADON population and were detected in 4 out of 10 counties sampled in South Dakota. Accurate knowledge of the population composition of F. graminearum is important for breeders, field pathologists and farmers alike as pathogen variability may affect the reaction of host genotypes that are either deployed or in development and/or the efficacy of other agricultural practices (e.g. chemical control by fungicides). As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: An update on the population composition of F. graminearum that is both complex and in flux in sampled regions of North Dakota, South Dakota and Minnesota. Evidence that highly toxigenic emergent populations are moving further south. Accomplishment #2: Previously, we demonstrated that F. graminearum isolates from Louisiana are predominantly nivalenol producers. As most isolates from initial collections were sampled (Form FPR07) 2

127 PI: Gale, Liane USDA-ARS Agreement #: from nurseries, it was not clear whether commercial fields would harbor nivalenol producers as well. We found that isolates of F. graminearum from commercial fields in Louisiana are also predominantly (~80%) nivalenol producers. Nivalenol producers of F. graminearum were also identified from Arkansas, though examined isolates were from older collections (early 1990s). Development of PCR-RFLP markers and subsequent population genetic analyses further indicated that isolates from Louisiana largely belong to one of three genetically distinct populations. In contrast, the majority of F. graminearum isolates from Arkansas could not be grouped into specific populations. This was taken as an indication that Arkansas may constitute a hybrid zone between the Midwestern and southern populations of F. graminearum. Impact: For the first time, significant numbers of F. graminearum isolates from commercial fields in Louisiana were examined for their trichothecene type. As previous work predominantly had examined isolates from nurseries, we established a more complete picture of population composition in Louisiana. Some of our characterized isolates were given to collaborator Dr. Gene Milus for further use in greenhouse experiments. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Information that nivalenol producers of F. graminearum are omnipresent in Southern Louisiana and that nivalenol producers also occur in Arkansas. Accomplishment #3: We determined in greenhouse experiments that members of the emergent populations have a higher toxigenic potential on popular spring wheat cultivars. For example, members of UMW 3ADON population produced on average 87% (range 40%-128%) more DON on tested cultivars (Alsen, Knudson, Briggs, Oklee, Granite, Freyr) than members of the MW 15ADON population. Impact: For the first time it was established that the increased toxigenic potential of the emergent populations is also observed for commercially important cultivars. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: A working hypothesis that recent changes in population composition in the Upper Midwest may have resulted to an increase of DON levels in small grains even though the level of disease may not have increased. (Form FPR07) 3

128 PI: Gale, Liane USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-reviewed publications: Suga, H., Karugia, G. W., Ward, T., Gale, L. R., Tomimura, K., Nakajima, T., Miyasaka, A., Koizumi, S., Kageyama,, K., and Hyakumachi, M. (2008). Molecular characterization of the Fusarium graminearum species complex in Japan. Phytopathology 98: Gale, L. R., Ward,T. J., Balmas,V., and Kistler, H. C. (2007). Population subdivision of Fusarium graminearum sensu stricto in the Upper Midwestern United States. Phytopathology 97: Starkey, D. E., Ward, T. J., Aoki, T., Gale, L. R., Kistler, H. C., Geiser, D. M., Suga, H., Toth, B., Varga, J. and O'Donnell, K. (2007). Global molecular surveillance reveals novel Fusarium head blight species and trichothecene toxin diversity. Fungal Genetics and Biology 44: Presentations: Phenotypic and Molecular Diversity of Fusarium graminearum sensu stricto from the U.S. National Fusarium Head Blight Forum, Kansas City, MO, Non-peer reviewed articles: Gale, L. R., and Kistler, H. C. (2007). Phenotypic and molecular diversity of Fusarium graminearum sensu stricto from the U.S. Page 27 in 2006 National Fusarium Head Blight Forum Proceedings. Gale, L. R., Harrison, S. A., Milus, E. A., Ochocki, J. E., O Donnell, K. O., Ward, T. J., and Kistler, H. C. (2007). Diversity in Fusarium graminearum sensu stricto from the U.S.: an update. Page 26 in 2006 National Fusarium Head Blight Forum Proceedings. (Form FPR07) 4

129 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: David Garvin Institution: USDA-ARS Address: Room 411 Borlaug Hall 1991 Upper Buford Circle St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 6,679 Coordination of a Uniform Regional Scab Nursery for Spring Wheat Parents. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Coordination of a Uniform Regional Scab Nursery for Spring Wheat $6679 Parents. Total Award Amount $ 6,679 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

130 PI: Garvin, David USDA-ARS Agreement #: NA Project 1: Coordination of a Uniform Regional Scab Nursery for Spring Wheat Parents. 1. What major problem or issue is being resolved and how are you resolving it? The significant environmental effect associated with the wheat disease scab (Fusarium head blight) poses an obstacle for breeding programs seeking to develop more scab-resistant germplasm. Undertaking multisite evaluations across a range of environments is the only way to adequately assess scab resistance in germplasm. The Uniform Regional Scab Nursery for Spring Wheat Parents (URSN) seeks to address this need. Interested parties from academia and private companies nominate germplasm for scab resistance evaluation at field locations in Minnesota, North Dakota, South Dakota, and Canada that provide conditions to enhance scab development on wheat. Both hard red spring wheat and durum wheat are included in the URSN. These data are supplied to the coordinator, who oversees its collation and statistical analysis, and produces an annual report for the nursery program. An additional aspect of the URSN is to encourage open and free germplasm exchange, in order to foster cooperation among breeders in efforts to develop scab resistant germplasm. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The URSN was grown for the 13th year in 2007, at locations in the U.S. and Canada. Entries were contributed by university, industry, and national wheat breeding programs. Scab resistance-related trait data from the locations were compiled and analyzed, and the annual report that was produced provides individual location data summaries, and data summariesand rankings across locations. Impact: This nursery continues to function as an excellent source of data on wheat scab resistance from the field and as a vehicle for exchange of germplasm among spring wheat breeders in the Upper Midwest. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The 2007 annual report provides readily accessible information on scab resistance in germplasm emerging from a number of wheat breeding programs. This in turn provides guidance to wheat breeders for future parental selections in their crossing programs. No other comparable vehicle for multisite scab evaluations and subsequent germplam exchange exists for the spring wheat region. (Form FPR07) 2

131 PI: Garvin, David USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. This project is designed specifically to produce a non-peer reviewed annual report for distribution to Nursery cooperators and other interested parties. GARVIN, D.F, AND BLANKENHEIM, Z Report of the 2007 Uniform Regional Scab Nursery for Spring Wheat Parents. The report is made available at the USWBSI web site ( and through the Graingenes web site ( (Form FPR07) 3

132 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Bikram Gill Institution: Kansas State University Address: Department of Plant Pathology 4024 Throckmorton Manhattan, KS Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 29,268 Alien Chromosome Engineering and the Deployment of a Novel Source of Fusarium Head Blight Resistance in Wheat. USWBSI Individual Project(s) USWBSI Research Area * HGR ARS Adjusted Award Project Title Amount Alien Chromosome Engineering and the Deployment of a Novel $29,268 Source of Fusarium Head Blight Resistance in Wheat. Total Award Amount $ 29,268 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

133 PI: Gill, Bikram USDA-ARS Agreement #: Project 1: Alien Chromosome Engineering and the Deployment of a Novel Source of Fusarium Head Blight Resistance in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? A potential hazard to FHB resistance breeding is that only a few sources, mainly Sumai 3 and its derivatives, are now widely used around the globe. Thus, additional sources of resistance are needed to broaden the genetic basis of FHB resistance. FHB resistance was identified in the alien species Leymus racemosus (syn. Elymus giganteus). L. racemosus (Lr) chromosomes were isolated as single chromosome additions to bread wheat. Chromosome engineering of these lines was attempted and nine wheat-leymus translocation lines were produced, five using irradiation treatment, and four using Gc gene action (Chen et al. 2005). Three lines were reported to have resistance to FHB. However, all translocations were of non-compensating type, leading to duplications/deficiencies and, hence, may not be deployed in agriculture. We further analyzed a total 58 wheat-leymus introgression lines for their resistance to FHB disease. We discovered a genetically compensating translocation involving the long arm of wheat chromosome 7A and the short arm of Leymus chromosome 7Lr#1 (T7AL 7Lr#1S) conferring resistance to FHB. This translocation may be directly used in breeding programs. Our studies also revealed that three previously reported wheat-leymus non-compensating translocations involved the same Leymus chromosome arm-7lr#1s. A new FHB resistance gene located in the short arm of chromosome 7Lr#1 was named as Fhb3. Currently, we are using chromosome engineering to reduce linkage drag of the Leymus 7Lr#1S segment in wheat background. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A wheat-leymus translocation line (T09) conferring FHB resistance was identified as a genetically compensating translocation involving the long arm of wheat chromosome 7A and the short arm of Leymus chromosome 7Lr#1 (T7AL 7Lr#1S) by C- banding, genomic in situ hybridization, and RFLP analyses. Another two resistant wheat- Leymus introgression lines (T01 and T14) were identified as double translocations. Lines T01 (2n = 44) and T14 (2n = 44) each had two pairs of independent translocation chromosomes. T01 had T4BS 4BL-7Lr#1S + T4BL-7Lr#1S 5Lr#1S. T14 had T6BS 6BL- 7Lr#1S + T6BL 5Lr#1S. The three translocation lines, T01, T09, and T14, and the disomic addition 7Lr#1 were consistently resistant to FHB in greenhouse point-inoculation experiments, whereas the disomic addition 5Lr#1 was susceptible. The data indicated that a novel FHB resistance gene from Leymus, designated Fhb3, resides in the distal region of the short arm of chromosome 7Lr#1, because the resistant translocation lines share a common distal segment of 7Lr#1S. Three PCR-based markers, BE STS, BE STS, and BE STS, specific for 7Lr#1S were developed to expedite marker-assisted selection in breeding programs. The compensating translocation T7AL 7Lr#1S was backcrossed twice to adapted wheat cultivars Overley and Jagger. A total of 192 progeny homozygous for translocation T7AL 7Lr#1S were selected by molecular markers from 436 BC 1 F 2 plants and they will be evaluated for FHB resistance and its effect on DON accumulation in a field nursery in Fall (Form FPR07) 2

134 PI: Gill, Bikram USDA-ARS Agreement #: Chromosome engineering was initiated to reduce genetic linkage drag associated with translocation T7AL 7Lr#1S. This line was crossed twice with the ph1b mutant stock. 154 BC 1 plants were screened using molecular markers to assay for ph1b and T7AL 7Lr#1S. Sixty-one plants were found to be homozygous ph1b/ph1b and heterozygous for the translocation chromosome T7AL 7Lr#1S/7A. These plants were either backcrossed with Overley and Danby or selfed. Large recombinant populations of 1,400 BC 2 seeds and more than 8,000 BC 1 F 2 seeds were developed. In homozygous ph1b genotypes, the alien 7Lr#1S arm with Fhb3 is expected to recombine with homoeologous wheat arm 7AS. Work is ongoing to identify wheat-leymus recombinants with smaller alien segments that still retain the FHB resistance gene Fhb3. Impact: Fhb3 is the first FHB resistance gene identified from an alien species, Leymus racemosus. This is a novel gene different from previously reported FHB resistance genes in Triticum aestivum, providing a new source for gene pyramiding in breeding programs. The advantage of the translocation T7AL.7Lr#1S is its large phenotypic effect and simple inheritance thereby facilitating its rapid deployment in wheat breeding programs. Three PCR-based markers developed will facilitate marker-assisted selection. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: 1. Breeders have an additional source of FHB resistance carrying Fhb3 that was not previously known or well-characterized. 2. Molecular markers to assay Fhb3 gene in breeding programs. 3. Prebreeding of Fhb3 with elite wheats. 4. Publication of a refereed paper describing the methodology of alien introgression. 5. The advanced germplasm will be released to plant breeders. The published data represent an important technology transfer because wheat geneticists and breeders can use the germplasm and associated markers in breeding programs. (Form FPR07) 3

135 PI: Gill, Bikram USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. L.L. Qi, M.O. Pumphrey, B. Friebe, P.D. Chen and B.S. Gill, 2008 Molecular cytogenetic characterization of alien introgressions with gene Fhb3 for resistance to Fusarium head blight disease of wheat. Theor Appl Genet (in press) L.L. Qi, M.O. Pumphrey, B. Friebe, P.D. Chen and B.S. Gill, 2007 Molecular characterization of a wheat-leymus compensating translocation line conferring resistance to Fusarium head blight. Proceeding of 2007 National Fusarium Head Blight Forum p: 225 (Form FPR07) 4

136 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Karl Glover Institution: South Dakota State University Address: Plant Science Department Box 2207A NPB 247 Brookings, SD Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Spring Wheat Breeding for Scab Resistance in South Dakota. Title: FY07 ARS Award Amount: $ 69,681 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount VDUN Spring Wheat Breeding for Scab Resistance in South Dakota. $69,681 Total Award Amount $ 69,681 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

137 PI: Glover, Karl USDA-ARS Agreement #: Project 1: Spring Wheat Breeding for Scab Resistance in South Dakota. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) is a serious wheat disease that continues to pose as a production threat within South Dakota as well as the North Central region of the USA. In an attempt to alleviate this threat to wheat production, development of resistant varieties has became a high priority within the spring wheat breeding program at South Dakota State University. An aggressive program was initiated to accelerate the development of spring wheat varieties with improved FHB resistance and desirable agronomic traits. Established off-season nurseries and mist-irrigated greenhouse and field screening nurseries are utilized to accelerate breeding efforts in improving resistance along with desirable agronomic characteristics. Three early generations of breeding materials are evaluated for scab resistance each year: two generations in the greenhouse and one in the field. Approximately 8,000 individual hills are evaluated in the greenhouse nurseries and 3,000 rows are screened in the field nurseries. Both the field and greenhouse nurseries are inoculated with infested corn and conidial suspensions. A mistirrigation system is used to provide a favorable environment for infection and disease development. Each year we make a large number of crosses to introduce new resistance genes and create new resistance gene combinations. Sources of resistance used in the crosses include materials from the Uniform Regional Scab Nursery (URSN) for spring wheat parents, (a cooperative regional effort to identify and utilize sources of scab resistance) newly identified germplasm provided through introduction and evaluation efforts, other introduced sources, as well as both varieties and advanced breeding lines with various levels of resistance. The offseason nursery aids in the simultaneous selection for resistance and desirable agronomic characteristics. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Our program has provided elevated levels of resistance to FHB in the form of Hard Red Spring Wheat (HRSW) varieties and germplasm made available to regional growers and other breeders that wish to utilize the germplasm Within the past several years, three varieties have been released to growers by our program. We anticipate the release of a fourth prior to the 2009 growing season. Over three years of simultaneous testing, the FHB disease index ratings recorded for Briggs, Granger, Traverse, and SD3851 were 39.4, 34.7, 31.6, and 20.8, respectively, compared to Sumai 3, (15.4) the resistant check. Impact: Elevated resistance levels in released varieties are immediately utilized by the most apparent benefactors of our work; HRSW producers. Through utilizing the elevated resistance levels, growers are more able to protect themselves from suffering complete devastation of fields in the presence of a severe FHB epidemic. Elevated resistance levels in germplasm is also quite often utilized by a less immediately apparent benefactor group; HRSW breeders. Through utilizing both germplasm and released varieties, other breeding programs strive to further (Form FPR07) 2

138 PI: Glover, Karl USDA-ARS Agreement #: increase FHB resistance among germplasm pools that will eventually result in the release of continually improved varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Our efforts have led to the release of three varieties since 2002 that possess some FHB resistance. Prior to the 2009 growing season, a fourth release is planned. This experimental line, (SD3851) has shown itself to be nearly as resistant as Sumia 3 in our replicated tests and is agronomically far superior. These accomplishments have led to the release of several HRSW varieties that possess some resistance to FHB and are specifically adapted to South Dakota and the northern Great Plains region. (Form FPR07) 3

139 PI: Glover, Karl USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-reviewed articles M. Kadariya, K. D. Glover, M. Mergoum, and L. E. Osborne. 200X. Biplot Analysis of Agronomic and Fusarium Head Blight Resistance Traits in Spring Wheat. J. of Crop Improvement. (Accepted). Poster/abstract presentations Lawrence E. Osborne, Jeffery M. Stein, Karl D. Glover, and Christopher A. Nelson Spore Load, Disease, and DON: An Inoculum Gradient Study using Sister Wheat Lines. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the 2007 National Fusarium Head Blight Furum; 2007, 2-4 December; Kansas City, MO, USA. Michigan State University, East Lansing, MI. p K.R. Ruden, B.E. Ruden, K.D. Glover, and J.L. Kleinjan Uniform Fungicide Performance Trials for the Suppression of Fusarium Head Blight in South Dakota. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the 2007 National Fusarium Head Blight Furum; 2007, 2-4 December; Kansas City, MO, USA. Michigan State University, East Lansing, MI. p Bhoja Raj Basnet, Yang Yen, Shiaoman Chao,.and Karl D. Glover Genetic Linkage Mapping with DArT Markers to Detect Scab Resistance QTLs in a Sumai-3 Derived Wheat Population. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the 2007 National Fusarium Head Blight Furum; 2007, 2-4 December; Kansas City, MO, USA. Michigan State University, East Lansing, MI. p S. Malla, A.M.H. Ibrahim,.and K. Glover Winter and Spring Wheat Parental Diallel Anaysis for Scab Resistance. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the 2007 National Fusarium Head Blight Furum; 2007, 2-4 December; Kansas City, MO, USA. Michigan State University, East Lansing, MI. p Rosarya, R.U., Maxson-Stein K.L., Glover, K.D., Stein, J.M., and Gonzalez-Hernandez, J.L Family Based Mapping of FHB Resistance QTLs in Hexaploid Wheat. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), Proceedings of the 2007 National Fusarium Head Blight Furum; 2007, 2-4 December; Kansas City, MO, USA. Michigan State University, East Lansing, MI. p (Form FPR07) 4

140 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Arvydas Grybauskas Institution: University of Maryland Address: Dept. of Plant Science and Landscape Architecture 2102 Plant Sci. Bldg. 036 College Park, MD Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Integrated Management of Fusarium Head Blight in Maryland. Title: FY07 ARS Award Amount: $ 15,366 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Integrated Management of Fusarium Head Blight in Maryland. $15,366 Total Award Amount $ 15,366 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

141 PI: Grybauskas, Arvydas USDA-ARS Agreement #: Project 1: Integrated Management of Fusarium Head Blight in Maryland. 1. What major problem or issue is being resolved and how are you resolving it? The search for management tactics that can protect producers from the losses in yield and mycotoxin contamination associated with Fusarium head blight has taken great strides forward with the development of cultivars with some resistance and new triazole fungicides. However, neither tactic alone has proved to be adequate in seasons highly favorable for disease development. Furthermore, some of the more highly resistant cultivars have had lower yield potentials than highly adapted but susceptible cultivars and thus lose favor after a season with low disease development. The effectiveness of fungicides has also been inadequate under certain conditions especially if relied upon as a sole management tactic. An integrated approach primarily testing the combination of cultivars that have a modicum of resistance with the best fungicide under various debris management / rotation schemes is being tested and demonstrated in this project. We are also using the web-based forecasting system to help producers determine the real-time risk of disease development to help determine if the fungicide application is warranted that season. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The environmental conditions were not favorable for a serious epidemic of Fusarium head blight in Maryland in Our program included in the project a trial that was inoculated and mist-irrigated that produced disease conditions and provided an excellent proof of concept and demonstration of the effectiveness of a combination of moderate resistance with a moderately effective fungicide. In particular, under moderate disease conditions, resistance and fungicides reduced deoxynivalenol in an additive manner. The results from uninoculated trials provided data to the forecasting project. Impact: These results establish that the combination of tactics can reduce mycotoxin accumulation to a greater extent than either tactic used alone. It also helps to establish a strategy that a high yield potential cultivar with a only a moderate level of resistance may be effectively managed with fungicides in seasons favorable for disease development without the yield drag cost associated with a cultivar having a higher level of resistance. This is a critical step in gaining grower acceptance particularly within the region and encourages the selection of cultivars with relatively minor levels of resistance. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We now have data and regional demonstration of the effectiveness of integration of management practices on DON reduction. (Form FPR07) 2

142 PI: Grybauskas, Arvydas USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Grybauskas, A. P Fusarium head blight of small grains. Southern Association of Seed Certification and Technology. April 12, 2007, Annapolis, MD. Grybauskas, A.P. and E. Reed MD Field Crop Disease Management Research Update. NJ-DelMarVa-Pa Plant Pathologist Assoc., March 3, 2008, Newark, De. Grybauskas, A.P Field crop disease management trials in Maryland. APS Potomac division annual meeting, Extension/Industry session, March 21-23, 2007, Blacksburg, VA. Fungicide issues in field crops in A. Grybauskas. Queen Anne s County Agronomy Day, Centreville MD, March 7, Field crop disease management for A. Grybauskas. Montgomery and Howard County Agronomy Update, Derwood MD, February 27, Have you heard this one: prices are up protect your investments? A. Grybauskas. Maryland Pesticide Conference, Denton MD, February 7, Have you heard this one: prices are up protect your investments? A. Grybauskas. Maryland Pesticide Conference, Frederick MD, February 5, Wheat disease prospects after the drought of 07 & Soybean rust status. A. Grybauskas. Queen Anne County Small Grain Meeting, August 27, 2007, Centreville, MD. Small grain diseases in 2007 and Soybean Rust status. A. Grybauskas. Wye Grains Twilight Tour, May 24, 2007, Queenstown, MD. Field Crop Disease Management Update. A. Grybauskas. Harford County Mid-Winter Meeting, Rocks, MD, February 22, Disease Management update. A. Grybauskas, Montgomery County Winter Agronomy meeting, Derwood, MD, February 21, Disease Management update. A. Grybauskas, Cecil County Winter meeting, Rising Sun, MD, February 20, Wheat Scab and Soybean rust current research and outlook. A. Grybauskas. Maryland Crop Improvement Association 100 th Anniversary meeting, Ruthsburg, MD, February 8, Field Crop Disease Management Update. A. Grybauskas. Dorchester County Winter Program, Hurlock, MD, January 25, (Form FPR07) 3

143 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Scott Halley Institution: North Dakota State University Address: Langdon Research Extension Center th Ave NE Langdon, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 19,512 Aerial and Ground Spray Application Technology for Enhanced Fungicide Efficacy for Control of FHB. USWBSI Individual Project(s) USWBSI Research Area * CBCC ARS Adjusted Award Project Title Amount Spray Application Technology Evaluations for Enhanced Fungicide $19,512 Efficacy. Total Award Amount $ 19,512 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

144 PI: Halley, Scott USDA-ARS Agreement #: Project 1: Spray Application Technology Evaluations for Enhanced Fungicide Efficacy. 1. What major problem or issue is being resolved and how are you resolving it? Fungicide is applied to small grains for control of Fusarium head blight (FHB) using ground application equipment that has two distinct delivery methods. One system, a hydraulic delivery system, has shown to be a good spray delivery method which has been studied and reported. The second system is an air-spray system which uses a high velocity air system to carry the spray to the target. The parameters tested include air delivery speed, drop size and orifice delivery angle relative to the orientation of the grain head. This research project is studying the most efficacious delivery parameter combinations with the air delivery system. The studies are designed to test the results with a factorial arrangement and replication so each individual parameter is evaluated with all other parameter options or combinations. This project will determine barley and hard red spring wheat grain head coverage, fungicide efficacy for reducing effects of FHB on head disease and deoxynivalenol accumulation in the grain and yield and other grain quality parameters. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The first year of the studies showed that several delivery parameter options could effectively improve fungicide efficacy but all options were not equal. Two additional years of study on barley and hard red spring wheat are planned. Impact: Small grain producers will improve economic return and the public will have a safer product to consume. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: When the additional two years of study is complete growers who use an air delivery system will be able to maximize effectiveness of fungicide application at minimal cost and the consumer will have a better and safer product. (Form FPR07) 2

145 PI: Halley, Scott USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. A poster was presented at the USWBSI forum in Kansas City, Missouri and at the North Dakota State Barley Show in Osnabrock, North Dakota. A presentation was also given to the public at the North Dakota State Barley Show and at the Lake Region Roundup Devils Lake, North Dakota. Estimated participants at the two forums were Publications: Aerial Application of Fungicide for the Suppression of Fusarium Head Blight in Small Grains. Publication No. AE May Hofman, V., S. Halley, G. Van Ee, C. Hollingsworth, M. McMullen, and B. Ruden. North Dakota State University Extension Service, Fargo, ND Fungicide Deposition Measurement by Spray Volume, Drop Size, and Sprayer System in Cereal Grains. Halley S., G. Van Ee, V. Hofman, S. Panigrahi, H. Gu American Society of Agricultural and Biological Engineers, Applied Engineering in Agriculture Vol. 24(1): (Form FPR07) 3

146 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Stephen Harrison Institution: Louisiana State University Address: Agronomy Department 104 Sturgis Hall Baton Rouge, LA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 32,213 Development of FHB Resistant Wheat Genotypes Adapted to the Gulf Coast. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Development of FHB Resistant Wheat Genotypes Adapted to the $32,213 Gulf Coast. Total Award Amount $ 32,213 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

147 PI: Harrison, Stephen USDA-ARS Agreement #: Project 1: Development of FHB Resistant Wheat Genotypes Adapted to the Gulf Coast. 1. What major problem or issue is being resolved and how are you resolving it? This project is developing wheat varieties adapted to the Gulf Coast that are resistant to Fusarium Head Blight (FHB), and evaluating FHB reaction of available varieties. The climate and disease spectrum of Louisiana are unique and most varieties developed outside of the region perform poorly. The LSU AgCenter wheat breeding program and its Sungrains partners (Universities of Florida, Georgia, Clemson, and NC State) release high-yielding disease-resistant varieties that account for most of the wheat acreage in the Gulf Coast and Southeastern states. It is important that these programs develop and release highly productive, scab resistant varieties that are embraced and produced by growers. Louisiana FHB populations are unique and 78% were found to be NIV producers. Since NIV is more toxic than DON it is important to develop resistant varieties to prevent movement of NIV contaminated grain into the Mississippi River export elevators. Scab occurs occasionally across Louisiana and frequently in the rice region of southwest Louisiana. Objectives will be accomplished by: (1) participating in regional screening nurseries, (2) evaluating entries in statewide variety trials and uniform nurseries for FHB resistance, and, (3) conducting a breeding program to develop elite varieties with local adaptation and resistance to FHB. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The wheat breeding program continued to make significant progress towards release of locally-adapted varieties with resistance to FHB during Nineteen of 26 breeding lines evaluated in a two-location yield trial, and in three-misted nurseries, were found to contain markers indicating the presence of one or more resistance alleles, including 5AS and 3BS. Data were collected on agronomic traits, FHB resistance, mycotoxin accumulation, and reaction to other diseases. The statewide variety trials should have FHB resistant entries, advanced from these trials, for the first time in LA01164, a topcross derivative from Futai8944, contains 3BS and 5AS QTLs, shows excellent FHB resistance in screening trials, and appears to be well adapted and productive. LA01162 also performed very well and shows good FHB resistance. Sixteen lines were selected for advance, pending complete FHB data, out of a set of about 120 observation plots derived from FHB-targeted crosses. These lines were haplotyped, screened in misted nurseries, and selected for adaptation and general disease resistance. A number of these contain 3BS or 5AS QTLS, while others have resistant parents with less characterized sources in their background. Sixty-four advanced generation headrows were harvest in May, 2008 for advancement to yield plots and misted FHB nurseries. All have known FHB resistant parents in their background. (Form FPR07) 2

148 PI: Harrison, Stephen USDA-ARS Agreement #: Approximately 300 single (widely spaced) plants were harvested from 23 F3 FHB populations and will be also be evaluated in yield trials during Each of these will be genotyped. Plants from two additional populations (from Paul Murphy) that contained sources of FHB resistance from Frontana and 3A-diccocoides were individually selected for plant type and disease reaction. Tissue samples have been sent to the Genotyping Lab and seed of selected pants that contain desired QTL will be shared with other Sungrains breeders this summer. Heads were selected from F2 F4 populations with known FHB resistance and advanced as headrows. Ninety of 280 new crosses made in 2008 contained a known source of FHB resistance and the sources used were diverse and considerably more adapted than those used in initial FHB crosses. In addition to breeding and screening activities, a trial was conducted at two locations to evaluate the interaction between genetic resistance to FHB and chemical control. This replicated, inoculated yield trial included six varieties with different levels of FHB resistance in a factorial arrangement with three levels of fungicide. Data from this and similar trials will allow appropriate site-specific management strategies to be developed for combinations of resistance and disease pressure. Impact: This program is primarily focused on the development of productive varieties with good disease-resistance that are adapted to the Gulf Coast region. The LSUAC wheat breeding program has released four varieties since 2002 and accounted for a large proportion of wheat production in Louisiana during There are no FHB-resistant varieties currently grown in Louisiana. The development of FHB resistant varieties will positively impact wheat production economics and should allow growers in the distressed rice producing region of southwest Louisiana where FHB is a significant problem to expand wheat production. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Louisiana growers will soon have available varieties that are locally adapted and are resistant to Fusarium headlight. There are few cropping options for the rice region and scab-resistant wheat would provide an economically viable option. (Form FPR07) 3

149 PI: Harrison, Stephen USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Gale. Liane R., S.A. Harrison, et al Diversity of Fusarium graminearum sensu strict from the U.S.: An Update. Poster presented at the 2007 FHB meeting, Indianapolis, IN. (Form FPR07) 4

150 USDA-ARS/ U.S. Wheat and Barley Scab Initiative FY07 Preliminary Final Performance Report (approx. May 07 April 08) July 15, 2008 Cover Page PI: Suzanne Hendrich Institution: Iowa State University Address: Food Science and Human Nutrition 220 MacKay Ames, IA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 41,980 Biomarkers of Low Dose Immunotoxicity of Deoxynivalenol in Mice. USWBSI Individual Project(s) USWBSI Research Area * FSTU-R ARS Adjusted Project Title Award Amount Biomarkers of Low Dose Exposure to Deoxynivalenol in Mice. $41,980 Total Award Amount $ 41,980 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form PFPR07)

151 PI: Hendrich, Suzanne USDA-ARS Agreement #: FY07 Preliminary Final Performance Report Project 1: Biomarkers of Low Dose Exposure to Deoxynivalenol in Mice. 1. What major problem or issue is being resolved and how are you resolving it? There is little information available regarding the toxicity of deoxynivalenol (DON), the fungal toxin associated with wheat/barley scab and most commonly found in those grains, especially in doses close to the current voluntary action levels of 0.5 ppm (EU) or 1.0 ppm (US) in finished foods for human intake. Biomarkers for exposure to DON and more evidence regarding NOAELs and LOAELs (no and lowest observed adverse effect levels) for DON is urgently needed. We are addressing this need with mouse model feeding studies within the range of 0-2 ppm DON, observing multiple cell surface markers of leukocytes in peripheral blood in young and old adult mice over 14 and 28 days of DON exposure as a means of assessing toxic effects of DON related to immune function. The rationale for this study design is that this time frame is recognized as a standard for immunotoxicity testing. Additionally, aging is known to suppress some aspects of human immune function, so an immunotoxic fungal toxin such as DON might be likely to cause further suppression, and is an important aspect of investigation to protect vulnerable populations. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: To find biomarkers of the grain fungal toxin, deoxynivalenol (DON), 8 w and 16 mo old male and female BALB/c mice were fed 0, 1 or 2 ppm DON for 14 and 28 d. We hypothesized immunotoxicity in BALB/c mice was greater with aging combined with deoxynivalenol (DON) exposure. By flow cytometry, in young female mice, the percentage of T helper cells in peripheral blood was inhibited at 2.0 ppm after 14 d, not after 28 d; integrin expression (VLA-4 and LFA-1) in neutrophils was inhibited at 2.0 ppm after 28 d; percentage of splenic macrophages were inhibited at 2.0 ppm after 14 d and 28 d. In young male mice, CCR7 expression that directs cells to the lymph nodes was inhibited in blood T helper cells at 2.0 ppm DON after 14 d and 28 d. In old female mice, percentage of CXCR5 + B cells in the blood was inhibited at 1.0 and 2.0 ppm DON after 14 d, but that effect disappeared after 28 d (CXCR5 directs cells to the spleen, the main site of action of mature B cells); the percentage of T cytotoxic cells (CD8+) in blood was inhibited at 2.0 ppm DON after 28 d. Compared with the other three groups, old male mice had the greatest change in leukocyte surface markers: in blood, increased integrins in neutrophils were observed at 1.0 and 2.0 ppm DON after 14 d; and increased CCR9+ T cytotoxic cells targeting small intestine and decreased numbers of T cytotoxic cells were observed at 2.0 ppm DON after 28 d. These results suggest small intestinal inflammatory damage in old male mice exposed to DON. CCR10+ T helper cells and T cytotoxic cells were increased in male mice fed 1.0 ppm DON after 14 d, which suggested low dose of DON had some immune functional benefit transiently. Multiple surface markers changed only at 14 d, and not 28 d of DON exposure, suggesting that BALB/c mice adapted to DON exposure, or that this model which generally showed immunosuppression after 28 d masked DON toxicity at the later time point. (Form PFPR07) 2

152 FY07 Preliminary Final Performance Report PI: Hendrich, Suzanne USDA-ARS Agreement #: Impact: These findings support a NOAEL for dietary DON of less than 1.0 ppm, and support the need for additional studies in mice housed in groups rather than in social isolation which may have additional relevance to humans. That this study at least partly supported the hypothesis that old mice experience greater effects of DON has the impact that further studies of old mice and DON exposure are warranted. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community, agriculture and regulators have more clues as to what markers of DON exposure are most likely to be useful for human epidemiological studies of DON (T cytotoxic cells in the elderly, T helper cells and T helper chemokine receptor CCR7 in young adults, at least for shorter term exposures to DON). The scientific and regulatory communities also have additional evidence from this study that 1.0 ppm dietary DON causes changes in immune cells in the blood and spleen that need further mechanistic study to determine to what extent such changes are harmful or have benefit. Overall, this work provides important additional support for regulations to limit DON exposure to less than 1.0 ppm in the total diet. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-reviewed publications Wu X, Kohut M, Cunnick J, Hendrich S. Sex differences in apparent adaptation to immunotoxicity of deoxynivalenol. Food Chem Toxicol, submitted May Presentations Hendrich S, Wu X, Cunnick JE, Kohut M. (2008). Age and Sex differences in apparent adaptation to immunotoxicity of deoxynivalenol in BALB/c mice. Midwest AOAC meeting, Bozeman, MT, June 10, Wu X, Kohut M, Cunnick J, Hendrich S (2008) Sex differences in apparent adaptation to immunotoxicity of deoxynivalenol. Society of Toxicology annual meeting March 17, 2008, Seattle, WA. (Form PFPR07) 3

153 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Donald Hershman Institution: University of Kentucky Address: Department of Plant Pathology P.O. Box Hopkinsville St. Princton, KY Phone: ext 215 Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Uniform Trial for Integrated Control in FHB: Kentucky. Title: FY07 ARS Award Amount: $ 10,346 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Uniform Trial for Integrated Control in FHB. $10,346 Total Award Amount $ 10,346 06/24/08 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

154 PI: Hershman, Donald USDA-ARS Agreement #: Project 1: Uniform Trial for Integrated Control in FHB. 1. What major problem or issue is being resolved and how are you resolving it? Neither fungicide use nor genetic resistance to FHB can provide acceptable control of FHB symptom expression and DON accumulation when conditions favor disease development. However, when used together in an integrated program, applying fungicides to wheat varieties with FHB resistance may provide acceptable control. The work carried out in KY is being done as part of a National Uniform Integrated Control project. In Kentucky, two tests were conducted in Lexington, KY; one test was carried out in a field planted to corn the previous year and the other soybean. Both fields were planted following no-till methods. Treatments included three wheat varieties with differing levels of FHB resistance, and treated or not treated with the fungicide Prosaro at early anthesis. Plots were not inoculated or irrigated to simulate natural disease/production conditions. Disease ratings, yield, test weight, % visually scabby kernals and ppm DON data were collected. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: One test was successfully carried out and the other was accidently mowed down by the farm crew. However, this test was successfully carried out during 2008 (albeit not part of USWBSI funding). Data from the 2007 test was sent to Pierce Paul at The Ohio State University for inclusion in the Uniform Integrated Trials summary. The same will be done for the 2008 data. There was less than 1% FHB incidence in both years and very little DON accumulation (DON results from the 2008 test are pending). As a result of low disease pressure, no treatment differences were apparent for either FHB or DON. Impact: Due to low disease pressure, the Kentucky Integrated Trial reaffirmed the overwhelming importance of environmental conditions as the factor that overrides all others in determining whether or not FHB/DON will be significant. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Nothing new has been learned due to the lack of disease and no treatment effects. (Form FPR07) 2

155 PI: Hershman, Donald USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Paul, P., L Madden, M. McMullen, D. Hershman, L. Sweets, S. Wegulo, W. Bockus, S. Halley, and K. Ruden An integrated approach to managing FHB and DON in wheat: Uniform Trials Pages IN: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, MO; Dec 2-4, Hershman, D Uniform integrated FHB management trials. A presentation to stakeholders during the May 15, 2007 University of Kentucky Wheat Science Field Day, Princeton, KY. (Form FPR07) 3

156 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Charla Hollingsworth Institution: University of Minnesota Address: Department of Plant Pathology 2900 University Ave. Crookston, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 34,146 Investigating Fungicide and Application Strategies for Increased FHB Control. USWBSI Individual Project(s) USWBSI Research Area * CBCC ARS Adjusted Award Project Title Amount Investigating Ways to Improve Current Management Strategies for $34,146 FHB on Wheat and Barley. Total Award Amount $ 34,146 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

157 PI: Hollingsworth, Charla USDA-ARS Agreement #: Project 1: Investigating Ways to Improve Current Management Strategies for FHB on Wheat and Barley. 1. What major problem or issue is being resolved and how are you resolving it? Preliminary research on spring wheat conducted at several locations in the Red River Valley indicated that a flag leaf application of pyraclostrobin (Headline) significantly increased deoxynivalenol (DON) content of grain over that of the no fungicide control treatment. During five of seven test years, grain of at least one wheat cultivar contained significantly increased levels of DON from the pyraclostrobin treatment compared to the no fungicide control. Chemical company sales personnel and others routinely recommend pyraclostrobin application up to the heading growth stage (wheat) which is allowed by product registration. Recently, much confusion among plant pathologists and others has arisen do to an aggressive Plant Health marketing campaign which promotes strobilurin active ingredient application on crops (wheat, barley, corn, soybean, sugar beet, etc.) to obtain stay-green physiological effects. This stress management effect is being touted by fungicide application promoters, rather than the more traditional recommendation of disease development. Producers are applying strobilurin products more and more to reduce plant stress and increase yields irrespective of disease pressures. Increased use of this chemistry on wheat and barley is worrisome, not only because overuse will hasten pathogen resistance development, but because strobilurins are associated in the U.S. and Europe with increased DON levels in grain. This project was conducted to obtain data as described below. 1) The strobilurin chemistry offers excellent control of leaf disease on wheat and barley. It is critical, then, to determine the optimum application timing to support disease management and grain quality. We currently have no grain quality response data (DON content) from fungicide application timings prior to spike emergence. The cereal community must currently rely on fungicide marketing and manufacturing data coming from pesticide companies for best management practices concerning this issue. This research project represents the first important step in addressing this issue and in determining the latest crop growth stage that chemistries can safely be applied. 2) A second objective was to establish if wheat and barley cultivars/entries accumulated DON in grain predictably when compared across resistance levels. Specifically, we wanted to know if strobilurin application on susceptible versus resistant cultivars to Fusarium head blight (FHB) resulted in comparable DON levels. Preliminary studies indicated that wheat cultivars susceptible to FHB had increased DON, whereas cultivars with resistance were affected much less. 3) Finally, weather data (hourly air temperature, humidity, etc.) were collected near the test site to correlate environmental conditions to increased DON accumulation. Our objective was to identify those environmental parameters that increased the risk for elevated DON content. Recent list serve communications among cereal pathologists and others indicate an increasing level of frustration regarding industry marketing campaigns aimed at producers to apply strobilurin fungicides on crops in the absence of disease. As of today, 16 June, I have been made (Form FPR07) 2

158 PI: Hollingsworth, Charla USDA-ARS Agreement #: aware of a company using data produced by my program in such a manner. We currently do not have the data to response to this issue. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: This research project depended on natural infection occurring in the field for disease development. This is always risky, but one of the objectives of the project was to identify environmental conditions that promote increased DON accumulation following fungicide application. Unfortunately, the environment did not promote disease development during 2007 even though the tests were planted into a commercial field location that had been planted to corn during 2006 and Grain content of DON was slight, ranging from 0.48 ppm for wheat (Reeder, Quilt applied between Feekes 10 to 10.4) and 0.49 ppm for barley (Tradition, Stratego applied at Feekes 10.5) to 0.04 and 0.07 ppm, respectively. However, three fungicide treatments in wheat and one in barley resulted in significantly greater DON levels compared with nontreated controls. The strobilurin chemistry was present in each instance. Wheat and barley cultivars with susceptibility to FHB showed increased DON levels following an application of fungicide. Specifically, Reeder had increased DON after an application of Quilt (azoxystrobin and propiconazole) or Headline (pyraclostrobin) at the Feekes 2 or Feekes growth stages in wheat. In barley, grain of Tradition had elevated DON levels following a Feekes growth stage application of Quilt, compared with the nontreated control. These results are unexpected since two of the three treatments were applied at approximately the five leaf stage of wheat. This outcome causes us to question the value of using strobilurin chemistries on FHB-susceptible cultivars during early growth stages as well. Additional study is needed to determine if these results are repeatable. This DON effect was also present in barley, which to this PI s knowledge, has not been reported previously. Impact: This is the first time that systematic and comparative fungicide research across modes of action has been conducted in the US on wheat and barley in a natural infection system to determine whether increased DON accumulation is occurring from registered fungicide products. It is an important first step in identifying interactions between crop, fungicide, and weather that promote increased DON levels. This information is critical for developing disease management recommendations that support a safe and abundant cereal supply. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: (Form FPR07) 1

159 PI: Hollingsworth, Charla USDA-ARS Agreement #: This project must be repeated at least one more growing season when disease development is moderate to severe to determine its reliability and repeatability. Until this is done, due to the controversial nature of the research, these results should not be shared with producers. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Because of the inherent controversial nature of this research, these results will not be presented to growers until additional information has been collected and the PI is confident in the project s repeatability. Project 1A: Investigating Ways to Improve Current Management Strategies for FHB on Wheat and Barley. 3. What major problem or issue is being resolved and how are you resolving it? A uniform trial to support the development of an FHB integrated disease management strategy on wheat was discussed and organized in Bloomington, MN during This PI participated in that discussion. Researchers participating in this trial collected data on residue (type and amount present), variety response by disease resistance level, and fungicide application during 2007, during the first year of the uniform test. In Minnesota, the trial was placed at two Red River Valley locations and neither site was inoculated nor misted. 4. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: These data will contribute to understanding which disease management strategies are most important for wheat producers in the Red River Valley of Minnesota as well as in the larger spring wheat production region. We report traditional harvest data and disease development information to growers, as well as incorporate a treatment cost analysis so producers can determine whether one or more applications of fungicide result in a positive or negative revenue outcome for their operation. During 2007, all 13 varieties responded well to the growing environment at both test locations, producing excellent yields of high quality grain. Unexpectedly, fungicide application significantly increased net returns compared with the no fungicide treatment for many varieties even during a year of relatively low disease pressure. The treatment that returned the most revenue across varieties was a triple application of fungicide (seed treatment, Feekes 2, and Feekes 10.51) Impact: (Form FPR07) 2

160 PI: Hollingsworth, Charla USDA-ARS Agreement #: This is the first time that a number of PIs have worked together uniformly on an integrated FHB management trial to produce data across a number of wheat-producing states. This cooperation will result in data pooling and communication, supporting collection of a greater number of data during a shorter period of time. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: As our knowledge of additive or multiplicative effects grows concerning integrative disease management, we will be able to make more precise recommendations to growers as to what they can expect to gain, or lose, if certain disease management components are not implemented such as (1) rotation, (2) growing resistant varieties to FHB, or (3) applying fungicide. Additionally, if a correlation between strategy, weather and production returns exists, it is likely that we will begin to understand disease management outcomes as never before. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Research Proceedings and Abstracts Hollingsworth, C. R., Motteberg, C. D., Holen, D. L., and Atkinson, L. M Understanding practical outcomes from implementing FHB management strategies on spring wheat. Pages In: Proceedings National Fusarium head blight Forum. S. Canty, A. Clark, D. Ellis, and D. Van Sanford eds. 2-4 Dec Kansas City, MO. Michigan State Univ. East Lansing, MI. Extension Publications Hollingsworth, C. R Red River Valley on-farm disease management trials. Pages In: 2007 Wheat Research Review. MN Wheat Research & Promotion Council. Red Lake Falls, MN. Hollingsworth, C., Motteberg, C., Holen, D., Atkinson, L Red River Valley on-farm disease management trials. Pages In: On-Farm Cropping Trials Northwest and West Central Minnesota, Jan Hollingsworth, C., Motteberg, C., and Holen, D Today s showers bring tomorrow s (sick) flowers. In: Cropping Issues in Northwest Minnesota. Vol. 4 Issue 5. Producer meetings/workshops/symposia 13 Dec Prairie Grains Conference. On-farm disease management trial results. Grand Forks, ND: about 200 attendees. (Form FPR07) 3

161 PI: Hollingsworth, Charla USDA-ARS Agreement #: Jan Ag Professional Updates. Understanding economic outcomes from implementing disease management strategies on wheat. Crookston, MN. 60 attendees. Jan Small Grains Update Meetings. Fungicide approach to disease management and an outlook for fungicide products in Morris, Breckenridge, Moorhead, Crookston, Ada, Hallock, Argyle, Roseau, and St. Hilaire. 755 attendees total. Industry sponsored workshops/training session Nov Cereal/Pulse Crop Fungicide Conference. Response of wheat to disease management ~2007. Fargo, ND: 35 attendees. Field days 16 July Wheat disease update; identification, management review, and research summary. On-farm wheat disease management and variety trial field days, Perley and Fergus Falls: about 110 attendees total. 11 July Wheat disease update, identification, and research demonstration. Northwest Research and Outreach Center Field Day. Crookston: about 225 total attendees. 3 sessions 10 July Wheat disease update, identification, management review, and research summary. On-farm wheat variety and disease management trial field days, Hallock and Strathcona: about 90 attendees total. 9 July Wheat disease update, identification, management review, and research summary. On-farm wheat variety and disease management trial field day. Oklee: about 45 attendees. Feature articles in trade journals - mass media Hollingsworth, C., Motteberg, C., Holen, D., and Atkinson, L., On-farm disease management trials of spring wheat in the Red River Valley. Prairie Grains 92: (Form FPR07) 4

162 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Richard Horsley Institution: North Dakota State University Address: Department of Plant Sciences 166 Loftsgrad Hall Box 5051 Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement An Integrated Approach for Developing Scab Resistant Barley Title: FY07 ARS Award Amount: $ 227,369 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGG Determining the Genetic Basis of FHB Resistance and Low DON in Shenmai 3 Barley. $28,293 IIR Evaluation of Barley for FHB Resistance in China. $ 33,179 VDUN Development of Scab Resistant Six-Rowed Barley Varieties for North Dakota. $ 121,951 VDUN Development of Scab Resistant Two-Rowed Barley Varieties for North Dakota. $ 43,946 Total Award Amount $ 227,369 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07) 0

163 PI: Horsley, Richard USDA-ARS Agreement #: Project 1: Determining the Genetic Basis of FHB Resistance and Low DON in Shenmai 3 Barley. 1. What major problem or issue is being resolved and how are you resolving it? Some of the best sources of FHB resistance and low DON accumulation in barley are the Chinese accessions CIho 4196 and Zhedar 2. These accessions are extremely tall and late maturing due to a linkage block of genes found in the centromeric region of chromosome 2H. The cultivar Shenmai 3 is a Chinese cultivar with FHB resistance, low DON accumulation, early maturity, and acceptable plant height. This cultivar represents an important discovery in the search for lines to be used as sources of FHB resistance by barley breeders. The overall goal of this project is to identify molecular markers associated with FHB resistance in Shenmai 3 that can be used for marker assisted selection and to determine the interaction of genes in this cultivar controlling FHB resistance, DON accumulation, maturity and plant height. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: DArT data were collected on the mapping population. The population and parents also were grown in FHB nurseries at Langdon and Osnabrock, ND. Data on percent FHB severity, DON accumulation, plant height, and heading date were collected. Preliminary data suggest that QTL for FHB severity and DON accumulation are located in a region of chromosome 2H where QTL for these traits had been identified in Zhedar 2, CIho 4196, and Chevron. Impact: A linkage block that contains QTL for reduced FHB severity and DON accumulation, along with acceptable maturity and plant height has been identified. Because the linkage block does not contain loci for late maturity and tall plants, breeders should be able to develop varieties resistant to FHB severity and DON accumulation with acceptable maturity and plant height more efficiently. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: A source of FHB resistance and low DON accumulation with the 2H QTL for these traits that are positively linked to acceptable maturity and plant height. (Form FPR07) 2

164 PI: Horsley, Richard USDA-ARS Agreement #: Project 2: Evaluation of Barley for FHB Resistance in China. 1. What major problem or issue is being resolved and how are you resolving it? A screening nursery for FHB resistance in China has been used since An important feature of the nursery is that materials from all Midwest barley breeding programs and the ICARDA/CIMMYT barley breeding program are grown in a common environment where the only causal organism causing head blight is Fusarium graminearum. Information collected on plants grown in the nursery is shared among all collaborators in the nursery, and improved parents are identified and also shared. The nursery is located at Zhejiang University and is overseen by Professor Bingxin Zhang, a professor in the Department of Plant Protection. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: During the 11 years of growing the nursery in China, it has been used for screening accessions from various germplasm collections for new sources of FHB resistance, genotyping mapping populations for FHB resistance and DON accumulation, and screening breeding lines for FHB resistance. Over the years, new sources of resistance have been identified and over 10 different mapping populations have been screened. Accessions screened in 2007 were from the USDA-ARS-NSGC and the Vavilov Institute in Russia. Accessions, including Hordeum spontaneum, were identified with putative resistance. Advanced breeding lines from the University of Minnesota (U of M), North Dakota State University (NDSU), ICARDA, and Busch Agricultural Resources, Inc. were identified with improved FHB resistance. FHB severity data on mapping populations developed at the U of M and NDSU were collected and used for QTL mapping and marker assisted selection validation. Impact: Lines with FHB severity were identified in all breeding programs with better FHB resistance than Robust. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Multiple breeding lines and potentially wild barley accessions with improved FHB resistance as compared to Robust. (Form FPR07) 3

165 PI: Horsley, Richard USDA-ARS Agreement #: Project 3: Development of Scab Resistant Six-Rowed Barley Varieties for North Dakota. 1. What major problem or issue is being resolved and how are you resolving it? Since the inception of this project, a main objective has been to transfer FHB resistance and low DON accumulation from unadapted resistant accessions to our elite Midwest six-rowed malting barley germplasm. A limitation in this effort has been a linkage block near the centromeric region of chromosome 2H that is found in most of the resistant accessions. The linkage block includes loci controlling FHB resistance, DON accumulation, heading date, plant height, and maturity. A strategy we are using to overcome the negative linkages in chromosome 2H is to control plant height and maturity using genes from outside the critical region. For example, parents containing the sdw1 (semidwarf 1) gene in chromosome 3H were crossed to tall FHB resistant lines as a means for reducing plant height in FHB resistant progeny. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The breeding line ND20448 was found satisfactory in its second year of Pilot Scale evaluation by the American Malting Barley Association (AMBA) and was approved for advancement to Plant Scale evaluation. Approximately 600 acres of ND20448 will be grown in North Dakota during the 2008 growing season to provide grain for Plant Scale malting and brewing evaluations. Impact: A breeding line with increased resistances to FHB and DON accumulation that appears to have acceptable malting quality and agronomic performance. The line is being used by multiple breeding programs as a parent in their crossing blocks. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: ND20448 is the first lines from the NDSU six-rowed barley breeding program to reach Plant Scale evaluation by the AMBA. (Form FPR07) 4

166 PI: Horsley, Richard USDA-ARS Agreement #: Project 4: Development of Scab Resistant Two-Rowed Barley Varieties for North Dakota. 1. What major problem or issue is being resolved and how are you resolving it? The research on this project focused on improved resistance to FHB, incited primarily by Fusarium graminearum, in two-rowed spring malting barley for the Upper Midwest. Malting barley growers and their consumers need cultivars in which the toxin DON does not accumulate or is very low. Many years of breeding are required to develop such cultivars; thus, intermediate goals were developed as follows: 1) to identify good selections from crosses to FHB resistant accessions, 2) to accumulate FHB resistance by crossing good lines and selecting superior lines from their progenies, and 3) to study genetic mechanisms that contribute to reduced levels of FHB severity. Accessions previously identified as partially resistant to FHB were crossed again to elite breeding lines. Their F 5 lines were evaluated in FHB screening nurseries near Hangzhou, China and Osnabrock, North Dakota. The inheritance of genes for FHB reaction, DON accumulation, maturity, and plant height were studied in segregating progenies to better understand how to make progress in developing two-rowed barley lines with improved FHB resistance. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The cultivar Pinnacle was released in June Pinnacle has slightly better FHB resistance and accumulates similar levels of DON as Conlon. Conlon accumulates about 50% less DON than Robust and is the only two-rowed cultivars from the Midwest US that is on the AMBA list of recommended malting barley cultivars. The yield potential and straw strength of Pinnacle are superior to that of Conlon. ADM malt, Rahr Malt, Miller Brewing and Boulevard Brewing have agreed to perform Plant Scale evaluation of Pinnacle in Anheuser-Busch will perform another year of Pilot Scale evaluation of Pinnacle before deciding if they will it for Plant Scale brewing evaluation. Impact: Provided growers with a barley cultivar that accumulates similar levels of DON as Conlon, but has superior yield potential and straw strength. Pinnacle is adapted for the Red River Valley region of North Dakota and Minnesota, whereas Conlon is not. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: A reduced DON accumulating cultivar with superior yield potential and wider adaptation that Conlon barley. (Form FPR07) 5

167 PI: Horsley, Richard USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and nonpeer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Abstracts Boyd, C., R. Horsley, and A. Kleinhofs Barley chromosome 2 (2H) bin 10 Fusarium head blight resistance QTL: mapping and development of isolines. p In S. Canty A. Clark, D. Ellis, and D. Van Sanford (eds.) Proc of the 2007 National Fusarium Head Blight Forum, Kansas City, MO 2-4 Dec ASAP Printing, Okemos, MI. Presentations Update on the NDSU Barley Breeding Program at the 2008 North Dakota Barley Show, Osnabrock, ND in March (Form FPR07) 6

168 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Jerry Johnson Institution: University of Georgia Address: Department of Crop and Soil Sciences Redding Bldg 1109 Experiment St. CAES Griffin Campus Griffin, GA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 32,257 Enhancement of Scab Resistant Wheat Cultivars Adapted to the Southeast. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Enhancement of Scab Resistant Wheat Cultivars Adapted to the $32,257 Southeast. Total Award Amount $ 32,257 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

169 PI: Johnson, Jerry USDA-ARS Agreement #: Project 1: Enhancement of Scab Resistant Wheat Cultivars Adapted to the Southeast. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) epidemics have occurred in Georgia and the Southeast that resulted in marketing difficulty of grain due to high DON concentrations. Presently, scab resistant cultivars with resistance to other diseases such as leaf rust and stripe rust are not available in adequate numbers. The goal is to enhance soft red winter wheat cultivars with Fusarium head blight resistance and improved yield potential for commercial use in the lower Southeastern United States. The objectives are to identify, incorporate, and pyramid resistance to FHB in elite lines and to marker-assisted selection to pyramid FHB resistance. Both native and exotic sources of scab resistance are being identified and incorporated into elite lines. Marker Assisted Selection is also being employed to accelerate the development of adapted FHB resistant cultivars by the assistance in the selections within populations containing 3BS and 5AS. Wheat breeding lines from the southern uniform scab nurseries are being evaluated for resistance and adaptation under field conditions in the Southeast. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Scab resistant cultivars that have combined resistance to leaf and stripe rust have been lacking in the Southeast. Several wheat germplasm from both native and exotic sources with FHB resistance have been transferred into elite lines that are adapted to the Southeast. Breeding lines have been identified with native scab resistance that is equal to or better than Ernie. These lines are being increase with the intention for release. MAS with SSR markers were used to accelerate the development of scab resistance from 3BS and 5AS into elite lines. Advanced lines from GA DH and GA031454DH (derivative of Sumai 3), VA02W-713 (Ning7840 / PIO2691 // Roane), INW 0411, and Truman were further selected for field evaluation. Impact: Wheat lines from diverse origin with moderate FHB resistance are available that are adapted to the Southeast. The breeding line, GA E8 (Ernie derived) has been identified with good native scab resistance and is being increased for possible release. Two other lines, GA E34 and GA E13 have been identified in the Uniform Southern FHB nurseries with moderate Type II resistance. Both native and exotic sources of scab resistance are also being incorporated into adapted lines with good agronomic performance. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: (Form FPR07) 2

170 PI: Johnson, Jerry USDA-ARS Agreement #: Cultivars and elite lines with moderate resistance to Fusarium head blight and adapted to the southeast will be available for producers. Millers should be able to obtain grain with acceptable quality. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Jerry Johnson, Zhenbang Chen, James Buck, and Mingli Wang Introduction of Fusarium Head Blight (FHB) Resistance into Soft Red Winter Wheat AGS2000. National FHB Forum, Kansas City, KS J.W. Johnson, L. Miranda, and Z. Chen Wheat Coordinated Agricultural Project (CAP). Small Grain and Soybean Expo, Statesboro, GA. (Form FPR07) 3

171 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Nancy Keller Institution: University of Wisconsin Address: Department of Plant Pathology 1630 Linden Dr. Madison, WI Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Genetic Mechanisms to Control Head Scab. Title: FY07 ARS Award Amount: $ 22,630 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount PGG Global Regulation of Fusarium Toxins. $22,630 Total Award Amount $ 22,630 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

172 PI: Keller, Nancy USDA-ARS Agreement #: Project 1: Global Regulation of Fusarium Toxins. 1. What major problem or issue is being resolved and how are you resolving it? One of the most severe mycotoxin problems in the U.S. is trichothecene contamination of small grains by Gibberella zeae (anamorph Fusarium graminearum) in a disease called scab or Fusarium head blight (FHB). Our lab is attempting to find genes and gene products important in either Gibberella zeae sporulation or toxin production. Such genes and gene products would provide needed knowledge of virulence factors in the fungus and possibly provide insight into control strategies.. Recent progress in fungal toxin clusters has suggested that these clusters, as well as spore formation, is globally regulated by higher order chromatin conformation. For this grant, we have initiated studies to identify global regulators of secondary metabolite (e.g. mycotoxin) gene clusters in Fusarium based on our findings from the genus Aspergillus. Specifically we have identified and inactivated hepa, a putative heterochromatin protein 1 involved in heterochromatin formation in F. graminearum. We will now examine our hypothesis that deletion of hepa increases toxin formation through activation of chromatin. We also have identified demethylase genes that we predict will decrease toxin formation through repression of chromatin. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Toxin genes in fungi are typically arranged in clusters and our work in Aspergillus species shows these clusters are located in facultative heterochromatic regions that when activated (euchromatin format) result in increased toxin production but when silenced (heterochromatin format), decreased toxin formation. Our initial results suggest toxin formation may also be regulated by heterochromatin in Fusarium spp, Impact: This is the first identification of F. graminearum histone remodeling genes, a topic that has become of interest in international laboratories. While it is yet too early to assess the role of these genes on fungal toxin production, we expect there will be significant effects. Several other labs have shown that chromatin remodeling proteins and interacting partners affect Fusarium mycotoxin production and spore production based on our findings in Aspergillus. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: An important contribution of this work the importance of chromatin remodeling on toxin gene cluster expression which appears to be taken up by the research community world wide. Several of the proteins we have identified can be considered for fungicide targeting. (Form FPR07) 2

173 PI: Keller, Nancy USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. From past and present SCAB funding, the Fusarium hepa work is not yet completed Palmer J, Perrin R, Keller NP (submitted) H3K9 methylation regulates growth and development in Aspergillus fumigatus. Euk Cell Tsitsigiannis D I, Keller NP (2007) Oxylipins as developmental and host-fungal communication signals. Trends in Microbiology Mar;15(3): Brodhagen M, Keller NP (2006) Signaling pathways connecting mycotoxin production and sporulation. Molecular Plant Pathology 7: McDonald T, Devi T, Shimizu K, Sim S-C, Keller NP (2004) Signaling events connecting mycotoxin biosynthesis and sporulation in Aspergillus and Fusarium spp. In New Horizon of Mycotoxicology for Assuring Food Safety, Proceedings of the International Symposium of Mycotoxicology (Editor: Takumi Yoshizawa) pp (Form FPR07) 3

174 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: James Kells Institution: Michigan State University Address: Department of Crop & Soil Sciences 286 PSSB East Lansing, MI Phone: ext. 103 Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 96,660 Conventional Breeding Information Technologies to Control FHB in Wheat and Barley. USWBSI Individual Project(s) USWBSI Research Area * EC/HQ ARS Adjusted Award Project Title Amount Hosting the Manager of the USWBSI's Networking and Facilitation $96,660 Office. Total Award Amount $ 96,660 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

175 PI: Kells, James USDA-ARS Agreement #: Project 1: Hosting the Manager of the USWBSI's Networking and Facilitation Office. 1. What major problem or issue is being resolved and how are you resolving it? Scab affects the industries and people involved in virtually every stage of the production, processing, and distribution systems of five market classes of wheat and barley across the U.S. The Networking and Facilitation Office (NFO) has been in existence since The goal of the NFO is to minimize the barriers to the success of the U.S. Wheat & Barley Scab Initiative arising from the administrative burdens and communication challenges. In FY07, there were 85 researchers working in 27 institutions (including ARS and CIMMYT) across 25 states and Mexico. The NFO is addressing this problem by 1) helping scientists and all interested parties, who in the past may have problems communicating with each other, to learn from each other by facilitating communications and communication system development; 2) identifying and implementing an internet-based communication and collaboration mechanisms; and 3) acting as a center of accountability and a rapid clearing house of scab-related information. The NFO provides administrative support to the various committees, as well as being responsible for the organization of the Initiative s annual scientific conference. Finally, the NFO facilitates the annual resolution of a comprehensive national research plan and budget, and represents the Initiative s only fixed asset. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment (1): During FY07, the USWBSI began implementation of its newly developed Action Plan starting with the creation of five coordinated Projects: Commodity-based Coordinated Projects: o Barley Coordinated Project o Hard Winter Wheat Coordinated Variety Development and Host Resistant Coordinated Projects o Spring Wheat Parents (included Durum projects) o Northern Winter Wheat o Southern Winter Wheat Impact (1): By coordinating research efforts within these specific categories, the expectation is that it will lead to greater collaboration and communication, improved focusing of research efforts and more efficient use of resources. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (1) Several of these coordinated projects include their stakeholders (growers, producers, endusers) in the development of the research plan for their commodity or region. Researchers have more involvement in determining the research priorities for their coordinated project. (Form FPR07) 2

176 PI: Kells, James USDA-ARS Agreement #: Accomplishment (2): One of the NFO s most important accomplishments was the successful facilitation of the development of the USWBSI s FY08 Research Plan and Budget, which was approved by USDA-ARS in February. See attached table (1), for a summary of meetings, conference calls, etc. facilitated by the NFO that lead to the FY08 Research Plan and Budget. Impact (2): One-hundred and twenty-four research projects across five research areas, and the USWBSI Headquarters, received funding in the amount of $4,956,797 to continue the fight against FHB. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?:(2) There are 83 scientists in 25 states and Mexico continuing to work on developing as quickly as possible effective control measures that minimize the threat of Fusarium Head Blight (Scab) to the producers, processors, and consumers of wheat and barley. Accomplishment (3): In December of 2007, the NFO working with the Forum Organizing Committee facilitated the USWBSI s annual scientific Forum, which was held in Kansas City, Missouri. Over 200 crop scientists (including research associates, research technicians and graduate students), growers, and industry representatives from across the nation and around the world, as well as area leaders of the U.S. wheat and barley industry, attended the 2007 National FHB Forum. Research results and findings were presented to the forum participants in the form of invited talks (22), and poster (102) presentations accompanied by abstracts or papers published in the Forum proceedings. One afternoon was devoted to group discussions designed around the USWBSI s five research areas and three commodity-based coordinated projects. The focus of the group discussions was the implementation of the USWBSI s Action Plan and communicating results in the real world. There were also mini breakout sessions covering the following topics: Tebuconazole EPA Emergency Exemption: Recent Developments and 2008 Application Positioning; Genotyping Centers Breeders met with scientists from the genotyping labs. FHB Disease Forecasting Models; and Technicians and graduate students: an informal exchange of ideas and protocols for FHB research. Impact (3): The annual Forum is the primary mechanism whereby USWBSI scientists and stakeholders come together to discuss both the progress being made, as well as problems that interfere, with the successful combat against this devastating disease. (Form FPR07) 3

177 PI: Kells, James USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (3) The clientele has a greater understanding of what research is being conducted under the USWBSI; as well what progress is being made in the following areas (titles of Forum poster sessions): Host Plant Resistance and Variety Development Gene Discovery and Engineering Resistance Pathogen Biology and Genetics Food Safety, Toxicology and Utilization of Mycotoxin-contaminated Grain FHB Management Table 1. Summary of Meetings, Conference Calls and Workshops facilitated by the Networking & Facilitation Office (NFO) during FY07. # of Meetings/ Committee Conference Call (CC) Description/Purpose of Meeting Steering Committee (SC) 2 Meetings : Semi-Annual Meeting : Semi-Annual Meeting Review FY08 Research Plan and Executive Committee (EC) 4 Meetings Budget Meetings: Follow-up to SC s meeting; met with review panels to discuss their recommendations; follow-up of Annual Forum and USWBSI s recommendation for FY08 Research Plan and Budget. USWBSI SC-EC Sub Committees Coordinated Project Committees (CPC) /Review Panels (RP) RA Workshops/ Planning Meetings 2 CC 1 Meeting 7 CC CPCs 8 CC RP 11 CC 4 Workshops/ Meetings 7 CC Conference Calls: Discuss USWBSI Business Annul Research Plan and Budget; setting of Working Caps; agenda for semi-annual spring SC meeting; and program for Annual Forum Communication sub-committee Meeting, Fargo, ND Communications sub-committee confernce call Purpose of meetings: Enhancing the USWBSI s communication activities EC sub-committee interview with Co-Chair candidates 2-08 PBG sub-committee - discussed future direction of PBG research area. Coordinated Project Committees (CPC) Discuss final FY08 coordinated project pre-proposal. Review Panels - Discuss FY08 Funding Recommendation 6-07 Barley Coordinated Project Planning Meeting, St. Paul, MN 6-07 DON Labs discuss protocols for sampling for DON VDHR Northern Winter Wheat Coordinated Project Conference Call 7-07 VDHR-Spring Wheat Coordinated Project Conference Call 8-07 FHB Management Biocontrol Planning Meeting, San Diego, CA 3-08 VDHR Soft Winter Wheat Coordinated Project Planning Meeting, Wooster, OH 3-08 PBG RAC conducted three separate conference calls with MGMT, GDER and VDHR research scientists/committee members FSTU discuss coordination of DON testing labs VDHR-Spring Wheat Coordinated Project Plannning Meeting, Fargo, ND (Form FPR07) 3

178 PI: Kells, James USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Forum Proceedings: Canty, S., A. Clark, D. Ellis and D. Van Sanford (Eds.) Proceedings from the 2007 National Fusarium Head Blight Forum, Kansas City, MO; December 2-4. University of Kentucky, Lexington, KY. 252pp. Proceedings are available in PDF format on the USWBSI s Website ( Invited Talks: Van Sanford, D. A The USWBSI: Status Report Wheat Quality Council Annual Meeting. Kansas City, MO. February 19-21, Newsletters: Lilleboe, D., S. Canty, and D. Van Sanford. Spring Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 8, Issue 1. Lilleboe, D., S. Canty, and D. Van Sanford. Fall Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 7, Issue 2. Press Releases: News Release: Scab Incidence Varies Across Midwestern States News Release: Early Southern Scab Reports Mixed News Release: USWBSI Announces new Co-Chair News Release: Scab Disease Generally Low across U.S. in Press Release: USWBSI Mourns Loss of Co-Chair Tom Anderson. All Press Releases are available in PDF format on the USWBSI s Website ( (Form FPR07) 4

179 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Shahryar Kianian Institution: North Dakota State University Address: Department of Plant Sciences 470G Loftsgard Hall Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 39,024 Development of Markers Linked to FHB Resistance in Durum and Hexaploid Wheat. USWBSI Individual Project(s) USWBSI Research Area * HGG ARS Adjusted Award Project Title Amount Pedigree Based Association Analysis of New FHB Resistance $39,024 Sources in Wheat. Total Award Amount $ 39,024 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

180 PI: Kianian, Shahryar USDA-ARS Agreement #: Project 1: Pedigree Based Association Analysis of New FHB Resistance Sources in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Sources of FHB resistance are constantly being sought to expand the number of available genes to pyramid in a single variety. However, current methods rely on developing either recombinant inbred line (RIL) or double haploid (DH) mapping populations from resistant by susceptible crosses, phenotyping in multiple replicated trials, mapping and QTL analysis. This is then followed by the transfer of desirable regions with linked markers into acceptable cultivars. This process is tedious and slows the process of cultivar development. In this study we are exploring an alternative strategy that will allow us to not only quickly map the region of interest from multiple sources but do this in breeding populations under development. Our strategy is intended to reduce the time required for QTL identification and marker assisted selection significantly. The North Dakota Durum Wheat breeding program has identified five tetraploid wheat sources of resistance from Tunisia, which were selected among a large number of lines evaluated over 55 repeated FHB trials. Since their identification these lines have been extensively used in the breeding program to derive resistant breeding lines. We plan to use this collection of backcross derived advanced resistant lines, susceptible sibs, and parental lines to identify markers that are associated with these novel sources of resistance. Again the beauty of this analysis is the integration of derived information directly into the breeding programs. Due to significant reduction in funding by USWBSI our objectives for this time period were: 1) evaluate the parental (resistant donor and susceptible recipient) with PCR based markers evenly spaced (10 cm intervals) throughout the wheat genome; 3) to a very limited extend analyze the derived lines (all lines in a given pedigree) for polymorphic markers; and 3) screen all advanced lines with minimal FHB screening data in the greenhouse and field nursery. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: In this study we used 184 BC1F6 and 189 BC1F7 lines derived from crossing of Tun7, Tun18, Tun34, Tun36 with durum cultivars Ben, Maier, Lebsock and Mountrail for association studies. As Tunisian lines pedigree shows no relation to Chinese genotypes, they probably carry different genes or alleles for resistance to FHB. We checked all the parents and RILs in the greenhouse in three seasons for type II resistance to FHB by single floret injection inoculation method. The data showed that the Tunisian lines have different amount of resistance varying from 18% to 10% infection rate through the spikes. The data also showed Maier may have some minor resistance genes because it showed a moderate resistance in our greenhouse study and the crosses between Maier and different Tunisians had more transgressive resistant progenies compare with the other crosses. To accelerate the identification of markers associated to FHB resistance, we initially screened the parents. The amounts of recombination in wheat chromosome arms are low so we picked SSR markers per chromosome which were roughly 10cm apart and cover the whole genome. Among the 179 SSR markers that we applied on the parents about 45% showed polymorphism for at least two parents and about 8 % showed polymorphism between (Form FPR07) 2

181 PI: Kianian, Shahryar USDA-ARS Agreement #: the whole set of Tunisian lines and susceptible cultivars. The most polymorphism was found on chromosomes 5A and 3B and the least on chromosome 6A. About 22 SSR markers that had been mentioned in different articles to be linked to FHB resistant were also applied to the parents. Among them barc117 and gwm129 from chromosome 5A showed the same pattern in Tunisian lines but not the susceptible lines. Impact: This is the first time an analysis of FHB resistance in tetraploid wheat lines of Tunisian decent is being performed. This is the first step toward identification and pyramiding of FHB resistance loci from various sources. Also this is the first time a study of this type, utilizing pedigree information to identify and incorporate new genes for resistance in wheat cultivars, is being performed. Again the beauty of this analysis is the integration of derived information directly into the breeding programs. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: New germplasm, new markers associated with novel FHB resistance regions, and new methodology to directly characterize and incorporate traits into breeding lines. These all will have a major impact on how we perform QTL analysis in the future and incorporate the results into an applied breeding program. (Form FPR07) 3

182 PI: Kianian, Shahryar USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Ghavami F., M. Huhn, E. Elias, and S.F. Kianian Tunisian durum wheat as new sources of resistance to Fusarium head blight. Proceedings of the 2007 National Fusarium Head Blight Forum. Kansas City, MO. Pages Poster #78. Ghavami F., and S. F. Kianian Pedigree based association analysis of new FHB resistance sources in durum wheat. 5 th Annual Northern Plains Biological Symposium. Fargo, ND. (Form FPR07) 4

183 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: William Kirk Institution: Michigan State University Address: Department of Botany & Plant Pathology 35 Plant Biology Building East Lansing, MI Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 13,114 Field Evaluations of Chemical Control for Fusarium Head Blight in Michigan. USWBSI Individual Project(s) USWBSI Research Area * CBCC ARS Adjusted Award Project Title Amount Field Evaluations of Chemical Control for Fusarium Head Blight in $13,114 Michigan. Total Award Amount $ 13,114 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

184 PI: Kirk, William USDA-ARS Agreement #: Project 1: Field Evaluations of Chemical Control for Fusarium Head Blight in Michigan. 1. What major problem or issue is being resolved and how are you resolving it? Nationally, Michigan ranks 15th in production of wheat with 590,000 harvested acres with a crop value of $122.6 million. The majority of plantings are in soft white and soft red winter wheat cultivars. Fusarium head blight (FHB) epidemics continue to be responsible for enormous yield and quality losses of wheat and barley (McMullen, et al., 1997) resulting in financial damage to producers, and risk to the safety of food supplies as a result of the production of mycotoxins by the causal organism. Severe widespread epidemics in the US in 1993, 1996, and 1997, and recent localized epidemics, have necessitated finding fungicides that are effective against FHB. The complex nature of the host/pathogen interaction has made control of head scab challenging and immediate options are needed to keep this disease from causing severe economic loss to farmers. Much fungicide evaluation work for FHB remains to be accomplished in Michigan. Trial results for the past several years have been inconclusive. Folicur 3.6 had been registered under a section 18 emergency exemption in Michigan as a single application (4 fl oz./acre) applied up until flowering, and Tilt and Propimax EC (Propiconazole) are currently labeled under a section 24C special local needs exemption in Michigan for suppression of FHB, products that may prove more efficacious need to undergo successful evaluation under Michigan conditions to provide additional options for producers. The research conducted in MI tested new products that have been and others that may be registered in the near future; provided supporting documentation for the continuation of Section 18 registration or for the registration of new products; and provided additional testing sites in important wheat producing areas in Michigan. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Trials on FHB and DON conducted in MI in 2006/07 indicated that effective fungicides were available for use. The data from these trials were used to support the registration of new fungicides such as Proline (prothioconazole) and Prosaro (tebuconazole + prothioconazole) Impact: Wheat growers have effective fungicides available under Section 3 registration for control of FHB and reduction of DON. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Increased food and crop security. (Form FPR07) 2

185 PI: Kirk, William USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Brown-Rytlewski, D., W.W. Kirk, R.L. Schafer and D. Berry Evaluation of fungicides for control of Fusarium head scab of winter wheat at Williamston, MI, :CF Brown-Rytlewski, D., W.W. Kirk, R.L. Schafer and D. Berry Evaluation of fungicides for control of Fusarium head scab of winter wheat at Sandusky, MI, :CF Brown-Rytlewski, D., W.W. Kirk, R.L. Schafer and D. Berry Evaluation of fungicides for control of Fusarium head scab of winter wheat at East Lansing, MI, :CF Brown-Rytlewski, D., W.W. Kirk, R.L. Schafer and D. Berry Evaluation of fungicides for control of Fusarium head scab of winter wheat at Saginaw, MI, :CF019. (Form FPR07) 3

186 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: H. Corby Kistler Institution: USDA-ARS Address: Cereal Disease Laboratory University of Minnesota 1551 Lindig Street St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Fungal Genes in DON Biosynthesis. Title: FY07 ARS Award Amount: $ 34,964 USWBSI Individual Project(s) USWBSI Research Area * PGG ARS Adjusted Award Project Title Amount Identifying Fungal Genes and Plant Factors Involved in DON $34,964 Production. Total Award Amount $ 34,964 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

187 PI: Kistler, H. Corby USDA-ARS Agreement #: NA Project 1: Identifying Fungal Genes and Plant Factors Involved in DON Production. 1. What major problem or issue is being resolved and how are you resolving it? The fungus Fusarium graminearum causes extensive losses on wheat and barley crops world-wide and contaminates harvested grain with a compound known as DON, whose levels in grain are strictly regulated. In addition to factors reducing the impact of Fusarium head blight, novel methods for reduction of DON accumulation in grain are desirable. Currently, little is known about the pathogen factors that influence the accumulation of DON in plants. This study directly addresses the issue by identifying genes responsive the fungal regulatory pathways known to influence DON accumulation in both wheat and barley. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: To elucidate their genome-wide impact on gene regulation, Tri6 and Tri10 deletion mutants were constructed in F. graminearum and used for microarray analyses with RNA isolated from infected wheat heads with the wildtype as the control. While only a total of 37 genes had expression levels altered two-fold in the Δtri10 mutant, 208 genes were altered twofold the Δtri6 mutant including transcript levels for nearly all known Tri genes. Among those also reduced were genes coding for enzymes in the isoprenoid biosynthetic pathway from acetyl CoA to farnesyl pyrophosphate, the latter being the immediate molecular precursor to all trichothecenes. Impact: This study demonstrates that the FHB fungus is remarkably adapted for producing DON, not only by precisely regulating the genes unique to toxin synthesis but also by modifying gene expression in basic house-keeping functions of the cell to promote toxin accumulation. Fungal cells thus become finely tuned toxin factories distinct from their non-toxin producing cousin species. This study also shows that alterations in the cellular factory s toxin assembly line can drastically reduce production of DON and the amount of disease caused by the fungus. These alterations have the potential to be used by FHB researchers as potential targets for designing control strategies aimed at reducing toxin concentrations in grain. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The published data on F. graminearum genes regulated by Tri6 and Tri10, as well as the accompanying microarray data has been submitted to the Plant Expression Database (plexdb.org), and represent an important transfer of technology because FHB researchers now have access to the entire suite of genes regulated by these trichothecene biosynthetic cluster genes. (Form FPR07) 2

188 PI: Kistler, H. Corby USDA-ARS Agreement #: NA Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Seong, K., Pasquali, M., Song, J., Hilburn, K., McCormick, S., Dong, Y., Xu, J.-R. and Kistler, H.C. 200x. Global gene regulation by Fusarium transcription factors Tri6 and Tri10 reveals adaptations for toxin biosynthesis. In revision for Mol. Microbiol. Pasquali, M., Lysoe, E., Seong, K.-Y., Xu, J.-R., Ma, L.-J., Kistler, H.C Global gene expression during plant infection and toxin biosynthesis in Fusarium graminearum. Journal of Plant Pathology 90 (3, Supplement): S Abstract. Pasquali, M., Seong, K.-Y., Menke, J., Lysoe, E., Hilburn, K., Xu, J.-R., and Kistler, H.C Spore development and trichothecene mutants of Fusarium graminearum. In: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, MO, 2-4 December, Page 32. Abstract. (Form FPR07) 3

189 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Andris Kleinhofs Institution: Washington State University Address: Department of Crop and Soil Science P.O. Box Johnson Hall Pullman, WA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 48,780 Saturation Mapping of the Chromosome 2(2H) Fusarium Head Blight Resistance QTL. USWBSI Individual Project(s) USWBSI Research Area * HGG ARS Adjusted Award Project Title Amount Fractional Analysis of Chromosome 2(2H) Fusarium Head Blight $48,780 Resistance QTL. Total Award Amount $ 48,780 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

190 PI: Kleinhofs, Andris USDA-ARS Agreement #: Project 1: Fractional Analysis of Chromosome 2(2H) Fusarium Head Blight Resistance QTL. 1. What major problem or issue is being resolved and how are you resolving it? Development of commercially acceptable cultivars with FHB resistance and good agronomic qualities is the goal of the barley SCAB project. One of the best FHB resistance QTL resides in the chromosome 2(2H) bin 10 region. Our contributions are focused on genetic and physical mapping of this region with the long-term goal of saturating the region with molecular markers and cloning the genes responsible for FHB resistance. To facilitate this, we have isolated recombinant lines with introgressed small chromosome 2(2H) bin 10 genomic segments in a susceptible genomic background and we have developed Bacterial Artificial Chromosome (BAC) contigs for all molecular markers mapping to this region. We have also developed 6- rowed recombinants in the resistant CI4196 genomic background. To further facilitate development of agronomically acceptable barley cultivars with FHB resistance, we have undertaken to modify the resistant line CI4196 by mutagenesis. Mutants with desirable traits such as semi-dwarf, early and 6-rowed are easily selected. These provide improved FHB resistant parent material that can be rapidly incorporated in breeding programs. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: 1. Development of BAC clone contigs for a significant portion of the FHB QTL Impact: 1. Development of BAC contigs will result in a complete physical map of the FHB QTL, development of new molecular markers for the region, sequencing of the region and identification of candidate FHB resistance genes As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: 1. Additional molecular markers targeted to the FHB resistance QTL region are needed for molecular marker assisted selection of FHB resistant cultivars. Physical contigs of the FHB QTL allow the scientific community a better understanding of the physical size of the QTL leading to improved strategies for cultivar development. Accomplishment: 2. Development of recombinants combining 6-rowed phenotype with FHB resistance Impact: 2. The 6-rowed FHB resistant phenotype is desired by the malting industry As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: 2. Breeders have access to improved FHB resistant parents for breeding cultivars Accomplishment: 3. Isolation of mutants in FHB resistant CI4196 genomic background with improved stature, earliness and 6-rowed trait Impact: 3. FHB resistant parents with improved agronomic traits available for breeders (Form FPR07) 2

191 PI: Kleinhofs, Andris USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: 3..Breeders have access to improved FHB resistant parents for breeding cultivars. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Boyd, C. N., R. Horsley and A. Kleinhofs (2007) Barley chromosome 2(2H) bin 10 Fusarium Head Blight resistance QTL: mapping and development of isolines. In: Canty, S., A. Clark, D. Ellis and D. van Sanford (Eds), Proceedings of the 2007 National Fusarium Head Blight Forum; Dec. 2-4, 2007; Kansas City, Missouri. pp (Form FPR07) 3

192 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Paul Knight Institution: Pennsylvania State University Address: Dept. of Meteorology 605 Walker Bldg. University Park, PA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 27,847 Enhanced Tools for the Deployment of Fusarium Head Blight Predication Models. USWBSI Individual Project(s) USWBSI Research Area * EEDF ARS Adjusted Award Project Title Amount Enhanced Tools for the Deployment of Fusarium Head Blight $27,847 Prediction Models. Total Award Amount $ 27,847 Principal Investigator July 2, 2008 Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

193 PI: Knight, Paul USDA-ARS Agreement #: Project 1: Enhanced Tools for the Deployment of Fusarium Head Blight Prediction Models. 1. What major problem or issue is being resolved and how are you resolving it? The issue of properly assessing the risk of FHB on a very small spatial scale by using both finer scale atmospheric data (called RTMA) and atmospheric prediction models is the primary issue. In addition, since the finer scale atmospheric data, RTMA, is interpolated/downscaled, it inherently has biases related to both the initial data and the downscaling process. Therefore, we have acquired a separate data stream from the agricultural networks (agnets) in the Plains and Great Lakes region to test two issues. The first is to compare the FHB risk tools for the agnets with the fine scale atmospheric data, RTMA, on the same interface. The second is to use historical data (2007 agnet data available for North Dakota and Michigan) to determine a bias correction factor based on differences between agnet and RTMA from last season. These bias corrected FHB predictions are available in real-time on the web interface so that the wheat growers community can determine whether the risk is the same, greater than or less than the RTMA based on the bias correction. This employs state of the art prediction techniques. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A very sophisticated interface which brings together more than 4 large data streams in realtime then presents the user with the tools needed to make decisions concerning the health of their wheat/barley crops. In addition to several plant pathology models (winter vs spring wheat and four variations of spring wheat), there are risk assessments based on just past data or adding up to 2 and half days of predicted atmospheric conditions for planning purposes. Impact: The addition of a bias correction forecast in Michigan and North Dakota this season is expected to reduce the occurrence of false positive predictions of the disease. This prototype can then be applied to the other states with agnets (whose data we are acquiring daily to determine model biases in their region). As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The clientele currently have the smallest scale of hourly data (5km)as input to the best plant pathology models using a highly sophisticated technique of prediction (bias correction) in atmospheric science. (Form FPR07) 2

194 PI: Knight, Paul USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. No publications to date. (Form FPR07) 3

195 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Frederic Kolb Institution: University of Illinois Address: Department of Crop Sciences 1102 S. Goodwin Ave. Urbana, IL Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Development of Scab Resistant Soft Red Winter Wheat Varieties. Title: FY07 ARS Award Amount: $ 100,549 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGG Mapping QTL Associated with Kernal Retention Scab Resistance in Wheat. $26,341 VDUN Development of Scab Resistant Soft Red Winter Wheat Varieties. $ 74,208 Total Award Amount $ 100,549 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

196 PI: Kolb, Frederic USDA-ARS Agreement #: Project 1: Mapping QTL Associated with Kernal Retention Scab Resistance in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Production of sound, plump kernels rather than shriveled or tombstone kernels when wheat is infected with scab is an important component of host plant scab resistance. Some lines produce sound, unshriveled kernels in spite of severe scab symptoms in the spike. In these lines the percentage of shriveled kernels produced is lower than would be expected on the basis of the level of symptoms observed in the heads. We refer to this as kernel retention. Identification of QTL conferring kernel retention resistance (from a source other than Sumai 3 derived lines) would facilitate combining scab resistance genes from different sources to produce transgressive segregants with both Type II resistance and kernel retention. The plump kernels produced must also be evaluated for DON content. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The second year of greenhouse and field data were collected for FHB symptoms (AUDPC) and percentage of Fusarium damaged kernels (FDK) on the 269 F 6:8 RILs we developed from the cross IL / Patton. Thus, phenotypic data on kernel retention were collected twice in the greenhouse, and twice in the misted, inoculated FHB evaluation nursery. The phenotypic data were combined with molecular marker data. Four severity quantitative trait loci (QTL) were identified on chromosomes 2B, 3B, 4B, and 6B, each explaining about 4% to 6% of the phenotypic variation. Three kernel damage QTL were identified on chromosomes 2B, 4B, and 6B, with 4B QTL explaining 7% and 12.3% of the phenotypic variation for kernel damage in the greenhouse and field trials, respectively. This is the second report of a kernel damage QTL in this region of chromosome 4B, indicating that this QTL may contribute to reducing FDK percentage. Impact: Although the information generated by this research will not be of direct use to wheat producers or consumers, information from this project combined with knowledge gained from other research should enhance the selection efficiency for scab resistance which will benefit producers and consumers in the long-term through the development of improved scab resistant varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The primary users of the information generated by this project will be other scientists working on scab resistance. This information on the kernel retention type of resistance was made available to the scientific community through a poster at the 2007 ASA/CSSA meeting and the 2007 Scab Forum, and a manuscript describing this research is in preparation. This information may contribute to the overall goal of understanding FHB resistance. Accomplishment: In and field seasons, to better understand the kernel retention type of resistance, we examined kernel retention in two groups of breeding lines; one group of lines that differ in kernel retention but have the same FHB index and a second group that have the same kernel retention percentages but differ in FHB index. These groups of lines include lines with diverse pedigrees and were evaluated in the mist-irrigated, (Form FPR07) 2

197 PI: Kolb, Frederic USDA-ARS Agreement #: inoculated field scab evaluation nursery. DON levels were determined on samples from both 2006 and Impact: This experiment examines the relationship among percentage of FDK, DON level and FHB field symptoms and will provide information that will aid in the development of wheat varieties with low DON levels. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Breeders will have information that will aid in the development of wheat varieties with low DON levels and low percent FDK. A manuscript describing this research is in preparation. (Form FPR07) 3

198 PI: Kolb, Frederic USDA-ARS Agreement #: Project 2: Development of Scab Resistant Soft Red Winter Wheat Varieties. 1. What major problem or issue is being resolved and how are you resolving it? The major issue is that producers need varieties with high levels of scab resistance. We are working on the development of high-yielding, well-adapted, scab resistant lines. As more lines with good scab resistance are identified we are using these parents in crosses, so that in many crosses both parents, or two parents out of three in a three-way cross, are scab resistant. We also believe that it is important to combine several types of resistance rather than rely solely on Type II resistance. We are addressing this by using the ISK index to select breeding lines with high levels of scab resistance. Development of varieties with low DON levels is also crucial; therefore, all breeding lines are evaluated each year for DON level. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Forty-eight varieties from the Illinois State Variety Trial were evaluated for FHB resistance in a FHB evaluation nursery, and data were made available to producers. Impact: In order to use FHB resistance as a criterion in variety selection producers must have as much information as possible on FHB resistance. The FHB resistance data provide very useful information to Illinois seedsmen and producers and allows them to use FHB resistance as a criterion in variety selection. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Producers and seedsmen have a three year summary of data of FHB resistance and DON level that can be used in decisions about what varieties to produce. The information on FHB resistance was made available at < Accomplishment: About 288 breeding lines in cooperative nurseries including the Uniform Northern Winter Wheat Scab Nursery, the Uniform Preliminary Northern Winter Wheat Scab Nursery, the Uniform Southern Scab Nursery, the Uniform Eastern Soft Winter Wheat Nursery, and the Adv. and Prelim. Five-State Nurseries were evaluated for FHB resistance in a misted, inoculated FHB field nursery. Lines from the Univ. of Illinois program were submitted for all of the cooperative nurseries except the Uniform Southern Scab Nursery, thus, breeding lines with FHB resistance were made available to other breeding programs for use as germplasm. Five University of Illinois lines were among the most FHB resistant lines in the 2007 PNUWWN. Impact: The data provided were useful to many different breeding programs in making decisions about which breeding lines merit further evaluation as varieties and which breeding lines will be useful as germplasm. Exchange of FHB resistant breeding lines among programs is essential and will contribute to the development of FHB resistant varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Obtaining FHB resistance data for entries in the cooperative nurseries from many environments allow wheat breeders to make better selection decisions about what lines to advance for further evaluation. Breeding lines from the University of Illinois breeding program were made available to other breeding programs for use as parents if the breeders wish to use them. (Form FPR07) 4

199 PI: Kolb, Frederic USDA-ARS Agreement #: Accomplishment: In 2007, about 415 breeding lines from the University of Illinois wheat breeding program were evaluated in the misted, inoculated scab evaluation field nursery. Scab resistant lines were evaluated for many additional traits including grain yield, milling and baking quality, standability, and resistance to other diseases. Impact:. Sustained annual selection for FHB resistance in the inoculated, misted field nursery has significant long-term impact by assuring that new varieties will be FHB resistant. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Constant selection for FHB resistance in the breeding program is essential in order to identify breeding lines with FHB resistance and also to discard FHB susceptible lines early so that resources are not wasted evaluating FHB susceptible lines. The constant selection pressure applied using evaluation in misted, inoculated nurseries is essential in reducing DON. Accomplishment: In 2007, 1793 wheat samples were sent to the lab at the University of Minnesota for DON analysis. Impact: DON evaluation is an essential component of FHB resistance evaluation because new varieties must have not just lower FHB field symptoms but also reduced DON content. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: This is information that is primarily useful to the wheat breeder, but information on low DON producing varieties can also be used by the producer in variety selection. Accomplishment: In , 236 single crosses and 202 three-way and four-way crosses were made involving FHB resistance sources. For a few of these populations MAS will be used for F 2 enrichment for the 3BS FHB resistance locus. MAS for the 3BS locus is also being used for some additional populations including a backcrossing project to introgress the 3BS locus into IL Impact: The crosses of scab resistant parents by adapted high yielding parents will provide populations that can be used for development of scab resistant varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These crosses are the source of variability that will be used for future development of scab resistant soft red winter wheat varieties. Accomplishment: Four soft red winter wheat breeding lines combining FHB resistance equal to or better than Ernie with high yield potential were released in 2007 for brand labeling and potential commercial production. Impact: The lines that enter commercial production will provide seedsmen and producers with additional FHB resistant varieties. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The availability of improved varieties with FHB resistance provides additional choices for seedsmen and producers and contributes to an overall reduction in DON and decreased susceptibility to FHB. (Form FPR07) 5

200 PI: Kolb, Frederic USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Non-refereed bulletins, reports and publications: Kolb, F.L Fusarium head scab evaluation of variety trial entries at Urbana, Illinois. Tables of data prepared for field days, variety trial report, and posted on Variety Trial website ( Kolb, F.L Development of Fusarium head blight resistant wheat varieties. Proceedings of the Illinois Crop Protection Technology Conference. Jan. 3-4, p Abstracts: Bonin, C.M., and F.L. Kolb Resistance to kernel damage caused by Fusarium head blight in an RIL population. ASA-CSSA-SSSA Annual meeting, Nov. 2-4, 2007, New Orleans, LA. Abstract included on meeting CD. Bonin, C.M., F.L. Kolb, and E.A. Brucker QTL associated with reduced kernel damage and resistance to Fusarium head blight in wheat. Proceedings of the 2007 National Fusarium Head Blight Forum. Kansas City, MO, December 2-4, 2007, page 168. Bonin, C.M., and F.L. Kolb Resistance to kernel damage caused by Fusarium head blight in an RIL population. Proceedings of the 2007 National Fusarium Head Blight Forum. Kansas City, MO, December 2-4, 2007, page 169. Perugini, L., C. Sneller, F. Kolb, D. VanSanford, C. Griffey, H. Ohm, and G. Brown- Guedira Haplotype structure and genetic diversity at Fusarium head bight QTLs in soft winter wheat germplasm. Proceedings of the 2007 National Fusarium Head Blight Forum. Kansas City, MO, December 2-4, 2007, page 214. M.S. Thesis: Bonin, Carolyn M.S. Thesis. Resistance to Fusarium head blight in wheat: Resistance QTL mapping and evaluation of resistance to kernel damage. 118 pages. (Form FPR07) 6

201 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Gretchen Kuldau Institution: Pennsylvania State University Address: Dept. of Plant Pathology 321 Buckhout Laboratory University Park, PA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 43,902 Factors and Mechanisms Favoring Deoxynivalenol Presence in Asymptomatic Wheat. USWBSI Individual Project(s) USWBSI Research Area * EEDF ARS Adjusted Award Project Title Amount Factors and Mechanisms Favoring Deoxynivalenol Presence in $43,902 Asymptomatic Wheat. Total Award Amount $ 43,902 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

202 PI: Kuldau, Gretchen USDA-ARS Agreement #: Project 1: Factors and Mechanisms Favoring Deoxynivalenol Presence in Asymptomatic Wheat. 1. What major problem or issue is being resolved and how are you resolving it? We are investigating the conditions that lead to asymptomatic grain containing >2 ppm deoxynivalenol (DON). Understanding this phenomenon is important for grain buyers and millers to protect them from inadvertently buying contaminated grain and for breeders to understand how this phenomenon interacts with disease resistance. We are studying the environmental conditions leading to asymptomatic, DON contaminated grain and we are conducting experiments to understand the in planta mechanisms. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A field study to evaluate the impact of moisture timing on disease severity, Fusarium Damaged Kernels (FDK), and DON was conducted using the susceptible variety Hopewell, and the moderately resistant varieties Truman and Valor. Our results demonstrate that moisture during the grain fill period can promote infection leading to disease, FDK and DON accumulation. We observed however that disease severity was lower than when moisture was available at anthesis. For the susceptible cultivar Hopewell, there was no significant difference between the ambient conditions and those with moisture during grain fill for disease severity or FDK but there was a significant difference for DON with the moisture at grain fill treatment having significantly more. Impact: These results confirm and expand on the research of others which point to the importance of moisture and infection during grain filling in addition to anthesis. A manuscript describing these results and those from a previous field season is complete and will be submitted for publication. This knowledge will help those preparing disease prediction models to refine their predictions and potentially incorporate information from weather events at more times during the season. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Data is now available using cultivars of wheat being developed now that indicates the importance of moisture during grain filling for disease development and DON accumulation. Additionally, our data provides the first evidence that moisture during grain filling can result in sound appearing grain that is contaminated with DON. Accomplishment: To understand the in planta mechanisms responsible for asymptomatic, contaminated wheat kernels we conducted several growth chamber experiments to assess the impact of temperature and to evaluate the movement of toxin and fungus in the wheat head. Wheat heads were point inoculated at a central floret and individual florets were harvested at various times after inoculation. For each spikelet, one floret was frozen for later chemical (Form FPR07) 2

203 PI: Kuldau, Gretchen USDA-ARS Agreement #: analysis and one was plated to check for fungal colonization. Three separate runs of this experiment were run using the spring wheat varieties Wheaton (susceptible) and Alsen (resistant). Wheat heads of both varieties were fully colonized by eight days after inoculation at 22 C. At 15 C Alsen heads were fully colonized at eight days while Wheaton heads were not fully colonized until 14 days. Colonization of Alsen occurred before Wheaton at 15 C despite being moderately resistant. This suggests that current methods used in resistance trials may be insufficient. Trials that assess multiple types of resistance might be more effective. Impact: This new knowledge may influence breeders and their decisions regarding resistance screening. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The chemical analysis of the florets is not complete so the full story is not yet known. Accomplishment: Single wheat florets from growth chamber experiments described above will be analyzed for DON and the fungus specific sterol ergosterol. The ergosterol measurement will give an indication of fungal biomass in the floret. A gas chromatography method was developed for this analysis. The method uses a standard extraction for DON followed by saponification of the wheat floret for extraction of ergosterol. The samples are derivitzed with HFBA and detected by electron capture. Mirex is used as an internal standard. DON elutes at 20.4 minutes and has a limit of detection of ng/ul. Ergosterol elutes at 47.7 minutes and has a limit of detection of 0.05 ng/ul. We have successfully detected and quantitated DON and ergosterol in single wheat florets. Preliminary data from a small number of wheat heads indicates that in susceptible varieties ergosterol and DON both accumulate at the point of inoculation and beyond. In the resistant variety however levels of DON were very high compared to ergosterol and ergosterol was restricted the point of inoculation while DON was seen above and below the point of inoculation. Impact: Ours is the first method to utilize gas chromatography with electron capture detection for quantitation and detection of DON and ergosterol in single wheat florets. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: A method now exists for the study of the movement of the fungus and the toxin in wheat heads. This will permit dissection of differences amongst varieties in terms of build up toxin and fungus and this data can be paired with information on disease symptoms. (Form FPR07) 3

204 PI: Kuldau, Gretchen USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Willyerd, K. T., De Wolf, E. D., Nita, M. and Kuldau, G. A Timing of Infection: The effects on Fusarium Head Blight severity and toxin accumulation in wheat kernels. Phytopathology 97:S123. (Oral presentation.) Willyerd, K. T., Boroczky, K., Kuldau, G. A A method for quantifying trichothecenes and ergosterol in single wheat florets using gas chromatography with electron capture detection. In: Canty, S. M., Clark, A., Ellis, D., and Van Sanford, D. Proceedings of the National Fusarium Head Blight Forum; 2007 Dec. 2-4; Kansas City, MO. Lexington, KY: University of Kentucky. pp 142. (poster) Willyerd, K. T., Nita, M., DeWolf, E. D., Kuldau, G. A Influence of infection timing on Fusarium head blight severity, wheat kernel damage and deoxynivalenol accumulation during a 2007 field study. In: Canty, S. M., Clark, A., Ellis, D., and Van Sanford, D. Proceedings of the National Fusarium Head Blight Forum; 2007 Dec. 2-4; Kansas City, MO. Lexington, KY: University of Kentucky. pp 143. (poster) (Form FPR07) 4

205 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Michael Lawton Institution: Rutgers, The State University of New Jersey Address: Biotech Center - Cook College 59 Dudley Road New Brunswick, NJ lawton@aesop.rutgers.edu Phone: ext. 223 Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 50,052 A Rapid Assay System for Trangenes that Confer Resistance to DON and FHB. USWBSI Individual Project(s) USWBSI Research Area * GET ARS Adjusted Award Project Title Amount A Rapid Assay System for Transgenes that Confer Resistance to $50,052 DON and FHB. Total Award Amount $ 50,052 7/13/08 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries

206 PI: Lawton, Michael USDA-ARS Agreement #: Project 1: A Rapid Assay System for Transgenes that Confer Resistance to DON and FHB. 1. What major problem or issue is being resolved and how are you resolving it? There is an urgent need in the field for germplasm and genes that can enhance resistance of wheat and barley to FHB and thereby prevent the accumulation of DON. Because transformation of wheat and barley is time- and resource-consuming, it is not possible to screen large numbers of transgenes genes for their activity against FHB in these systems. We have addressed this problem by developing a rapid and efficient gene assay system based on the recombinogenic plant Physcomitrella patens, which allows gene function to be rapidly and inexpensively assessed through the creation of gene knockout or overexpression lines. By exploiting this plant as a rapid assay system, we have been able to identify a number of genes that confer resistance to DON and to FHB. Genes that are effective in Physcomitrella have been passed on to our collaborators in the USWBSI program who can validate their performance in a transient assay for efficacy against FHB in wheat. Cumulatively, these linked assays constitute a research and development pipeline for gene discovery and deployment of novel anti-fhb genes in wheat. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have characterized a collection of genes for efficacy against DON and FHB in Physcomitrella. These genes define independent pathways for conferring resistance to FHB and DON and identify multiple targets for improving FHB-resistance in wheat through classical or molecular approaches. Wheat versions of these genes are also effective against FHB in Physcomitrella. Constructs designed to suppress the activity of these anti-fhb genes in wheat have been developed and are being assayed for efficacy against FHB in whole wheat plants. These studies allow highly performing anti-fhb genes to be selected for introduction into transgenic wheat plants. Impact: These developments constitute a research and development pipeline for screening genes for DON and FHB resistance. They provide a collection of genes that can be targeted for improvement by molecular or marker-assisted methods or through gene transfer. They also establish the R&D pipeline for the discovery and deployment of additional genes as these are uncovered from ongoing screens. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community has access to the anti-fhb genes that we have validated in Physcomitrella, along with an indication of their utility for improving crop plants. The scientific and agricultural communities can design better, more rational and robust strategies for improving FHB resistance based on the results of our mechanistic studies in Physcomitrella. These communities will also benefit from the establishment of a dedicated route-to-deployment for anti-fhb genes into wheat. (Form FPR07) 2

207 PI: Lawton, Michael USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Publications: Saidasan, H. and Lawton, M. (2007) In Proceedings of the 2007 National Fusarium Head Blight Forum (Eds, Canty, S. M., Clarke, A., Ellis, D. and Van Sanford, D.) University of Kentucky, Kansas City, MO, pp. 58. Lawton, M and Saidasan, H. (2008). Physcomitrella Pathogenesis. In The Moss, Physcomitrella, C, Knight, D. Cove and P.-F. Perroud, Eds. Annual Plant Reviews, Wiley-Blackwell, Oxford. In press Presentations: Genes that confer resistance to Fusarium. Second Annual Rutgers Mini-Symposium: Cultivating Traditions, Current Strengths and Future Frontiers, Feb January 7-8, 2008, Poster Presentation. Genes that confer resistance to Fusarium. National Fusarium Head Blight Forum, Kansas City, MO, Dec 2-4, Invited Speaker. Genes that confer resistance to Fusarium. National Fusarium Head Blight Forum, Kansas City, MO, Dec 2-4, Poster Presentation. Gene networks that control disease susceptibility in Physcomitrella patens, Dept. Chemistry, University of Sao Paulo, SP, Brazil, Dec 12, Invited Speaker. Mechanisms of moss deconstruction: Microbial Infection of Physcomitrella, Moss 2007, Korea University, Seoul, Korea, August Invited Presentation. Physcomitrella patens: a model genetic system for understanding susceptibility to Fusarium Head Blight. 5th Tripartite Symposium in Biotechnology and BioEnergy, New Brunswick, NJ. April 9-12, Poster presentation. (Form FPR07) 3

208 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: John Leslie Institution: Kansas State University Address: Department of Plant Pathology 4024 Throckmorton Hall Manhattan, KS Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Genetic Diversity and Genetic Mapping of Gibberella zeae. Title: FY07 ARS Award Amount: $ 32,629 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount PGG Genetic Diversity in and Genetic Mapping of Gibberella zeae. $32,629 Total Award Amount $ 32, July 2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

209 PI: Leslie, John USDA-ARS Agreement #: Project 1: Genetic Diversity in and Genetic Mapping of Gibberella zeae. 1. What major problem or issue is being resolved and how are you resolving it? A number of phylogenetic lineages (now 11) are known with G. zeae and have been formally described as phylogenetic species. Considerable genetic diversity in gene sequence exists between the lineages, but the lineages are not reproductively isolated under laboratory conditions suggesting that the lineages may represent (formerly) isolated populations and not distinct species. Published sequences for lineage-associated genes are from a relatively few strains. We sequenced four genes in G. zeae strains from the United States to determine whether the published sequences were representative of those present in the field. These same genes also have been sequenced from 472 isolates representing multiple lineages from South America, which should enable a comparison of these populations as well. From these sequences we will be able to determine the relatedness of these populations and determine if there are unique sequences present in the US populations of this fungus. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Previously, Amplified Fragment Length Polymorphisms (AFLPs) were used to evaluate the genetic diversity of 523 isolates from North America and assign them to one of 503 haplotypes (presumptive clones) belonging to F. graminearum lineage 7 (F. graminearum sensu stricto). We sequenced four genes [Mating Type (MAT), TRI-101, Reductase (RED), and β- tubulin (TUB2)] from strains representing ~350 of these haplotypes. In all cases, multiple alleles were found (some not previously reported): RED 19 (15) alleles, TUB2 3 (1) alleles, MAT 7 (4) alleles, and TRI (7) alleles. All alleles present at a frequency > 1% were recovered from multiple locations, suggesting that these populations are wellmixed. The new alleles represent 35% of the total RED sequences, < 1% of the TUB2 sequences, 2.2% of the MAT sequences and 9.4% of the TRI101 sequences. Genetic relatedness networks for these four genes (using the available lineage tester strains to represent nine of the eleven described lineages) are not homologous with regards to the association of the lineages. This lack of concordance between gene genealogies is consistent with a hypothesis that these lineages are distinct but that all form a part of a single species and should not yet be accorded species status. The polytomies that characterize these genealogies also are consistent with all nine lineages being members of the same species rather than members of a number of different species. The data from the North American strains blurs the differences between lineage 7 and the other lineages by identifying additional alleles that are intermediate between the described alleles and those in lineage 7. These data also provide evidence for intragenic recombination at some loci, which when combined with the relatively large number of alleles suggests that these populations are relatively old and probably stably established. Impact: The taxonomic status of G. zeae/f. graminearum is of critical importance for plant quarantine and trade measures. If there are a number of species then each must be treated separate- (Form FPR07) 2

210 PI: Leslie, John USDA-ARS Agreement #: ly and the presence/absence of a particular species can be used as a non-tariff trade barrier. Our results strongly suggest that while isolated populations of F. graminearum exist, these populations are not reproductively isolated and should be recognized as portions of a single, large, diverse species rather than as nine discrete entities. Sequencing of additional DNA variants is consistent with this conclusion as alleles that are missing from the initial alignments are found and begin to blur the distinctness of the various groups. Recognition of a single species does not materially impact the plant quarantine regulations currently in place, nor alter the application/implementation of current trade practices. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? The controversy over species or lineage as the proper designation for the groups observed within F. graminearum has been ongoing. These data suggest that a single species is most appropriate since the differences between lineages are now less distinct than they were in the past. They also suggest that lineage/speciation techniques that rely on one or a few single nucleotide polymorphisms for their diagnostic capability have an error rate that has yet to be quantified, and that these techniques should not be used beyond the research level until the error rates are determined. (Form FPR07) 3

211 PI: Leslie, John USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Books: 1. Leslie, J. F., R. Bandyopadhyay & A. Visconti, eds Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade. CABI, Kew, UK. 476 pp. Refereed journal articles: 1. Hornok, L., C. Waalwijk & J. F. Leslie Genetic factors affecting sexual reproduction in toxigenic Fusarium species. International Journal of Food Microbiology 119: Leslie, J. F., L. L. Anderson, R. L. Bowden & Y.-W. Lee Inter- and intra-specific genetic variation in Fusarium. International Journal of Food Microbiology 119: Ramirez, M. L., M. M. Reynoso, M. C. Farnochi, J. F. Leslie & S. N. Chulze Population genetic structure of Gibberella zeae from wheat in Argentina. Food Additives and Contaminants 24: Bentley, A. R., J. F. Leslie, E. C. Y. Liew, L. W. Burgess & B. A. Summerell Genetic structure of Fusarium pseudograminearum populations from the Australian grain belt. Phytopathology 98: Lee, J., J. F. Leslie & R. L. Bowden Expression and function of sex phermones and receptors in the homothallic ascomycete Gibberella zeae. Eukaryotic Cell 7: Lee, J., J. E. Jurgenson, J. F. Leslie & R. L. Bowden Alignment of genetic and physical maps of Gibberella zeae. Applied and Environmental Microbiology 74: Bentley, A. R., M. G. Milgroom, J. F. Leslie, B. A. Summerell & L. W. Burgess Spatial aggregation in Fusarium pseudograminearum populations from the Australian grain belt. Plant Pathology (in press). 8. Bowden, R. L., I. Fuentes-Bueno, J. F. Leslie, J. Lee & Y.-W. Lee Methods for detecting chromosomal rearrangements in Gibberella zeae. Cereal Research Communications (in press). 9. Leslie, J. F., and R. L. Bowden Fusarium graminearum: When species concepts collide. Cereal Research Communications (in press). Book chapters: 1. Bandyopadhyay, R., R. A. Frederiksen & J. F. Leslie Priorities for mycotoxin research in Africa identified by using the nominal group technique. In: Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade (J. F. Leslie, R. Bandyopadhyay & A. Visconti, eds.), pp CABI, Kew, UK. 476 pp. 2. Shelton, B. G. & J. F. Leslie Comparative risks of airborne and foodborne molds and mycotoxins. In: Mycotoxins: Detection Methods, Management, Public Health and Agricultural Trade (J. F. Leslie, R. Bandyopadhyay & A. Visconti, eds.), pp CABI, Kew, UK. 476 pp. (Form FPR07) 4

212 PI: Leslie, John USDA-ARS Agreement #: Abstracts and meeting presentations: 1. Anderson, L. L., Y.-W. Lee, R. L. Bowden & J. F. Leslie Relationships between alleles at lineage diagnostic loci in Fusarium graminearum. Fungal Genetics Newsletter 54(Suppl.): Lee, J., R. L. Bowden & J. F. Leslie Pheromone functions in Gibberella zeae. Fungal Genetics Newsletter 54(Suppl.): Lee, J., J. F. Leslie & R. L. Bowden Functions of the sex pheromones of Gibberella zeae. Proceedings of the 2007 National Fusarium Head Blight Forum (Kansas City, Missouri): Reynoso, M. M., M. L. Ramirez, J. F. Leslie & S. N. Chulze Trichothecene chemotypes of isolates of Gibberella zeae recovered from wheat in Argentina. Proceedings of the 2007 National Fusarium Head Blight Forum (Kansas City, Missouri): 34. Dates and locations of invited presentations by Dr. Leslie that contained information from this project but for which there is no published abstract: 1. Bioforsk, Ås, Norway 03/ College of Life Sciences, Dalian Nationalities University, Dalian, China 04/ Shenyang Agricultural University, Shenyang, China 04/ Faculty of Agricultural & Life Sciences, Seoul National University, Seoul, Korea 05/ FABI, University of Pretoria, Pretoria, South Africa 11/ Norwegian National Veterinary Institute, Oslo, Norway 04/ Faculty of Agricultural & Life Sciences, Seoul National University, Seoul, Korea 05/08. (Form FPR07) 5

213 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Janet Lewis Institution: Michigan State University Address: Department of Crop & Soil Sciences 382 PSSB East Lansing, MI Phone: ext. 185 Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 82,257 Development of FHB Resistant Soft White Wheat Varieties for Michigan and similar Environments. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Development of FHB Resistant Soft White Wheat Varieties for $82,257 Michigan and similar Environments. Total Award Amount $ 82,257 July 15, 2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

214 PI: Lewis, Janet USDA-ARS Agreement #: Project 1: Development of FHB Resistant Soft White Wheat Varieties for Michigan and similar Environments. 1. What major problem or issue is being resolved and how are you resolving it? Soft white winter wheat (SWWW) is a significant element of Michigan agriculture- both on the farm and in the food processing industry. Kellogg s and other major cereal companies use processed soft white wheat with high-bran content in many of their products. The high concentration of wheat in such cereal foods results in lower maximum tolerances for deoxynivalenol (DON) in soft white wheat. In addition, the higher concentration of DON in bran, vs. the flour, makes it even more critical to have low DON levels in soft white wheat. SWWW cultivars with effective resistance to FHB are insufficient. In addition, SRWW is now being grown by a large proportion of MI farmers. MI has also begun working to ensure improved levels of FHB resistance in SRWW varieties that perform well in the state of MI. We have been working to resolve this problem through regular FHB field screening, exchange of germplasm with other breeders, and crossing to incorporate FHB resistance, including resistance with known QTL. In addition, samples are being collected and sent to MN for deoxynivalenol (DON) testing. Our achievements are highlighted below. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment (1): Release of MSU Line E2017. E2017 is a high yielding soft white wheat variety with significantly improved resistance to FHB (visual symptoms) and DON contamination, in comparison with the highest yielding soft white winter wheat varieties recently evaluated in MI. In addition, in comparison with Caledonia (a lower yielding variety in MI) the most widely grown soft white wheat variety in MI, E2017 shows improved FHB resistance, and similar DON levels. MSU Line E1009, another soft white wheat line with lower DON levels (though not lower visual symptoms) in comparison with other high yielding MSU releases, was licensed for commercialization. Impact (1): The availability of these lines gives farmers in MI the choice of a soft white wheat variety with lower FHB and/or DON contamination. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (1): The availability of these lines gives farmers in MI the choice of a high yielding soft white wheat variety with lower FHB and/or DON contamination. The production of these lines, especially E2017, will lessen the risk of FHB that the soft white wheat industry is facing. (Form FPR07) 3

215 PI: Lewis, Janet USDA-ARS Agreement #: Accomplishment (2): Crossing to Develop High Yielding FHB Resistant Varieties: In the spring of 2008, 151 crosses two and three-way crosses were made to develop high yielding lines with improved FHB resistance. In each cross, at least one of two/three parents had moderate to high levels of resistance to FHB visual symptoms and/or reduced DON levels. Approximately 50% (seventy-three) of these crosses had at least one of three parents with FHB resistance associated with known QTL (see table below for parents with known QTL for resistance). Other parents used as FHB resistance donors include Truman, MO , and VA03W-409, MSU Line E2017, MSU Line E2043, and MSU Line E6039. FHB resistant parent in simple crosses, 2007 Grain color Pedigree Information (Known source of FHB resistance in bold) QTL identified (small grains genotyping center) DON levels (ppm), 2006 MSU harvest. Years in NUWWSN MSU Line E6001 Red Pioneer 25W60/CJ9306 3BS, hetero for 2DL /07, 07/08 MSU Line E6002 White VA96W-403-WS/CJ9403 2DL /07, 07/08 MSU Line E6003 White VA96W-403-WS/W14 3BS and 2DL /07, 07/08 FHB12 (developed by Mark Sorrells) White Caledonia with 3BS gene added by MAS 3BS (according to Mark Sorrells) Results of testing of E6001, E6002 and E6003 in the 2006/07 NUWWSN confirmed, on a regional level, that these lines have good resistance to FHB (lines E6002 and E6003 ranked first and third, respectively, for FHB Index among 60 entries from multiple collaborators - Truman was ranked 2 nd ). Impact (2): The use of FHB resistant lines with known QTL for resistance will allow us to use Marker Assisted Selection (MAS) as early as possible (F1 for 3-way crosses, F2 for 2-way crosses) to identify the most likely FHB resistant lines. In addition, it is known that mapping efforts are underway for Truman, and therefore MAS will likely be possible for Truman crosses in the near future. Other lines, such as MSU E2017, E2043, E6039, VA03W-409 and MO are more likely to produce a large number of lines with good agronomic qualities, since these have been selected for performance in addition to FHB resistance. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (2): As a result of these crosses, our development and selection of FHB resistant lines is more rapid. Therefore, we are more likely to produce with FHB resistance from these crosses. Accomplishment (3): Field Evaluations for FHB resistance (2008). Many different nurseries are being evaluated for FHB resistance in an artificially inoculated (grain spawn and misted) field at Michigan State University. In addition to elite MSU germplasm, we are evaluating the state variety trial, regional nurseries, and MSU early generation germplasm. (Form FPR07) 3

216 PI: Lewis, Janet USDA-ARS Agreement #: Trial Name # Entries # Replication Evaluated State Performance Trial 76 3 Advanced Yield Trial 20 3 Preliminary Yield Trials (combined) 77 2 Northern Uniform Winter Wheat Scab Nursery 64 3 Northern Preliminary Winter Wheat Scab Nursery 64 3 Uniform Eastern Soft Red Winter Wheat Nursery 45 2 Uniform Eastern Soft White Winter Wheat Nursery 25 2 F4 White MSU Breeding Germplasm F4 Red MSU Breeding Germplasm 64 1 F3 Red MSU Breeding Germplasm Our FHB symptoms this year were very good. Disease levels between the genotypes varied from less than 10% to greater than 95% at 21 days post anthesis. Preliminary examination of the MSU germplasm suggests that 37 MSU genotypes (in addition to E2017 and E1009 discussed above) between the Preliminary, Advanced, and State Performance Trials showed moderate levels of resistance, 25 of which are soft white lines. Regarding the early generation materials, single plot data suggests that 23 (6.5%) showed very good levels of resistance, while 69 (19.5%) showed moderate levels of resistance. For the F4 and F3 red varieties combined, 38 (16.5%) showed very good levels of resistance, while an additional 75 (33%) showed moderate levels of resistance. All these genotypes (early generation and elite) are also planted in MSU s breeding nursery, where they are evaluated for agronomic performance. In addition to the data shown above, 22 lines with good FHB performance were identified previously, and are now in the breeding nursery field for selection based on agronomic performance. Impact (2): We have identified 264 MSU genotypes (both white and red), between the elite yield trials and early generation germplasm that have good potential for the development of varieties with improved FHB resistance. These genotypes must still be selected for agronomic performance. In addition, we have evaluated genotypes from multiple breeding programs, both private and public. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (2): Valuable data on FHB performance in MI for regional and state trials. In addition, genotypes evaluated in MI can be selected by the MI breeder as parents for crossing. Germplasm development shows that varieties with improved FHB resistance are in the pipeline at MSU and previous crossing efforts have effectively incorporated FHB resistance into the breeding populations at MSU. (Form FPR07) 4

217 PI: Lewis, Janet USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Presentations were made in various venues (mostly industry and extension/farmer meetings) by the PI during FY07 regarding the MSU breeding program, with emphasis on development of varieties with improved FHB resistance. These include: Scientific Meetings: J. Lewis, L. Siler, S. Hammar, (2008). MSU s Wheat Breeding Program; Current Status and Future Outlook. Soft Wheat Quality Laboratory 55 th Research Review Conference. March Wooster, OH. Industry Meetings, invited presentations (overviews/updates of wheat program to date): J. Lewis, L. Siler, S. Hammar (2008) MSU Wheat Breeding Program Update. Michigan State Miller s Association 143 rd Summer Meeting. June Mackinac Island, MI. J. Lewis, L. Siler, S. Hammar (2008) MSU Wheat Breeding Program Update. Soft White Wheat Endowment Meeting, June 11. Michigan State University, East Lansing, MI. J. Lewis, L. Siler, S. Hammar (2008) MSU Wheat Breeding and Genetics Program. Michigan Crop Improvement Association Annual Meeting. March 6. Bath, MI. J. Lewis, L. Siler, S. Hammar (2008) MSU Wheat Breeding and Genetics Program. Michigan State Miller s Association, 143 rd Winter Meeting, January 29, Michigan State University, East Lansing, MI L. Siler, G-L. Jiang, J. Lewis (2007) MSU Wheat Breeding Program. Michigan Farm Bureau Feed Grain, Oilseeds, and Wheat Advisory Committee Meeting. August 1. Eaton County, MI. L.Siler, G-L. Jiang, J. Lewis (2007) MSU Wheat Breeding Program Report. Michigan State Miller s Association, 142 nd Summer Meeting. June 16. Traverse City, MI. Extension Meetings, invited presentations (overviews/updates of wheat program to date): J. Lewis, L. Siler. MSU Wheat Breeding Program Yield Trials. Lenawee/Monroe County, MI, Extension Meeting. June 23. Hosted by Ned Birkey, MSU Extension Agent. J. Lewis, L. Siler, S. Hammer (2008) FHB Sceening Nursery Field Presentation. Ingham County, MI Extension Meeting. June 25. Hosted by Dan Hudson, MSU Extension Agent. (Form FPR07) 5

218 PI: Lewis, Janet USDA-ARS Agreement #: J. Lewis, L. Siler, S. Hammar (2008). Visit of Dr. Jonas Mugabe Debute Director Istitut des sciences agronomiqes du Rwanda to the MSU FHB screening nursery. June 20. East Lansing, MI. J. Lewis. L. Siler. S. Hammar. (2008) Wheat; A Look Into the Crystal Ball, MSU Wheat Breeding and Genetics Program. Progressive Farmer Winter Program Series. February 27. Clinton County, MI. Hosted by Marilyn Thelen, MSU Extension Agent. J. Lewis, L. Siler, S. Hammar. (2008) MSU Wheat Breeding and Genetics Program. Lenawee/Monroe County, MI, Extension Meeting. February 12. Hosted by Ned Birkey, MSU Extension Agent. J. Lewis, L. Siler (2007) Wheat Breeding for Michigan Agronomy Update Meeting. December 19, Michigan State University, East Lansing, MI (Form FPR07) 6

219 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Anne McKendry Institution: University of Missouri Address: Department of Agronomy 106 Curtis Hall Columbia, MO Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Fusarium Head Blight Research in Winter Wheat. Title: FY07 ARS Award Amount: $ 116,352 USWBSI Individual Project(s) USWBSI Research Area * HGG VDUN ARS Adjusted Award Project Title Amount Mapping QTL Associated with Scab Resistance in the Soft Red Winter Wheat Truman. $39,024 Accelerating the Development of Scab Resistant Soft Red Winter Wheat. $ 77,328 Total Award Amount $ 116,352 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

220 PI: McKendry, Anne USDA-ARS Agreement #: Project 1: Mapping QTL Associated with Scab Resistance in the Soft Red Winter Wheat Truman. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) resistance in the soft red winter wheat variety Truman developed and released at the University of Missouri is broad-based, having excellent levels of types I and II resistance as well as good kernel quality retention and low DON under heavy disease pressure. Little, however, is understood about the genetics of this source of resistance, other than that haplotype data suggests it differs from the resistance in Sumai 3 and its derivatives as well as the resistance in Ernie. In that Truman has good levels of all four types of resistance, this project will involve several steps. In 2007/2008, our objectives were: (1) identify QTL associated with type II resistance to FHB in a set of 250 F 2:8 recombinant inbred lines developed from the cross Truman/MO ; (2) Phenotype type I resistance to FHB in Truman using the above mapping population; (3) phenotype kernel quality retention in Truman following field inoculations of the above mapping population; (4) phenotype the above mapping population for DON levels following greenhouse and field inoculation with FHB. 1. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A replicated (3 replication; randomized complete block design) greenhouse phenotyping experiment was conducted on a set of 250 recombinant inbred lines (RILs) developed from the cross Truman/MO in 2007/08. For each RIL, eight plants per RIL per replication were phenotyped (total 6000 plants). The population was also increased for field screening in the 2008/09 crop season. Analysis of the data indicated that resistance is quantitative and distributions which were skewed towards resistance, suggest both additive and dominant components of the phenotypic variance. Diallel analysis indicated that Truman had among the best breeding values for FHB resistance of lines evaluated. Type II resistance was highly heritable (72%) and appears to be conditioned by a minimum of 3 genes. Replicated (2 replications; RCBD) field evaluations have been completed for type I resistance, percentage of Fusarium damaged kernels (FDK) and DON content of the harvested grain. Whole genome polymorphisms between Truman and MO will be initiated this fall in preparation for mapping resistance in this background. Impact: Truman, the resistant check in the Northern Scab Nursery, is arguably the most FHB resistant cultivar available to growers in the soft red winter wheat region. It is widely adapted and has excellent yield potential along with excellent test weight and stripe rust resistance. It has immediately lessened economic losses due to FHB in regions where it is grown. Haplotying indicates that is potentially carries FHB alleles that differ from others now in use. Once markers are identified and published, this will be a valuable resource for marker-assisted selection. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Once identified and published through the peer-reviewed literature, markers from Truman will be an extremely valuable addition to the set currently in use for marker-assisted-selection. (Form FPR07) 2

221 PI: McKendry, Anne USDA-ARS Agreement #: Project 2: Accelerating the Development of Scab Resistant Soft Red Winter Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight, in wheat continues to be an important problem in the north-central region of the United States. This ongoing project has focused largely on the exploiting native resistance identified in Missouri wheat germplasm; an approach we believe accelerates the development and release of FHB resistant cultivars. In 2007our objectives were: (1) the continued identification and verification of useful sources of FHB resistance in the Missouri wheat breeding program; (2) the verification of potentially useful sources of FHB resistance through evaluation of both the Preliminary and Advanced Northern Uniform Winter Wheat Scab Nurseries, and the Southern Uniform Winter Wheat Scab Nurseries; (3) the incorporation of new sources of resistance, as they are verified, into elite Missouri soft red winter wheat breeding lines; (4) preliminary genetic characterization through haplotyping of Missouri resistant breeding germplasm; (5) evaluation of lines with diverse sources of resistance derived in 2006 from mass selection populations; (6) to convey information on scab resistance of commercially available wheat cultivars through evaluation of the 2008 Missouri Winter Wheat Performance Tests. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: A severe FHB outbreak provided heavy disease pressure for Missouri trials and a good test of newly developed materials. Highly susceptible lines had more than 80% loss. Of 250 advanced lines carrying only native resistance, 112 (45%) had resistance ratings assessed as the Fusarium head blight index (incidence x severity) that were equal to or better than Truman (<10%). Of 1700 preliminary lines with complex pedigrees involving both native and exotic FHB sources of resistance, 534 (31.5%) had resistance levels similar to or better than Truman. Among the most resistant lines are lines that appear to combine the resistance in Truman (or its sibs) with that in Ning 7840, lines that combine the resistance in Ernie with that in Sumai 3, and lines that could potentially contain resistance from Truman, Sumai 3 and Ernie in one background. Resistant lines will be culled based on pedigree, agronomic performance (yield, test weight, resistance to other pathogens) and grain quality (milling and baking). They will then be advanced into replicated state-wide testing. Each line that is retained will be moved into the greenhouse/field shuttle screening program to confirm resistance and concurrently screened with known SSR markers to determine FHB gene combinations. Impact: The release of FHB-resistant cultivars is the primary goal of the USWBSI. Our program has had considerable success focusing on native resistance through screening and recombination in adapted backgrounds. The verification of resistance in these lines under good levels of natural infection over a broad heading window will accelerate our decision making on both retention of lines, and elimination of lines, in the breeding program. Because of the adapted backgrounds in these lines, they have most of the necessary attributes for release and will immediately lessen the devastating impact of FHB in Missouri and surrounding states. (Form FPR07) 3

222 PI: McKendry, Anne USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These lines, which have been developed within the Missouri program will provide the agricultural community we serve in Missouri and the surrounding states with continued access to FHBresistant wheat varieties. When these lines are shared, through the uniform scab nursery system, breeders in other states will also be able to utilize these resources, either as parents or directly as cultivars through joint release. Accomplishment 2: Dissemination of information on FHB resistance to growers is a newly identified goal of the USWBSI. One mechanism by which that can be done is to evaluate all commercial varieties available to growers and extend information on their respective resistance to FHB through field day events and through publication (both web-based and hard copy) of the data in wheat variety test publications. In 2007/2008, we evaluated type II resistance for 64 entries in the Missouri Winter Wheat Performances tests under greenhouse conditions, and the FHB field scab index (incidence x severity) in our FHB inoculated nursery. In addition, a significant outbreak in Missouri enabled us to take FHB resistance notes under natural infection at two additional Missouri locations. Less than 10% of entries tested had good FHB resistance. As anticipated, the best public lines in these commercial trials were Truman, Bess and Roane. These data will be published in the 2008 Missouri Winter Wheat Performance Tests Special Report and on the Agricultural Education Bulletin Board under crop performance testing for soft red winter wheat. Impact: These data will provide growers with previously unavailable information on the FHB resistance of commercial cultivars. The availability of these data, particularly in a year in which FHB has been a problem for growers, and the availability of resistant varieties in the test including Truman, Bess, and Roane, should move growers decision making on variety selection towards selection of varieties with FHB resistance. This will have an immediate impact lessening the devastating effects of FHB in Missouri and surrounding states where the data will be available. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Fusarium head blight resistance data on commercial cultivars developed by private companies, and on brands that are widely used has not been widely available to Missouri growers. As a consequence of the publication of these FHB data, growers will be able to make more informed decisions on variety selection that should, with adoption of resistant varieties, less their risk of crop loss associated with FHB. (Form FPR07) 4

223 PI: McKendry, Anne USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Abate, Z.A., Liu, S., McKendry, A Quantitative trait loci associated with deoxynivalenol content and kernel quality in the soft red winter wheat Ernie. Crop Sci. 48: Abate, Z.A., D.N. Tague, and A.L. McKendry Diallel analysis of Fusarium head blight resistance in genetically diverse winter wheat germplasm. American Society of Agronomy Annual Meeting Abstracts. New Orleans, LA. Nov. 4-8, Dowell, F.E., E.B. Maghirang, P.S. Baenziger, A.L. McKendry, E.A. Milus, P. Horevaj, S.N. Wegulo, J.A. Breathnach Applying single kernel sorting technology to developing scab resistant lines. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford D. (Eds) Proceedings of the National Fusarium Head Blight Forum. 2-4 December 2007; Westin Crown Center, Kansas City, MO. Liu, S., C.A. Griffey, A.L. McKendry, and M.D. Hall Meta-analyses of QTL associated with Fusarium head blight resistance. In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford D. (Eds)Proceedings of the National Fusarium Head Blight Forum. 2-4 December 2007; Westin Crown Center, Kansas City, MO. McKendry, A.L., Breeding For Fusarium Head Blight Resistance in Winter Wheat - What's Ahead. Presented at the 2007 US Wheat and Barley Scab Forum, in Kansas City Missouri. (Associated Abstract In: Canty, S.M., Clark, A., Ellis, D., and Van Sanford D. (Eds) Proceedings of the National Fusarium Head Blight Forum. 2-4 December 2007; Westin Crown Center, Kansas City, MO) Under review: Abate, Z.A., A.L. McKendry Diallel analysis of Fusarium head blight resistance in genetically diverse winter wheat. Under review: Euphytica. (Form FPR07) 5

224 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Marcia McMullen Institution: North Dakota State University Address: Department of Plant Pathology Walster Hall, Rm 303 Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 86,201 Collaborative Studies of Fungicides and Application Techniques for Improved Control of Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * CBCC CBCC EEDF ARS Adjusted Award Project Title Amount Uniform Evaluation of Fungicides for Control of FHB in Multiple ND Grain Classes. $13,506 Uniform Evaluation of IPM Practices for FHB in Multiple Grain Classes, ND. $ 43,427 Greenhouse Studies of DON Development in Spring Grains as Influenced by Infection Timing, Moisture, and Temperature. $ 29,268 Total Award Amount $ 86,201 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

225 PI: McMullen, Marcia USDA-ARS Agreement #: Project 1: Uniform Evaluation of Fungicides for Control of FHB in Multiple ND Grain Classes. 1. What major problem or issue is being resolved and how are you resolving it? Fungicides continue to be one of the important management strategies for wheat and barley producers to reduce yield and quality losses due to FHB infection. Uniform tests of fungicides across multiple grain classes and environments provides information on how registered or experimental fungicides perform on these grain classes common in North Dakota, and multiple testing sites help assure that at least one or more sites may have disease development. In 2007, uniform fungicide tests (using a common protocol developed by the USWBSI management group) were done on spring wheat, durum wheat, and barley, across three sites in ND, sites representing different climatic and soil environments. Data from these trials has been instrumental in providing up-to-date management information to producers and in getting new fungicides registered. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Relatively dry weather conditions in ND in 2007 did not favor FHB infection in most commercial fields, but research trials in this study provided added mist and/or inoculum that allowed good separation of efficacy of products on spring wheat, durum, and barley. For example, results indicated that Proline, Prosaro, and a high rate of Caramba fungicide gave superior control of FHB and reduction in DON than other products tested. Compilation of results over multiple locations showed a 10 to 15% greater reduction in DON levels and up to a 10 percent greater yield increase with these better products versus the standard used, Folicur, while Folicur provided an average 15% greater reduction in DON and 6% greater yield than the other product tested, Topguard. Impact: Data from these ND fungicide trials, as well as from uniform trials across the US in other grain classes, contributed to the data base that resulted in EPA registrations of the best products available for FHB control. Proline was registered in 2007 and Folicur and Caramba were registered in Prosaro is a combination product of Proline and Folicur, so successful registration of both of these products allowed Prosaro registration in 2008, as well. The full registration of Folicur, which is off-patent, also opened the door for registration of generic products, six of which currently are now labeled in ND. Increased product availability means increased competition, which has given producers more options at reduced product price. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: As indicated in the impact statement, they now have three new fungicides available that are superior for controlling FHB and reducing DON. Under moderate FHB pressure, these products may provide a reduction of DON that meets the grain industry standard, plus provide up to 10-15% yield increase over other products tested. (Form FPR07) 2

226 PI: McMullen, Marcia USDA-ARS Agreement #: Project 2: Uniform Evaluation of IPM Practices for FHB in Multiple Grain Classes, ND. 1. What major problem or issue is being resolved and how are you resolving it? A number of management strategies have been demonstrated to reduce FHB and DON, strategies such as use of fungicides, crop rotation, tillage, and tolerant cultivars. However, under severe epidemics, an individual strategy used alone, or even use of two strategies, will not reduce disease severity and DON to levels required by the grain industry. A combination of strategies, each building upon the other, is required for achieving the best management of FHB and for obtaining quality grain. The purposes of this collaborative effort, within ND and across other states and grain classes are: 1. Demonstrate that integrated management is the most effective means of reducing losses to FHB/DON 2. Increase grower adoption of integrated strategies by demonstration of their effectiveness in a wide range of environments 3. Decrease the risk of the development of FHB epidemics and thus reduce losses to FHB and DON. In ND in 2007, we had a multi-site, multi-grain class study to evaluate the benefits of combining previous crop, host resistance and fungicides for FHB/DON management. The sites and grain classes were: hard red spring wheat at Fargo with 3 strategies; winter wheat at Lisbon and Prosper with variety x fungicide strategies; spring wheat at Carrington with variety x fungicide strategies, durum wheat at Minot with variety x fungicide strategy, durum at Langdon with all three strategies, and barley at Fargo and Langdon with all three strategies. All trials were established under natural rainfall and inoculum conditions (i.e. no mist-irrigation and no inoculation). Under these locations and weather conditions of 2007, successful evaluation of strategies for FHB control and DON reduction were achieved with the spring wheat studies in Fargo and Carrington, the winter wheat studies in two locations, the durum location in Langdon, and the barley study in Langdon. Too dry of environmental conditions at Minot, and field drown out of barley in Fargo, prevented successful FHB evaluations at these locations, indicating the value of multiple locations to obtain data for the research protocol. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Successful evaluation of use of multiple strategies was achieved on four spring grain classes in ND. Results showed that multiple strategies improved FHB control, DON reduction, and yield and quality parameters over single strategies, even with some of the more FHB resistant spring wheat varieties available. These results were presented at two invited presentations (Canadian and US Fusarium workshops), as well as presented at many producer meetings in the region. Winter wheat results were published in the Agronomy (Form FPR07) 3

227 PI: McMullen, Marcia USDA-ARS Agreement #: Journal. Our data helps meet the goals 1 and 3 of this coordinated project and we need to further measure grower adoption of the strategies. Impact: Growers have been provided information demonstrating that multiple strategies are most effective. For example, an FHB susceptible durum variety planted into hard red spring wheat stubble, and not treated with a fungicide, had 3x higher FHB field severity, 3x higher DON levels, a reduction in yield of 19 bushels, and a 2.8 lb lighter test weight than when a more FHB resistant durum cultivar had been planted on canola ground and treated with an effective fungicide at flowering. No single strategy in this study approached the gains made by multiple strategies. In spring wheat at Carrington, an FHB resistant cultivar and fungicide treatment resulted in a 10x reduction in FHB field severity and a 13 bushel yield increase over using a susceptible cultivar and no fungicide, while use of a resistant cultivar alone resulted in an 8x reduction in FHB field severity and an 8 bushel yield increase. Similar results were observed in the other grain classes where FHB developed. Surveys by the ND Wheat Commission indicating varieties grown indicate that growers are quickly adopting use of more FHB resistant varieties, and recent availability of superior fungicides may indicate more use when FHB disease forecasting sites indicate that fungicides are warranted. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Data that supports the use of multiple strategies improve FHB control. Integrated strategies have been demonstrated in Additional tests in 2008 will show how these multiple strategies work with possibly different environments and/or different varieties and fungicide products available. Results of individual trials in 2007 may be found at: North Dakota State University Small Grain Fungicide Field Trials compilation. NDSU Dept. of Plant Pathology, Fargo, ND. 82 pages. On-line at: (Form FPR07) 4

228 PI: McMullen, Marcia USDA-ARS Agreement #: Project 3: Greenhouse Studies of DON Development in Spring Grains as Influenced by Infection Timing, Moisture, and Temperature. 1. What major problem or issue is being resolved and how are you resolving it? Deoxynivalenol (DON) accumulation in wheat has been shown to be positively correlated with visible scab, but correlations between disease severity and DON levels are not always high. DON accumulation has been reported in asymptomatic winter wheat, raising questions about how time of infection, environmental conditions, or genetics influence DON production. We conducted greenhouse studies in 2003 and 2004 with hard red spring wheat that indicated that single postanthesis (Feekes 10.54) inoculations with Fusarium graminearum did not result in appreciable visible symptoms of FHB, but we did not measure DON levels. New cultivars of spring wheat are now commercially grown in ND that have Sumai-3 germplasm as a source of resistance, and the effect of timing of infection on these also need to be evaluated. Data from this research has several implications. For example, if late infections and prolonged moisture increase risk of DON in certain grain classes or cultivars, this information may be used: 1) to incorporate weather prediction into current FHB forecasting models for spring and winter wheat; 2) to prompt further evaluation of post-flowering fungicide efficacy and provide data on how close to fungicide pre-harvest intervals may fungicides be applied to be effective and yet follow label restrictions on these intervals (current superior FHB fungicides have a 30 day PHI); and 3) to assist breeders in incorporating DON resistance into their germplasm. Greenhouse experiments on two varieties of each of two wheat classes, hard red spring wheat and durum wheat, at four infection timings (Feekes 10.5, 10.51, and 11.1) and three misting durations (intermittent mist for 2 days, 5 days or 10 days following inoculation) were done in a controlled greenhouse environment at NDSU during the winter of At least two trials with each variety per grain class (representing two levels of FHB resistance) were done over the course of the winter/spring. FHB was assessed at soft dough stage and then plants were allowed to grow to maturity for hand harvesting and DON determination. DON, 3ADON, 15ADON and Nivalenol were measured by the NDSU Toxicology Laboratory, using gas chromatography and electron capture techniques. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Trials evaluating the effect of inoculation timing and mist duration were successfully completed on Glenn spring wheat (moderately resistant) and Trooper spring wheat (susceptible), and on Monroe durum (susceptible) and Divide durum (best available resistance among durums). Although all data has not been analyzed, preliminary results indicate that late inoculations produce substantial DON in more susceptible cultivars if mist duration is extended. Long durations of wetness are critical for DON development at all growth stages in the more resistant varieties. (Form FPR07) 5

229 PI: McMullen, Marcia USDA-ARS Agreement #: Impact: Data gathered from these trials will indicate the relative risk of DON accumulation in two spring grain classes when extended periods of moisture occur, no matter what the growth stage of infection, and provide data that will allow development of management strategies for possible late infections. Data accumulated also will indicate the presence or absence of DON derivatives. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Data indicating that late infections may be as damaging as infections at flowering, if extended periods of moisture occur following flowering. Management strategies still have to be developed for these late infections, or breeders will need to consider these late infections in their breeding programs. (Form FPR07) 6

230 PI: McMullen, Marcia USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Refereed: Ransom, J.K., and McMullen, M.V Yield and disease control on hard winter wheat cultivars with foliar fungicides. Agronomy J. 100: Paul, P.A., Lipps, P.E., Hershman, D.E., McMullen, M.P., Draper, M.A., and Madden, L.V A quantitative review of tebuconazole effect on Fusarium head blight and deoxynivalenol content in wheat. Phytopathology 97: R. R. Burlakoti, S. Ali, G. A. Secor, S. M. Neate, M. P. McMullen, and T. B. Adhikari Genetic Relationships among populations of Gibberella zeae from barley, wheat, potato, and sugar beet in the Upper Midwest of the United States. Phytopathology, accepted on 2 June Abstract: Burlakoti, R.R., Ali, S., Secor, G.A., Neate, S.M., McMullen, M., and Adhikari, T Impact of cereal and non-cereal hosts on trichothecene producing genotypes of Fusarium graminearum. Phytopathology 97:S15. Proceedings: McMullen, M Integration - The key to managing Fusarium head blight (FHB = scab) in wheat. Pages in: Proc Illinois Crop Protection Technology Conf., Jan. 3-4, Univ. of Illinois, Champaign, IL. McMullen, M Integrated strategies for FHB management, a Northern Great Plains perspective. Pages in: Proc. 5 th Canadian Workshop on Fusarium head blight. Winnipeg, Manitoba, Nov , Canadian Grain Commission, Winnipeg, Manitoba. McMullen, M Experiences in reducing disease and DON through components of FHB management. Pages in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. McMullen, M., Jordahl, J., and Meyer, S Differential effects of infection timing on Fusarium head blight and on DON and DON derivatives in three spring grains. Page 100 in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. (Form FPR07) 7

231 PI: McMullen, Marcia USDA-ARS Agreement #: McMullen, M., Meyer, S., and Jordahl, J Effects of fungicide timing on Fusarium head blight and on DON and DON derivatives in three spring grains. Pages 101 in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. Paul, P., Madden, L., McMullen, M., Hershman, D., Sweets, L., Wegulo, S., Bockus, W., Halley, S., and Ruden, K An integrated approach to managing FHB and DON in wheat: Uniform trials Pages in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. Paul, P., Lipps, P., Hershman, D., McMullen, M., Draper, M., and Madden, L A quantitative synthesis of the relative efficacy of triazole-based fungicides for FHB and DON control in wheat. Pages in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. Paul, P., Madden, L., McMullen, M., Hershman, D., Brown-Rytlewski, D., Sweets, Adee, E., Bradley, C., Padgett, B., and Ruden, K Fungicide effects on FHB and DON in wheat across multiple locations and wheat classes: Uniform fungicide trials Pages in: Proc National Fusarium Head Blight Forum. Dec. 2-4, 2007, Kansas City, Kansas. US Wheat and Barley Scab Initiative, Michigan State Univ., East Lansing, MI. Extension: McKay, K., et al Winter wheat variety by fungicide trial, Berthold, ND. Page 573 in: Crop Production Guide 2008, No. 18. NDSU Extension Service Publication. Fargo, ND. 575 p. McKay, K., et al Winter wheat variety by fungicide trial, Roseglen, ND. Page 574 in: Crop Production Guide 2008, No. 18. NDSU Extension Service Publication, Fargo, ND. 575 p. McMullen, M., et al North Dakota State University Small Grain Fungicide Field Trials compilation. NDSU Dept. of Plant Pathology, Fargo, ND. 82 p. Found on-line at: (Form FPR07) 8

232 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Mohamed Mergoum Institution: North Dakota State University Address: Department of Plant Sciences Rm 166 Loftsgard Hall Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 117,631 Mapping and Pyramiding Resistance Genes and Developing Hard Red Spring Wheats Resistant to FHB. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Development of Hard Red Spring Wheat Cultivars Resistant Scab $117,631 Disease. Total Award Amount $ 117,631 06/17/2008 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

233 PI: Mergoum, Mohamed USDA-ARS Agreement #: Project 1: Development of Hard Red Spring Wheat Cultivars Resistant Scab Disease. 1. What major problem or issue is being resolved and how are you resolving it? Scab or Fusarium head bight (FHB) remains a major threat to wheat production and industry in the Northern Central Plains of the US. It is a complex disease that reduces significantly the grain yield and impacts negatively the wheat quality. In ND, the disease had tremendous implications, particularly on HRSW producers; users; and export market. This problem is being addressed by the NDSU HRSW wheat breeding program. Our goal is to resolve this problem by the development of elite and adapted genotypes/ lines/cultivars and breeding populations that incorporate diverse genetic resistance to the disease with desired agronomic and quality traits. Our strategy is based on pyramiding several types of genetic resistance to FHB from diverse sources into adapted cultivars using classical breeding methods and appropriate novel technologies such as selected molecular markers. We believe that genetic resistance provides a strategic long-term, economically, and environmentally sound solution to the problem. During the growing cycle, our efforts have continued to develop elite HRSW germplasm and cultivars that are adapted to ND in particular, and spring wheat region, in general. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Our main accomplishments are illustrated in the release of many HRSW cultivars and germplasm that are hallmark of wheat production in the spring wheat region in particular and in the US as a whole. These varieties are grown on large acreages in ND and neighboring states generating millions of dollars of benefits for the growers, the industry and export market. These cultivars are the following: The most recent release by our HRSW program (2007) is Faller. Before its release, Faller was tested under ND 805 experimental line for many years in different nurseries in our breeding program and in the regional nurseries, including URN and URSN. Faller is a derivative cultivar of Sumai3. Hence it has medium resistance to FHB, similar to Alsen, a very popular HRSW cultivar in the spring region. However, Faller has very high yield and very good leaf disease package compared to Alsen. Faller is recommended for high rainfall and FHB prone spring wheat regions. Prior to the release of Faller, Howard (ND 800) HRSW wheat variety was released in 2006 with good FHB resistance level comparable to Steele-ND. Both Howard and Steele- ND have FHB resistance different from Alsen (Sumai3). Howard has wide adaptation and recommended mainly, for the regions where Reeder and Alsen are grown because of its high yield level and good leaf diseases resistances package. Howard main attributes are FHB resistance, high yield and quality, and excellent leaf disease package compared to Alsen and Reeder. Glenn (ND747) released in 2005 is now The LEADING HRSW cultivar in the spring wheat region replacing Alsen that dominated the wheat production for the last 5 years. Glenn with parentages involving both Sumai-3 and Steele-ND has higher FHB resistance (Form FPR07) 2

234 PI: Mergoum, Mohamed USDA-ARS Agreement #: level (better than Alsen variety). In addition Glenn has excellent quality attributes allowing it to be the quality standard by the Wheat Quality Council and the industry in the USA. Based on the US-Wheat Associates survey (OVA), Glenn is now the most preferred cultivar by the wheat importers worldwide. Glenn is expected to dominate the wheat production for some years. The 2004 HRSW release, Steele-ND (ND 741) has good FHB resistance level and wide adaptation is now a major grown cultivar in the spring wheat region. Steele-ND has been grown on more than 9% of ND acreages in the past two years. It is recommended mainly, for the regions where Reeder and Alsen are grown because of many attributes including high yield level. It expected that Steele-ND will replace significant acreages of Alsen and Reeder across all the State and probably other cultivars in the region. In 2007, we have released ND 756 as a germplasm with FHB resistance (see publications). Prior to that, we have released several key sources of FHB resistance wheat germplasm including, ND 2710, ND 744, and ND 751 (See Crop Sciences). Impact: In the past 5 years, more than 50% (3.5-4 million acres) of ND spring wheat have been grown to NDSU cultivars. Among these common grown NDSU cultivars, Alsen, Steele- ND and Glenn HRSW cultivars have resistance to FHB and excellent agronomic/quality traits contribute. These three (3) cultivars have occupied more than 45.1% (2.85 million acres) of wheat grown in ND. These figures show that the impact on wheat business (growers, industry and export market) of the FHB resistant HRSW cultivars developed by this program using partly, the USWBSI initiative funds is phenomenal. In addition, the NDSU HRSW cultivars are also grown in neighboring states (MN, SD, and MT) where spring wheat is a major crop and FHB is a threat. This is an important impact that should be factored in as well. For example, Reeder, the 1999 NDSU release is the leading wheat cultivar in MT 26%. Releasing superior and very high quality HRSW cultivars with improved FHB resistance has allowed ND growers to be competitive in the wheat market at the national and international levels. The HRSW germplasm with FHB resistance that we have released is well known and extensively used in the breeding program nationally and worldwide. Our HRSW breeding program is now a Center of excellence for wheat germplasm with high quality and good sources of FHB resistance. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The above accomplishments/results (scab resistance HRSW cultivars and germplasm) by the NDSU HRSW breeding program has allowed the US wheat growers in the spring region to grow once again, on a large scale (3-4 million acres), wheat in scab prone regions. This obviously, has been generating for the growers, hundreds of millions of dollars annually. Similarly, the wheat industry is having better wheat quality for their products, particularly (Form FPR07) 1

235 PI: Mergoum, Mohamed USDA-ARS Agreement #: during the scabby years. In addition, as more than 50% of total HRSW is exported worldwide for its superior quality, the NDSU HRSW cultivars with high quality and scab resistance has allowed our export market to be more competitive at the international level. Hence, substantial financial impact on wheat industry and export market due to NDSU HRSW cultivars has been generated. Finally, NDSU germplasm with FHB resistance and high quality is being used nationally and internationally in the research improvement program as a hallmark sources for these traits. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. Mergoum, M., R. C. Frohberg, and R. W. Stack Registration of spring wheat germplasm ND 756 combining resistances to Fusarium head blight, leaf spotting, and rusts diseases. Journal of Plant Registration Vol. 2, No.1: Zhang, G., and M. Mergoum Molecular mapping of kernel shattering and its association with Fusarium head blight resistance in a Sumai3 derived population. Theoretical and Applied Genetics 115: Zhang, G., and M. Mergoum Developing evaluation methods for kernel shattering in spring wheat. Crop Science 47: Mergoum, M., R. C. Frohberg, and R. W. Stack Registration of spring wheat germplasm ND 751 resistant to Fusarium head blight leaf and stem rusts. Crop Science 47: (Form FPR07) 2

236 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Eugene Milus Institution: University of Arkansas Address: Department of Plant Pathology 217 Plant Science Bldg. Fayetteville, AR Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Management of FHB in Arkansas. Title: FY07 ARS Award Amount: $ 71,881 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount VDUN Developing FHB-Resistant Wheat Cultivars for the Midsouth. $71,881 Total Award Amount $ 71,881 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

237 PI: Milus, Eugene USDA-ARS Agreement #: Project 1: Developing FHB-Resistant Wheat Cultivars for the Midsouth. 1. What major problem or issue is being resolved and how are you resolving it? The major problems are that wheat cultivars currently grown in the region have little or no resistance to FHB, levels of DON in harvested grain sometimes exceed the allowable level, and a nivalenol chemotype of the FHB pathogen has been found recently at high frequency in the region. The Arkansas program is resolving this by developing several wheat cultivars and lines with more competitive yield potential and moderate to high levels of resistance to FHB and other diseases that are important in the region. The Arkansas program also assisted other breeding programs by evaluating the Southern Winter Wheat Scab Nursery and lines from the LSU wheat breeding program for resistance to FHB and other diseases. A Ph. D. graduate student is conducting research to 1) determine if wheat lines with resistance to the DON chemotype also will be resistant to the nivalenol chemotype, 2) to quantify resistance to DON injected into florets and determine the utility of this variable for evaluating resistance to FHB, 3) to identify wheat lines with resistance to initial infection (type I resistance) and to DON and nivalenol accumulation in harvested grain, and 4) to improve the methodology for evaluating resistance to initial infection and mycotoxin accumulation in grain. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The most important accomplishment has been the development of three lines with good FHB resistance. The resistance was verified in the 2007 Southern Uniform Winter Wheat Scab Nursery. Three of the Arkansas entries, AR , AR , and AR850-1 were ranked in the top ten for FHB index across all locations at 15, 16, and 16, respectively. This was not significantly different than either of the resistant checks, Ernie and Bess at 12 and 7 respectively. They were also low in DON (9.1, 9.0, and 13.3 ppm) compared to the checks (9.1 and 5.3 ppm). Lines from the 2008 Southern Winter Wheat Scab Nursery, Arkansas and LSU wheat breeding programs, and Arkansas germplasm enhancement program were evaluated in inoculated, misted nurseries at two field locations to identify resistant lines. Lines from the 2008 Southern Winter Wheat Scab Nursery also were evaluated for type I resistance in the greenhouse. Fifteen winter wheat lines with diverse sources of FHB resistance ranked similarly for resistance to DON and NIV chemotypes of the pathogen in evaluations for type I and type II resistance, indicating that the resistance genes currently used in US soft red winter wheat programs are effective against both chemotypes. Experiments with injection of purified DON into wheat florets confirmed that resistance gene FHB1 confers resistance to DON and determined that this is likely the only FHB resistance gene that confers both resistance to DON and resistance to spread within a spike. The level of type I resistance in 15 winter wheat lines was quantified. Efforts to quantify resistance to DON accumulation in grain and to improve methodology for evaluating resistance to initial infection and mycotoxin accumulation in grain are still in progress. (Form FPR07) 2

238 PI: Milus, Eugene USDA-ARS Agreement #: Impact: Two of the three lines are either commercially available or are being closely looked at for release. AR850-1 was released and licensed to a commercial seed company in Arkansas. AR was entered in the 2008 Uniform Eastern Soft Wheat Nursery as well as the Arkansas State Variety Trials. The ultimate goal of the project is to get FHB-resistant cultivars into growers fields. AR will also give other breeders the chance to use an FHBresistant line with good stripe rust and winterhardiness (even for the Midwest) as a parent in their program. Evaluation of breeding lines for resistance to FHB and other diseases facilitates the development of FHB-resistant cultivars. Cultivars with resistance to DON chemotypes also will be resistant to NIV chemotypes, therefore a special breeding program for resistance to NIV chemotypes is not necessary. Resistance gene FHB1 was shown to be the only gene that confers both resistance to DON and resistance to spread within a spike, indicating that this gene is more valuable than previously believed because it has a unique mode of action. Lines with high levels of type I resistance were identified, indicating that suitable sources of resistance to initial infection are available in adapted lines. Improving the methodology for evaluating resistances to initial infection and mycotoxin accumulation in grain will facilitate development of resistant cultivars. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Growers will have the opportunity to plant AR850-1, and the last set of evaluations for AR will determine if it becomes commercialized. The breeding community will also have adapted germplasm to utilize with AR being a southern line that derives resistance from a more northerly background. Evaluation of breeding lines for resistance to FHB and other diseases provides data that breeders use to make decisions relative to advancement of lines and release of cultivars. Breeders will be confident that cultivars developed for resistance to DON chemotypes also will be resistant to NIV chemotypes. Breeders have access to adapted lines with high levels of type I resistance. The scientific community has new knowledge that gene FHB1 has a unique mode of action. (Form FPR07) 3

239 PI: Milus, Eugene USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Horevaj, P., Milus, E. A., Gale, L. R., and Kistler, H. C Resistance in soft red winter wheat lines to deoxynivalenol and nivalenol chemotypes of Fusarium graminearum. Phytopathology 97S:48. Horevaj, P., and Milus, E. A Resistance of winter wheat to Fusarium head blight and mycotoxin accumulation. Pages in: Arkansas Crop Protection Association Research Abstracts Vol. 11. (Peter earned second place in the Ph. D. oral presentation competition.) Horevaj, P., and Milus, E. A Resistance of winter wheat to Fusarium head blight and mycotoxin accumulation. Gamma Sigma Delta Arkansas Chapter Student Research Competition, February 25, (Peter earned second place in the Ph. D. oral presentation competition.) Horevaj, P., Milus, E. A., Gale, L. R., and Kistler, H. C Resistance of winter wheat lines to deoxynivalenol and nivalenol chemotypes of Fusarium graminearum. Pages in: Proceedings of the 2007 National Fusarium Head Blight Forum. Milus, E. A., and Hedge, J Evaluations for FHB severity, Type II resistance, and stripe rust resistance. Pages 8, 21, and 26, respectively, in: 2007 Southern Winter Wheat Scab Nursery Report. J.P. Murphy and R.A. Navarro, editors. (Form FPR07) 4

240 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Michelle Mostrom Institution: North Dakota State University Address: Dept. of Veterinary and Microbiological Sciences P.O. Box Centennial Blvd. Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Diagnostic Services for Vomitoxin (DON) in Wheat. Title: FY07 ARS Award Amount: $ 71,564 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount FSTU-S Diagnostic Services for Vomitoxin (DON) in Wheat. $71,564 Total Award Amount $ 71,564 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

241 PI: Mostrom, Michelle USDA-ARS Agreement #: Project 1: Diagnostic Services for Vomitoxin (DON) in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium Head Blight (FHB) or scab can be a costly disease of cereal crops. Under environmental conditions of excessive moisture and alternating warm/cool temperature, Fusarium mold can grow in field crops and produce mycotoxins or secondary metabolites, including deoxynivalenol (DON) or vomitoxin. DON concentrations in wheat and barley can exceed recommended guidelines for grain use and render the cereals as unacceptable for processing into foods or animal feeds. The focus of the U.S. Wheat and Barley Scab Initiative (USWBSI) is to reduce FHB in wheat and barley. This grant provided DON analyses of samples for research scientists involved in projects for the USWBSI. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The USWBSI research scientists directly benefit from analytical identification and quantitation of DON in their research cereal samples. Knowledge of mycotoxins, specifically vomitoxin, in research plants or cereal grains yields important data for controlling Fusarium growth and mycotoxin production in specific research projects that will ultimately benefit the plant producers and consumers. The laboratory completed vomitoxin analyses on ~ 10,100 ground feed samples. The samples were submitted from 22scientists in seven states, including North and South Dakota, Nebraska, Kansas, Missouri, Virginia, and Montana. The laboratory also provided multiple Fusarium mycotoxin screen on ~50 grain samples and used the screen for quality assessment on control pool cereal samples. The laboratory participated in the check sample system to compare analytical data from vomitoxin analyses with laboratories in Michigan, North Dakota, and Minnesota. Impact: Vomitoxin or DON analysis in wheat samples is a basic service to USWBSI plant scientists and provides data used for evaluating mitigation methods for Fusarium head blight. This information is transferred to plant breeders and producers for cultivation of cereals grains that are potentially more resistant to Fusarium. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Knowledge of DON production in wheat research of plant scientists is important for evaluation of methods aimed at mitigating FHB in cereal crops. This is a basic component for USWBSI research providing scientists, breeders and producers with end-result data on the effectiveness of reducing FHB in cereal cultivars. (Form FPR07) 2

242 PI: Mostrom, Michelle USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. No direct publications by the PI, but data used by other USWBSI researchers (Form FPR07) 3

243 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Gary Muehlbauer Institution: University of Minnesota Address: Department of Agronomy and Plant Genetics 411 Borlaug Hall 1991 Upper Buford Circle St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Molecular Genetic Approaches to Develop Scab Resistance. Title: FY07 ARS Award Amount: $ 100,523 USWBSI Individual Project(s) USWBSI Research Area * GET HGG ARS Adjusted Award Project Title Amount Developing and Characterizing Transgenic Wheat for Scab Resistance. $41,986 Mechanisms and Essential Genes for Resistance to Fusarium Head Blight. $ 58,537 Total Award Amount $ 100,523 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

244 PI: Muehlbauer, Gary USDA-ARS Agreement #: Project 1: Developing and Characterizing Transgenic Wheat for Scab Resistance. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB or scab) is a disease that can devastate wheat. To enhance FHB resistance in wheat, we are developing transgenic wheat lines with potential resistance genes and testing these lines for scab resistance. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: 1) We identified two wheat lines carrying a chitinase transgene that exhibited enhanced resistance in the field. A paper was published in the Journal of Experimental Botany describing the chitinase lines. 2) We also identified wheat transgenic lines carrying a ribosomal inhibitor protein (RIP), ß-1,3- glucanase/tlp, and RIP/tlp transgenes that exhibited enhanced resistance in the field. 3) We identified transgenic wheat lines carrying a glutathione-s-transferase, a lipid transfer protein, a JAMyb transcription factor, and a germin-like protein that exhibited enhanced resistance in the greenhouse. Impact: We identified transgenic wheat lines that exhibited enhanced resistance in the greenhouse and field. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community now has access to transgenic wheat lines that exhibit reduced FHB severity. These lines provide another source of variation for FHB resistance. In addition, the scientific community now has new information on the functional impact of a variety of genes on FHB resistance. (Form FPR07) 2

245 PI: Muehlbauer, Gary USDA-ARS Agreement #: Project 2: Mechanisms and Essential Genes for Resistance to Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB or scab) is a disease that can devastate the small grains wheat and barley. The wheat and barley transformation efforts have a limited number of genes that have the potential to reduce FHB. In addition, there is a limited amount of information pertaining to the molecular genetic interaction between the small grains and Fusarium graminearum. Our goal is to identify the mechanisms and essential genes for wheat and barley scab resistance. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishments: 1) Described the impact of deoxynivalenol treatment on barley. We identified 135 barley genes that respond specifically to DON treatment. These genes encode proteins involved in trichothecene detoxification and transporter function, cell death and ubiquitination processes. A manuscript is in preparation describing this work that we plan to submit to Molecular Plant Pathology. All data will be released on the web at PLEXdb ( 2) To conduct functional tests of genes potentially providing resistance, we established virusinduced gene silencing (VIGS) assays in my lab. We used VIGS to test a variety of potential FHB resistance genes that we identified in our gene expression (GeneChip) experiments. We tested four different transcription factors that have been identified in our GeneChip experiments and are involved in other plant-pathogen interactions. Our first experiments have been completed and we are evaluating the results and repeating the experiments. We also developed VIGS constructs for three other genes. 3) Reexamined data from our GeneChip experiments focused on a near-isogenic line pairs carrying resistant and susceptible alleles at FHB resistance QTL in barley and wheat. A manuscript will be submitted in July to Molecular Plant-Microbe Interactions describing the work from the wheat near-isogenic line. All data will be released on the web at PLEXdb ( 4) Prepared BAC libraries for F. graminearum for physical mapping the genome and genome sequencing. Papers in Genome and Science were published using the BAC libraries. 5) My laboratory collaborated with Dr. Kevin Smith (University of Minnesota) to fine map a barley QTL region associated with FHB resistance, and identified a useful line and associated markers for barley breeding. A paper was published on this topic in Theoretical and Applied Genetics. Impact: (Accomplishment 1) Identified barley genes that respond to DON treatment. These genes potentially control a variety of cellular processes including: trichothecene detoxification and transport, cell death and ubiquitination processes. (Accomplishments 2) We established VIGS in my laboratory to test gene function. We began to use VIGS to test the impact of four genes on FHB resistance. (Accomplishment 3) We are identifying genes that differentiate resistant from susceptible interactions, and may be involved in providing resistance. (Form FPR07) 3

246 PI: Muehlbauer, Gary USDA-ARS Agreement #: (Accomplishment 4) The physical map and genome sequence of F. graminearum is an important resource for future studies of the fungus. (Accomplishment 5) My laboratory was involved with developing markers and a genetic line that is being used in the University of Minnesota barley-breeding program. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community has access to an unprecedented amount of expression data that defines the barley-f. graminearum interaction, the barley-trichothecene interaction and the wheat-f. graminearum interaction. In addition, candidate genes have been identified for involvement in FHB resistance. The barley line carrying the FHB resistance QTL and the development of the associated markers are useful for barley breeding. The F. graminearum physical map and genome sequence provided an important resource for future understanding of the fungus. (Form FPR07) 4

247 PI: Muehlbauer, Gary USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Refereed Publications: Chang, Y-L., S. Cho, H.C. Kistler, H-C. Sheng and G.J. Muehlbauer Bacterial artificial chromosome-based physical map of Gibberella zeae (Fusarium graminearum). Genome 50: Nduulu, L.M., A. Mesfin, G.J. Muehlbauer and K.P. Smith Analysis of the chromosome 2(2H) region of barley associated with the correlated traits Fusarium head blight resistance and heading date. Theor. Appl. Genet. 115: Boddu, J., S. Cho and G.J. Muehlbauer Transcriptome analysis of trichothecene-induced gene expression in barley. Mol. Plant-Microbe Interact. 20: Cuomo, C.A., U. Güldener, J-R. Xu, F. Trail, B.G. Turgeon, A. Di Pietro, J.D. Walton, L-J. Ma, S.E. Baker, M. Rep, G. Adam, J. Antoniw, T. Baldwin, S. Calvo, Y-L. Chang, D. DeCaprio, L.R. Gale, S. Gnerre, R.S. Goswami, K. Hammond-Kosack, L.J. Harris, K. Hilburn, J.C. Kennell, S. Kroken, J.K. Magnuson, G. Mannhaupt, E. Mauceli, H.-W. Mewes, R. Mitterbauer, G. Muehlbauer, M. Münsterkötter, D. Nelson, K. O Donnell, T. Ouellet, W. Qi, H. Quesneville, M.I.G. Roncero, K-Y. Seong, I.V. Tetko, M. Urban, C. Waalwijk, T.J. Ward, J. Yao, B.W. Birren, and H.C. Kistler The Fusarium graminearum genome reveals a link between localized polymorphism and pathogen specialization. Science 317: Shin, S., C.A. Mackintosh, J. Lewis, S.J. Heinen, L. Radmer, R. Dill-Macky, G.D. Baldridge, R.J. Zeyen and G.J. Muehlbauer Transgenic wheat expressing a barley class II chitinase gene has enhanced resistance to Fusarium Head Blight. J. Exp. Bot. 59: Jia, H., S. Cho and G.J. Muehlbauer. Transcriptome analysis of the wheat 3BS Fusarium head blight resistance QTL during Fusarium graminearum infection. In preparation for submission to Molecular Plant-Microbe Interactions. Gardiner, S., J. Boddu and G.J. Muehlbauer. Deoxynivalenol-induced gene expression in barley. In preparation for submission to Molecular Plant Pathology. Abstracts: Jia, H., S. Cho and G.J. Muehlbauer Transcriptome analysis of wheat near-isogenic line pair carrying fusarium head blight resistant and susceptible alleles. Plant and Animal Genome Meeting Abstracts. Gardiner, S.A., H. Jia, J. Boddu and G. J. Muehlbauer Microarray analysis of deoxynivalenol-induced gene expression in susceptible (cv. Morex) barley. American Phytopathological Society Meeting Abstracts Shin, S.H., J.M. Lewis, C.A. Mackintosh, A. Elakkad, K. Wennberg, S.J. Heinen, R. Dill-Macky, G. J. Muehlbauer Transgenic wheat with enhanced resistance to Fusarium Head Blight. National Scab Forum Abstracts. Boddu, J., H.C. Kistler and G.J. Muehlbauer Role of trichothecenes in the barley- Fusarium graminearum interaction. National Scab Forum Abstracts. (Form FPR07) 5

248 PI: Muehlbauer, Gary USDA-ARS Agreement #: Jia, H., B.J. Steffenson, and G.J. Muehlbauer Effects of agronomic and morphological characters on FHB severity, deoxynivalenol and ergosterol concentration in near-isogenic line pairs of barley. National Scab Forum Abstracts. Oral Presentations: The challenge for barley genomics: integrating phenotypes and genotypes at BASF Plant Science LLC, Research Triangle Park, NC Genomics of the Triticeae-Fusarium graminearum interaction at the Danforth Center, St. Louis, MO (Form FPR07) 6

249 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: J. Paul Murphy Institution: North Carolina State University Address: Department of Crop Science Box 7629 Raleigh, NC Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 54,279 Enhancement of Fusarium Head Blight Resistance in the Southeastern U.S. Germplasm Pool. USWBSI Individual Project(s) USWBSI Research Area * HGR VDUN ARS Adjusted Award Project Title Amount Introgression of FHB Resistance from Hexaploid and Intergeneric Hybrids into Soft Red Winter Wheat. $15,434. Development of Fusarium Head Blight Resistant Wheat for the Southeastern United States. $ 38,845 Total Award Amount $ 54,279 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

250 PI: Murphy, J. Paul USDA-ARS Agreement #: Project 1: Introgression of FHB Resistance from Hexaploid and Intergeneric Hybrids into Soft Red Winter Wheat. 1. What major problem or issue is being resolved and how are you resolving it? We are seeking to provide eastern U.S. wheat breeders with novel sources of resistance to Fusarium Head Blight in adapted backgrounds. Donor sources include exotic cultivars, a diploid wheat relative, and intergeneric (Triticum aestivum x Lophopyrum elongatum) hybrids from the Sando collection. Donor sources are backcrossed to susceptible North Carolina adapted germplasm. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: 1. Exotic Cultivars: Greenhouse An average of 12 BC 2 F 1 progeny in each of 12 backcross families were evaluated by single floret inoculations, and an average of 8 progeny per family were selfed to produce BC 2 F 2 seed. The donor parents (Argentina 8, Argentina 93, Brazil 5, CIMMYT 21, CIMMYT 26, CIMMYT 34, and PI ) were identified as having resistance distinct from Sumai 3 by Liu and Anderson (Crop Sci., 43: ) based on allelic content at five SSR loci. Field Approximately 300 BC 1 F 2:3 lines involving donor parents (PI , PI , PI , PI , PI 81791, PI and PI ) identified as having resistance distinct from Sumai 3 by Liu and Anderson (Crop Sci., 43: ) were evaluated in an inoculated and misted nursery at Kinston NC in The recurrent parent was a susceptible soft red winter wheat adapted to the southeastern US. Resistant heads were tagged and harvested separately. BC 2 F 2 materials involving the same donor and recurrent parents were advanced in bulk in the same nursery. 2. T. monococcum: Field Two hundred sixty BC 2 F 4:5 lines from a backcross involving a T.monococcum donor (PI ) and the susceptible NC were evaluated in a misted and inoculated head row nursery. Nine lines displaying better resistance than the recurrent parent were harvested in bulk. Unfortunately, the level of resistance in the selected lines was little better than the recurrent parent. 3 Sando Intergeneric Hybrids: Field Two hundred thirty BC 1 F 2:3, BC 1 F 3:4, and BC 2 F 2:3 progeny in each of four backcross families (Form FPR07) 2

251 PI: Murphy, J. Paul USDA-ARS Agreement #: were evaluated in an inoculated and misted nursery. The donor parents (PI , PI , PI , and PI ) were previously identified as having excellent Type II resistance. Heads exhibiting low FHB symptoms were tagged and harvested. Impact: Because the best known sources of resistance to Fusarium head blight in the cultivated wheat gene pool are partial rather than complete, it is timely to seek additional resistance sources to complement those genes already identified. We believe we have moved potentially novel sources of resistance from exotic cultivated wheat and, wheatgrass into southeastern U.S. adapted soft red winter wheat. However additional evaluations will be necessary to confirm the presence of the novel resistance genes. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Within two seasons we should enter some of these these materials in uniform cooperative nurseries so breeders in the public and private sectors can evaluate the germplasm and have unfettered access to a greater genetic diversity of resistance genes to incorporate into their own variety development programs. (Form FPR07) 3

252 PI: Murphy, J. Paul USDA-ARS Agreement #: Project 2: Development of Fusarium Head Blight Resistant Wheat for the Southeastern United States. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium Head Blight is a major concern to the wheat industry in North Carolina due to the annual occurrence of the disease in commercial production. We are resolving this problem by developing southeastern adapted cultivars and germplasms with moderate to high levels of FHB resistance combined with superior productivity, disease and insect resistance and enduse quality demanded by the industry. To provide breeding programs with reliable, independent data on FHB resistance in advanced generation lines the Uniform Southern Soft Red Winter Wheat Scab Nursery is coordinated by North Carolina State University. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Advanced Lines Field data collected under unusually severe natural and inoculated FHB epidemics combined with Type II greenhouse data provided an opportunity to observe program progress under a worst-case scenario. Two advanced lines were identified for release in 2009 (NC ) and 2010 (NC ) with FHB resistance at least as good as the moderately resistant NC- Neuse. Both genotypes have native resistance of unknown origin. Both have excellent overall agronomic and end-use quality characteristics. Eleven of 49 entries in the 2008 Advanced Test had field resistance at least as good as the moderately resistant NC-Neuse. Ten lines had native sources of resistance, and one contained Fhb1. The yield of the latter genotype was not significantly different from the highest yielding entry. Twenty of 54 entries in the 2008 Preliminary Test had field resistance at least as good as the moderately resistant NC-Neuse. Nineteen of the 56 entries in the 2009 Preliminary Test will contain exotic (CIMMYT, Chinese, Italian, Japanese) FHB resistant parents in their pedigrees. Only six of the remaining 37 entries have no FHB resistant parents in their pedigrees. Seventeen of the 30 entries in the 2009 Advanced Test will contain FHB resistant parents in their pedigrees. One entry contains Fhb1. A second year of data collected in both natural and inoculated FHB nurseries on 50 entries in the NC Official Variety Test was obtained and made available to producers and end-users. The Uniform Southern Nursery program contained 45 entries submitted by eight US public and private breeding programs, plus one Romanian program. Eleven cooperators (9 US and one each in Romania and Hungary) returned laboratory, greenhouse and/or field nursery data. The data were summarized in a nursery report distributed to wheat breeders and pathologists in Dec The Uniform Southern Nursery containing 52 entries submitted by nine US public and private breeding programs was distributed for planting in fall (Form FPR07) 4

253 PI: Murphy, J. Paul USDA-ARS Agreement #: Early Generation Materials Approximately 250 crosses involving FHB resistant parents were made during winter Twenty three 3-way F 1 populations and two F 2 populations were enhanced for Fhb1 frequency using MAS. A MAS project to introgress the Qfhs.ndsu-3AS resistance into the NC-Neuse background was successfully completed with the derivation of BC 2 F 2:3 lines homozygous for the resistance gene. A similar effort to introgress the 3A and 5A regions controlling Type 1 resistance in Frontana into a susceptible southeastern background resulted in the distribution of BC 2 F 2 seed to five public programs for further selection. The BC 2 F 2 seed was harvested from BC 2 F 1 plants heterozygous for the Frontana alleles. Three F 1 populations with parentage containing Fhb1 in a semi-adapted background and highly productive, adapted lines are being readied for corn pollination and doubled haploid regeneration. Impact: The Uniform Southern Nursery provided cooperators with comprehensive evaluations of the FHB resistance in their advanced generation lines. These data are vital for release decisions and for choosing and sharing parents for further population development. Our sharing of Southeastern adapted germplasm with the novel resistance gene Qfhs.ndsu- 3AS broadens the genetic variation available in a desirable genetic background. The NC program produced advanced generation lines that have moderate to good levels of FHB resistance combined with good overall agronomic and end-use quality. Several of these are of sufficient overall merit to enter regional cooperative trials, which suggests they may be of variety quality. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Two lines are being increased for release in 2009 and 2010 that combine overall agronomic and end-use excellence with enhanced FHB resistance. Providing growers with more varietal choices with FHB resistance will lead to a decline in the use of susceptible cultivars that are high performers in the absence of FHB epidemics, but extremely poor performers in the presence of FHB epidemics. Uniform Nurseries provided rapid dissemination of resistant lines throughout the breeding community and provide a ready source of information on the true levels of resistance in advanced generation breeding lines. (Form FPR07) 5

254 FY06 (approx. May 06 April 07) PI: Murphy, J. Paul USDA-ARS Agreement #: FY06 Final Performance Report Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Murphy, J. P., and R. A. Navarro The Southern Uniform Winter Wheat Scab Nursery. p In S. M. Canty, A. Clark, D. Ellis and D. A. Van Sanford (eds.) Proc. of the 2007 National Fusarium Head Blight Forum, 2-4 Dec.; The Westin Crown Center, Kansas City, Missouri, USA. Univ. of Kentucky, Lexington, KY. Reports: Murphy, J. P. and R. A. Navarro Southern Uniform Winter Wheat Scab Nursery. Dept. Crop Science, N.C. State Univ., Raleigh. Also available at (Form FPR07) 6

255 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Stephen Neate Institution: North Dakota State University Address: Department of Plant Pathology 353 Walster Hall Box 5051 Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Management and Resistance Sources for Control of FHB in Barley. Title: FY07 ARS Award Amount: $ 62,635 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Preharvest Management Strategies in Barley to Reduce FHB and DON. $22,639 HGR Accelerated Screening of Hordeum Germplasm for FHB Resistance. $ 28,216 VDUN Screening Barley Lines for Scab Resistance in Cross-State and Region Uniform Nurseries. $ 11,780 Total Award Amount $ 62,635 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

256 PI: Neate, Stephen USDA-ARS Agreement #: Project 1: Preharvest Management Strategies in Barley to Reduce FHB and DON. 1. What major problem or issue is being resolved and how are you resolving it? Weather conditions in North Dakota during barley harvest slow crop maturity and can result in non-uniform crop maturity within a field. In most years barley producers choose to use windrowing or pre-harvest herbicides as desiccants to accelerate crop maturity and drying and allow clean threshing. Windrowing involves cutting the crop near ground level and then pushing it up into rows 3-4 ft wide and 1-2 ft high. Current pesticide registration in North Dakota permits preharvest application of 2,4-D ester, metsulfuron and glyphosate. In addition paraquat and dicamba have been used experimentally. In contrast to wheat, barley is susceptible to infection by Fusarium from head emergence through to harvest. Windrowing and pre-harvest herbicide usage can significantly influence the effectiveness of the best fungicides, resistance and management practices for control of FHB if infection occurs late after fungicide effectiveness is reduced and when active plant resistance has ceased. In addition, these late season influences on FHB and DON levels in the grain can complicate the interpretation of predictive disease models which rely on climatic data collected during the flowering and grain ripening period. To provide recommendations for preharvest management strategies both plus and minus windrowing and 14 desiccant herbicide treatments were applied to both 2-rowed and 6- rowed barley. Grain was assessed for number of visually infected kernels immediately before swathing. Treatments were then swathed and standing barley. Swathing treatments followed commercial practice. Irrigation schedules to simulate rainfall were applied at regular intervals after the crop was windrowed. After 2 weeks the grain was harvested and assessed for FHB incidence and severity as well as DON. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: In 2007 we demonstrated that while simulated rainfall increased both the incidence and severity of FHB in both Robust and Conlon there was no significant effect of swathing versus harvesting of the standing crop. A similar result was seen for DON accumulation, simulated rainfall increased DON in the crop but there was no significant effect of swathing versus harvesting of the standing crop on DON accumulation. As expected due to the stage of crop maturity when the simulated rainfall was applied, extra water had no effect on yield. In 2007, flooding of the trial site for a two week period destroyed the desiccant study and no data was able to be collected. Impact: (Form FPR07) 2

257 PI: Neate, Stephen USDA-ARS Agreement #: During the three years that we undertook this research we got one year in which swathing increased FHB or DON and two in which it did not, so it is difficult to give definitive information to farmers about the effect of preharvest swathing except to say that environmental conditions can influence the result of this treatment and farmers must weight the benefit of the treatment against potential for increased disease. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: An awareness that preharvest treatments in barley can in some years affect FHB incidence and severity as well as DON accumulation. (Form FPR07) 3

258 PI: Neate, Stephen USDA-ARS Agreement #: Project 2: Accelerated Screening of Hordeum Germplasm for FHB Resistance. 1. What major problem or issue is being resolved and how are you resolving it? Resistance to FHB in barley currently relies on only a few original sources of resistance and comprehensive screening of the US barley germplasm collection and the barley collection from the Dutch centre for genetic resources have found only few potentially new resistance sources. To broaden the genetic base of FHB resistance in barley, additional sources of resistance need to be identified and exploited as soon as possible. The project involves an accelerated screening in replicated experiments at several locations in North America, diverse Hordeum germplasm from as yet unscreened genebank collections around the world. To achieve this aim 550 lines were screened simultaneously in irrigated an inoculated nurseries at Fargo and Langdon ND. The two areas were chosen as they are climatically and geographically different. In addition this allows us to spread the load as Fargo ripens a week earlier than Langdon. Experimental units in ND nurseries consisted of single short rows and arranged in an augmented block design. Checks were Stander (FHB susceptible six-rowed cultivar), Chevron (FHB resistant six-rowed cultivar), Conlon (FHB susceptible two-rowed cultivar), and CIho 4196 (FHB resistant two-rowed accession) as well as IB1 and IB2. Checks were sown every 50 entries. Entries were inoculated using two applications of grain spawn, once before head emergence and the second prior to early dough. Fusarium head blight severity was determined at the mid dough stage by rating on a scale of 1-5, 20 spikes per row. Entries were scored for flowering date, height and other important agronomic characteristics. At maturity DON analysis was undertaken. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Seventy eight of the 550 lines at Langdon had a rating of 1 which was comparable to the resistant checks CI4196 and Chevron. The susceptible checks Stander and IB2 had mean ratings of 3-4. At Fargo 64 of the 550 had a rating of 0 or 1 which was comparable to the resistant checks CI4196 and Chevron. The susceptible checks Stander and IB2 had mean ratings of 2-3. Twenty three lines were selected as having low FHB symptoms and they were tested for DON accumulation. The 23 best lines had a mean DON of 6.3ppm with a range from ppm. The resistant check Chevron had a mean of 4.8 while CI4196 had a mean of 4.1. The susceptible check Stander had a mean of 22.1 while IB2 had a mean of Impact: It is clear that from this first round of testing that there are at least 20 lines that have both low FHB severity and low DON accumulation which approaches that found in the current resistant checks and these lines should now be tested in multiple field sites to ensure they (Form FPR07) 4

259 PI: Neate, Stephen USDA-ARS Agreement #: remain resistant under different environmental conditions and under a range of disease pressures. Once these lines are proven to be stable across environments they will be moved into the next stage of genotyping to ensure they are unique and crossed to breeding lines adapted to the upper Midwest. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We now have potentially 23 new sources of resistance to FHB and DON accumulation in barley. These are available breeders and pathologists for pre-breeding or breeding now or as more site years of testing become available. (Form FPR07) 5

260 PI: Neate, Stephen USDA-ARS Agreement #: Project 3: Screening Barley Lines for Scab Resistance in Cross-State and Region Uniform Nurseries. 1. What major problem or issue is being resolved and how are you resolving it? Regional uniform nurseries for crops are a standard method of comparison of advanced breeding lines and allow comparison of the relative advances made in the different breeding programs and foster germplasm exchange between diverse breeding programs. Within the nurseries advanced barley lines with putative FHB resistance were tested in mistirrigated sites as well as under rainfed conditions. Rainfed conditions represent those experienced by farmers in the regions in which the nursery is sown. Mist-irrigated nurseries that are artificially inoculated with Fusarium graminearum are needed so data can be collected in years when environmental conditions are not conducive for natural infection and to determine the stability of the putative resistance under high disease pressure. The series of FHB screening nurseries in this project has been carried out for more than 10 years and is currently known as the North American Scab Evaluation Nursery (NABSEN) nurseries. In 2007 the nurseries were grown at Fargo, Langdon, Osnabrock and Carrington, ND; St. Paul and Crookston MN; Brandon Canada and Hangzhou, China. This nursery includes breeding lines with putative FHB resistance from NDSU 2-rowed and 6-rowed, Minnesota State University, Busch-Ag and Agriculture & Agri-Food Canada barley breeding programs. FHB severity and DON accumulation are determined as well as agronomic characters such as heading date and environmental data such as rainfall. The objective of this project is to coordinate the screening in uniform FHB screening nurseries in North America of elite barley germplasm from breeding programs developing cultivars adapted to the upper Midwest barley growing region. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The 2007 North American Barley Scab Evaluation Nursery (NABSEN) was grown at Fargo, Langdon, Osnabrock and Casselton, ND; St. Paul and Crookston MN, Brandon, Manitoba; El Batan, Mexico and Hangzhou China. The nursery contained 54 lines including 6 resistant and susceptible controls. In Casselton the Busch Ag trial experienced seasonal conditions caused problems in the nursery, so that data is not available. All other sites experienced adequate to good conditions for the development of disease and the recording of relative levels of resistance in the different breeding programs. The resistance of the cultivars relative to the resistant and susceptible checks varied by location. Brandon had very high severities, Crookston was high, that remaining sites were (Form FPR07) 6

261 PI: Neate, Stephen USDA-ARS Agreement #: moderate to high except for El Batan which was low. For the ranking of lines, in general the inoculated irrigated sites or dryland sites with high rainfall had the highest correlations among themselves and low correlations with other dryland sites. Similarly, for the ranking of lines dryland sites had the highest correlations sites among themselves but low correlations with irrigated sites. When averaged over all sites, with 3 of the NDSU 6-rowed line and several of the the M and FEG lines from the University of Minnesota displaying the best resistance although all were short of Chevron the resistant 6-rowed check and CI4196 the resistant 2-rowed check. Similar trends were seen with DON accumulation. Environmental data was again added to this years NABSEN report so that breeders can better analyze performance and understand how environmental conditions influenced the disease present. Significant progress is being made toward developing FHB resistant barley cultivars. The project has allowed breeding programs with slower progress access to genetic material from the more advanced programs. Impact: All North American barley breeders have access to the data collected in this project. The information is made available electronically and in hard copy by the USWBSI meeting in December and even earlier by request. The breeders are able to use the relative performance data make decisions about continuing or dropping development of particular breeding lines. The project has also allowed breeding programs with slower progress to have access to genetic material from the more advanced programs or make decisions on the use of material being developed in foreign breeding programs such as the three Canadian programs and the CIMMYT/ICARDA program who submit entries to the nursery. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Breeders now have 1) tests of the resistance stability of their breeding lines across a range of environments and disease pressures, 2) a measure of the resistance in their advanced breeding material compared to those of the other barley breeders in North America and CIMMYT/ICARDA, and 3) access to unique germplasm with resistance to FHB and DON accumulation. (Form FPR07) 7

262 PI: Neate, Stephen USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Williston Research and Extension Centre Field day 12 July 2007, (5 talks each to 50 people) Carrington Research and Extension Centre Field day 19 July 2007, (5 talks each to 50 people) Langdon Research and Extension Centre Field day 20 July 2007, (3 talks each to 40 people) Franckowiak, J.D., Horsley, R.D., Neate, S.M. and Schwarz P.B. (2007) Registration of Rawson Barley. Journal of Plant Registrations 1: Burlakoti, R.R., Ali, S., Secor, G.A., S. M. Neate, S.M., M. McMullen, M. and Adhikari, T. (2007) Genetic relationships among populations of Fusarium graminearum from cereal and noncereal hosts. Phytopathology 97:S15. Burlakoti, R.R., Ali, S., Secor, G.A., S. M. Neate, S.M., M. McMullen, M. and Adhikari, T. (2007) Impact of cereal and non-cereal hosts on trichothecene producing genotypes of Fusarium graminearum. Phytopathology 97:S15. (Form FPR07) 8

263 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Herbert Ohm Institution: Purdue University Address: Department of Agronomy 915 West State Street West Lafayette, IN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Breeding Soft Winter Wheat with Multiple FHB Resistance. Title: FY07 ARS Award Amount: $ 113,964 USWBSI Individual Project(s) USWBSI Research Area * HGR VDUN ARS Adjusted Award Project Title Amount Introgression of Fusarium Head Blight Resistance of Thinopyrum into Wheat. $30,244 Improvement of Soft Winter Wheat for Fusarium Head Blight Resistance. $ 83,720 Total Award Amount $ 113,964 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

264 PI: Ohm, Herbert USDA-ARS Agreement #: Project 1: Introgression of Fusarium Head Blight Resistance of Thinopyrum into Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight is a serious and frequently recurring disease of wheat (Triticum aestivum) in Indiana and surrounding regions. Effective, but only partial resistance has been identified in certain germplasm lines of wheat. We have identified resistance in tall wheatgrass, (T. ponticum) that is related to wheat. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have introgressed a segment of chromosome 7E of tall wheatgrass that replaced the distal 1/3 of the long arm of wheat chromosome 7D and that has a FHB resistance QTL, Qfhs.pur-7EL that we have documented as having a strong resistance effect in wheat by itself, and that augments the resistance of Fhb1 and other FHB resistance genes/qtl. We have also developed a wheat line that has a shorter 7E segment, the distal 1/5 of the long arm of 7DS-7DL7EL, and have mapped Qfhs.pur-7EL to the distal region of the chromosome. Impact: No commercial impact yet. We are developing adapted soft winter wheat lines in which Qfhs.pur-7EL is combined with other FHB resistance QTL from wheat. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We project that we will have one or more adapted soft winter wheat lines with Qfhs.pur-7EL available for seeding in the field by USWBSI cooperators in fall (Form FPR07) 2

265 PI: Ohm, Herbert USDA-ARS Agreement #: Project 2: Improvement of Soft Winter Wheat for Fusarium Head Blight Resistance. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) is a serious and frequently occurring disease of wheat in Indiana and surrounding regions. Some soft winter wheat cultivars have been released that have partial FHB resistance, but yield drag in cultivars that have FHB resistance from unadapted parent lines from other areas in the world has been a limitation and there is need to develop cultivars with more effective resistance by combining multiple resistance genes/qtl. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: In 2007 we released cultivar INW0731 and in 2008 we released the cultivars INW0801 and INW0803 all three having partial FHB resistance. INW0731 especially, and more variably INW0801 and INW0803 have ranked first or in the upper 10% of entries in Indiana multilocation tests conducted by Purdue University and in regional tests conducted by Wheat Tech Services. INW0731 is especially adapted to drought conditions. INW0803, with short and strong straw performs well under high management conditions. INW0801 matures early and is very desirable for doublecropping with soybeans following wheat harvest in the same season. Impact: Acreage of INW0731 is rapidly increasing (indicated by high demand for basic seed). Impacts of INW0801 and INW0803 will be more clearly determined in As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These three cultivars provide additional opportunities for wheat producers in Indiana to overcome the significant limitation of FHB to wheat production and grain quality. (Form FPR07) 3

266 PI: Ohm, Herbert USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Shen, X., L. Kong, and H. Ohm Marker-assisted reduction of a chromosome segment of Thinopyrum ponticum carrying Fusarium head blight resistance. Purdue University Agriculture Research Programs, Journal paper no Crop Science (submitted). Ohm, H., J. Anderson, and G. Buechley Registration of INW0731. Journal of Plant Registrations (will submit in August 2008). Ohm, H., J. Anderson, and G. Buechley Registration of INW0801. Journal of Plant Registrations (will submit in August 2008). Ohm, H. and G. Buechley Registration of INW0803. Journal of Plant Registrations (will submit in August 2008). (Form FPR07) 4

267 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Boyd Padgett Institution: Louisiana State University Address: Macon Ridge Research Station 212 Macon Ridge Road Building A Winnsboro, LA bpadgett@agcenter.lsu.edu Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 4,878 Fungicide Efficacy Evaluations for Managing Scab in Louisiana Wheat. USWBSI Individual Project(s) USWBSI Research Area * CBCC ARS Adjusted Award Project Title Amount Fungicide Efficacy Evaluations for Managing Scab in Louisiana $4,878 Wheat. Total Award Amount $ 4,878 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

268 PI: Padgett, Boyd USDA-ARS Agreement #: Project 1: Fungicide Efficacy Evaluations for Managing Scab in Louisiana Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium Head Blight (FHB), Fusarium graminearum, can be a major threat to wheat producers in the United States. Disease epidemics are favored by periods of high humidity and moderate temperatures during flowering. These conditions are common along the Louisiana Gulf Coast. Furthermore, increased corn acreage and the use of minimum tillage practices in corn and wheat production systems have increased the risk to this disease in this state. This disease is managed in part by using genetic resistance and fungicides. However, no commercially available fungicides are highly effective against F. graminearum; therefore, a multi-state screening effort is ongoing to identify specific chemistries efficacious against this pathogen. The objective of this project is to evaluate fungicides for the management of FHB in Louisiana. Fungicides were evaluated in LSU AgCenter small field plot tests at the Macon Ridge Research Station (northeast) and the Rice Research Station (southwest). Each location represents a unique environment (e.g. weather, soil type). Disease pressure was enhanced by distributing F. graminearum colonized corn (0.5 gm/0.09 m 2 ) into plots prior to flowering. A misting system was also utilized to provide conditions favorable for disease development at the Macon Ridge Research Station. Seven treatments were evaluated in one test at the Macon Ridge location in : 1. Nontreated, 2. Folicur 4.0 fl. oz/a, 3. Proline 5.0 fl oz/a, 4. Prosaro (1:1 ratio of prothioconazole: 6.5 fl oz/a, fl oz/a, fl oz/a, and fl oz/a. Treatments were applied at early anthesis using a CO 2 charged hand-held boom delivering 17 gallons/a. Three treatments 1. Nontreated, 2. Proline 5.0 fl oz/a, and 3. Prosaro 5.0 fl oz/a were applied to 6 varieties (Terral LA841, D , Jamestown, GA E8, VA01W-433, and Pioneer 26R61. Treatments were applied at early anthesis using a CO 2 charged hand-held boom delivering 15 gallons/a. Scab epidemics did not develop at the Macon Ridge location; however, epidemics did develop at the Rice Research Station location. Yields, moisture, and test weights were reported from both locations. Data was analyzed using appropriate statistical procedures. Macon Ridge Research Station: No diseases were evident at the Macon Ridge location. Head samples were taken prior to harvest for DON analysis. Yields did not differ among treatments. Rice Research Station: Fungicides and varieties impact scab epidemics. When averaged across fungicide treatments, incidence ranged from 8.3% (VA01W-433) to 18.7% (Jamestown and GA E8). Severity ranged from 12.0% (D ) to 20.4% (Pioneer 26R61). The incidence, severity, and severity index in wheat treated with fungicides was less than that observed in the non-treated. When compared to the non-treated, yields were improved in Pioneer 26R61 treated with Proline. (Form FPR07) 2

269 PI: Padgett, Boyd USDA-ARS Agreement #: List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Efficacious fungicides are needed to manage scab. This collaborative research is attempting to identify fungicides effective for managing scab in wheat. Coordinating uniform tests throughout land grant universities in the U.S. will help expedite the identification of effective fungicides and application techniques for managing this disease. It was evident that fungicides can be used to reduce scab incidence and severity. It was also apparent that epidemics progressed at different rates within varieties. Therefore, this information should be utilized to develop an integrated strategy for managing scab. Impact: The discovery of effective fungicides and incorporation into wheat production systems will reduce the threat from scab and increase the quality of the U.S. crop. New fungicides efficacious against scab are being identified. In addition, these fungicides can be combined with genetic resistance (when available) to enhance scab management. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: This data will be added to an existing data to support decisions concerning the use of fungicides in wheat for managing scab. In addition, the results from these studies are necessary for obtaining an EPA approved commercial fungicide label (section 3). Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None (Form FPR07) 3

270 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Pierce Paul Institution: Ohio State University Address: Department of Plant Pathology 1680 Madison Ave. Wooster, OH Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 83,158 Splash Dispersal, Innoculum Level and Fungicide Effects on Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * CBCC EEDF ARS Adjusted Award Project Title Amount Integration and Economic Analysis of Control Strategies for FHB and DON in wheat. $39,256 Factors of Influencing Infection, DON Content, and FHB/DON Relationship in Wheat. $ 43,902 Total Award Amount $ 83,158 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

271 PI: Paul, Pierce USDA-ARS Agreement #: Project 1: Integration and Economic Analysis of Control Strategies for FHB and DON in wheat. 1. What major problem or issue is being resolved and how are you resolving it? Efforts to minimize the impact of Fusarium head blight (FHB) and its associated toxin (DON) have been based on the use of management strategies such as host resistance, crop rotation, tillage, and fungicide application. However, none of these methods used alone have been fully effective against FHB and DON. The effects of fungicide application, genetic resistance, and residue management (through crop rotation or tillage) are highly variable and strongly influenced by the environment. Moderately resistant varieties may still become infected, with DON contamination exceeding critical threshold levels. Fungicide efficacy varies from one trial to another, with mean percent control of approximately 50% for FHB and 40% for DON, and yield and quality gains are not always enough to offset the cost of fungicide application. Coordinated research is needed to evaluate the efficacy and economics of using multiple approaches to management FHB and DON. Following a standard protocol, field experiments were conducted across major US wheat-growing regions to evaluate the integrated effects of genetic resistance and fungicide application on FHB and DON. In Ohio, pairs of plots of six soft red winter wheat varieties were planted in a split-plot treatment layout in a randomized complete block design, and one plot of each variety was treated with a fungicide (3 fl oz/a Proline + 3 fl oz/a Folicur) at anthesis (Feekes growth stage ). Incidence and severity ( index ) of FHB were assessed in each plot at early dough (Feekes GS 11.2) and a sample of harvested grain was tested for DON. A second experiment was conducted to evaluate the effects of seeding rate (18, 23, and 28 seeds per foot of row), row spacing (7- and 15-inch rows), and fungicide application on FHB and DON. The experimental design was a split-split plot with 3 replicate blocks. Row spacing served as the whole-plot factor, seeding rate as the sub-plot factor, and fungicide application (with and without a single application of Prosaro at 6.5 fl. oz/a % Induce) as the sub-sub plot factor. Data from all integrated management trials were sent to Ohio for analysis and synthesis. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Due of unfavorable weather conditions during flowering and early grain fill, disease levels were low in Ohio. During the first two weeks of May, between 0 and 0.8 inches of rainfall were recorded. At the OARDC where the trials were conducted, the wheat crop flowered between May 22 and 29, a period of warm, dry weather conditions. FHB intensity and grain DON content were very low in all plots, with incidence less that 1% and mean DON contamination less than 0.2 ppm. A total of 14 additional FHB integrated management trials were conducted across the US in Based on the results from those trials in which some level of FHB developed (more that 5% index in the untreated check), both the main and interaction effects of variety and fungicides were statistically significant (P < 0.005). In general, moderately resistant variety + fungicide (Form FPR07) 2

272 PI: Paul, Pierce USDA-ARS Agreement #: treatment combination resulted in the highest percent control of FHB and DON. For those trials with cropping sequence as a treatment factor, none-host crop + moderately resistant variety + fungicide generally resulted in the highest percent control Linear mixed model analysis of the results from experiment 2 (conducted in Ohio) revealed that the main effects of row spacing (7 and 15 inches between rows) and planting density (seeds per foot of row) on yield, test weight, and spike counts (number of spikes per foot of row) were not statistically significant. However, the effect of row spacing on spring stand count (tillers per foot of row) was significant (P < 0.05), suggesting that row spacing and planting density may be manipulated to alter the architecture of the crop canopy under conditions in Ohio (in some years) without negatively impacting grain yield. Impact: The 2007 growing season was the first in which coordinated FHB integrated management trials were conducted across the US. Findings from trials with disease clearly indicated that the integration of multiple (and potentially complementary) approaches such as crop rotation, resistance, and fungicide application would provide the most effect control of FHB and DON. Results from the row-spacing x planting density study suggest that the architecture of the wheat canopy may be altered without negatively impacting yield and quality. Modifying these cultural practices may improve fungicide coverage, and as such, the overall FHB and DON control efficacy with fungicide. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The ultimate goal of the integrated management program is to develop and provide growers with a series of best management strategies for FHB and DON to help minimize yield and quality losses. A quantitative synthesis of the results from these trials will be made available via the internet and extension publications. Trials conducted at multiple locations will allow for the evaluation of similar integrated strategies under a range of environmental conditions, potentially reducing the time needed for the establishment of the best management strategies. Findings from these trials may also lead to the development of location- or region-specific management recommendations for FHB and DON. (Form FPR07) 3

273 PI: Paul, Pierce USDA-ARS Agreement #: Project 2: Factors of Influencing Infection, DON Content, and FHB/DON Relationship in Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Critical for the epidemiology group s goals of adding a DON module to the FHB risk assessment model and developing mechanistic FHB and DON models are an understanding of the relationship between visual symptoms of FHB and DON accumulation and a quantitative measure of differences among varieties in terms of DON accumulation. Through the cooperative epidemiology effort, valuable information has been gathered about temperature and moisture effects on perithecia development and inoculum abundance, and the influence of rainfall intensity on splash dispersal of inoculum within wheat canopies. However, less is known about specific conditions required for spore survival, deposition, colonization, and DON accumulation. It is widely known that moderate to high temperatures, high relative humidity, and rainfall during anthesis favor FHB development (manifestation of visual symptoms) and DON contamination. However, conditions favorable for infections associated with moderate to low symptom manifestation and disproportionately high DON accumulation is less clearly understood. DON may exceed threshold levels even when disease symptoms are low or absent. In addition, the relative response of commercial varieties to DON accumulation has not been characterized under field conditions. Two experiments were designed to investigate factors influencing FHB infection and DON accumulation and the FHB/DON relationship. In the first, a split-split plot design was used with planting date (Hessian Fly-safe date and two weeks later) as the whole plot, variety (Truman, resistant; Cooper, susceptible; and Hopewell, moderately susceptible) as the sub plot, and inoculation timing (early and late anthesis) as the sub-sub plot. In the second, three plots each of six soft red winter wheat varieties were planted in each of three blocks, in a split plot design. One plot of each variety was left untreated, one treated with Folicur 3.6F (4 fl. oz./a % Induce), and the third treated with Prosaro (6.5 fl. oz/a % Induce). Incidence and severity ( index ) of FHB were assessed within each plot at early dough (Feekes GS 11.2) and a sample of harvested grain was tested for DON. In addition, spikes in each of 11 disease index categories (0, 5, 10, 15, 20, 25,, and 50%) were tagged in each plot and harvested for DON and RT-PCR analysis. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Due of unfavorable weather conditions during flowering and early grain fill, disease levels were low in non-inoculated plots. However, FHB developed well in inoculated plots. For experiment one, the main effects of inoculation timing, planting date, and variety and all interactions involving these factors on FHB index were statistically significant (P < 0.05). However, for incidence and DON, the effect of planting data was not significant (at the 5% level of probability). Significant interaction between variety and inoculation timing suggest that, depending on when infection occurs relative to anthesis, varieties with different levels of resistance to FHB may have similar levels of disease and toxin accumulation. For instance, the difference in DON accumulation between Cooper, the susceptible variety, and Truman, the moderately resistance variety, was not statistically significant when Truman was inoculated at (Form FPR07) 4

274 PI: Paul, Pierce USDA-ARS Agreement #: late anthesis and Cooper was inoculated at early anthesis, however, when Truman was inoculated at early anthesis and Cooper inoculated at late anthesis, the difference in DON contamination between the two varieties was statistically significant (P < 0.005). For the second experiment, results from an analysis of covariance showed that DON content (ppm) increased with increasing FHB index in all three varieties. However, the rate of change in DON with change in severity (the regression slope) was generally greater for the susceptible varieties than the resistant variety. The magnitude of the difference in DON content among the varieties, at a given level of severity, was higher at high severity (30% index) than at low severity (5% index). Contrastingly, estimated slopes for relationships between fungal biomass (log-transformed ng/mg) and DON (ppm) were similar for the three varieties, suggesting similarity among the varieties in the rate of increase in DON content with increase in fungal colonization. However, the heights of the regression lines for the fungal biomass/don relationships differed among the varieties, indicating that for a similar level of fungal colonization, DON accumulation differed among the varieties. Although based on visual symptoms Cooper is more susceptible to FHB than Truman, Cooper had DON content that was lower than, or comparable to, that of Truman for a given fungal biomass, under the conditions of this study. Impact: Data from these experiments are being used to validate and refine existing risk assessment models for FHB and to develop mechanistic FHB and DON risk assessment models. The web-based FHB models are currently being used in 24 US states as an early warning system to prepare producers, grain buyers, and the milling industry for possible epidemics of FHB and to help producers make fungicide application decisions. Results from experiment two will serve as the basis for further investigations of the associations among FHB, DON and variety. This may provide insight as to the mechanisms involved in resistance to FHB and DON in soft red winter wheat varieties. The findings from experiment two suggest that resistance to FHB does not necessarily parallel resistance to DON accumulation. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: With new and fairly effective fungicides recently registered for FHB and DON management, the web-based risk assessment models are now an important part of integrated management programs for this disease and toxin. More growers and crop consultants will routinely refer to these models to make management and marketing decisions. Improving the prediction accuracy of the risk models, adding a DON prediction module to the web-based risk tool, and characterizing varieties in terms of their differential DON accumulation will all contribute to improving the efficacy FHB and DON management. (Form FPR07) 5

275 PI: Paul, Pierce USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-Reviewed Articles Paul, P. A., Lipps, P. E., De Wolf, E., Shaner, G., Buechley, G., Adhikari, T., Ali, S., Stein, J., Osborne, L., and Madden, L. V A distributed lag analysis of the relationship between Gibberella zeae inoculum density on wheat spikes and weather variables. Phytopathology 97: Paul, P. A., Lipps, P. E., Hershman, D. E., McMullen, M. P., Draper, M. A., and Madden, L. V A quantitative review of tebuconazole effect on Fusarium head blight and deoxynivalenol content in wheat. Phytopathology 97: Abstracts Paul, P. A., Lipps, P. E., Hershman, D. E., McMullen, M. P., Draper, M. A., and Madden, L. V Relative efficacy of triazole-based fungicides for Fusarium head blight and deoxynivalenol control in wheat. Phytopathology 97:S90. Nita, M., DeWolf, E., Madden, L., Paul, P., Shaner, G., Adhikari, T., Ali, S., Stein, J. and Osborne, L Integrated management of Fusarium head blight (FHB) and deoxynivalenol contamination. Phytopathology 97:S85. Proceedings and Presentations Paul, P., Lipps, P., Hershman, D., McMullen, M., Draper, M., and Madden, L A Quantitative synthesis of the relative efficacy of triazole-based fungicides for FHB and DON control in wheat. Pages in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Salgado, J. D., Broders, G., Madden, L., and Paul, P Characterization of DON accumulation in SRWW cultivars with different levels of type II resistance to FHB. Page 137 in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Wallhead, M., Madden, L., and Paul, P Differential sensitivity to triazole-based Fungicides among Isolates of Fusarium graminearum. Page 141 in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Paul, P., Madden, L., McMullen, M., Hershman, D., Sweets, L., Wegulo, S., Bockus, W., Halley, S., and Ruden, K An integrated approach to managing FHB and DON in wheat: uniform trials Pages in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. (Form FPR07) 6

276 PI: Paul, Pierce USDA-ARS Agreement #: Paul, P. A., Madden, L. V., Wegulo, S., Adhikari, T., Ali, S., and De Wolf, E Influence of SRWW, HRSW, and HRWW varieties on the relationship between FHB and DON. Page 128 in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Paul, P., Madden, L., McMullen, M., Hershman, D., Brown-Rytlewski, D., Sweets, L., Adee, E., Padgett, B., and Ruden, K Fungicide effects on FHB and DON in wheat across multiple locations and wheat classes: uniform fungicide trials Pages in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Nita, M., DeWolf, E., Madden, L., Paul, P., Shaner, G., Adhikari, T., Ali, S., Stein, J., Osborne, L., and Wegulo, S Mechanistic simulation models for Fusarium head blight and deoxynivalenol. Page 108 in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Sneller, C., Paul, P., Herald, L., Sugerman, B., Johnston, A Report on the northern uniform winter wheat scab nurseries (NUWWSN and PNUWWSN). Pages in: Proc Natl. Fusarium Head Blight Forum, Kansas City, MO. Extension Newsletters (Electronic) Pierce Paul and Dennis Mills - Problems with Planting Wheat After Wheat or Wheat After Corn. Crop Observation and Recommendation Network (C.O.R.N.) Pierce Paul and Dennis Mill - Risk of Head Scab Low. Crop Observation and Recommendation Network (C.O.R.N.) Pierce Paul and Dennis Mill - Conditions not Favorable for Foliar Disease Development. Crop Observation and Recommendation Network (C.O.R.N.) Pierce Paul and Dennis Mill - Managing Wheat Diseases with Fungicides. Conditions not Favorable for Foliar Disease Development. Crop Observation and Recommendation Network (C.O.R.N.) Pierce Paul and Dennis Mill - Wheat Scab Risk Prediction Crop Observation and Recommendation Network (C.O.R.N.) (Form FPR07) 7

277 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: James Pestka Institution: Michigan State University Address: Department of Food Science & Human Nutrition 234A GM Trout FSHN Bldg. East Lansing, MI Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Human Susceptibility to Trichothecene Mycotoxins. Title: FY07 ARS Award Amount: $ 93,991 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount FSTU-R Human Susceptibility to Trichothecene Mycotoxins. $93,991 Total Award Amount $ 93,991 Principal Investigator 7/14/2008 Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

278 PI: Pestka, James USDA-ARS Agreement #: Project 1: Human Susceptibility to Trichothecene Mycotoxins. 1. What major problem or issue is being resolved and how are you resolving it? During head blight of wheat and barley, deoxynivalenol (DON or vomitoxin ) and other trichothecene mycotoxins are elaborated that might adversely affect human health. Two aspects related to trichothecene exposure and human health were addressed. First, since DON can potentially cause adverse health effects in individuals who consume the infected grain, this mycotoxin DON is regulated in the U.S. at 1 ppm in finished food. Recently, the European Economic Union have established lower limits based on a a limited number of rodent studies as well as the concern that children have greater sensitivity to DON. A second concern is that agricultural workers are exposed to airborne DON during harvest, threshing and milling of infected wheat and barley, however, we do not understand the potential for adverse effects from inhalation of this toxin. Overall, serious questions remain relative to the true risks presented to consumers and grain handlers posed by DON and other trichothecenes elicited during outbreaks head blight. Our research is addressing this gap by providing data on effects of DON on toxicity biomarkers using the young (weanling) mouse model and a mouse intranasal instillation model. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment #1: We determined that the initial plasma and tissue DON concentrations in young mice were approximately twice that of adult mice given identical doses of the toxin but that clearance was rapid in both groups of mice. DON induction of TNF-α, IL-1β and IL-6 in young animals were two to three times greater in spleens, a primary source of systemic proinflammatory cytokines, whereas differences in proinflammatory cytokine expression were less robust or not apparent in the liver or lung between the two age groups. Impact: This is the first report comparing the effects of age on trichothecene uptake and molecular endpoints of toxicity. Mice have been previously used to model potential adverse effects of DON to humans and large safety factor (100X) for species differences (10X) and human variability (10X) have been employed in Tolerable Daily Intake (TDI) estimates. The differences seen in young mice compared to adult mice relative to uptake and toxicity are likely accounted for in this large safety factor. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The effects of age on DON uptake and toxicity biomarkers were quite modest. They do not support the need to lower the level of concern for DON in U.S.food because of special risks to children. These findings are being published in Food and Chemical Toxicology. (Form FPR07) 2

279 PI: Pestka, James USDA-ARS Agreement #: Accomplishment #2: We have determined that DON distributes rapidly throughout mouse organs and reaches peak plasma concentration within 30 minutes of exposure regardless of whether mice are exposed to the toxin via gut or respiratory route. However, plasma and tissue DON concentrations in mice exposed by intranasal route were 1.5 to 3 times that of mice orally exposed to DON. Relative inductions of cytokine biomarkers were 2 to 10 times that observed in the tissues of orally exposed mice. Taken together, these data suggest that DON is more toxic when inhaled than when ingested, and further suggest the potential for adverse human health effects from inhalation of DON-contaminated grain dust. Impact: Quantification of kinetic differences between nasal and oral routes provides a basis for characterization of potential risk(s) associated with DON inhalation. These observations show for the first time a potential health effect with respiratory exposure to DON suggesting the need for adequate risk characterization and risk management for grain handlers. Overall this brings us closer to understanding the effects of DON inhalation in a quantifiable manner As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? There is a possible hazard to agricultural workers handling Fusarium-contaminated grain containing very high DON levelshas been identified. This is an important consideration in for establishing safety measures for workers occupationally exposed to DON during grain handling. These findings are being published in Toxicology Letters. Accomplishment #3: The eukaryotic ribosome has long been known to be a target of DON and other trichothecenes. We employed cell-free and the clonal macrophage models to test the hypothesis that DON and T-2 exposure mediates cleavage of rrna. The results suggest that while these trichothecenes did not possess the inherent enzymatic activity in a cell-free model, DON and T-2 evoked 28S rrna cleavage in the macrophage, possibly by facilitating the action of constitutive and induced RNases. Impact: This is the first report of specific ribosomal rrna cleavage by trichothecenes. Pending further work, this lesion has potential use as a biomarker of for DON exposure and toxicity on animals and humans. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before: A cellspecific type of damage due to DON has been identified in cells of the immune system- a central tissue target for this toxin.. These findings are being published in Toxicology Sciences. This information has the potential to be used safety studies employing animals as well as measuring in human exposure. (Form FPR07) 3

280 PI: Pestka, James USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. (1) Pestka JJ, Islam Z, Amuzie CJ. Immunochemical assessment of deoxynivalenol tissue distribution following oral exposure in the mouse. Toxicol Lett 2008 May 5; 178(2):83-7, PM: (2) Pestka JJ, Amuzie CJ. Tissue distribution and proinflammatory cytokine gene expression following acute oral exposure to deoxynivalenol: Comparison of weanling and adult mice. Food and Chemical Toxicol (in press) 2008 (3) Amuzie CJ, Harkema JR, Pestka JJ. Tissue distribution and proinflammatory cytokine induction by the trichothecene deoxynivalenol in the mouse: Comparison of nasal vs. oral exposure. Toxicology 2008 Jun 3; 248(1):39-44, PM: (4) Li M, Pestka JJ. Comparative induction of 28S ribosomal RNA cleavage by ricin and the trichothecenes deoxynivalenol and T-2 toxin in the macrophage. Toxicol Sci 2008 Jun 4; PM: (5) Pestka JJ. Mechanisms of deoxynivalenol-induced gene expression and apoptosis. Food Addit Contam (in press) 2008; (6) Pestka JJ. Deoxynivalenol: Toxicity, mechanisms and animal health risks. Animal Feed Science and Technology 2007; 137: (7) Gray JS, Pestka JJ. Transcriptional regulation of deoxynivalenol-induced IL-8 expression in human monocytes. Toxicol Sci 2007 Oct; 99(2):502-11, PM: (8) Gray JS, Bae H-K, Li JCB, Lau AS, Pestka JJ. Double-stranded RNA-activated proteiin kinase (PKR) mediates induction of IL-8 expression by deoxynivalenol, Shiga toxin 1 and ricin in monocytes. Toxicol Sci (in press) 2008; (Form FPR07) 4

281 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Joel Ransom Institution: North Dakota State University Address: Department of Plant Sciences 266H Loftsgard Hall Fargo, ND Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 16,462 Integrated Management of FHB in Spring and Winter Wheat in North Dakota. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount CBCC Integrated Management of FHB in Wheat and Barley. $16,462 Total Award Amount $ 16,462 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

282 PI: Ransom, Joel USDA-ARS Agreement #: Project 1: Integrated Management of FHB in Wheat and Barley. 1. What major problem or issue is being resolved and how are you resolving it? Currently no single factor can control FHB in wheat and barley, so an integrated disease management approach is needed. How the crop is planted (plant density and row spacing) may play a role in the development of FHB and therefore could be a component of an integrated control program. Experiments were conducted during the 2007 growing season in Fargo, Prosper and Minot North Dakota to evaluate the influence of row spacing and plant density, when combined with fungicides and resistant cultivars on FHB severity in wheat and barley and on yield and crop quality. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Methodology for establishing small plots where differing row spacing and plant populations could be tested were developed and implemented in The 2007 growing season was dry and hot during flowering, so FHB did not develop in any of the experiments. However, we were able to quantify the influence of seeding rates and row spacings on yield in the absence of disease. Reduced seeding rates and increased row spacings were found to slightly reduce yields this season. These data give us a baseline by which we can compare yield responses to management practices in a relatively dry year to those when FHB is prevalent. Impact: The impact of this activity has been in the development of data that quantify the effects of the various practices evaluated on yield so that as additional data are developed over additional environments, integrated crop and disease management recommendations can be extrapolated with more confidence. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The scientific community now has information on the possible effects of management practices on yield. Particularly, that row spacing and plant populations can be altered significantly with only minimal yield reductions, thereby offering an additional tool that can be used to reduce disease build up. (Form FPR07) 2

283 PI: Ransom, Joel USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Ransom, J. Response of Winter Wheat Genotypes to Fungicide Applied at Flowering in Eastern North Dakota. Poster presentation at the North American Wheat Workers meeting, Saskatoon, Canada, March Ransom, J.K., M.P. McMullen and S. Meyer Effect of fungicides on FHB control and yield of winter wheat cultivars in North Dakota. Proceedings of the 2007 National Head Blight Forum, p Poster and published abstract. Ransom, J.K., and M. McMullen Yield and disease control on hard winter wheat cultivars with foliar fungicides. Agronomy Journal 100: (in press) (Form FPR07) 3

284 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Ivan Rayment Institution: University of Wisconsin Address: Deptartment of Biochemistry 433 Babcock Drive Madison, WI Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 52,683 Structural and Functional Studies of Trichothecene Biosynthetic Enzymes. USWBSI Individual Project(s) USWBSI Research Area * PGG ARS Adjusted Award Project Title Amount Development of Improved Enzymes for the Inactivation of $52,683 Trichothecene Toxins. Total Award Amount $ 52,683 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

285 PI: Rayment, Ivan USDA-ARS Agreement #: Project 1: Development of Improved Enzymes for the Inactivation of Trichothecene Toxins. 1. What major problem or issue is being resolved and how are you resolving it? The major goal for the past fiscal year was to develop an improved enzyme for inactivating fungal mycotoxins. It was planned to utilize the three-dimensional structure of trichothecene 3-O-acetylase from F. sporotrichioides and F. graminearum (Tri101) as a framework to develop a modified enzyme with improved efficacy towards the inactivation of DON and nivalenol. Progress towards this goal was assisted by a kinetic analysis of these isozymes that clearly indicated that an improved enzyme could be made. The work utilizes a combination of biomolecular structural engineering coupled with kinetic and thermodynamic measurements. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The most important accomplishment was the determination of the structure of trichothecene 3-O-acetylase from F. sporotrichioides and F. graminearum (Tri101) coupled with an analysis of the kinetic properties of these enzymes. This has allowed improved enzymes to be designed, developed, and tested that show greater thermal stability than the wild type enzyme and a better kinetic profile for the toxins found in the USA. Impact: The improved enzymes have the potential to create a transgenic barley or wheat that shows greater resistance to FHB. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: As a consequence of the studies performed in this proposal it is possible to propose why the earlier transgenic wheat and barley that utilized Tri101 were not effective. These studies provide a better enzyme for inactivating trichothecene mycotoxins which can be tested in future studies. (Form FPR07) 2

286 PI: Rayment, Ivan USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. J Biol Chem Jan 18;283(3): Epub 2007 Oct 8. Structural and functional characterization of the TRI101 trichothecene 3-O-acetyltransferase from Fusarium sporotrichioides and Fusarium graminearum: kinetic insights to combating Fusarium head blight. Garvey GS, McCormick SP, Rayment I. (Form FPR07) 3

287 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: David Schmale Institution: Virginia Polytechnic Institute and State University Address: Dept. of Plant Pathology, Physiology, Weed Science 410 Price Hall Blacksburg, VA Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 24,390 Contribution of Local Inoculum Sources to Regional Atmospheric Populations of G. zeae. USWBSI Individual Project(s) USWBSI Research Area * EEDF ARS Adjusted Award Project Title Amount Contribution of Local Inoculum Sources to Regional Atmospheric $24,390 Populations of G. zeae. Total Award Amount $ 24,390 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

288 PI: Schmale, David USDA-ARS Agreement #: Project 1: Contribution of Local Inoculum Sources to Regional Atmospheric Populations of G. zeae. 1. What major problem or issue is being resolved and how are you resolving it? Epidemics of FHB may be initiated by regional atmospheric sources of inoculum of G. zeae, but little is known about the contribution of local inoculum sources to regional atmospheric populations of the pathogen and how far inoculum may travel from its source. The ability to predict the regional transport of G. zeae from local inoculum sources may help refine risk models for FHB. The ultimate goals of our research efforts are to determine where inoculum for FHB comes from and how far it travels. We used unmanned aerial vehicles (UAVs), DNA fingerprinting, and the atmospheric transport model HYSPLIT to predict and track the movement of G. zeae in the atmosphere. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Little is known about the contribution of local inoculum sources to regional atmospheric populations of the G. zeae and how far inoculum may travel from its source. We inoculated a portion of a tilled field with corn kernels colonized with a single clonal isolate of G. zeae, and we conducted 35 autonomous UAV sampling flights for the G. zeae 100 m above our inoculum source. Isolates of G. zeae were cultured from the sampling plates, and a DNA fingerprinting technique known as Amplified Fragment Length Polymorphisms (AFLPs) was used to track the released isolate in our atmospheric collections. The atmospheric transport model HYSPLIT was used to forecast the long-distance transport of G. zeae from our inoculum source. Thirty three percent of our isolates were collected on two different flights on May 1, Runs with HYSPLIT for these collection times suggested that our inoculum source was transported at least a kilometer away from the ground surface within an hour. The high number of colonies recovered during this time period, coupled with data from AFLP analyses, is consistent with HYSPLIT model predictions of spore transport. Impact: This is the first study to track the movement of G. zeae in the atmosphere from a clonal inoculum source, and the first body of work to predict the long-distance transport of G. zeae over broad geographical regions. This project has developed new and improved tools (e.g., autonomous UAVs) necessary for studying regional atmospheric populations of G. zeae, and has assisted in predicting the distribution and spread of G. zeae in the atmosphere. Our work continues to aid in developing and/or excluding strategies for managing FHB, and should help researchers refine forecasting/risk assessment models for regional FHB epidemics. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Knowledge of the long-distance transport of G. zeae from local inoculum sources is an important step toward developing an early warning system for FHB and practical management strategies for FHB epidemics. Since spores of G. zeae may move over kilometer distances in the atmosphere, management strategies for FHB should be considered on a regional (i.e. county or state) basis. Forecasting and risk assessment models for FHB epidemics could benefit from the inclusion of information on atmospheric inoculum dynamics. (Form FPR07) 2

289 PI: Schmale, David USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Presentations Schmale, D.G Use of autonomous unmanned aircraft to validate and improve long distance pathogen transport models. Phytopathology 97: S147. Bergstrom, G.C., and Schmale, D.G The aerobiology of Gibberella zeae: whence come the spores for Fusarium head blight? Pages: in Proc National Fusarium Head Blight Forum, Kansas City, MO. Schmale, D.G Bridging engineering and agriculture at Virginia Tech s Kentland Farm. Field Day in association with the2 007 Potomac Division Meeting of the American Phytopathological Society. Peer-reviewed articles Schmale, D.G., Dingus, B.R., and Reinholtz, C Development and application of an autonomous unmanned aerial vehicle for precise aerobiological sampling above agricultural fields. Journal of Field Robotics 25: Wang J., Patel V., Woolsey C.A., Hovakimyan N., and Schmale D.G L1 adaptive control of a UAV for aerobiological sampling. American Control Conference, Non peer-reviewed articles Schmale, D. G., Dingus, B.R., Wood-Jones, A. K., Khatibi, P, and Reinholtz, C Diversity of the fungal genus Fusarium one hundred meters above the surface of the earth. Page 46 in: 2007 Dean s Forum on the Environment, Virginia Tech, Blacksburg, VA. Keller, M., Duttweiler, K., Schmale, D.G., and Bergstrom, G.C Contribution of withinfield inoculum sources to Fusarium Head Blight in wheat. Page 98 in Proc National Fusarium Head Blight Forum, Kansas City, MO. Schmale, D.G., Dingus, B.R., Keller, M., and Wood-Jones, A.K Contribution of local inoculum sources to regional atmospheric populations of Gibberella zeae. Page 139 in Proc National Fusarium Head Blight Forum, Kansas City, MO. (Form FPR07) 3

290 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Paul Schwarz Institution: North Dakota State University Address: Department of Plant Sciences P.O. Box 5051 Fargo, ND Phone: Fax: or Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Malting Barley Deoxynivalenol Diagnostic Services. Title: FY07 ARS Award Amount: $ 90,143 USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount FSTU-S Malting Barley Deoxynivalenol Diagnostic Services. $90,143 Total Award Amount $ 90,143 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

291 PI: Schwarz, Paul USDA-ARS Agreement #: Project 1: Malting Barley Deoxynivalenol Diagnostic Services. 1. What major problem or issue is being resolved and how are you resolving it? Mycotoxin analyses are essential for most researchers working on FHB of cereals. However, in barley DON is a major economic factor, and new varieties must display increased resistance to DON accumulation as well as to FHB. Screening barley lines for DON is requisite for any breeding program intending to develop varieties for the upper Midwestern USA. DON analytical services are primarily provided to three barley varietial developmental programs. These breeding programs stated a need for the analysis of approximately 12,000 samples in FY07. Supporting research and extension work requires an additional 3,000 to 4,000 samples. In total, fifteen collaborating scientists were served. The major issue is to provide DON analytical services in a cost effective, timely and accurate manner. Funds provided by the USWBSI have allowed us to hire additional personnel and to subsidize the cost of analysis. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Approximately 15,000 samples (excluding standards) were analyzed from May 2007 to April These analyses were primarily from barley varietial development programs (n=11,560). Additional analysis were conducted for personnel involved in extension/crop production work (n=1721) and for barley FHB research projects. Samples analyzed as part of the 2007 regional barley crop quality survey indicated that average levels of DON were among the lowest observed since testing began in Our associated research work on DON analysis has demonstrated that some barley samples may contain a significant portion of bound DON. The 2007 samples were analyzed between August, 2007 and July, Impact: This project provides essential support to all barley breeding programs working on the development of FHB-resistant varieties for the Midwestern USA. Our findings on the occurrence of bound DON in some barley samples should be of interest to both feed/food processors and to researchers. Binding or conjugation of DON may be involved in FHB resistance, and studies involving enzyme pretreatments suggest it might be released during food processing or in digestion. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Timely and accurate analyses allow the breeders and support scientists to make advances in achieving the goal of developing new varieties that display reduced levels of DON accumulation. Continued monitoring of DON levels in regional grain (crop survey) is important in maintaining food and feed safety, and also helps to maintain an epidemiological record of FHB on barley in the upper Midwest. This data is important to producers and users of regional grain, pathologists, agricultural economists, and to regulatory agencies. Publication of three peer-reviewed manuscripts on bound DON in barley has made the (Form FPR07) 2

292 PI: Schwarz, Paul USDA-ARS Agreement #: results of our research in this area widely available to research scientists and food processors. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Publications: Zhou, B., He, G.Q., and Schwarz, P.B. Occurrence of bound deoxynivalenol in Fusarium Head Blight infected barley (Hordeum vulgare L.) and malt as determined by solvolysis with trifluoroacetic Acid. J. Food Protection 71(6): , Zhou, B., Schwarz, P.B., He, G.Q., Li, Y., Gillespie, J., and Horsley, R. Effect of enzyme pretreatments on the determination of deoxynivalenol in barley. J. Am. Soc. Brew. Chem. 66(2): , Zhou, B., Li, Y., Gillespie, J., He, G.Q., Horsley, R., and Schwarz, P.B. Doehlert matrix design for optimization of the determination of bound deoxynivalenol in barley grain with TFA. J. Ag. Food Chem. 55, , Presentations: Boddeda, A., Schwarz, P., and Wolf-Hall1, C. Quantification of The Tri5 Gene, Expression and Deoxynivalenol Production During the Malting of Barley. Poster National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. Schwarz, P., Dong, Y., and Dill-Macky, R. Deoxynivalenol Measurement: Sources of error and Sampling Recommendations. Oral Presentation National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. Tobias, D.J., Vashisht, A., Boddeda1, A., Wolf-Hall, C., and Schwarz, P. Development of a Multiplex Real-Time Pcr Assay for Rapid Detection and Quantification of Fusarium Spp. in Barley National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. Tobias, D.J., Wolf-Hall, C., and Schwarz, P. Ozone as an Antimycotic Agent in Malting Barley National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. Zhou, B., Li, Y., Gillespie, J., Horsley, R., and Schwarz, P. Doehlert Matrix Design for Optimization of the Determination of Bound Deoxynivalenol in Barley Grain with TFA National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. Zhou, B., Gillespie, J., Horsley, R., and Schwarz, P. Effect of Enzyme Pretreatments on the determination of Deoxynivalenol in Barley National Fusarium Head Blight Forum. December 2-4, Kansas City, MO. (Form FPR07) 3

293 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Steve Scofield Institution: USDA-ARS Address: Crop Prod. & Pest Control Research 915 West State Street West Lafayette, IN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: NA USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 55,000 Rapid Function Identification of Genes Contributing to FHB Resistance and Susceptibility. USWBSI Individual Project(s) USWBSI Research Area * HGG ARS Adjusted Award Project Title Amount Rapid Function Identification of Genes Contributing to FHB $55,000 Resistance and Susceptibility. Total Award Amount $ 55,000 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

294 PI: Scofield, Steve USDA-ARS Agreement #: NA Project 1: Rapid Function Identification of Genes Contributing to FHB Resistance and Susceptibility. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) is one of the major diseases that threaten US wheat and barley producers. Unfortunately, all known resistance to FHB is complex, based on multiple quantitative trait loci (QTL) that make partial contributions to resistance. Combining these QTL loci into high yielding wheat lines and eliminating the yield reducing traits linked to some of the QTLs has proven to be a slow process. Development of resistant varieties would be accelerated by knowledge of the specific genes that make essential contributions to FHB resistance. We have developed a virus-induced gene silencing (VIGS) system based on Barley Stripe Mosaic virus (BSMV) that can switch-off, or silence, genes in hexaploid wheat so that the gene s function can be inferred by observing the effects of its silencing. Because VIGS is homology-dependent, it can silence related gene copies that encode mrnas with >85% homology, making it ideal for creating knockouts in hexaploid wheat where at least three copies of each gene may be expressed from the A, B and D genomes. We have demonstrated the utility of BSMV-VIGS for functionally identifying genes involve in several disease resistance systems in hexaploid wheat. In these studies a candidate gene is silenced in a plant that is normally resistant to a particular pathogen. After silencing is established, the plant is challenge with this pathogen. If the plant becomes susceptible, we have strong evidence that the candidate gene has an essential function in this resistance pathway. We have developed protocols for silencing genes in spikes of wheat plants and are now screening a list of candidate genes testing if they contribute to FHB resistance. It is very important to note, that at the start of this work, no wheat or barley gene was know that made a clear functional contribution to FHB resistance. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Our most important accomplishment in the work funded by the FY06 award was the discovery that wheat chitinase genes make an essential contribution to FHB resistance. This result is very important for at least two reasons. 1. It validates the ability of the VIGS assay to rapidly identify genes contributing to FHB resistance and provides confidence that continuing this approach can identify the key genes in this resistance mechanism. 2. Knowing that induction of chitinase activity makes a crucial contribution to FHB resistance gives us important insight into how the wheat and almost certainly barley defend against Fusarium. Impact: The FHB research community now has a powerful tool that can be used to determine which genes have essential functions in FHB resistance. These genes can then be targeted to find the best alleles in breeding strategies or employed in transgenic solutions to improving FHB resistance. (Form FPR07) 2

295 PI: Scofield, Steve USDA-ARS Agreement #: NA As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The importance of this new system is evidenced by the fact that the leading labs in FHB research are adopting this technique for their own work. To this end my lab has hosted and trained scientists in the FHB-VIGS technique from Gary Muehlbauer s lab (University of Minnesota), Kim Hammond-Kosak s lab, Rothamsted, UK) and Therese Ouellet, (Agriculture and Food Canada, Ottawa, Canada) Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. 1. National Wheat Genomics Conference, held November 30 December 2, 2007, in Kansas City. 2. US Wheat and Barley Scab Initiative National Forum, held in Kansas City from December 2 5, The Fifth Canadian Workshop on Fusarium Head Blight, held in Winnepeg, Canada November 27-30, I have been invited to speak at the 3 rd International Fusarium Head Blight Symposium in Szeged, Hungary, Sept (Form FPR07) 3

296 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Jyoti Shah Institution: Kansas State University (FY07 Award Period) Address: C/O University of North Texas Department of Biological Sciences Denton, TX Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 88,722 Engineering Fusarium Head Blight Resistance and Plant Defense Signaling. USWBSI Individual Project(s) USWBSI Research Area * GET HGG ARS Adjusted Award Project Title Amount Engineering Scab Resistance in Wheat with Defense Signaling Genes. $46,185 Signaling Mechanism in Host Defense and Susceptibility to Fusarium graminearum. $ 42,537 Total Award Amount $ 88,722 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

297 PI: Shah, Jyoti USDA-ARS Agreement #: Project 1: Engineering Scab Resistance in Wheat with Defense Signaling Genes. 1. What major problem or issue is being resolved and how are you resolving it? Losses to Fusarium head blight (FHB; also known as Scab) disease of wheat and barley have averaged $ million per annum. Fusarium graminearum is the leading agent of FHB in the US. Monogenic resistance against FHB is not known and current control methods utilize a combination of planting partially resistant varieties with fungicide application and crop rotation. Genetic engineering provides an alternative approach for developing wheat and barley germplasms with heightened resistance to FHB. It provides the advantage that novel genes and chimeras that are not currently in the partially resistant germplasms, can be introduced into wheat and barley, thus adding to the repertoire of genes that can be utilized in breeding programs for enhancing FHB resistance. Previously, ectopic expression of the Arabidopsis thaliana NPR1 (AtNPR1) gene from the maize ubiquitin promoter was shown to enhance FHB resistance in the partially FHB-resistant cv. Bobwhite under greenhouse conditions and in two field trials conducted in Kansas. NPR1 controls the activation of salicylic acid-dependent defense responses in plants, which our studies have demonstrated, is important for resistance to F. graminearum in Arabidopsis thaliana. PAD4 is another gene from Arabidopsis thaliana that enhances resistance to F. graminearum. PAD4 modulates salicylic acid synthesis in pathogen inoculated plants and production of antimicrobial phytoalexins. In addition, it controls other poorly defined defense mechanisms. As part of this USDA-ARS USWBSI-sponsored project we have continued field trials of AtNPR1 expressing cv. Bobwhite and have introduced the AtNPR1-conferred Scab resistance into two durum cultivars. Furthermore, we have engineered AtPAD4 into wheat. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): (a) (b) Accomplishment: The AtPAD4 gene has been expressed in wheat cv. Bobwhite and in durum cv. Ben. Four independent AtPAD4 cv. Bobwhite lines and three cv. Ben lines have been identified. Impact: Homozygous progeny derived from these AtPAD4 expressing lines will be evaluated for FHB resistance in the greenhouse. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These AtPAD4 lines will not only provide germplasms that can be utilized in future breeding programs, but will also provide germplasms that can be utilized to study the involvement of this gene in wheat resistance to aphids, since PAD4 controls aphid resistance, as well. Accomplishment: AtNPR1 has now been expressed in durum cvs. Ben and Maier. Three homozygous lines in cv. Ben and two in cv. Maier are being propagated further. These five lines have shown the best level of enhanced FHB resistance compared to the nontransgenic cv. Ben and Maier plants. Impact: These germplasms will be made available to the Scab community later this year. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: (Form FPR07) 2

298 PI: Shah, Jyoti USDA-ARS Agreement #: The introduction of the Ubi1:AtNPR1 construct into durum cultivars is expected to provide new resistant germplasms for integration into durum breeding programs. (c) Accomplishment: Unlike two previous field trials in Kansas, in which AtNPR1 expressing cv. Bobwhite plants showed enhanced FHB resistance, no differences in FHB severity between the controls and the AtNPR1 transgenic plants was observed in a field trial conducted in spring 2007 in Minnesota. Impact: Additional trials are underway to study the conditions under which AtNPR1 conferred expression is most durable.. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Results from a series of field trials at different locations across the country will determine the conditions under which AtNPR1 expression provides good protection against FHB. (Form FPR07) 3

299 PI: Shah, Jyoti USDA-ARS Agreement #: Project 2: Signaling Mechanism in Host Defense and Susceptibility to Fusarium graminearum. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium graminearum is an important pathogen of wheat and barley, and other small grains. The development of strategies to control this pathogen has been further hampered by the lack of knowledge on host defense mechanism against the pathogen. We have utilized the model plant Arabidopsis thaliana to characterize plant mechanisms that contribute to defense and susceptibility to this pathogen. We have utilized a combination of genetic, molecular and biochemical approaches to identify phytohormones involved in resistance and susceptibility. These studies have indicated that salicylic acid and jasmonic acid signaling influence Arabidopsis defense against F. graminearum. We are simultaneously testing the role of these mechanisms in controlling FHB resistance in wheat, and using the Arabidopsis- F. graminearum interaction to identify additional genes important for host defense and susceptibility to this fungus. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): (a) Accomplishment: We have developed several transgenic wheat plants that express the bacterial NahG gene, which encodes salicylate hydroxylase that degrades salicylic acid. In addition, we have identified Arabidopsis WRKY18 as another gene that is a good candidate for enhancing FHB resistance in wheat and barley. In addition, we have demonstrated that a systemic acquired resistance (SAR) inducing activity purified from Arabidopsis also induces FHB resistance in wheat. Finally, our studies indicate that salicylic acid-regulated defenses target fungal spore germination. Impact: The NahG transgenic plants are being propagated further. Once homozygous lines are identified we will use these to study the role of salicylic acid in wheat defense against F. graminearum. A construct for expressing WRKY18 in wheat is being developed and this will be used to transform hexaploid and durum wheat to determine if it enhances FHB resistance in wheat. Our results indicate that studies in Arabidopsis can expedite the identification of plant genes and factors that can promote FHB resistance in wheat. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: NahG wheat lines have never been developed. These lines will provide an excellent genetic tool for the community to study plant defense against other pathogens, as well. (b) Accomplishment: We have uncovered a dual role for jasmonic acid in plant (Arabidopsis and wheat) interaction with F. graminearum. During the early stages of infection in wheat (first two days), jasmonic acid contributes to susceptibility by slowing down the activation of salicylic acid dependent defenses. In contrast, at later time points (4 dpi onwards), jasmonic acid enhances resistance, presumably by preventing spread of fungus. (Form FPR07) 4

300 PI: Shah, Jyoti USDA-ARS Agreement #: Impact: These results indicate that for high level FHB resistance in wheat, both salicylic acid and jasmonic acid signaling need to be targeted. We are working towards strategies to target both these mechanisms. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: These studies have uncovered a complex interaction between defense signaling mechanisms in wheat interaction with F. graminearum. Furthermore, these results indicate that in susceptible varieties the pathogen hijacks the plants jasmonic acid machinery to suppress the timely activation of defenses. Controlling this process in the host or the pathogens ability to do this, is key for developing good resistance to FHB. (Form FPR07) 5

301 PI: Shah, Jyoti USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Publications: Chaturvedi, R., Krothapalli, K., Makandar, R., Nandi, A., Sparks, A., Roth, M., Welti, R. and Shah, J. (2008) Plastid ω-3 desaturase-dependent accumulation of a systemic acquired resistance inducing activity in petiole exudates of Arabidopsis thaliana is independent of jasmonic acid. Plant J. 54: Makandar, R., Nalam, V., Essig, J.S., Schapaugh, M.A., Trick, H., Dill-Macky, R., and Shah, J. (2007) Enhancing Fusarium head blight resistance in wheat by manipulating mechanisms contributing to host resistance and susceptibility. In: Proceedings of the 2007 National Fusarium Head Blight Forum; Dec 2-4 ( Makandar, R., Chaturvedi, R., Sparks, A., and Shah, J. Fusarium graminearum macroconidia germination is inhibited by a salicylic acid and NPR1 (NONEXPRESSER OF PR GENES 1)- dependent mechanism that is constrained by jasmonate signaling in Arabidopsis thaliana. (submitted). Makandar, R., Nalam, V., Trick, H., and Shah, J. A complex interaction between salicylic acid and jasmonic acid controls wheat interaction with Fusarium graminearum, the causative agent of Fusarium head blight. (submitted). Presentations: 1. Title: Engineering Scab Resistance with Plant Defense Signaling Genes, at the National Fusarium Head Blight Forum, Kansas City, Missouri; December 2007 Presenter: Jyoti Shah 2. Title: Targeting Scab with Defense Regulatory Genes, at the 5th Canadian Workshop on Fusarium Head Blight, Winnipeg, Canada; November 2007; Presenter: Jyoti Shah 3. Title: Plants at War, Departmental of Biological Sciences Seminar, University of North Texas, Nov ; Presenter: Jyoti Shah (Form FPR07) 6

302 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Kevin Smith Institution: University of Minnesota Address: Department of Agronomy and Plant Genetics 411 Borlaug Hall 1991 Upper Buford Circle St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 154,654 Breeding and Genetics of Fusarium Head Blight Resistance in Barley. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGR Marker Assisted Selection for FHB Resistance in Barley. $17,581 HGG Developing Marker Information for Genetic Diversity and FHB Resistance in Barley. $ 53,171 VDUN Accelerated Development of Fusarium Resistant Barley Varieties. $ 83,902 Total Award Amount $ 154,654 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

303 PI: Smith, Kevin USDA-ARS Agreement #: Project 1: Marker Assisted Selection for FHB Resistance in Barley. 1. What major problem or issue is being resolved and how are you resolving it? Marker assisted selection (MAS) has only recently been adopted on a small scale by barley breeding programs for enhancing FHB resistance. We are working with the USDA Genotyping Center at Fargo to implement MAS with the best available markers to improve FHB resistance in barley. In the summer of 2007, we evaluated three breeding populations that were subjected to MAS for a chromosome 6H QTL in replicated FHB nurseries in Minnesota. In one of those three population, we saw a modestly significant effect of MAS (P=0.07) resulting in a 19% reduction in FHB severity. This QTL region (6H) has been inconsistently detected in the past in both mapping and MAS experiments. We are conducting additional fine mapping in this region and will initiate new MAS populations after we have better genetic information and more informative markers. This summer, we have three new populations that were subjected to MAS for the chromosome 2H bin 8 QTL planted in two FHB nurseries. Last fall we made 12 new crosses for the 2H QTL involving susceptible elite parents as well as new FHB variety candidates. The F2 generation is growing this summer and MAS will commence in the fall. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Tested MAS on three breeding populations with Chevron allele at 6H QTL. Impact: Variable effectiveness means that MAS using this QTL will need to wait until additional mapping work is complete. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: While the MAS results are not terribly impressive yet, we are advancing any promising lines in our breeding program. (Form FPR07) 2

304 PI: Smith, Kevin USDA-ARS Agreement #: Project 2: Developing Marker Information for Genetic Diversity and FHB Resistance in Barley. 1. What major problem or issue is being resolved and how are you resolving it? Progress in breeding for FHB resistance in barley to date has relied entirely on field-based phenotypic selection for disease severity and DON concentration of grain samples. Use of marker assisted selection (MAS) in early generations could provide a valuable tool to enrich breeding populations for resistance alleles at quantitative trait loci (QTL) prior to screening in the field. A major hurdle to using currently available QTL information for MAS is linkage of the resistance alleles to several undesirable traits (late heading, high grain protein concentration, tall plant height). Our project is focused on characterizing several important QTL regions in barley and resolving undesirable linkages. This information will lead to the implementation of MAS for FHB resistance in barley to complement ongoing traditional breeding efforts. We are currently involved in fine mapping three QTL regions (chromosomes 2H, 3H, and 6H). We are also using association mapping with breeding germplasm to identify new QTL that have not been discovered in traditional bi-parental mapping populations and that are presumably free of undesirable linkages due to selection in the breeding programs. This work is collaborative with the Busch Agriculture and North Dakota State University breeding and pathology programs and is primarily supported by the USDA Barley Coordinated Agricultural Project (CAP). This study is using data from 768 breeding lines evaluated in 5 trials for FHB and DON in 2006 and 2007 and screened with 3,000 SNP markers to identify QTL via genome-wide association mapping. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have generated large F2 populations for three QTL regions (2H, 3H, and 6H). We have used markers to select recombinants within the target region. We have planted recombinants from the 2H and 6H populations in FHB nurseries this summer. The data we collect this summer and in subsequent nurseries will help elucidate the position of FHB and DON resistance and help distinguish them from QTL for other linked traits. The 3H recombinants were evaluated in the greenhouse and we are still processing those samples and data. Preliminary data from the association mapping work indicates that there are ~8 QTL that were identified. All have relatively small effects with the largest having an R 2 value of We are currently investigating these QTL regions and developing MAS strategies that could be used to exploit them. Impact: Too early to assess the impact of these projects. (Form FPR07) 3

305 PI: Smith, Kevin USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We will soon be able to reveal the QTL that were detected in the association mapping study giving the Midwest barley breeders some potential new targets for MAS. Project 3: Accelerated Development of Fusarium Resistant Barley Varieties. 1. What major problem or issue is being resolved and how are you resolving it? None of the currently grown malting barley varieties are more resistant than the variety Robust which has dominated the barley acreage in the Midwest since We are conducting a comprehensive field-based breeding effort to develop new barley varieties with enhanced FHB resistance. In order for these new varieties to be adopted by growers they must also be agronomically competitive with current varieties and meet the quality standards of the malting and brewing industries. We conduct extensive field evaluation of FHB resistance in inoculated and mist-irrigated nurseries in three locations in Minnesota. Last year we evaluated over 12,000 plots for FHB resistance and submitted over 3,000 grain samples for DON analysis. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: One of our variety candidates, M122, was entered into AMBA plant-scale brewing evaluations. If this variety is rated satisfactory in its first year, we would anticipate releasing it as a variety in January M122 accumulates about half the level of DON compared to Robust. Two other variety candidates, M128 and M129, were rated satisfactory in their second year of AMBA pilot testing. These lines reduce DON by about 30%. We are currently evaluating their entrance into plant-scale brewing evaluation. This year we have four first-year entries into AMBA pilot-scale malt testing that all have enhanced FHB resistance. We have reached a point in our breeding program where the pipeline is full of FHB material and all our submissions to AMBA will have enhanced FHB resistance. Impact: The impact of this work will be felt after our first release with enhanced resistance to FHB. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: If M122 is approved by AMBA, this would be the first malting variety with enhanced resistance to M122 available to farmers in the Midwest. This line reduces DON by half compared to the currently grown varieties which is the current short term goal for the (Form FPR07) 4

306 PI: Smith, Kevin USDA-ARS Agreement #: USWBSI. In addition, M122 has been made available to other scientists that are evaluating integrated disease managements strategies to determine the effectiveness of combining genetic resistance with other FHB management strategies. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Nduulu L. M., A. Mesfin, G. J. Muehlbauer, and K. P. Smith Analysis of the chromosome 2(2H) region of barley associated with the correlated traits Fusarium head blight resistance and heading date. Theor. Appl. Genet. 115: Smith, K. P. The search for QTL to employ in marker assisted selection for FHB resistance in barley. Invited talk at the 5th Canadian Workshop on Fusarium Head Blight Nov Massman, J and K. P. Smith Prospects for Identifying Fusarium Head Blight Resistance QTL by Association Mapping using Breeding Germplasm. Proceedings of the 2007 National Fusarium Head Blight Forum, The Westin Crown Center, Kansas City, Missouri, Dec. 2-4, 2007, p Smith, K. P., E. Schiefelbein and G. Velasquez Development of Barley Variety Candidate M122 with Enhanced Resistance to FHB. Proceedings of the 2007 National Fusarium Head Blight Forum, The Westin Crown Center, Kansas City, Missouri, Dec. 2-4, 2007, p. 232 Massman, J. M. and K. P. Smith Investigating The Suitability Of Contemporary Breeding Populations For Association Mapping Of Fusarium Head Blight Resistance In Barley. Plant & Animal Genomes XVI Conference, January 12-16, 2008, Town & Country Convention Center, San Diego, CA. P329. (Form FPR07) 5

307 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Clay Sneller Institution: Ohio State University Address: Department of Horticulture and Crop Science OARDC 1680 Madison Ave Wooster, OH Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 88,608 Uniform Nursery for SRWW and Development Scab Resistance Varieties for Ohio. USWBSI Individual Project(s) USWBSI Research Area * HGG VDUN ARS Adjusted Award Project Title Amount Sources of Variation for DON Concentration in Moderately Resistant SRWW. $21,229 Uniform Nursery for SRWW and Development Scab Resistance Varieties for Ohio. $ 67,379 Total Award Amount $ 88,608 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

308 PI: Sneller, Clay USDA-ARS Agreement #: Project 1: Sources of Variation for DON Concentration in Moderately Resistant SRWW. 1. What major problem or issue is being resolved and how are you resolving it? Many lines with moderate resistance to FHB infection and/or spread still have high levels of DON. For some time researchers have speculated that there may be separate mechanisms of FHB resistance involving resistance to DON accumulation from either host degradation of DON or host inhibition of DON production. Thus a line with low FHB symptoms may still have high DON if is lacks these mechanisms. This has been a very difficult question to research as DON levels are often confounded with other FHB symptoms. No one has conducted a controlled study where FHB infection levels were controlled across many genotypes, degree of infection was assayed, and don data collected. We are conducting such a study to see if the amount of DON accumulate with increasing infection varies among SRWW genotypes. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We assayed heads from 80 SRWW cultivars. From each genotype we collected 15 heads with 0,1,2, or 3 infected spikelets (i.e. four sample per genotype, each with varying levels of infection based on visual symptoms). Each sample (80 x 4 =320 total) was assayed separately for DON and for degree of infection by using RT-PCR for Fusarium graminearum (Fg) and wheat specific primers. For the 4 samples from each genotype we regressed DON on Fg to assess the rate at which they accumulated DON with increasing infection. While we did not have replications in 2007, we did note large differences among the slopes of individual genotypes. Some genotype essentially did not increase DON despite increase FHB infection, while others showed dramatic increases in DON. Some genotypes had high DON despite relatively low infection levels. Impact: While unreplicated at this time, the data shows that genotypes do vary in the rate of DON accumulation with increasing infection. This provides preliminary evidence that there is genetic variation for DON accumulation. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: If repeated and verified, the results will be the first conclusive evidence for resistance to DON accumulation and provided sources for this type of resistance. If this is verified, then research will be needed to understand the mechanisms of this resistance (ie, host degradation of DON, or host inhibition of DON production), identify other sources, and improved methods to screen for this trait. (Form FPR07) 2

309 PI: Sneller, Clay USDA-ARS Agreement #: Project 2: Uniform Nursery for SRWW and Development Scab Resistance Varieties for Ohio. 1. What major problem or issue is being resolved and how are you resolving it? Planting varieties that are susceptible to FHB greatly increases the risk of yield loss and producing grain with excessive levels of DON. Host resistance is a primary mechanism for mitigating this risk. But increasing FHN resistance by itself is not enough as growers will not plant a variety that has other notable weaknesses such as moderate yield. FHB resistance must be combined with high yield, acceptable quality, and other traits. Our program aims to combine FHB resistance with other traits to produce SRWW varieties that are acceptable to Ohio growers. We use traditional breeding techniques supplemented with marker-assisted selection to accomplish the goal. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: 1. We screened 700 breeding lines from the OSU program, 160 lines from other programs, and 250 lines related to genetic studies. Of the OSU breeding lines, 57% had an FHB index < than the index of Freedom and 20% had an index < than that of Truman. 2. We screened 175 families from a cross of an FHB resistant line from CIMMYT crossed to a moderately susceptible SRWW. Significant segregation was noted with several lines having Truman-like resistance. We developed a mapping population from the families for further phenotyping in We used MAS to continuing BCing multiple QTL into 27 different genetic backgrounds. Of these 21 were advanced to the BC1F1 by the end of the grant, 3 to t he BC3F1 and 3 to the BC3F2 generation. We also used MAS for F1 enrichment of F1 from three-way crosses where the new BC derivatives were used as parents. 4. We coordinated the PNUWWSN and NUWWSN. For the 2007 nurseries this involved gathering and analyzing data from 13 cooperators on evaluation of 104 SRWW lines and checks. The seed of 120 lines for the 2008 test were distributed to 16 cooperators in three countries. Impact: 1. Through the screening we identified many SRWW with at least moderate resistance to FHB. We released three of these based on their FHB resistance and overall performance. One of these, Malaabr (OH ) has Truman-like resistance, while Bromfield (OH ) and OH (branded) have Freedom-like resistance. 2. The segregation and the resistance of the progeny of the CIMMYT/SRWW cross indicates that the CIMMYT line is contributing genes for resistance and we postulate these are novel to SRWW. 3. The sizable BCing effort will develop a broad base of SRWW with known genes for FHB resistance. All the RP have been selected to have acceptable; agronomic value (Form FPR07) 1

310 PI: Sneller, Clay USDA-ARS Agreement #: based on current tests as well as moderate FHB resistance. By adding new FHB QTL to these genetic backgrounds we hope to attain strong FHB resistance in argonomically useful backgrounds. 4. The successful execution of the uniform FHB nurseries ensures reliable FHB ratings of germplasm from 13 public and private breeding programs. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The release of Malabar, Bromfield, and OH provide Ohio growers high yield varieties with good FHB resistance. This expands their options tremendously, especially access to a variety with Malabar with Truman-like resistance and excellent yield potential for Ohio. Attaining acceptable FHB resistance requires pyramiding several novel FHB resistance alleles. The CIMMYT line may have some novel QTL, especially from the D genome and this would provide breeders the ability to make and test more gene combinations than in the past. Few agronomically competitive SRWW lines have major FHB QTL and this makes breeding difficult as we do not know what genes our resistant parents have and often must use agronomically unfit parents as sources of these QTL. The collection of breeding lines with FHB QTL that we are generating will significantly improve the agronomic value and diversity of lines with FHB QTL as well as potentially lead directly to release of an improved cultivar. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Sneller, C, P. Paul, L. Herald, B. Sugerman, and A. Johnston Report on the Northern Uniform Winter Wheat Nurseries. in Proceeding of the 2007 National Fusarium Head Blight (NFHB) Forum, Kansas City, MO, 12/2-4/2007, pages Sneller, C.H, P. Paul, and M. Guttieri Prevalence of resistance to Fusarium Head Blight in Eastern US sort red winter wheat. Submitted to Crop Sci. (Form FPR07) 2

311 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Mark Sorrells Institution: Cornell University Address: Department of Plant Breeding 252 Emerson Hall Ithaca, NY Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 35,600 Development of Fusarium Head Blight Resistant Wheat Varieties - Cornell. USWBSI Individual Project(s) USWBSI Research Area * VDUN ARS Adjusted Award Project Title Amount Fusarium Head Blight Resistant Soft White Wheat Variety $35,600 Development for the Northeastern US. Total Award Amount $ 35, Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

312 PI: Sorrells, Mark USDA-ARS Agreement #: Project 1: Fusarium Head Blight Resistant Soft White Wheat Variety Development for the Northeastern US. 1. What major problem or issue is being resolved and how are you resolving it? Field evaluation methods continue to improve and have generated useful results for the past 4 years thus eliminating the problem of low infections. In addition, the evaluation protocol was an issue, which we resolved by consulting with other collaborators. The revised scoring system is similar to the method recommended on the USWBSI web site and seems to be fairly reliable but not quite as accurate as the total counts on 1 meter rows that we used previously. We also continued to refine a protocol for selecting single plants in early generations. We can now reliably identify resistant genotypes about 40-50% of the time based on two years of evaluation of the progeny of those single plants. In two independent experiments, 38 and 48% of the putative resistant single plants were resistant when evaluated in replicated 1 meter rows. Lodging and diseases diseases were not a problem this year. We evaluated a mass inoculation method for nurseries that could not be placed under the mist irrigation. Infection rates were much lower but we could still identify susceptible genotypes. For 2009, we are going to continue with this mass inoculation of our main breeding nursery to eliminate susceptible genotypes before yield evaluation. In our marker assisted selection program we added the screening of our recurrent parents for the 5AS resistance gene and found that nearly all were polymorphic with our two sources OH and OH These have been crossed to all of our recurrent parents, including E0028, NY , NY , NY , Jensen, P25W41, CalResel-L. We have also crossed our two Caledonia derived FHB lines to E0028 and CalResel-L. These will all be advanced again in our winter greenhouse. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Jensen soft white winter wheat went into production this past year. It is moderately resistant to fusarium head blight and preharvest sprouting, the two main problems in this growing region. CaledoniaReselect-L which also shows excellent resistance to FHB was approved for release and foundation seed production began this year. These are the first FHB resistant soft white winter wheat varieties in the northeastern region and represent an important milestone for our growers. Impact: The importance of these white wheat varieties is enormous for this growing region. White wheat is a very important raw material for the whole grain products produced by companies in this region. In recent years there has been a shift in acreage from white to red wheat thus creating a severe shortage. Buyers have been offering a premium for farmers who grow a quality crop of white wheat. (Form FPR07) 2

313 PI: Sorrells, Mark USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The resistance in these varieties is non-chinese which can be combined with Chinese sources to increase resistance to FHB. These two new varieties provide northeastern farmers a choice of white wheat varieties with high yield, and good agronomic traits that will produce a more reliable crop. We anticipate that this will help to stabilize white wheat production in this region. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Presentations: 6/5 Aurora, NY Small grains management field day Jensen Soft White Winter Wheat with Fusarium Head Blight Resistance for New York 6/19 WSMI radio interview Varieties and resources for farmers growing wheat who want to avoid head scab. 7/8 Ithaca, NY Seed grower s field day Cornell Wheat Breeding Program and Jensen Soft White Winter Wheat with Fusarium Head Blight Resistance for New York No publications that have a journal format citation. (Form FPR07) 3

314 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Brian Steffenson Institution: University of Minnesota Address: Department of Plant Pathology 495 Borlaug Hall 1991 Upper Buford Circle St. Paul, MN Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 49,638 Evaluation of Hordeum Germplasm for Resistance to Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGR Accelerated Screening of Hordeum Germplasm for FHB Resistance. $33,540 IIR Development of Digital Disease Diagrams for Assessing FHB of Barley. $ 16,098 Total Award Amount $ 49,638 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

315 PI: Steffenson, Brian USDA-ARS Agreement #: Project 1: Accelerated Screening of Hordeum Germplasm for FHB Resistance. 1. What major problem or issue is being resolved and how are you resolving it? FHB threatens the existence of the barley industry in the Upper Midwest. Deployment of resistant cultivars is the most effective and environmentally sound means of managing this disease; however, sources with high levels of resistance to Fusarium graminearum and its toxins are lacking. Thus, the primary objective of this research is to identify Hordeum germplasm with the highest level of FHB resistance possible. Our specific activities also involve the sourcing of unique Hordeum germplasm from foreign genebanks. Our USWBSI research program is an ongoing effort to systematically evaluate unique Hordeum germplasm from USDA and foreign genebanks for resistance to FHB in the Upper Midwest and also off-season nurseries in China. The screening of the entire six-rowed spring and winter barley collection from the USDA National Small Grains Collection (NSGC) is complete. Additionally, we have evaluated over 1,233 wild barley (Hordeum vulgare subsp. spontaneum) accessions from the NSGC. We have essentially completed evaluations of six-rowed and wild barley germplasm from the NSGC, but are also sourcing unique Hordeum accessions from other gene banks around the world, including the N. I. Vavilov All-Russian Scientific Research Institute of Plant Industry (VIR) in St. Petersburg, Russia, the Station federale de recherches en production vegetale de Changins (SFRSPP) in Nyon, Switzerland, the Nordic Gene Bank (NGB) in Alnarp, Sweden, the Institute for Cereal Crops Improvement (ICCI) in Tel Aviv, Israel, Plant Genetic Resources of Canada (PGRC), Saskatoon, Canada, and International Center for Agriculture in the Dry Areas (ICARDA), Aleppo, Syria. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Additional sources of resistance to FHB are needed in breeding programs. We have procured nearly 11,070 additional accessions from various genebanks and evaluated them in FHB nurseries in the Upper Midwest and/or China. From these evaluations, we have identified accessions with resistance levels comparable to the resistant six-rowed check Chevron. Five hundred and ninety-four of these accessions came from the NSGC, 43 from the VIR, 11 from the SFRSPP, 3 from the NGB, 3 from the ICCI, 9 from the PGRC, and 0 from ICARDA. Impact: We have discovered new and diverse sources of FHB resistance in barley that are likely different from those already reported. The identification of resistant germplasm is the first step in developing barley cultivars with enhanced resistance to FHB and the accumulation of toxins. Accessions identified as resistant in our initial screening nurseries are immediately distributed to barley breeders for crossing within their FHB resistance-breeding program. In FY07, we (Form FPR07) 2

316 PI: Steffenson, Brian USDA-ARS Agreement #: provided four more sources of FHB resistance to barley breeders. These sources have already been crossed to adapted material and are now in the breeding pipeline. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Our program provides breeders with the germplasm needed to pyramid additional FHB resistance loci into their lines and enhance resistance to the pathogen. Introgression of resistance alleles from these sources into advanced barley cultivars will reduce the detrimental impact of the FHB pathogen and its associated toxins. In the Minnesota and North Dakota barley breeding programs, several advanced lines with partial resistance to FHB are in pilot and plant scale testing. These advanced lines were bred with sources of partial FHB resistance identified from our screening of NSGC germplasm. These lines, pending American Malting Barley Association approval, may be available to growers within the next few years. (Form FPR07) 3

317 PI: Steffenson, Brian USDA-ARS Agreement #: Project 2: Development of Digital Disease Diagrams for Assessing FHB of Barley. 1. What major problem or issue is being resolved and how are you resolving it? Many projects funded by the USWBSI on barley require a reliable and efficient method for assessing FHB severity. This includes studies of resistance in raw germplasm accessions, breeding lines, and segregating populations; effectiveness of various residue management treatments in reducing pathogen inoculum; efficacy of chemicals and biologicals in controlling FHB; and various epidemiological parameters affecting disease spread. In the barley improvement programs, the time required for disease assessment is a serious bottleneck as over 12,000 lines are evaluated each summer--often times in the span of just a few day s time. To reduce the time required for assessing FHB in barley, we initiated a project to develop prototype disease diagrams based on digital images of actual infected spikes from the field. Archetypal healthy and diseased spikelets were selected and digitally cut from these spike images, and then used as building blocks to create idealized two-rowed and six-rowed spikes exhibiting various disease severities. Prototypes of fourteen standard digital spike images exhibiting disease severities from 1-100% were developed for both two-rowed and six-rowed barley. The objectives of this study were to 1) develop a refined and comprehensive set of digital disease images for estimating FHB severity in both six-rowed and two-rowed barley, and 2) assess the accuracy, reproducibility, and efficiency of using these disease images for estimating FHB severity in various applications. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We developed and refined a set of 14 standard digital spike images exhibiting disease severities from 1-100% for both two-rowed and six-rowed barley. Twelve novice, intermediate, and experienced FHB researchers utilized the digital diagrams to assess its accuracy and efficiency for rating FHB severity in barley spikes collected from the field. All raters under-estimated the true severity level of six-rowed spikes at disease levels of 35% and above. Using the digital images, the savings in rating time ranged from 37 seconds to 12 minutes, 35 seconds. With the two-rowed spikes, all raters under-estimated the true severity level at disease levels of 25% and above. The use of the rating scale did not result in a time savings in all cases; in fact, some raters required 3 minutes longer to complete the assessments using the rating scale over performing actual diseased kernel counts. However, other raters saved up to 10 minutes using the digital images. Impact: To our knowledge, this is the first set of digital images developed to assess FHB severity in both two-rowed and six-rowed barley. The digital images may provide a more efficient means of evaluating barley for FHB severity under some applications, such as the disease resistance evaluations. (Form FPR07) 4

318 PI: Steffenson, Brian USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The digital disease images developed and validated for rating FHB severity in two- and sixrowed barley will provide a sufficiently accurate assessment of disease, but with greater efficiency (i.e. accuracy plus speed) than the spikelet count method. This will be a tremendous benefit to the breeding programs where thousands of lines need to be evaluated within a very short period of time. (Form FPR07) 5

319 PI: Steffenson, Brian USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. None. (Form FPR07) 6

320 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Jeffrey Stein Institution: South Dakota State University Address: Plant Science Department Box 2108 Brookings, SD Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement FHB Epidemiology on Spring Wheat in South Dakota. Title: FY07 ARS Award Amount: $ 96,585 USWBSI Individual Project(s) USWBSI Research Area * CBCC EEDF EEDF ARS Adjusted Award Project Title Amount Influence of Residue Management on Inoculum Production, Disease, and DON. $19,512 Environmental Factors Influencing FHB and DON Accumulation in Hard Red Wheat. $ 38,537 Environmental Factors Influencing FHB and DON Accumulation in Malting Barley. $ 38,536 Total Award Amount $ 96,585 7/14/08 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

321 PI: Stein, Jeffrey USDA-ARS Agreement #: Project 1: Influence of Residue Management on Inoculum Production, Disease, and DON. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) continues to be a serious problem in the U.S. and Canada. The most effective management of this disease occurs when an integrated approach is taken that combines a resistant variety, appropriately timed fungicide application, and residue management practices that limit fungal survival and/or inoculum production. The first two strategies have been investigated thoroughly and we now have partially resistant varieties and moderately effective fungicide programs. In contrast, residue management has essentially been limited to rotational or tillage practices. Unfortunately, growers are often limited in what schemes they can do for a variety of reasons. If additional practices were available for residue management, the frequency and severity of FHB epidemics and/or deoxynivalenol (DON) contamination in grain might be effectively reduced. The objectives of the proposed research were to utilize A) physical residue management and B) biological control agents in an attempt to inhibit the growth, sporulation, and infection of wheat by Gibberella zeae. To address these objectives we planted spring wheat into plots containing maize residue that had been left to over-winter after a standard harvest, shredded further, or incorporated into the soil. A biocontrol agent or fungicide was applied to each plot in the fall or spring. The wheat was un-inoculated and we recorded the disease severity 18 days after heading and determined the final DON accumulation in the grain for individual plots. We also conducted assays designed to determine the activity of saprophytic microbes in the residue. These organisms are potential antagonists to G. zeae and could impact sporulation and infection. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Unfortunately for this project, the environmental conditions leading up to, and during, the period of heading in 2007 were highly unfavorable for spore production and infection by G. zeae. We had essentially no precipitation between early June and late July in Brookings and this negatively impacted our ability to compare both the residue processing and biological treatments. Similarly, the soil and surface residues in field plots were very dry and CO 2 emissions were detected but quite low, making it impossible to compare treatments. When residue samples were processed in the lab using FDA hydrolysis, most of the samples were found to have a fairly similar level of saprophytic activity. The only treatment that had an elevated level of activity was the fall-applied Trichoderma. This organism is both mycoparasitic and saprophytic and could be impacting the ability of G. zeae to survive in the residue. It was also noted that the physical processing of maize residues in the fall significantly reduced the amount of material that was present in the spring. In fact, it was difficult to visual discern the tillage and fall-residue shredding treatments. (Form FPR07) 2

322 PI: Stein, Jeffrey USDA-ARS Agreement #: Impact: None to date. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The aggressive chopping of maize residues following harvest in the fall results in the accelerated decomposition of this material with most of it degrading by the next growing season. Although we have yet to specifically document any impacts from this treatment on FHB, we have found a strong relationship in other studies between the level of maize residue present, the number of spores reaching wheat heads in that field, and the resulting disease severity and mycotoxin accumulation. Therefore, it stands to reason that the processing of cereal residues following harvest will reduce both the level of in-field inoculum and the risk of economic losses from FHB. (Form FPR07) 3

323 PI: Stein, Jeffrey USDA-ARS Agreement #: Project 2: Environmental Factors Influencing FHB and DON Accumulation in Hard Red Wheat. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) continues to be a serious problem in the U.S. and Canada. The most effective management of this disease occurs when an integrated approach is taken that combines a resistant variety, appropriately timed fungicide application, and the management of in-field inoculum. Previous research funded by the U.S. Wheat and Barley Scab Initiative has resulted in the development of predictive models for wheat that target infection at a specific time point in the crop developmental stage (i.e. early flowering) using weather data collected over the previous 7 days. Unfortunately, this system has several limitations and does not model deoxynivalenol (DON) accumulation in the grain. Thus, the primary objective of this project was to conduct research that would facilitate the development of mechanistic models that attempt to predict both disease and DON accumulation using weather, varietal resistance, and timing of infection. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: Unfortunately for this project, the environmental conditions leading up to, and during, the period of heading in 2007 were highly unfavorable for infection by G. zeae. We had essentially no precipitation between early June and late July in Brookings and this negatively impacted our ability to compare the interaction between varietal resistance, timing of infection, and environment. That stated, the early (or optimal) inoculation timing had significantly higher disease incidence and severity, and lower yield, than the non-inoculated and late-inoculate treatments. In comparison, there were no significant differences between the treatments with respect to DON concentration. Impact: None to date. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Nothing to date. (Form FPR07) 4

324 PI: Stein, Jeffrey USDA-ARS Agreement #: Project 3: Environmental Factors Influencing FHB and DON Accumulation in Malting Barley. 1. What major problem or issue is being resolved and how are you resolving it? Fusarium head blight (FHB) of barley continues to be a serious problem for producers in the Northern Great Plains. Barley production in the Dakotas and Minnesota has declined steadily since the early 1990 s and this can be attributed to, at least in part, the re-emergence of Fusarium head blight. Of particular importance to barley production is the accumulation of deoxynivalenol (DON) in the grain. Significant progress has been made in recent years in the development of disease forecast models for wheat; however, the effectiveness of these models in predicting disease, and more importantly DON accumulation, for barley is questionable. We are addressing this issue by attempting to develop models for eventual incorporation into forecasting systems which can predict disease and DON accumulation for barley. Such a system would offer producers the information required to make effective management decisions. The objectives of the proposed research were to 1) continue developing a database of information on environment, crop residue, field disease, and mycotoxin levels for spring barley in the region and 2) begin identifying variables that are predictive of disease and DON accumulation in barley in order to develop models and risk advisory systems. Objective 1 was conducted in collaboration with researchers at North Dakota State University and the University of Minnesota. Plots were planted at 12 locations throughout the region and the environment was monitored at each location during the growing season. Field ratings of disease were taken and DON concentration in the grain was quantified. For Objective 2, approximately 100 variables, both simple and complex, were generated using environmental parameters that are known to impact the biology of this pathosystem (temperature, relative humidity, etc). Correlation analysis was then conducted on the combined data sets to determine which, if any, of these factors were strongly associated with field disease or DON content in the grain. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The most significant accomplishment of this project was the continued development of a database of information on weather, crop residue, field disease, and mycotoxin levels for spring barley. We now have 30 location-years in this dataset with information on both 2- and 6-row barley cultivars commonly grown in the region. The disease severity and DON concentrations range from 0-59% and ppm, respectively. It should be noted that many of the locations had very little disease and mycotoxin in 2007 due to dry conditions during the period of heading. Therefore, the total dataset is still insufficient for full model development. (Form FPR07) 5

325 PI: Stein, Jeffrey USDA-ARS Agreement #: Impact: To date, this effort has not directly impacted barley producers in the region. However, a future impact is expected following the model development effort. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Currently, we lack the number of location-years required for the development of accurate models for disease and DON prediction. However, as the dataset being generated by this project increases in size we will be able to better understand which environmental factors are best able to predict disease and DON in barley. This will lead to the development of models that are robust and effective at predicting FHB and DON accumulation in barley. (Form FPR07) 6

326 PI: Stein, Jeffrey USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Peer-reviewed articles: Paul, P., P. Lipps, G. Shaner, G. Buechley, T. Adhikari, S. Ali, J. Stein, L. Osborne, and L. Madden A distributed-lag analysis of the relationship between Gibberella zeae inoculum density on wheat spikes and weather. Phytopath. 97: Osborne, L.E., and J.M. Stein Epidemiology of Fusarium Head Blight on Small Grain Cereals. Intl. J. Food. Microbiol. 119: Non-peer reviewed (reports, posters, presentations, and related): Osborne, L Wheat and Barley Scab Risk Advisory. (17 updates). Disease forecast and commentary. URL: Osborne, L The Fusarium Head Blight Prediction Center. (17 commentaries). Sitespecific weather and disease forecast information. URL: Nita, M., E. DeWolf, L. Madden, P. Paul, G. Shaner, T. Adhikari, S. Ali, J. Stein, and L. Osborne Integrated management of Fusarium Head Blight (FHB) and deoxynivalenol concentration. Presented at the Joint Meeting of the APS/SON. San Diego, CA. Phytopathology 97:S85. Stein, J.M., L.E. Osborne, S. Neate, and C. Hollingsworth Environmental Factors Influencing FHB Severity and DON in Barley. Poster: Proceedings of the 2007 National Fusarium Head Blight Forum, Kansas City, MO. Canty, S.M., Clark, A., Ellis, D., and Van Sanford, D. (Eds.), University of Kentucky, Erlanger, KY. pp Osborne, L.E., J.M. Stein, K.D. Glover, and C.A. Nelson Spore Load, Disease, and DON: An Inoculum Gradient Study Using Sister Wheat Lines. Poster: Proceedings of the 2007 National Fusarium Head Blight Forum, as before. pp Osborne, L.E., J.M. Stein, and C.A. Nelson Spore Load, Disease, and DON: A Four Year Variety by Residue Study for FHB Management. Poster and Report: Proceedings of the 2007 National Fusarium Head Blight Forum, as before. pp Nita, M., E. DeWolf, L. Madden, P. Paul, G. Shaner, T. Adhikari, S. Ali, J. Stein, L. Osborne, and S. Wegolu Mechanistic Simulation Models for Fusarium Head Blight and Deoxynivalenol. Poster: Proceedings of the 2007 National Fusarium Head Blight Forum, as before. pp (Form FPR07) 7

327 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Laura Sweets Institution: University of Missouri Address: Department of Plant Microbiology and Pathology 210 Waters Hall Columbia, MO Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 14,634 Uniform Trial to Evaluate Efficacy of Fungicides and Biologicals Against Fusarium Head Blight. USWBSI Individual Project(s) USWBSI Research Area * CBCC CBCC ARS Adjusted Award Project Title Amount Evaluation of Integrated Management Strategies for Fusarium Head Blight. $9,756 Uniform Trial to Evaluate Efficicay of Fungicides and Biologicals against Scab. $ 4,878 Total Award Amount $ 14,634 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

328 PI: Sweets, Laura USDA-ARS Agreement #: Project 1: Evaluation of Integrated Management Strategies for Fusarium Head Blight. 1. What major problem or issue is being resolved and how are you resolving it? The severity of Fusarium head blight (FHB) epidemics in the United States has caused enormous yield and quality losses in both wheat and barley over the last decade. The development of this disease is dependent on host genetics, a range of favorable environmental conditions, the prevalence of the causal fungus and the survival and spread of the causal fungus. Control of this disease has been difficult because of the complex nature of the host/pathogen interaction. Management of FHB and the associated mycotoxin DON have not been achieved by any single control measure. An integrated approach is critical to attaining the best possible control of FHB in any given environment. As a result of a workshop sponsored by the Chemical, Biological and Cultural Control Research Area in 2006, a protocol for a multi-state project focusing on integrated management strategies for FHB was developed. The research portion of this project would involve multi-state trials evaluating crop sequence, variety selection and fungicide application as an integrated management program for FHB. Timely dissemination of the research results is also a priority of this project. The University of Missouri cooperated in this multi-state project following the protocol developed by a subcommittee from the workshop participants during the season. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The data from the Missouri trial does indicate a significant difference in DON levels with varieties and with crop sequence. As expected more susceptible varieties tended to have higher levels of scabby kernels and higher DON levels than varieties with more resistance to FHB. However, all varieties had lower DON levels when planted in soybean residue rather than corn residue. Fungicide application did not seem to have an impact on yield or DON level regardless of variety or crop sequence. The plan is to repeat the trial in to see if results are reproducible and to obtain a second year s data on the integrated management of FHB. Impact: The purpose of the Integrated Management Strategies for Fusarium Head Blight Project is to provide growers with data on the value of utilizing multiple management options to reduce both FHB and DON levels in grain crops. Compilation of data from similar trials from all locations provides valuable information on which practices and/or combination of practices may reduce the incidence and severity of FHB and may then impact the DON levels in harvested grain. For Missouri one key point would be the value of crop sequence or rotation in reducing FHB and DON in winter wheat. (Form FPR07) 2

329 PI: Sweets, Laura USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The uniform trial data is compiled across all locations participating in the trial and this information in presented at the National Scab Forum, is published in the National Scab Forum proceedings and is available through the U.S. Wheat and Barley Scab Initiative web site as well as through individual state reports and web sites. Growers and agribusiness personnel in Missouri and academic and agricultural professionals throughout the United States have access to this information. Again, from the first year of this study one important trend was the lower incidence of FHB and lower DON levels in wheat planted into soybean residue as compared to wheat planted into corn residue. The importance of crop rotation in managing FHB may have been overlooked in recent years. (Form FPR07) 3

330 PI: Sweets, Laura USDA-ARS Agreement #: Project 2: Uniform Trial to Evaluate Efficicay of Fungicides and Biologicals against Scab. 1. What major problem or issue is being resolved and how are you resolving it? Scab or Fusarium head blight (FHB) continues to be a problem on soft red winter wheat grown in Missouri. Although a state wide epidemic has not occurred for several years, each year there are areas within the state that have weather conditions favorable for disease development as the winter wheat crop is flowering. Producers in these areas see a direct impact from yield reduction and may see an indirect impact due to DON levels or quality issues when the grain is marketed. High levels of FHB in the crop or of DON in the grain also cause significant problems for elevators accepting the grain and processors trying to use the grain. One management option would be the use of fungicides or biological control agents to minimize FHB infection. The Uniform Scab Fungicide Trial was set up to identify safe fungicides that are effective against FHB. A second uniform trial to evaluate the efficacy of biological control agents against FHB has also been established. Given sets of fungicides and biological control agents are evaluated for consistency of performance across a wide range of wheat classes and varieties, barley classes and environments. For the fungicide portion of the uniform trial, the emphasis is on new fungicide chemistries, new combinations of products, more precise application timing and application techniques. In the biologicals portion of the uniform trial, the emphasis has shifted to biologicals available in formulations that would be practical for on-farm use. The identification of safe fungicides or the development of safe, easy to use biological control agents which effectively control FHB would benefit producers, agribusinesses and ultimately consumers. The set of fungicides and biological control agents for the Uniform Scab Trials were evaluated on two soft red winter wheat varieties in Missouri during the season. The field work for this trial has been completed. Samples were submitted for DON analysis and data was analyzed for the annual trial report. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: The Uniform Trials to evaluate efficacy of fungicides and biological control agents against FHB were conducted in Missouri during the season. Planting went well and the stands were good last fall. During the winter months there were two major ice storms which covered both trials with about two inches of ice. Weather was also a major factor from March to early April. The last three weeks of March temperatures were unusually warm resulting in the second warmest three weeks in March since Then from April 4 to April 7, the temperatures plummeted and record lows were set for most of those days. The abrupt change from three weeks of very warm weather which encouraged wheat green up and growth to night time temperatures of 17 or 18 degrees F resulted in extensive low temperature damage to wheat. Damage ranged from plants that were killed by low temperatures to plants that had stem damage especially to the main stem but managed to (Form FPR07) 4

331 PI: Sweets, Laura USDA-ARS Agreement #: survive to plants that had poor head development and fill to plants that showed little or no negative effects from the low temperatures. The Uniform Fungicide Trial was conducted on the two varieties Roane and Truman. Roane sustained moderated low temperature damage and Truman sustained little low temperature damage. In the Uniform Biological Trial the varieties were Elkhart and Roane. Again Roane did not show major damage from the low temperature but the Elkhart was severely affected. In the Elkhart stands were thin and uneven. Weather conditions during flowering were conducive to the development of FHB. Fungicide treatments were applied at the appropriate timings. Biological control agents were applied under ideal conditions for growth of the biological control agents and for development of FHB. This year all of the biologicals were supplied in sufficient quantity so no additional increase of biological inoculum was necessary. Furthermore all of the biological control agents were applied with ease; the formulations didn t clog spray nozzles or cause any other application problems Unfortunately, weather conditions from flowering on were not favorable for the development of FHB. Both incidence and severity of FHB were low when those ratings were taken in the field. Scabby kernel counts were also low. All three varieties did have more blank kernels and shriveled or shrunken kernels than normal. It is difficult to distinguish between kernels damaged by low temperatures and kernels with scab in visual ratings. The DON levels were mostly below detectable levels for all treatments in both trials indicating that kernel damage was more likely the result of adverse environmental conditions than to FHB. But the strength of the Uniform Fungicide and Biological Trials has been and continues to be in the replication of the same sets of treatments across a number of locations. Although results in one location in a given year may not be statistically significant, across all locations there should be significant results that provide valuable information for all wheat and barley producing areas of the U. S. Impact: The purpose of the Uniform Fungicide Trial and the Uniform Biological Control Agent Trial is to provide growers with data on the efficacy of these materials that will help them in making management decisions related to FHB. Compilation of uniform trial data from all locations provides valuable information on which fungicides or biological control agents reduce the incidence and/or severity of FHB, which may impact the DON levels in harvested grain and application timings and techniques that provide the best results. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: The uniform trial data is compiled across all locations participating in the trials and this information in presented at the National Scab Forum, is published in the National Scab Forum proceedings and is available through the U.S. Wheat and Barley Scab Initiative web site. Growers and agribusiness personnel in Missouri as well and academic and agricultural professionals throughout the United States have access to this information. (Form FPR07) 5

332 PI: Sweets, Laura USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Presentations: Crop Injury Diagnostic Clinic, Field Crop Disease Session, July 2007, Columbia, MO Crop Management Conference, Field Crop Disease Update, December 2007, Columbia, MO Pesticide Applicator Training Recertification Sessions, Field Crop Disease Update, January 2008, multiple locations throughout Missouri Wheat Diseases, MFA Training, February 2008, Columbia, MO Training Session for Missouri Seed Improvement Association Wheat Inspectors, June 2007, Columbia, MO Wheat Tour, May 2008, Lamar, MO Teleconference phone calls with Extension field staff during the spring and summer of 2007 Proceedings: Paul, P., L. Madden, M. McMullen, D. Hershman, L. Sweets, S. Wegulo W. Bockus, S. Halley and K. Ruden An integrated approach to managing FHB and DON in wheat: uniform trials Proceedings of the 2007 National Fusarium Head Blight Forum. December 2-4, Kansas City, Missouri. pp Paul, P., L. Madden, M. McMullen, D. Hershman, D. Brown-Rytlewski, L. Sweets, E. Adee, C. Bradley, B. Padgett and K. Rudeen Fungicide effects on FHB and DON in wheat across multiple locations and wheat classes: uniform fungicide trials Proceedings of the 2007 National Fusarium Head Blight Forum. December 2-4, Kansas City, Missouri. pp Yuen, G. L., C.C. Jochum, K. R. Ruden, J. Morgan, B. H. Bleakley and L. E. Sweets Results from the 2007 standardized evaluation of biological agents for the control of Fusarium head blight on wheat and barley. Proceedings of the 2007 National Fusarium Head Blight Forum. December 2-4, Kansas City, Missouri. pp Extension Publications: Manuals: Bradley, K. W., L. E. Sweets, W. C. Bailey, J. A. Kendig, and J. A. Wrather Missouri pest management guide. University of Missouri Extension publication M171. (Form FPR07) 6

333 PI: Sweets, Laura USDA-ARS Agreement #: Newsletter Articles: Sweets, L. E. Foliage diseases of winter wheat and their management. Integrated Pest and Crop Management Newsletter 17(7). Sweets, L. E. Foliar fungicides labeled for use on winter wheat. Integrated Pest and Crop Management Newsletter 17(7). Sweets, L. E. Wheat diseases. Integrated Pest and Crop Management Newsletter 17(8). Sweets, L. E. Fusarium head blight or scab of wheat. Integrated Pest and Crop Management Newsletter 17(9). Sweets, L. E. Evaluate winter wheat seed quality prior to planting. Integrated Pest and Crop Management Newsletter 17(13). Sweets, L. E. Seed treatment fungicides labeled for use on winter wheat. Integrated Pest and Crop Management Newsletter 17(13). (Form FPR07) 7

334 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Frances Trail Institution: Michigan State University Address: Department of Botany & Plant Pathology 342 Plant Biology Lab East Lansing, MI Phone: Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 59,424 Starch Degradation by Gibberella zeae and its Role in Fueling Development. USWBSI Individual Project(s) USWBSI Research Area * Project Title ARS Adjusted Award Amount HGG Understanding Vascular Gels as a Resistance Response. $15,522 PGG Survival of Fusarium graminearum in Crop Debris and the Role of Protective Antimicrobials. $ 43,902 Total Award Amount $ 59,424 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

335 PI: Trail, Frances USDA-ARS Agreement #: Project 1: Understanding Vascular Gels as a Resistance Response. 1. What major problem or issue is being resolved and how are you resolving it? Resources for scab resistance are quite limited. We observed that about one third of the Norm plants we inoculated in the greenhouse had limited spread (Type 2 resistance) due to the presence of pectic gels in the xylem. We performed a progeny test on those plants that showed the resistance. The same percentage of progeny had the reaction as in the parental line. This shows that the reaction is a consistent trait within the population, suggesting this is not due to heterozygosity in the parent line for this trait. The reaction is likely to be an induced response, perhaps the frequency is due to the rapidity of the response in some individuals. We are now working with Janet Lewis, testing this response in other varieties. We see it in resistant varieties in a higher frequency. It is important to note that this response does not kill the upper half of the wheat head. It appears to act just on the floret which is infected. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have shown that the pectic gels formed consistently in the progeny of Norm, indicating this form of resistance may be uniformly expressed in Norm. Impact: Types of resistance to scab are limited in wheat. If this resistance mechanism can be bred into lines, and enhanced, it may be very effective in limiting disease spread. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We have identified a novel resistance mechanism that may be useful for breeding into lines of wheat. (Form FPR07) 2

336 PI: Trail, Frances USDA-ARS Agreement #: Project 2: Survival of Fusarium graminearum in Crop Debris and the Role of Protective Antimicrobials. 1. What major problem or issue is being resolved and how are you resolving it? We have been investigating the formation of inoculum that arises out of the crop debris. We investigated the role of relative humidity in the formation of perithecia on crop debris (we used colonized stems that had been inoculated at the wheat head). We show that exposure to less that 70% humidity for 5 days in stems harvested at grain maturity minimized mycelium growth and maximized perithecium production when humidity subsequently rose to above 70%. On the other hand, exposure to 100% humidity with no exposure to lower humidity resulted in massive mycelial growth and perithecium production. The data suggest that when optimal conditions for harvest (low humidity) are present during grain harvesting, that the mycelia in the wheat tissue is also primed for directing all of its resources into perithecia (inoculum) production. When high humidity is present, some of these resources will be spent on mycelium production. We are currently testing whether high humidity at harvest results in reduced inoculum the following growing season. We have investigated the physiology of these processes using Affymetrix expression arrays. We have identified a gene we believe may regulate the mycelium/no mycelium switch. We are pursuing the analysis of this gene. We investigated whether polyketides produced by F. graminearum are important for protecting the debris from invasion from other pathogens. We examined a set of mutants for each of the 15 polyketide synthase genes. We found some small effects of protection for 3 of the 15 polyketides. However, there was a much larger effect on the ability of these mutants to colonize the stems. Previous work showed that several PKS genes were expressed during stem colonization. The mutants of 2 of these no longer colonize stems, although they cause head blight disease. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have determined that relative humidity during harvest is likely to have a strong impact on ascospore production for the following year. We have also determined that secondary metabolites other than DON may affect the ability of the fungus to colonize wheat stems, resulting in inoculum the following year. Impact: The data on conditions for perithecium production vs. relative humidity at harvest is aimed at developing a component of forecasting systems which would take into consideration the amount of inoculum likely to be present. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: We have identified conditions which may greatly affect inoculum development in the fungus in the following season. In addition, if shown to hold in the field, a link between harvest conditions and inoculum development the following year would be readily incorporated into a forecasting system. (Form FPR07) 3

337 PI: Trail, Frances USDA-ARS Agreement #: Identification of a gene which regulates the switch between mycelia vs. perithecium production could be a target for genetically engineered resistance. Similarly, a gene which is important to colonization of stem tissue would also provide a target for reduction of inoculum production. Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Presentations (from 2007-present) Understanding the impact of amylase production on the life cycle of Fusarium graminearum. American Oil and Analytic Chemists International. Meeting Bozeman, MT, June Form and function in sexual development of Fusarium graminearum. Program for the Biology of Filamentous Fungi. 17 th annual symposium. Texas A and M, April Spore cannons and chemical warfare: The life cycle of a wheat pathogen. Presented to the Horticulture/ Crop and Soil Sciences joint departmental seminar series MSU. February Spore cannons and chemical warfare: The life cycle of Fusarium graminearum on wheat. Presented to the Department of Plant Biology at MSU. January Life cycle and survival of Fusarium graminearum- a revision. Presented to the Fusarium Head Blight Forum. Kansas City, Mo. December Integration of polyketides into the life cycle of Fusarium graminearum. 40 th Annual Meeting of the Society for Invertebrate Pathology, August, Quebec City, Quebec Identification of genes involved in sexual development in Gibberella zeae by expression analysis. Mycological Society of America Meeting, Baton Rouge, LA. Invited talk Weaving a life story: Studies on the disease cycle of Fusarium graminearum on wheat. Robert Gilmer Memorial Seminar. Plant Pathology Department, Cornell University Perithecium development and ascus function in Gibberella zeae. Invited talk. Fungal Genetics Conference, Asilomar, CA Related presentation: Spore cannons: Elucidating the function of asci. Gordon Conference on Cellular and Molecular Mycology. Plymouth New Hampshire, July Related papers (from 2007 to present) all of these papers were generated from preliminary studies that were funded by USWBSI that were used to obtain subsequent federal funding from other programs: Hallen, H., and F. Trail The L-type calcium ion channel, Cch1, affects ascospore discharge and mycelial growth in the filamentous fungus Gibberella zeae (anamorph Fusarium graminearum). Eukaryotic Cell. 7: (Form FPR07) 4

338 PI: Trail, Frances USDA-ARS Agreement #: C Cuomo, U Güldener, JR Xu, F Trail, BG Turgeon, A Di Pietro, JD Walton, L-J Ma, S Baker, M Rep, G Adam, J Antoniw, T Baldwin, S Calvo, Y-L Chang, D DeCaprio, LR. Gale, S Gnerre, RS Goswami, K Hammond-Kosack, LJ Harris, K Hilburn, JC Kennell, S Kroken, JK Magnuson, G Mannhaupt, E Mauceli, H-W Mewes, R Mitterbauer, G Muehlbauer, M Münsterkötter, D Nelson, K O'Donnell, T Ouellet, W Qi, H Quesneville, MI Roncero, K-Y Seong, IV Tetko, M Urban, C Waalwijk, TJ Ward, J Yao, BW Birren, HC Kistler The Fusarium graminearum genome reveals a link between localized polymorphism and pathogen specialization. Science 317: Hallen, H., Huebner, M., Shiu, S.-H., Guldener, U., and F. Trail Gene expression shifts during perithecium development in Gibberella zeae (anamorph Fusarium graminearum), with particular emphasis on ion transport proteins. Fungal Genetics and Biology 44: Trail, F Fungal cannons: Explosive spore discharge in the Ascomycota. FEMS Microbiology Letters 276: (Form FPR07) 5

339 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: Nilgun Tumer Institution: Rutgers, The State University of New Jersey Address: Biotech Center - Cook College 59 Dudley Rd. New Brunswick, NJ tumer@aesop.rutgers.edu Phone: x215 Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 43,190 Modification of the Ribosomal Target to Enhance Resistance to Trichothecene Mycotoxins. USWBSI Individual Project(s) USWBSI Research Area * GET ARS Adjusted Award Project Title Amount Modification of the Ribosomal Target to Enhance Resistance to $43,190 Trichothecene Mycotoxins. Total Award Amount $ 43,190 Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries

340 PI: Tumer, Nilgun USDA-ARS Agreement #: Project 1: Modification of the Ribosomal Target to Enhance Resistance to Trichothecene Mycotoxins. 1. What major problem or issue is being resolved and how are you resolving it? Our main goals in the project funded during the FY07 period was to develop DON resistant forms of the wheat L3 genes and to generate transgenic wheat plants with these genes to determine if overexpression of the mutant forms of the wheat L3 genes will confer resistance to FHB in wheat. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment: We have successfully demonstrated that expression of a truncated form of the yeast L3 gene (L3Δ), which corresponds to the first 99 amino acids of the yeast ribosomal protein L3, in transgenic wheat plants leads to improved resistance to Fusarium Head Blight (FHB) and a reduction in DON levels. During FY07 we have isolated genes encoding the wheat ribosomal L3 proteins. We have engineered the same mutations into the wheat L3 genes that previously conferred resistance to FHB in the yeast L3 genes. We prepared constructs with the DONresistant point-mutant forms of the wheat RPL3A1 gene and a truncated wheat RPL3A1 gene (L3Δ) downstream of the maize Ubiquitin1 promoter and first intron. Each coding region included V5 and 6xHis epitope tags so that the new proteins can be distinguished from native wheat L3. These constructs were provided to Dr. Ann Blechl for transformation into wheat. Dr. Blechl has produced several new transgenic wheat lines with the wheat L3Δ and the full-length wheat RPL3A1 mutants, W258K and W258C. We plan to identify homozygous lines and characterize expression of the wheat L3 genes. These lines will be evaluated in the greenhouse for resistance to FHB during FY08 and DON levels will be analyzed. Impact: We have demonstrated FHB resistance in transgenic wheat plants for the first time by expressing a modified form of the yeast ribosomal protein L3 gene. Our results also provided the first evidence that resistance to DON correlates with resistance to FHB and results in reduced accumulation of DON in transgenic wheat plants. We have now developed constructs containing the mutated forms of the wheat L3 genes and produced transgenic wheat plants with these genes. If transgenic wheat lines containing the modified wheat L3 genes show resistance to FHB and DON accumulation, these transgene loci will provide sources of novel resistance to FHB. These genes can be transformed into other wheat cultivars, such as the Hard Winter Wheat and Durum, as well as into barley plants. We anticipate that the transgenic wheat lines overexpressing the DON resistant forms of the wheat L3 genes may be more acceptable to the public. (Form FPR07) 2

341 PI: Tumer, Nilgun USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?: Developing effective FHB resistance through transgenics is one of the strategies that can be used to reduce the impact of FHB in wheat. Our work has demonstrated the feasibility of this approach. We have produced transgenic wheat lines that showed resistance to FHB and reduced DON accumulation in the greenhouse tests. During FY08, these lines were grown in a transgenic nursery established by the University of Minnesota and were included in the tours of this nursery that were open to the public. The transgenic lines are being evaluated in a FY08 field test at the University of Minnesota by Dr. Ruth Dill-Macky. If these lines show resistance in the field, they could be used as breeding material for introduction of the FHB resistance into elite wheat cultivars or breeding lines and can be combined with other sources of resistance to provide growers new wheat cultivars with effective FHB resistance. (Form FPR07) 3

342 PI: Tumer, Nilgun USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Publications J. E. McLaughlin, A. Bin Umer, J. Schifano, A. Tortora, S. McCormick, N. E. Tumer Genome wide analysis of trichothecene mycotoxin mechanisms of action in Saccharomyces cerevisiae. In preparation. Di, R., and N. E. Tumer. The N-terminal 99 amino acids of yeast ribosomal protein L3 reduce the toxicity of pokeweed antiviral protein in Saccharomyces cerevisiae. In preparation. Di, R., A. Blechl, R. Dill-Macky, A. Tortora, and N. E. Tumer Expression of the N- terminal 99 amino acids of yeast ribosomal protein L3 in transgenic wheat confers resistance to Fusarium head blight. Mol. Plant Breeding 5:283. Oral presentations in conferences N. E. Tumer Resistance to FHB by expression of modified L3 genes in wheat. National Fusarium Head Blight Forum Dec , Kansas City, MO Di, R., A. Blechl, R. Dill-Macky, A. Tortora, and N. E. Tumer Expression of the N- terminal 99 amino acids of yeast ribosomal protein L3 in transgenic wheat confers resistance to Fusarium head blight. The 2 nd International Conference of Plant Molecular Breeding, Hainan, China. (Form FPR07) 4

343 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: David Van Sanford Institution: University of Kentucky Address: Department of Agronomy 327 Plant Science Bldg. Lexington, KY Phone: ext Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 202,490 U.S. Wheat & Barley Scab Initiative Networking and Facilitation Office, and Website. USWBSI Individual Project(s) USWBSI Research Area * EC/HQ ARS Adjusted Award Project Title Amount U.S. Wheat & Barley Scab Initiative Networking and Facilitation $202,490 Office, and Website. Total Award Amount $ 202, _ Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)

344 PI: Van Sanford, David USDA-ARS Agreement #: Project 1: U.S. Wheat & Barley Scab Initiative Networking and Facilitation Office, and Website. 1. What major problem or issue is being resolved and how are you resolving it? Scab affects the industries and people involved in virtually every stage of the production, processing, and distribution systems of five market classes of wheat and barley across the U.S. The Networking and Facilitation Office (NFO) has been in existence since The goal of the NFO is to minimize the barriers to the success of the U.S. Wheat & Barley Scab Initiative arising from the administrative burdens and communication challenges. In FY07, there were 85 researchers working in 27 institutions (including ARS and CIMMYT) across 25 states and Mexico. The NFO is addressing this problem by 1) helping scientists and all interested parties, who in the past may have problems communicating with each other, to learn from each other by facilitating communications and communication system development; 2) identifying and implementing an internet-based communication and collaboration mechanisms; and 3) acting as a center of accountability and a rapid clearing house of scab-related information. The NFO provides administrative support to the various committees, as well as being responsible for the organization of the Initiative s annual scientific conference. Finally, the NFO facilitates the annual resolution of a comprehensive national research plan and budget, and represents the Initiative s only fixed asset. 2. List the most important accomplishment and its impact (how is it being used?). Complete all three sections (repeat sections for each major accomplishment): Accomplishment (1): During FY07, the USWBSI began implementation of its newly developed Action Plan starting with the creation of five coordinated Projects: Commodity-based Coordinated Projects: o Barley Coordinated Project o Hard Winter Wheat Coordinated Variety Development and Host Resistant Coordinated Projects o Spring Wheat Parents (included Durum projects) o Northern Winter Wheat o Southern Winter Wheat Impact (1): By coordinating research efforts within these specific categories, the expectation is that it will lead to greater collaboration and communication, improved focusing of research efforts and more efficient use of resources. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (1) Several of these coordinated projects include their stakeholders (growers, producers, endusers) in the development of the research plan for their commodity or region. Researchers have more involvement in determining the research priorities for their coordinated project. (Form FPR07) 2

345 PI: Van Sanford, David USDA-ARS Agreement #: Accomplishment (2): One of the NFO s most important accomplishments was the successful facilitation of the development of the USWBSI s FY08 Research Plan and Budget, which was approved by USDA-ARS in February. See attached table (1), for a summary of meetings, conference calls, etc. facilitated by the NFO that lead to the FY08 Research Plan and Budget. Impact (2): One-hundred and twenty-four research projects across five research areas, and the USWBSI Headquarters, received funding in the amount of $4,956,797 to continue the fight against FHB. As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before?:(2) There are 83 scientists in 25 states and Mexico continuing to work on developing as quickly as possible effective control measures that minimize the threat of Fusarium Head Blight (Scab) to the producers, processors, and consumers of wheat and barley. Accomplishment (3): In December of 2007, the NFO working with the Forum Organizing Committee facilitated the USWBSI s annual scientific Forum, which was held in Kansas City, Missouri. Over 200 crop scientists (including research associates, research technicians and graduate students), growers, and industry representatives from across the nation and around the world, as well as area leaders of the U.S. wheat and barley industry, attended the 2007 National FHB Forum. Research results and findings were presented to the forum participants in the form of invited talks (22), and poster (102) presentations accompanied by abstracts or papers published in the Forum proceedings. One afternoon was devoted to group discussions designed around the USWBSI s five research areas and three commodity-based coordinated projects. The focus of the group discussions was the implementation of the USWBSI s Action Plan and communicating results in the real world. There were also mini breakout sessions covering the following topics: Tebuconazole EPA Emergency Exemption: Recent Developments and 2008 Application Positioning; Genotyping Centers Breeders met with scientists from the genotyping labs. FHB Disease Forecasting Models; and Technicians and graduate students: an informal exchange of ideas and protocols for FHB research. Impact (3): The annual Forum is the primary mechanism whereby USWBSI scientists and stakeholders come together to discuss both the progress being made, as well as problems that interfere, with the successful combat against this devastating disease. (Form FPR07) 3

346 PI: Van Sanford, David USDA-ARS Agreement #: As a result of that accomplishment, what does your particular clientele, the scientific community, and agriculture as a whole have now that they didn t have before? (3) The clientele has a greater understanding of what research is being conducted under the USWBSI; as well what progress is being made in the following areas (titles of Forum poster sessions): Host Plant Resistance and Variety Development Gene Discovery and Engineering Resistance Pathogen Biology and Genetics Food Safety, Toxicology and Utilization of Mycotoxin-contaminated Grain FHB Management Table 1. Summary of Meetings, Conference Calls and Workshops facilitated by the Networking & Facilitation Office (NFO) during FY07. # of Meetings/ Committee Conference Call (CC) Description/Purpose of Meeting Steering Committee (SC) 2 Meetings : Semi-Annual Meeting : Semi-Annual Meeting Review FY08 Research Plan and Executive Committee (EC) 4 Meetings Budget Meetings: Follow-up to SC s meeting; met with review panels to discuss their recommendations; follow-up of Annual Forum and USWBSI s recommendation for FY08 Research Plan and Budget. USWBSI SC-EC Sub Committees Coordinated Project Committees (CPC) /Review Panels (RP) RA Workshops/ Planning Meetings 2 CC 1 Meeting 7 CC CPCs 8 CC RP 11 CC 4 Workshops/ Meetings 7 CC Conference Calls: Discuss USWBSI Business Annul Research Plan and Budget; setting of Working Caps; agenda for semi-annual spring SC meeting; and program for Annual Forum Communication sub-committee Meeting, Fargo, ND Communications sub-committee confernce call Purpose of meetings: Enhancing the USWBSI s communication activities EC sub-committee interview with Co-Chair candidates 2-08 PBG sub-committee - discussed future direction of PBG research area. Coordinated Project Committees (CPC) Discuss final FY08 coordinated project pre-proposal. Review Panels - Discuss FY08 Funding Recommendation 6-07 Barley Coordinated Project Planning Meeting, St. Paul, MN 6-07 DON Labs discuss protocols for sampling for DON VDHR Northern Winter Wheat Coordinated Project Conference Call 7-07 VDHR-Spring Wheat Coordinated Project Conference Call 8-07 FHB Management Biocontrol Planning Meeting, San Diego, CA 3-08 VDHR Soft Winter Wheat Coordinated Project Planning Meeting, Wooster, OH 3-08 PBG RAC conducted three separate conference calls with MGMT, GDER and VDHR research scientists/committee members FSTU discuss coordination of DON testing labs VDHR-Spring Wheat Coordinated Project Plannning Meeting, Fargo, ND (Form FPR07) 4

347 PI: Van Sanford, David USDA-ARS Agreement #: Include below a list of the publications, presentations, peer-reviewed articles, and non-peer reviewed articles written about your work that resulted from all of the projects included in the grant. Please reference each item using an accepted journal format. If you need more space, continue the list on the next page. Forum Proceedings: Canty, S., A. Clark, D. Ellis and D. Van Sanford (Eds.) Proceedings from the 2007 National Fusarium Head Blight Forum, Kansas City, MO; December 2-4. University of Kentucky, Lexington, KY. 252pp. Proceedings are available in PDF format on the USWBSI s Website ( Invited Talks: Van Sanford, D. A The USWBSI: Status Report Wheat Quality Council Annual Meeting. Kansas City, MO. February 19-21, Newsletters: Lilleboe, D., S. Canty, and D. Van Sanford. Spring Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 8, Issue 1. Lilleboe, D., S. Canty, and D. Van Sanford. Fall Fusarium Focus. U.S. Wheat & Barley Scab Initiative. Volume 7, Issue 2. Press Releases: News Release: Scab Incidence Varies Across Midwestern States News Release: Early Southern Scab Reports Mixed News Release: USWBSI Announces new Co-Chair News Release: Scab Disease Generally Low across U.S. in Press Release: USWBSI Mourns Loss of Co-Chair Tom Anderson. All Press Releases are available in PDF format on the USWBSI s Website ( (Form FPR07) 5

348 USDA-ARS/ U.S. Wheat and Barley Scab Initiative (approx. May 07 April 08) July 15, 2008 Cover Page PI: David Van Sanford Institution: University of Kentucky Address: Department of Agronomy 327 Plant Science Bldg. Lexington, KY Phone: ext Fax: Fiscal Year: 2007 USDA-ARS Agreement ID: USDA-ARS Agreement Title: FY07 ARS Award Amount: $ 50,598 Accelerating the Development of FHB-Resistant Soft Red Winter Wheat Varieties. USWBSI Individual Project(s) USWBSI Research Area * IIR ARS Adjusted Award Project Title Amount Accelerating the Development of FHB-Resistant Soft Red Winter $50,598 Wheat Varieties. Total Award Amount $ 50, Principal Investigator Date * CBCC Chemical, Biological & Cultural Control EEDF Etiology, Epidemiology & Disease Forecasting FSTU Food Safety, Toxicology, & Utilization of Mycotoxin-contaminated Grain GET Genetic Engineering & Transformation HGR Host Genetics Resources HGG Host Genetics & Genomics IIR Integrated/Interdisciplinary Research PGG Pathogen Genetics & Genomics VDUN Variety Development & Uniform Nurseries (Form FPR07)