2006 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? Diseases and pests limit barley productivity in the United States, reducing farm income and the availability of high quality barley for food and feed industries. Fusarium head blight (FHB) infestation has been the most challenging disease problem in barley for more than a decade, with an economic impact in the billions of dollars. In the major upper Midwest production area, farmers have suffered both yield losses due to FHB and quality losses due to accumulation of the mycotoxin, deoxynivalenol, which makes barley unsuitable for malting and food use. Few resistance sources have been found in a worldwide search. Resistance in these genotypes is quantitative and even the best lines have unacceptable toxin levels when FHB severity is high. Other diseases and pests cause variable yield and quality losses each year. Russian wheat aphid (RWA) has become an increasing problem causing reduced yields in the western United States. The goal of this research project is to increase barley resistance to these diseases and pests, by providing new genes and linked markers for breeding resistance to FHB and RWA, and better understanding of the genes involved in resistance. Both traditional and transgenic approaches will be used. Related research into the involvement of ethylene in green plant regeneration will improve tissue culture methods for more efficient genotype-independent transformation with candidate resistance genes so researchers can insert new genes into their cultivar of choice rather than being restricted to unadapted model cultivars. Characterization of genes in the ethylene pathway may provide mechanisms for manipulating ethylene regulation in response to pathogen attack. This research is relevant to the National Program 302 Action Plan Component 2, Biological Processes that Improve Crop Productivity and Quality, Problem Statement 2B: Understanding Plant Interactions with Their Environment, and National Program 301 Action Plan Problem Area 2A: Genome Characterization. This plan includes a research objective to develop an efficient, genotype-independent transformation system in barley, which is needed to evaluate the effects of newly isolated disease resistance genes. Development of the new transformation methods is relevant to Component 3. Plant Biotechnology Risk Assessment, Problem Statement 3A: Improving and Assessing Genetic Engineering Technology. 2.List by year the currently approved milestones (indicators of research progress) 2006 Determine if there is a relationship between ethylene production in tissue culture and green plant regeneration rates. Determine if manipulating ethylene in culture can alter green plant regeneration in a high regenerating cultivar and a low regenerating cultivar. Conduct disease testing of backcross-derived lines expressing the transgenes chitinase and thaumatin-like protein. 2007 FHB: Complete molecular testing of BC2F2, select lines and increase; RWA: Complete map saturation on STARS-9301B Ethylene: Complete 1st experiments on detailed ethylene control; Transformation: Complete analysis of T1 plants; FHB field trials of previously identified homozygous lines 2008 FHB: Field trials of selected lines RWA: Complete tests of new STARS-9577B population for RWA reaction Ethylene: Complete 2nd experiments on detailed ethylene control Transformation: Select additional homozygous T2 lines and conduct field trials for FHB reaction; analyzed new transgenics 2009 FHB: Field trials of selected lines RWA: Complete map saturation on STARS-9577B Ethylene: Complete isolation of genes from at least 2 ethylene gene families Transformation: Complete backcrosses on the 1st 40 events; begin testing for other fungal diseases; FHB field trials 2010 FHB: Complete fine-mapping the selected QTLs; RWA: Complete analysis of the testcross population Ethylene: Complete mapping ethylene genes and comparison to QTLS; Transformation: FHB field trials 2011 FHB: Develop transgenic lines expressing candidate resistance genes; Ethylene: Complete expression assays in tissue culture; Transformation: Complete backcrosses on the remaining events; FHB field trials 4a.List the single most significant research accomplishment during FY 2006. Role of ethylene in green plant regeneration from tissue culture: Manipulation of ethylene in barley tissue cultures resulted in increased green plant regeneration, even from Golden Promise, a cultivar used for its high regeneration capabilities. Barley tissue culture is used for many procedures, especially for inserting new genes, and has been limited to model cultivars because of poor regeneration from most commercially adapted cultivars. Experiments with Golden Promise and Morex cultures increased ethylene production by incorporating the precursor ACC or inhibited ethylene action by the addition of silver nitrate, with treatments at specific stages of culture. Increased ethylene reduced regeneration in Golden Promise but at certain stages increased regeneration in Morex, while silver nitrate treatment during culture maintenance stages increased regeneration from both cultivars. The timing of ethylene action appears to be critical for maximum regeneration and specific manipulations of ethylene were identified that can be used to increase plant regeneration from tissue cultures of barley and other species, to allow scientists to select the best cultivars for their experiments. This accomplishment is relevant to the National Program 302 Action Plan Component 3. Plant Biotechnology Risk Assessment, Problem Statement 3A: Improving and Assessing Genetic Engineering Technology. 4b.List other significant research accomplishment(s), if any. None. 4c.List significant activities that support special target populations. None. 4d.Progress report. See subordinate projects. 5.Describe the major accomplishments to date and their predicted or actual impact. This is a new project begun in March 2006. This research will identify locations and effects of individual Fusarium head blight and RWA resistance genes, plus linked molecular markers for marker-assisted selection. Research on ethylene will give barley researchers the ability to culture and transform most genotypes, rather than just the few that can be transformed now, so scientists can select the best targets for expression of their genes of interest and add resistance to adapted cultivars that are susceptible to diseases such as FHB. Testing of transgenic lines will give a better understanding of factors influencing transgene expression, and knowledge of which transgenes improve resistance to diseases like FHB. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Provided transgenic barley seed to North Dakota State University, ARS-Aberdeen, the University of Idaho, for disease testing and backcrossing; provided transgenic leaf tissue to the University of Wisconsin for testing transgene toxin specificity; provided advice and information on barley tissue culture and transformation to scientists in Egypt, Bulgaria, Argentina and the Czech Republic; provided information on barley FHB resistance QTLs and linked markers to scientists in the Czech Republic. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). See the final report for 5442-21000-022-00D.