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? Barley and oat represent important commodities for feed, food, and malt use in the United States. The viability of U.S. industries that use these crops depends on continuing improvements in the efficiency with which they can be grown and used, e.g., increased yield and decreased cost of production. In addition, the worth of these commodities must be increased. The value of barley and oat could be enhanced by incorporating characteristics contributing to more environmentally friendly production systems and by increasing the health-promoting properties of the final products. The need for such improvements is particularly important in light of the continuing erosion of U.S. markets as a result of competitive imports. The U.S. cannot afford to lose markets for barley and oat as they represent important components in environmentally sound crop rotations. Also, in many regions of the northern U.S. there are few alternative crops which are adapted to limitations imposed by such factors as short growing seasons and low water availability. A great diversity of barley and oat germplasm is available and forms the genetic basis for our efforts to improve productivity, nutritional and other end-use qualities, disease and insect resistance, and to reduce the environmental impacts associated with the growth of barley and oat. Germplasm resources include accessions from the National Small Grains Collection (NSGC), specialized genetic stocks derived from mutation and transformation experiments, and specialized and elite breeding lines and cultivars developed at Aberdeen and at other locations. This project has three specific goals:. 1)Develop, characterize, and deploy improved barley and oat cultivars. Oat cultivar development will focus on hulled and hulless oat varieties and germplasm adapted to oat production regions throughout the U.S. Barley cultivar development will focus on winter malting types and spring feed and malting types specifically adapted for the intermountain west region. . 2)Generate improved barley and oat germplasm by moving traits from accessions in the NSGC and other sources into improved plant types, focusing on barley for food, barley and oat for feed (particularly aquaculture feeds), and improved disease and insect resistance. . 3)Develop, evaluate, and apply technologies such as genetic transformation and marker assisted selection and devise methods for their routine application to germplasm enhancement and variety development. This research is directed primarily towards the goals of NP 301, Plant Genetic Resources, Genomics and Genetic Improvement; Component II-Genome Characterization and Genetic Improvement; Problem Areas IIa-Genome Characterization, and IIb-Genetic Improvement. The development of enhanced oat and barley cultivars and germplasm--and of the new tools required to improve the developmental process--will lead to increased productivity, utility, profitability, and environmental sustainability of these two crops. Greater utility for feed and food uses will result from improving end-use quality, and by developing novel special-use barley and oat cultivars for food and feed uses, thus increasing marketing opportunities. Environmental sustainability of commercial agriculture will be enhanced by producing cultivars which are pest-resistant, adapted to dryland conditions, and which produce grain with enhanced phosphorous availability. 2.List by year the currently approved milestones (indicators of research progress) Year 1 (FY 2003) Multilocation trials established. Barley enhanced for beta-glucan and protein content available for nutritional studies. EST-SSR survey for oat completed. PCR-based markers for DP selection in barley developed. Year 2 (FY 2004) 30 RWA resistant lines released. LP oat mutants identified. Enhanced oil oat germplasm identified and available for fish diet tests. Six-rowed spring barley germplasm with enhanced diastatic power is identified and is available for further testing. Agronomic characterization and quality analysis completed for transgenic wheat and barley. Year 3 (FY 2005) Stripe rust resistant cultivar and RWA-resistant cultivar released. Agronomically enhanced LP barley lines available for fish diet tests. Oat map consensus markers identified and co-mapped. Year 4 (FY 2006) Superior winter malting barley lines available for further evaluation. LP oat germplasm available for evaluation. AcTPase (-) lines with antifungal transgenes identified. DP barley isogenics developed via marker- assisted backcrossing. Year 5 (FY 2007) Improved six-row malting cultivar candidate available for evaluation in intermountain region. New sources of partial resistance to oat crown rust identified and characterized. Recombinant populations for transposon tagging characterized and available for distribution. Oat consensus map developed. 4a.List the single most significant research accomplishment during FY 2006. This accomplishment aligns with the problem statement "Germplasm Enhancement/Release of Improved Genetic Resources and Varieties" in the NP 301 Action Plan Release of 'Herald' low-phytate barley 'Herald,' a feed barley with significantly reduced levels of phytate, has been released to the public. Phytate is a phosphorus-containing compound that is indigestible by non-ruminant animals, and which is excreted in manure where it can become a significant source of phosphorus pollution in surface and ground water. The digestibility of phosphorus in low-phytate barley is enhanced, and animals fed low-phytate barley require less phosphorus supplementation and consequently excrete less phosphorus. The development of Herald was conducted at the National Small Grains Germplasm Research Facility in Aberdeen, Idaho, and was made possible by the isolation of low-phytate mutant barley lines which were subsequently used as parents in a hybridization program designed to combine the low-phytate trait with good agronomic performance. Widespread adoption and use of Herald would have a positive environmental impact by reducing phosphorus generated by feeding animals, and contribute to producer profitability by reducing or eliminating the need for phosphorus supplements. 4b.List other significant research accomplishment(s), if any. Release of 'Sublette' malting barley 'Sublette' a two-rowed spring malting barley was approved for plant-scale evaluation by the American Malting Barley Association and released in 2005. It is currently being grown on 250 acres in southeast Idaho for evaluation in plant-scale trials by several major brewing companies. This variety is similar to the industry standard 'Harrington' with improvements in several important malting characteristics as well as improved yield potential. This variety could potentially improve producer profitability due to its increased yield potential and improved straw strength which could require less use of costly growth inhibitors to prevent lodging. This accomplishment aligns with the problem statement "Germplasm Enhancement/Release of Improved Genetic Resources and Varieties" in the NP 301 Action Plan. Improved assessment of crown rust resistance in oat An improved method of measuring crown rust resistance in oat was developed. Crown rust is the most important disease of oat and more efficient ways to breed for resistance to the disease are needed. We developed a new method to assess crown rust resistance at the Small Grains and Potato Germplasm Research Unit in Aberdeen, ID by measuring the amount of pathogen DNA within oat tissues. We anticipate that the new method will enable us to more precisely identify and map QTL for crown rust resistance. This accomplishment aligns with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. Characterization and comparison of transgenic wheat and barley lines for agronomic performance Analyses of the agronomic performance of transgenic wheat lines and comparisons to previous observations of transgenic barley lines were completed. This study addresses the need for objective data concerning the extent to which agronomic performance is affected by transformation procedures, data that is necessary for the development of efficient breeding schemes for the use of transgenic parents. The design and coordination of this work was accomplished at the ARS National Small Grains Germplasm Research Facility, development of the lines tested and the conception of these studies was done at the ARS Western Regional Research Center, and field and laboratory analyses were conducted at Aberdeen and by University of California collaborators at Davis and El Centro, CA. The results of this research make a substantial contribution to a small but critical body of literature concerning the performance and utility of transgenic cereals, and will be used by geneticist and breeders as they design strategies for the efficient use of transgenic cereal germplasm. This accomplishment aligns with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. Agronomic and quality characterizations of low-phytate barley germplasm Analyses of agronomic performance, grain phosphorus characteristics, and malting quality of barley lines as affected by four independent low-phytate mutations were completed. These studies addressed the need for objective data concerning the effects of these mutations on agronomic and quality characteristics in various production environments. Scientists at the ARS National Small Grains Germplasm Research Facility in Aberdeen, ID developed all materials used for these tests and produced the agronomic and phosphorus data and malting quality characteristics were assessed at the ARS Cereal Crops Research Unit in Madison, WI. This research resulted in thorough characterization of these four mutations that verified their unique influences on agronomic performance and phosphorus characteristics clearly identified the mutations that combine both acceptable performance and the low-phytate trait. The results will guide geneticists and breeders in the appropriate selection and use of this germplasm for additional genetic research and the production of commercially useful low-phytate barley germplasm. This accomplishment aligns with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. Development and distribution of recombinant populations for transposon tagging A set of specialized lines that will enable transposon tagging in barley has been assembled, characterized, and made ready for distribution to other scientists. In 2006, two objectives critcal to the implementation of these resources were achieved: development and use of analytical procedures for verifying the expression of Ac transpose, and the production and verification of the identity of various lines containing different Ds inserts. This accomplishments addresses the need for additional tools for genomic research in barley. The specific work listed above was conducted at the ARS National Small Grains Germplasm Research Facility in Aberdeen, ID, but is part of a larger collaborative effort, funded by the NSF, involving researchers at the University of California that were responsible for producing the original germplasm used in this research, and researchers at Oregon State University who mapped the location of insertions. This tool will enable the production of mutants with altered phenotypes, and the identification, characterization, and cloning of genes responsible for such alterations. To date, distribution of Ac transposase and Ds lines have been made to three researchers outside of this project. This accomplishment aligns with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. Identification of the cause of inviability in hybrids between Ds-containing lines and Oregon Wolfe Barley Dominant (OWBD) multiple marker stock The hypothesis that inviability in Ds/OWBD hybrids was caused by high levels of expression of the selectable marker gene bar was definitively demonstrated. Extensive phenotypic, ELISA, and quantitative RT-PCR analyses were completed and and compiled into a report which has been submitted for peer-reviewed publication. This study was conducted to identify the cause of an unexpected phenomenon--the inability to produce mapping populations between OWBD and Ds-containing lines--and addresses the need to thoroughly characterize transgenic barley germplasm as part of our duty to examine the consequences of new methods of effecting genetic modifications. This work was done at the ARS National Small Grains Germplasm Research Facility, utilizing materials and information generated as part of a collaborative effort involving researchers at the University of California and at Oregon State University. The results of this study suggest the need for, and enable further studies of, the genetic explanation of this phenomenon and the implications for the use of bar in the production of transgenic barley. This accomplishment aligns with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. 4c.List significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments to date and their predicted or actual impact. The following accomplishments align with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. Release of the Russian wheat aphid-resistant barley cultivar 'Burton.' This is the first RWA-resistant barley cultivar, and it will re-enable the production of barley without the use of insecticides in several areas of the intermountain region of the Western United States. Foundation seed of Burton, produced via collaboration with the University of Idaho Foundation Seed program, has been purchased by seed producers in Colorado and Nebraska for distribution to farmers in 2006. The low-phytate barley 'Herald' was released in 2006. Widespread adoption and use of Herald would have a positive environmental impact by reducing phosphorus generated by feeding animals, and contribute to producer profitability by reducing or eliminating the need for phosphorus supplements. 'Sublette' a two-rowed spring malting barley was approved for plant-scale evaluation by the American Malting Barley Association and released in 2005. This variety could potentially improve producer profitability due to its increased yield potential and improved straw strength which could require less use of costly growth inhibitors to prevent lodging. We developed hulless barley germplasm with high beta-glucan levels (8 to 9%), high protein (>15%) and germplasm with different compositions of amylose/amylopectin starch (2 to 35% amylose or 65 to 98% amylopectin). This hulless barley germplasm will be valuable in the food and feed industry. 'Charles,' a new winter malting barley, was released. This is the first winter barley to meet industry specifications for malt quality in pilot scale evaluations. It is being increased for evaluation in plant-scale by several major brewing industry companies. Multi-location barley evaluations in 13 environments in Idaho, Oregon, and Montana were established. These evaluations will be critical to identifying widely adapted germplasm for the region. The following accomplishments align with component III, Genetic Improvement of Crops, and the problem statement "Genetic Theory and Methods of Crop Improvement" in the NP301 Action Plan. A new means of assessing crown rust resistance in oat was developed by measuring the amount of pathogen DNA in oat tissues. The new technique could greatly enhance our ability to identify QTL for crown rust resistance and improve our ability to breed for crown rust resistance. A recombinant inbred (RI) oat mapping population was developed from a cross between 'Ogle' and 'TAMO-301.' This population, commonly referred to as "OT," has been used to identify and map several quantitative trait loci (QTL) controlling agronomic and quality traits in oat. Using OT, we collaborated with plant pathologists to identify disease resistance genes. OT will continue as an important resource for oat geneticists to characterize economically important genes and be useful in the development of an oat consensus map. Analyses of the agronomic performance of transgenic wheat lines and comparisons to previous observations of transgenic barley lines were completed. This study addresses the need for objective data concerning the extent to which agronomic performance is affected by transformation procedures, data that is necessary for the development of efficient breeding schemes for the use of transgenic parents. The design and coordination of this work was accomplished at the ARS National Small Grains Germplasm Research Facility, development of the lines tested and the conception of these studies was done at the ARS Western Regional Research Center, and field and laboratory analyses were conducted at Aberdeen and by University of California collaborators at Davis and El Centro, CA. The results of this research make a substantial contribution to a small but critical body of literature concerning the performance and utility of transgenic cereals, and will be used by geneticist and breeders as they design strategies for the efficient use of transgenic cereal germplasm. Analyses of agronomic performance, grain phosphorus characteristics, and malting quality of barley lines as affected by four independent low-phytate mutations were completed. These studies addressed the need for objective data concerning the effects of these mutations on agronomic and quality characteristics in various production environments. Scientists at the ARS National Small Grains Germplasm Research Facility in Aberdeen, ID developed all materials used for these tests and produced the agronomic and phosphorus data and malting quality characteristics were assessed at the ARS Cereal Crops Research Unit in Madison, WI. This research resulted in thorough characterization of these four mutations that verified their unique influences on agronomic performance and phosphorus characteristics, clearly identified the mutations that combine both acceptable performance and the low-phytate trait. The results will guide geneticists and breeders in the appropriate selection and use of this germplasm for additional genetic research and the production of commercially useful low-phytate barley germplasm. 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? Information on present and future barley cultivars was presented at the following grower meetings: US Grains Council International Malt Buyers Conference, Portland, OR University of Idaho Cereal Field Tour, Aberdeen, ID Caribou County Field Day-Soda Springs, ID Idaho Barley Commission Meeting, Boise ID Review Publications Bregitzer, P.P., Mornhinweg, D.W., Hammon, R., Stack, M., Baltensperger, D.D., Hein, G.L., O'Neill, M.K., Whitmore, J.C., Fiedler, D.J. 2005. Registration of burton barley. Crop Science. 45:1166-1167 Bregitzer, P.P., Raboy, V. 2006. Effects of four independent low-phytate mutations in barley (hordeum vulgare l.)on seed phosphorus characteristics and malting quality. Cereal Chemistry. Crop Sci. 46:1318-1322 (2006) Cooper, L.D., Marquez-Cedillo, L., Singh, J., Zhang, S., Carollo, V.L., Edwards, V.M., Johnson, K., Klienhofs, A., Rangel, S., Sturbaum, A.K. 2004. Mapping ds insertions in barley using a sequence-based approach. Molecular Genetics and Genomics. 271: 181-193. Mornhinweg, D.W., Obert, D.E., Wesenberg, D., Erickson, C.A., Porter, D.R. 2006. Registration of seven winter feed barley germplasms lines resistant to Russian wheat aphid. Crop Science. 46:1826-1827. Jackson, E.W., Avant, J.B., Overturf, K.E., Bonman, J.M. 2006. A quantitative assay of puccinia coronata f.sp. avenae dna in avena sativa.. Plant Disease. 90:629-636. DOI:10.1094/PD-90-0629 Hang, A., Bockelman, H.E., Burton, C.S. 2005. Cytological and seed morphological investigation of 250 accessions from the w.j. sando collection.. Agronomy Society of America, Crop Science Society of America, Soil Science Society of America Meeting, November 6-10, 2005, Salt Lake City, Utah. 2005 CDROM Bregitzer, P.P., Fiedler, D.J., Blechl, A.E., Lin, J.W., Sebesta, P.G., Fernandez De Soto, J., Chicaiza, O., Dubcovsky, J. 2006. Changes in high molecular weight glutenin subunit composition can be genetically engineered without impacting wheat agronomic performance. Crop Science. 46:1553-1563 (2006) Erickson, C.A., Obert, D.E., Burrup, D.E., Whitmore, J.C., Wesenberg, D.M. 2006. Registration of creel barley. Crop Science. 46:1812-1813. Obert, D.E., Wesenberg, D.M., Burrup, D.E., Jones, B.E., Erickson, C.A. 2006. Registration of 'charles' winter malting barley. Crop Sci 46:468-469 Obert, D.E., Wesenberg, D.M., Burrup, D.E., Erickson, C.A., Whitmore, J.C., Jones, B.E. 2006. Registration of `sublette¿ spring malting barley. Crop Sci 46:989-991