Progress 06/03/10 to 06/02/15

Outputs
Progress Report Objectives (from AD-416): Identify milling, baking, and pasta processing traits of experimental and advanced lines of hard spring and durum wheat that impact end-use quality and the overall development and release of new commercial cultivars of wheat. Identify specific biochemical components in wheat that are associated with end-use baking quality traits. More specifically, to develop rapid methods to characterize proteins, which are initially important for predicting end-use quality; to determine the variation in free asparagine content in wheat, which is an important precursor for the formation of carcinogenic acrylamide during bread baking and toasting; and to identify the efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat. Optimize methodologies for the extraction and analysis of soluble oat fiber components; determine environmental variation in the quantity and quality of �-glucan as characterized in the first objective in newly developed high �-glucan cultivars and other oat cultivars; and determine relationships between oil concentration and �-glucan concentration in segregating crosses for these traits, as to how each affects groat breakage during dehulling. Approach (from AD-416): Approximately 4000 hard spring and 1000 durum wheat experimental lines will be submitted by Federal, State, and private breeders, and special interest groups on an annual basis and evaluated for end-use quality. Wheat will be obtained from different generations of development in specific breeding programs, which will include early-generation, preliminary, and advanced lines and include commercial cultivars as controls. Measureable end-use quality traits include test weight, size, seed weight, sprout damage, physical evidence of disease, and protein and ash content; milling performance as a function of flour and semolina yield; flour and semolina color, protein, and ash content; dough strength properties; bread baking quality; and pasta processing quality. The Fargo Genotyping Laboratory will identify quantitative trait loci regions associated with end-use quality, primarily kernel and dough strength traits on selected hard spring wheat samples. Research will be conducted on the rapid characterization of protein extracts from wheat and flour by testing two different protein extraction methods. Variation in free asparagine content in wheat lines will be measured. The efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat, specifically protein composition and amino acid profiles, will be determined as a function of dough strength characteristics. To develop optimized methodologies for the extraction of beta-glucan from oats, samples of oats will be obtained from replicated plots grown in diverse environments over several years. The newly developed methods for the analysis of soluble fiber quality will be used to determine how environment might affect the quality and concentration of beta-glucan, especially in newly developed high beta-glucan cultivars. Beta-glucan effects on milling quality of oats will also be tested. Because beta- glucan is in the cell walls of oats, it is thought to provide a strengthening effect, which reduces groat breakage during dehulling. However, high oil in oats also seems to prevent groat breakage, and currently breeders have been unable to separate the traits of high oil and high beta-glucan in oats. A collaborating genetics program has developed populations of recombinant inbred lines from oat crosses designed to be segregating for high oil and high beta-glucan. We will evaluate the phenotypes of these lines and test their behavior during dehulling to determine the relative roles of oil and beta-glucan in preventing groat breakage during oat milling. This is the final report for Project 3060-43440-011-00D, which has been replaced by Project 3060-43440-014-00D. Please refer to the latter project for future reports. The Hard Red Spring Wheat Quality Laboratory (WQL) has analyzed physical and biochemical quality traits of approximately 3000 samples of hard spring and durum wheat every year. Samples were submitted by private and public wheat breeders involved in wheat germplasm improvement and by scientists involved in gene mapping to identify quantitative trait loci associated with end-use quality traits. The WQL provided over 40 different tests related to wheat kernel characteristics, milling performance, and flour, semolina, dough, baking, and spaghetti processing quality. This program contributed to the release of three hard spring wheat cultivars from the University of Minnesota, four hard spring wheat cultivars from South Dakota State University, one mapping population from the University of Minnesota, and one winter durum wheat cultivar from Virginia Tech during the last five years. In cooperation with the Wheat Quality Council (WQC), approximately ten experimental lines of hard spring wheat that were grown at up to 5 locations have been evaluated along with check cultivars every year. The WQL tested each line for kernel, milling, flour, dough, and bread-baking quality traits, coordinated baking quality test results from independent public and private testing laboratories, and analyzed the data. Results were published, presented, and discussed at the annual meeting of the WQC. Interactions with the WQC serve as a means to obtain industry feedback on the milling and baking quality traits of advanced experimental lines of wheat being considered for release into commercial production. A 2013 variety survey showed that all of the top five HS wheat cultivars planted in North Dakota, Minnesota, and South Dakota were tested by the WQC program. Research regarding the effects of protein molecular weight distribution (MWD) on bread-making characteristics of hard spring wheat was continued. We improved the speed of size-exclusion high performance liquid chromatography (SE-HPLC) for analysis of protein MWD and identified protein fractions that had significant associations with quality characteristics. This information can contribute to rapid characterization of protein for quality evaluation of a large number of hard spring wheat samples. Flour samples of hard spring wheat genotypes grown in North Dakota and Minnesota were also analyzed by SE-HPLC and evaluated by UV spectroscopy for three years. The data obtained from this research is currently being evaluated to develop prediction models of quality traits and to assess effects of genotype and growing environment on protein MWD parameters and their associations with wheat quality traits using statistical and multivariate data analysis methods. Research also was performed to assess the suitability of SE-HPLC for quality evaluation of durum semolina using semolina samples that were milled from 13 durum genotypes grown at 7 locations over a three year period in North Dakota. Firmness of cooked pasta is an important quality characteristic for durum wheat, and this research identified protein fractions that had significant effects on firmness of cooked pasta. The SE-HPLC data is being further analyzed to assess possible effects of genotype and environment on protein SE-HPLC parameters and their associations with durum wheat quality characteristics. UV spectroscopy data was also collected and will be evaluated for durum wheat quality evaluation as part of the next project. Altogether, this information can contribute to more rapid quality evaluation of durum wheat breeding lines. The segregation of low asparagine wheat genotypes is important to reduce carcinogenic acrylamide content in baking products. We identified significant variation in free asparagine concentration for flour mill streams, hard spring wheat genotypes damaged by pre-harvest sprouting, and durum translocation lines. Free asparagine concentration was also analyzed for hard spring wheat genotypes collected from North Dakota and Minnesota for three years, and the data are being statistically analyzed to investigate the variation in free asparagine among hard spring wheat genotypes. The information obtained is important to millers in blending of flour mill streams and to breeders in selection of wheat genotypes with low asparagine concentration, which may contribute to reducing carcinogenic acrylamide in baked products. An investigation into the effect of sulfur fertilization on quality characteristics of hard spring wheat genotypes was initiated using six wheat genotypes that were grown at three different levels of sulfur fertilization at three locations in MN. The wheat samples are currently being analyzed for milling and baking quality, and the data will be statistically analyzed to assess the effect of sulfur fertilization on protein, free asparagine, and quality characteristics. Wheat kernel vitreousness is an important grading characteristic in the U.S., and therefore the effect of kernel vitreousness on milling and breadmaking quality in hard red spring wheat was investigated. Data analysis indicated that wheat quality characteristics are mainly associated with protein composition related to molecular weight distribution, and kernel vitreousness is a good supplementary trait to protein for evaluation of milling and breadmaking quality characteristics such as break flour yield and baking water absorption. The solvent retention capacity (SRC) test measures the ability of flour to retain a set of four solvents as related to intrinsic flour constituents. When the suitability of the SRC test for quality assessment of hard red spring wheat flour was examined, we identified a discrepancy for the association between SRC parameters, protein fractions analyzed by size-exclusion HPLC, and breadmaking characteristics that could help segregation of HRS wheat flour samples for breadmaking quality. Pasta fortification with non-traditional ingredients was investigated with the purpose of offering healthier alternatives to consumers. Spaghetti samples that were made from traditional and non-traditional formulations, including whole wheat flour and flaxseed flour, were analyzed for cooking quality and biochemical components. Among protein fractions, high levels of a protein fraction called glutenin polymers and low levels of the albumin and globulin fractions were identified to have association with low cooking loss and weight, and high firmness for the cooked non-traditional spaghetti. This research provides novel information on the influence of biochemical components on cooking quality of nontraditional pasta that may help formulate flour mixes for non- traditional pasta products. Intermediate wheatgrass (Thinopyrum intermedium, IWG), possesses desirable agronomic traits, e.g., disease resistance, relatively large seed, and grain yield. As a perennial relative of wheat, IWG has good potential for development into a perennial grain crop. To investigate the variability of end-use quality among IWG genotypes, we evaluated protein composition and mixing properties for whole grain flour samples of 60 IWG genotypes. This research provided quality trait data and their associations with protein composition parameters for IWG genotypes grown in MN. Overall, this information will benefit the evaluation and improvement of quality and improve breeding efficiency for IWG. Accomplishments 01 Development of improved wheat germplasm. Wheat producers, milling and baking industries, and overseas customers require high standards in the quality of wheat to meet their evolving needs. The Hard Red Spring and Durum Wheat Quality Laboratory contributed wheat end-use quality data that helped lead to the development of improved wheat germplasm and subsequent release of new cultivars of spring and durum wheat bred for commercial production. Such new releases provide value to both farmers and industry through improved productivity or end-use quality.

Impacts
(N/A)

Publications

• Johnson, C.R., Thavarajah, D., Thavarajah, P., Payne, S., Moore, J., Ohm, J.-B. 2015. Processing, cooking, and cooling affect prebiotic concentrations in lentil (Lens culinaris Medikus). Journal of Food Composition and Analysis. 38:106-111.
• Whitney, K., Ohm, J.-B., Simsek, S. 2014. Addition of glucose oxidase for the improvement of refrigerated dough quality. Cereal Chemistry. 91(6):548- 553.
• Baasandorj, T., Ohm, J.-B., Manthey, F., Simsek, S. 2015. Effect of kernel size and mill type on protein, milling yield, and baking quality of hard red spring wheat. Cereal Chemistry. 92(1):81-87.
• Collison, A., Yang, L., Dykes, L., Murray, S., Awika, J.M. 2015. Influence of genetic background on anthocyanin and copigment composition and behavior during thermoalkaline processing of maize. Journal of Agricultural and Food Chemistry. 63:5528-5538.
• Ibrahim, A., Rudd, J., Devkota, R., Baker, J., Sutton, R., Simoneaux, B., Opena, G., Herrington, R., Rooney, L., Dykes, L., Awika, J., Nelson, L.R., Fritz, A., Bowden, R.L., Graybosch, R.A., Jin, Y., Seabourn, B.W., Chen, X. , Kolmer, J.A., St Amand, P., Bai, G., Duncan, R. 2015. Registration of 'TAM 305' hard red winter Wheat. Journal of Plant Registrations. doi:10. 3198/jpr2014.08.0054crc.
• Cremer, J.E., Bean, S., Tilley, M., Ioerger, B.P., Ohm, J., Kaufman, R.C., Wilson, J.D., Innes, D., Gilding, E.K., Godwin, I.D. 2014. Grain sorghum proteomics: An integrated approach towards characterization of seed storage proteins in kafirin allelic variants. Journal of Agricultural and Food Chemistry. 62:9819-9831.
• Anderson, J.A., Wiersma, J.J., Linkert, G.L., Reynolds, S., Kolmer, J.A., Jin, Y., Dill-Macky, R., Hareland, G.A. 2015. Registration of 'Rollag' spring wheat. Journal of Plant Registrations. 9:201-207.
• Sivapragasam, N., Thavarajah, P., Ohm, J.-B., Thavarajah, D. 2014. Surface properties of semi-synthetic enteric coating films: Opportunities to develop bio-based enteric coating films for colon- targeted delivery. Bioactive Carbohydrates and Dietary Fibre. 4:139-143.
• Glover, K.D., Hall, R.G., Jin, Y., Osborne, L.E., Ingemansen, J.A., Turnipseed, E.B., Hareland, G.A. 2015. Registration of 'Advance' Hard Red Spring Wheat. Journal of Plant Registrations. 9:83-88.

Progress 10/01/13 to 09/30/14

Outputs
Progress Report Objectives (from AD-416): Identify milling, baking, and pasta processing traits of experimental and advanced lines of hard spring and durum wheat that impact end-use quality and the overall development and release of new commercial cultivars of wheat. Identify specific biochemical components in wheat that are associated with end-use baking quality traits. More specifically, to develop rapid methods to characterize proteins, which are initially important for predicting end-use quality; to determine the variation in free asparagine content in wheat, which is an important precursor for the formation of carcinogenic acrylamide during bread baking and toasting; and to identify the efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat. Optimize methodologies for the extraction and analysis of soluble oat fiber components; determine environmental variation in the quantity and quality of �-glucan as characterized in the first objective in newly developed high �-glucan cultivars and other oat cultivars; and determine relationships between oil concentration and �-glucan concentration in segregating crosses for these traits, as to how each affects groat breakage during dehulling. Approach (from AD-416): Approximately 4000 hard spring and 1000 durum wheat experimental lines will be submitted by Federal, State, and private breeders, and special interest groups on an annual basis and evaluated for end-use quality. Wheat will be obtained from different generations of development in specific breeding programs, which will include early-generation, preliminary, and advanced lines and include commercial cultivars as controls. Measureable end-use quality traits include test weight, size, seed weight, sprout damage, physical evidence of disease, and protein and ash content; milling performance as a function of flour and semolina yield; flour and semolina color, protein, and ash content; dough strength properties; bread baking quality; and pasta processing quality. The Fargo Genotyping Laboratory will identify quantitative trait loci regions associated with end-use quality, primarily kernel and dough strength traits on selected hard spring wheat samples. Research will be conducted on the rapid characterization of protein extracts from wheat and flour by testing two different protein extraction methods. Variation in free asparagine content in wheat lines will be measured. The efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat, specifically protein composition and amino acid profiles, will be determined as a function of dough strength characteristics. To develop optimized methodologies for the extraction of beta-glucan from oats, samples of oats will be obtained from replicated plots grown in diverse environments over several years. The newly developed methods for the analysis of soluble fiber quality will be used to determine how environment might affect the quality and concentration of beta-glucan, especially in newly developed high beta-glucan cultivars. Beta-glucan effects on milling quality of oats will also be tested. Because beta- glucan is in the cell walls of oats, it is thought to provide a strengthening effect, which reduces groat breakage during dehulling. However, high oil in oats also seems to prevent groat breakage, and currently breeders have been unable to separate the traits of high oil and high beta-glucan in oats. A collaborating genetics program has developed populations of recombinant inbred lines from oat crosses designed to be segregating for high oil and high beta-glucan. We will evaluate the phenotypes of these lines and test their behavior during dehulling to determine the relative roles of oil and beta-glucan in preventing groat breakage during oat milling. The Hard Red Spring Wheat Quality Laboratory (WQL) analyzed physical and biochemical quality traits of approximately 3000 samples of hard spring and durum wheat. Samples were submitted by private and public wheat breeders involved in wheat germplasm improvement and by scientists involved in gene mapping to identify quantitative trait loci associated with end-use quality traits. The WQL provided over 40 different tests related to wheat kernel characteristics, milling performance, and flour, semolina, dough, baking, and spaghetti processing quality. In cooperation with the Wheat Quality Council (WQC), eleven experimental lines of hard spring wheat that were grown at up to 5 locations were evaluated along with the check cultivars, Glenn and Elgin-ND. The WQL tested each line for kernel, milling, flour, dough, and bread-baking quality traits, coordinated the baking quality test results from 13 independent public and private testing laboratories, and analyzed the data. Results were published, presented, and discussed at the annual meeting of the WQC. Interactions with the WQC serve as a means to obtain industry feedback on the milling and baking quality traits of advanced experimental lines of wheat that are considered for release into commercial production. We continued research on the effects of protein molecular weight distribution (MWD) on bread-making characteristics of hard spring wheat. Approximately, 300 samples harvested from Minnesota in 2013 were analyzed by size-exclusion high performance liquid chromatography (SE-HPLC). The sodium dodecyl sulfate (SDS) buffer soluble and insoluble protein fractions that were extracted from those samples were also evaluated by UV spectroscopy. The data obtained from this research will be evaluated to develop prediction models of quality traits. Waxy wheat is unique since its starch is primarily composed of amylopectin. One hundred ninety eight lines of hard winter waxy wheat were analyzed by SE-HPLC. The data will be analyzed to investigate the association between protein MWD and breadmaking quality in waxy wheat. Wheat kernel vitreousness is an important grading characteristic in the US. Research was performed to determine the possible association between kernel vitreousness and protein MWD, and breadmaking quality. Data analysis indicated that high molecular weight polymeric proteins could have a significant association with kernel vitreousness. The changes in protein MWD were analyzed during pasta cooking using SE-HPLC. The data will be analyzed to determine the relationships with pasta cooking characteristics. Solvent retention capacity (SRC) is a test to evaluate flour baking quality. SRC has not been tested for evaluation of breadmaking quality of hard spring wheat. We analyzed SRC and protein MWD of 10 hard spring wheat genotypes grown at 10 locations in North Dakota. Data analysis is ongoing to determine the potential association between SRC parameters and protein MWD and quality traits. The segregation of low asparagine wheat genotypes is important to reduce acrylamide content in baking products. Free asparagine concentration is being analyzed for 132 hard spring wheat samples grown in Minnesota and the data will be analyzed to investigate the variation in free asparagine for spring wheat lines grown in Minnesota. Accomplishments 01 Development of improved wheat germplasm. Wheat producers, milling and baking industries, and overseas customers require high standards in the quality of wheat to meet their evolving needs. The Hard Red Spring and Durum Wheat Quality Laboratory contributed wheat end-use quality data that helped lead to the development of improved wheat germplasm and subsequent release of new cultivars of spring and durum wheat bred for commercial production. Specifically, this work contributed to the release of two hard spring wheat cultivars, �Rollag� by University of Minnesota and 'Advance' by South Dakota State University, in 2014. Such new releases provide value to both farmers and industry through improved productivity or end-use quality. 02 Effect of protein on flour dough thermal properties. Flour bread making quality is very important for wheat marketing in domestic and international trade, and thermal tolerance of dough to heating has been shown to influence flour bread making quality. ARS and NDSU researchers in Fargo, ND studied the influence of flour proteins on properties of hard red spring wheat dough during heating and bread making, and found that less soluble polymeric proteins had a significant and positive effect on dough thermal tolerance and strength, and consequently, bread loaf volume. This work expands our knowledge of the function of proteins in bread making and will be useful for the evaluation of wheat and flour quality of hard spring wheat. 03 Effect of wheat kernel size on protein, milling yield, and baking quality. Optimization of flour yield and quality is important in the wheat milling industry, but flour yield from roller mills can be reduced by having a mixture of wheat seeds with different sizes. ARS and NDSU researchers in Fargo, ND determined how seed size and experimental mill type is associated with milling and bread making quality. When compared to large seeds, small seeds have a greater level of high molecular weight polymeric proteins that can contribute to the enhancement of flour bread making quality, while still having poor milling quality. This information will be helpful for segregation and improvement of hard spring wheat for milling and bread making quality. 04 Occurrence of mycotoxins in hard red spring wheat affected by Fusarium head blight. Deoxynivalenol (DON) is a mycotoxin found in wheat with a disease known as Fusarium head blight. Deoxynivalenol-3-glucoside (D3G) is a type of "masked mycotoxin." ARS and NDSU researchers in Fargo, ND evaluated the DON and D3G content of hard spring wheat samples collected from the 2011 and 2012 crop surveys of the USA. Wheat samples showed a significant difference in DON and D3G content according to growing region. This work provides more accurate data regarding total mycotoxin levels in hard spring wheat. The data should be useful for the segregation of hard spring wheat for total mycotoxin level in the milling and baking industry. 05 Mycotoxin in hard red spring wheat affected by Fusarium head blight in Minnesota. The "masked" mycotoxin deoxinyvalenol-3-glucoside (D3G) is important from a food safety point of view, since it can harm human and animal health. Unfortunately, no data regarding D3G has been available for hard spring wheat genotypes. ARS and NDSU researchers in Fargo, ND and researchers in University of Minnesota determined the level of mycotoxins, including D3G, in hard spring wheat varieties grown in Minnesota. This work provides more precise information on total mycotoxin content of hard spring wheat varieties grown at diverse locations in Minnesota. This information will help identify hard spring wheat varieties with stably low mycotoxin content across growing locations and will thus facilitate further development of wheat with low mycotoxin levels. 06 Addition of glucose oxidase for the improvement of refrigerated dough quality. Refrigerated dough encompasses a wide range of products and is a very popular choice for consumers. Two of the greatest problems that occur during refrigerated dough storage are water weeping from dough (dough syruping) and decreased dough baking quality. ARS and NDSU researchers in Fargo, ND examined the effect of an enzyme called glucose oxidase on refrigerated dough quality. The optimum level of glucose oxidase was 10 ppm, at which level dough syruping was reduced, while dough bread-making quality was maintained during refrigerated storage. This information will help the promotion of hard spring wheat flour for the production of refrigerated dough with improved quality.

Impacts
(N/A)

Publications

• Simsek, S., Ohm, J.-B., Lu, H., Rugg, M., Berzonsky, W., Alamri, M.S., Mergoum, M. 2014. Effect of pre-harvest sprouting on physicochemical properties of starch in wheat. Foods. 3:194-207.
• Ovando-Martinez, M., Ozsisli, B., Anderson, J., Whitney, K., Ohm, J.-B., Simsek, S. 2013. Analysis of deoxynivalenol and deoxynivalenol-3-glucoside in hard red spring wheat inoculated with Fusarium graminearum. Toxins. 5:2522-2532.
• Simsek, S., Ovando-Martinez, M., Ozsisli, B., Whitney, K., Ohm, J. 2013. Occurrence of deoxynivalenol and deoxynivalenol-3-glucoside in hard red spring wheat grown in the USA. Toxins. 5:2656-2670.
• Simsek, H., Kasi, M., Ohm, J.-B., Blonigen, M., Khan, E. 2013. Bioavailable and biodegradable dissolved organic nitrogen in activated sludge and trickling filter wastewater treatment plants. Water Research. 47:3201-3210.
• Simsek, S., Ohm, J., Lu, H., Rugg, M., Berzonsky, W., Alamri, M., Mergoum, M. 2013. Effect of pre-harvest sprouting on physicochemical changes of proteins in wheat. Journal of the Science of Food and Agriculture. 94:205- 212.
• Tsilo, T.J., Hareland, G.A., Ohm, J., Anderson, J.A. 2013. Polymeric proteins and their association with grain yield in hard red spring wheat lines. Euphytica. 194:187-196.
• Cremer, J.E., Liu, L., Bean, S., Ohm, J., Tilley, M., Wilson, J.D., Kaufman, R.C., Vu, T.T., Gilding, E.K., Godwin, I., Wang, D. 2014. Impacts of kafirin allelic diversity, starch content, and protein digestibility on ethanol conversion efficiency in grain sorghum. Cereal Chemistry. 91(3) :218-227.
• Sivapragasam, N., Thavarajah, P., Ohm, J.-B., Khaitsa, M., Thavarajah, D. 2014. Novel starch based nano scale enteric coatings from soybean meal for colon-specific delivery. Carbohydrate Polymers. 111:273-279.
• Sivapragasam, N., Thavarajah, P., Ohm, J.-B., Thavarajah, D. 2014. Enzyme resistant carbohydrate based micro-scale materials from sugar beet (Beta vulgaris L.) pulp for food and pharmaceutical applications. Bioactive Carbohydrates and Dietary Fibre. 3:115-121.

Progress 10/01/12 to 09/30/13

Outputs
Progress Report Objectives (from AD-416): The ultimate goal is to enhance the quality of U.S. wheat for domestic and overseas markets. Objectives are to: identify milling, baking, and pasta processing traits of experimental and advanced lines of hard spring and durum wheat that impact end-use quality and the overall development and release of new commercial cultivars of wheat; and identify specific biochemical components in wheat that are associated with end-use baking quality traits. More specifically, the latter objective includes: to develop rapid methods to characterize proteins, which are initially important for predicting end-use quality; to determine the variation in free asparagine content in wheat, which is an important precursor for the formation of carcinogenic acrylamide during bread baking and toasting; and to identify the efficacy and impact of sulfur fertilization on end- use baking quality of hard spring wheat. Approach (from AD-416): Approximately 4000 hard spring and 1000 durum wheat experimental lines will be submitted by Federal, State, and private breeders, and special interest groups on an annual basis and evaluated for end-use quality. Wheat will be obtained from different generations of development in specific breeding programs, which will include early-generation, preliminary, and advanced lines and include commercial cultivars as controls. Measureable end-use quality traits include test weight, size, seed weight, sprout damage, physical evidence of disease, and protein and ash content; milling performance as a function of flour and semolina yield; flour and semolina color, protein, and ash content; dough strength properties; bread baking quality; and pasta processing quality. The Fargo Genotyping Laboratory will identify quantitative trait loci regions associated with end-use quality, primarily kernel and dough strength traits on selected hard spring wheat samples. Research will be conducted on the rapid characterization of protein extracts from wheat and flour by testing two different protein extraction methods. Variation in free asparagine content in wheat lines will be measured. The efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat, specifically protein composition and amino acid profiles, will be determined as a function of dough strength characteristics. The Hard Red Spring Wheat Quality Laboratory (WQL) analyzed physical and biochemical quality traits of approximately 3000 samples of hard spring and durum wheat. Samples were submitted by private and public wheat breeders involved in wheat germplasm improvement and by scientists involved in gene mapping to identify quantitative trait loci associated with end-use quality traits. The WQL provided over 40 different tests related to wheat kernel characteristics, milling performance, and flour, semolina, dough, baking, and spaghetti processing quality. A test to measure bread crumb firmness that is expected to enhance the objective evaluation of bread quality was added to the existing quality tests. In cooperation with the Wheat Quality Council (WQC), fourteen experimental lines of hard spring wheat that were grown at up to 5 locations were evaluated along with the check cultivar, Glenn. The WQL tested each line for kernel, milling, flour, dough, and bread-baking quality traits, coordinated the baking quality test results from 13 independent public and private testing laboratories, and analyzed the data. Results were published, presented, and discussed at the annual meeting of the WQC. Interactions with the WQC serve as a means to obtain industry feedback on the milling and baking quality traits of advanced experimental lines of wheat that are considered for release into commercial production. We found that polymeric proteins in SDS buffer soluble and insoluble fractions significantly affected the bread-making characteristics of hard spring wheat. We analyzed polymeric proteins from durum wheat samples and are currently investigating their relationship with durum quality traits. Also, we are investigating the effect of genetic background and growing environment on variation of polymeric proteins and quality characteristics in hard spring and durum wheat. UV spectroscopy has been continuously tested for the rapid characterization of proteins extracted from hard spring and durum wheat samples. SDS buffer soluble and insoluble protein fractions have been extracted from hard spring and durum wheat samples and scanned at wavelengths between 190 and 350 nm using a UV spectrophotometer. We are currently examining possible correlations between UV absorbance data and durum quality traits. The utility of near infrared spectroscopy for determination of flour polymeric protein content in hard spring and durum wheat also will be assessed. The segregation of low asparagine wheat genotypes is important to reduce acrylamide content in baking products. Free asparagine concentration was analyzed for 75 hard spring wheat lines grown at 3 locations and the data is currently being analyzed to investigate the effects of genotype and location. Accomplishments 01 Development of improved wheat germplasm. Wheat producers, milling and baking industries, and overseas customers require high standards in the quality of wheat to meet their evolving needs. ARS researchers in Fargo, ND contributed wheat end-use quality data that helped lead to the development of improved wheat germplasm and subsequent release of new cultivars of spring and durum wheat bred for commercial production. Such new releases provide value to both farmers and industry through improved productivity or end-use quality. 02 Polymeric proteins and their association with grain yield in hard red spring wheat. One of the problems faced by wheat breeding programs is that varieties with high grain yield tend to have low protein content and poor end-use quality. Large protein aggregates known as high molecular weight polymeric proteins contribute to enhancing bread- making quality. In cooperation with wheat breeders from the University of Minnesota, ARS researchers in Fargo, ND found that an increase of polymeric proteins could improve bread-making quality without decreasing grain yield in hard spring wheat. This information provides breeders with knowledge needed to improve selection of wheat genotypes for improved grain yield without loss in bread-making quality. 03 Genetic mapping analysis of bread-making quality traits in hard spring wheat. Bread-making quality is an important characteristic considered before release of new spring wheat varieties, but measuring bread- making quality is both time and labor intensive. Screening using genetic markers has the potential to aid in this process. A mapping population derived from the wheat lines BR34 and Grandin by ARS researchers in Fargo, ND was used to identify genetic markers for bread- making quality traits in hard spring wheat. A total of 31 quantitative trait loci (QTL) were identified for quality traits. These QTL will be useful in segregating wheat lines for breadmaking quality during early generations of breeding. 04 Variability in arabinoxylan, xylanase activity, and xylanase inhibitor levels in hard spring wheat. Arabinoxylans, xylanase, and xylanase inhibitors have an important role in many cereal food processing applications. Xylanase is an enzyme needed to break arabinoxylans into smaller molecules, and optimum xylanase activity in wheat is beneficial to breadmaking. ARS and NDSU researchers in Fargo, ND examined the effects of genotype, growing location, and their interaction on arabinoxylans, apparent xylanase activity, and apparent xylanase inhibition activity of Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibiting protein for six hard red and six hard white spring wheats. TAXI was identified as a reliable parameter for segregating wheat genotypes with varying xylanase activity. This information will help segregation of wheat lines that have different levels of xylanase activity in wheat breeding. 05 Effect of a high molecular weight glutenin subunit gene on the quality of durum wheat. High molecular weight glutenin subunits Glu-D1d are proteins associated with good bread-making quality that exist in bread wheat, but not in durum wheat. ARS researchers in Fargo, ND transferred the gene encoding Glu-D1d to durum and found that this glutenin gene enhanced the bread-making quality characteristics of the durum lines. Most durum lines with this glutenin gene also had a lower free asparagine content (free asparagine is a precursor needed to form carcinogenic acrylamide during baking) than those without it. This information may be useful to breeders interested in development of a dual-use durum wheat for pasta- and bread-making that could contribute to diversifying the utilization of durum wheat.

Impacts
(N/A)

Publications

• Simsek, S., Whitney, K., Ohm, J. 2013. Analysis of Cereal Starches by High Performance Size Exclusion Chromatography. Journal of Food Analytical Methods. 6(1): 181-190.
• Mendis, M., Ohm, J., Delcour, J., Gebruers, K., Meinhardt, S., Simsek, S. 2013. Variability in Arabinoxylan, Xylanase activity and Xylanase inhibitor levels in hard spring wheat. Cereal Chemistry. 90:240�248.

Progress 10/01/11 to 09/30/12

Outputs
Progress Report Objectives (from AD-416): The ultimate goal is to enhance the quality of U.S. wheat for domestic and overseas markets. Objectives are to: identify milling, baking, and pasta processing traits of experimental and advanced lines of hard spring and durum wheat that impact end-use quality and the overall development and release of new commercial cultivars of wheat; and identify specific biochemical components in wheat that are associated with end-use baking quality traits. More specifically, the latter objective includes: to develop rapid methods to characterize proteins, which are initially important for predicting end-use quality; to determine the variation in free asparagine content in wheat, which is an important precursor for the formation of carcinogenic acrylamide during bread baking and toasting; and to identify the efficacy and impact of sulfur fertilization on end- use baking quality of hard spring wheat. Approach (from AD-416): Approximately 4000 hard spring and 1000 durum wheat experimental lines will be submitted by Federal, State, and private breeders, and special interest groups on an annual basis and evaluated for end-use quality. Wheat will be obtained from different generations of development in specific breeding programs, which will include early-generation, preliminary, and advanced lines and include commercial cultivars as controls. Measureable end-use quality traits include test weight, size, seed weight, sprout damage, physical evidence of disease, and protein and ash content; milling performance as a function of flour and semolina yield; flour and semolina color, protein, and ash content; dough strength properties; bread baking quality; and pasta processing quality. The Fargo Genotyping Laboratory will identify quantitative trait loci regions associated with end-use quality, primarily kernel and dough strength traits on selected hard spring wheat samples. Research will be conducted on the rapid characterization of protein extracts from wheat and flour by testing two different protein extraction methods. Variation in free asparagine content in wheat lines will be measured. The efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat, specifically protein composition and amino acid profiles, will be determined as a function of dough strength characteristics. The Hard Red Spring and Durum Wheat Quality Laboratory (WQL) analyzed the physical and biochemical quality traits on approximately 3000 samples of hard spring and durum wheat and provided over 40 different quality tests. In cooperation with the Wheat Quality Council (WQC), ARS scientists evaluated fourteen experimental lines of hard spring wheat that were grown at up to 4 locations along with the check cultivar, Glenn. The WQL tested each line for kernel, milling, flour, dough, and bread-baking quality traits; coordinated the baking quality test results from 12 independent public and private testing laboratories; and analyzed the data and published the results that were presented and discussed at the annual meeting of the WQC in 2012. Polymeric proteins in SDS buffer soluble and insoluble proteins were found to have contrasting effects (negative effect for soluble proteins and positive effect for insoluble proteins) on bread-making traits. The SE-HPLC with a micro bore column previously appeared to enhance the speed of analysis of polymeric proteins in wheat and therefore this line of investigation was continued to confirm whether the micro bore column is useful for quality evaluation. Specifically, UV absorbance data of proteins in wheat was collected at 195, 214, 275, and 280 nm using SE- HPLC; proteins were observed to show different responses at those wavelengths. The UV absorbance data will be investigated to determine if it�s useful for quality evaluation. We also tested UV spectroscopy for rapid characterization of protein extracts. The SDS buffer soluble and insoluble protein fractions were extracted from hard spring wheat flour samples and were scanned at wavelengths between 190 and 350 nm. Absorbance at specific wavelengths was identified to have significant correlations with flour bread making properties. Research will continue to determine if the absorbance data are useful for quality evaluation. Near- and mid- infrared spectroscopy will also be tested for suitability for characterization of protein extracts. Wheat asparagine concentration is known to influence carcinogenic acrylamide formation during bread baking and toasting, and segregation of low asparagine wheat genotypes is important to reduce acrylamide content in baking products. Pre-harvest sprouting increases free asparagine concentration in wheat. Free asparagine in sprouted wheat samples of 12 hard red and 12 white spring wheat genotypes grown at 3 locations was analyzed by GC-MS. Hard spring wheat genotypes with tolerance to pre- harvest sprouting and low asparagine concentration can be segregated. Asparagine content was analyzed for durum translocation lines that carry chromosome segments for high-molecular-weight glutenin subunits from the D genome of common wheat. These data are currently being analyzed to investigate any genetic association of high-molecular-weight glutenin subunits and asparagine concentration. Asparagine concentration is also currently being analyzed for 75 hard spring wheat lines grown at 3 locations to investigate the effect of genotype and growing location. Accomplishments 01 Development of improved wheat germplasm. Wheat producers, milling and baking industries, and overseas customers require high standards in the quality of wheat to meet their evolving needs. The ARS Hard Red Spring and Durum Wheat Quality Laboratory (WQL) in Fargo, ND contributed wheat end-use quality data that helped lead to the development of improved whe germplasm and subsequent release of new cultivars of spring and durum wheat bred for commercial production. Four hard spring wheat cultivars, for which quality characteristics were evaluated in the WQL, were releas in 2011-12. Two cultivars, Forefront and Advance, were released by the wheat breeding program at South Dakota State University and two cultivar Tom and Sabin, by the wheat breeding program at the University of Minnesota. 02 Effect of pre-harvest sprouting on wheat proteins and free asparagine concentration. Rainfall before wheat harvest can cause pre-harvest sprouting (PHS) of the wheat kernel, which results in degradation of whe quality. Work conducted at the ARS Wheat Quality Laboratory in Fargo, ND characterized specific biochemical changes that occur with PHS damage an determined that genetic selection may be useful for mitigating damage fr PHS. This information provides breeders and millers with the knowledge needed to improve selection of wheat genotypes for improved PHS toleranc and bread-making quality. 03 Genetic markers associated with endosperm proteins and end-use quality traits of hard red spring wheat breeding lines. This research was performed in cooperation with wheat breeders from the University of Minnesota to identify genetic markers of endosperm polymeric proteins th are known to greatly affect flour gluten characteristics and thus breadmaking quality. A population of 139 recombinant inbred lines was evaluated in three environments. Eleven genetic markers linked to endosperm polymeric proteins were identified. The DNA markers linked to these polymeric proteins will be useful for breeders in selecting wheat lines that have strong gluten strength characteristics during early generations of breeding. 04 Prediction of flour breadmaking characteristics. The Mixolab is an instrument that provides data on flour dough thermal characteristics and mixing properties. Use of data from this instrument can be used in mathematical models to predict the quality of wheat varieties under development for commercial purposes. In cooperation with North Dakota State University, ARS scientists in Fargo, ND used Mixolab data to devel highly predictive calibration models for breadmaking parameters in 30 ha red spring wheat flour samples. The development of robust prediction models will facilitate the objective evaluation of wheat flour for breadmaking quality.

Impacts
(N/A)

Publications

• Tsilo, T.J., Ohm, J., Hareland, G.A., Chao, S., Anderson, J.A. 2011. Quantitative Trait Loci influencing endosperm proteins and end-use quality traits of Hard Red Spring Wheat breeding lines. Czech Journal of Genetics and Plant Breeding. 47:S190-S195.
• Simsek, S., Whitney, K., Ohm, J., Mergoum, M. 2011. Refrigerated dough quality of hard red spring wheat: Effect of genotype and environment on dough syruping and arabinoxylan production. Cereal Chemistry. 88(5):445- 450.

Progress 10/01/10 to 09/30/11

Outputs
Progress Report Objectives (from AD-416) The ultimate goal is to enhance the quality of U.S. wheat for domestic and overseas markets. Objectives are to: identify milling, baking, and pasta processing traits of experimental and advanced lines of hard spring and durum wheat that impact end-use quality and the overall development and release of new commercial cultivars of wheat; and identify specific biochemical components in wheat that are associated with end-use baking quality traits. More specifically, the latter objective includes: to develop rapid methods to characterize proteins, which are initially important for predicting end-use quality; to determine the variation in free asparagine content in wheat, which is an important precursor for the formation of carcinogenic acrylamide during bread baking and toasting; and to identify the efficacy and impact of sulfur fertilization on end- use baking quality of hard spring wheat. Approach (from AD-416) Approximately 4000 hard spring and 1000 durum wheat experimental lines will be submitted by Federal, State, and private breeders, and special interest groups on an annual basis and evaluated for end-use quality. Wheat will be obtained from different generations of development in specific breeding programs, which will include early-generation, preliminary, and advanced lines and include commercial cultivars as controls. Measureable end-use quality traits include test weight, size, seed weight, sprout damage, physical evidence of disease, and protein and ash content; milling performance as a function of flour and semolina yield; flour and semolina color, protein, and ash content; dough strength properties; bread baking quality; and pasta processing quality. The Fargo Genotyping Laboratory will identify quantitative trait loci regions associated with end-use quality, primarily kernel and dough strength traits on selected hard spring wheat samples. Research will be conducted on the rapid characterization of protein extracts from wheat and flour by testing two different protein extraction methods. Variation in free asparagine content in wheat lines will be measured. The efficacy and impact of sulfur fertilization on end-use baking quality of hard spring wheat, specifically protein composition and amino acid profiles, will be determined as a function of dough strength characteristics. This project continues research from Project 5442-43440-008-00D Enhance Hard Spring and Durum Wheat Quality and Utilization. The Hard Red Spring Wheat Quality Laboratory (WQL) analyzed the physical and biochemical quality traits on over 4000 samples of hard spring and durum wheat that were submitted by private and public wheat breeders involved in improving wheat germplasm and by scientists involved in gene mapping to identify chromosome regions harboring genes that influence end-quality traits. The WQL, in cooperation with the Wheat Quality Council (WQC), evaluated twelve experimental lines of hard spring wheat that were grown at up to 5 locations. The WQL tested each line for kernel, milling, flour, dough, and baking quality traits; coordinated the baking quality test results from 9 independent public and private testing laboratories; and analyzed the data and published the results that were presented and discussed at the annual meeting of the WQC in 2010. WQL interaction with the WQC serves to provide industry feedback on milling and baking quality traits of advanced experimental lines of wheat that are considered for release into commercial production. Rapid characterization of polymeric proteins in flour is important for wheat breadmaking quality evaluation. We tested a micro bore column for analysis of polymeric proteins that were extracted from hard spring wheat flour. The procedure using a microbore column appears to enhance the speed of analysis of polymeric protein for wheat quality evaluation with decreased consumption of hazardous organic solvents. We also tested UV spectroscopy for rapid characterization of protein extracts and identified wavelengths of which absorbance data might be important for quality evaluation based on their correlations with flour breadmaking properties. Since free asparagine is a precursor for the formation of carcinogenic acrylamide during bread baking and toasting, segregation of wheat lines that have low asparagine concentration is important. We tested two solutions to extract free amino acids in wheat for analysis of asparagine using a gas chromatography system and selected a hot ethanol solution due to the higher stability in amino acid concentration during extraction. We analyzed 48 wheat lines grown in two locations in ND for free amino acids. The mixolab is a new empirical instrument that measures dough rheology characteristics under thermal constraints as well as mixing properties. The mixolab profile was divided into six different stages, and torque measurements of individual stages were modeled by nonlinear curve fitting using a combination of genetic algorithms and multidimensional unconstrained nonlinear minimization. Novel parameters were extracted from the fitted equations. The mixolab parameters are being further tested to determine if they can be employed to develop prediction models of breadmaking characteristics in cooperation with the wheat breeding program at South Dakota State University. The new mixolab parameters will result in improvement of objective evaluations of flour dough thermal characteristics that also will be helpful for evaluation of flour bread making quality. Accomplishments 01 Development of improved wheat germplasm. Wheat producers, milling and baking industries, and overseas customers require high standards in the quality of wheat to meet their evolving needs. ARS researchers in Fargo ND contributed wheat end-use quality data that helped lead to the development of improved wheat germplasm and subsequent release of new cultivars of spring and durum wheat bred for commercial production. The provided over 40 different tests related to the physical and biochemical quality traits of the wheat kernel and related milling performance, flou semolina, dough, baking, and spaghetti processing on over 4000 samples o hard spring, and durum wheat lines that were submitted by public and private wheat breeders and cooperating scientists. As a result, new spring wheat cultivars were released in 2009/10 that provide farmers, industry, and consumers with high quality wheat. 02 Quality of refrigerated dough made from hard red spring wheat. Products obtained from refrigerated doughs account for greater than $1.7 billion per year in sales in the US, and they are one of the fastest growing segments of the ready-to-use, grain-based food industry. Refrigerated storage of bread dough products can result in undesirable dough syruping (water separation). ARS researchers in Fargo, ND, in collaboration with North Dakota State University, investigated the effects of wheat cultiva and growing environment on dough syruping during refrigerated storage in relation to xylanase activity (a plant cell wall or sugar degrading enzy activity) in hard red spring wheat. They discovered that growing environment had a greater impact on enzyme activity and dough syruping than cultivar of wheat. Some cultivars were consistently lower in appare enzyme activity and dough syruping across all growing environments. The results indicate that certain cultivars grown in relatively dry environments across the prime hard red spring wheat growing region can b used in refrigerated dough formulations. This information will help millers and wheat breeders segregate cultivars for refrigerated dough production.

Impacts
(N/A)

Publications

• Sandhu, H., Manthey, F.A., Simsek, S., Ohm, J. 2011. Comparison Between Potassium Bromate and Ozone as Flour Oxidants in Breadmaking. Cereal Chemistry. 88(1):103�108.
• Simsek, S., Whitney, K.L., Ohm, J., Anderson, J., Mergoum, M. 2011. Refrigerated Dough Quality: Effect of Environment and Genotypes of Hard Red Spring Wheat. Journal of Food Science. 76:S101-S107.
• Caffee-Treml, M., Glover, K.D., Krishnan, P., Hareland, G.A. 2010. Variability and Relationships Among Mixolab, Mixograph, and Baking Parameters Based on Multi-Environment Spring Wheat Trials. Cereal Chemistry. 87:574-580.
• Ohm, J., Klindworth, D.L., Hareland, G.A., Faris, J.D., Elias, M., Xu, S.S. 2010. Variation in Kernel Characteristics and Protein Molecular Weight Distribution of Langdon Durum�Wild Emmer Wheat Chromosome Substitution Lines. Journal of Cereal Science. 52:207-214.
• Ohm, J., Hareland, G.A., Simsek, S., Seabourn, B.W., Maghirang, E.B., Dowell, F.E. 2010. Molecular weight distribution of proteins in hard red spring wheat: Relationship to quality parameters and intra-sample uniformity. Cereal Chemistry. 87(6):553�560.
• Liu, Y., Ohm, J., Hareland, G.A., Wiersma, J., Kaiser, D. 2011. Sulfur, Protein Size Distribution, and Free Amino Acids in Flour Mill Streams and Their Relationship to Dough Rheology and Breadmaking Traits. Cereal Chemistry. 88(2):109-116.
• Glover, K.D., Rudd, J.C., Devkota, R.N., Hall, R.G., Jin, Y., Osborne, L.E. , Ingemansen, J.A., Richertsen, J.R., Baltensperger, D.D., Hareland, G.A. 2010. Registration of "Brick" Wheat. Journal of Plant Registrations. 4:22- 27.
• Garvin, D.F., Hareland, G.A., Gregoire, B.R., Finley, J.W. 2011. Impact of wheat grain selenium content variation on milling and bread baking. Cereal Chemistry. 88(2):195-200.
• Hall, M.D., Rohrer-Perkins, W., Griffey, C.A., Liu, S.Y., Thomason, W.E., Abaye, A.O., Bullard-Schilling, P.G., Gundrum, P.G., Fanelli, J.K., Chen, J., Brooks, W.S., Seago, J.E., Will, B.C., Hokanson, E.G., Behl, H.D., Pitman, R.M., Kenner, J.C., Vaughn, M.E., Corbin, R.A., Dunaway, D.W., Lewis, T.R., Starner, D.E., Gulick, S.A., Beahm, B.R., Whitt, D.L., Lafferty, J.B., Hareland, G.A. 2011. Registration of �Snowglenn� Winter Durum Wheat. Journal of Plant Registrations. 5(1):1-6.
• Tsilo, T.J., Hareland, G.A., Chao, S., Anderson, J.A. 2011. Genetic Mapping and QTL Analysis of Flour Color and Milling Yield Related Traits Using Recombinant Inbred Lines in Hard Red Spring Wheat. Crop Science. 51:237-246.
• Tsilo, T.J., Linkert, G.L., Hareland, G.A., Anderson, J.A. 2011. Registration of the MN98550/MN99394 Wheat Recombinant Inbred Mapping Population. Journal of Plant Registrations. 5(2):257-260.