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Amino acid analysis of GEM germplasm M. Paul Scott USDA-ARS Ames, Iowa 50011 [email protected] U.S. Corn Uses National Corn Growers Association, 2004 Annual value: $25 Billion Feed/residual Export Ethanol HFCS Starch Sweeteners Cereal Alcohol Seed Impact of increased ethanol production Corn Prices Corn Acres Soybean price Gov’t Payments Livestock income Overall Farm income N, P Lost to water Soil Erosion Up Up Down Down Slight decrease Up Up Up Source: WRI policy note,2006 Marshall & Greenhalgh Annual meat consumption, per capita (Source: FAO) 140 120 80 United States 60 World average 40 20 Year 1997 1993 1989 1985 1981 1977 1973 1969 1965 0 1961 kg 100 Food and Fuel Petroleum Grain hydrocarbons Starch Protein Oil Transportation Feed Oilseeds Protein Oil Food Food and Fuel Grain Starch Protein Oil Petroleum Oilseeds Protein Oil 3.9 Billion Gallons EtOH in 2005 13% of Corn crop Transportation 146 Billion Gallons Gasoline used/year in US Feed Food A solution Petroleum Transportation Lignocellulosic Biomass Grain Starch Protein Oil Feed Grain + Ethanol Co-products: DDGs Oilseeds Protein Oil Food What corn traits are important? • Yield (insect, disease, drought …) • N use efficiency • What about quality traits? Grain Ethanol Production Protein Fiber Fat Starch 10% 3% 4% 75% Corn Protein is concentrated by ethanol production! Problems are amplified! Starch 2.7 gal/bu $4.32/bu Protein, Fiber, Fat EtOH Co-products: DDGS, Solubles 18 lb/bu $0.68/bu Protein Fiber Fat Starch 30% 10-50% 10-20% 5% Seed Fractionation Osborn, 1924 Water Albumins Salt Globulins Alcohol Prolamins “Zeins” Base Glutelins Seed Proteins Embryo-vegetative Endosperm-storage Zeins SDS-PAGE Coomassie Blue Stained Zeins • Zeins are so abundant that they have a large impact on protein quality of grain • Zeins are concentrated in ethanol coproducts so their impact is even greater • Zeins are nutritionally poor for nonruminants – They are poorly digested – They are deficient in certain essential amino acids Amino acid levels Corn vs. Egg 2.5 2 Maize Soy 1.5 1 0.5 0 Lys His Arg Asx Thr Ser Glx Pro Gly Ala Cys Val Met Ile Leu Tyr Phe Trp Nutritionally limiting amino acids Methionine is the limiting AA in Poultry diets Lysine is limiting in poultry and swine diets, but can be supplemented inexpensively Tryptophan is limiting in swine diets What corn traits are important? • Yield (insect, disease, drought …) • N use efficiency For Ethanol: •Fermentable starch •Protein quality •Available P Amino acid/ protein For non-ruminant feed: •Protein content •Protein quality •Available P Absolute amino acid level Questions about biofuel coproducts • How are differences in grain composition reflected in co-product composition? • How will co-products be improved by processing? • Can yeast components be used to complement grain components in coproducts? Evaluation of amino acid content of GEM germplasm • About 80 GEM accessions / year selected for agronomics and past AA performance. • Good lines are re-tested in subsequent years • Lys, Met, Trp, Index (Lys,Met,Trp) • Checks: – B101 (high Met) – B45o2 (high Lys, Trp) – B73xMo17, B73, Mo17, B45 Microbial assay for Met Low High Met Concentration Evaluation of amino acid composition of GEM germplasm M 0.20 0.15 0.10 M/protein 0.020 0.015 0.010 Two year Evaluation Evaluation of amino acid composition of GEM germplasm 0.016 0.20 M 0.16 0.14 0.12 0.10 0.08 0.014 M/protein 0.18 0.012 0.01 0.008 0.006 Four year evaluation Highlights of the GEM AA program • DKXL212:N11a-139 Index was significantly higher than B73xMo17 in 4 yr tests. • CHIS740:S1411a-783-2 Met/protein was not significantly different that hi met check and significantly higher than other checks in 4 yr tests • Seven XL370 and XL380 derived lines were not different than high met check in 2 yr evaluations. Recommendations • Evaluation of hybrids • Selection within GEM “lines” • Development of synthetics for recurrent selection Divergent selection for amino acid content Long-term Selection for protein content Advantages: Differences are magnified by selection in both directions Generate unique material to answer biological questions Source: Dudley and Lambert (2004), Plant Breeding Reviews 24, 7979-110 Recurrent Selection Intermating of Selections BS31HM C1 Selections (5 ears) BS31HM Selections (5 ears) BS31 Evaluation of 50 ears BS31HM C1 BS31HM C2 Divergent selection for AA Content BS31 0.13 0.15 0.12 0.14 0.11 0.13 M (g/100g) M (g/100g) BS11 0.1 0.09 0.08 0.07 0.11 0.1 0.09 0.06 0.08 -4 -3 -2 -1 0 1 Cycle 2 3 4 -4 0.013 0.013 0.012 0.012 0.011 0.011 W (g/100g) W (g/100g) 0.12 0.01 0.009 0.008 0.007 -3 -2 -1 0 1 Cycle -3 -2 -1 2 3 4 2 3 4 0.01 0.009 0.008 0.007 0.006 0.006 -4 -3 -2 -1 0 1 Cycle 2 3 4 -4 0 1 Cycle Acknowledgements • Mike Blanco and the GEM team • Merinda Struthers • Funding from USDA-ARS project funds Thanks for your attention!