Annual Progress Report on Malting Barley Research March, 2007

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A-Chain A-Chain A-Chain B-Chain C-Chain Figure 4. Portion of an idealized starch (amylopectin) molecule. Gray circles show glucose residues that would be removed by beta-amylase in the preparation of betalimit dextrin substrate. Striped circles show residues within the beta-limit dextrin that would be susceptible to beta-amylase only after initial treatment with alpha-amylase. Methods and Results. In the first portion of this study high performance size exclusion chromatography (HPSEC) was used to compare the change in the molecular weight of the beta-limit dextrin (HPSEC) substrate to the loss in iodine binding when incubated with alpha-amylase. As shown in figure 5, there was more than a 7-fold reduction in molecular weight within the first min of reaction, while iodine binding was only lost slowly. The absorbance at 12 min corresponds approximately to the end-point of the manual method. These results suggest that fragments of the beta-limit dextrin well below 30 kDa still show considerable iodine binding. As this extensive degradation is required to achieve the color end-point, the presence of substrate that is susceptible to beta-amylase is likely to influence the final results. 80
M w kDa 800 600 400 200 0 0.4 0 2 4 6 8 10 12 Reaction Time (min) molecular weight iodine binding Figure 5. Change in the weight average molecular weight (Mw) and iodine binding of beta-limit dextrin when incubated with purified malt alpha-amylase under conditions of the ASBC manual method for alpha-amylase. The second portion of the study focused on study of the reaction of beta-limit dextrin with purified malt alpha- and beta-amylase. Malt alpha-amylase was purified from a crude Sigma (Sigma-Aldrich, St Louis) enzyme preparation by heat shock and ultrafiltration. The freeze-dried enzyme preparation was standardized to yield approximately 30, 60, and 90 DU in the manual method. Beta-amylase was from Megazyme (Ireland). In the first series of experiments the loss of iodine binding with alpha-amylase was monitored both in the presence and absence of beta-amylase. 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 OD 650 81
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Annual Pro
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-i- INTRODUCTION The March 2007 <st
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USDA-ARS NORTHERN CROP SCIENCE LABO
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Spring Regional over two years at D
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Table 1. 2006 UCD Two-Rowed Malting
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environments was practiced, 2) elit
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Table 4. Selected six-rowed lines c
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Other Barley Research: Feed Program
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Executive Summary AMBA Prog
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Table 1. Agronomic performance of M
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Lines M122 and M124 were rated sati
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EARLY GENERATION LINES Forty F5 pop
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have evaluated a PCR based marker,
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Edward Schiefelbein, Research Scien
Page 29 and 30: small grains pathology input to thi
Page 31 and 32: Project Personnel Small Grains Path
Page 33 and 34: Investigations on Barley Diseases a
Page 35 and 36: Minnesota barley disease survey and
Page 37 and 38: pathotypes at the seedling stage in
Page 39 and 40: germplasm with the highest level of
Page 41 and 42: Mirlohi, A., Brueggeman, R., Steffe
Page 43 and 44: Developing Improved Malt Barley Var
Page 45 and 46: Table 2. Montana’s Statewide Tria
Page 47 and 48: As a Barley CAP collaborator, our C
Page 49 and 50: 3.2. A barley nursery composed of d
Page 51 and 52: 5. Related Barley Projects Barley P
Page 53 and 54: Barley Breeding - Advanced Testing
Page 55 and 56: Table 3. Agronomic comparisons of N
Page 57 and 58: Preliminary Malting Barley Yield Tr
Page 59 and 60: dwarf genotypes with the convention
Page 61 and 62: STUDIES ON BARLEY DISEASES AND THEI
Page 63 and 64: inoculated plots. The inoculum used
Page 65 and 66: much of North Dakota in 2006, few f
Page 67 and 68: 5. Collect isolates of barley patho
Page 69 and 70: oot rot. In collaboration with Scot
Page 71 and 72: 2. United States Wheat and Barley S
Page 73 and 74: 6. Lee, S. and Neate, S.M. (2006) P
Page 75 and 76: Objectives Met in the One-Year Fund
Page 77 and 78: These automated methods have been t
Page 79: Beta-limit dextrin Alpha-Amylase Al
Page 83 and 84: Figure 7. Time course of I2-binding
Page 85 and 86: 10 9 8 7 6 5 4 3 2 1 0 1993 1994 19
Page 87 and 88: 2. Other Research (A) Predicting Se
Page 89 and 90: forms (through enzymolysis during m
Page 91 and 92: PUBLICATIONS (2006-07) Schwarz, P.B
Page 93 and 94: CI9214, PI552963 (Heartland), and N
Page 95 and 96: showing good levels of resistance t
Page 97 and 98: Germplasm Enhancement for RWA Resis
Page 99 and 100: Gary Hein in Colorado and Nebraska.
Page 101 and 102: Kevin Hicks, Research Leader, USDA-
Page 103 and 104: Detailed Report on
Page 105 and 106: Other Barley Research and Future Di
Page 107 and 108: Table 1. 2006/2007 Oregon winter ba
Page 109 and 110: Table 4. Agronomic data for OSU win
Page 111 and 112: MALTING QUALITY ANALYSIS OF NEW BAR
Page 113 and 114: We recently secured an industrial c
Page 115 and 116: 2006-2007 Progress
Page 117 and 118: enzyme it encodes. We found that se
Page 119 and 120: horseradish peroxidase-conjugated g
Page 121 and 122: found in plant cell walls. The degr
Page 123 and 124: Establishing that sequence variatio
Page 125: Charles Karpelenia, technician Joe
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Annual Progress Report on Malting Barley Research March, 2007

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