Annual Progress Report on Malting Barley Research March, 2002

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Table 1. Results of the 2001 barley disease survey (11 fields visited) in Minnesota. Disease/Pathogen Fusarium head blight (Fusarium spp.) Net blotch (Pyrenophora teres f. teres) Wheat stem rust (Puccinia graminis f. sp. tritici) Septoria/Stagonospora speckled leaf blotch (Septoria and Stagonospora spp.) Loose smut (incidence) (Ustilago nuda) Crown rust (Puccinia coronata var. hordei) Barley yellow dwarf (Barley yellow dwarf virus BYDV) Bacterial blight (Xanthomonas translucens pv. Translucens) 50 Number of fields with disease Disease severity or incidence 11 Trace to 4% 11 Trace to 80% 8 Trace to 3% 7 Trace to 5% 7 Trace 8 Trace to 15% 4 100% 1 20%
51 BARLEY TRANSFORMATION G.J. Muehlbauer, C.A. Mackintosh, D.T. Tuong and N. Al-Saady Department of Agronomy and Plant Genetics, University of Minnesota St. Paul, MN 55108 Summary 1. We have modified a protocol for transforming the elite barley variety Conlon in our laboratory. 1500, 1105 and 40 callus pieces have been bombarded with gold particles coated with DNA encoding the following antifungal proteins (AFP): a wheat thaumatinlike protein 1 (tlp-1), a barley chitinase and a barley glucanase. Many regenerating plantlets are in the final stages of tissue culture. Further bombardments using the barley glucanase and other AFP genes are on-going. 2. We have obtained affinity purified polyclonal antibodies to the following proteins; a barley ribosome inactivating protein (RIP), a wheat tlp-1, a barley chitinase and a barley glucanase. Protocols for using these antibodies in Western blotting to determine AFP expression are being developed using transgenic wheat. Thus, we will be able to determine the presence of these proteins in our transgenic barley as soon as the plants are growing in soil. 3. We are developing in vitro fungal growth assays in order to identify antifungal properties of transgenic barley seedlings. This will allow us to determine potential antifungal effects before the plants reach flowering, the growth stage at which Fusarium graminearum-glasshouse disease screening occurs. 4. We have continued to characterize barley plants transformed with the GUS gene driven by the sugarcane badnavirus (ScBV) promoter. As described in the AMBA 2000- 2001 progress report, we developed twelve transgenic lines of barley expressing GUS driven by the ScBV promoter. In the T1 generation we showed that the ScBV promoter drives expression in all barley tissues in all twelve transgenic lines. Our work within the last year was directed at determining the stability of GUS expression over generations. Therefore, we have evaluated T2 generation lines. In the T2 generation, GUS was expressed in eight of the 12 transgenic lines and appeared to be silenced in the other four lines. Introduction Genetic transformation promises to be a powerful tool for improvement of crop plants. Genetic manipulations that are not possible using conventional breeding methods are attainable with plant transformation. Barley transformation has been accomplished via particle bombardment into immature zygotic embryos, callus-derived immature embryos and microspore-derived embryos (Wan and Lemaux, 1994). Fusarium head blight (FHB), a fungal disease of small grain crops caused by Fusarium graminearum, threatens to reduce barley to an economically unviable crop in Minnesota and many other states. Currently, large breeding efforts at the University of Minnesota and around the country are focused on developing FHB resistant cultivars. However,
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Annual Pro
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-i- INTRODUCTION The March 2002 <st
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Barley Transformation G.J. Muehlbau
Page 9 and 10: 1 2000/2001 WINTER NURSERY: BARLEY
Page 11 and 12: 3 2001 Barley Stripe Rust Screening
Page 13 and 14: 5 Triumph/Tyra//Arupo, a selection
Page 15 and 16: 7 Washington (Fossum Cereals and Wa
Page 17 and 18: 9 This project is supported by the
Page 19 and 20: 11 ‘Triumph’, ‘Valier’, ‘
Page 21 and 22: 13 Table 2. Agronomic data for sele
Page 23 and 24: 15 Table 4. Agronomic data for sele
Page 25 and 26: 17 Table 8. Malting quality data* f
Page 27 and 28: 19 Six-rowed barley selections of c
Page 29 and 30: 21 Table 14. Agronomic data for sel
Page 31 and 32: 23 Table 16. Agronomic data for win
Page 33 and 34: 25 Table 19. Malting quality data*
Page 35 and 36: Garnet/98Ab11865 Garnet/98Ab12895 9
Page 37 and 38: 29 rust resistant NSGC accessions o
Page 39 and 40: 31 DEVELOPMENT OF SIX-ROWED MALTING
Page 41 and 42: 33 backcrossing. The selfed lines w
Page 43 and 44: 35 MINNESOTA BARLEY IMPROVEMENT PRO
Page 45 and 46: 37 ADVANCED LINE EVALUATION Varieti
Page 47 and 48: 39 OTHER BARLEY DISEASES In 2001, r
Page 49 and 50: 41 Wingbermuehle, W. J., K.M. Belin
Page 51 and 52: 43 In 2001 winter/spring, over 300
Page 53 and 54: 45 assessments were made by countin
Page 55 and 56: 47 IDENTIFICATION OF NOVEL DISEASE
Page 57: 49 Unfortunately, disease levels in
Page 61 and 62: 53 We have available the sugarcane
Page 63 and 64: 55 Secondly, we tried to grow F. gr
Page 65 and 66: 57 References: Issac, S. and Jennin
Page 67 and 68: 59 Huntley-dryland) with two replic
Page 69 and 70: 61 Table 2. 1992 thru 2001 Overall
Page 71 and 72: 63 performance for Montana farmers.
Page 73 and 74: 65 Blake, T,. Hensleigh P., Boss D.
Page 75 and 76: 67 settling tower cannot accommodat
Page 77 and 78: Table 3. Seedling reaction to strip
Page 79 and 80: 71 TWO-ROWED BARLEY IMPROVEMENT PRO
Page 81 and 82: Table 1. Agronomic trait comparison
Page 83 and 84: 75 mutants reduce plant vigor and g
Page 85 and 86: 77 SIX-ROWED BARLEY IMPROVEMENT PRO
Page 87 and 88: 79 this variety has decreased over
Page 89 and 90: Table 7. Malt quality comparisons o
Page 91 and 92: 83 • Management studies for malt
Page 93 and 94: 85 STUDIES ON BARLEY DISEASES AND T
Page 95 and 96: 87 With funding from the US Wheat a
Page 97 and 98: 89 MALTING AND BREWING QUALITY OF B
Page 99 and 100: 91 β-glucans, it might be most ins
Page 101 and 102: 10 9 8 7 6 5 4 3 2 1 0 93 Figure 1.
Page 103 and 104: 95 Bolin, P., Schwarz, P., Jones, B
Page 105 and 106: 97 Results Results from field and g
Page 107 and 108: 99 DEVELOPING BARLEY TISSUE CULTURE
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101 trials of plants regenerated fr
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103 Table 2. Agronomic performance
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105 BSMV spread between entries. Ho
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107 involving two susceptible backg
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109 THE OREGON BARLEY IMPROVEMENT P
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111 for traits that are difficult/e
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Germplasm Development: 113 Crosses:
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115 • All lines in spring yield t
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117 Table 2. Agronomic data for Ore
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119 Table 6. Malting quality of win
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121 Table 7. Across location summar
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123 MOLECULAR MARKER ASSISTED MODIF
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125
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127 exchanges. Spring and winter ba
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129 Table 2. 2000-2001 Agronomic Pe
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131 Table 7. Spring 2- and 6-Row #
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133 2. Direct Seeding Cropping Syst
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135 D.M. Wesenberg - spring and win
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137 A STUDY OF THE MALTING QUALITY
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139 Methods. Our malting schedule.
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141 modification measures indicated
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143 CHARACTERIZATION OF THE PROTEIN
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145 Purification of a serine endope
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147 STUDIES ON THE PROTEINASES THAT
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149 with the proteinase T will be s
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151 10°C increase in thermostabili
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153 Sissons MJ and MacGregor AW (19
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µM Carbohydrate / Unit of α-Gluco
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157 Tissue-specific gene products f
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159 The promoters were subcloned fr
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161 Fig. 3. Expression of Trx-Hth f
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Annual Progress Report on Malting Barley Research March, 2002

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