Annual Progress Report on Malting Barley Research March, 2002

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7 6 5 4 3 2 1 0 56 Figure 1: Weight of Fusarium graminearum mycelium grown in liquid culture. Results are the means ±SEM of duplicate measurements. * indicates only one measurement made Weight of Fusarium graminearum mycelium (g) Control * 10 mM EDTA 1 mM EDTA 100 µM EDTA Treatment 10 µM EDTA * 1 µM EDTA 100 nM EDTA 4. CHARACTERIZATION OF THE SCBV PROMOTER From twelve T0 events, 5-10 T1 seeds were planted to produce T1 plants. These T1 plants were analyzed for GUS expression. GUS was expressed highly in the roots, stems, leaves and all reproductive tissues of the barley plants, the ovaries, anthers, bracts and rachis. T2 seed from each of those twelve lines was then planted out and the T2 plants analyzed for GUS expression. In eight of the lines, the expression remained constant, as it had in the previous generation. However, in four of the lines, GUS expression was silenced. Nevertheless, these data indicate that the ScBV promoter will be a useful promoter for transforming barley. Future work - 1 st March 2002 - 30 th June 2002 Conlon stock plants are planted every two weeks. Therefore, an almost continuous supply of explant material is available. In the remaining four months of this project, we will increase the number of callus pieces bombarded with our barley glucanase gene from 40 to approximately 1000. We will also bombard approximately 1000 callus pieces with our barley RIP gene. The success rate of barley transformation is approximately 1%, thus, 1000 callus pieces are necessary to produce a significant number of transgenic events. We will begin to characterize transgenic plants that are presently in the late stages of tissue culture, utilizing our antibodies. We will begin to advance them to the next generation. We will also test transgenics that express AFPs for in vitro antifungal effects.
57 References: Issac, S. and Jennings, D.H. 1995. Microbial culture. Bios Scientific Publishing, Oxford. Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusion: ß-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6:3901- 3907. Tingay, S., McElroy, D., Kalla, R., Fieg, S., Wang, M., Thornton, S. and Brettell, R. 1997. Agrobacterium tumefaciens-mediated barley transformation. Plant J. 11:1369- 1376. Tzafrir, I., Torbert, K.A., Lockhart, B.E.L., Somers, D.A. and Olszewski N.E. 1998. The sugarcane bacilliform badnavirus promoter is active in both monocots and dicots. Plant Mol. Biol. 38:347-356. Wan, Y. and Lemaux, P.G. 1994. Generation of large numbers of independently transformed fertile barley plants. Plant Physiol. 104:37-48. Personnel: Caroline A. Mackintosh, Postdoctoral Research Associate Dong T. Tuong, Technician Nadiya Al-Saady, Ph.D. student Collaborators: Dr. Ann Blechl, USDA-ARS, Albany, CA Dr. David Somers, University of Minnesota Dr. Richard Zeyen, University of Minnesota
Page 1 and 2:
Annual Pro
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-i- INTRODUCTION The March 2002 <st
Page 6 and 7:
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 and 58: 49 Unfortunately, disease levels in
Page 59 and 60: 51 BARLEY TRANSFORMATION G.J. Muehl
Page 61 and 62: 53 We have available the sugarcane
Page 63: 55 Secondly, we tried to grow F. gr
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
Page 109 and 110: 101 trials of plants regenerated fr
Page 111 and 112: 103 Table 2. Agronomic performance
Page 113 and 114: 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
Page 127 and 128:
119 Table 6. Malting quality of win
Page 129 and 130:
121 Table 7. Across location summar
Page 131 and 132:
123 MOLECULAR MARKER ASSISTED MODIF
Page 133 and 134:
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
Page 145 and 146:
137 A STUDY OF THE MALTING QUALITY
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139 Methods. Our malting schedule.
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141 modification measures indicated
Page 151 and 152:
143 CHARACTERIZATION OF THE PROTEIN
Page 153 and 154:
145 Purification of a serine endope
Page 155 and 156:
147 STUDIES ON THE PROTEINASES THAT
Page 157 and 158:
149 with the proteinase T will be s
Page 159 and 160:
151 10°C increase in thermostabili
Page 161 and 162:
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
Page 167 and 168:
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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