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Changes in High Molecular Weight Glutenin Subunit Composition Can Be Genetically Engineered without Affecting Wheat Agronomic Performance

^a USDA-ARS National Small Grains Germplasm Research Facility, 1691 S. 2700 W., Aberdeen, ID 83211
^b USDA-ARS Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710
^c Dep. of Plant Sciences, University of California, Davis, CA 95616
^d USDA, ARS, KSARC, 2413 E. Hwy 83, Weslaco, TX 78596
^e Desert Research and Extension Center, University of California, El Centro, CA 92243

View larger version (15K): [in a new window]   Fig. 1. Diagrammatic representation of the high molecular weight glutenin subunit (HMW-GS) DNA sequences introduced by genetic transformation. Thickest boxes are coding regions, medium boxes are 5' flanking regions including promoters, and thin boxes are 3' flanking regions including transcription termination and polyA addition sites. Sequences from the native wheat gene Glu-D1–2 (Dy10) are in black, native gene GluD1–1 (Dx5) are in white, and native gene Glu-A1–1 (Ax2*) are in diagonal stripes. The coding sequences are divided into three regions corresponding to HMW-GS protein domains: the short N-terminal nonrepetitive region, the long repetitive region, and the short C-terminal nonrepetitive region shared by all HMW-GS. Shorthand names used in this report for the HMW-GS are to the left. (The pBluescript plasmid backbones are not shown.)

View larger version (114K): [in a new window]   Fig. 2. SDS-PAGE of seed proteins in transgenic wheat lines (numbered lanes) and their nontransgenic parent Bobwhite (C). Arrowheads to the left of the individual lanes indicate expected locations of bands corresponding to transgene-encoded high molecular weight glutenin subunit (HMW-GS). The gel positions and names of the HMW-GS native to Bobwhite are indicated to the right. (A) Numbered lanes contain protein extracts from greenhouse-grown seeds from homozygous transgenic wheat lines expressing transgene-encoded native HMW-GS: T3 Ax2*-A (1), T3 Dy10-A (2), T6 Dy10-E (3), T5 Dx5-G (4), T5 Dx5-J (5), T7 Dx5+Dy10-A (6), T7 Dx5+Dy10-D (7), and T7 Dx5+Dy10-G (8). (B) Numbered lanes contain protein extracts from greenhouse-grown seeds from homozygous transgenic wheat lines expressing transgene-encoded recombinant HMW-GS: T11 Hybrid-C (1), T12 Hybrid-G (2), T6 LongDx5-C (3), T5 LongDx5-G (4), T4 ShortDx5-A (5), and T8 ShortDx5-D (6). (C) Numbered lanes contain protein extracts from greenhouse-grown—except Hybrid-B, which was field-grown—seeds from homozygous transgenic wheat lines exhibiting transgene-mediated cosuppression of HMW-GS: T9 Hybrid-A (1), T5 LongDx5-A (2), T4 Dx5-D (3), T5 Dx5-A (4), and T7 Hybrid-B (5). Lane BAR contains T11 seed extract from BAR-D.

View larger version (93K): [in a new window]   Fig. 3. SDS-PAGE of seed proteins in the lower-yielding transgenic wheat lines (numbered lines) and their nontransgenic parent Bobwhite (C). The gel positions and names of the high molecular weight glutenin subunit (HMW-GS) native to Bobwhite are indicated to the left. Numbered lanes contain protein extracts from field-grown seeds from homozygous transgenic wheat lines T11 Hybrid-C (1), T7 Hybrid-B (2), T6 Dx5 + Dy10-E (3), T5 Dx5-L (4), T5 ShortDx5-F (5), T8 LongDx5-E short (6), and T5 Dx5 + Dy10-C (7). Arrowheads to the right of the individual lanes indicate expected locations of bands corresponding to transgene-encoded HMW-GS. Open circles to the right of individual lanes mark locations of bands of unexpected sizes and unknown identity.