Abstract
Lab-on-a-chip technology is an emerging and convenient system to easily and quickly separate proteins of high molecular weight. The current study established a high-molecular-weight glutenin subunit (HMW-GS) identification system using Lab-on-a-chip for three, six, and three of the allelic variations at the Glu-A1, Glu-B1, and Glu-D1 loci, respectively, which are commonly used in wheat breeding programs. The molecular weight of 1Ax1 and 1Ax2* encoded by Glu-A1 locus were of 200 kDa and 192 kDa and positioned below 1Dx subunits. The HMW-GS encoded by Glu-B1 locus were electrophoresed in the following order below 1Ax1 and 1Ax2*: 1Bx13 ≥ 1Bx7 = 1Bx7OE > 1Bx17 > 1By16 > 1By8 = 1By18 > 1By9. 1Dx2 and Dx5 showed around 4-kDa difference in their molecular weights, with 1Dy10 and 1Dy12 having 11-kDa difference, and were clearly differentiated on Lab-on-a-chip. Additionally, some of the HMW-GS, including 1By8, 1By18, and 1Dy10, having different theoretical molecular weights showed similar electrophoretic mobility patterns on Lab-on-a-chip. The relative protein amount of 1Bx7OE was two-fold higher than that of 1Bx7 or 1Dx5 and, therefore, translated a significant increase in the protein amount in 1Bx7OE. Similarly, the relative protein amounts of 8 & 10 and 10 & 18 were higher than each subunit taken alone. Therefore, this study suggests the established HMW-GS identification system using Lab-on-a-chip as a reliable approach for evaluating HMW-GS for wheat breeding programs.
Highlights
Wheat (Triticum aestivum L.) is an important staple food crop, which provides substantial amounts of various components, such as proteins and vitamins, that are essential for human consumption and health and the industry
Compositions using standard cultivars (Table 1). These cultivars covered three, six, and three of the allelic variations at the Glu-A1, Glu-B1, and Glu-D1 loci, respectively, which are commonly used in wheat breeding programs
The recorded molecular weights of high-molecular-weight glutenin subunit (HMW-GS) on Lab-on-a-chip were above 120 kDa [31]
Summary
Wheat (Triticum aestivum L.) is an important staple food crop, which provides substantial amounts of various components, such as proteins and vitamins, that are essential for human consumption and health and the industry. In addition to being an important source of energy, wheat serves as an ingredient for diverse foods due to the presence of the seed storage protein gluten [1], which is built up of subunits and imparts elasticity to a dough [2]. An x-type and a y-type subunits of HMW-GS encoded in each Glu-1 locus are located on the long arms of chromosome 1 on the A, B, and D genomes of bread wheat [5]. These genes on the Glu-1 loci are tightly linked, considering that the physical distance between an x-type and a y-type subunits ranges
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