Characterization of HMW Glutenin Subunits in Bread and Tetraploid Wheats by Reversed-Phase High-Performance Liquid Chromatography
The wheat storage proteins, especially the high molecular weight glutenin subunits (HMW-GS), play important roles in the determination of flour processing and bread-making quality. Compared with the traditional SDS-PAGE method, reversed-phase high-performance liquid chromatography (RP-HPLC) was shown to have many advantages for the separation and characterization of HMW-GS because of its high resolving power, repeatability and automation. In this work, HMW-GS from bread and tetraploid wheats were separated and characterized by RP-HPLC. The elution time ranking of different HMW-GS was: 1Ax > 1Bx > 1Dx > 1By > 1Dy. Several subunit pairs associated with good quality properties and those with similar mobilities on SDS-PAGE, such as 1Bx7 and 1Bx7*, 1By8 and 1By8*, 1Dx2 and 1Ax2*, 1Bx6 and 1Bx6.1, were well separated and readily identified through RP-HPLC. However, other subunit pairs, such as 1Dy10 — 1Dy12, 1Dx5 — 1By18 and 1Dx2 — 1By16, could not be adequately separated and identified by RP-HPLC, whereas they displayed different mobilities on SDS-PAGE gels. Because 1Dx5 and 1Dx2 showed different hydrophobicities, RP-HPLC could distinguish 1Dx5 + 1Dy10 and 1Dx2 + 1Dy12. A comparative analysis between RP-HPLC and SDS-PAGE showed that a combination of both methods provided more effective identification of HMW-GS in wheat quality improvement and germplasm screening.
Keywordswheat HMW-GS RP-HPLC
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