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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Crop Improvement and Genetics Research » Research » Publications at this Location » Publication #335365
Title: Improved method for reliable HMW-GS identification by RP-HPLC and SDS-PAGE in common wheat cultivars

Author
item JANG, YOU-RAN - National Academy Of Agricultural Science
item BEOM, HYE-RANG - National Academy Of Agricultural Science
item Altenbach, Susan
item LEE, MIN-KI - National Academy Of Agricultural Science
item LIM, SUN-HYUNG - National Academy Of Agricultural Science
item LEE, JONG-YEOL - National Academy Of Agricultural Science

Submitted to: Molecules
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/22/2017
Publication Date: 6/24/2017
Citation: Jang, Y., Beom, H., Altenbach, S.B., Lee, M., Lim, S., Lee, J. 2017. Improved method for reliable HMW-GS identification by RP-HPLC and SDS-PAGE in common wheat cultivars. Molecules. 22(7):1055. doi:10.3390/molecules22071055.

Interpretive Summary: The high molecular weight glutenin subunits (HMW-GS) are gluten proteins in wheat flour that play an important role in determining the unique functional properties of the flour. The genes encoding these proteins are important targets for wheat breeding efforts to improve flour quality. Because certain HMW-GS are associated with superior quality, it is important to know the exact HMW-GS compositions of wheat lines that will be used in breeding programs. However, there is considerable variability among HMW-GS expressed in different wheat cultivars and small differences in their sequences make these proteins difficult to distinguish. In this manuscript, an improved RP-HPLC method was devised for the separation of HMW-GS that combined with conventional SDS-PAGE made it possible to distinguish HMW-GS variants. The method was used to verify the HMW-GS compositions of 32 Korean wheat cultivars previously determined using SDS-PAGE and to assess the compositions of six new Korean cultivars. The improved RP-HPLC method combined with conventional SDS-PAGE provides for efficient, accurate and cost-effective identification of HMW-GS and will contribute to efforts to improve wheat end-use quality.

Technical Abstract: The accurate identification of alleles for high-molecular weight glutenins (HMW-GS) is critical for wheat breeding programs targeting end-use quality. RP-HPLC methods were optimized for separation of HMW-GS, resulting in enhanced resolution of 1By and 1Dx subunits. Statistically significant differences in retention times (RTs) for subunits corresponding to HMW-GS alleles were determined using 16 standard wheat cultivars with known HMW-GS compositions. Subunits that were not identified unambiguously by RP-HPLC were distinguished by SDS-PAGE or inferred from association with linked subunits. The method was used to verify the allelic compositions of 32 Korean wheat cultivars previously determined using SDS-PAGE and to assess the compositions of six new Korean cultivars. Three cultivars contained subunits that were identified incorrectly in the earlier analysis. The improved RP-HPLC method combined with conventional SDS-PAGE provides for efficient, accurate and cost-effective identification of HMW-GS and will contribute to efforts to improve wheat end-use quality.