Jang You-Ran, Beom Hye-Rang, Altenbach Susan B, Lee Min-Ki, Lim Sun-Hyung, Lee Jong-Yeol
National Institute of Agricultural Science, RDA, Jeonju 54874, Korea.
USDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, USA.
Molecules. 2017 Jun 24;22(7):1055. doi: 10.3390/molecules22071055.
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 accurate, efficient and reliable identification of HMW-GS and will contribute to efforts to improve wheat end-use quality.
准确鉴定高分子量麦谷蛋白（HMW-GS）的等位基因对于以最终用途品质为目标的小麦育种计划至关重要。对反相高效液相色谱（RP-HPLC）方法进行了优化以分离HMW-GS，从而提高了1By和1Dx亚基的分辨率。使用16个已知HMW-GS组成的标准小麦品种，确定了与HMW-GS等位基因相对应的亚基在保留时间（RT）上的统计学显著差异。通过SDS-PAGE区分了RP-HPLC未能明确鉴定的亚基，或从与连锁亚基的关联中推断出来。该方法用于验证先前使用SDS-PAGE确定的32个韩国小麦品种的等位基因组成，并评估6个新韩国品种的组成。三个品种含有在早期分析中鉴定错误的亚基。改进后的RP-HPLC方法与传统的SDS-PAGE相结合，可准确、高效且可靠地鉴定HMW-GS，并将有助于提高小麦最终用途品质的努力。
