Rapid Identification of HMW Glutenin Subunits from Different Hexaploid Wheat Species by Acidic Capillary Electrophoresis

Abstract

Cereal Chem. 81(5):561-566High molecular weight glutenin subunits (HMW-GS) from three hexaploid wheat species (AABBDD, 2n=6x=42, Triticum aestivum L., T. spelta L., and T. compactum L.) were separated and identified by acidic capillary electrophoresis (A-CE) with phosphate-glycine buffer (pH 2.5) in uncoated fused-silica capillaries (50 µm, i.d. × 25.5 cm) at 12.5 kV and 40°C. The rapid separations (<15 min) of HMW-GS with good repeatability (RSD < 2%) were obtained using a fast capillary rising protocol. All 17 HMW-GS analyzed could be well separated and their relative migration orders were ranked. In particular, the good quality subunit pair 5+10 could be differentiated from poor quality subunit pair 2+12. In addition, the other three allelic pairs of 13+16, 17+18, and 7+8 subunits that were considered to have positive effects on dough properties, as well as three pairs of novel subunits 13+22

Characterization and Comparative Analysis of Wheat High Molecular Weight Glutenin Subunits by SDS-PAGE, RP-HPLC, HPCE, and MALDI-TOF-MS

“…The HMW-GS elution order under acidic buffer CE condition was 1Dy f 1By f 1Bx f 1Ax f 1Dx. As indicated by previous investigations (13,14,35), the peaks of individual subunits displayed isoforms under acidic separation condition and similar results were obtained by two-dimensional gel electrophoresis (data not shown), which probably resulted from sample treatment or post-translational modifications (PTMs). In most cases, each subunit had one major peak and one or two minor peaks.…”

“…Therefore, HMW-GS have become useful protein markers for wheat quality improvement and cultivar identification. In the past several decades, considerable efforts have been focused on developing powerful techniques for fast differentiation of good-and poor-quality HMW-GS in wheat for germplasm screening and quality improvement (11)(12)(13)(14)(15). Up to now, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) has been the most popular method to detect the allelic compositions of HMW-GS on the basis of their mobilities (16).…”

Capillary electrophoresis of proteins 2003–2005

“…Analysis of proteins in cereal cultivars has widely shown the importance of CE of cereal proteins for quality control [220][221][222][223][224][225][226][227][228]. Alcohol extracts from wheat grain typically contain gliadins, whereas water extracts contain mainly albumins.…”

“…For example, CZE has been used for identification of glutenin subunits from different hexaploid wheat species [222,226]. Lines belonging to the same species can be discriminated mainly on the basis of b-and o-glutenin patterns, while g-and ogliadins seem to be more suitable for differentiation of emmer (durum wheat) from spelt (common wheat) [229].…”

Rapid separation and characterization of grain water-soluble proteins in bread wheat cultivars (Triticum aestivum L.) by capillary electrophoresis

“…For the characterization of high molecular weight glutenin subunits of wheat, Yan et al (2004) developed a rapid method of acidic capillary electrophoresis. In the present study, we focused on developing a rapid CE method for the separation and characterization of seed WS proteins of bread wheats.…”