Location: Crop Improvement and Genetics ResearchTitle: Proteomic determination of low-molecular-weight glutenin subunit composition in Aroona near-isogenic lines and standard wheat cultivars
|CHO, KYOUNGWON - National Institute For Agricultural Science & Technology
|JANG, YOU-RAN - Rural Development Administration - Korea
|LIM, SUN-HYUNG - Hankyong National University
|SIMON-BUSS, ANNAMARIA - University Of Hamburg
|LEE, JONG-YEOL - National Institute For Agricultural Science & Technology
Submitted to: International Journal of Molecular Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/15/2021
Publication Date: 7/19/2021
Citation: Cho, K., Jang, Y., Lim, S., Altenbach, S.B., Gu, Y.Q., Simon-Buss, A., Lee, J. 2021. Proteomic determination of low-molecular-weight glutenin subunit composition in Aroona near-isogenic lines and standard wheat cultivars. International Journal of Molecular Sciences. 22(14). Article: 7709. https://doi.org/10.3390/ijms22147709.
Interpretive Summary: Low-molecular weight glutenin subunits (LMW-GS) are a complex group of proteins that link with high-molecular weight glutenin subunits (HMW-GS) to form large protein polymers that are essential for determining the viscoelasticity properties of wheat flour. The contributions of individual LMW-GS proteins are difficult to access because of the complex composition in individual wheat cultivars as well as variability of compositions among different wheat cultivars. This study uses reversed-phase HPLC and 2-dimensional gel electrophoresis combined with tandem mass spectrometry to discriminate individual LMW-GS proteins. Knowledge about LMW-GS composition is essential for selecting genes that will result in wheat with improved flour quality.
Technical Abstract: The low-molecular weight glutenin subunit (LMW-GS) composition of wheat (Triticum aestivum) flour has important effects on end-use quality. However, assessing the contributions of each LMW-GS to flour quality remains challenging because of the complex LMW-GS composition and allelic variation among wheat cultivars. Therefore, accurate and reliable determination of LMW-GS alleles in germplasm remains an important challenge for wheat breeding. In this study, we used an optimized reversed-phase HPLC method and proteomics approach comprising 2-D gels coupled with liquid chromatography-tandem mass spectrometry (MS/MS) to discriminate individual LMW-GSs corresponding to alleles encoded by the Glu-A3, Glu-B3, and Glu-D3 loci in the ‘Aroona’ cultivar and 12 ‘Aroona’ near-isogenic lines (ARILs), which contain unique LMW-GS alleles in the same genetic background. The LMW-GS separation patterns for ‘Aroona’ and ARILs on chromatograms and 2-D gels were consistent with those from a set of 10 standard wheat cultivars for Glu-3. Furthermore, 12 previously uncharacterized spots in ‘Aroona’ and ARILs were excised from 2-D gels, digested with chymotrypsin, and subjected to MS/MS. We identified their gene haplotypes and created a 2-D gel map of LMW-GS alleles in the germplasm for breeding and screening for desirable LMW-GS alleles for wheat quality improvement.