Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars

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 w...

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Bibliographic Details
Published in:International journal of molecular sciences 2021-07, Vol.22 (14), p.7709
Main Authors: Cho, Kyoungwon, Jang, You-Ran, Lim, Sun-Hyung, Altenbach, Susan B., Gu, Yong Q., Simon-Buss, Annamaria, Lee, Jong-Yeol
Format: Article
Language:English
Subjects:
Alleles
Aroona near-isogenic lines
Chromosomes
Chymotrypsin
Composition
Cultivars
Flour
Gels
Genes
Germplasm
Glutenin
Haplotypes
High performance liquid chromatography
LMW-GS
Mass spectrometry
Mass spectroscopy
Molecular weight
Plant breeding
proteomic analysis
Proteomics
Quality assessment
Quality control
Retention
RP-HPLC
standard wheat cultivars
Subunit structure
Triticum aestivum
Wheat
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Summary: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.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms22147709