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 (<i>Triticum aestivum</i>) 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 alleli...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7709 |
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| author | Kyoungwon Cho You-Ran Jang Sun-Hyung Lim Susan B. Altenbach Yong Q. Gu Annamaria Simon-Buss Jong-Yeol Lee |
| author_facet | Kyoungwon Cho You-Ran Jang Sun-Hyung Lim Susan B. Altenbach Yong Q. Gu Annamaria Simon-Buss Jong-Yeol Lee |
| author_sort | Kyoungwon Cho |
| collection | DOAJ |
| description | The low-molecular weight glutenin subunit (LMW-GS) composition of wheat (<i>Triticum aestivum</i>) 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 <i>Glu-A3</i>, <i>Glu-B3</i>, and <i>Glu-D3</i> 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 <i>Glu-3</i>. 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. |
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| language | English |
| last_indexed | 2024-03-10T09:36:19Z |
| publishDate | 2021-07-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6ebf82111b69495fb71e7be8ae6140fa2023-11-22T04:02:57ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214770910.3390/ijms22147709Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat CultivarsKyoungwon Cho0You-Ran Jang1Sun-Hyung Lim2Susan B. Altenbach3Yong Q. Gu4Annamaria Simon-Buss5Jong-Yeol Lee6Department of Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61186, KoreaNational Institute of Agricultural Science, RDA, Jeonju 54874, KoreaDivision of Horticultural Biotechnology, Hankyong National University, Anseong 17579, KoreaUSDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, USAUSDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, USAUSDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, USANational Institute of Agricultural Science, RDA, Jeonju 54874, KoreaThe low-molecular weight glutenin subunit (LMW-GS) composition of wheat (<i>Triticum aestivum</i>) 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 <i>Glu-A3</i>, <i>Glu-B3</i>, and <i>Glu-D3</i> 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 <i>Glu-3</i>. 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.https://www.mdpi.com/1422-0067/22/14/7709LMW-GSproteomic analysisRP-HPLCAroona near-isogenic linesstandard wheat cultivars |
| spellingShingle | Kyoungwon Cho You-Ran Jang Sun-Hyung Lim Susan B. Altenbach Yong Q. Gu Annamaria Simon-Buss Jong-Yeol Lee Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars International Journal of Molecular Sciences LMW-GS proteomic analysis RP-HPLC Aroona near-isogenic lines standard wheat cultivars |
| title | Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars |
| title_full | Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars |
| title_fullStr | Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars |
| title_full_unstemmed | Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars |
| title_short | Proteomic Determination of Low-Molecular-Weight Glutenin Subunit Composition in Aroona Near-Isogenic Lines and Standard Wheat Cultivars |
| title_sort | proteomic determination of low molecular weight glutenin subunit composition in aroona near isogenic lines and standard wheat cultivars |
| topic | LMW-GS proteomic analysis RP-HPLC Aroona near-isogenic lines standard wheat cultivars |
| url | https://www.mdpi.com/1422-0067/22/14/7709 |
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