Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography
Wheat (<i>Triticum aestivum</i> L.) omega-5 gliadin, a major allergen responsible for wheat-dependent exercise-induced anaphylaxis in humans, is encoded by genes located at the <i>Gli-B1</i> locus on chromosome 1B, which exhibits genetic polymorphism. <i>Gli-B1</i>...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-01-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/30/3/609 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850068446072537088 |
|---|---|
| author | Jong-Yeol Lee Yu-Jeong Yang Jinpyo So Sewon Kim Kyoungwon Cho |
| author_facet | Jong-Yeol Lee Yu-Jeong Yang Jinpyo So Sewon Kim Kyoungwon Cho |
| author_sort | Jong-Yeol Lee |
| collection | DOAJ |
| description | Wheat (<i>Triticum aestivum</i> L.) omega-5 gliadin, a major allergen responsible for wheat-dependent exercise-induced anaphylaxis in humans, is encoded by genes located at the <i>Gli-B1</i> locus on chromosome 1B, which exhibits genetic polymorphism. <i>Gli-B1</i> alleles have generally been identified based on the electrophoretic mobilities of the encoded gamma-, omega-1,2, and omega-5 gliadins in acid polyacrylamide gel electrophoresis. However, the similar mobilities of omega-5 gliadin variants make it difficult to distinguish them among different wheat varieties. In this study, we optimized reverse-phase–ultra-performance liquid chromatography (RP-UPLC) conditions to separate omega-5 gliadins in the reference wheat cultivar Chinese Spring and its nullisomic–tetrasomic lines for chromosome 1B. Five chromatographic peaks corresponded to omega-5 gliadin, and the average relative standard deviation to each peak retention time ranged from 0.31% to 0.93%, indicating that the method is accurate and reproducible for fractionating omega-5 gliadins in gliadin extracts from wheat flour. Using the optimized RP-UPLC method, we analyzed omega-5 gliadins in 24 wheat varieties with the <i>Gli-B1f</i> allele. The result showed that the wheat varieties were sorted into eight groups according to the composition of omega-5 gliadin, indicating that the classification of <i>Gli-B1</i> alleles based on A-PAGE could not explain the composition of omega-5 gliadin in wheat. We reclassified 73 wheat varieties containing 16 unique <i>Gli-B1</i> alleles into 31 groups based on the chromatographic patterns of their omega-5 gliadins. Our results provide information on the specific <i>Gli-B1</i> alleles of wheat varieties belonging to each group and demonstrate the potential for RP-UPLC to facilitate genetic studies of wheat varieties. |
| format | Article |
| id | doaj-art-cede3f9b86ba4a1b95fc2d79a8e4ed21 |
| institution | DOAJ |
| issn | 1420-3049 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-cede3f9b86ba4a1b95fc2d79a8e4ed212025-08-20T02:48:03ZengMDPI AGMolecules1420-30492025-01-0130360910.3390/molecules30030609Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid ChromatographyJong-Yeol Lee0Yu-Jeong Yang1Jinpyo So2Sewon Kim3Kyoungwon Cho4National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, Republic of KoreaNational Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, Republic of KoreaDepartment of Integrative Food, Bioscience and Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61166, Republic of KoreaNational Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, Republic of KoreaDepartment of Integrative Food, Bioscience and Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 61166, Republic of KoreaWheat (<i>Triticum aestivum</i> L.) omega-5 gliadin, a major allergen responsible for wheat-dependent exercise-induced anaphylaxis in humans, is encoded by genes located at the <i>Gli-B1</i> locus on chromosome 1B, which exhibits genetic polymorphism. <i>Gli-B1</i> alleles have generally been identified based on the electrophoretic mobilities of the encoded gamma-, omega-1,2, and omega-5 gliadins in acid polyacrylamide gel electrophoresis. However, the similar mobilities of omega-5 gliadin variants make it difficult to distinguish them among different wheat varieties. In this study, we optimized reverse-phase–ultra-performance liquid chromatography (RP-UPLC) conditions to separate omega-5 gliadins in the reference wheat cultivar Chinese Spring and its nullisomic–tetrasomic lines for chromosome 1B. Five chromatographic peaks corresponded to omega-5 gliadin, and the average relative standard deviation to each peak retention time ranged from 0.31% to 0.93%, indicating that the method is accurate and reproducible for fractionating omega-5 gliadins in gliadin extracts from wheat flour. Using the optimized RP-UPLC method, we analyzed omega-5 gliadins in 24 wheat varieties with the <i>Gli-B1f</i> allele. The result showed that the wheat varieties were sorted into eight groups according to the composition of omega-5 gliadin, indicating that the classification of <i>Gli-B1</i> alleles based on A-PAGE could not explain the composition of omega-5 gliadin in wheat. We reclassified 73 wheat varieties containing 16 unique <i>Gli-B1</i> alleles into 31 groups based on the chromatographic patterns of their omega-5 gliadins. Our results provide information on the specific <i>Gli-B1</i> alleles of wheat varieties belonging to each group and demonstrate the potential for RP-UPLC to facilitate genetic studies of wheat varieties.https://www.mdpi.com/1420-3049/30/3/609omega-5 gliadinreverse-phase–ultra-performance liquid chromatography (RP-UPLC)<i>Gli-B1</i> locus |
| spellingShingle | Jong-Yeol Lee Yu-Jeong Yang Jinpyo So Sewon Kim Kyoungwon Cho Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography Molecules omega-5 gliadin reverse-phase–ultra-performance liquid chromatography (RP-UPLC) <i>Gli-B1</i> locus |
| title | Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography |
| title_full | Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography |
| title_fullStr | Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography |
| title_full_unstemmed | Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography |
| title_short | Allelic Analysis of the <i>Gli-B1</i> Locus in Hexaploid Wheat Using Reverse-Phase–Ultra-Performance Liquid Chromatography |
| title_sort | allelic analysis of the i gli b1 i locus in hexaploid wheat using reverse phase ultra performance liquid chromatography |
| topic | omega-5 gliadin reverse-phase–ultra-performance liquid chromatography (RP-UPLC) <i>Gli-B1</i> locus |
| url | https://www.mdpi.com/1420-3049/30/3/609 |
| work_keys_str_mv | AT jongyeollee allelicanalysisoftheiglib1ilocusinhexaploidwheatusingreversephaseultraperformanceliquidchromatography AT yujeongyang allelicanalysisoftheiglib1ilocusinhexaploidwheatusingreversephaseultraperformanceliquidchromatography AT jinpyoso allelicanalysisoftheiglib1ilocusinhexaploidwheatusingreversephaseultraperformanceliquidchromatography AT sewonkim allelicanalysisoftheiglib1ilocusinhexaploidwheatusingreversephaseultraperformanceliquidchromatography AT kyoungwoncho allelicanalysisoftheiglib1ilocusinhexaploidwheatusingreversephaseultraperformanceliquidchromatography |