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!
|
| Summary: | 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. |
|---|---|
| ISSN: | 1420-3049 |