Modified acid-PAGE method for rapid screening and phenotyping of wheat gliadin mutant lines

Authors: WATRY, HANNAH - Gladstone Institutes; Zerkle, Alexander; L Chingcuanco, Debbie

Submitted to: MethodsX
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/7/2020
Publication Date: 3/20/2020
Citation: Watry, H., Zerkle, A.J., Chingcuanco, D.L. 2020. Modified acid-PAGE method for rapid screening and phenotyping of wheat gliadin mutant lines . MethodsX. 7. https://doi.org/10.1016/j.mex.2020.100858.
DOI: https://doi.org/10.1016/j.mex.2020.100858

Interpretive Summary: Gluten is the major determinant of the unique properties of wheat dough that enable the production of important food products including breads, cakes, cookies and pasta. A family of proteins known as gliadins are a major component of wheat gluten and are important for the baking quality of the flour, but some of these proteins are also known to trigger human immunogenic responses that lead to several gluten-related health issues like celiac disease and wheat-dependent exercise induced anaphylaxis. A novel screening method was developed to identify wheat plants that produce fewer gliadin proteins. This process enables an easy and efficient way to test for differences in the production of these important gluten proteins in different wheat cultivars and novel wheat breeding lines.

Technical Abstract: Acid-polyacrylamide gel electrophoresis (A-PAGE) is used to phenotype different varieties of wheat based on their gliadin profiles. The family of gliadin proteins is a major component of wheat gluten. Gluten is the major determinant of the unique viscoelastic property of wheat dough that is necessary in the production of important food products including bread, cake, cookies and pasta. However, several gliadin proteins are also known to be causal agents in triggering human immunogenic responses that lead to several gluten-related health risks like celiac disease and wheat-dependent exercise induced anaphylaxis. Therefore, research to identify wheat lines with reduced levels of immunogenic proteins is vigorously being pursued in several laboratories around the world. Unfortunately, no commercial A-PAGE gels are currently available for cereal researchers to use for separating wheat gliadins. This work reports the development of an easy-to-use A-PAGE protocol to resolve gliadins with high reproducibility and resolution to screen and phenotype gliadin deficient lines in wheat. This method can be used to phenotype different wheat varieties to establish purity. This A-PAGE method was used to identify mutants derived from a commercial variety of wheat that are deficient in several members of the gliadin family.