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 being vigorously 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 acetic acid based A-PAGE method with urea utilizes commercially available reagents and materials to make gel casting simpler and more efficient.•It can be used to phenotype different wheat varieties to establish purity and to identify mutants of wheat with altered gliadin protein profiles.
Highlights
Method detailsThis method modified a previously published protocol [1] on the preparation of acidic-polyacrylamide gel electrophoresis (A-PAGE) gels used to purify and characterize anti-bacterial peptides and adapted it to separate gliadins in wheat
high-molecular glutenin subunit (HMW-GS) and low-molecular weight glutenin subunit (LMW-GS) genes are encoded by the Glu1 and Glu3 loci that map to the chromosome 1 group long and short arm, respectively [5]
The omega- and gamma-gliadins are encoded by the Gli1 loci that are tightly linked to LMW-GS Glu3 loci in the short arms of the chromosome 1 group
Summary
Modified acid-PAGE method for rapid screening and phenotyping of wheat gliadin mutant lines. A USDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, U.S.A. Acid-polyacrylamide gel electrophoresis (A-PAGE) is used to phenotype different varieties of wheat based on their gliadin profiles. No commercial A-PAGE gels are currently available for cereal researchers to use for separating wheat gliadins. Method name: Acid-PAGE Keywords: A-PAGE, Wheat, Gliadin, Mutant, Acid-PAGE, Gluten Article history: Received 25 September 2019; Accepted 7 March 2020; Available online 20 March 2020.
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