High-throughput quantification of amylose and characterization of high-amylose oat lines

• Optimization of a high-throughput amylose quantification assay for oats. • Screening of 481 Ethyl Methane Sulfonate (EMS) mutagenized oat lines for amylose content. • Identification of four oat lines with elevated amylose levels. • Biochemical characterization of one selected high amylose oat line (LT1138) • Novel high-amylose oats varieties with enhanced nutritional benefits ideal for functional foods can now be developed. Oats ( Avena sativa L. ) are gaining increased attention for their rich dietary fibres, proteins and bioactive compounds. Among fibres, starch components denoted as resistant starch (RS) contribute significantly to the functional and nutritional properties of oat-based foods. This study presents the optimization of a high-throughput quantification method for amylose, a key structural component that influences the formation of RS, and the application of this method in the screening of an ethyl methanesulfonate (EMS) mutagenized oat population. By quantifying starch and amylose content in flour obtained from 481 mutant lines, four lines (LT0275, LT0340, LT1132, and LT1138) with amylose contents up to ca 8% higher than the reference variety Belinda were identified. Further characterization of one selected high-amylose line (LT1138) revealed an altered RS structure. Our findings highlight the potential of high-amylose oats in the development of functional foods with enhanced nutritional and health promoting properties.