Revolutionizing Yellow Pea: CRISPR/Cas9 Editing Reshapes Fatty Acid and Flavor Dynamics

Although pulses are packed with protein, fiber, and phytochemicals, their utilization in the food industry is constrained by the development of unattractive flavors such as beany, green, and grassy notes, attributed to the prevalence of lipoxygenase (LOX) enzymes in pulse seeds. These enzymes catalyze the production of distinct volatile organic compounds (VOCs) from specific polyunsaturated fatty acids (PUFAs), a common issue in legumes like soybean, contributing to unfavorable taste perceptions in processed pulse-based foods. To address this problem this study focuses on using CRISPR/Cas9 gene editing to target PsLOX2 in peas, aiming to reduce VOCs and improve their flavor and fatty acid profiles. The successful gene editing of stable transgenic pea lines (TGP) was confirmed, leading to decreased LOX activity and concentrations of key VOCs compared to wild type (WT) peas. The edited lines also showed higher levels of essential polyunsaturated fatty acids (PUFAs), linoleic, and α-linolenic acids. Analysis using a portable eNose distinguished between TGP and WT lines based on VOCs. Regression analysis linked LOX activity with specific VOCs in pea flours. Partial Least Squares Regression (PLS-R) analysis showed distinct clusters for WT and TGP lines based on selected PUFAs, VOCs, and sensor responses. Overall, CRISPR-mediated mutagenesis of PsLOX2 was effective in enhancing aroma and fatty acid profiles in elite Canadian pea varieties, highlighting the potential of gene editing to improve pulse utilization in the food industry. The upcoming steps will involve conducting field trials and ingredient testing to evaluate the protein quality and flavor profile of the low lipoxygenase pea lines.