CRISPR/Cas9-mediated lipoxygenase gene-editing in yellow pea leads to major changes in fatty acid and flavor profiles

Introduction Although pulses are nutritious foods containing high amounts of protein, fiber and phytochemicals, their consumption use in the food industry have been limited due to formation unappealing flavors/aromas described as beany, green, grassy. Lipoxygenase (LOX) enzymes prevalent among pulse seeds, activity can lead specific volatile organic compounds (VOCs) from certain polyunsaturated fatty acids (PUFAs). As a widespread issue legumes, including soybean, these VOCs linked taste perception processed seeds. Methods To address this problem pea proof principle promote wider pulses, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) construct was designed create null alleles (knockouts) PsLOX2 which had implicated generation peas. Results discussion Successful CRISPR/Cas9-mediated LOX gene editing stable transgenic lines (TGP) confirmed by DNA sequencing wild type (WT) TGP pslox2 mutant lines. These were also assessed for activity, PUFA levels, VOCs. Compared WT peas, showed significant reduction (p < 0.05) concentration key VOCs, hexanal, 2-hexenal, heptanal, (E)-2-heptenal, (E,E)-2,4-heptadienal, 1-octen-3-ol, octanal, (E)-2-octenal (E,E)-2,4-nonadienal furan-2-pentyl. The content two essential PUFAs, linoleic α-linolenic acids, known substrates plants, higher flours, indicating efficacy CRISPR-mediated minimizing oxidation further modification PUFAs products. collection headspace ground using portable eNose distinguished Multiple regression analysis that correlated with heptanal (E,E)-2,4-heptadienal flours. Partial Least Squares Regression (PLS-R) plot selected sensor responses distinct clusters Together data demonstrates utility CRISPR mediated mutagenesis quickly improve aroma acid (FA) profiles seeds an elite Canadian variety.