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Video

Biotechnology

Session:: Gene Editing Technologies

CRISPR-Cas mediated genetic manipulation of lipoxygenase, precursor fatty acids and flavour components in yellow pea

Tuesday, May 2, 2023
9:10 AM – 9:30 AM MDT
Room: 402
Abstract: Pea (Pisum sativum L.) is one of the major pulse crops grown in Western Canada. As has been clearly shown by Canadian farmers, pulses provide important economic opportunities to boost income and reduce risk through crop diversification. Canada is the leading producer and exporter of field pea in the world. However, the off flavor and anti-nutritional quality are considered as barriers to increased use of pulse products in the food industry. Pea flavor issues continue to plague the food industry and pose a barrier to increased use of pea and other pulses in new food products. Lipoxygenases (LOX), a class of iron-containing enzymes that oxidize polyunsaturated fatty acids, negatively influence the taste and smell of pea products by producing various aldehydes and alcohols. Enhancement of protein flavour and quality is therefore, a priority for new and expanded markets for pea and other pulses. As the LOX is one of the major contributor to volatile organic compounds (VOCs)-associated off-flavours in pulse products, CRISPR/Cas9-mediated gene editing technology was used to create targeted knockout of the pea LOX gene. We obtained eight homozygous mutants using two sgRNAs. We found that the specific LOX activity in three of the edited lines decreased from 250 U/mg protein to 2-23 U/mg. Our results showed that the two precursors of LOX - linoleic and linolenic acid content were unused and significantly higher in the mutant compared to that of the wild type. Two of the abundant volatiles- hexanal and dimethyl sulfide also decreased in mutant lines compared to the wild type pea seeds. In the greenhouse the mutant lines didn’t have any adverse phenotype compared with wild type. It provides at least three new materials for the selection and breeding of low LOX pea lines in the elite background.