이정훈 (강원대학교)

권춘탁 (경희대학교)

이지호 (강원대학교)

이정훈 (강원대학교)

서명균 (경희대학교)

송민호 (강원대학교)

최은송 (강원대학교)

권춘탁 (경희대학교)

이지호 (강원대학교)

This study evaluated the metabolic effects of gene knockout in tomato fruit through comprehensive metabolomic profiling. More than 400 metabolites, including primary metabolites, carotenoids, vitamins, and other secondary metabolites, were analyzed in mutant and wild-type fruits. The results indicated that the mutation caused extensive metabolic changes extending beyond the intended target pathway.

PCA and PLS-DA showed clear separation between wild-type and mutant fruits, reflecting distinct metabolic reprogramming. Most metabolic pathways were broadly suppressed in the mutant, whereas sugar-related metabolites accumulated. In particular, depletion of the amino acid pool was associated with reduced activity of the urea cycle, TCA cycle, and GABA biosynthesis. Impaired amino acid metabolism was also linked to decreased glycerophospholipid biosynthesis, suggesting disruption of membrane lipid formation.

Carotenoid analysis showed no significant changes in lycopene derived compounds, including lutein and β-carotene. However, phytoene, phytofluene, and lycopene were significantly reduced, indicating suppression of upstream carotenoid biosynthesis.

Overall, these findings demonstrate that gene knockout can induce broad metabolic perturbations beyond the target pathway and emphasize the need for off-target metabolic evaluation in genetically modified crop development.