To elucidate the role of N-glycosylation in the response of horticultural crops to viral infections, this study employed a comprehensive analytical approach. N-glycopeptides were selectively enriched using zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC), followed by detailed characterization of N-glycosylation patterns in N.benthamiana before and after Tomato chlorosis virus (ToCV) infection.