Fusarium head blight (FHB) is a major fungal disease affecting small grain cereals worldwide, caused by members of the Fusarium genus. FHB results in significant yield losses annually and poses a serious food safety risk due to contamination of grains with mycotoxins such as deoxynivalenol (DON), a type B trichothecene. DON is a potent inhibitor of protein biosynthesis and induces ribotoxic stress, a response that remains poorly understood in plants. While barley shows strong transcriptional responses to Fusarium infection, it is unclear how these are reflected at the proteomic and metabolomic levels. We aim to understand the molecular defence mechanisms of barley against Fusarium, with a focus on the ribotoxic stress response in planta. For this we use a multiomics approach, integrating transcriptomic, proteomic, and metabolomic analyses. Through analysing the molecular responses of the host plant to infection with both DON-producing and non-producing Fusarium strains, as well as direct exposure to DON, we hope to dissect the specific effects of the mycotoxin versus the pathogen. This integrative strategy will provide novel insights into the molecular dynamics of barley’s defence responses and identify Fusarium- or DON-sensitive metabolic pathways.