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Fusarium head blight (FHB) caused by Fusarium graminearum (FG) is a destructive disease impacting barley worldwide. The disease reduces the grain yield and contaminates grains with mycotoxins, such as the trichothecene deoxynivalenol (DON). Although the infection mainly affects the grain yield, little is known about its impact on grain structural and biochemical properties. Yet, such information is instrumental to characterize the facets of resistance in the grains. After artificial inoculation of six barley cultivars with FG in a 2 years field test, different levels of symptoms on spikes, of colonisation of grains and of DON content were observed. The infections caused a reduction in grain weight and an average decrease of 10% of the β- glucan content in grains, indicating alterations of grain filling, composition and structure. According to our results, we postulate the presence of two distinct resistance mechanisms in the grain, tolerance to grain filling despite infection as well as the inhibition of mycotoxin accumulation. Differently to wheat, in barley, type IV resistance (tolerance of the grain to infection) is directly linked with type III resistance (resistance against kernel infection). The resistance against toxin accumulation (named type V resistance in wheat) appeared to be independent to all other resistance types. Generally, the resistance was significantly influenced by the environment and by genotype x environment interactions explaining the generally weak stability of resistance in barley. Interestingly, a significant and inverse relationship between DON contamination and β-glucan content in grains suggests that high β-glucan content in grains contributes to type V resistance.