Fusarium head blight (FHB) caused by the fungal pathogen Fusarium graminearum is one of the most devastating diseases in barley. However, effective resistance has not been identified in barley germplasm. To enhance barley resistance to FHB, we used host-induced gene silencing (HIGS) to target the F. graminearum histone acetyltransferase gene FgGCN5 in the present study. In the loss-of-function Fggcn5 mutant (ΔFgGCN5), acetylation levels of histone H3 were significantly decreased at several specific lysins, leading to a genome-wide differential expression and impaired metabolic processes affecting pathogenicity of F. graminearum. Using Agrobacterium-mediated gene transformation, we have generated transgenic plants and selected homozygous transformants in the late generations.   Despite demonstrated production of small-interfering RNAs (siRNAs) homologous to FgGCN5 in the transgenic barley; the disease severity, DON accumulation, and fungal biomass showed no significant difference from wild-type. In line with these observations, quantitative revere transcription PCR (qRT-PCR) analysis showed the expression levels of FgSCN5 were not affected by the HIGS construct in the transgenic plants, indicating an inefficiency of the generated siRNAs on silencing the target gene. This research allows for more in-depth analysis for the use of HIGS against FHB. Follow-up investigations with more independent transgenic lines are ongoing to address the incompetence of HIGS targeting FgSCN5 to provide FHB resistance.