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Aldehyde dehydrogenase (ALDH), a common oxidoreductase in organisms, is an aldehyde scavenger involved in various metabolic processes. However, its function in different pathogenic fungi remains unknown. <i>Fusarium graminearum</i> causes Fusarium head blight in cereals, which reduces grain yield and quality and is an important global food security problem. To elucidate the pathogenic mechanism of <i>F. graminearum</i>, seven genes encoding ALDH were knocked out and then studied for their function. Single deletions of seven ALDH genes caused a decrease in spore production and weakened the pathogenicity. Furthermore, these deletions altered susceptibility to various abiotic stresses. <i>FGSG_04194</i> is associated with a number of functions, including mycelial growth and development, stress sensitivity, pathogenicity, toxin production, and energy metabolism. <i>FGSG_00139</i> and <i>FGSG_11482</i> are involved in sporulation, pathogenicity, and SDH activity, while the other five genes are multifunctional. Notably, we found that <i>FGSG_04194</i> has an inhibitory impact on ALDH activity, whereas <i>FGSG_00979</i> has a positive impact. RNA sequencing and subcellular location analysis revealed that <i>FGSG_04194</i> is responsible for biological process regulation, including glucose and lipid metabolism. Our results suggest that ALDH contributes to growth, stress responses, pathogenicity, deoxynivalenol synthesis, and mitochondrial energy metabolism in <i>F. graminearum</i>. Finally, ALDH presents a potential target and theoretical basis for fungicide development.