Fusarium culmorum causes head blight, produces toxins and reduces yield and quality of cereals. To prevent damage caused by fusarium head blight (FHB), azole fungicides are mainly applied. The occurrence of insensitivity to azoles is a major problem in agriculture. The present study shows that a tebuconazole insensitive strain of F. culmorum can be readily produced in the laboratory, but that the resulting strain of the fungus is of lower fitness in vitro. Insensitivity was confirmed microscopically and by cell viability and metabolic activity. The tebuconazole insensitive strain shows cross insensitivity to nine important azoles. In addition, plants inoculated with the insensitive F. culmorum strain showed no reduction of FHB symptoms and deoxynivalenol (DON) content after tebuconazole treatment, compared to an inoculation with the sensitive strain. Use of wheat cultivars carrying a high resistance level (i.e. cv. Toras) was the most effective method for reducing symptoms and decreasing DON content, independent from the level of fungicide insensitivity of the F. culmorum strain. In conclusion, resistant cultivars and a fungicide mixture which combines different mechanisms of action in fungal metabolism should be applied to avoid fungicide insensitivity of Fusarium spp. in future.