Summary The vascular wilt fungus Verticillium dahliae is one of the most destructive pathogens of cotton ( Gossypium hirsutum ) and many other economically important dicot plants. Fungal pathogens require Myosin‐mediated actomyosin motility system for colonization of their host plants; however, the mechanisms underlying this process have not been fully characterized for V. dahliae . Here, in a knock‐out experiment, we characte...