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Several studies have revealed that actin depolymerizing factors (ADFs) participate in plant defence responses; however, the functional mechanisms appear intricate and need further exploration. In this study, we identified an  ADF6  gene in upland cotton (designated as  GhADF6 ) that is evidently involved in cotton's response to the fungal pathogen  Verticillium dahliae . GhADF6 binds to actin filaments and possesses actin severing and depolymerizing activities in vitro and in vivo. When cotton root (the site of the fungus invasion) was inoculated with the pathogen, the expression of  GhADF6  was markedly down‐regulated in the epidermal cells. By virus‐induced gene silencing analysis, the down‐regulation of  GhADF6  expression rendered the cotton plants tolerant to  V. dahliae  infection. Accordingly, the abundance of actin filaments and bundles in the root cells was significantly higher than that in the control plant, which phenocopied that of the  V. dahliae ‐challenged wild‐type cotton plant. Altogether, our results provide evidence that an increase in filament actin (F‐actin) abundance as well as dynamic actin remodelling are required for plant defence against the invading pathogen, which are likely to be fulfilled by the coordinated expressional regulation of the actin‐binding proteins, including ADF.

GhADF6 ‐mediated actin reorganization is associated with defence against Verticillium dahliae infection in cotton