GhWRKY33 negatively regulates jasmonate-mediated plant defense to Verticillium dahliae.

Verticillium wilt, caused by Verticillium dahliae, seriously restricts the yield and quality improvement of cotton. Previous studies have revealed the involvement of WRKY members in plant defense against V. dahliae, but the underlying mechanisms involved need to be further elucidated. Here, we demonstrated that Gossypium hirsutum WRKY DNA-binding protein 33 (GhWRKY33) functions as a negative regulator in plant defense against V. dahliae. GhWRKY33 expression is induced rapidly by V. dahliae and methyl jasmonate, and overexpression of GhWRKY33 reduces plant tolerance to V. dahliae in Arabidopsis. Quantitative RT-PCR analysis revealed that expression of several JA-associated genes was significantly repressed in GhWRKY33 overexpressing transgenic plants. Yeast one-hybrid analysis revealed that GhWRKY33 may repress the transcription of both AtERF1 and GhERF2 through its binding to their promoters. Protein-protein interaction analysis suggested that GhWRKY33 interacts with G. hirsutum JASMONATE ZIM-domain protein 3 (GhJAZ3). Similarly, overexpression of GhJAZ3 also decreases plant tolerance to V. dahliae. Furthermore, GhJAZ3 acts synergistically with GhWRKY33 to suppress both AtERF1 and GhERF2 expression. Our results imply that GhWRKY33 may negatively regulate plant tolerance to V. dahliae via the JA-mediated signaling pathway.