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In this study, the disease resistance gene  PlWRKY65  was isolated from the leaves of  Paeonia lactiflora  and analyzed by bioinformatics methods, and the localization of the encoded protein was explored. Quantitative real-time PCR (qRT-PCR) was also used to explore the response of this gene to  Alternaria tenuissima . The results showed that the gene sequence contained multiple cis-acting elements involved in the response to hormone signaling molecules belonging to the IIe subgroup of the WRKY family, and the encoded proteins were located in the nucleus. The  PlWRKY65  gene has a positive regulatory effect on  A. tenuissima  infection. After silencing the  PlWRKY65  gene via virus-induced gene silencing (VIGS), it was found that the gene-silenced plants were more sensitive to  A. tenuissima  infection than the wild plants, exhibiting more severe infection symptoms and different degrees of changes in the expression of the pathogenesis-related (PR) genes. In addition, we showed that the endogenous jasmonic acid (JA) content of  P. lactiflora  was increased in response to  A. tenuissima  infection, whereas the salicylic acid (SA) content decreased. After  PlWRKY65  gene silencing, the levels of the two hormones changed accordingly, indicating that  PlWRKY65 , acting as a disease resistance-related transcriptional activator, exerts a regulatory effect on JA and SA signals. This study lays the foundation for functional research on WRKY genes in  P. lactiflora  and for the discovery of candidate disease resistance genes.

The WRKY transcription factor PlWRKY65 enhances the resistance of Paeonia lactiflora (herbaceous peony) to Alternaria tenuissima