Viroids are small, naked, infectious single-stranded RNA molecules that exploit host factors to replicate. Some viroids have been linked to severe diseases in agricultural crops, including the recent outbreak of , previously known as Citrus bark cracking viroid (CBCVd), in hop plants (). Numerous studies have demonstrated the involvement of viroid-derived RNA in viroid pathogenesis through interactions with RNAi host factors, leading to alterations in gene expression, metabolism, and phenotype. Recent research efforts have also focused on elucidating viroid-induced changes in DNA methylation patterns via the RNA-directed DNA methylation pathway. In this study, we conducted an epigenome analysis of CBCVd-infected hop plants to provide novel evidence supporting the putative role of DNA methylation in CBCVd viroid pathogenesis. Our findings revealed that several genes involved in pathogen interaction pathways, such as MAPK signaling and LRR, exhibit hypomethylation, suggesting that their increased transcription enhances the host's ability to counteract the pathogen. Intriguingly, genes associated with RNA transcription and encoding key proteins, such as POL II, POL IV, and POL V, display hypermethylation, highlighting the significance of DNA methylation as a defense mechanism.IMPORTANCEViroids are emerging as a substantial threat to various crops; however, our understanding of the molecular mechanisms governing their pathogenesis and the host's defense remains incomplete. This knowledge gap leaves crop disease management reliant on unsustainable strategies. Our research seeks to address this issue by examining the complex world of infected hop plants. Specifically, we are investigating the DNA methylation processes, providing insights into the less-explored aspects of the host's response to viroid interaction. Our aim was to unravel the complexities of how viroids influence the molecular landscape within plants and the corresponding host defenses. By understanding these interactions, we hope to provide insights that lead to more sustainable ways to protect crops and keep agriculture resilient against viroid-related threats.