Abstract
The RNA-directed DNA methylation (RdDM) pathway is a crucial epigenetic mechanism governing plant responses to environmental stress. While the RdDM pathway has been extensively studied in Arabidopsis thaliana, the comprehensive understanding of its components in cucumber (Cucumis sativus L.) remains lacking. In this study, we performed a genome-wide identification and characterization of RdDM pathway genes in cucumber, followed by an analysis of their expression patterns across various tissues and under multiple abiotic stress conditions. A total of 67 putative CsRdDM genes were identified, which are unevenly distributed across the cucumber's chromosomes. Phylogenetic and gene structure analyses revealed considerable evolutionary divergence, particularly within the key Argonaute gene family (CsAGO). Crucially, the promoter regions of CsRdDM genes were found to contain cis-regulatory elements associated with abiotic stress, light signaling, and development, suggesting their potential involvement in complex regulatory networks. RT-qPCR assays confirmed that CsRdDM genes exhibit distinct and stress-specific transcriptional patterns. Notably, several genes such as CsAGO4 and CsIDN2 showed antagonistic expression between roots and leaves under drought (PEG-6000) stress, implying a sophisticated, tissue-specific defense mechanism. Among them, CsAGO4 emerged as a candidate gene responsive to abiotic stress. Those findings provide new insights into the regulatory roles of CsRdDM genes under abiotic stress and highlight candidate genes for the genetic improvement of stress tolerance in cucumber.