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Small RNAs (sRNAs) control a wide range of development and physiological pathways in plants. To address the response of sRNA biogenesis to drought stress, we identified sRNA biogenesis genes, including 11 encoding argonautes (AGO), 8 encoding Dicer-like proteins (DCL), and 9 encoding RNA-dependent RNA polymerases (RDR) in the peach genome. Notably, the largest numbers of sRNA biogenesis genes are located to chromosome 1. The PAZ, PIWI, and MID domains were identified in PpAGOs, while the ribonuclease IIIa and IIIb domains were characterized in PpDCLs. The RDRP domain was recognized in PpRDRs. Orthologous similarity and collinearity analyses between Arabidopsis and peach revealed 5, 1, and 2 collinear blocks in AGOs, DCLs, and RDRs, respectively. Moreover, 41, 40, and 42 cis-acting elements were located in the promoters of PpAGOs, PpDCLs, and PpRDRs, respectively, with the majority related to drought stress response. Analysis of RNA sequencing (RNA-seq) data revealed that sRNA biogenesis genes were involved in drought stress response in different tissues. Furthermore, the expression of candidate genes was verified in two peach cultivars, Beijing 2-7 (BJ2-7) and Sinai (SN), which are tested as drought-tolerant and sensitive cultivars, respectively, based on the physiological and biochemical analyses, which revealed that the Chinese peach cultivar 'BJ2-7' exhibits greater drought resistance compared to the Egyptian peach cultivar 'SN'. Interestingly, the expression of PpAGO2b, PpDCL2b, PpDCL4, and PpRDR4 genes was induced in 'BJ2-7' but inhibited in 'SN' under drought stress. Overall, this study provides insight into the roles of sRNA biogenesis genes in response to drought stress in peach.