C-Terminally Encoded (CEP) peptides are crucial plant growth regulators. Nevertheless, their physiological roles in cucumber (Cucumis sativus L.), an essential worldwide economical vegetable, remains untapped. In this study, 6 cucumber CEP (CsCEP) genes were identified. A comprehensive analysis showed that the CsCEP proteins displayed conserved characteristics to the identified CEP protein members in other species. CsCEP genes expression levels were variant in cucumber tissues, and were also differentially induced by several environmental factors, suggesting distinct and overlapping roles of CsCEPs in various cucumber developmental processes. We further revealed that synthetic CsCEP4 peptide promoted cucumber primary root growth in a reactive oxygen species (ROS) dependent manner. Overall, our work will provide fundamental insights into the crucial physiological roles of small bioactive peptides during cucumber root development.