Genetic diversity of 50 cucumber (Cucumis sativus L.) accessions from American, Holland, Japan and China was detected and evaluated using RAPD markers. 25 selected primers produced 215 scorable polymorphic RAPD bands and the ratio of polymorphism is 86.98%. Four main groups of cucumber accessions could be distinguished from UPGMA analysis: Occidental cucumber, South-China cucumber, North-China cucumber and Xishuangbanna cucumber. Our result confirmed that cucumber is a narrow-based germplasm, nevertheless RAPD analysis was still useful in cucumber genotypic differentiation. More significantly, Xishuangbanna cucumber (Cucumis sativus L. Var Xishuangbannanesis Qi et Yuan), a mutation of cucumber, was clustered together to a special group in the study, which implied its special status in cucumber germplasm.