Cucumber plants (Cucumis sativus) displaying symptoms of phyllody, sterility, and flower virescence were observed in fields located in the Dianbai district of Guangdong province, China. Total DNA was extracted from the symptomatic plants; this was followed by molecular detection using a set of universal phytoplasma primer pairs, R16mF2/mR1, P1/P7, and SecYF1(II)/SecYR1(II). This resulted in the PCR amplification of products corresponding to expected sizes of 1.4 kb, 1.8 kb and 1.7 kb, respectively. The 16S rDNA sequence obtained exhibited 100% similarity with the eggplant phyllody phytoplasma, the ‘Cleome rutidosperma’ witches’ broom phytoplasma, and the ‘Desmodium ovalifolium’ witches’ broom phytoplasma strain DeOWB, all of which belong to the 16SrII group. Phylogenetic analysis, based on the 16S rDNA gene and SecY gene sequences, confirmed the close affiliation of the detected phytoplasma isolate, tentatively designated as cucumber phyllody phytoplasma (CuPh) China isolate, with the 16SrII-A subgroup. Additionally, virtual restriction fragment length polymorphism (RFLP) analysis of the 16S rDNA sequence revealed a pattern that was identical to that of the 16SrII-A subgroup. This is the first report of cucumber phyllody phytoplasma in China.