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Objective: To study the effect of RNA interference (RNAi) on WD101 gene and its effect on the expression of WD101 mRNA and protein in Schistosoma japonicum. Methods: Double-stranded RNA (dsRNA) WD101 gene and control gene (lacZ) were generated by in vitro transcription and transfected into mechanically transformed schistosomula. The total RNA and protein were isolated simultaneously using TRIzol reagent. The expression levels of mRNA and the protein were determined by quantitative real-time PCR (qPCR) and West blotting, respectively. After injected dsRNA-electroporated schistosomula into BALB/c mouse six weeks, the male and female reproductive organs were observed and measured under the confocal laser scanning microscope. Results: After 1, 3 and 5 d of RNAi, WD101 mRNA level was decreased by 15%, 39%, and 58% in experiment group compared to that in control group;meanwhile, WD101 protein level was decreased by 11%, 28%, and 43% in experiment group compared to that in control group. There were significantly more sperms in testicular lobes in experiment group than that in control group, while there were no significant differences in terms of ovary and vitelline glands between two groups. Conclusions: The dsWD101-RNAi can effectively induce suppression of WD101 gene expression at both mRNA and protein levels. WD101 gene might be a reproduction-related gene in Schistosoma japonicum.