Sulforaphene, an isothiocyanate, converted from glucoraphenin in radish, and plays an important role as plant defensive compounds and cancer-preventing agents for humans. In this study, the optimized HPLC method was used to detect the sulforaphene content of fleshy roots among 93 accessions of radish germplasm, and the distribution of sulforaphene content in different origins and different types of radish germplasm was analyzed. The results indicated that the content of sulforaphene among the gemplasm was significantly different. The content of sulforaphene ranged from 34.445 mg/kg﹒DW to 1446.9 mg/kg﹒DW. The highest content is almost 42 times more than the lowest one. The average content of the red skin white flesh and green skin green flesh radish germplasm was high. The average sulforaphene content of the radish germplasm in the eastern China was higher than those in other regions. Two radish germplasm of high sulforaphene content were screened out, which are good materials for further studies.