To determine the genetic diversity and population structure of sweet potato accessions cultivated in China, and to establish the genetic relationships among their germplasm types, a representative collection of 240 accessions was analyzed using inter-simple sequence repeat(ISSR) markers. The mean genetic similarity coefficient, Nei’s gene diversity, and shared allele distance of tested sweet potato accessions were 0.7302, 0.3167 and 0.2698, respectively. The 240 accessions could be divided into six subgroups and five subpopulations based on neighbor-joining(NJ) clustering and STRUCTURE results, and obvious genetic relationships among the tested sweet potato accessions were identified. The marker-based NJ clustering and population structure showed no distinct assignment pattern corresponding to flesh color or geographical ecotype of the tested sweet potato germplasm. Analysis of molecular variance(AMOVA) revealed small but significant difference between white and orange-fleshed sweet potato accessions. Small but significant difference were also observed among sweet potato accessions from the Southern summer-autumn sweet potato region, the Yellow River Basin spring and summer sweet potato region and the Yangtze River Basin summer sweet potato region. This study demonstrates that genetic diversity in the tested sweet potato germplasm collection in China is lower than that in some reported sweet potato germplasm collections from other regions. Pedigree investigations suggest that more diverse Chinese sweet potato varieties should be formed by broadening the selection scope of breeding parents and incorporating the introduced varieties into future breeding programs. To determine the genetic diversity and population structure of sweet potato accessions cultivated in China, and to establish the genetic relationships among their germplasm types, a representative collection of 240 accessions was analyzed using inter-simple sequence repeat (ISSR) markers. The mean genetic similarity coefficient, Nei’s gene diversity, and shared allele distance of tested sweet potato access were 0.7302, 0.3167 and 0.2698, respectively. The 240 accessions could be divided into six subgroups and five subpopulations based on neighbor-joining (NJ) clustering and STRUCTURE results, and obvious genetic relationships among the tested sweet potato accessions were identified. The marker-based NJ clustering and population structure showed no distinct assignment pattern corresponding to flesh color or geographical ecotype of the tested sweet potato germplasm. Analysis of molecular variance (AMOVA) revealed small but significant difference between white and orange-fleshed sweet po Small but significant difference were also observed among sweet potato accessions from the Southern summer-autumn sweet potato region, the Yellow River Basin spring and summer sweet potato region and the Yangtze River Basin summer sweet potato region. This study demonstrates that genetic diversity in the tested sweet potato germplasm collection in China is lower than that in some reported sweet potato germplasm collections from other regions. Pedigree investigations suggest that more Chinese sweet potato varieties should be formed by broadening the selection scope of breeding parents and incorporating the introduced varieties into future breeding programs.