Abstract: Bermudagrass are one of the most important and abundant warm-season turfgrass in the world. They are same on morphology between different species or varieties, and could not be identified accurately by traditional morphological method or field district test method. Inter-simple sequence repeat (ISSR) is considered to be the most accurate and efficient molecular marker technology currently on variety identification. In this study, we used ISSR on 27 Bermudagrass individuals to identify genetic diversity and construct Bermudagrass fingerprints of 9 accessions. Thirteen primers produced 54 polymorphic bands, of which 34 bands were found to be polymorphic, 4.1 bands per primer on average. The percentage of polymorphic locus was 64.15% on average. The Nei’s genetic similarity coefficient changed from 0.547 to 0.962, the range was up to 0.415. These demonstrated that test material had ample genetic diversity. Cluster analysis showed that all the accessions could be divided into five groups at GS 0.806: Blackjat and Nan jing Bermudagrass clustered into the first group, Riviera and Common Bermudagrass clustered into the second group, Yang jiang Bermudagrass clustered into the third group, Regent clustered into the forth group and Tifway (T-419), Tifeagle, Tifdwarf clusterd into the fifth group. Research showed that 9 Bermudagrass varieties could be identified by fingerprints based on primer UBC810 and UBC857. The study offered theoretical and technical foundation for research genetic diversity and variety identification using ISSR technology, and could be widely used on variety identification, germplasm resources protection, hybrid parents selection, development and utilization.