Abstract: The aim was to seek appropriate ISSR primers of beet identification and then build beet fingerprint. The author utilized one species to do annealing gradient experiment on 100 ISSR primers and then expanded all primers under their best annealing temperature using 16 imported beet species, appropriate beet species was selected for purity identification. As a result, 16 primers with high level of polymorphism, clear amplification bands and good repetitiveness were selected which could expand to over 100 polymorphic bands in total with PIC over 0.6 on average. Theoretically speaking, these core primers could accomplish the identification of beet species.