Taking Eustoma grandiflorum cultivars ‘Double Mariachi Pink’ and ‘Excalibur Blue Picotee’ as experimental material,we extracted genomic DNA from the leaves.The optimized AFLP reaction system was established,after EcoRⅠ/MseⅠ double enzyme digestion,ligation,pre-amplification and selective amplification.154 polymorphic bands were acquired with 64 primer combinations commonly used,from which 4 selective amplification primer combinations(E-ACA/M-CTC,E-ACC/M-CAC,E-AGC/M-CTT,E-ACT/M-CAG) with more polymorphic amplified bands were selected.The average percentage of polymorphic bands was 24.36%.Using the 4 selective amplification primer combinations,through the optimized reaction system,the AFLP fingerprints of 7 common E.grandiflorum cultivars were constructed.Subsequently,the AFLP fingerprints were transformed into the specific digital fingerprints of 28 digits,based on the counting of the amplified bands in the 7 sections of 1 000~300 bp interval of the 4 primer combinations,greatly facilitating the comparison and identification of the germplasm.The genetic similarity coefficient between the cultivars ranged 0.683 5~0.860 8,and the average value was 0.774 6.The results of that study have laid an important foundation for the further research on E.grandiflorum.