In this research, the first interspecific genetic linkage map of Viola tricolor and V. cornuta was constructed with 66 individuals of F₂ population crossed between V. tricolor and V. cornuta using SRAP marker technique for genotyping. The results revealed that: (1) fifty SRAP primer pairs with more polymorphic, clear and stable bands were selected from 256 primer combinations. (2) A total of 118 SRAP polymorphism markers were obtained after PCR amplification of the F₂ segregation population with 24.6% distorted marker, which met the requirements for genetic map construction. (3) The interspecific genetic map of V. tricolor and V. cornuta was successfully constructed, consisting of 15 linkage group with 67 SRAP markers, and covering the genome of 327.9 cM. The length of linkage groups ranged from 1.6 cM to 52.2 cM, and the average distance between markers is 4.9 cM. This study laid the foundation for high resolution map construction, gene mapping and molecular markerassisted selection in V. tricolor and V. cornuta breeding.