Abstract: Different species of Rhodeinae have been wildly distributed across China. Different species have shown very similar morphological characteristics, making their distinguish to be indeed difficult. In order to compare the morphological differences among Rhodeus sinensis, Acheilongnathus tonkinensis, A. macropterus, A. chankaensis and A. taenianalis by the morphometric characteristics, and enrich the classification features, multivariate morphometrics were used to analyze the morphological variations of five Rhodeinae species in Huaihe River Basin (Henan province, China). The results showed that there were significant differences in the body’s width/body’s length, head’s length/body’s length, snout’s length/body’s length, caudal peduncle’s length/body’s length, dorsal fin base’s length/body’s length, pectoral fin’s length/body’s length, pelvic fin’s length/body’s length, and head’s length/snout’s length among the five species (P<0.01), and there were significant differences in the total length/body’s length and body’s height/body’s length (P<0.05). Therefore, these differences mainly focus on the head, tail, fin, and so on. Principal component analysis, discriminant analysis, and cluster analysis showed that the five species had obvious morphological differences, the most significant difference betweenA. taenianalis and A. tonkinensis was found, and the most insignificant difference was observed between A. taenianalis and A. macropterus. The discriminant accuracy rate of the five species varied from 82.35% to 100%, effectively realizing the distinguish of the five Rhodeinae species.