Objective To analyze and evaluate the genetic quality of closed populations and inbred population of tiger barb,Puntius tetrazona. Method The microsatellite sequences with abundant polymorphism in tiger barb were screened,and two multiple PCR reaction systems were established. Then four populations were analyzed by capillary electrophoresis to identify the genotype and evaluate the genetic diversity. Result The average number of alleles in the wild group,WT closed group,BT closed group,and inbred group were 6. 5833,3. 1667,3. 0833,and 3. 1818. The average polymorphic information contents were 0. 5969,0. 3748,0. 4159 and 0. 4241,respectively. The analysis of population genetic quality showed that the result of genetic differentiation between the four groups were consistent with the process of the inbred population bred from the wild and closed group gradually. Conclusion These microsatellite markers can be used to analyze the genetic quality of different populations of tiger barb,providing a method ological basis for their genetic quality controland monitoring.