Microsatellite DNA markers were used to investigate the genetic diversity and population structure of Pacific abalone Haliotis discus hannai collected from six locations (Uljin, Ulsan, Daechon, Taean, Wando, and Yosu) where hatchery-produced abalone have been released intensively. There was no distinguishable difference in the observed and expected heterozygosities between the six populations and a cultured population. However, there was a difference in the number of alleles per locus: 12.8 for the cultured population and 13.8 to 15.8 for the six populations. The proportion of stocked abalone ranged from 41.1 to 92.7% for wild-caught populations with a decreasing tendency of alleles per locus for an increasing proportion of stocked abalone. A departure from Hardy-Weinberg equilibrium (HWE) assessed using the Markov chain procedure (P<0.05) was observed in the six populations and cultured population at loci Hdh145 and Hdh5l2. The pairwise Fst test (P<0.05) showed a significant difference between the Uljin and Ulsan populations and four remaining populations (Wando, Daechon, Yosu, and the cultured population), among which the Wando population differed less than the other three populations (Daechon, Yosu, and the cultured population).