The Asian green mussel, Perna viridis, has been widely cultured in Southeast Asia as delicious seafood depending on natural spat. We analyzed the genetic diversity and population structure of three populations (two from Singapore and one from Malaysia) from natural habitats and two from Singapore farms, by genotyping a total of 262 adult individuals using nine polymorphic microsatellites. In all five populations, high allelic (number of alleles A = 11.0–14.4,allelic richness Ar = 10.4–13.9) and gene diversity (expected heterozygosity He = 0.73–0.77) were observed, whereas deficiency of heterozygosity (inbreeding index f = 0.08–0.12) was detected in all populations. Genetic differentiation among populations was low (pairwise fixation index ranged from 0.003 to 0.088), but statistically significant. The usefulness of the information about genetic variations for genetic resource management of the populations in wild habitats and farms to keep genetic variation and setting up a breeding program of Asian green mussel was discussed.