The artificial induction of gynogenesis is a form of chromosome manipulation that has several applications, including the rapid establishment of inbred lines or strains. The technique offers a high degree of homozygosity, sex-control, and accelerated elimination of recessive deleterious genes from a cultured population. We produced two meiotic gynogenetic Japanese flounder () family lines: meio-G1 (the first generation) and meio-G2 (the second generation). We evaluated the efficacy of artificial meiotic gynogenesis for producing pure inbred lines by quantifying the homozygosity and genetic similarity of meio-G1 and using microsatellite markers The average homozygosity among 15 loci was 0.124 7 for meio-G1 and 0.221 5 for meio-G2, which was higher than that in the natural mating family (0.091 7). The average similarity indices among individuals within meio-G1 and meio-G2 were 0.891 7 and 0.923 8, which were higher than within the natural mating family (0.5603). The average similarity in meio-G2 was slightly higher than in meio-G1. The observed heterozygosity in , were homozygous in meio-G1 and meio-G2. Our results suggest that artificially induced meiotic gynogenesis is an efficient method for inbreeding to purify the genome, increase genetic similarity, and c traits in the Japanese flounder. The meiogynogenetic families cultured in our study may be used in further selective breeding research.