Creation and confirmation of heterozygous clones in Japanese flop, Paralichthys olivaceus by microsatellite marker

Abstract

Mitotic gynogenetic diploids Japanese flounder Paralichthys olivaceus were produced by activating eggs with ultra
violet (UV) irradiated sperm of red sea bream (Pagrus major), followed by hydrostatic pressure treatment to block
the first mitotic division. By crossing two mitotic gynogenetic diploid females with two males, two heterozygous
clones of Japanese flounder were produced. Microsatellite marker was used to confirm the genetic status of
maternal parents and their progenies. 20 polymorphic microsatellite markers were chosen, covering 16 out of the
total 24 linkage groups. The four maternal parents used in this experiment were completely heterozygous, while four
mitotic gynogenetic diploids were homozygous for each marker. The genotypes of heterozygous clone progenies
were identical and the combination of parental alleles, demonstrates the successful development of cloning.