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Chinese tongue sole is a marine fish with ZW sex determination. Genome sequencing suggested that the Z-linked  dmrt1  is a putative male determination gene, but direct genetic evidence is still lacking. Here we show that TALEN of  dmrt1  efficiently induced mutations of this gene. The ZZ  dmrt1  mutant fish developed ovary-like testis, and the spermatogenesis was disrupted. The female-related genes  foxl2  and  cyp19a1a  were significantly increased in the gonad of the ZZ  dmrt1  mutant. Conversely, the male-related genes  Sox9a  and  Amh  were significantly decreased. The  dmrt1  deficient ZZ fish grew much faster than ZZ male control. Notably, we obtained an intersex ZW fish with a testis on one side and an ovary on the other side. This fish was chimeric for a  dmrt1  mutation in the ovary, and wild-type  dmrt1  in the testis. Our data provide the first functional evidence that  dmrt1  is a male determining gene in tongue sole.

Genome editing reveals dmrt1 as an essential male sex-determining gene in Chinese tongue sole (Cynoglossus semilaevis)