The invention provides a breeding method for constructing XX / XY sex determination all-female sterile fish and application. The method comprises the following steps: knocking out a catalytic
enzyme coding
gene of a fish
sex steroid hormone synthesis pathway by using a
gene editing technology to block the fish
sex steroid hormone synthesis pathway, screening to obtain an effectively mutated homozygote XX sex genetic type pseudo male fish, and hybridizing the pseudo male fish with improved allotetraploid crucian and
carp female fish to obtain the fish
sex steroid hormone synthesis pathway. Therefore, an allotriploid
population is obtained, and the
population completely develops into female fishes and is sterile, so that the effect of obtaining all-female and sterile populations is achieved at the same time. The method provided by the invention has very strong applicability, and can be widely applied to
aquaculture of fishes to obtain female unisexual groups of the fishes so as to improve the
aquaculture yield, realize effective control on female
fertility and protect
ecological safety; the method has a wide application prospect in the fields of genetic breeding and
ecological safety of
aquaculture fishes.