摘要
Tetraploid fish are important for mass production of triploids in polyploid breeding.In this study,we reported a novel protocol for artificial induction of tetraploidy by a combination of cold shock and hydrostatic pressure administered to gynogenetically developed eggs in Japanese flounder Paralichthys olivaceus.The induction was carried out by activating the eggs with UV-irradiated sperm of red sea bream Pagrus major,administering a cold shock(0℃,5 to 45 min)3 min after fertilization to inhibit second polar body exclusion,incubating the eggs for 60 min at 17℃,and treating them with a 650 kg/cm2 hydrostatics pressure shock for 6 min.We named the embryos gyno-tetraploids that developed from eggs after such treatments.The hatching rate of the gyno-tetraploids ranged from 20.99%±3.66%to 36.01%±2.79%,and the tetraploid rate ranged from 80.00%to 100.00%.All-maternal inheritance was verified using 6 high-recombination-rate microsatellite markers.This method successfully induced gyno-tetraploidy.The successful induction of gyno-tetraploidy lays the foundation for triploidization of new varieties with improved economic traits of interest that can benefit commercial culture.
Tetraploid ?sh are important for mass production of triploids in polyploid breeding. In this study, we reported a novel protocol for arti?cial induction of tetraploidy by a combination of cold shock and hydrostatic pressure administered to gynogenetically developed eggs in Japanese ?ounder Paralichthys olivaceus. The induction was carried out by activating the eggs with UV-irradiated sperm of red sea bream Pagrus major, administering a cold shock(0℃, 5 to 45 min) 3 min after fertilization to inhibit second polar body exclusion, incubating the eggs for 60 min at 17℃, and treating them with a 650 kg/cm 2 hydrostatics pressure shock for 6 min. We named the embryos gyno-tetraploids that developed from eggs after such treatments. The hatching rate of the gyno-tetraploids ranged from 20.99%±3.66% to 36.01%±2.79%, and the tetraploid rate ranged from 80.00% to 100.00%. All-maternal inheritance was veri?ed using 6 high-recombination-rate microsatellite markers. This method successfully induced gyno-tetraploidy. The successful induction of gyno-tetraploidy lays the foundation for triploidization of new varieties with improved economic traits of interest that can bene?t commercial culture.
基金
Supported by the Central Public-Interest Scientific Institution Basal Research Fund,CAFS(No.2016HY-ZD0202)
the China Agriculture Research System(No.CARS-47)
the Applied National Science and Technology Support Program in China(No.2012BAD26B01)
