Method for inducing gynogenetic diploid of Pseudosciaena crocea
A technology of gynogenesis and large yellow croaker, applied in the field of gynogenesis, to achieve the effect of accelerating homozygosity
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Embodiment 1
[0016] Take the semen of large yellow croaker and dilute it 30 times with the diluent, put it in a petri dish with a diameter of 6-15cm, and irradiate it with ultraviolet rays. The ultraviolet irradiation dose is 50000-100000μW / cm 2 , semen thickness 0.5mm. The preparation of the diluent is: each liter of solution contains 7.5g of NaCl, 0.2g of KCl, and 0.2g of CaCl 2 0.2g, NaHCO 3 0.2g, to inactivate the genetics of sperm. Adjust the seawater temperature to 22-23°C, mix mature eggs of large yellow croaker and sperm of large yellow croaker irradiated by ultraviolet rays in the sea water, and the final concentration of sperm should be at least 10 5 individual / ml to start the gynogenesis of large yellow croaker eggs. 3 to 4 minutes after fertilization, the eggs were transferred to seawater at 0 to 1°C for cold shock for 10 minutes, so that the gynogenetic haploid chromosome group of large yellow croaker was doubled to become gynogenetic diploid. The gynogenetic eggs of lar...
Embodiment 2
[0029] Similar to Example 1, the difference is that the large yellow croaker semen is diluted 10 times with the diluent and then irradiated with ultraviolet rays, and the ultraviolet irradiation dose is 50000-100000 μW / cm 2 , Semen thickness 0.8mm. The preparation of the diluent is: each liter of solution contains 7.5g of NaCl, 0.2g of KCl, and 0.2g of CaCl 2 0.2g, NaHCO 3 0.2g, to inactivate the genetics of sperm. Adjust the seawater temperature to 20-22°C, mix mature eggs of large yellow croaker and sperm of large yellow croaker irradiated by ultraviolet rays in the sea water, and the final concentration of sperm should be at least 10 5 individual / ml to start the gynogenesis of large yellow croaker eggs. 4 to 5 minutes after fertilization, the eggs are transferred to a hydraulic press for hydrostatic treatment at a pressure of 45MPa / cm 2 , The time for hydrostatic treatment in the hydraulic press is 2 to 3 minutes. Doubling the gynogenetic haploid chromosome set of la...
Embodiment 3
[0031] Similar to Example 1, the difference is that the large yellow croaker semen is diluted 50 times with the diluent and then irradiated with ultraviolet rays, and the ultraviolet irradiation dose is 50000-100000 μW / cm 2 , semen thickness 1.0mm. The preparation of the diluent is: each liter of solution contains 7.5g of NaCl, 0.2g of KCl, and 0.2g of CaCl 2 0.2g, NaHCO 3 0.2g, the genetic inactivation of sperm. Adjust the seawater temperature to 23-25°C, mix mature eggs of large yellow croaker and sperm of large yellow croaker irradiated by ultraviolet rays in the sea water, and the final concentration of sperm should be at least 10 5 per ml to start the gynogenesis of large yellow croaker eggs. 4 minutes after fertilization, the eggs are transferred to a hydraulic press for hydrostatic treatment at a pressure of 45MPa / cm 2 , the time of hydrostatic treatment in the hydraulic press is 3min. Doubling the gynogenetic haploid chromosome set of large yellow croaker into g...
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