Method for obtaining takifugu rubripes androgenesis haploids
A technology for the development of male and female pufferfish, which is applied in fish farming, application, climate change adaptation, etc., can solve problems such as damage, cytoplasmic damage, and abnormal development, and achieves avoiding operating procedures, easy operation, and improving male sex. Effect of nuclear development rate
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Embodiment 1
[0018] The present embodiment provides a method for obtaining the haploid of the redfin puffer androgenesis, and the specific implementation steps are as follows:
[0019] a. Take the well-developed pufferfish (female and male fish) as parents, induce labor with the method of inducing labor in the prior art, lightly press the abdomen of the female fish after 24 hours, and observe whether eggs are discharged. When the eggs are released, lift the fish, squeeze the sides of the abdomen, collect the eggs into a clean basin, and then collect the sperm into the basin. According to the method of prior art, the diluted semen is mixed with ovum to form fertilized ovum.
[0020] b. Place the fertilized eggs in 4°C ice water 8 minutes after fertilization, soak for 30 minutes, and then place them in normal temperature water for incubation. During hatching, the aerated water is frequently replaced according to the hatching method of the prior art, the indoor air is kept fresh and circulat...
Embodiment 2
[0022] This embodiment provides a method for identifying androgenic fry, which includes: 1. Early embryo chromosome analysis to identify androgenic fry; 2. Microsatellite molecular markers to identify androgenic fry. The specific implementation steps are as follows:
[0023] 1. Chromosome analysis and identification of gynogenetic fry:
[0024] The number of chromosomes in somatic cells of puffer puffer in Example 1 and the control group was detected by the early single embryo preparation method. The difference between the control group and Example 1 was that the fertilized eggs were not treated with cold shock.
[0025] The early single embryo production method is as follows: take 30-50 embryos at the stage of developing eye cells in Example 1 and the control group, cut off the egg membrane and yolk with tweezers, and put them in 0.0025% colchicine for 45 minutes, 0.8 % citric acid for 20 min, and then fixed with pre-cooled Carnot's fixative solution (methanol: glacial aceti...
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