Method for producing allopolyploid misgurnus anguillicaudatus through inhibiting fertilized egg polar body release
An allopolyploid and tetraploid loach technology, applied in fish farming, application, climate change adaptation, etc., can solve the problem of low induction success rate, achieve obvious weight gain effect, easy access to materials, and strong promotion effect of benefit
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Embodiment 1
[0017] The establishment of embodiment 1 optimal induction condition
[0018] The invention utilizes a physical method (cold shock treatment) to induce the allopolyploid loach, and mainly obtains the allopolyploid loach by inhibiting the release of the second polar body of the fertilized egg. According to the principle of three-factor experimental design, the experimental conditions (shock start time, shock temperature and shock treatment time) were designed in gradient respectively. The shock start time was the 4th, 5th and 6th minutes after fertilization respectively; 1°C, 2°C, 3°C, 4°C; the duration of shock treatment was 25 minutes, 30 minutes, and 35 minutes, respectively. Two parallels were designed for each experimental group, and the appropriate induction conditions were determined for subsequent experiments by counting the survival rate of embryos and the success rate of polyploid induction.
[0019] The preferred start time of shock in the present invention is 5 min...
Embodiment 2
[0020] The establishment of embodiment 2 loach allopolyploid
[0021] First, wild loach was caught from the waters around Wuhan City, Hubei Province. Select well-developed, disease-free and non-injured as breeding parents. In the laboratory, flow cytometry (Beckman, Quanta SC) was used to identify the DNA content of blood cells to determine their ploidy. By injecting oxytocin drugs to the breeding parents, the oxytocin drugs used are ovulation-inducing hormone No. 2 (LRH-A2) and dioconone (DOM). Half, the injection method is a single intraperitoneal injection. After the injection of oxytocin drugs, the male and female fish were kept separately in different water tanks, and the water temperature was 24-26°C. 12-15 hours after the injection of oxytocin drugs, dissect the male fish to take out the semen, and mix the testis and sperm preservation solution (preservation solution formula: sodium chloride 7.5g, potassium chloride 0.3g, calcium chloride 0.3g, glucose 3.0g, distille...
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