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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

Inactive Publication Date: 2017-01-04
DALIAN OCEAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this inactivation method is simple and feasible, when the nucleus of the egg is destroyed by radiation, the cytoplasm and other organelles will also be damaged to varying degrees, resulting in the failure of normal development after fertilization.

Method used

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  • Method for obtaining takifugu rubripes androgenesis haploids
  • Method for obtaining takifugu rubripes androgenesis haploids
  • Method for obtaining takifugu rubripes androgenesis haploids

Examples

Experimental program
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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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Abstract

The invention discloses a method for obtaining takifugu rubripes androgenesis haploids. The steps of the method are as follows: (a) the spawning of male takifugu rubripes and female takifugu rubripes with well-developed gonads is induced, the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes are respectively taken, and the sperms and the ova are mixed to form fertilized ova; (b) the androgenesis of the takifugu rubripes is induced by cold shock. The method adopts the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes to form the fertilized ova; cold shock is used for processing (30 to 60 minutes of immersion in ice water of 2 DEG C to 4 DEG C 5 to 8 minutes after fertilization), the egg nuclei and second polar bodies of the takifugu rubripes are induced to release together, and thereby fertilized ova which only have sperm nuclei are formed. Because the genetic inactivation of ova is not required, a complex operation procedure is avoided, the method is cheap and easy to operate, the problem that conventional physiochemical induction can injure fertilized ova is solved, the androgenesis rate is increased, and the method is suitable for use in mass production.

Description

technical field [0001] The invention relates to a new method for obtaining androgenic haploids of the red-finned puffer, in particular to a method for rapidly and efficiently producing the androgenic haploids of the red-finned puffer without egg inactivation. Background technique [0002] Redfin Pufferfish ( Takifugu rubripes ) is a rare seawater fish with the largest individuality, the least toxicity and the highest economic value in the order Pufferiformes. Its meat is white, tender, and delicious, and enjoys the reputation of "the king of fish". It has always been favored by Japan, South Korea and the Chinese, and it is the first choice for marine aquaculture in northern my country and an advantageous aquatic product for export. Although there is not much difference in growth rate between male and female pufferfish, sexually mature male fish have good meat quality and delicious testicles, so the price of mature male fish is twice that of female fish. Therefore, if it i...

Claims

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Application Information

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IPC IPC(8): A01K61/00
CPCY02A40/81A01K61/00
Inventor 周贺姜志强李雅娟江振华徐其征张蕊庄子昕梁雨婷
Owner DALIAN OCEAN UNIV
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