Method for creating jade rabbit pleated skirt Taishi goldfish

A technology of Thai lion goldfish and pleated skirt, applied in the field of fish molecular design and breeding, can solve the problems of weakening, loss and unrealization of the target fish traits

Active Publication Date: 2021-05-14
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, hybridization will weaken or even lose many of the original traits of the target fish, and cannot achieve the transfer of single-purpose traits

Method used

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  • Method for creating jade rabbit pleated skirt Taishi goldfish
  • Method for creating jade rabbit pleated skirt Taishi goldfish
  • Method for creating jade rabbit pleated skirt Taishi goldfish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Efficient breeding of pleated skirt Thai lion goldfish

[0036] S1. Seed cultivation: Raise freshly opened fry in a turnover box with a water temperature of 23-25°C and feed live Artemia 3 times a day. The turnover box is 60-100cm long, 40-60cm wide, 20-30cm high, and the water level is 15 -20cm is appropriate. Install a sponge filter on one side of the turnover box to filter, and place an oxygen increasing head next to it for oxygenation, and the air volume is appropriate. Clean the sponge filter once a week.

[0037] After the fry exceed 2cm in length, look down to select four-tailed fry, and then look sideways to remove the deformed and folded fry, and put the selected high-quality fry into a new container for rearing, with a density of about 2 fish per liter of water; the container is equipped with The 7-watt small water pump adds water by trickle, and the daily water intake is about 1 / 5 of the total water body. The three-layer turnover box is filtered ...

Embodiment 2

[0047] Example 2: Exploration of Gynogenetic Conditions

[0048] Two methods of gynogenesis were established, the proportion of normal fish produced by the two methods was compared, and the one with a higher rate of homozygosity was selected as the best gynogenesis condition for Thai lion goldfish.

[0049] 1. Meiotic gynogenesis

[0050] Take carp sperm to inactivate, mix inactivated sperm with pleated skirt Thai lion goldfish eggs, and then perform cold shock treatment. The cold shock treatment method is to develop fertilized eggs at a constant temperature of 24°C for 3-7 minutes, and then place them at 0°C Cold shock was carried out in ice water for 2 minutes to inhibit the discharge of the second polar body, so that the two gametes merged into one to form a tetraploid. After the cold shock treatment, the fertilized eggs hatched into meiotic Thai lion goldfish with jade rabbit traits.

[0051] 2. Mitotic Gynogenesis

[0052] Take carp sperm to inactivate, mix the inactiv...

Embodiment 3

[0057] Example 3: Creation of the Jade Rabbit Pleated Skirt Thai Lion Goldfish

[0058] S1. Target design: Sequence the gene of the pleated skirt Thai lion goldfish to obtain the DNA sequence and amino acid sequence of the gene that regulates the traits of the jade rabbit, and use the wildcard GG[G / A]? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? GG or CC? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? [T / C] CC designs a suitable knockout target on the exon of the DNA sequence of the gene that regulates the traits of the jade rabbit, builds a local genome database after obtaining the target, and uses blast software to search and verify the uniqueness of the target , after determining the uniqueness, the target sequence will be transcribed into gRNA in vitro; "?" in the wildcard character represents an arbitrary base, "[]" represents a match with any base in [], and " / " represents "or".

[0059] The gene regulating the traits of the jade rabbit is the oca2 gene, and th...

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Abstract

The invention relates to a fish molecular design breeding technology, in particular to a method for creating jade rabbit pleated skirt Taishi goldfish. The method comprises the steps of target spot design, microinjection and fertilization incubation. On the basis of a CRISPR / Cas9 gene editing technology, after a pigment gene oca2 is knocked out, jade rabbit goldfish with red eye characteristics is generated on F0-generation goldfishes, efficient gynogenesis conditions and high-temperature group expansion conditions of pleated skirt Taishi goldfish are determined through experiments, and jade rabbit pleated skirt Taishi goldfish homozygotes are obtained on F1-generation goldfish; and through temperature-induced reversion, the F2 generation is massively expanded. By means of molecular breeding, transfer of the single-purpose character is rapidly achieved, the jade rabbit pleated skirt Taishi goldfish which is the variety with the high ornamental value is stably and rapidly created, and wide application prospects and market values are achieved.

Description

technical field [0001] The invention relates to fish molecular design breeding technology, in particular to a creation method of jade rabbit pleated skirt Thai lion goldfish. [0002] technical background [0003] Goldfish originated from the tetraploid crucian carp in my country. It is an ornamental fish formed after hundreds of years of artificial selection, and it can be called the quintessence of our country. In the history of goldfish development, due to mutations, the body color, body shape and almost all external organs of goldfish have undergone obvious changes. Under the action of artificial selection, these mutations have been retained, and these mutations have been crossed and recombined to form multiple compound variations. goldfish; today, there are more than 300 species of goldfish. [0004] Color is an important direction for goldfish breeding in the future. Good-looking colors will greatly increase the ornamental value of fish, such as tiger stripes, ink and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K61/10A01K67/027C12N15/89C12N15/12
CPCA01K61/10A01K67/0276C12N15/89C07K14/461A01K2207/12A01K2217/077A01K2217/206A01K2227/40Y02A40/81
Inventor 余鹏桂建芳汪洋周莉李志李亮亮杨小丽
Owner INST OF AQUATIC LIFE ACAD SINICA
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