Cynoglossus semilaevis disease-resistant breeding gene chip and application thereof

A semi-smooth tongue sole and gene chip technology, applied in genomics, DNA/RNA fragments, instruments, etc., can solve the problem of lack of gene chips in the selection and breeding of semi-smooth tongue sole, and achieve the effects of improving the survival rate of offspring, efficient technical means, and speeding up the breeding process

Pending Publication Date: 2020-11-17
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a gene chip for breeding disease-resistant improved varieties of tongue sole, so as to solve the problem of lack of gene chips in the breeding of improved varieties of tongue sole, to make up for the shortcomings of traditional selective breeding methods, to optimize the breeding process and means, and to be semi-smooth tongue sole The aquaculture industry provides breeding techniques for disease-resistant and high-quality varieties, realizes the upgrading of semi-smooth tongue sole breeding technology, and promotes the efficient and rapid development of semi-smooth tongue sole seed industry

Method used

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  • Cynoglossus semilaevis disease-resistant breeding gene chip and application thereof
  • Cynoglossus semilaevis disease-resistant breeding gene chip and application thereof
  • Cynoglossus semilaevis disease-resistant breeding gene chip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: SNP site screening and chip preparation of "Sole Core No. 1" gene chip

[0036] 1. Establishment of reference population and collection of phenotypic data of tongue sole resistant to Vibriosis harvei

[0037] The reference populations of tongue sole resistant to vibriosis harvei are all derived from the family of tongue sole selected for many years by the inventor's research group. The epidemiological investigation found that Vibrio harveyi is the main pathogenic bacteria in tongue sole aquaculture industry, and infected individuals will have symptoms such as rotten skin and rotten tail. In order to improve the ability of semi-smooth tongue sole to resist Vibrio harvei disease, our research group began to carry out the selection and breeding of semi-smooth tongue sole against Vibrio harvei disease through the artificial infection experiment of Vibrio harvei in 2012.

[0038] In 2014, 2016 and 2018, 11,207 tails of 86 families, 3,714 tails of 23 families and ...

Embodiment 2

[0052] Example 2: Evaluation of the Predictive Accuracy of Genomic Selection Methods Using Reference Populations and Microarray Design Loci

[0053] From the high-quality SNPs obtained in Example 1, extract the same SNPs as the chip design sites, use these SNPs to construct the genomic kinship matrix G matrix, and use 5-fold cross-validation to evaluate the prediction accuracy of the GBLUP method. In addition, the predictive accuracy of the BLUP method was also evaluated for comparison with that of the GBLUP method.

[0054] The analysis results showed that the prediction accuracy of GBLUP (0.780) was higher than that of the BLUP method (0.599) (Table 2, Figure 8 ). Therefore, the SNP loci designed by the "Sole Core 1" gene chip can meet the computational needs of genome selection, and can ensure the smooth development of genome selection breeding.

[0055] The specific operation method is as follows (Linux environment):

[0056] 1. Use PLINK software to extract the chip d...

Embodiment 3

[0072] Example 3: Method for using the "Sole Core No. 1" gene chip prepared in Example 1

[0073] Randomly select 24 resequenced fin ray samples of half-smooth tongue sole, and type these samples according to the following steps:

[0074] 1. Preparation of samples to be tested

[0075] About 1 cm of caudal fin rays of half-smooth tongue sole were cut, and the genomic DNA of the test sample was extracted and purified using a DNA extraction kit (Tiangen, Beijing), and then the DNA quality and concentration were detected by 1% agarose gel electrophoresis and ultraviolet spectrophotometer. Observe under the gel imager after electrophoresis, if a single band appears in the DNA, the fragment length is greater than 10kb, the integrity is good, and there is no degradation, the quality of the genomic DNA is considered to be good ( image 3 ). In addition, using a spectrophotometer to detect, if the sample A260 / 280 is between 1.8 and 2.0, A260 / 230 is greater than 1.5, the concentratio...

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Abstract

The invention aims to provide a gene chip for breeding a cynoglossus semilaevis disease-resistant improved variety, solves the problem that a gene chip is lacked in cynoglossus semilaevis disease-resistant improved variety breeding, overcomes the defects of a traditional breeding technology, and provides a novel molecular breeding method for breeding a disease-resistant high-yield high-quality improved variety. The gene chip can be used for SNP typing of cynoglossus semilaevis at the whole genome level, calculating the genetic effect of each SNP site, estimating the genome breeding value (GEBV) of an individual, and screening fishes with high disease resistance according to the GEBV of the individual; and the survival rate of the offspring can be obviously improved by utilizing the optimized parent fishes to breed the offspring. Therefore, by utilizing the gene chip, the breeding process of the cynoglossus semilaevis disease-resistant improved variety can be accelerated, the breeding period is shortened, and an efficient molecular breeding technical means is provided for breeding of the cynoglossus semilaevis disease-resistant improved variety.

Description

technical field [0001] The invention belongs to the technical field of aquatic genetics and breeding, and in particular relates to a gene chip for breeding disease-resistant improved varieties of tongue sole, a preparation method and application thereof. Background technique [0002] Fish farming is a pillar industry in aquaculture. In 2018, the total output of aquaculture in my country was 49.9106 million tons, and the output of fish farming was 26.9378 million tons, accounting for 53.97% of the total aquaculture output. At the same time, fish, as a high-quality protein source, is also an important component of my country's food composition. Semi-smooth tongue sole is a rare and high-quality marine fish cultured in coastal areas of my country. It is loved by consumers for its tender meat and smooth taste. With the gradual expansion of the scale of industrial farming in my country, problems such as frequent disease and germplasm degradation have become increasingly prominen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11G16B20/20
CPCC12Q1/6888G16B20/20C12Q2600/156C12Q2600/124
Inventor 陈松林卢昇周茜陈亚东刘洋王磊李仰真杨英明
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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