Method for selecting nervous necrosis virus resistant individuals of epinephelus lanceolatus

A saddle grouper, nerve necrosis technology, applied in biochemical equipment and methods, microbial determination/inspection, climate change adaptation, etc. The problem of long cycle and inability to select disease-resistant parents, etc., achieves significant economic and social benefits, great use value, and high accuracy.

Pending Publication Date: 2022-07-12
SOUTH CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the breeding process, it was found that the saddleband grouper has a long reproductive cycle, and it usually takes 6-8 years to breed one generation. It is not realistic to adopt the traditional breeding method
Therefore, the breeding process can be accelerated by combining the genetic information of the saddle grouper itself. In addition, the disease resistance traits cannot be observed through phenotype, especially the disease-resistant parents cannot be selected for offspring breeding, which makes the selection difficult. Therefore, there is an urgent need for an accurate method that can predict the disease resistance of saddle grouper through genetic information such as SNP markers

Method used

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  • Method for selecting nervous necrosis virus resistant individuals of epinephelus lanceolatus
  • Method for selecting nervous necrosis virus resistant individuals of epinephelus lanceolatus
  • Method for selecting nervous necrosis virus resistant individuals of epinephelus lanceolatus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The method for selecting an individual against nerve necrosis virus of grouper saddle provided by the present embodiment comprises the following steps:

[0033] (1) Obtaining SNP site information of grouper saddle

[0034] Select healthy saddle grouper fry (the size of healthy saddle grouper fry is uniform, the length is 4-6cm, and the number is 600), and RGNNV (red point grouper neuronecrosis virus (RGNNV) is used to isolate from red point grouper. , obtained by conventional laboratory cultivation, and the cultivation method may refer to but not limited to the following literature: Isolation, identification and pathological observation of a carp spring viremia virus from a perch carp source, Zheng Liping, China Fisheries Science, 2019.5; or fish Research on the isolation and partial characteristics of viroid neuronecrosis virus, Jiang Fangjun, Master's thesis of Huazhong Agricultural University, 2008) to challenge it, collect the fin rays or muscles of dead fry every d...

Embodiment 2

[0066] In order to verify the accuracy of the identification method in Example 1, the present embodiment adopts the following tests to verify:

[0067] (1) On June 20, 2020, 110 grouper fry with a size of 3.15±0.23 cm were selected and cultivated by Hainan Chenhai Aquaculture Co., Ltd. At this time, the disease resistance of the fry could not be judged from the appearance. After taking a small amount of caudal fins, use a separate feeding box to raise, and mark the box;

[0068] (2) extracting the DNA of each fish for resequencing to obtain SNP site information of each fish;

[0069] (3) Each fish was challenged while resequencing was performed, and each fish was intraperitoneally injected with 30 μL at a concentration of 2.46×10 7 TCID 50 / mL of virus, record the marking number on the dead fish box;

[0070] (4) According to the resequencing results, the Genome Association and Prediction Integrated Tool (GAPIT3) was used to calculate the breeding value. The number of fish ...

Embodiment 3

[0078] In order to verify the accuracy of the identification method in the embodiment 1, the present embodiment also adopts the following tests to verify:

[0079] (1) On April 16, 2020, 200 saddle grouper broodstocks with a size of 123.15±24.56kg were selected and cultivated by Hainan Chenhai Aquaculture Co., Ltd. At this time, the disease resistance of the fry cannot be judged from the appearance, After clipping the fins and caudal fins, electronic markers were placed on the back muscles of each fish, and the corresponding marker number of each fish was recorded.

[0080] (2) Extract the DNA of each fish for resequencing to obtain SNP site information of each fish.

[0081] (4) According to the resequencing results, the Genome Association and Prediction Integrated Tool (GAPIT3) was used to calculate the breeding value. The number of fish with breeding value greater than 28 was 32, and the number of small fish 28 was 168 (Table 3).

[0082] (5) 32 disease-resistant parents a...

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Abstract

The invention discloses a method for selecting nervous necrosis virus resistant individuals of epinephelus lanceolatus. The method comprises the following steps: (1) obtaining SNP site information of the epinephelus lanceolatus; (2) analyzing the disease resistance weight value of the SNP marker and selecting the SNP marker; (3) obtaining candidate epinephelus lanceolatus individual SNP site information and selecting individuals with nervous necrosis virus resistance; and (4) detecting the selection accuracy of disease-resistant individuals. According to the method, the SNP marker with the maximum nervous necrosis virus resisting weight is screened from the SNP markers of the epinephelus lanceolatus, so that a method capable of selecting nervous necrosis virus resisting individuals is established.

Description

technical field [0001] The invention belongs to the technical field of grouper breeding, and in particular relates to a method for selecting an individual against nerve necrosis virus of saddle grouper. Background technique [0002] The saddle grouper, also known as the gentian grouper or the dragon grouper, belongs to the order Perciformes, the family Serranidae, and the genus Epinephelus. It is the largest of the grouper fish and is also known as the "Spot King". The saddle grouper is large in size, rich in nutrition, delicious in meat and fast in growth. It is a favorite marine fish treasure. The saddle grouper has become the main object of marine aquaculture in Guangdong, Hainan, Fujian, Guangxi and other places along the southern coast of China, with huge economic value. [0003] At present, with the continuous increase of the breeding volume, the problems of diseases and insect pests in the aquaculture of grouper are increasingly emerging, especially Viral nervous ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156C12Q2600/124Y02A40/81
Inventor 王庆秦启伟杨敏段旭琢梁凯珊
Owner SOUTH CHINA AGRI UNIV
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