Low-salt-tolerant molecular marker C62 for portunus trituberculatus and application of low-salt-tolerant molecular marker C62

A technology of Portunus trituratus and molecular markers, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems affecting the normal molting and growth of adult crabs, feeding rate, metamorphosis rate and survival rate of juvenile crabs decline, lack of markers for molecular marker-assisted breeding, etc., to achieve the effect of promoting healthy reproduction, increasing feeding rate, and the method is accurate and reliable

Active Publication Date: 2020-02-04
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
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Problems solved by technology

Portunus trituberculatus adapts to a salinity of 20-35. However, because it is mainly cultured in outdoor ponds or tidal flats, the salinity of the cultured water often drops sharply due to factors such as heavy rains, which makes the feeding rate, metamorphosis rate and survival rate of young crabs Significantly decreased, it will also affect the normal molting and growth of adult crabs
At present, there are few studies on the development of low-salt tolerance molecular markers in Portunus trituberculatus, and the industry lacks markers that can be applied to molecular marker-assisted breeding

Method used

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  • Low-salt-tolerant molecular marker C62 for portunus trituberculatus and application of low-salt-tolerant molecular marker C62
  • Low-salt-tolerant molecular marker C62 for portunus trituberculatus and application of low-salt-tolerant molecular marker C62
  • Low-salt-tolerant molecular marker C62 for portunus trituberculatus and application of low-salt-tolerant molecular marker C62

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Embodiment 1

[0022] 1. Screening of candidate molecular markers related to low-salt tolerance traits

[0023] 1. Sequencing data filtering and comparison

[0024] The DNA extraction was carried out using the kit of Quanshijin Company, and the DNA extraction was carried out by using the principle of specific adsorption of DNA on the silica gel membrane spin column. First, put about 30mg of tissue samples into a 1.5ml sterile enzyme centrifuge tube, add 200μl LysisBuffer 8 (LB8) and 20μl RNaseA (10mg / ml), shake for about 10s, incubate at room temperature for 2min, add 20μl Proteinase K (20mg / ml), fully shake and mix, incubate at 55°C until completely lysed, add 1.5 times the volume of Binding Buffer 8 (BB8), mix well and add to the spin column, in a high-speed low-temperature refrigerated centrifuge (Eppendorf5804R) Centrifuge at 12000rpm for 30s, and discard the waste liquid. Then add 500μl of Clean Buffer 8 (CB8), centrifuge at 12000rpm for 30s, discard the waste solution (repeat once), ...

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Abstract

The invention provides a low-salt-tolerant molecular marker C62 for portunus trituberculatus and application of the low-salt-tolerant molecular marker C62. A nucleotide sequence of the molecular marker C62 is as shown in SEQ ID No.1, and nucleotide sequences of a primer pair for detecting the molecular marker C62 are as shown in SEQ ID No.2 and SEQ ID No.3. The molecular marker C62 is an SNP marker, and the low-salt tolerance genotype of the molecular marker C62 is an AG genotype. The molecular marker C62 provided by the invention is not limited by the growth stage of the portunus trituberculatus, is used for accelerating the breeding process of the portunus trituberculatus and quickly breeding crab species with excellent low-salt resistance, and is beneficial to healthy breeding and sustainable development of the portunus trituberculatus, so that the molecular marker C62 has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of DNA molecular markers, and in particular relates to a low-salt-tolerant molecular marker C62 of Portunus trituberculatus and an application thereof. Background technique [0002] Portunus trituberculatus (Portunus trituberculatus) belongs to Crustacea, Decapoda, Portunidae, Portunidae, commonly known as Portunus, and is an important large-scale marine economic crab in my country. Swimming crab meat is delicious and nutritious. It enjoys a high reputation at home and abroad and is deeply loved by consumers. Portunus trituberculatus adapts to a salinity of 20-35. However, because it is mainly cultured in outdoor ponds or tidal flats, the salinity of the cultured water often drops sharply due to factors such as heavy rains, which makes the feeding rate, metamorphosis rate and survival rate of young crabs Significantly decreased, it will also affect the normal molting and growth of adult crabs. Therefore, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 吕建建张文赵笑颜刘萍李健
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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