Molecular marker combination for Litopenaeus vannamei germplasm identification and application thereof

A molecular marker, Vanabine technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of distinguishing different varieties, difficult to guarantee the authenticity of varieties and seedlings, disrupting the market operation order, etc. problem to improve the accuracy

Active Publication Date: 2016-08-17
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although different breeds have their own characteristics, it is difficult to distinguish different breeds from the phenotype
There are many and miscellaneous companies engaged in cultivating and raising seedlings of Litopenaeus vannamei in the market. Since the germplasm seedling certification system has not yet been implemented, it is difficult to guarantee the authenticity of the species and seedlings. , Farmers bring a lot of inconvenience, it is easy to replace the imported variety B with the imported variety A, or replace the selected variety with a non-selected variety in the market, which seriously disrupts the normal market operation order, and due to different varieties of seed There is no obvious difference at the seedling stage, but there is a huge difference in the later breeding process. Farmers will suffer a great loss of breeding benefits after obtaining inferior seedlings. Therefore, it is necessary to identify different Litopenaeus vannamei germplasm

Method used

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  • Molecular marker combination for Litopenaeus vannamei germplasm identification and application thereof
  • Molecular marker combination for Litopenaeus vannamei germplasm identification and application thereof
  • Molecular marker combination for Litopenaeus vannamei germplasm identification and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Development of a method for identification of Litopenaeus vannamei germplasm by combination of microsatellite markers

[0027] 1. Source of materials: Collection of germplasm materials from Zhengda, Kona Bay, SIS, Molokai, Ecuador, Kehai No. 1, and Guihai No. 1. Bay includes materials from 2012, 2014 and 2015, SIS includes materials from 2015 and 2016, materials from Guihai 1, Kehai 1, Molokai and Ecuador from 2014, the total number of samples 192.

[0028] 2. DNA extraction: The DNA of the above-mentioned materials was extracted using the Tiangen Plant Genome Extraction Kit. For the method, refer to the kit instructions. The concentration of the extracted DNA was measured using a Nanodrop 1000 (Thermo, USA), and stored in a -20°C refrigerator.

[0029] 3. Dilute the DNA extracted above to 50-100ng / μl respectively, use 13 pairs of high polymorphic microsatellite marker primers of the present invention to carry out PCR amplification, and the microsatellite pr...

Embodiment 2

[0037] Example 2: Application of the method for molecular identification of Litopenaeus vannamei germplasm

[0038] In view of the fact that the method established in the present invention has a high identification accuracy rate for different germplasms, the method of the present invention can be used to realize the identification of different germplasms of Litopenaeus vannamei and the species traceability of unknown source materials. The options offered are as follows:

[0039] 1. Collect germplasm materials from different domestic and international commercial germplasm companies. The number of individuals of each germplasm source is more than 30, and try to include materials from multiple generations and multiple regions. The DNA of the above-mentioned materials was extracted using the Tiangen Plant Genome Extraction Kit. For the method, refer to the kit instruction manual. The concentration of the extracted DNA was measured using a Nanodrop 1000 (Thermo, USA), and stored in...

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Abstract

The invention relates to a germplasm identification technology in the field of aquatic animal genetic breeding, in particular to a molecular marker combination for Litopenaeus vannamei germplasm identification and application thereof. The molecular marker combination is formed by 13 pairs of high-polymorphism microsatellite locus primers. A method for germplasm identification is established with the 13 pairs of high-polymorphism microsatellite markers, microsatellite typing is carried out on existing foreign imported germplasms including but not limited to the Zhengda breed, the SIS breed, the Kona Bay breed and the Molokai breed, and domestic artificially-selected germplasms including but not limited to the Kehai-1 breed and the Guihai-1 breed, molecular databases of different germplasm resources are established with obtained typing data, and different germplasms are identified with the molecular markers. The germplasm sources of Litopenaeus vannamei with unknown sources can be identified by comparing panel typing results with the established databases of different germplasm resources. The method for accurately identifying different germplasm materials of Litopenaeus vannamei with the molecular means has great significance on tracing, evaluation, protection and utilization of different germplasms.

Description

technical field [0001] The invention relates to germplasm identification technology in the field of genetic breeding of aquatic animals, in particular to a combination of molecular markers for germplasm identification of Litopenaeus vannamei and its application. Background technique [0002] my country's Litopenaeus vannamei was first introduced from Hawaii, USA in 1988 by Zhang Weiquan, a researcher at the Institute of Oceanology, Chinese Academy of Sciences. Since this species is suitable for seawater farming and can be cultured in inland freshwater areas after desalination, this species has become my country's The prawn species with the largest farming scale has an annual output of more than 1 million tons in recent years, with an annual output value of more than 50 billion yuan. [0003] Farming puts seeds first, and improved seeds are also the basis for the healthy development of the entire Litopenaeus vannamei industry. Since Litopenaeus vannamei is an imported species...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 于洋相建海李富花王全超张晓军
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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