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Molecular marker for identifying genetic sex of haliotis discus hannai and application thereof

A kind of wrinkled abalone, molecular marker technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. problems such as decline, to achieve the effect of increasing economic benefits and improving breeding efficiency

Inactive Publication Date: 2020-10-09
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Especially in the breeding season, the gonads of aquaculture abalone mature and discharge one after another, not only wasting a lot of energy in vain, resulting in a decline in the quality and st

Method used

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  • Molecular marker for identifying genetic sex of haliotis discus hannai and application thereof
  • Molecular marker for identifying genetic sex of haliotis discus hannai and application thereof
  • Molecular marker for identifying genetic sex of haliotis discus hannai and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Specific molecular markers of abalone wrinkled

[0032] The applicant of the present invention took the wrinkled plate abalone collected in Jinjiang, Fujian, China, and the Jinjiang Fuda abalone farm as samples, and constructed 11 sets of genome resequencing data of the wrinkled plate abalone, each of male and female, and performed GWAS and Sexually dimorphic genotype analysis, get the genome-wide association analysis result map and sex dimorphic SNP density distribution map (see figure 1 ). It can be seen from the figure that the significantly related regions of sex are located on chromosomes 2 and 9, and the most significantly related sites on chromosome 9 are concentrated in the terminal region of 9. Through the indel detection of sexual dimorphism on chromosome 9, a sequence was finally found in the sex-significantly related region of chromosome 9 that was specifically deleted in male Abalone rugosa, and the sequence that was finally screened for the spe...

Embodiment 2

[0035] Example 2: Sex identification of wrinkled pan abalone

[0036] Source of wrinkled plate abalone: ​​58 from Jinjiang population, 59 from Dalian population, and 55 from South Korea population.

[0037] The primer pairs of the present invention are used to carry out PCR amplification on Genomic DNA of Panbao wrinkle, which is determined to be male or female by anatomical examination, and the accuracy and specificity of the method of the present invention are verified by comparing the agarose gel electrophoresis patterns. The specific method steps are as follows:

[0038] 1. Cut out 12 parts of the adductor muscle tissue of male and 12 female wrinkled abalone adults, put them in absolute ethanol, and store them at -20°C for genome extraction;

[0039] 2. Genomic DNA was extracted by the phenol-chloroform method, and the obtained genomic DNA was uniformly diluted to 30ng / uL as a template for later use;

[0040] 3. Use primer MSP-F / R for PCR amplification:

[0041] (1) PCR a...

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Abstract

The invention discloses a molecular marker for identifying genetic sex of haliotis discus hannai and an application of the molecular marker. The molecular marker is shown as insertion/deletion lengthpolymorphism of a nucleotide sequence shown as SEQ ID NO: 1; an individual with the nucleotide sequence as shown in SEQ ID NO: 1 is expressed as genetic male haliotis discus hannai. During detection,SEQ ID NO: 2 and SEQ ID NO: 3 are adopted as primers for PCR amplification, and when the amplified fragments are 252bp and 240bp, the haliotis discus hannai to-be-detected lacks the gene of the nucleotide sequence shown in SEQ ID NO: 1 and is genetic male haliotis discus hannai; when only 252bp amplified fragments exist, the haliotis discus hannai to-be-detected has a nucleotide sequence shown asSEQ ID NO: 1 and is genetic female haliotis discus hannai. The molecular marker disclosed by the invention can be used for simply, conveniently, quickly and stably identifying the genetic sex of the haliotis discus hannai, is beneficial to developing a unisexual breeding technology of the haliotis discus hannai and developing haliotis discus hannai breeding, further improves the breeding benefit and increases the economic benefit of haliotis discus hannai breeding.

Description

technical field [0001] The invention relates to the technical field of aquatic organisms, in particular to a pair of primers and a method for identifying the sex of abalone rugosa. Background technique [0002] Gender is a key link in determining the reproduction of bisexual organisms, and it is also a continuous hotspot in the research of developmental biology and evolutionary biology. Shellfish (molluscs) are closely related to humans. They may be an important protein source for humans, or they may be intermediate hosts for pathogens or invasive organisms that endanger the environment. Shellfish is the second door of the animal kingdom, with a wide range of species, rich and diverse reproductive modes and sex determination methods, which can provide rich materials for the study of the sex determination mechanism and overall evolution of invertebrates. Individual growth and other important economic traits of some shellfish species are closely related to sex; however, the s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6879C12Q1/6858C12Q2600/156
Inventor 蔡明夷罗晗娇
Owner JIMEI UNIV
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