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A molecular marker and its application for assisted selection breeding of sea cucumber growth traits

A technology of growth traits and molecular markers, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of feasibility and accuracy that need to be further verified, and achieve high accuracy and selection Efficient, targeted results

Active Publication Date: 2018-10-30
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

However, these markers have not undergone further expansion verification and production verification, and the feasibility and accuracy of their application need to be further verified

Method used

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  • A molecular marker and its application for assisted selection breeding of sea cucumber growth traits
  • A molecular marker and its application for assisted selection breeding of sea cucumber growth traits
  • A molecular marker and its application for assisted selection breeding of sea cucumber growth traits

Examples

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Effect test

Embodiment 1

[0021] Example 1: Acquisition of genetic molecular markers for breeding of sea cucumber growth traits and their verification in the offspring of the family

[0022] A sea cucumber family was constructed, and the 2 parents and 142 offspring of the family were used as the mapping population. With the help of a high-throughput sequencing platform, SLAF markers were developed, and a high-density genetic linkage map of sea cucumber was constructed using HighMap software. After high-throughput sequencing, a total of 264,810 SLAF tags were developed, including 112,322 polymorphic SLAF tags. The average sequencing depth of parents with SLAF tags was 23.67×, and the average sequencing depth of offspring was 5.44×. Through bioinformatics analysis, there are 50,905 tags that can be used for genetic map construction. After quality filtering, 4629 polymorphic tags with high sequencing depth, high integrity and no partial separation were obtained. The screened 4629 SLAF tags were used for ...

Embodiment 2

[0026] Example 2: Verification of Apostichopus japonicus growth-related SNP molecular markers in an expanded population

[0027] According to the flanking sequences on both sides of the SNP40 site obtained by high-throughput sequencing, primers for SNP site amplification detection were designed, and the corresponding primer sequences were SNP40F: 5'-GGGTGGACTTGGAGCAGAAG-3' and SNP40R: 5'-ACCAACGTCCTCTCTGGTTATCA-3' . The corresponding sequence was used to type the SNP site in the expanded population by using the HRM small fragment method, and the QTL site verification was carried out in combination with the related trait data of the expanded population. The expanded population used was 10-month-old seedlings from the same batch of large-scale breeding and cultured in the same breeding environment. 96 individuals were randomly selected, and the body weight of the individuals was measured. Use the HRM method to detect individual genotypes. The marker amplification detection syst...

Embodiment 3

[0031] Example 3: Application of molecular markers for growth traits-assisted selection breeding in Apostichopus japonicus breeding and production

[0032]During the spring seedling stage of A. japonicus, 100 parent references were randomly selected from the cultivated parent references, and the genotypes of the 100 parent references at the SNP40 locus were genotyped using HRM technology. The primer sequences used were SNP40F: 5'-GGGTGGACTTGGAGCAGAAG-3' and SNP40R: 5'-ACCAACGTCCTCTCTGGTTATCA-3', and the amplification detection system was: DNA template 1 μL (50ng / μL), 4.5 μL 2×ES Taq Master Mix, 0.5 μL (10 μmol / L) each of upstream and downstream primers, 3.5 μL ddH 2 O; PCR reaction program: pre-denaturation at 95°C for 5 minutes; denaturation at 94°C for 30 s, annealing at 60.5°C for 30 s, extension at 72°C for 30 s, a total of 35 cycles; final extension at 72°C for 7 min, and storage at 4°C; markers were typed using high-resolution Melting curve analysis technique (HRM) meth...

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Abstract

The invention discloses a molecular marker for stichopus japonicus growth property auxiliary selective breeding and application thereof, and belongs to the field of molecular marker auxiliary breeding. The molecular marker is an SNP40 marker; a primer pair of the marker comprises SNP40F:5'-GGGTGGACTTGGAGCAGAAG-3' and SNP40R:5'-ACCAACGTCCTCTCTGGTTATCA-3'; the marker is used for growing the propertyauxiliary breeding advantage genotype of CC. Meanwhile, the invention also discloses the auxiliary breeding steps of the marker. The effective molecular marker is provided for improving the economicproperty of the stichopus japonicus growth in a genetic aspect; the genetic marker is used for performing marker auxiliary selection on the stichopus japonicus; the genetic breeding is performed; thegrowth speed of stichopus japonicus seeds can be effectively accelerated; important significance is realized on improving the culture benefits.

Description

technical field [0001] The invention belongs to the field of molecular marker assisted breeding, and in particular relates to a molecular marker for assisted selection breeding of sea cucumber growth traits and its application. Background technique [0002] Sea cucumber belongs to Echinodermata phylum Echinodermata and sea cucumber class Holothuroidea, and is one of the important sea treasures in northern my country. In recent years, sea cucumber farming in my country has developed rapidly, and its scale has expanded rapidly. The direct economic output value is about 30 billion yuan. It is the species with the largest single output value in my country's mariculture industry. important way. However, with the continuous expansion of the scale of the sea cucumber industry, a series of bottleneck problems that restrict or potentially restrict the development of the industry, such as germplasm degradation, slow growth, long breeding cycle, poor ability to resist environmental cha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 廖梅杰李彬王印庚荣小军刘安然陈贵平范瑞用张正
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI