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Genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of genetic molecular marking combination

A technology of molecular markers and genetic markers, which is applied in the direction of resistance to vector-borne diseases, recombinant DNA technology, and microbial measurement/inspection, can solve the problem of late development of sea cucumber selection and breeding, and achieve improved breeding survival rate and breeding Benefits, automatic procedural detection, clear target effect

Active Publication Date: 2018-06-15
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Compared with other economic aquatic organisms, the selection and breeding of improved species of sea cucumber was carried out relatively late, and the current breeding methods are still mainly based on selective breeding and hybrid breeding. Research on other core breeding technologies such as molecular assisted breeding and genome selective breeding is still in its infancy. stage

Method used

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  • Genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of genetic molecular marking combination
  • Genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of genetic molecular marking combination
  • Genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of genetic molecular marking combination

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Embodiment 1

[0023] Example 1: Acquisition of genetic molecular markers for breeding of disease-resistant sea cucumbers and their verification in the offspring of the family

[0024] A japonicus family was constructed, and the disease resistance of each family was evaluated through the challenge test. A japonicus family with moderate disease resistance was selected, and the 2 parents and 142 offspring of the family were used as the mapping group. With the help of a high-throughput sequencing platform , developed SLAF markers, and constructed a high-density genetic linkage map of A. japonicus 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 poly...

Embodiment 2

[0029] Example 2: Verification of Apostichopus japonicus disease resistance-related SNP molecular markers in an expanded population

[0030]According to the flanking sequences on both sides of the three SNP sites obtained by high-throughput sequencing, primers for SNP site amplification detection were designed, and the corresponding primer sequences were SNP88F 5'-TTTTGTCCCCAGAGCATTATCCAA-3' and SNP88R 5'-TCCAGGAATGATATTTAGTGGTTGT-3' ; SNP112F 5'-ATTTTGGTTAGGGTTAAGAAGGCT-3' and SNP112R 5'-GTTTCCAAACAACATATGCACTCC-3'; SNP126F 5'-GGATTGATCGGGGACACACA-3' and SNP126R 5'-TGGGCTAAGTCACCAATCTGC-3'. The three SNP loci related to disease resistance of A. japonicus were typed by using the HRM small fragment method in the expanded population by using the corresponding sequences, and the QTL loci were verified in combination with the related trait data of the expanded population. The expanded population used was 8-month-old seedlings cultivated in the same batch of large-scale seedlings i...

Embodiment 3

[0040] Example 3: Application of Genetic Molecular Marker Combination in the Breeding of Disease-resistant Apostichopus japonicus in the Production of Apostichopus japonicus

[0041] During the spring seedling period of A. japonicus, 100 parent references were randomly selected from the cultivated parent references, and the genotypes of 100 reference references at SNP88, SNP112, and SNP126 loci were genotyped using HRM technology. The primer sequences used were SNP88F 5′-TTTTGTCCCAGAGCATTATCCAA-3′ and SNP88R 5′-TCCAGGAATGATATTTAGTGGTTGT-3′; SNP112F 5′-ATTTTGGTTAGGGTTAAGAAGGCT-3′ and SNP112R 5′-GTTTCCAAACAACATATGCACTCC-3′ and SNP126FCATCG5′-GGGA '-TGGGCTAAGTCACCAATCTGC-3'. These three genetic marker amplification detection systems are: 1 μL (50 ng / μL) of DNA template, 4.5 μL 2×ES Taq Master Mix, 0.5 μL (10 μmol / L) of upstream and downstream primers, 3.5 μL ddH 2 O; PCR reaction program for 3 genetic marker typing: pre-denaturation at 95°C for 5 min; denaturation at 94°C for 30...

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Abstract

The invention discloses a genetic molecular marking combination for anti-disease stichopus japonicas breeding and application of the genetic molecular marking combination and belongs to the field of aquatic product genetic breeding. The genetic molecular marking combination consists of three SNP (Single Nucleotide Polymorphism) markers SNP88, SNP112 and SNP126 related to anti-vibrio splendidus infection of stichopus japonicas, and the three marked dominant genotypes are respectively SNP88 (CC), SNP112 (AA) and SNP126 (TT). A breeding method with the genetic markers comprises the following operation steps: (1) extracting parent DNA (Deoxyribonucleic Acid) and typing; (2) selecting parents; (3) performing filial generation seedling breeding. By adopting the genetic molecular marking combination, effective molecular markers for improving the economic property of anti-vibrio splendidus infection of stichopus japonicas are provided, the genetic markers are adopted for auxiliary marker selection of stichopus japonicas, anti-disease breeding can be carried out, the anti-vibrio splendidus infection of the stichopus japonicas can be improved, the morbidity of skin ulcer syndromes of the stichopus japonicas can be substantially reduced, and the combination has great significances in increasing the breeding survival rate and the breeding benefits.

Description

technical field [0001] The invention belongs to the field of aquatic genetics and breeding, in particular to a combination of genetic molecular markers for disease-resistant sea cucumber selection 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 changes, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156Y02A50/30
Inventor 廖梅杰王印庚李彬刘安然荣小军陈贵平范瑞用张正
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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