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Application of SNP sites at 203 and 752 in hypoxia inducible factor as molecular markers in hypoxia-resistant breeding of megalobrama amblycephala

A technology of hypoxia-inducible factors and molecular markers, which is applied in the field of breeding to achieve high hypoxia tolerance and speed up the breeding process

Pending Publication Date: 2022-02-18
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Indirectly regulate the body's ability to adapt to hypoxia by regulating the HIF family genes, but the regulation is controlled by a variety of factors (O 2 , α-ketoglutarate, Fe 2+ , ascorbate, etc.) restrictions

Method used

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  • Application of SNP sites at 203 and 752 in hypoxia inducible factor as molecular markers in hypoxia-resistant breeding of megalobrama amblycephala
  • Application of SNP sites at 203 and 752 in hypoxia inducible factor as molecular markers in hypoxia-resistant breeding of megalobrama amblycephala
  • Application of SNP sites at 203 and 752 in hypoxia inducible factor as molecular markers in hypoxia-resistant breeding of megalobrama amblycephala

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Experimental program
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Effect test

Embodiment 1

[0025] Materials and methods

[0026] 1.1. Experimental fish

[0027] The samples of bream were obtained from the Bream Genetics and Breeding Center (BGBC) of Shanghai Ocean University. This application selects "Pujiang No. 2" low-oxygen-tolerant variety and wild pomace bream in Poyang Lake as samples. "Pujiang No. 2" is a new low-oxygen-tolerant strain based on the wild pomace bream in Poyang Lake, which was bred through 4 generations of continuous breeding by adopting group breeding technology supplemented by low-oxygen stress technology. Juvenile fish (30±5g) were domesticated for 2 weeks in an indoor tank (25±0.1°C, dissolved oxygen concentration in water was 8.5±0.5mg·L -1 ).

[0028] 1.2. Hypoxic experiment

[0029] "Pujiang No. 2" low-oxygen resistant strain and juvenile wild bream, 36 in each group, were placed in two 25L glass tanks for 2 days. Tanks were filled with dechlorinated tap water sparged with air. Dissolved oxygen content in water was monitored using ...

Embodiment 2

[0048] 2.1. Analysis of SNP loci of HIFα gene in bream

[0049] Two linkage disequilibrium SNPs at exons 203 and 752 of the HIF2αb gene with high genetic diversity (PIC>0.5) were identified by analyzing the transcriptome data of "Pujiang 2" low-oxygen tolerant cultivar bream site (Table 2). Two SNP sites are non-synonymous mutations, T 203 A 203 The codon change type of the site is AAA-ATA, which code for lysine and isoleucine located in the PAS-A domain, respectively. C 752 A 752 The codon change type of the site is TAT-TCT, encoding tyrosine and serine respectively located in the PAS-B domain. There are two haplotypes, haplotype I (HI, T 203 C 752 ) and haplotype Ⅱ (HⅡ, A 203 A 752 )(table 3). Haplotype I mainly appeared in wild bream, and haplotype II mainly appeared in "Pujiang No. 2" low-oxygen-tolerant variety (Table 3). Two haplotypes randomly form three diplotypes, including homozygous diplotype I (DI, T 203 T 203 C 752 C 752 ), homozygous diplotype II (...

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Abstract

The invention provides application of SNP sites at 203 and 752 in a hypoxia inducible factor as molecular markers hypoxia-resistant breeding of megalobrama amblycephala; according to application of haplotypes of the SNP sites in megalobrama amblycephala hypoxia-resistant breeding, the haplotypes comprise a haplotype I and a haplotype II, the haplotypes are combined to obtain a double type. The double type comprises a homozygous double I type, a homozygous double II type and a heterozygous double III type, two SNP sites of the megalobrama amblycephala hypoxia inducible factor gene and the megalobrama amblycephala hypoxia-resistant character are subjected to correlation analysis, and it can be known that megalobrama amblycephala LOEcrit value, red blood cell number, hypoxia-resistant related enzyme activity and Epo gene expression have significant differences in hypoxia-resistant genotype individuals; therefore, the molecular marker can be used as a molecular marker related to the hypoxia-resistant character so as to accelerate the breeding process of megalobrama amblycephala hypoxia-resistant improved variety, that is, a double type II individual shows relatively high hypoxia-resistant capability, and a haploid type II can be used for molecular assisted breeding of a megalobrama amblycephala hypoxia-resistant new strain.

Description

technical field [0001] The invention belongs to the technical field of breeding, and in particular relates to the application of 203 and 752 SNP sites in a hypoxia-inducible factor as molecular markers in hypoxia-resistant breeding of bream. Background technique [0002] Bream is native to the lakes attached to the Yangtze River, and is a herbivorous freshwater fish species with high economic value and strong disease resistance in my country. In 2019, the output of grouphead bream exceeded 7.6×10 5 Ton. However, bream is a species that is sensitive to low oxygen. When the temperature, weather and water quality change greatly, the dissolved oxygen concentration in the pond will decrease, so there will be high mortality in the process of breeding bream. Therefore, there is an urgent need to breed improved varieties of bream tolerant to hypoxia. [0003] In the application of the existing haplotypes of the SNP loci associated with hypoxia tolerance in bream, the haplotype of...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 邹曙明赵姗姗郑国栋陈杰
Owner SHANGHAI OCEAN UNIV