SNP molecular marker related to skin color character of black-bone chicken and application

A molecular marker and black-bone chicken technology, applied in the field of agricultural biology, can solve the problems of long test cross cycle, high cost, and difficulty in implementing the whole group, so as to improve the efficiency of seed selection and save the cost of breeding

Active Publication Date: 2016-05-25
HANGZHOU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is impossible to judge whether the genotype is homozygous based on the phenotype of the individual. Although it can be verified by test crossing with white-skinned chicken breeds, the test cross cycle is long and expensive, and it is difficult to implement it in the whole group

Method used

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  • SNP molecular marker related to skin color character of black-bone chicken and application
  • SNP molecular marker related to skin color character of black-bone chicken and application
  • SNP molecular marker related to skin color character of black-bone chicken and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 SNP molecular marker detection method of melanin major gene Fm

[0024] 1. Experimental equipment

[0025] Ultra-low temperature refrigerator (Thermo, USA); high-speed refrigerated centrifuge (Eppendorf5417R, Germany); gradient PCR instrument (ABIPCRsystem9700, USA); adjustable micropipette (Eppendorf, Germany); GELEQ electrophoresis gel imaging system (Bio-Rad, USA); UV-Vis spectrophotometer (BioSpec-nano, Japan).

[0026] 2. Genomic DNA extraction from chicken blood

[0027] Venous blood from chicken wings was collected and preserved in 75% ethanol. Genomic DNA from chicken blood was extracted with AxyPrep Total Blood Genomic DNA Mini Kit (Axygen, USA). After extraction, the DNA quality was detected by 1% agarose gel electrophoresis; the DNA concentration was measured by a UV-Vis spectrophotometer.

[0028] 3. Primer design

[0029] Specific primers were designed for the Fm gene segment of chromosome 20 of the Gallusgallus genome (NC_006107.3) in the NC...

Embodiment 2

[0039] Example 2 Verification test of skin color test cross of Dongxiang green-shell layer hens

[0040] 1. Test materials and methods

[0041] Dongxiang green-shell layer hen is a local silky breed native to Jiangxi Province. The skin color of the group is mainly black or light black, and a few individuals are white, indicating the existence of homozygosity and heterozygosity of the Fm gene. In the experiment, 23 roosters were randomly selected from Dongxiang green-shell hen population, and genomic DNA was extracted from wing vein blood, and the detection was carried out according to the steps in Example 1. According to the test results, a total of 12 Dongxiang roosters of three genotypes were selected, and they were tested with 72 Xing'an Ma chickens (local chicken breeds in Guangxi Province, white skin and black shank) at a breeding ratio of 1:6, and the eggs were hatched and hatched Afterwards, observe and record the skin color of the offspring.

[0042] 2. Results and A...

Embodiment 3

[0047] Application of embodiment 3 SNP marker detection method in different chicken species

[0048] 1. Experimental materials and methods

[0049] Select several Jiangshan silky chickens, Yisha B-series layer chickens, Yisa D-series layer chickens, "Dongxiang×Isa B" hybrid chickens, and "Dongxiang×Xing'an" hybrid chickens, and collect wing vein blood to extract genomic DNA. , detected according to the steps in Example 1. The SNP genotype of the samples was determined according to the results of enzyme digestion and electrophoresis, and the chi-square test was used for correlation analysis.

[0050] 2. Results and Analysis

[0051] The detection results of SNP markers of various chicken breeds are shown in Table 3.

[0052] Table 3 SNP typing results of different chicken breeds

[0053]

[0054] Jiangshan black-bone chicken is native to Jiangshan City, Zhejiang Province. It has flaky white feathers, black skin, black meat, and black bones. It is a reference chicken bree...

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Abstract

The invention provides application of an SNP molecular marker in detecting the skin color character of a black-bone chicken. The nucleotide sequence of the molecular marker is shown as SEQ ID No.1, and the base mutation type at the 171st bp is A / G. The invention further provides a primer sequence for detecting an SNP locus and a PCR-RFLP method. The genotype of the locus can be distinguished based on the difference of an electrophoretic band after DraI enzyme digestion, and then the homozygosis and heterozygosis of a sample Fm gene can be judged. If the 342 bp band appears, the sample Fm gene is a GG homozygosis-based gene; if the two bands of 171 bp and 342 bp appear, the sample Fm gene is an AG heterozygosis-based gene; if the 171 bp band appears, the sample Fm gene is an AA recessive-homozygosis-based gene. By means of the SNP molecular marker and the detection method, the individual genotype of the black-bone chicken can be judged in the chickling stage, early selection and early elimination of the skin color character can be achieved, and thus significant practical value can be achieved on improving selection efficiency and saving breeding cost.

Description

technical field [0001] The invention relates to a SNP marker used for detecting homozygous and heterozygous Fm genes related to skin color traits of black-bone chickens, and belongs to the field of agricultural biotechnology. Background technique [0002] Black-bone chicken has a very long history of breeding in my country. It usually has the characteristics of black skin, black bones, and black meat. It is a traditional and rare chicken species for both medicine and food in my country. It is mild, non-toxic, tonic for deficiency and fatigue, cures thirst, benefits puerpera, treats women's collapse and leukorrhea, and all diseases of deficiency and loss", so it is deeply favored by consumers. Silkie breeds mainly include Taihe Silky Feather Silky, Dongxiang Green-shell Layer, Jiangshan Silky, etc., all of which are commonly used as excellent materials in silky breeding. However, with the introduction of foreign chicken breeds and mixed blood, There are cases of incomplete bl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 王欢欢章学东张雷楼立峰陈美玲李庆海张成先陈贤惠
Owner HANGZHOU ACAD OF AGRI SCI
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