A Causal Mutation Site for Chicken Feather Turning Character and Its Application in Genetic Breeding

A mutation site, genetic breeding technology, applied to the causal mutation site of chicken feather traits and its application in genetic breeding, can solve the problems of incomplete genome marker information, cumbersome, time-consuming, etc., to increase steps and difficulty , Accelerate the breeding process and shorten the breeding cycle

Active Publication Date: 2022-04-29
JIANGXI AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The research method is simple and rough, and the gene mutations found in the research results are only related, not the causal mutation sites of feather turning, so the application of molecular breeding for later turning feather cannot achieve accurate breeding effect;
[0006] (2) The existing patent "Interval positioning method and application of the main gene of the chicken feather trait" (patent number: CN106967832 A) by constructing the F2 generation resource population with the trait of curl feather, and then using the simplified genome sequencing method and the whole genome association Analysis to locate the target interval, and finally use the candidate gene method to screen the main effect gene. The implementation of the entire identification plan takes a long time (the construction of the experimental population is generally at least two years), and the technical processing procedures are many and cumbersome. Genomic marker information is incomplete, which can only be used for marker linkage mapping or association mapping analysis, and cannot directly identify causal genes and causal mutation sites, and cannot quickly and accurately locate target genes and mutation sites
[0007] Based on the above content, it can be seen that there is no causal gene and its causal mutation that can quickly and accurately locate the causal gene that affects the feather turning of Xiushui yellow-feathered black-bone chicken in the prior art

Method used

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  • A Causal Mutation Site for Chicken Feather Turning Character and Its Application in Genetic Breeding
  • A Causal Mutation Site for Chicken Feather Turning Character and Its Application in Genetic Breeding
  • A Causal Mutation Site for Chicken Feather Turning Character and Its Application in Genetic Breeding

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Embodiment 1

[0047] Example 1 The method of identifying causal mutation sites of chicken feather turning traits based on whole genome deep resequencing

[0048] 1. Preparation of test samples

[0049] In this technical scheme, the collection of test samples is convenient and the preparation time is short, which can be mainly divided into two parts:

[0050] (1) Whole-genome deep resequencing samples: including purebred individuals of Xiushui yellow-feather black-bone chicken and samples of yellow chicken breeds from other places in Jiangxi that have a close genetic relationship with Xiushui yellow-feather black-bone chicken. The specific number of samples is: Xiushui yellow-feather black-bone chicken There were 9 flat-feathered black-bone chickens and 6 turned-feathered individuals; 12 Ningdu yellow chickens, which were closely related to Xiushui yellow-feathered black-bone chickens; 15 Guangfeng white-eared yellow chickens; the above samples were divided into feather-turned groups (test ...

Embodiment 2

[0074] Example 2 Identification of causal mutation sites for feather turning traits in chickens based on deep genome-wide resequencing

[0075] (1) Whole-genome deep resequencing of 42 tested chickens

[0076] After whole genome resequencing of 42 experimental chickens, it was found that the sequencing depth was 12.26×-22.47×, and the 1× whole genome coverage rate exceeded 97%, fully meeting the requirements of the present invention. A total of 872.94Gb of raw reads (raw reads) were obtained by sequencing, and 871.42Gb of clean data were obtained. The relevant data has been submitted to the public database GSAdatabase (CRA003681). After variation searching and filtering, a variation dataset of 13,110,337 SNPs and 1,075,314 indels was finally obtained.

[0077] (2) GWAS results of Xiushui yellow-feathered black-bone chicken

[0078] A univariate case-control model was used to carry out the association analysis of feather turning traits in Xiushui yellow-feathered black-bone c...

Embodiment 3

[0084] Application of embodiment 3 turning feather causal mutation

[0085] Using 95 Xiushui yellow-feathered black-bone chickens (50 of which were turned-feather individuals and 45 were flat-feathered individuals) as a large group of verification samples, the direct sequencing method of PCR products (see Table 2 for primers) was used to accurately detect the above two candidate causal factors respectively. Frequency distribution of mutation sites.

[0086] Table 2. Primer information for PCR detection of two candidate causal mutation sites

[0087]

[0088] The PCR reaction uses a 25 μL system: 10 × Taq DNA polymerase buffer (500mmol / L KCl, 100mmol / L Tris Cl, 15mmol / L MgCl 2 ) 2.5 μL, dNTPs (ATP, TTP, GTP, CTP concentration is 2.5mmol / L 1μL), primer (0.5μM) 1μL, Taq DNA polymerase (2.5U / μL) 0.25μL, genomic DNA template (20ng / μL) 40ng, ddH 2 O supplemented to 25 μL.

[0089] The amplification program was: 95°C for 3min, 94°C for 30s, 59°C for 35s, 72°C for 30s, 33 cycl...

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Abstract

The invention discloses a causal mutation site of chicken feather turning trait and its application in genetic breeding, belonging to the field of molecular genetic breeding. The invention discloses the application of a causal mutation site for chicken feather turning traits in genetic breeding. The causal mutation site for chicken feather turning traits is located in the 5.24-5.28M region of chicken No. 33 chromosome, and there is a deletion mutation g.5285437- 5285451delGATGCCGGCAGGACG, the chicken with the deletion mutation is a black-bone chicken. Based on genome deep resequencing, the present invention can quickly and accurately locate the causal gene and its causal mutation that affect feather turning of black-bone chickens in one step, and use the causal mutation site of feather turning as a molecular marker to realize rapid breeding of black-bone chicken populations.

Description

technical field [0001] The invention relates to the field of molecular genetic breeding, in particular to a causal mutation site for chicken feather turning and its application in genetic breeding. Background technique [0002] Xiushui yellow-feathered black-bone chicken is a newly discovered local chicken group in Xiushui County, Jiangxi Province. It is said that it has been widely raised in the local area during the late Ming and early Qing Dynasties. Xiushui yellow-feather black-bone chicken is a small-sized meat-type yellow-feather black-bone chicken with medicinal functions. It is suitable for wild stocking. It has the characteristics of small size, strong wildness, wide adaptability and resistance to rough feeding. Xiushui yellow-feathered black-bone chicken has both flat-feathered individuals and rolled-feathered (curly-feathered) individuals. [0003] Black-bone chicken with feathers has always been used as a medicinal record in Chinese traditional medicine. As reco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6888C12Q1/6869C12Q2600/156C12Q2600/124C12Q2535/122
Inventor 毛辉荣刘三凤席苏望陈彪周明芳胡晓龙付戴波吴康琦郭志豪刘水兵张文韬于士蒙齐振平王秋连
Owner JIANGXI AGRICULTURAL UNIVERSITY
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