A detection reagent, method and application for identifying feather traits of chicken

Abstract

The application discloses a detection reagent, a method and application for identifying feather traits of chickens. It is found that when the feather phenotype of a chicken is curled feather, an InDel site exists at 2040246-2040259 bp of chromosome 34 of an Ensembl chicken reference genome GRCg7b version 34, the deleted nucleotide sequence is shown as SEQ ID NO:1, the site is a homozygous deletion type, and the genotype of a SNP site g.2040608G>A is AA type. The two sites are combined for use in early assisted selection of curled feather chickens. The two sites are detected by using the detection reagent, and the feather phenotype of the chicken can be quickly, effectively and accurately predicted, so that the breeding time is shortened and the breeding process is accelerated.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of poultry breeding, and in particular to a detection reagent, a method and an application for identifying feather traits of a chicken. BACKGROUND

[0002] Yuexi coiled-feather chicken, also known as Kylin chicken, is a high-quality local chicken breed originating from the western region of Guangdong Province, China, and has a history of over 840 years. The breed is named for its unique coiled feathers, with partial skin exposure, excellent heat dissipation performance, strong adaptability, and good growth performance. The unique coiled feather trait of the Yuexi coiled-feather chicken is beneficial for heat dissipation from the body surface, suitable for growth in the high-temperature and high-humidity climate conditions of the western region of Guangdong, and has strong stress resistance, which is of great significance for breeding heat-stress-resistant chicken breeds. In addition, as a local excellent breed, the Yuexi coiled-feather chicken is not only an important economic animal in the local area, but also a part of culture and diet, and is deeply loved by people. Protection and breeding work of the Yuexi coiled-feather chicken is being carried out to prevent loss of genetic resources. Therefore, it is necessary to establish a molecular technique capable of quickly and simply identifying the coiled feather trait.

[0003] SNP refers to a change in the base of a single nucleotide in a DNA sequence, and InDel refers to the insertion or deletion of one or more bases in a DNA sequence. These variations can affect the regulation mechanism of gene function and expression. InDel and SNP are the most common genetic variations in biology and the most commonly used molecular markers. They have a large number of markers, high coverage of whole genome, high diversity, and strong genetic stability, and are therefore widely used in marker-assisted selection in molecular breeding. However, there are still few related sites and identification techniques for the coiled feather trait of chicken, which cannot meet the needs of the breeding industry. SUMMARY

[0004] In order to overcome the above-mentioned defects and shortcomings in the prior art, the present application provides a detection reagent, a method and an application for identifying feather traits of a chicken.

[0005] A first object of the present application is to provide an application of a composition in identifying feather traits of a chicken.

[0006] A second object of the present application is to provide a detection reagent of the above-mentioned composition.

[0007] A third object of the present application is to provide a kit.

[0008] A fourth object of the present application is to provide an application of the above-mentioned detection reagent and / or kit in identifying feather traits of a chicken.

[0009] A fifth object of the present application is to provide an application of the above-mentioned detection reagent and / or kit in preparing a breeding product of a chicken.

[0010] A sixth object of the present application is to provide a method for identifying the feather trait of a chicken.

[0011] The present application claims the following:

[0012] Use of a composition in identifying the feather trait of a chicken, the composition being an InDel site and an SNP site;

[0013] The InDel site is located at 2040246-2040259 bp on chromosome 34 of the chicken genome, and the deleted nucleotide sequence is shown in SEQ ID NO: 1;

[0014] The SNP site is located at 2040608 on chromosome 34 of the chicken genome, and is a G or A polymorphism, which exists in GG and AA genotypes;

[0015] The chicken genome is the Ensembl database GRCg7b version;

[0016] The InDel site is homozygous deletion type, and the genotype of the SNP site is AA, and the feather phenotype of the individual is curled feather;

[0017] The InDel site is homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is flaky feather;

[0018] The InDel site is homozygous wild type or heterozygous type, and the feather phenotype of the individual is flaky feather.

[0019] The detection reagent of the above-mentioned composition is a primer with a nucleotide sequence as shown in SEQ ID NO: 2-3, and the detection reagent of the SNP site is a primer with a nucleotide sequence as shown in SEQ ID NO: 4-5.

[0020] A kit comprising the above-mentioned detection reagent.

[0021] Preferably, the kit further contains PCR amplification reagents and ddH2O.

[0022] Use of the above-mentioned detection reagent and / or kit in identifying the feather trait of a chicken.

[0023] Use of the above-mentioned detection reagent and / or kit in preparing a breeding product of a chicken.

[0024] A method for identifying the feather trait of a chicken, detecting the above-mentioned composition with the above-mentioned detection reagent.

[0025] As an implementable way, the method comprises the following steps:

[0026] S1. extracting genomic DNA of a sample to be tested;

[0027] S2. performing PCR amplification on the genomic DNA obtained in step S1 using the detection reagent described above, and then performing sequencing on the PCR amplification product to obtain sequencing data;

[0028] S3. analyzing the sequencing data obtained in step S2, and determining the feather trait of the chicken to be tested according to the composition described above.

[0029] Preferably, in step S1, the chicken is a Yuexi coiled-feather chicken, a dwarf coiled-feather chicken, a Qibannong chicken, a dwarf chicken, a Wenchang chicken, a Huaixiang chicken, a Qingyuan chicken, a mustache chicken, or a hybrid offspring of the Yuexi coiled-feather chicken, the dwarf coiled-feather chicken, the Qibannong chicken, the dwarf chicken, the Wenchang chicken, the Huaixiang chicken, the Qingyuan chicken, or the mustache chicken as a parent.

[0030] The InDel site is a homozygous deletion type, and the genotype of the SNP site is AA, and the feather phenotype of the chicken is coiled feather;

[0031] The InDel site is a homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the chicken is vane feather;

[0032] The InDel site is a homozygous wild type or a heterozygous type, and the feather phenotype of the chicken is vane feather.

[0033] As an implementable manner, in step S2, the sequencing is Sanger sequencing.

[0034] Compared with the prior art, the present application has the following beneficial effects:

[0035] The present application discloses a detection reagent, a method and an application for identifying the feather trait of a chicken. The present application finds that there is an InDel site at 2040246-2040259bp of chromosome 34 of Ensembl chicken reference genome GRCg7b version 34, the deleted nucleotide sequence is shown as SEQ ID NO: 1, when the InDel site is a homozygous wild type or a heterozygous type, the feather phenotype of the chicken is vane feather; when the InDel site is a homozygous deletion type, the genotype of the SNP site g.2040608G>A is AA type, and the feather phenotype of the individual is coiled feather, and the genotype of the SNP site g.2040608G>A is GG, and the feather phenotype of the individual is vane feather. The two sites can be used in early assisted selection of coiled-feather chickens in combination. Using the detection reagent of the present application to detect the two sites can quickly, effectively and accurately predict the feather phenotype of the chicken, thereby shortening the breeding time and accelerating the breeding process. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1Manhattan plot and QQ-plot of InDel loci associated with chicken feathering traits.

[0037] Figure 2 Sanger sequencing results of different genotypes at the site of 2040245bp on chromosome 34 of Ensembl chicken reference genome (GRCg7b version) (g.2040246_2040259delGACCCTGTGACACC); A: homozygous deletion type individual, B: homozygous wild type individual, C: heterozygous individual.

[0038] Figure 3 Sanger sequencing results of different genotypes at the site of 2040608bp on chromosome 34 of Ensembl chicken reference genome (GRCg7b version) (g.2040608G>A); A: AA type individual, B: GG type individual. DETAILED DESCRIPTION

[0039] The present application will be further described in conjunction with specific examples, but the examples do not limit the present application in any form. Unless otherwise specified, the reagents, methods and equipment used in the present application are conventional reagents, methods and equipment in the technical field.

[0040] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0041] Example 1 Whole genome resequencing

[0042] I. Experimental methods

[0043] 1. Preparation of DNA samples

[0044] The whole genome DNA of blood of Yuexi feathering chicken was extracted according to phenol-chloroform method, and the quality and concentration of the DNA were detected by Nanodrop One spectrophotometer. The A260 / 280 ratio was 1.8-2.0, and the A260 / 230 ratio was 1.7-1.9, which was qualified. The qualified DNA samples were uniformly diluted to 50 ng / μL.

[0045] 2. Whole genome resequencing

[0046] (1) DNA library construction and whole genome sequencing

[0047] The DNA sample was broken by using Covaris ultrasonic instrument, and the short DNA fragments meeting the length requirement were obtained by adjusting the breaking parameters. The broken sample was selected by magnetic beads to make the sample band concentrated at about 300-400 bp; the amount of DNA sample after purification was detected by Qubit fluorescence quantifier.

[0048] The reaction system is prepared, the double-stranded DNA ends are repaired, and an A base is added to the 3' end. The ligation reaction system is prepared, and the adapter is ligated to the DNA. The PCR reaction system is prepared, and the reaction program is set to amplify the ligation product. The amplified product is subjected to fragment screening using magnetic beads.

[0049] After the PCR product is denatured into a single strand, the circularization reaction system is prepared, and after thorough mixing, the reaction is carried out at an appropriate temperature for a certain period of time to obtain a single-stranded circular product. After digesting the linear DNA molecules that are not circularized, the final library is obtained. The single-stranded circular DNA molecules are subjected to rolling circle replication to form a DNA nanoball (DNB) containing more than 300 copies. The obtained DNBs are used in high-density DNA nanochip technology and added to the reticular small holes on the chip. Sequencing is performed by combined probe anchor polymerization technology.

[0050] (2) Data filtering

[0051] The raw data is processed and analyzed using the filtering software SOAPnuke developed by Huada. The filtering parameters of the SOAPnuke software are "-n 0.001-l 10-q 0.5--adaMR 0.25--polyX 50--minReadLen 150". The effective data is obtained.

[0052] (3) Obtain data

[0053] Rolling chicken data: The paired-end sequencing data is aligned to the Ensembl chicken reference genome (GRCg7b version). The alignment rate of the sequencing data is more than 99%, which can be used for subsequent analysis. Finally, a VCF file containing SNP and InDel data is obtained through variation detection, filtering and annotation, which is used for subsequent analysis.

[0054] Feathered chicken (normal feather) data: The VCF file containing SNP and InDel data obtained from the previous 60 seven hundred and one chicken resequencing data in the laboratory and the fastq raw data of 25 QianDongNan small Xiang chicken publicly available in the NCBI database is processed by the server platform in the laboratory to obtain the VCF file containing SNP and InDel data.

[0055] (4) Whole genome association analysis

[0056] The VCF file was converted into tped format using plink software, and the data was filtered using the commands "--maf 0.05 --geno 0.1". The emmax-kin-intel64 program in the emmax software package was used to calculate the kinship matrix. The emmax-intel64 program in the emmax software package was used to combine the phenotype data and the kinship matrix for GWAS analysis. Manhattan plot and QQplot were drawn for the GWAS results using R language.

[0057] II. Experimental results

[0058] The GWAS analysis results are shown in Table 1. Figure 1 As shown in Table 1, the GWAS results show that there are a large number of InDel sites on chromosome 34 that are significantly associated with the frizzle trait. Among them, the InDel site (g.2040246_2040259delGACCCTGTGACACC) at position 2040245bp on chromosome 34 of the chicken reference genome (GRCg7b version) in Ensembl is one of the most strongly associated sites, and the resequencing data shows that the frizzle chickens are all homozygous deletion type, while the above feathered chicken resequencing data at this site is homozygous wild type or heterozygous type, indicating that this molecular marker is extremely likely to assist the selection of the frizzle trait.

[0059] Example 2: Primer for amplifying InDel site

[0060] I. Experimental method

[0061] Using DNA samples of Yuexi frizzle chicken, short frizzle chicken, Qibainong chicken, short chicken, Wenchang chicken, Huaixiang chicken, Qingyuan chicken and Huqu chicken, PCR amplification primers covering the region of the InDel (g.2040246_2040259delGACCCTGTGACACC) site at position 2040245bp on chromosome 34 of the Ensembl chicken reference genome (GRCg7b version) significantly associated with the frizzle trait screened in Example 1 were designed, the primer sequences are shown in SEQ ID NO:2 and SEQ ID NO:3, and the amplification product size is 643bp.

[0062] F1: 5'-GTGCTCAGAAGTCCCCCTCAACCT-3' (SEQ ID NO:2);

[0063] R1: 5'-CCATCTCTCCACGCTCCTCT-3' (SEQ ID NO:3).

[0064] The DNA samples of frizzle chickens and feathered chickens were subjected to PCR verification using the PCR amplification primers, and the samples used for PCR verification are shown in Table 1.

[0065] Table 1 PCR verification of samples used

[0066] Variety Number (only) YueXi roll feather chicken 130 Dwarf roll feather chicken 30 Qibannong chicken 30 Dwarf chicken 30 Wenchang chicken 30 Huiseng chicken 30 Qingyuan chicken 30 Figure 2 30

[0067] PCR amplification system: 2x Es Taq MasterMix 25 μL, upstream primer (10 μmol / L) 2 μL, downstream primer (10 μmol / L) 2 μL, DNA template 200 ng, and add sterile double distilled water to 50 μL system.

[0068] PCR reaction program: 94°C pre-denaturation 2 min; 94°C denaturation 30 s, 61°C annealing 30 s, 72°C extension 30 s, a total of 32 cycles; 72°C extension 5 min.

[0069] The PCR amplification product was detected by agarose gel electrophoresis, and if the electrophoresis band was consistent with the size of the target band, the obtained PCR product was sent to the company for DNA sequence determination, and the forward and reverse reactions of the gene fragment were measured. The sequence results obtained were compared with the Ensembl genome sequence, and the corresponding gene deletion condition was obtained.

[0070] II. Experimental results

[0071] The sequencing results are shown in Figure 2 . Among them, Figure 2 A in indicates that the site is homozygous deletion, Figure 2 B in indicates that the site is homozygous wild type, Figure 3 C in indicates that the site is heterozygous. The sequencing statistical results are shown in Table 2, wherein all the chickens with the frizzle trait are homozygous deletion type; and almost all the chickens with the single comb trait are homozygous wild type and heterozygous type except for the Silky chicken.

[0072] Most of the Silky chickens are homozygous wild type and heterozygous type, but homozygous deletion type also appears. Therefore, another site needs to be screened out, and assisted selection is carried out for this case (between frizzle chickens and Silky chickens).

[0073] Table 2 InDel site genotyping statistical results

[0074]

[0075]

[0076] Example 3 Primer for amplifying SNP site

[0077] I. Experimental method

[0078] A SNP site (g.2040608G>A) was found under the InDel (g.2040246_2040259delGACCCTGTGACACC) site at 2040245 bp on chromosome 34 of the Ensembl chicken reference genome (GRCg7b version) screened in Example 1, which showed significant association with the curled feather trait in the GWAS results.

[0079] The DNA samples of Yuexi curled feather chicken, dwarf curled feather chicken, Qibainong chicken, dwarf chicken, Wenchang chicken, Huixiang chicken, Qingyuan chicken and Huqu chicken were used to design PCR amplification primers covering the region for the SNP site, the primer sequences were as shown in SEQ ID NO: 4 and SEQ ID NO: 5, and the size of the amplification product was 259 bp.

[0080] F2: 5'-GCCCTGAAAGATGCCAAATCCAAAC-3' (SEQ ID NO: 4);

[0081] R2: 5'-GGTGAACTGAGGCTTTGGACCC-3' (SEQ ID NO: 5).

[0082] The DNA samples of Yuexi curled feather chicken (curled feather chicken, 30) and Huixiang chicken (feather chicken, 30) were subjected to PCR verification using PCR amplification primers, and the PCR amplification system was as follows: 2x Es Taq MasterMix 25 μL, upstream primer (10 μmol / L) 2 μL, downstream primer (10 μmol / L) 2 μL, DNA template 200 ng, and sterile double distilled water to 50 μL system.

[0083] The PCR reaction program was as follows: 94°C pre-denaturation for 2 min; 94°C denaturation for 30 s, 61°C annealing for 30 s, 72°C extension for 30 s, a total of 32 cycles; and 72°C extension for 5 min.

[0084] The PCR amplification product was subjected to agarose gel electrophoresis detection, and if the electrophoresis band was consistent with the size of the target band, the obtained PCR product was sent to the company for DNA sequence determination, and the forward and reverse reactions of the gene fragment were determined. The sequence results obtained were compared with the Ensembl genome sequence to obtain the corresponding gene mutation.

[0085] II. Experimental results

[0086] The sequencing results are shown in Table 4. Figure 3 Figure 3 A in the above table indicates that the site is AA genotype, Variety B in the above table indicates that the site is GG genotype. The sequencing statistical results are shown in Table 4, wherein the curled feather chicken is all AA genotype, and the Huixiang chicken is all GG genotype.​

[0087] Therefore, the primers for amplifying the SNP site (g.2040608G>A) can be used to distinguish the coiled-feather chicken and the bantam chicken. Therefore, the primers for amplifying the InDel site and the primers for amplifying the SNP site (g.2040608G>A) can be used in combination to accurately identify the feather traits of the chicken breeds selected in the present application, and the success rate and accuracy can reach 100%, which can be completely used in the actual identification of the chicken feather traits.

[0088] Table 3: Genotyping statistics of the SNP site (g.2040608G>A)

[0089] Sanger sequencing number GG AA YueXi roll feather chicken Huiseng chicken 30 0 30 ​ 30 30 0

[0090] Example 4: A method for identifying the feather traits of a chicken

[0091] 1. Extract the DNA of the chicken sample to be tested.

[0092] 2. Perform PCR amplification using the DNA of the chicken sample to be tested obtained in step 1 as a template:

[0093] (1) Amplify the InDel site (g.2040246_2040259delGACCCTGTGACACC)

[0094] The PCR reaction system is: 2x Es Taq MasterMix 25 μL, 2 μL of each primer with the nucleotide sequence shown in SEQ ID NO: 2-3, 200 ng of DNA template, and sterilized double distilled water to 50 μL system.

[0095] The PCR reaction program is: 94°C pre-denaturation for 2 min; 94°C denaturation for 30 s, 61°C annealing for 30 s, 72°C extension for 30 s, a total of 32 cycles; 72°C extension for 5 min.

[0096] (2) Amplify the SNP site (g.2040608G>A)

[0097] The PCR reaction system is: 2x Es Taq MasterMix 25 μL, 2 μL of each primer with the nucleotide sequence shown in SEQ ID NO: 4-5, 200 ng of DNA template, and sterilized double distilled water to 50 μL system.

[0098] The PCR reaction program is: 94°C pre-denaturation for 2 min; 94°C denaturation for 30 s, 61°C annealing for 30 s, 72°C extension for 30 s, a total of 32 cycles; 72°C extension for 5 min.

[0099] 3. The PCR amplification product obtained in step 2 is detected by agarose gel electrophoresis, if the electrophoresis band is consistent with the size of the target band and clear and bright, the PCR amplification product obtained in step 2 is sequenced, the sequencing result is compared with the chromosome 34 of Ensembl chicken reference genome (GRCg7b version) and genotyped.

[0100] 4. Result judgment

[0101] There is an InDel site at 2040246-2040259 bp of chromosome 34 of Ensembl chicken reference genome (GRCg7b version), the deleted nucleotide sequence is GACCCTGTGACACC (SEQ ID NO: 1), i.e. g.2040246_2040259delGACCCTGTGACACC;

[0102] There is a single nucleotide polymorphism at 2040608 bp of chromosome 34 of Ensembl chicken reference genome (GRCg7b version), which is G or A polymorphism, and there are GG and AA genotypes;

[0103] The InDel site is homozygous deletion type, and the genotype of the SNP site is AA, and the feather phenotype of the individual is curly feather;

[0104] The InDel site is homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is slice feather;

[0105] The InDel site is homozygous wild type or heterozygous type, and the feather phenotype of the individual is slice feather.

[0106] Example 5 A kit for identifying the feather trait of a chicken

[0107] I. Composition

[0108] The primer with the nucleotide sequence as shown in SEQ ID NO: 2-3, the primer with the nucleotide sequence as shown in SEQ ID NO: 4-5, 2x Es Taq MasterMix and ddH2O.

[0109] II. Method of use

[0110] The detection and result judgment are carried out according to example 4.

[0111] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited by the above examples, and any changes, modifications, substitutions, combinations and simplifications made without departing from the spirit and principles of the present application shall be equivalent replacement modes and shall be included in the protection scope of the present application.

Claims

1. Use of a composition for identifying feather traits of chickens, characterized in that: The composition is a detection reagent for InDel sites and SNP sites; The InDel site is located at 2040246-2040259 bp on chromosome 34 of the chicken genome, and the deleted nucleotide sequence is shown in SEQ ID NO: 1; The SNP site is located at position 2040608 on chromosome 34 of the chicken genome and is a G or A polymorphism, with GG and AA genotypes. The chicken genome is the GRCg7b version of the Ensembl database; The InDel site is a homozygous deletion type, and the feather phenotype of the individual whose genotype of the SNP site is AA is curly feathers; The InDel site is a homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is a piece feather; The feather phenotype of individuals whose InDel loci are homozygous wild type or heterozygous is a piece feather; The detection reagents for the InDel sites are primers with nucleotide sequences as shown in SEQ ID NOs: 2 to 3, and the detection reagents for the SNP sites are primers with nucleotide sequences as shown in SEQ ID NOs: 4 to 5; The chicken is the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a hybrid offspring of the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a bearded chicken as a parent.

2. Use of a kit comprising the detection reagent for the InDel site and the detection reagent for the SNP site according to claim 1 for identifying feather traits of chickens, characterized in that: The InDel site is located at 2040246-2040259 bp on chromosome 34 of the chicken genome, and the deleted nucleotide sequence is shown in SEQ ID NO: 1; The SNP site is located at position 2040608 on chromosome 34 of the chicken genome and is a G or A polymorphism, with GG and AA genotypes. The chicken genome is the GRCg7b version of the Ensembl database; The InDel site is a homozygous deletion type, and the feather phenotype of the individual whose genotype of the SNP site is AA is curly feathers; The InDel site is a homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is a piece feather; The feather phenotype of individuals whose InDel loci are homozygous wild type or heterozygous is a piece feather; The detection reagents for the InDel sites are primers with nucleotide sequences as shown in SEQ ID NOs: 2 to 3, and the detection reagents for the SNP sites are primers with nucleotide sequences as shown in SEQ ID NOs: 4 to 5; The chicken is the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a hybrid offspring of the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a bearded chicken as a parent.

3. Use of a kit comprising the detection reagent for the InDel site and the detection reagent for the SNP site according to claim 1 in preparing chicken breeding products, characterized in that: The breeding is molecular marker-assisted breeding of feather traits; The InDel site is located at 2040246-2040259 bp on chromosome 34 of the chicken genome, and the deleted nucleotide sequence is shown in SEQ ID NO: 1; The SNP site is located at position 2040608 on chromosome 34 of the chicken genome and is a G or A polymorphism, with GG and AA genotypes. The chicken genome is the GRCg7b version of the Ensembl database; The InDel site is a homozygous deletion type, and the feather phenotype of the individual whose genotype of the SNP site is AA is curly feathers; The InDel site is a homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is a piece feather; The feather phenotype of individuals whose InDel loci are homozygous wild type or heterozygous is a piece feather; The detection reagents for the InDel sites are primers with nucleotide sequences as shown in SEQ ID NOs: 2 to 3, and the detection reagents for the SNP sites are primers with nucleotide sequences as shown in SEQ ID NOs: 4 to 5; The chicken is the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a hybrid offspring of the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a bearded chicken as a parent.

4. The use according to claim 2 or 3, characterized in that The kit also contains PCR amplification reagents and ddH2O.

5. A method for identifying the feather characteristics of chickens, characterized in that: The following steps are involved: S1. Extract genomic DNA from the sample to be tested; S2. performing PCR amplification on the genomic DNA obtained in step S1 using the kit according to any one of claims 2 to 4, and then sequencing the PCR amplification product to obtain sequencing data; S3 analysis step S2 obtained sequencing data, according to the InDel site and SNP site, determine the feather traits of the chicken to be tested; The InDel site is located at 2040246-2040259 bp on chromosome 34 of the chicken genome, and the deleted nucleotide sequence is shown in SEQ ID NO: 1; The SNP site is located at position 2040608 on chromosome 34 of the chicken genome and is a G or A polymorphism, with GG and AA genotypes. The chicken genome is the GRCg7b version of the Ensembl database; The InDel site is a homozygous deletion type, and the feather phenotype of the individual whose genotype of the SNP site is AA is curly feathers; The InDel site is a homozygous deletion type, and the genotype of the SNP site is GG, and the feather phenotype of the individual is a piece feather; The feather phenotype of individuals whose InDel loci are homozygous wild type or heterozygous is a piece feather; The detection reagents for the InDel sites are primers with nucleotide sequences as shown in SEQ ID NOs: 2 to 3, and the detection reagents for the SNP sites are primers with nucleotide sequences as shown in SEQ ID NOs: 4 to 5; The chicken is the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a hybrid offspring of the western Guangdong curly-feathered chicken, Qibainong chicken, Wenchang chicken, Huaixiang chicken, Qingyuan hemp chicken, or a bearded chicken as a parent.

6. The method according to claim 5, characterized in that In step S2, the sequencing is Sanger sequencing.

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