CN117587133A - SNP locus combination for identifying goose variety and identification method thereof - Google Patents

Abstract

The invention discloses SNP locus combination for identifying a goose variety and an identification method thereof, and belongs to the technical field of biology. The invention screens out DA1P to DA15P 15 SNP loci, utilizes primer combinations for amplifying the SNP loci to determine genotypes of all loci, and judges the probability that the individual belongs to a goose according to the Bayesian theorem. Through experimental verification, the combined use of 15 SNP loci is proved to judge whether an individual to be detected belongs to a goose, and when the probability is greater than 0.95, the goose is judged to be a goose, and the accuracy rate can reach 100%. The method for identifying the goose variety is simple and convenient to operate and high in accuracy, and can effectively strike the degree of flooding of the counterfeit goose in the market.

Description

SNP locus combinations used to identify Ding'an goose breeds and their identification methods

Technical field

The present invention relates to the field of biotechnology, and in particular to a combination of SNP sites used to identify Ding'an goose varieties and an identification method thereof.

Background technique

Ding'an goose is a local breed in Hainan Province. It is a medium-sized goose species. It has the characteristics of tolerance to rough feeding, strong disease resistance, egg production in all seasons, and delicious meat. Its development and utilization prospects are great. Currently, there are many counterfeit Ding'an geese and hybrids of Ding'an goose and other breeds being sold on the market, which has had a certain impact on the purebred individual Ding'an goose market and seriously affected the Ding'an goose brand. Therefore, it is very necessary to provide a product and method that can effectively and accurately identify Ding'an goose species.

Currently, the method of identifying Ding'an geese on the market mainly relies on appearance observation. This kind of observation is highly subjective and has a high identification error rate. Especially when identifying the offspring of hybrids of Ding'an geese and other varieties of geese, it can easily lead to incorrect classification and identification. Moreover, local breed geese are relatively similar when they first hatch, and the identification error rate is also high.

Single nucleotide polymorphism (SNP) mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level. It has rich sites, wide distribution, high genetic stability, It is representative and has the characteristics of convenient and fast detection. SNP markers use the unique SNP sites in individual genomic DNA that have great variety differences to identify varieties through the genotypes of different site combinations, making the identification results more objective and accurate. In view of this, there is an urgent need in scientific research and practice to develop a product and method that can effectively and accurately identify Ding'an goose species, and apply them to the identification of Ding'an goose species.

Contents of the invention

The purpose of the present invention is to provide a combination of SNP sites for identifying Ding'an goose varieties and an identification method thereof, so as to solve the problems existing in the above-mentioned existing technologies, and jointly determine the potential candidates by detecting the genotypes of 15 SNP sites of the provided Ding'an goose. This method is easy to operate and has high accuracy to detect whether an individual belongs to Ding'an geese. It can effectively combat the proliferation of counterfeit Ding'an geese in the market.

In order to achieve the above objects, the present invention provides the following solutions:

The present invention provides a primer combination for identifying Ding'an goose varieties. The primer combination is a primer combination for amplifying the SNP site combination shown by DA1P-DA15P. The SNP site combination shown by DA1P-DA15P is:

DA1P is located at position 8129687 on chromosome NW_025927668.1, and its polymorphism is A or C;

DA2P is located at position 926886 on chromosome NW_025927673.1, and its polymorphism is C or G;

DA3P is located at position 936602 on chromosome NW_025927673.1, and its polymorphism is G or C;

DA4P is located at position 945751 on chromosome NW_025927673.1, and its polymorphism is C or A;

DA5P is located at position 946693 on chromosome NW_025927673.1, and its polymorphism is T or C;

DA6P is located at position 951840 on chromosome NW_025927673.1, and its polymorphism is A or G; DA7P is located at position 963963 on chromosome NW_025927673.1, and its polymorphism is G or C; DA8P is located at position 970250 on chromosome NW_025927673.1 , its polymorphism is G or A; DA9P is located at position 971259 on chromosome NW_025927673.1, and its polymorphism is T or A; DA10P is located at position 983590 on chromosome NW_025927673.1, and its polymorphism is T or A; DA11P is located at position 985387 on chromosome NW_025927673.1, and its polymorphism is C or T; DA12P is located at position 4763516 on chromosome NW_025927679.1, and its polymorphism is T or C; DA13P is located at position 1202481 on chromosome NW_025927866.1 , its polymorphism is A or C; DA14P is located at position 469618 on chromosome NW_025927879.1, and its polymorphism is T or C; DA15P is located at position 471621 on chromosome NW_025927879.1, and its polymorphism is C or T. Preferably, the amplification primer combination of the SNP site combination shown by DA1P-DA15P is: DA1F: 5’-TTAATTGCGTGCATCGAGCC-3’ (SEQ ID NO: 1);

DA1R: 5’-GTGATGGAAAAGGCCAGTGC-3’ (SEQ ID NO: 2); DA2F: 5’-CAGTTGGCTAAGCTGAGGGTG-3’ (SEQ ID NO: 3);

DA2R: 5’-GTTTGGATTCTGGGTCTCCCC-3’ (SEQ ID NO: 4);

DA3F: 5’-CTGACCTATCTGCACAGGGC-3’ (SEQ ID NO: 5);

DA3R: 5’-TCTGGGGCAGACAAGTGAAC-3’ (SEQ ID NO: 6);

DA4F: 5'-TACTTCGTTGTTGGTGACCCT-3' (SEQ ID NO: 7); DA4R: 5'-TTGGCCCAACCCATAAAACAC-3' (SEQ ID NO: 8); DA5F: 5'-GCATGAGCCATGCAGAGTTTC-3' (SEQ ID NO: 9); DA5R: 5'-AAAGTGCCAGAAGAGTATAGCCAA-3' (SEQ ID NO: 10);

DA6F: 5’-TGCCTGTGTCCATTGAAGT-3’ (SEQ ID NO: 11); DA6R: 5’-AGCAAGCGCAGAGCAAAATC-3’ (SEQ ID NO: 12);

DA7F: 5’-CTCTGCCCCATGAGCGTTAC-3’ (SEQ ID NO: 13); DA7R: 5’-TGGAGCTATGAGTGCCAAAGAA-3’ (SEQ ID NO: 14);

DA8F: 5’-GGTGTCAGGGGCTCTTTTCA-3’ (SEQ ID NO: 15);

DA8R: 5'-CATCAGTACCCAGCTCTGCC-3' (SEQ ID NO: 16); DA9F: 5'-AATTTGGACGGCACCTCACA-3' (SEQ ID NO: 17); DA9R: 5'-ATGGGCTCCCAGAAAGGGAA-3' (SEQ ID NO: 18); DA10F: 5'-AACCATAGCTCAGGTTTGCAC-3' (SEQ ID NO: 19);

DA10R: 5’-GAGCCATCAGAGCTAAAAGCAAG-3’ (SEQ ID NO: 20);

DA11F: 5’-TGCCTCTTGCAGTTCCATCAG-3’ (SEQ ID NO: 21);

DA11R: 5’-ACAAGGCAAAAAGTGGACATGC-3’ (SEQ ID NO: 22);

DA12F: 5’-CGGTATTATTGCAGTGCGGG-3’ (SEQ ID NO: 23);

DA12R: 5’-TCTTACACGGTGGCACGAAT-3’ (SEQ ID NO: 24);

DA13F: 5’-GAGTAAGAGCACCTGGCTGG-3’ (SEQ ID NO: 25);

DA13R: 5’-ATCTGGAGGCTCAGGAGAGG-3’ (SEQ ID NO: 26);

DA14F: 5’-TCTGCAGAGCTGTCACTTCA-3’ (SEQ ID NO: 27);

DA14R: 5’-CTCCCCGGCTTCATTGACTT-3’ (SEQ ID NO: 28);

DA15F: 5’-CAGGGGACTGTGCTATGGTG-3’ (SEQ ID NO: 29);

DA15R: 5'-GTTCTCACACATGCCAGGGA-3' (SEQ ID NO: 30).

The invention also provides a method for identifying Ding'an goose varieties, which includes the following steps:

Extract the total genomic DNA of the goose to be tested;

Using the total DNA as a template, use the primer combination to perform PCR amplification to obtain an amplification product;

Sequence the amplified product to determine the genotype of the sites shown from DA1P to DA15P;

According to the genotype measurement results, it is determined whether the goose to be tested belongs to Ding'an goose.

Preferably, the probability that the goose to be tested belongs to Ding'an goose is determined according to Bayes' theorem, and the calculation formula is as follows:

The probability that this individual belongs to the non-Ding'an goose is

Among them, p _i represents the frequency of the genotype corresponding to the i-th SNP in the SNP site and the Ding'an goose breed; q _i represents the average frequency of the genotype corresponding to the i-th SNP in the SNP site and other breeds.

Preferably, according to the calculation results, in order to ensure accuracy, 0.95 is used as the threshold. If the probability of belonging to a Ding'an goose is greater than or equal to 0.95, it is determined to be a Ding'an goose; if the probability of belonging to a non-Ding'an goose is greater than or equal to 0.95, it is determined to be a non-Ding'an goose; if less than 0.95, it can only be judged as one possibility. For example, the probability of belonging to Ding'an goose is 0.85, then it is considered that there is only 85% possibility of belonging to Ding'an goose.

Preferably, the genotypes and average genotype frequencies of the sites shown from DA1P to DA15P in Ding'an geese and other breeds are as follows:

DA1P site: When the genotype is AA, the genotype frequency of Ding'an goose is 0.72, and the genotype frequency of other breeds is 0.02; when the genotype is AC, the genotype frequency of Ding'an goose is 0.26, and the genotype frequency of other breeds is 0.26. The frequency is 0.21; when the genotype is CC, the genotype frequency of Ding'an goose is 0.02, and the genotype frequency of other breeds is 0.77;

DA2P site: When the genotype is CC, the genotype frequency of Ding'an goose is 0.49, and the genotype frequency of other breeds is 0.00; when the genotype is CG, the genotype frequency of Ding'an goose is 0.42, and the genotype frequency of other breeds is 0.42. The frequency is 0.06; when the genotype is GG, the genotype frequency of Ding'an goose is 0.09, and the genotype frequency of other breeds is 0.94;

DA3P site: When the genotype is GG, the genotype frequency of Ding'an goose is 0.56, and the genotype frequency of other breeds is 0.00; when the genotype is GC, the genotype frequency of Ding'an goose is 0.38, and the genotype frequency of other breeds is 0.38. The frequency is 0.07; when the genotype is CC, the genotype frequency of Ding'an goose is 0.06, and the genotype frequency of other breeds is 0.93;

DA4P site: When the genotype is CC, the genotype frequency of Ding'an goose is 0.49, and the genotype frequency of other breeds is 0.00; when the genotype is CA, the genotype frequency of Ding'an goose is 0.42, and the genotype frequency of other breeds is 0.42. The frequency is 0.08; when the genotype is AA, the genotype frequency of Ding'an goose is 0.09, and the genotype frequency of other breeds is 0.92;

DA5P site: When the genotype is TT, the genotype frequency of Ding'an goose is 0.56, and the genotype frequency of other breeds is 0.00; when the genotype is TC, the genotype frequency of Ding'an goose is 0.38, and the genotype frequency of other breeds is 0.38. The frequency is 0.06; when the genotype is CC, the genotype frequency of Ding'an goose is 0.06, and the genotype frequency of other breeds is 0.94;

DA6P site: When the genotype is AA, the genotype frequency of Ding'an goose is 0.45, and the genotype frequency of other breeds is 0.00; when the genotype is AG, the genotype frequency of Ding'an goose is 0.44, and the genotype frequency of other breeds is 0.44. The frequency is 0.07; when the genotype is GG, the genotype frequency of Ding'an goose is 0.11, and the genotype frequency of other breeds is 0.93;

DA7P site: When the genotype is GG, the genotype frequency of Ding'an goose is 0.52, and the genotype frequency of other breeds is 0.00; when the genotype is GC, the genotype frequency of Ding'an goose is 0.40, and the genotype frequency of other breeds is 0.40. The frequency is 0.08; when the genotype is CC, the genotype frequency of Ding'an goose is 0.08, and the genotype frequency of other breeds is 0.92;

DA8P site: When the genotype is GG, the genotype frequency of Ding'an goose is 0.56, and the genotype frequency of other breeds is 0.00; when the genotype is GA, the genotype frequency of Ding'an goose is 0.38, and the genotype frequency of other breeds is 0.38. The frequency is 0.10; when the genotype is AA, the genotype frequency of Ding'an goose is 0.06, and the genotype frequency of other breeds is 0.90;

DA9P site: When the genotype is TT, the genotype frequency of Ding'an goose is 0.52, and the genotype frequency of other breeds is 0.00; when the genotype is TA, the genotype frequency of Ding'an goose is 0.40, and the genotype frequency of other breeds is 0.40. The frequency is 0.09; when the genotype is AA, the genotype frequency of Ding'an goose is 0.08, and the genotype frequency of other breeds is 0.91;

DA10P site: When the genotype is TT, the genotype frequency of Ding'an goose is 0.56, and the genotype frequency of other breeds is 0.00; when the genotype is TA, the genotype frequency of Ding'an goose is 0.38, and the genotype frequency of other breeds is 0.38. The frequency is 0.07; when the genotype is AA, the genotype frequency of Ding'an goose is 0.06, and the genotype frequency of other breeds is 0.93;

DA11P locus: When the genotype is CC, the genotype frequency of Ding'an goose is 0.52, and the genotype frequency of other breeds is 0.00; when the genotype is CT, the genotype frequency of Ding'an goose is 0.40, and the genotype frequency of other breeds is 0.40. The frequency is 0.07; when the genotype is TT, the genotype frequency of Ding'an goose is 0.08, and the genotype frequency of other breeds is 0.93;

DA12P site: When the genotype is TT, the genotype frequency of Ding'an goose is 0.81, and the genotype frequency of other breeds is 0.01; when the genotype is TC, the genotype frequency of Ding'an goose is 0.18, and the genotype frequency of other breeds The frequency is 0.20; when the genotype is CC, the genotype frequency of Ding'an goose is 0.01, and the genotype frequency of other breeds is 0.79;

DA13P site: When the genotype is AA, the genotype frequency of Ding'an goose is 0.60, and the genotype frequency of other breeds is 0.00; when the genotype is AC, the genotype frequency of Ding'an goose is 0.35, and the genotype frequency of other breeds is 0.35. The frequency is 0.15; when the genotype is CC, the genotype frequency of Ding'an goose is 0.05, and the genotype frequency of other breeds is 0.85;

DA14P site: When the genotype is TT, the genotype frequency of Ding'an goose is 0.72, and the genotype frequency of other breeds is 0.02; when the genotype is TC, the genotype frequency of Ding'an goose is 0.26, and the genotype frequency of other breeds is 0.26. The frequency is 0.25; when the genotype is CC, the genotype frequency of Ding'an goose is 0.02, and the genotype frequency of other breeds is 0.73;

DA15P locus: When the genotype is CC, the genotype frequency of Ding'an goose is 0.77, and the genotype frequency of other breeds is 0.02; when the genotype is CT, the genotype frequency of Ding'an goose is 0.22, and the genotype frequency of other breeds is 0.22. The frequency is 0.25; when the genotype is TT, the genotype frequency of Ding'an goose is 0.01, and the genotype frequency of other breeds is 0.73.

Preferably, the other species include Changle goose, Huoyan goose, Daozhou gray goose, steel goose, Lingxian white goose, lion head goose, black mane goose, Yongkang gray goose and seed goose.

The invention also provides a kit for identifying Ding'an goose species, including the primer combination.

The present invention also provides the application of the primer combination or the kit in identifying Ding'an goose varieties.

The invention discloses the following technical effects:

The invention discloses 15 SNP sites. According to Bayes' theorem, the probability that the individual belongs to the Ding'an goose can be determined by combining the 15 SNP sites with the genotype. The present invention combines 15 SNP sites to determine the probability of Ding'an goose up to 100%. This method is simple to operate and highly accurate, and can effectively combat the proliferation of counterfeit Ding'an geese in the market.

Detailed ways

Various exemplary embodiments of the invention will now be described in detail. This detailed description should not be construed as limitations of the invention, but rather as a more detailed description of certain aspects, features and embodiments of the invention.

It should be understood that the terms used in the present invention are only used to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges in the present invention, it should be understood that every intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or value intermediate within a stated range, and any other stated value or value intermediate within a stated range, is also included within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded from the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only the preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials in connection with which the documents relate. In the event of conflict with any incorporated document, the contents of this specification shall prevail.

It will be apparent to those skilled in the art that various modifications and changes can be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to the skilled person from the description of the invention. The specification and examples of the present invention are exemplary only.

The words "includes", "includes", "has", "contains", etc. used in this article are all open terms, which mean including but not limited to.

The present invention provides a primer combination for identifying Ding'an goose varieties. The primer combination is a primer combination for amplifying the following SNP site combinations shown from DA1P to DA15P (see Table 1):

Table 1

serial number chromosome site major allele minor allele DA1P NW_025927668.1 8129687 A C DA2P NW_025927673.1 926886 C G DA3P NW_025927673.1 936602 G C DA4P NW_025927673.1 945751 C A DA5P NW_025927673.1 946693 T C DA6P NW_025927673.1 951840 A G DA7P NW_025927673.1 963963 G C DA8P NW_025927673.1 970250 G A DA9P NW_025927673.1 971259 T A DA10P NW_025927673.1 983590 T A DA11P NW_025927673.1 985387 C T DA12P NW_025927679.1 4763516 T C DA13P NW_025927866.1 1202481 A C DA14P NW_025927879.1 469618 T C DA15P NW_025927879.1 471621 C T

The screening method for SNP site combinations shown from DA1P to DA15P is: first determine Changle goose, Huoyan goose, Daozhou gray goose, steel goose, Lingxian white goose, lion head goose, black mane goose, Yongkang gray goose, seed goose Then, compare the mean value with the different allele frequencies of each site in Ding'an geese to screen out SNP sites with large differences in allele frequencies. , the allele frequency of a certain site in Ding'an geese is preferably 0 or 1, and the allele frequency in other populations is close to 1 or close to 0.

The primer combinations used to amplify the DA1P to DA15P locus are as follows, see Table 2:

Table 2

The invention provides a method for identifying Ding'an goose varieties, which includes the following steps:

Extract the total genomic DNA of the goose to be tested;

Using the total DNA as a template, perform PCR amplification using the above primer combination to obtain an amplification product;

Sequence the amplification product to determine the genotype;

According to the genotype measurement results, it is determined whether the goose to be tested belongs to Ding'an goose.

In the identification method of the present invention, genomic DNA is preferably obtained by collecting blood from the wing vein of the goose to be tested. The present invention has no special restrictions on the extraction method of genomic DNA. Conventional extraction methods in the field can be used. In the embodiments of the present invention, phenol is preferred. - Extraction of genomic DNA using chloroform method.

In the present invention, the combination of sites in DA1P to DA15P is used to identify Ding'an goose varieties, and the corresponding upstream and downstream primers are used for PCR amplification. The present invention has no special limitations on the PCR amplification method, and conventional PCR amplification methods in this field can be used.

In the present invention, by sequencing the PCR amplification product of the goose to be tested, determining the genotype, and comparing it with the genotype corresponding to the selected SNP combination, the probability that the goose to be tested belongs to the Ding'an goose is determined according to Bayes' theorem. The calculation formula is as follows :

The probability that the goose to be tested is a non-Ding'an goose is

Among them, p _i represents the frequency of genotypes corresponding to the Ding'an goose variety in the i-th SNP in the SNP combination; q _i represents the average frequency of genotypes corresponding to other varieties in the i-th SNP in the SNP combination. Considering the genotyping error rate and the extreme situation that when the genotype frequency is 0, the product of frequencies is 0, genotype frequencies less than 0.05 are uniformly adjusted to 0.05, and the maximum genotype frequency is adjusted to ensure that the frequency of the three genotypes is equal to 0. The sum is 1.

According to the calculation results, in order to ensure accuracy, 0.95 is used as the threshold. If the probability of belonging to a Dingan goose is greater than or equal to 0.95, it is determined to be a Dingan goose; if the probability of belonging to a non-Ding'an goose is greater than or equal to 0.95, it is determined to be a non-Ding'an goose; if it is less than 0.95, only It can be judged as a possibility. For example, the probability of belonging to Ding'an goose is 0.85, then it is considered that there is only 85% possibility of belonging to Ding'an goose.

The genotypes and genotype frequencies of the above SNP sites in Ding'an geese and other breeds are as follows, see Table 3:

table 3

For example, if 15 SNP sites from DA1P to DA15P are selected, and the combined genotype is AA CC CC CA TC AAGG GG TT TT CC TC AA TC CT, then the probability that the individual belongs to the Ding'an goose is:

The probability that this individual belongs to the non-Ding'an goose is

Among them, the genotype frequency of other varieties refers to the selection of domestic representative varieties such as Changle goose, Huoyan goose, Daozhou gray goose, steel goose, Lingxian white goose, lion head goose, black mane goose, Yongkang gray goose, and seed goose. , the average genotype frequency of each locus calculated from different varieties. The genotype frequency of each locus was calculated separately for Ding'an geese.

In practical application, the present invention can also be prepared into a kit for identifying Ding'an goose species. The kit includes the above primer combination and other conventional reagents for detection.

The present invention also provides the application of the above-mentioned primer combination or the above-mentioned kit in identifying Ding'an goose varieties.

In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below with reference to specific embodiments, but they should not be understood as limiting the scope of the present invention.

Example 1

Randomly select 10 Ding'an geese, and randomly select 10 geese from other varieties (Changle geese, Huoyan geese, Daozhou gray geese, steel geese, Lingxian white geese, lion-head geese, black-maned geese, Yongkang gray geese, and seed geese) Only, blood was collected from the wing vein, genomic DNA was extracted using the phenol-chloroform method, and PCR amplification was performed based on the primers of 15 SNP sites in Table 2.

The PCR system (20 μL) is: 2 μL of DNA template, 1.5 μL of forward and reverse primers (10 ng/μL), 10 μL of 2× PCR reagent in Tiangen PCR kit, and the rest is made up with ultrapure water.

The PCR program was: first denaturation at 94°C for 1 min, then 94°C for 30 s, annealing at 55°C for 30 s, and extension at 72°C for 30 s, a total of 35 cycles, and finally extension at 72°C for 5 min, and then stored at 4°C. The PCR instrument was the American Bio-Rad T100 gradient PCR instrument.

The amplified products were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequence polymorphism detection and genotype determination. According to the sequencing results, 15 SNP sites combined with the genotype were used to determine the probability of belonging to the Dingan goose. Taking 0.95 as the threshold, the probability of belonging to the Dingan goose was greater than 0.95, and it was determined to be a Dingan goose. As a result, all 10 Ding'an geese were judged correctly; they were non-Ding'an geese. The probability of Ding'an geese is greater than 0.95, and it is judged as non-Ding'an geese. As a result, all 10 non-Ding'an geese were judged correctly. The results are shown in Table 4 below.

Table 4

The above-described embodiments only describe the preferred modes of the present invention and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, those of ordinary skill in the art can make various modifications to the technical solutions of the present invention. All deformations and improvements shall fall within the protection scope determined by the claims of the present invention.

Claims (9)

1. A primer combination for identifying a goose variety, which is characterized in that the primer combination is a primer combination for amplifying SNP locus combination shown by DA1P-DA15P, and the SNP locus combination shown by DA1P-DA15P is as follows:

DA1P is located at 8129687 on NW_025927668.1 chromosome and has polymorphism A or C;

DA2P is located at 926886 on NW_025927673.1 chromosome and has polymorphism C or G;

DA3P is located at 936602 on NW_025927673.1 chromosome and has G or C polymorphism;

DA4P is located at 945751 on NW_025927673.1 chromosome and has polymorphism C or A;

DA5P is located at 946693 on NW_025927673.1 chromosome and has a polymorphism of T or C;

DA6P is located at 951840 on NW_025927673.1 chromosome and has polymorphism A or G;

DA7P is located at 963963 on NW_025927673.1 chromosome and has G or C polymorphism;

DA8P is located at 970250 on NW_025927673.1 chromosome and has a polymorphism of G or A;

DA9P is located at 971259 on NW_025927673.1 chromosome and has a polymorphism of T or A;

DA10P is located at 983590 on NW_025927673.1 chromosome and has polymorphism T or A;

DA11P is located at 985387 on NW_025927673.1 chromosome and has polymorphism C or T;

DA12P is located at 4763516 on NW_025927679.1 chromosome and has a polymorphism of T or C;

DA13P is located at 1202481 on NW_025927866.1 chromosome, and its polymorphism is A or C;

DA14P is located at 469618 on NW_025927879.1 chromosome and has a polymorphism of T or C;

DA15P is located at position 471621 on the NW_025927879.1 chromosome and has a polymorphism of C or T.

2. The primer combination according to claim 1, wherein the amplification primer combination of the SNP site combination shown in DA1P-DA15P is:

DA1F：5’-TTAATTGCGTGCATCGAGCC-3’；

DA1R：5’-GTGATGGAAAAGGCCAGTGC-3’；

DA2F：5’-CAGTTGGCTAAGCTGAGGGTG-3’；

DA2R：5’-GTTTGGATTCTGGGTCTCCCC-3’；

DA3F：5’-CTGACCTATCTGCACAGGGC-3’；

DA3R：5’-TCTGGGGCAGACAAGTGAAC-3’；

DA4F：5’-TACTTCGTTGTTGGTGACCCT-3’；

DA4R：5’-TTGGCCCAACCCATAAAACAC-3’；

DA5F：5’-GCATGAGCCATGCAGAGTTTC-3’；

DA5R：5’-AAAGTGCCAGAAGAGTATAGCCAA-3’；

DA6F：5’-TGCCTGTGTCCCATTGAAGT-3’；

DA6R：5’-AGCAAGCGCAGAGCAAAATC-3’；

DA7F：5’-CTCTGCCCCATGAGCGTTAC-3’；

DA7R：5’-TGGAGCTATGAGTGCCAAAGAA-3’；

DA8F：5’-GGTGTCAGGGGCTCTTTTCA-3’；

DA8R：5’-CATCAGTACCCAGCTCTGCC-3’；

DA9F：5’-AATTTGGACGGCACCTCACA-3’；

DA9R：5’-ATGGGCTCCCAGAAAGGGAA-3’；

DA10F：5’-AACCATAGCTCAGGTTTGCAC-3’；

DA10R：5’-GAGCCATCAGAGCTAAAAGCAAG-3’；

DA11F：5’-TGCCTCTTGCAGTTCCATCAG-3’；

DA11R：5’-ACAAGGCAAAAAGTGGACATGC-3’；

DA12F：5’-CGGTATTATTGCAGTGCGGG-3’；

DA12R：5’-TCTTACACGGTGGCACGAAT-3’；

DA13F：5’-GAGTAAGAGCACCTGGCTGG-3’；

DA13R：5’-ATCTGGAGGCTCAGGAGAGG-3’；

DA14F：5’-TCTGCAGAGCTGTCACTTCA-3’；

DA14R：5’-CTCCCCGGCTTCATTGACTT-3’；

DA15F：5’-CAGGGGACTGTGCTATGGTG-3’；

DA15R：5’-GTTCTCACACATGCCAGGGA-3’。

3. a method for identifying a goose variety, comprising the steps of:

extracting total DNA of a goose genome to be detected;

performing PCR amplification by using the primer combination according to claim 1 or 2 with the total DNA as a template to obtain an amplification product;

sequencing the amplification product, and judging genotypes of sites shown by DA1P to DA 15P;

and judging whether the goose to be detected belongs to a goose in the goose mounting state according to the genotype determination result.

4. A method according to claim 3, characterized in that the probability that the goose to be tested belongs to a goose with a mounted goose is determined according to the bayesian theorem, and the calculation formula is as follows:

the probability that the goose to be detected belongs to the non-mounted goose is calculated as follows:

wherein p is _i Representing the frequency of the genotype corresponding to the goose variety in the ith SNP in the SNP locus combination; q _i The average frequency of genotypes corresponding to other varieties in the ith SNP in the SNP locus combination is shown.

5. The method of claim 4, wherein, according to the calculation result, in order to ensure accuracy, 0.95 is taken as a threshold value, and the probability of belonging to the goose is greater than or equal to 0.95, and the goose is determined to be the goose; and judging that the goose is not mounted if the probability of the goose being not mounted is more than or equal to 0.95.

6. The method of claim 3, wherein the genotypes at the sites shown by DA1P to DA15P in geese and other varieties, and the average frequencies of genotypes are as follows:

DA1P site: when the genotype is AA, the genotype frequency of the goose is 0.72, and the genotype frequency of other varieties is 0.02; when the genotype is AC, the genotype frequency of the goose is 0.26, and the genotype frequency of other varieties is 0.21; when the genotype is CC, the genotype frequency of the goose is 0.02, and the genotype frequency of other varieties is 0.77;

DA2P site: when the genotype is CC, the genotype frequency of the goose is 0.49, and the genotype frequency of other varieties is 0.00; when the genotype is CG, the genotype frequency of the goose is 0.42, and the genotype frequency of other varieties is 0.06; when the genotype is GG, the genotype frequency of the goose is 0.09, and the genotype frequency of other varieties is 0.94;

DA3P site: when the genotype is GG, the genotype frequency of the goose is 0.56, and the genotype frequency of other varieties is 0.00; when the genotype is GC, the genotype frequency of the goose is 0.38, and the genotype frequency of other varieties is 0.07; when the genotype is CC, the genotype frequency of the goose is 0.06, and the genotype frequency of other varieties is 0.93;

DA4P site: when the genotype is CC, the genotype frequency of the goose is 0.49, and the genotype frequency of other varieties is 0.00; when the genotype is CA, the genotype frequency of the goose is 0.42, and the genotype frequency of other varieties is 0.08; when the genotype is AA, the genotype frequency of the goose is 0.09, and the genotype frequency of other varieties is 0.92;

DA5P site: when the genotype is TT, the genotype frequency of the goose is 0.56, and the genotype frequency of other varieties is 0.00; when the genotype is TC, the genotype frequency of the goose is 0.38, and the genotype frequency of other varieties is 0.06; when the genotype is CC, the genotype frequency of the goose is 0.06, and the genotype frequency of other varieties is 0.94;

DA6P site: when the genotype is AA, the genotype frequency of the goose is 0.45, and the genotype frequency of other varieties is 0.00; when the genotype is AG, the genotype frequency of the goose is 0.44, and the genotype frequency of other varieties is 0.07; when the genotype is GG, the genotype frequency of the goose is 0.11, and the genotype frequency of other varieties is 0.93;

DA7P site: when the genotype is GG, the genotype frequency of the goose is 0.52, and the genotype frequency of other varieties is 0.00; when the genotype is GC, the genotype frequency of the goose is 0.40, and the genotype frequency of other varieties is 0.08; when the genotype is CC, the genotype frequency of the goose is 0.08, and the genotype frequency of other varieties is 0.92;

DA8P site: when the genotype is GG, the genotype frequency of the goose is 0.56, and the genotype frequency of other varieties is 0.00; when the genotype is GA, the genotype frequency of the goose is 0.38, and the genotype frequency of other varieties is 0.10; when the genotype is AA, the genotype frequency of the goose is 0.06, and the genotype frequency of other varieties is 0.90;

DA9P site: when the genotype is TT, the genotype frequency of the goose is 0.52, and the genotype frequency of other varieties is 0.00; when the genotype is TA, the genotype frequency of the goose is 0.40, and the genotype frequency of other varieties is 0.09; when the genotype is AA, the genotype frequency of the goose is 0.08, and the genotype frequency of other varieties is 0.91;

DA10P site: when the genotype is TT, the genotype frequency of the goose is 0.56, and the genotype frequency of other varieties is 0.00; when the genotype is TA, the genotype frequency of the goose is 0.38, and the genotype frequency of other varieties is 0.07; when the genotype is AA, the genotype frequency of the goose is 0.06, and the genotype frequency of other varieties is 0.93;

DA11P site: when the genotype is CC, the genotype frequency of the goose is 0.52, and the genotype frequency of other varieties is 0.00; when the genotype is CT, the genotype frequency of the goose is 0.40, and the genotype frequency of other varieties is 0.07; when the genotype is TT, the genotype frequency of the goose is 0.08, and the genotype frequency of other varieties is 0.93;

DA12P site: when the genotype is TT, the genotype frequency of the goose is 0.81, and the genotype frequency of other varieties is 0.01; when the genotype is TC, the genotype frequency of the goose is 0.18, and the genotype frequency of other varieties is 0.20; when the genotype is CC, the genotype frequency of the goose is 0.01, and the genotype frequency of other varieties is 0.79;

DA13P site: when the genotype is AA, the genotype frequency of the goose is 0.60, and the genotype frequency of other varieties is 0.00; when the genotype is AC, the genotype frequency of the goose is 0.35, and the genotype frequency of other varieties is 0.15; when the genotype is CC, the genotype frequency of the goose is 0.05, and the genotype frequency of other varieties is 0.85;

DA14P site: when the genotype is TT, the genotype frequency of the goose is 0.72, and the genotype frequency of other varieties is 0.02; when the genotype is TC, the genotype frequency of the goose is 0.26, and the genotype frequency of other varieties is 0.25; when the genotype is CC, the genotype frequency of the goose is 0.02, and the genotype frequency of other varieties is 0.73;

DA15P site: when the genotype is CC, the genotype frequency of the goose is 0.77, and the genotype frequency of other varieties is 0.02; when the genotype is CT, the genotype frequency of the goose is 0.22, and the genotype frequency of other varieties is 0.25; when the genotype is TT, the genotype frequency of the goose is 0.01, and the genotype frequency of other varieties is 0.73.

7. The method of claim 4 or 6, wherein the other varieties comprise geese, geese in the open eye, geese in the state of the road, geese in the steel, geese in the county, geese in the lion head, geese in the black, geese in the Yongkang, and geese in the seed.

8. A kit for identifying a goose variety, comprising the primer combination of claim 1 or 2.

9. Use of a primer combination according to any one of claims 1-2 or a kit according to claim 8 for identifying a goose variety.