Genetic markers for individual identification of silver pheasant and its application

Abstract

The present invention relates to a genetic marker for individual identification of white pheasants and an application thereof, and relates to the field of biotechnology. The genetic marker comprises at least 20 of the SNP sites 1-86. The genetic marker is used to perform individual identification of white pheasants, and the individual identification result is accurate and reliable. Based on the individual identification result, kinship, population genetic diversity, and population genetic structure analysis can also be performed, meeting the protection research needs of the white pheasant species.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, in particular to genetic markers for individual identification of silver pheasants and applications thereof. Background Art

[0002] The Silver Pheasant (Lophura nycthemera), also known as the Silver Pheasant, Silver Pheasant, Yue Bird, and White Pheasant, is a member of the Pheasant genus of the Phasianidae family. It is widely distributed in southern China and Southeast Asia and is now a Class II key protected animal in my country. The Silver Pheasant is a forest-dwelling bird, mainly living under forests at an altitude of 0-1000 meters. However, its habitat has been severely damaged in recent years. The Silver Pheasant is a ground-dwelling bird with poor flying ability, weak ability to resist natural enemies, and a particularly high dependence on the living environment. Therefore, the loss and fragmentation of the habitat have become the main threats to the Silver Pheasant. At the same time, because the Silver Pheasant is sexually dimorphic and has beautiful feathers, it has long been threatened by indiscriminate hunting, resulting in a gradual decrease in its number. Therefore, it is urgent to carry out conservation biology research on its wild population to maintain the viability of its population.

[0003] Although the silver pheasant species is widely distributed in China, it is now rare or has disappeared in some places where it originally existed. Expanding the population through artificial breeding and releasing it into the natural environment where it was originally distributed to restore the population is one of the most effective ways to protect the silver pheasant, and it is also the ultimate goal and highest level of endangered species protection. However, for the wild population and reintroduced individuals of the silver pheasant, they are currently generally only captured by infrared cameras, and individual identification is impossible, making it difficult to assess the population size and genetic diversity. Therefore, it is necessary to develop a convenient and accurate molecular method to form a molecular tag for individual silver pheasants for identification, and then track the individual activities and diffusion range of the white pheasants, while also analyzing the genetic diversity of its population, and being able to evaluate the survival status and development trend of the white pheasant population. Summary of the invention

[0004] In view of the above problems, the present invention provides a genetic marker for individual identification of silver pheasants. The genetic marker is used to perform individual identification of silver pheasants. The individual identification results are accurate and reliable. Based on the individual identification results, kinship, population genetic diversity, and group genetic structure analysis can also be performed to meet the protection research needs of the silver pheasant species.

[0005] In order to achieve the above object, the present invention provides a genetic marker for individual identification of white pheasant, which includes at least 20 of the following SNP sites:

[0006] The SNP site 1 is located in the white pheasant genome JADANK010000113.1:5324820, and its base is C or T;

[0007] The SNP site 2 is located in the white pheasant genome JADANK010000063.1:10039215, and its base is C or G;

[0008] The SNP site 3 is located in the white pheasant genome JADANK010000024.1:15904660, and its base is C or T;

[0009] The SNP site 4 is located in the white pheasant genome JADANK010001474.1:10085797, and its base is G or T;

[0010] The SNP site 5 is located in the white pheasant genome JADANK010001487.1:10047134, and its base is A or G;

[0011] The SNP site 6 is located in the white pheasant genome JADANK010001500.1:10081764, and its base is G or A;

[0012] The SNP site 7 is located in the white pheasant genome JADANK010001509.1:301199, and its base is C or T;

[0013] The SNP site 8 is located in the white pheasant genome JADANK010000146.1:13374829, and its base is A or G;

[0014] The SNP site 9 is located in the white pheasant genome JADANK010001524.1:10055431, and its base is G or A;

[0015] The SNP site 10 is located in the white pheasant genome JADANK010001544.1:10002213, and its base is T or C;

[0016] The SNP site 11 is located in the white pheasant genome JADANK010001485.1:5025375, and its base is A or G;

[0017] The SNP site 12 is located in the white pheasant genome JADANK010001510.1:10226541, and its base is G or C;

[0018] The SNP site 13 is located in the white pheasant genome JADANK010000033.1:5079977, and its base is T or C;

[0019] The SNP site 14 is located in the white pheasant genome JADANK010001522.1:5612065, and its base is G or A;

[0020] The SNP site 15 is located in the white pheasant genome JADANK010001502.1:10380701, and its base is A or G;

[0021] The SNP site 16 is located in the white pheasant genome JADANK010001472.1:10421851, and its base is A or G;

[0022] The SNP site 17 is located in the white pheasant genome JADANK010000316.1:17079, and its base is C or T;

[0023] The SNP site 18 is located in the white pheasant genome JADANK010001547.1:480922, and its base is A or G;

[0024] The SNP site 19 is located in the white pheasant genome JADANK010000006.1:10748586, and its base is T or C;

[0025] The SNP site 20 is located in the white pheasant genome JADANK010001468.1:10617303, and its base is G or A;

[0026] The SNP site 21 is located in the white pheasant genome JADANK010001469.1:10711749, and its base is C or T;

[0027] The SNP site 22 is located in the white pheasant genome JADANK010001482.1:5142234, and its base is A or G;

[0028] The SNP site 23 is located in the white pheasant genome JADANK010001511.1:5123818, and its base is C or T;

[0029] The SNP site 24 is located in the white pheasant genome JADANK010001470.1:5597501, and its base is A or G;

[0030] The SNP site 25 is located in the white pheasant genome JADANK010001521.1:5015305, and its base is C or T;

[0031] The SNP site 26 is located in the white pheasant genome JADANK010001492.1:261381, and its base is C or T;

[0032] The SNP site 27 is located in the white pheasant genome JADANK010001490.1:2096, and its base is G or T;

[0033] The SNP site 28 is located in the white pheasant genome JADANK010001471.1: 5505053, and its base is C or T; the SNP site 29 is located in the white pheasant genome JADANK010000102.1: 6442137, and its base is A or G; the SNP site 30 is located in the white pheasant genome JADANK010000003.1: 823633, and its base is C or T; the SNP site 31 is located in the white pheasant genome JADANK010000004.1: 6106541, and its base is G or A; the SNP site 32 is located in the white pheasant genome JADANK010001465.1: 6256108, and its base is A or T; the SNP site 33 is located in the white pheasant genome JADANK010000032.1:15474, and its base is G or A; the SNP site 34 is located in the white pheasant genome JADANK010001506.1:5837633, and its base is T or C; the SNP site 35 is located in the white pheasant genome JADANK010001466.1:6605868, and its base is C or T; the SNP site 36 is located in the white pheasant genome JADANK010001467.1:7069787, and its base is C or T; the SNP site 37 is located in the white pheasant genome JADANK010001489.1:637286, and its base is is T or C; the SNP site 38 is located in the white pheasant genome JADANK010001550.1:64201, and its base is G or A; the SNP site 39 is located in the white pheasant genome JADANK010000001.1:111955, and its base is A or G; the SNP site 40 is located in the white pheasant genome JADANK010000002.1:316812, and its base is G or C; the SNP site 41 is located in the white pheasant genome JADANK010000005.1:73154, and its base is T or C; the SNP site 42 is located in the white pheasant genome JADANK010000008.1:123053, and its base is T or C; the SNP site 43 is located in the white pheasant genome JADANK010000019.1:138662, and its base is T or C; the SNP site 44 is located in the white pheasant genome JADANK010000021.1:473931, and its base is T or C; the SNP site 45 is located in the white pheasant genome JADANK010000022.1:151474, and its base is T or G; the SNP site 46 is located in the white pheasant genome JADANK010000023.1:238776, and its base is C or T; the SNP site 47 is located in the white pheasant genome JADANK010000029.1:1805, and its base is T or C;

[0034] The SNP site 48 is located in the white pheasant genome JADANK010000030.1:22918, and its base is C or T; the SNP site 49 is located in the white pheasant genome JADANK010000031.1:38390, and its base is T or C; the SNP site 50 is located in the white pheasant genome JADANK010000034.1:359, and its base is C or T;

[0035] The SNP site 51 is located in the white pheasant genome JADANK010000046.1:4067, and its base is G or A;

[0036] The SNP site 52 is located in the white pheasant genome JADANK010000048.1:74254, and its base is T or C; the SNP site 53 is located in the white pheasant genome JADANK010000050.1:196194, and its base is T or C; the SNP site 54 is located in the white pheasant genome JADANK010000061.1:1531940, and its base is A or G; the SNP site 55 is located in the white pheasant genome JADANK010000062.1:212481, and its base is C or T; the SNP site 56 is located in the white pheasant genome JADANK010000066.1:134569, and its base is The base is G or A; the SNP site 57 is located in the white pheasant genome JADANK010000067.1:133257, and its base is T or C; the SNP site 58 is located in the white pheasant genome JADANK010000069.1:155575, and its base is T or C; the SNP site 59 is located in the white pheasant genome JADANK010000070.1:65857, and its base is C or T; the SNP site 60 is located in the white pheasant genome JADANK010000071.1:281355, and its base is A or G; the SNP site 61 is located in the white pheasant genome JADANK010000073.1:11001 7, its base is G or A; the SNP site 62 is located in the white pheasant genome JADANK010000074.1:76097, its base is A or G; the SNP site 63 is located in the white pheasant genome JADANK010000086.1:167039, its base is T or C; the SNP site 64 is located in the white pheasant genome JADANK010000087.1:140185, its base is C or T; the SNP site 65 is located in the white pheasant genome JADANK010000091.1:582446, its base is C or T; the SNP site 66 is located in the white pheasant genome JADANK010000092.1: 12529, its base is T or C; the SNP site 67 is located in the white pheasant genome JADANK010000100.1:20025, its base is A or G; the SNP site 68 is located in the white pheasant genome JADANK010000105.1:145727, its base is G or A; the SNP site 69 is located in the white pheasant genome JADANK010000106.1:32240, its base is T or C; the SNP site 70 is located in the white pheasant genome JADANK010000116.1:261398, its base is C or T; the SNP site 71 is located in the white pheasant genome JADANK010000144.1:54717, its base is C or T; the SNP site 72 is located in the white pheasant genome JADANK010001473.1:25594, its base is A or G; the SNP site 73 is located in the white pheasant genome JADANK010001486.1:111080, its base is A or G; the SNP site 74 is located in the white pheasant genome JADANK010001488.1:82694, its base is A or G; the SNP site 75 is located in the white pheasant genome JADANK010001491.1:73650, its base is T or C; the SNP site 76 is located in the white pheasant genome JADANK010001501.1:101470, its The base is A or G; the SNP site 77 is located in the white pheasant genome JADANK010001504.1:66945, and its base is C or T; the SNP site 78 is located in the white pheasant genome JADANK010001507.1:13120, and its base is C or T; the SNP site 79 is located in the white pheasant genome JADANK010001526.1:275885, and its base is C or A; the SNP site 80 is located in the white pheasant genome JADANK010001531.1:233119, and its base is T or C; the SNP site 81 is located in the white pheasant genome JADANK010001533.1:1767861, and its base is T or C;.

[0037] The SNP site 82 is located in the white pheasant genome JADANK010001534.1:134674, and its base is A or G;

[0038] The SNP site 83 is located in the white pheasant genome JADANK010001545.1:87524, and its base is G or A;

[0039] The SNP site 84 is located in the white pheasant genome JADANK010001546.1:84383, and its base is A or C;

[0040] The SNP site 85 is located in the white pheasant genome JADANK010001548.1:153088, and its base is T or C;

[0041] Or the SNP site 86 is located in the white pheasant genome JADANK010001549.1:96570, and its base is G or A;

[0042] The reference genome of the SNP site is GCA_021292215.1.

[0043] In the process of studying the problem of individual identification of white pheasants, the inventors found that the identification methods based on genetic markers such as microsatellites proposed by previous researchers have the disadvantages of being prone to mutation or typing errors, while single nucleotide polymorphism (SNP) as a third-generation molecular marker has the advantages of being widely distributed in the genome, having a low mutation rate, and being easy to type and analyze compared with previous genetic markers such as microsatellites. Therefore, the inventors proposed to use at least 20 of the above-mentioned SNP sites as genetic markers, and use the genetic markers to perform individual identification of white pheasants. The individual identification results are accurate and reliable, and on the basis of the individual identification results, it is also possible to perform kinship, population genetic diversity, population genetic structure analysis, etc., to meet the protection research needs of the white pheasant species.

[0044] In one embodiment, the genetic markers include the following SNP sites:

[0045] The SNP site 1 is located in the white pheasant genome JADANK010000113.1:5324820, and its base is C or T;

[0046] The SNP site 2 is located in the white pheasant genome JADANK010000063.1:10039215, and its base is C or G;

[0047] The SNP site 3 is located in the white pheasant genome JADANK010000024.1:15904660, and its base is C or T;

[0048] The SNP site 4 is located in the white pheasant genome JADANK010001474.1:10085797, and its base is G or T;

[0049] The SNP site 5 is located in the white pheasant genome JADANK010001487.1:10047134, and its base is A or G;

[0050] The SNP site 6 is located in the white pheasant genome JADANK010001500.1:10081764, and its base is G or A;

[0051] The SNP site 7 is located in the white pheasant genome JADANK010001509.1:301199, and its base is C or T;

[0052] The SNP site 8 is located in the white pheasant genome JADANK010000146.1:13374829, and its base is A or G;

[0053] The SNP site 9 is located in the white pheasant genome JADANK010001524.1:10055431, and its base is G or A;

[0054] The SNP site 10 is located in the white pheasant genome JADANK010001544.1:10002213, and its base is T or C;

[0055] The SNP site 11 is located in the white pheasant genome JADANK010001485.1:5025375, and its base is A or G;

[0056] The SNP site 12 is located in the white pheasant genome JADANK010001510.1:10226541, and its base is G or C;

[0057] The SNP site 13 is located in the white pheasant genome JADANK010000033.1:5079977, and its base is T or C;

[0058] The SNP site 14 is located in the white pheasant genome JADANK010001522.1:5612065, and its base is G or A;

[0059] The SNP site 15 is located in the white pheasant genome JADANK010001502.1:10380701, and its base is A or G;

[0060] The SNP site 16 is located in the white pheasant genome JADANK010001472.1:10421851, and its base is A or G;

[0061] The SNP site 17 is located in the white pheasant genome JADANK010000316.1:17079, and its base is C or T;

[0062] The SNP site 18 is located in the white pheasant genome JADANK010001547.1:480922, and its base is A or G;

[0063] The SNP site 19 is located in the white pheasant genome JADANK010000006.1:10748586, and its base is T or C;

[0064] The SNP site 20 is located in the white pheasant genome JADANK010001468.1:10617303, and its base is G or A;

[0065] The SNP site 21 is located in the white pheasant genome JADANK010001469.1:10711749, and its base is C or T;

[0066] The SNP site 22 is located in the white pheasant genome JADANK010001482.1:5142234, and its base is A or G;

[0067] The SNP site 23 is located in the white pheasant genome JADANK010001511.1:5123818, and its base is C or T;

[0068] The SNP site 24 is located in the white pheasant genome JADANK010001470.1:5597501, and its base is A or G;

[0069] The SNP site 25 is located in the white pheasant genome JADANK010001521.1:5015305, and its base is C or T;

[0070] The SNP site 26 is located in the white pheasant genome JADANK010001492.1:261381, and its base is C or T;

[0071] The SNP site 27 is located in the white pheasant genome JADANK010001490.1:2096, and its base is G or T;

[0072] The SNP site 28 is located in the white pheasant genome JADANK010001471.1:5505053, and its base is C or T;

[0073] The SNP site 29 is located in the white pheasant genome JADANK010000102.1:6442137, and its base is A or G;

[0074] The SNP site 30 is located in the white pheasant genome JADANK010000003.1:823633, and its base is C or T;

[0075] The SNP site 31 is located in the white pheasant genome JADANK010000004.1:6106541, and its base is G or A;

[0076] The SNP site 32 is located in the white pheasant genome JADANK010001465.1:6256108, and its base is A or T;

[0077] The SNP site 33 is located in the white pheasant genome JADANK010000032.1:15474, and its base is G or A;

[0078] The SNP site 34 is located in the white pheasant genome JADANK010001506.1:5837633, and its base is T or C;

[0079] The SNP site 35 is located in the white pheasant genome JADANK010001466.1:6605868, and its base is C or T;

[0080] The SNP site 36 is located in the white pheasant genome JADANK010001467.1:7069787, and its base is C or T;

[0081] The SNP site 37 is located in the white pheasant genome JADANK010001489.1:637286, and its base is T or C;

[0082] The SNP site 38 is located in the white pheasant genome JADANK010001550.1:64201, and its base is G or A;

[0083] The SNP site 39 is located in the white pheasant genome JADANK010000001.1:111955, and its base is A or G;

[0084] The SNP site 40 is located in the white pheasant genome JADANK010000002.1:316812, and its base is G or C;

[0085] The SNP site 41 is located in the white pheasant genome JADANK010000005.1:73154, and its base is T or C;

[0086] The SNP site 42 is located in the white pheasant genome JADANK010000008.1:123053, and its base is T or C;

[0087] The SNP site 43 is located in the white pheasant genome JADANK010000019.1:138662, and its base is T or C;

[0088] The SNP site 44 is located in the white pheasant genome JADANK010000021.1:473931, and its base is T or C;

[0089] The SNP site 45 is located in the white pheasant genome JADANK010000022.1:151474, and its base is T or G;

[0090] The SNP site 46 is located in the white pheasant genome JADANK010000023.1:238776, and its base is C or T;

[0091] The SNP site 47 is located in the white pheasant genome JADANK010000029.1:1805, and its base is T or C;

[0092] The SNP site 48 is located in the white pheasant genome JADANK010000030.1:22918, and its base is C or T;

[0093] The SNP site 49 is located in the white pheasant genome JADANK010000031.1:38390, and its base is T or C;

[0094] The SNP site 50 is located in the white pheasant genome JADANK010000034.1:359, and its base is C or T;

[0095] The SNP site 51 is located in the white pheasant genome JADANK010000046.1:4067, and its base is G or A;

[0096] The SNP site 52 is located in the white pheasant genome JADANK010000048.1:74254, and its base is T or C;

[0097] The SNP site 53 is located in the white pheasant genome JADANK010000050.1:196194, and its base is T or C;

[0098] The SNP site 54 is located in the white pheasant genome JADANK010000061.1:1531940, and its base is A or G;

[0099] The SNP site 55 is located in the white pheasant genome JADANK010000062.1:212481, and its base is C or T;

[0100] The SNP site 56 is located in the white pheasant genome JADANK010000066.1:134569, and its base is G or A;

[0101] The SNP site 57 is located in the white pheasant genome JADANK010000067.1:133257, and its base is T or C;

[0102] The SNP site 58 is located in the white pheasant genome JADANK010000069.1:155575, and its base is T or C;

[0103] The SNP site 59 is located in the white pheasant genome JADANK010000070.1:65857, and its base is C or T;

[0104] The SNP site 60 is located in the white pheasant genome JADANK010000071.1:281355, and its base is A or G;

[0105] The SNP site 61 is located in the white pheasant genome JADANK010000073.1:110017, and its base is G or A;

[0106] The SNP site 62 is located in the white pheasant genome JADANK010000074.1:76097, and its base is A or G;

[0107] The SNP site 63 is located in the white pheasant genome JADANK010000086.1:167039, and its base is T or C;

[0108] The SNP site 64 is located in the white pheasant genome JADANK010000087.1:140185, and its base is C or T;

[0109] The SNP site 65 is located in the white pheasant genome JADANK010000091.1:582446, and its base is C or T;

[0110] The SNP site 66 is located in the white pheasant genome JADANK010000092.1:12529, and its base is T or C;

[0111] The SNP site 67 is located in the genome of the white pheasant JADANK010000100.1:20025, and its base is A or G;

[0112] The SNP site 68 is located in the white pheasant genome JADANK010000105.1:145727, and its base is G or A;

[0113] The SNP site 69 is located in the white pheasant genome JADANK010000106.1:32240, and its base is T or C;

[0114] The SNP site 70 is located in the white pheasant genome JADANK010000116.1:261398, and its base is C or T;

[0115] The SNP site 71 is located in the white pheasant genome JADANK010000144.1:54717, and its base is C or T;

[0116] The SNP site 72 is located in the white pheasant genome JADANK010001473.1:25594, and its base is A or G;

[0117] The SNP site 73 is located in the white pheasant genome JADANK010001486.1:111080, and its base is A or G;

[0118] The SNP site 74 is located in the white pheasant genome JADANK010001488.1:82694, and its base is A or G;

[0119] The SNP site 75 is located in the white pheasant genome JADANK010001491.1:73650, and its base is T or C;

[0120] The SNP site 76 is located in the white pheasant genome JADANK010001501.1:101470, and its base is A or G;

[0121] The SNP site 77 is located in the white pheasant genome JADANK010001504.1:66945, and its base is C or T;

[0122] The SNP site 78 is located in the white pheasant genome JADANK010001507.1:13120, and its base is C or T;

[0123] The SNP site 79 is located in the white pheasant genome JADANK010001526.1:275885, and its base is C or A;

[0124] The SNP site 80 is located in the white pheasant genome JADANK010001531.1:233119, and its base is T or C;

[0125] The SNP site 81 is located in the white pheasant genome JADANK010001533.1:1767861, and its base is T or C;

[0126] The SNP site 82 is located in the white pheasant genome JADANK010001534.1:134674, and its base is A or G;

[0127] The SNP site 83 is located in the white pheasant genome JADANK010001545.1:87524, and its base is G or A;

[0128] The SNP site 84 is located in the white pheasant genome JADANK010001546.1:84383, and its base is A or C;

[0129] The SNP site 85 is located in the white pheasant genome JADANK010001548.1:153088, and its base is T or C;

[0130] The SNP site 86 is located in the white pheasant genome JADANK010001549.1:96570, and its base is G or A.

[0131] The present invention also provides the application of the genetic marker in the individual identification of silver pheasants, wherein the individual identification includes: genotyping of varieties, and / or detection of similarity between any two silver pheasants.

[0132] The present invention also provides a primer set for detecting the genetic marker, characterized in that the primer set includes at least 20 of primers 1-86;

[0133] The primer 1 includes SEQ NO.1 and SEQ NO.2, the primer 2 includes SEQ NO.3 and SEQ NO.4, the primer 3 includes SEQ NO.5 and SEQ NO.6, the primer 4 includes SEQ NO.7 and SEQ NO.8, the primer 5 includes SEQ NO.9 and SEQ NO.10, the primer 6 includes SEQ NO.11 and SEQ NO.12, the primer 7 includes SEQ NO.13 and SEQ NO.14, the primer 8 includes SEQ NO.15 and SEQ NO.16, the primer 9 includes SEQ NO.17 and SEQ NO.18, the primer 10 includes SEQ NO.19 and SEQ NO.20, the primer 11 includes SEQ NO.21 and SEQ NO.22, the primer 12 includes SEQ NO.23 and SEQ NO.24, the primer 13 includes SEQ NO.25 and SEQ NO.26, the primer 14 includes SEQ NO.27 and SEQ NO.28, the primer 15 includes SEQ NO.29 and SEQ NO.30. NO.29 and SEQ NO.30, the primer 16 includes SEQ NO.31 and SEQ NO.32, the primer 17 includes SEQ NO.33 and SEQ NO.34, the primer 18 includes SEQ NO.35 and SEQ NO.36, the primer 19 includes SEQ NO.37 and SEQ NO.38, the primer 20 includes SEQ NO.39 and SEQ NO.40, the primer 21 includes SEQ NO.41 and SEQ NO.42, the primer 22 includes SEQ NO.43 and SEQ NO.44, the primer 23 includes SEQ NO.45 and SEQ NO.46, the primer 24 includes SEQ NO.47 and SEQ NO.48, the primer 25 includes SEQ NO.49 and SEQ NO.50, the primer 26 includes SEQ NO.51 and SEQ NO.52, the primer 27 includes SEQ NO.53 and SEQ NO.54, the primer 28 includes SEQ NO.55 and SEQ NO. NO.56, the primer 29 includes SEQ NO.57 and SEQ NO.58, the primer 30 includes SEQ NO.59 and SEQ NO.60, the primer 31 includes SEQ NO.61 and SEQ NO.62, the primer 32 includes SEQ NO.63 and SEQ NO.64, the primer 33 includes SEQ NO.65 and SEQ NO.66, the primer 34 includes SEQ NO.67 and SEQ NO.68, the primer 35 includes SEQ NO.69 and SEQ NO.70, the primer 36 includes SEQ NO.71 and SEQ NO.72, the primer 37 includes SEQ NO.73 and SEQ NO.74, the primer 38 includes SEQ NO.75 and SEQ NO.76, the primer 39 includes SEQ NO.77 and SEQ NO.78, the primer 40 includes SEQ NO.79 and SEQ NO.80, the primer 41 includes SEQ NO.81 and SEQ NO.82, the primer 42 includes SEQ NO.83 and SEQ NO.84, the primer 43 includes SEQ NO.85 and SEQ NO.86, the primer 44 includes SEQ NO.87 and SEQ NO.88, the primer 45 includes SEQ NO.89 and SEQ NO.90, the primer 46 includes SEQ NO.91 and SEQ NO.92, the primer 47 includes SEQ NO.93 and SEQ NO.94, the primer 48 includes SEQ NO.95 and SEQ NO.96, the primer 49 includes SEQ NO.97 and SEQ NO.98, the primer 50 includes SEQ NO.99 and SEQ NO. NO.100, the primer 51 includes SEQ NO.101 and SEQ NO.102, the primer 52 includes SEQ NO.103 and SEQ NO.104, the primer 53 includes SEQ NO.105 and SEQ NO.106, the primer 54 includes SEQ NO.107 and SEQ NO.108, the primer 55 includes SEQ NO.109 and SEQ NO.110, the primer 56 includes SEQ NO.111 and SEQ NO.112, the primer 57 includes SEQ NO.113 and SEQ NO.114, the primer 58 includes SEQ NO.115 and SEQ NO.116, the primer 59 includes SEQ NO.117 and SEQ NO.118, the primer 60 includes SEQ NO.119 and SEQ NO.120, the primer 61 includes SEQ NO.121 and SEQ NO.122, the primer 62 includes SEQ NO.123 and SEQ NO.124. NO.124, the primer 63 includes SEQ NO.125 and SEQ NO.126, the primer 64 includes SEQ NO.127 and SEQ NO.128, the primer 65 includes SEQ NO.129 and SEQ NO.130, the primer 66 includes SEQ NO.131 and SEQ NO.132, the primer 67 includes SEQ NO.133 and SEQ NO.134, the primer 68 includes SEQ NO.135 and SEQ NO.136, the primer 69 includes SEQ NO.137 and SEQ NO.138, the primer 70 includes SEQ NO.139 and SEQ NO.140, the primer 71 includes SEQ NO.141 and SEQ NO.142, the primer 72 includes SEQ NO.143 and SEQ NO.144, the primer 73 includes SEQ NO.145 and SEQ NO.146, the primer 74 includes SEQ NO.147 and SEQ NO.148, the primer 75 includes SEQ NO.149 and SEQ NO.150, the primer 76 includes SEQ NO.151 and SEQ NO.152, the primer 77 includes SEQ NO.153 and SEQ NO.154, the primer 78 includes SEQ NO.155 and SEQ NO.156, the primer 79 includes SEQ NO.157 and SEQ NO.158, the primer 80 includes SEQ NO.159 and SEQ NO.160, the primer 81 includes SEQ NO.161 and SEQ NO.162, the primer 82 includes SEQ NO.163 and SEQ NO.164, the primer 83 includes SEQ NO.165 and SEQ NO.166 No. 166, the primer 84 includes SEQ No. 167 and SEQ No. 168, the primer 85 includes SEQ No. 169 and SEQ No. 170, and the primer 86 includes SEQ No. 171 and SEQ No. 172.

[0134] In one embodiment, the primer set includes primers 1-86.

[0135] The present invention also provides a kit for identifying silver pheasant individuals, the kit comprising a PCR system, and the PCR system comprising the primer set.

[0136] In one embodiment, the PCR system includes a first PCR and a second PCR, the first PCR is a multiplex PCR, and the second PCR is a multiplex PCR.

[0137] In one embodiment, the reaction conditions of the first PCR are: 94-96°C, 13-17 min; 93-95°C, 28-32 s, 58-62°C, 8-12 min, 70-74°C, 28-32 s, 4-6 cycles; 92-96°C, 28-32 s, 58-62°C, 0.8-1.2 min, 70-74°C, 28-32 s, 18-22 cycles.

[0138] In one embodiment, the reaction conditions of the second PCR are: 94-96°C, 13-17 min; 93-95°C, 28-32 s, 58-62°C, 2-6 min, 70-74°C, 28-32 s, 4-6 cycles; 92-96°C, 28-32 s, 58-62°C, 0.8-1.2 min, 70-74°C, 28-32 s, 8-12 cycles.

[0139] The present invention also provides application of the reagent in individual identification of white pheasants, wherein the individual identification includes: gene typing of the white pheasants, analysis of the kinship of the white pheasants, and analysis of the genetic diversity of the white pheasant population.

[0140] The present invention also provides a method for identifying silver pheasants individually, comprising the following steps: extracting DNA of a sample to be tested, performing PCR amplification using the kit to obtain a PCR amplification product, performing high-throughput sequencing to obtain genotype data of the genetic marker, and performing individual identification according to the genotype data.

[0141] In one embodiment, the sample to be tested includes feathers and / or blood.

[0142] Compared with the prior art, the present invention has the following beneficial effects:

[0143] The invention discloses a genetic marker for individual identification of silver pheasants. The genetic marker is used to perform individual identification of silver pheasants. The individual identification result is accurate and reliable. Moreover, on the basis of the individual identification result, kinship, population genetic diversity, group genetic structure analysis and the like can be performed, thereby meeting the protection research requirements of the silver pheasant species. DETAILED DESCRIPTION

[0144] The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0145] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0146] source:

[0147] Unless otherwise specified, the reagents, materials, and equipment used in this example are all commercially available; the experimental methods are all conventional experimental methods in the art unless otherwise specified.

[0148] Example

[0149] 1. Selection of SNP sites.

[0150] 1. SNP dataset detection.

[0151] DNA was extracted from blood samples of 13 silver pheasants. After constructing DNA libraries, paired-end 150bp sequencing was performed on the Illumina NovaSeq 6000 platform. The depth of whole genome resequencing for each individual was more than 10Gb. After quality control, the raw reads data were aligned to the reference genome of the silver pheasant (accession number GCA_021292215.1 in NCBI) using BWA software, then converted into bam files using Samtools software, and duplicate reads were removed using Picard software. Finally, the HaplotypeCaller and GenotypeGVCFs commands in the Genome Analysis ToolKit software were used for SNP detection, and then the VariantFiltration command was used for quality control. The conditions were DP<59; DP>534; QD<2.0; FS>60.0; MQ<40.0; MQRankSum<-12.5; ReadPosRankSum<-8.0; SOR>3.0, ultimately forming a reliable reference population of white pheasant SNP data set.

[0152] 2. SNP site screening.

[0153] We further filtered the SNP dataset of the white pheasant to select SNP sites suitable for individual identification. The screening criteria are as follows: (1) only select scaffolds with a length greater than 1Mb in the reference genome; (2) the minimum allele frequency is greater than 0.4; (4) select SNP sites located in the coding region; (2) the physical distance between two SNP sites on the genome is greater than 5Mb; (3) there are no insertions or deletions within 500bp on both sides of the SNP site. According to the above conditions, 200 SNP sites were selected, and then the 500bp sequences on both sides of these SNP sites were blasted to the reference genome of the white pheasant, and only SNP sites with a highly homologous result of more than 95% were retained. Finally, primers were designed and evaluated for the sequences of these SNP sites, and finally 86 SNP sites were selected, which were located on 86 scaffolds. The specific information is shown in Table 1:

[0154] Table 1 Information on 86 SNP loci in Silver Pheasant

[0155]

[0156]

[0157]

[0158] 2. SNP locus typing.

[0159] 1. Extract sample DNA from the validation population.

[0160] 25 samples of silver pheasant were randomly collected, including 20 blood samples and 5 feather samples, 2 of which were from the same individual. DNA was extracted using GENFINE's nucleic acid extraction reagent (magnetic bead method).

[0161] 2. Amplification of SNP sites.

[0162] The multiplex PCR method was used to amplify the above 86 SNP sites. The primer information is shown in Table 2.

[0163] The multiplex PCR amplification system was as follows: 2 μL DNA, 1 μL Buffer (10×), 2 μL premixed primer combination (50 nM), 0.8 μL dNTP (2.5 mM), 0.1 μL enzyme (5 U / μL), 1 μL Mg 2+ (100 mM), 3.2 μL ddH2O, and 10 μL paraffin oil.

[0164] The PCR program was: 95°C, 15 min; 94°C, 30 s, 60°C, 10 min, 72°C, 30 s, 4 cycles; 94°C, 30 s, 60°C, 1 min, 72°C, 30 s, 20 cycles.

[0165] The second round of PCR was performed using the first round PCR product as a template. In order to meet the needs of subsequent high-throughput sequencing, adapters (Shanghai Yihe Applied Biotechnology Co., Ltd.) were added to each sample for the second round of PCR.

[0166] 100 μL ddH2O was added to the multiplex PCR products of each sample in the first round, centrifuged instantly, and allowed to stand at room temperature for 10 min. The dilution was then used as the template for the second round of PCR. The amplification reaction system was as follows: 10 μL template, 2 μL Buffer (10×), 3.6 μL Barcode (2 μM), 0.8 μL dNTP (2.5 mM), 0.1 μL enzyme (5 U / μL), 1 μL Mg 2+ (100 mM), 3.6 μL ddH2O, and 20 μL paraffin oil.

[0167] The PCR program was as follows: 95°C, 15 min; 94°C, 30 s, 60°C, 4 min, 72°C, 30 s, 5 cycles; 94°C, 30 s, 60°C, 1 min, 72°C, 30 s, 10 cycles.

[0168] Table 2 Primer information for amplification of 86 SNP loci in Siberian pheasant

[0169]

[0170]

[0171]

[0172]

[0173]

[0174] 3. High-throughput sequencing and genotyping.

[0175] 5 μL of the final PCR product of each sample was mixed, and the library was mixed and shaken overnight. Then, the mixed library was electrophoresed, the target fragment was cut and recovered, and the purified PCR product was accurately quantified and diluted to the concentration required for high-throughput sequencing. The PCR product mixed library was sequenced on the Illumina X-ten sequencing platform, and the operation process was carried out according to the standard SOP. The raw data obtained by sequencing was converted into sequence data by Illumina bcl2fastq software and stored in fastq file format. Illumina bcl2fastq was used to count the number of reads amplified from the SNP site of each sample according to the index information of each sample. Illumina RTA software was used for image recognition and base determination, and finally the genotype of each SNP site of each sample was obtained.

[0176] Through statistical analysis, the average sequencing depth of 86 SNP sites in the 25 validation samples reached 5499×, indicating that the reliability of genotyping of SNP sites is very high. The validation samples were divided into blood and feathers with feather follicles that had just grown. The results showed that both samples could be used for this method. The feathers with feather follicles were collected without trauma, conveniently and more practically.

[0177] The genotyping results of the 86 SNP loci in this group found that the typing rate of the SNP loci of all individuals reached 100%, all of which were biallelic, and the average polymorphic information content PIC value was 0.3469. Among them, the allele frequency of 60 SNP loci was around 0.4, and 69 SNP loci were in Hardy-Weinberg equilibrium, indicating that this group of SNP loci has good individual identification and population genetic diversity assessment for white pheasants.

[0178] Table 3 Genotyping statistics of 86 SNP loci

[0179]

[0180]

[0181]

[0182] Note: ** in the above table means no display.

[0183] 4. Individual identification analysis.

[0184] The genotyping data of 86 SNP sites of 25 samples were sorted into the recognition format of Cervus3.0.7 software, and the Identity Analysis program in the Analysis module of the software was used for individual identification analysis. The results showed that among the 25 individuals, the genotypes of the 86 SNP sites of the two repeated samples were all the same and were marked as "Exact match", that is, they were identified as the same individual, indicating that this method is reliable and reproducible. The other individuals were all shown as "Excluded" between each other, indicating that the group of 86 sites can accurately identify individuals.

[0185] 3. Application of SNP sites. Among the 86 SNP sites in this embodiment, 3 groups of SNP site genotyping data were randomly selected, with 20 SNP sites in each group, to verify the effect of SNP sites in individual identification. The specific groups are as follows:

[0186] The first group selected (SNP site 1, SNP site 5, SNP site 9, SNP site 13, SNP site 17, SNP site 21, SNP site 25, SNP site 29, SNP site 33, SNP site 37, SNP site 41, SNP site 45, SNP site 49, SNP site 53, SNP site 57, SNP site 61, SNP site 65, SNP site 69, SNP site 73, SNP site 77);

[0187] The second group selected (SNP site 2, SNP site 6, SNP site 10, SNP site 14, SNP site 18, SNP site 22, SNP site 26, SNP site 30, SNP site 34, SNP site 38, SNP site 42, SNP site 46, SNP site 50, SNP site 54, SNP site 58, SNP site 62, SNP site 66, SNP site 70, SNP site 74, SNP site 78);

[0188] The third group selection (SNP site 3, SNP site 7, SNP site 11, SNP site 15, SNP site 19, SNP site 23, SNP site 27, SNP site 31, SNP site 35, SNP site 39, SNP site 43, SNP site 47, SNP site 51, SNP site 55, SNP site 59, SNP site 63, SNP site 67, SNP site 71, SNP site 75, SNP site 79).

[0189] Cervus3.0.7 software was used to perform individual identification on the above three groups of SNP site data. The analysis results of the three groups of SNP sites for two repeated samples all showed that 20 SNPs were completely matched, and the pID values ​​were 9.49E-12, 6.90E-11 and 2.83E-11, respectively, and the judgment results were all "Exact match". For the other 24 individuals, the number of SNP matches between two individuals was 2-15, 2-18, and 3-16 in these three groups, respectively, and the pID values ​​were all 0, and the judgment results were all "Excluded", indicating that the use of at least 20 SNP sites among the 86 SNP sites in this embodiment, the genotyping data detected can all be used for accurate individual identification, and have the same effect as using 86 SNP sites.

[0190] Therefore, the number of SNP sites can be selected according to the number of individuals to be identified. According to the 86 SNP sites provided by the present invention, since each SNP site is a double allele, three genotypes can be formed, forming an 86-bit digital label, which can theoretically give 3 86 The molecular tags are provided for each individual white pheasant to meet the needs of individual identification and tracking of the white pheasant population.

[0191] This group of 86 SNP sites can be used to effectively identify individuals of white pheasants, and can also be applied to subsequent kinship and genetic diversity assessments.

[0192] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0193] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A primer set for detecting a genetic marker, characterized in that: The genetic marker includes the following 86 SNP sites, and the reference genome of the SNP sites is GCA_021292215.1: The SNP site 1 is located in the white pheasant genome JADANK010000113.1:5324820, and its base is C or T; The SNP site 2 is located in the white pheasant genome JADANK010000063.1:10039215, and its base is C or G; The SNP site 3 is located in the white pheasant genome JADANK010000024.1:15904660, and its base is C or T; The SNP site 4 is located in the white pheasant genome JADANK010001474.1:10085797, and its base is G or T; The SNP site 5 is located in the white pheasant genome JADANK010001487.1:10047134, and its base is A or G; The SNP site 6 is located in the white pheasant genome JADANK010001500.1:10081764, and its base is G or A; The SNP site 7 is located in the white pheasant genome JADANK010001509.1:301199, and its base is C or T; The SNP site 8 is located in the white pheasant genome JADANK010000146.1:13374829, and its base is A or G; The SNP site 9 is located in the white pheasant genome JADANK010001524.1:10055431, and its base is G or A; The SNP site 10 is located in the white pheasant genome JADANK010001544.1:10002213, and its base is T or C; The SNP site 11 is located in the white pheasant genome JADANK010001485.1:5025375, and its base is A or G; The SNP site 12 is located in the white pheasant genome JADANK010001510.1:10226541, and its base is G or C; The SNP site 13 is located in the white pheasant genome JADANK010000033.1:5079977, and its base is T or C; The SNP site 14 is located in the white pheasant genome JADANK010001522.1:5612065, and its base is G or A; The SNP site 15 is located in the white pheasant genome JADANK010001502.1:10380701, and its base is A or G; The SNP site 16 is located in the white pheasant genome JADANK010001472.1:10421851, and its base is A or G; The SNP site 17 is located in the white pheasant genome JADANK010000316.1:17079, and its base is C or T; The SNP site 18 is located in the white pheasant genome JADANK010001547.1:480922, and its base is A or G; The SNP site 19 is located in the white pheasant genome JADANK010000006.1:10748586, and its base is T or C; The SNP site 20 is located in the white pheasant genome JADANK010001468.1:10617303, and its base is G or A; The SNP site 21 is located in the white pheasant genome JADANK010001469.1:10711749, and its base is C or T; The SNP site 22 is located in the white pheasant genome JADANK010001482.1:5142234, and its base is A or G; The SNP site 23 is located in the white pheasant genome JADANK010001511.1:5123818, and its base is C or T; The SNP site 24 is located in the white pheasant genome JADANK010001470.1:5597501, and its base is A or G; The SNP site 25 is located in the white pheasant genome JADANK010001521.1:5015305, and its base is C or T; The SNP site 26 is located in the white pheasant genome JADANK010001492.1:261381, and its base is C or T; The SNP site 27 is located in the white pheasant genome JADANK010001490.1:2096, and its base is G or T; The SNP site 28 is located in the white pheasant genome JADANK010001471.1:5505053, and its base is C or T; The SNP site 29 is located in the white pheasant genome JADANK010000102.1:6442137, and its base is A or G; The SNP site 30 is located in the white pheasant genome JADANK010000003.1:823633, and its base is C or T; The SNP site 31 is located in the white pheasant genome JADANK010000004.1:6106541, and its base is G or A; The SNP site 32 is located in the white pheasant genome JADANK010001465.1:6256108, and its base is A or T; The SNP site 33 is located in the white pheasant genome JADANK010000032.1:15474, and its base is G or A; The SNP site 34 is located in the white pheasant genome JADANK010001506.1:5837633, and its base is T or C; The SNP site 35 is located in the white pheasant genome JADANK010001466.1:6605868, and its base is C or T; The SNP site 36 is located in the white pheasant genome JADANK010001467.1:7069787, and its base is C or T; The SNP site 37 is located in the white pheasant genome JADANK010001489.1:637286, and its base is T or C; The SNP site 38 is located in the white pheasant genome JADANK010001550.1:64201, and its base is G or A; The SNP site 39 is located in the white pheasant genome JADANK010000001.1:111955, and its base is A or G; The SNP site 40 is located in the white pheasant genome JADANK010000002.1:316812, and its base is G or C; The SNP site 41 is located in the white pheasant genome JADANK010000005.1:73154, and its base is T or C; The SNP site 42 is located in the white pheasant genome JADANK010000008.1:123053, and its base is T or C; The SNP site 43 is located in the white pheasant genome JADANK010000019.1:138662, and its base is T or C; The SNP site 44 is located in the white pheasant genome JADANK010000021.1:473931, and its base is T or C; The SNP site 45 is located in the white pheasant genome JADANK010000022.1:151474, and its base is T or G; The SNP site 46 is located in the white pheasant genome JADANK010000023.1:238776, and its base is C or T; The SNP site 47 is located in the white pheasant genome JADANK010000029.1:1805, and its base is T or C; The SNP site 48 is located in the white pheasant genome JADANK010000030.1:22918, and its base is C or T; The SNP site 49 is located in the white pheasant genome JADANK010000031.1:38390, and its base is T or C; The SNP site 50 is located in the white pheasant genome JADANK010000034.1:359, and its base is C or T; The SNP site 51 is located in the white pheasant genome JADANK010000046.1:4067, and its base is G or A; The SNP site 52 is located in the white pheasant genome JADANK010000048.1:74254, and its base is T or C; The SNP site 53 is located in the white pheasant genome JADANK010000050.1:196194, and its base is T or C; The SNP site 54 is located in the white pheasant genome JADANK010000061.1:1531940, and its base is A or G; The SNP site 55 is located in the white pheasant genome JADANK010000062.1:212481, and its base is C or T; The SNP site 56 is located in the white pheasant genome JADANK010000066.1:134569, and its base is G or A; The SNP site 57 is located in the white pheasant genome JADANK010000067.1:133257, and its base is T or C; The SNP site 58 is located in the white pheasant genome JADANK010000069.1:155575, and its base is T or C; The SNP site 59 is located in the white pheasant genome JADANK010000070.1:65857, and its base is C or T; The SNP site 60 is located in the white pheasant genome JADANK010000071.1:281355, and its base is A or G; The SNP site 61 is located in the white pheasant genome JADANK010000073.1:110017, and its base is G or A; The SNP site 62 is located in the white pheasant genome JADANK010000074.1:76097, and its base is A or G; The SNP site 63 is located in the white pheasant genome JADANK010000086.1:167039, and its base is T or C; The SNP site 64 is located in the white pheasant genome JADANK010000087.1:140185, and its base is C or T; The SNP site 65 is located in the white pheasant genome JADANK010000091.1:582446, and its base is C or T; The SNP site 66 is located in the white pheasant genome JADANK010000092.1:12529, and its base is T or C; The SNP site 67 is located in the genome of the white pheasant JADANK010000100.1:20025, and its base is A or G; The SNP site 68 is located in the white pheasant genome JADANK010000105.1:145727, and its base is G or A; The SNP site 69 is located in the white pheasant genome JADANK010000106.1:32240, and its base is T or C; The SNP site 70 is located in the white pheasant genome JADANK010000116.1:261398, and its base is C or T; The SNP site 71 is located in the white pheasant genome JADANK010000144.1:54717, and its base is C or T; The SNP site 72 is located in the white pheasant genome JADANK010001473.1:25594, and its base is A or G; The SNP site 73 is located in the white pheasant genome JADANK010001486.1:111080, and its base is A or G; The SNP site 74 is located in the white pheasant genome JADANK010001488.1:82694, and its base is A or G; The SNP site 75 is located in the white pheasant genome JADANK010001491.1:73650, and its base is T or C; The SNP site 76 is located in the white pheasant genome JADANK010001501.1:101470, and its base is A or G; The SNP site 77 is located in the white pheasant genome JADANK010001504.1:66945, and its base is C or T; The SNP site 78 is located in the white pheasant genome JADANK010001507.1:13120, and its base is C or T; The SNP site 79 is located in the white pheasant genome JADANK010001526.1:275885, and its base is C or A; The SNP site 80 is located in the white pheasant genome JADANK010001531.1:233119, and its base is T or C; The SNP site 81 is located in the white pheasant genome JADANK010001533.1:1767861, and its base is T or C; The SNP site 82 is located in the white pheasant genome JADANK010001534.1:134674, and its base is A or G; The SNP site 83 is located in the white pheasant genome JADANK010001545.1:87524, and its base is G or A; The SNP site 84 is located in the white pheasant genome JADANK010001546.1:84383, and its base is A or C; The SNP site 85 is located in the white pheasant genome JADANK010001548.1:153088, and its base is T or C; The SNP site 86 is located in the white pheasant genome JADANK010001549.1:96570, and its base is G or A; The primer set includes primers 1-86; The primer 1 includes SEQ NO.1 and SEQ NO.2, the primer 2 includes SEQ NO.3 and SEQ NO.4, the primer 3 includes SEQ NO.5 and SEQ NO.6, the primer 4 includes SEQ NO.7 and SEQ NO.8, the primer 5 includes SEQ NO.9 and SEQ NO.10, the primer 6 includes SEQ NO.11 and SEQ NO.12, the primer 7 includes SEQ NO.13 and SEQ NO.14, the primer 8 includes SEQ NO.15 and SEQ NO.16, the primer 9 includes SEQ NO.17 and SEQ NO.18, the primer 10 includes SEQ NO.19 and SEQ NO.20, the primer 11 includes SEQ NO.21 and SEQ NO.22, the primer 12 includes SEQ NO.23 and SEQ NO.24, the primer 13 includes SEQ NO.25 and SEQ NO.26, the primer 14 includes SEQ NO.27 and SEQ NO.28, the primer 15 includes SEQ NO.29 and SEQ NO.

30. NO.29 and SEQ NO.30, the primer 16 includes SEQ NO.31 and SEQ NO.32, the primer 17 includes SEQ NO.33 and SEQ NO.34, the primer 18 includes SEQ NO.35 and SEQ NO.36, the primer 19 includes SEQ NO.37 and SEQ NO.38, the primer 20 includes SEQ NO.39 and SEQ NO.40, the primer 21 includes SEQ NO.41 and SEQ NO.42, the primer 22 includes SEQ NO.43 and SEQ NO.44, the primer 23 includes SEQ NO.45 and SEQ NO.46, the primer 24 includes SEQ NO.47 and SEQ NO.48, the primer 25 includes SEQ NO.49 and SEQ NO.50, the primer 26 includes SEQ NO.51 and SEQ NO.52, the primer 27 includes SEQ NO.53 and SEQ NO.54, the primer 28 includes SEQ NO.55 and SEQ NO. NO.56, the primer 29 includes SEQ NO.57 and SEQ NO.58, the primer 30 includes SEQ NO.59 and SEQ NO.60, the primer 31 includes SEQ NO.61 and SEQ NO.62, the primer 32 includes SEQ NO.63 and SEQ NO.64, the primer 33 includes SEQ NO.65 and SEQ NO.66, the primer 34 includes SEQ NO.67 and SEQ NO.68, the primer 35 includes SEQ NO.69 and SEQ NO.70, the primer 36 includes SEQ NO.71 and SEQ NO.72, the primer 37 includes SEQ NO.73 and SEQ NO.74, the primer 38 includes SEQ NO.75 and SEQ NO.76, the primer 39 includes SEQ NO.77 and SEQ NO.78, the primer 40 includes SEQ NO.79 and SEQ NO.80, the primer 41 includes SEQ NO.81 and SEQ NO.82, the primer 42 includes SEQ NO.83 and SEQ NO.84, the primer 43 includes SEQ NO.85 and SEQ NO.86, the primer 44 includes SEQ NO.87 and SEQ NO.88, the primer 45 includes SEQ NO.89 and SEQ NO.90, the primer 46 includes SEQ NO.91 and SEQ NO.92, the primer 47 includes SEQ NO.93 and SEQ NO.94, the primer 48 includes SEQ NO.95 and SEQ NO.96, the primer 49 includes SEQ NO.97 and SEQ NO.98, the primer 50 includes SEQ NO.99 and SEQ NO. NO.100, the primer 51 includes SEQ NO.101 and SEQ NO.102, the primer 52 includes SEQ NO.103 and SEQ NO.104, the primer 53 includes SEQ NO.105 and SEQ NO.106, the primer 54 includes SEQ NO.107 and SEQ NO.108, the primer 55 includes SEQ NO.109 and SEQ NO.110, the primer 56 includes SEQ NO.111 and SEQ NO.112, the primer 57 includes SEQ NO.113 and SEQ NO.114, the primer 58 includes SEQ NO.115 and SEQ NO.116, the primer 59 includes SEQ NO.117 and SEQ NO.118, the primer 60 includes SEQ NO.119 and SEQ NO.120, the primer 61 includes SEQ NO.121 and SEQ NO.122, the primer 62 includes SEQ NO.123 and SEQ NO.

124. NO.124, the primer 63 includes SEQ NO.125 and SEQ NO.126, the primer 64 includes SEQ NO.127 and SEQ NO.128, the primer 65 includes SEQ NO.129 and SEQ NO.130, the primer 66 includes SEQ NO.131 and SEQ NO.132, the primer 67 includes SEQ NO.133 and SEQ NO.134, the primer 68 includes SEQ NO.135 and SEQ NO.136, the primer 69 includes SEQ NO.137 and SEQ NO.138, the primer 70 includes SEQ NO.139 and SEQ NO.140, the primer 71 includes SEQ NO.141 and SEQ NO.142, the primer 72 includes SEQ NO.143 and SEQ NO.144, the primer 73 includes SEQ NO.145 and SEQ NO.146, the primer 74 includes SEQ NO.147 and SEQ NO.148, the primer 75 includes SEQ NO.149 and SEQ NO.150, the primer 76 includes SEQ NO.151 and SEQ NO.152, the primer 77 includes SEQ NO.153 and SEQ NO.154, the primer 78 includes SEQ NO.155 and SEQ NO.156, the primer 79 includes SEQ NO.157 and SEQ NO.158, the primer 80 includes SEQ NO.159 and SEQ NO.160, the primer 81 includes SEQ NO.161 and SEQ NO.162, the primer 82 includes SEQ NO.163 and SEQ NO.164, the primer 83 includes SEQ NO.165 and SEQ NO.166 No. 166, the primer 84 includes SEQ No. 167 and SEQ No. 168, the primer 85 includes SEQ No. 169 and SEQ No. 170, and the primer 86 includes SEQ No. 171 and SEQ No.

172.

2. A kit for identifying individual silver pheasants, characterized in that: The kit comprises a PCR system, and the PCR system comprises the primer set according to claim 1.

3. Use of the kit according to claim 2 in individual identification of silver pheasant, wherein the individual identification comprises: Genotyping of the white pheasant, analysis of the white pheasant's genetic relationship, and analysis of the genetic diversity of the white pheasant population.

4. A method for identifying individual silver pheasants, characterized in that: The following steps are involved: Extract DNA from the sample to be tested, perform PCR amplification using the kit described in claim 2 to obtain PCR amplification products, perform high-throughput sequencing to obtain genotype data of the genetic markers of the primer set described in claim 1, and perform individual identification based on the genotype data.

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