SNP molecular markers for identification of oil-tea camellia varieties and their applications

Abstract

The present invention relates to the technical field of plant molecular markers, and in particular to SNP molecular markers for identifying oil-tea camellia varieties and their applications. The 31 core SNP molecular markers for identifying oil-tea camellia variety resources provided by the present invention can accurately distinguish all tested oil-tea camellia variety resources, and provide effective molecular tools and methods for oil-tea camellia variety identification, variety tracing, and authenticity and purity detection of seedlings. The DNA fingerprint of oil-tea camellia variety resources constructed by these SNP molecular markers is combined with basic attribute information such as the origin, resource type, and subpopulation distribution of the variety resources to construct a unique molecular identity card for each oil-tea camellia variety resource, and generate corresponding barcodes and QR codes, providing technical support for rapid detection of oil-tea camellia variety quality.

Description

Technical Field

[0001] The invention relates to the technical field of plant molecular markers, and in particular to SNP molecular markers for identifying oil-tea camellia varieties and applications thereof. Background Art

[0002] Camellia oleifera Camellia oleifera ), as a woody edible oil crop, has been widely cultivated in southern China for more than 2,300 years. In recent years, the oil-tea industry has developed rapidly and has become one of the four major oil crops in my country. For the healthy development of the oil-tea industry, good varieties and seedlings are the basis and key. In recent years, with the systematic and in-depth breeding work, high-yield and high-quality national and provincial oil-tea varieties have continued to emerge. However, under the background of rapid industrial development and huge demand for oil-tea varieties, the same variety has been approved (recognized) in different provinces, resulting in "same thing with different names", and "same name but different things" caused by inferior seedlings. Because the phenotypic traits are greatly affected by the environment, and the phenotypic differences of most high-yield oil-tea varieties are small, it is urgent to establish a set of common oil-tea variety identification methods with simple operation and accurate identification results.

[0003] Molecular markers can directly detect differences at the DNA level, are independent of phenotypic traits, have strong stability and good repeatability, and have been widely used in plant molecular fingerprinting, molecular ID cards, and variety identification research. However, due to the large genome and complex ploidy, the research progress of oil-tea camellia molecular marker technology is extremely slow. In recent years, with the development of a new generation of high-throughput sequencing technology, oil-tea camellia molecular marker technology has made great progress. Recently, the common oil-tea camellia ( C. oleifera var. nanyongensis )、Zhejiang safflower tea( C. chekiangoleosa )、Angustifolia oleifera( C. lanceoleosa The publication of the genomes of three oil-Camellia species, including the first and second varieties, has provided a good platform for exploring and developing polymorphic markers within the entire genome of oil-Camellia. Lin et al. (Lin P, Wang K, Wang Y, et al. The genome of oil-Camellia and population genomics analysis provide insights into seed oil domestication. Genome Biology, 2022, 23:14.) detected SNP markers in the transcriptome of common oil-Camellia based on high-throughput sequencing, confirming the presence of abundant single nucleotide polymorphisms in the common oil-Camellia population, indicating that it is feasible to use SNPs to construct a fingerprint map of common oil-Camellia varieties.

[0004] Establishing a SNP molecular marker database of Camellia oleifera variety resources, screening important SNP sites to develop DNA fingerprint maps of Camellia oleifera variety resources, and constructing a molecular identity card for Camellia oleifera variety resources are of great significance for identification and traceability of Camellia oleifera varieties. Summary of the invention

[0005] The purpose of the present invention is to provide SNP molecular markers for identifying oil-tea camellia varieties and applications thereof.

[0006] The present invention utilizes the SNP sites in the transcriptome of 221 common Camellia oleifera clones and adopts a series of strict filtering parameters to screen out polymorphism information content ( PIC ) high, linkage disequilibrium decay distance ( LD ) to accurately distinguish the core SNP markers of each asexual lineage, and to establish a molecular fingerprint and molecular identity card specific to the asexual lineage, in order to provide methods and basis for the identification and traceability of common tea oil varieties.

[0007] Specifically, 221 common oil-tea camellia varieties were used as materials to extract immature seed RNA and perform transcriptome sequencing. Using the diploid Nanrong oil-tea camellia genome as a reference, the SNP loci of the tested oil-tea camellia varieties were identified and genotyped, and the integrity of the SNP typing data in the population, the distribution on the genome, the SNP polymorphism information content and the minimum allele frequency ( MAF ) and other information, screened the core SNP sites and verified them by Sanger sequencing. After obtaining the optimal SNP site combination, the molecular identity card of oil-tea camellia variety resources was constructed in combination with the basic information of variety resources. Using the above method, a total of 1,849,953 high-quality SNP sites were detected from the oil-tea camellia transcriptome. The analysis found that the SNP of the oil-tea camellia population PIC is 0.2073, MAF is 0.1648. PIC , LD Parameters such as PCR were used to screen out 31 core sites with high polymorphism from all SNP sites, and Sanger sequencing verified that the accuracy of genotyping of 8 core sites was above 91.36%. The core sites were used to form a DNA fingerprint map, which could distinguish all the participating Camellia oleifera variety resources. A 66-digit molecular ID card of Camellia oleifera variety resources was constructed by combining the DNA fingerprint map with the basic information of Camellia oleifera variety resources.

[0008] Specifically, the present invention provides the following technical solutions:

[0009] In a first aspect, the present invention provides SNP molecular markers for identifying oil-tea camellia varieties, wherein the SNP molecular markers include any one or more combinations of the following SNP molecular markers:

[0010] SNP-1-16341109, which contains a nucleotide sequence with a polymorphism of A / G at position 16341109 of chromosome 1 of Camellia oleifera;

[0011] SNP-1-22551920, which contains a nucleotide sequence with a polymorphism of C / T at position 22551920 of chromosome 1 of Camellia oleifera;

[0012] SNP-1-34038354, which contains a nucleotide sequence with a polymorphism of T / A at position 34038354 of chromosome 1 of Camellia oleifera;

[0013] SNP-1-49052505, which contains a nucleotide sequence with a polymorphism of A / G at position 49052505 of chromosome 1 of Camellia oleifera;

[0014] SNP-1-110833857, which contains a nucleotide sequence with a polymorphism of G / C at position 110833857 of chromosome 1 of Camellia oleifera;

[0015] SNP-1-115890029, which contains a nucleotide sequence with a polymorphism of A / G at position 115890029 of chromosome 1 of Camellia oleifera;

[0016] SNP-1-188555309, which contains a nucleotide sequence with a polymorphism of T / G at position 188555309 of chromosome 1 of Camellia oleifera;

[0017] SNP-2-4697104, which contains a nucleotide sequence with a polymorphism of T / A at position 4697104 on chromosome 2 of Camellia oleifera;

[0018] SNP-2-102046792, which contains a nucleotide sequence with a polymorphism of C / A at position 102046792 of chromosome 2 of Camellia oleifera;

[0019] SNP-3-58564519, which contains a nucleotide sequence with a polymorphism of C / T at position 58564519 of chromosome 3 of Camellia oleifera;

[0020] SNP-4-5589980, which contains a nucleotide sequence with a polymorphism of T / G at position 5589980 of chromosome 4 of Camellia oleifera;

[0021] SNP-5-59238652, which contains a nucleotide sequence with a polymorphism of G / C at position 59238652 of chromosome 5 of Camellia oleifera;

[0022] SNP-5-135461353, which contains a nucleotide sequence with a polymorphism of T / C at position 135461353 of chromosome 5 of Camellia oleifera;

[0023] SNP-5-175443498, which contains a nucleotide sequence with a polymorphism of T / G located at position 175443498 of chromosome 5 of Camellia oleifera;

[0024] SNP-6-123202382, which contains a nucleotide sequence with a polymorphism of A / C at position 123202382 of chromosome 6 of Camellia oleifera;

[0025] SNP-6-125991240, which contains a nucleotide sequence with a polymorphism of T / C at position 125991240 on chromosome 6 of Camellia oleifera;

[0026] SNP-7-134929316, which contains a nucleotide sequence with a polymorphism of T / C at position 134929316 on chromosome 7 of Camellia oleifera;

[0027] SNP-8-37227126, which contains a nucleotide sequence with a polymorphism of C / T at position 37227126 of chromosome 8 of Camellia oleifera;

[0028] SNP-9-10583849, which contains a nucleotide sequence with a polymorphism of A / T at position 10583849 of chromosome 9 of Camellia oleifera;

[0029] SNP-9-23224047, which contains a nucleotide sequence with a polymorphism of T / C at position 23224047 on chromosome 9 of Camellia oleifera;

[0030] SNP-9-89305797, which contains a nucleotide sequence with a polymorphism of T / C at position 89305797 on chromosome 9 of Camellia oleifera;

[0031] SNP-9-95564863, which contains a nucleotide sequence with a polymorphism of G / T at position 95564863 of chromosome 9 of Camellia oleifera;

[0032] SNP-10-146191194, which contains a nucleotide sequence with a polymorphism of G / C at position 146191194 of chromosome 10 of Camellia oleifera;

[0033] SNP-11-126349400, which contains a nucleotide sequence with a polymorphism of C / T at position 126349400 on chromosome 11 of Camellia oleifera;

[0034] SNP-11-150316502, which contains a nucleotide sequence with a polymorphism of G / A at position 150316502 of chromosome 11 of Camellia oleifera;

[0035] SNP-11-175529761, which contains a nucleotide sequence with a polymorphism of G / C at position 175529761 of chromosome 11 of Camellia oleifera;

[0036] SNP-12-38432972, which contains a nucleotide sequence with a polymorphism of C / G at position 38432972 of chromosome 12 of Camellia oleifera;

[0037] SNP-12-88588135, which contains a nucleotide sequence with a polymorphism of G / A at position 88588135 of chromosome 12 of Camellia oleifera;

[0038] SNP-13-180557942, which contains a nucleotide sequence with a polymorphism of T / C at position 180557942 of chromosome 13 of Camellia oleifera;

[0039] SNP-14-86914708, which contains a nucleotide sequence with a polymorphism of T / C at position 86914708 on chromosome 14 of Camellia oleifera;

[0040] SNP-14-87149383 contains a nucleotide sequence with a polymorphism of G / T located at position 87149383 of chromosome 14 of Camellia oleifera.

[0041] The positions of the polymorphic sites of the above-mentioned SNP molecular markers are determined based on the camellia oleifera genome sequence with the version number CON genome v1.0.

[0042] Specifically, the SNP molecular markers for identifying oil-tea camellia varieties provided by the present invention include any one or more combinations of the following SNP molecular markers:

[0043] SNP-1-16341109, which contains a nucleotide sequence with a polymorphism of A / G at position 290 of the sequence shown in SEQ ID NO.1;

[0044] SNP-1-22551920, which contains a nucleotide sequence with a polymorphism of C / T at position 286 of the sequence shown in SEQ ID NO.2;

[0045] SNP-1-34038354, which contains a nucleotide sequence in which the polymorphism at position 254 is T / A as shown in SEQ ID NO.3;

[0046] SNP-1-49052505, which contains a nucleotide sequence with a polymorphism of A / G at position 269 of the sequence shown in SEQ ID NO. 4;

[0047] SNP-1-110833857, which contains a nucleotide sequence with a polymorphism of G / C at position 284 of the sequence as shown in SEQ ID NO.5;

[0048] SNP-1-115890029, which contains a nucleotide sequence with a polymorphism of A / G at position 253 of the sequence as shown in SEQ ID NO.6;

[0049] SNP-1-188555309, which contains a nucleotide sequence with a polymorphism of T / G at position 246 of the sequence as shown in SEQ ID NO.7;

[0050] SNP-2-4697104, which contains a nucleotide sequence with a polymorphism of T / A at position 247 of the sequence as shown in SEQ ID NO.8;

[0051] SNP-2-102046792, which contains a nucleotide sequence with a polymorphism of C / A at position 288 of the sequence as shown in SEQ ID NO.9;

[0052] SNP-3-58564519, which contains a nucleotide sequence in which the polymorphism at position 265 is C / T as shown in SEQ ID NO.10;

[0053] SNP-4-5589980, which contains a nucleotide sequence with a polymorphism of T / G at position 252 of the sequence as shown in SEQ ID NO.11;

[0054] SNP-5-59238652, which contains a nucleotide sequence with a polymorphism of G / C at position 187 of the sequence as shown in SEQ ID NO.12;

[0055] SNP-5-135461353, which contains a nucleotide sequence with a polymorphism of T / C at position 282 of the sequence as shown in SEQ ID NO. 13;

[0056] SNP-5-175443498, which contains a nucleotide sequence with a polymorphism of T / G at position 272 of the sequence as shown in SEQ ID NO.14;

[0057] SNP-6-123202382, which contains a nucleotide sequence with a polymorphism of A / C at position 274 of the sequence as shown in SEQ ID NO.15;

[0058] SNP-6-125991240, which contains a nucleotide sequence with a polymorphism of T / C at position 200 of the sequence as shown in SEQ ID NO.16;

[0059] SNP-7-134929316, which contains a nucleotide sequence with a polymorphism of T / C at position 233 of the sequence as shown in SEQ ID NO.17;

[0060] SNP-8-37227126, which contains a nucleotide sequence with a polymorphism of C / T at position 276 of the sequence as shown in SEQ ID NO. 18;

[0061] SNP-9-10583849, which contains a nucleotide sequence in which the polymorphism at position 259 is A / T as shown in SEQ ID NO. 19;

[0062] SNP-9-23224047, which contains a nucleotide sequence with a polymorphism of T / C at position 254 of the sequence as shown in SEQ ID NO. 20;

[0063] SNP-9-89305797, which contains a nucleotide sequence with a polymorphism of T / C at position 276 of the sequence as shown in SEQ ID NO. 21;

[0064] SNP-9-95564863, which contains a nucleotide sequence with a polymorphism of G / T at position 289 of the sequence as shown in SEQ ID NO. 22;

[0065] SNP-10-146191194, which contains a nucleotide sequence with a polymorphism of G / C at position 240 as shown in SEQ ID NO. 23;

[0066] SNP-11-126349400, which contains a nucleotide sequence with a polymorphism of C / T at position 227 of the sequence as shown in SEQ ID NO. 24;

[0067] SNP-11-150316502, which contains a nucleotide sequence with a polymorphism of G / A at position 238 of the sequence as shown in SEQ ID NO.25;

[0068] SNP-11-175529761, which contains a nucleotide sequence with a polymorphism of G / C at position 255 of the sequence as shown in SEQ ID NO.26;

[0069] SNP-12-38432972, which contains a nucleotide sequence with a polymorphism of C / G at position 227 of the sequence as shown in SEQ ID NO. 27;

[0070] SNP-12-88588135, which contains a nucleotide sequence with a polymorphism of G / A at position 253 of the sequence as shown in SEQ ID NO.28;

[0071] SNP-13-180557942, which contains a nucleotide sequence with a polymorphism of T / C at position 282 of the sequence as shown in SEQ ID NO. 29;

[0072] SNP-14-86914708, which contains a nucleotide sequence with a polymorphism of T / C at position 224 of the sequence as shown in SEQ ID NO.30;

[0073] SNP-14-87149383 contains a nucleotide sequence in which the polymorphism at position 257 is G / T as shown in SEQ ID NO.31.

[0074] Preferably, the sequences of the above-mentioned SNP molecular markers are as follows: the nucleotide sequence of SNP-1-16341109 is shown in SEQ ID NO.1, and the polymorphism at position 290 is A / G; the nucleotide sequence of SNP-1-22551920 is shown in SEQ ID NO.2, and the polymorphism at position 286 is C / T; the nucleotide sequence of SNP-1-34038354 is shown in SEQ ID NO.3, and the polymorphism at position 254 is T / A; the nucleotide sequence of SNP-1-49052505 is shown in SEQ ID NO.4, and the polymorphism at position 269 is A / G; the nucleotide sequence of SNP-1-110833857 is shown in SEQ ID NO.5, and the polymorphism at position 284 is G / C; the nucleotide sequence of SNP-1-115890029 is shown in SEQ ID NO.6. The nucleotide sequence of SNP-1-188555309 is shown in SEQ ID NO.7, and the polymorphism at position 246 is T / G; the nucleotide sequence of SNP-2-4697104 is shown in SEQ ID NO.8, and the polymorphism at position 247 is T / A; the nucleotide sequence of SNP-2-102046792 is shown in SEQ ID NO.9, and the polymorphism at position 288 is C / A; the nucleotide sequence of SNP-3-58564519 is shown in SEQ ID NO.10, and the polymorphism at position 265 is C / T; the nucleotide sequence of SNP-4-5589980 is shown in SEQ ID NO.11, and the polymorphism at position 252 is T / G; the nucleotide sequence of SNP-5-59238652 is shown in SEQ ID The nucleotide sequence of SNP-5-135461353 is shown in SEQ ID NO.13, and the polymorphism at position 282 is T / C; the nucleotide sequence of SNP-5-175443498 is shown in SEQ ID NO.14, and the polymorphism at position 272 is T / G; the nucleotide sequence of SNP-6-123202382 is shown in SEQ ID NO.15, and the polymorphism at position 274 is A / C; the nucleotide sequence of SNP-6-125991240 is shown in SEQ ID NO.16, and the polymorphism at position 200 is T / C; the nucleotide sequence of SNP-7-134929316 is shown in SEQ ID NO.17, and the polymorphism at position 233 is T / C; the nucleotide sequence of SNP-8-37227126 is shown in SEQ ID The nucleotide sequence of SNP-9-10583849 is shown in SEQ ID NO.19, and the polymorphism at position 259 is A / T; the nucleotide sequence of SNP-9-23224047 is shown in SEQ ID NO.The nucleotide sequence of SNP-9-89305797 is shown in SEQ ID NO.21, and the polymorphism at position 276 is T / C; the nucleotide sequence of SNP-9-95564863 is shown in SEQ ID NO.22, and the polymorphism at position 289 is G / T; the nucleotide sequence of SNP-10-146191194 is shown in SEQ ID NO.23, and the polymorphism at position 240 is G / C; the nucleotide sequence of SNP-11-126349400 is shown in SEQ ID NO.24, and the polymorphism at position 227 is C / T; the nucleotide sequence of SNP-11-150316502 is shown in SEQ ID NO.25, and the polymorphism at position 238 is G / A; the nucleotide sequence of SNP-11-175529761 is shown in SEQ ID NO. The nucleotide sequence of SNP-12-38432972 is shown in SEQ ID NO.27, and the polymorphism at position 227 is C / G; the nucleotide sequence of SNP-12-88588135 is shown in SEQ ID NO.28, and the polymorphism at position 253 is G / A; the nucleotide sequence of SNP-13-180557942 is shown in SEQ ID NO.29, and the polymorphism at position 282 is T / C; the nucleotide sequence of SNP-14-86914708 is shown in SEQ ID NO.30, and the polymorphism at position 224 is T / C; the nucleotide sequence of SNP-14-87149383 is shown in SEQ ID NO.31, and the polymorphism at position 257 is G / T. .

[0075] In some embodiments of the present invention, the SNP molecular marker used to identify oil-tea camellia varieties is SNP-1-110833857.

[0076] In some embodiments of the present invention, a SNP molecular marker combination for identifying oil-tea camellia varieties is provided, which includes SNP-1-34038354, SNP-1-110833857 and SNP-11-175529761.

[0077] In some embodiments of the present invention, a SNP molecular marker combination for identifying oil-tea camellia varieties is provided, which includes SNP-1-34038354, SNP-1-110833857, SNP-11-175529761, SNP-14-86914708, SNP-14-87149383, SNP-2-4697104, SNP-5-175443498 and SNP-9-23224047. The molecular marker combination contains 8 core SNP molecular markers, which can realize the identification of 221 oil-tea camellia clones described in the present invention.

[0078] In some embodiments of the present invention, a SNP molecular marker combination for identifying oil-tea camellia varieties is provided, which includes the 31 SNP molecular markers described above: SNP-1-16341109, SNP-1-22551920, SNP-1-34038354, SNP-1-49052505, SNP-1-110833857, SNP-1-115890029, SNP-1-188555309, SNP-2-4697104, SNP-2-102046792, SNP-3-58564519, SNP-4-5589980, SNP-5-59238652, SNP-5-135461353, SNP-5-175443498, SNP -6-123202382、SNP-6-125991240、SNP-7-134929316、SNP-8-37227126、SNP-9-1 0583849、SNP-9-23224047、SNP-9-89305797、SNP-9-95564863、SNP-10-1461911 94, SNP-11-126349400, SNP-11-150316502, SNP-11-175529761, SNP-12-38432972, SNP-12-88588135, SNP-13-180557942, SNP-14-86914708, SNP-14-87149383. The molecular marker combination comprises 31 SNP molecular markers, which can realize the identification of 221 oil-tea camellia clones of the present invention with high accuracy.

[0079] In a second aspect, the present invention provides a KASP primer set for detecting the SNP molecular markers described above, the primer set comprising one or more of the following primer pairs: SEQ ID NO.32-34, SEQ ID NO.35-37, SEQ ID NO.38-40, SEQ ID NO.41-43, SEQ ID NO.44-46, SEQ ID NO.47-49, SEQ ID NO.50-52, SEQ ID NO.53-55, SEQ ID NO.56-58, SEQ ID NO.59-61, SEQ ID NO.62-64, SEQ ID NO.65-67, SEQ ID NO.68-70, SEQ ID NO.71-73, SEQ ID NO.74-76, SEQ ID NO.77-79, SEQ ID NO.80-82, SEQ ID NO.83-85, SEQ ID NO.86-88, SEQ ID NO.89-91, SEQ ID NO.92-94, SEQ ID NO.95-96, SEQ ID NO.97-98, SEQ ID NO.99-100, SEQ ID NO.101-102, SEQ ID NO.103-104, SEQ ID NO.105-106, SEQ ID NO.107-108 NO.95-97, SEQ ID NO.98-100, SEQ ID NO.101-103, SEQ ID NO.104-106, SEQ ID NO.107-109, SEQ ID NO.110-112, SEQ ID NO.113-115, SEQ ID NO.116-118, SEQ ID NO.119-121, SEQ IDNO.122-124.

[0080] Specifically, the nucleotide sequences of the primers described above are shown in Table 1.

[0081] Table 1 Primer sequences

[0082]

[0083] In a third aspect, the present invention provides a kit comprising the above primer set.

[0084] In addition to the primer set, the above kit may also include other reagents for SNP genotyping, such as PCR reaction reagents (including but not limited to DNA polymerase, reaction buffer, dNTP) and the like.

[0085] In a fourth aspect, the present invention provides a DNA chip comprising the above primer set.

[0086] In a fifth aspect, the present invention provides any of the following applications of the above-mentioned SNP molecular marker, the primer set, the kit or the DNA chip:

[0087] (1) Application in identifying Camellia oleifera variety resources;

[0088] (2) Application in constructing DNA fingerprints of Camellia oleifera varieties;

[0089] (3) Application in the construction of molecular identity cards for Camellia oleifera varieties.

[0090] (4) Application in authenticity or purity testing of tea oil seeds or seedlings;

[0091] (5) Application in the traceability of oil-tea camellia varieties;

[0092] (6) Application in molecular marker-assisted breeding of Camellia oleifera.

[0093] In the above application, the oil-tea camellia is common oil-tea camellia.

[0094] In a sixth aspect, the present invention provides a method for constructing a molecular identity card of a tea oil variety, the method comprising: performing genotyping on the SNP molecular markers mentioned above in the genome of the tea oil sample, and constructing a molecular identity card of the tea oil variety based on the genotyping results in combination with the sample origin, resource type and subpopulation distribution.

[0095] Preferably, the genotyping method comprises: using the genomic DNA of the oil-tea camellia sample as a template, performing PCR amplification using the primer set described above to obtain a PCR amplification product, performing fluorescence detection on the PCR amplification product, and determining the genotyping of the SNP molecular marker according to the fluorescence type.

[0096] In a seventh aspect, the present invention provides a method for identifying oil-tea camellia varieties, the method comprising: performing genotyping on the SNP molecular markers mentioned above in the genome of the oil-tea camellia sample to be identified, and judging the variety of the oil-tea camellia sample based on the genotyping results in combination with the sample origin, resource type and subpopulation distribution.

[0097] Preferably, the genotyping method comprises: using the genomic DNA of the oil-tea camellia sample as a template, performing PCR amplification using the primer set described above to obtain a PCR amplification product, performing fluorescence detection on the PCR amplification product, and determining the genotyping of the SNP molecular marker according to the fluorescence type.

[0098] In the present invention, the oil-tea camellia is common oil-tea camellia, preferably one or more of the 221 oil-tea camellia varieties shown in Table 2, Table 3 and Table 4.

[0099] The beneficial effects of the present invention are as follows: the 31 core SNP molecular markers for the identification of oil-tea camellia variety resources provided by the present invention can accurately distinguish all tested oil-tea camellia variety resources, and provide effective molecular tools and methods for the identification of oil-tea camellia varieties, variety tracing, and authenticity and purity detection of seedlings. The DNA fingerprint of oil-tea camellia variety resources constructed by the 31 SNP molecular markers provided by the present invention is combined with the basic attribute information such as the origin, resource type and subpopulation distribution of the variety resources to construct a unique molecular identity card for each oil-tea camellia variety resource, and generate corresponding barcodes and QR codes, providing technical support for the rapid detection of oil-tea camellia variety quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0100] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0101] Figure 1 The statistical characteristics of SNPs in the genome of the Camellia oleifera population in Example 1 of the present invention include: A. SNP variation type statistics; B. SNP distribution position statistics on the genome; C. SNP function statistics located in the gene coding region.

[0102] Figure 2 The results of Sanger sequencing of some core SNP sites in Example 1 of the present invention are as follows

[0103] Figure 3 These are the genotype results of KASPar analysis of some core SNPs sites in Example 3 of the present invention. DETAILED DESCRIPTION

[0104] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0105] The 221 common camellia oleifera clones (varieties) materials used in the following examples originated from major camellia oleifera producing areas in China's subtropical regions and are all preserved in the camellia oleifera germplasm resource bank of the National Camellia and Tung Oil Seed Base located in the Dongfanghong Forest Farm, Wucheng District, Jinhua City, Zhejiang Province. The basic information of each material is shown in Tables 2, 3 and 4.

[0106] Table 2 Information on 221 common Camellia oleifera clones (1)

[0107]

[0108] Table 3 Information on 221 common Camellia oleifera clones (2)

[0109]

[0110] Table 4 Information on 221 common Camellia oleifera clones (3)

[0111]

[0112] Note: In Tables 2, 3, and 4, , 33 for Zhejiang, 35 for Fujian, 36 for Jiangxi, 43 for Hunan, 44 for Guangdong, 45 for Guangxi, XX means the origin is unknown;

[0113] #, A: wild resources (clones), B: bred varieties;

[0114] &, the sample clustering information is defined according to the genetic structure of the population, and the specific reference is Lin et al. (Lin P, Wang K, Wang Y, et al . The genome of oil-Camellia and population genomics analysis provide insights into seed oil domestication. Genome Biology, 2022, 23:14.).

[0115] Example 1 RNA extraction, transcriptome sequencing and SNP identification of Camellia oleifera materials

[0116] 1. RNA extraction

[0117] RNA was extracted from 221 common Camellia oleifera clones (varieties) as follows: In August 2017, developing kernels were collected and quickly frozen with liquid nitrogen and stored in a -80°C ultra-low temperature refrigerator. Total RNA was extracted using an RNA extraction kit suitable for polysaccharide- and polyphenol-rich plants (RNAprep Pure Plus Kit, Tiangen). RNA integrity and purity were tested using 1.2% agarose gel and NanoPhotometer spectrophotometer (Quawell Technology, Inc., USA), RNA concentration was accurately quantified using Qubit2.0 Fluorometer, and RNA integrity was further accurately tested using Agilent 2100 bioanalyzer.

[0118] 2. Transcriptome sequencing

[0119] After the total RNA samples of each sample were tested and qualified, the RNAseq library of Camellia oleifera kernel was constructed. The RNAseq library was sequenced bidirectionally at 150 bp on the Illumina HiSeqTM 4000 platform, and the sequencing volume of each library was 6G bp. The transcriptome sequencing data can be viewed at NCBI (SRR14934120~SRR14934340 under Bioproject accession No. PRJNA732216).

[0120] 3. Sequencing data processing and SNP identification

[0121] Methods: Sequencing data filtering and alignment, SNP identification and filtering were performed according to the literature (Lin et al. 2022). GATK4 was used to perform statistics on the function, location and mutation type of the variant sites.

[0122] Results: A total of 1,849,953 SNPs were detected in the Camellia oleifera population. The main variation types of these SNPs were base conversions, accounting for 69.65% of all SNPs, and base transversions accounted for 30.35% ( Figure 1 A), the transition transversion ratio (Ti / Tv) is 2.29. These SNPs are mostly located in the intergenic region, followed by the exon region and the intron region ( Figure 1 There are 363,812 SNPs located in the coding region of genes, which can be divided into four categories according to their functions: synonymous mutations, non-synonymous mutations, nonsense mutations and stop codon loss. Synonymous mutations and non-synonymous mutations are the main types of SNPs located in the coding region, accounting for 99.41% ( Figure 1 C).

[0123] 4. Screening and verification of core SNP loci for identification of oil-tea camellia varieties

[0124] Methods: SNPs markers in the entire transcriptome of all samples were filtered three times to obtain the core loci for the identification of oil-tea camellia varieties. The first filtering criteria were: (1) the typing missing rate in all samples was <20%; (2) the minimum allele frequency MAF Reaching above 0.1; (3) The site conforms to the Hard-Wenberg equilibrium; (4) The polymorphic information content PIC (5) the linkage disequilibrium decay distance LD is above 0.3; (6) there are no other SNP sites within 100 bp before and after the site; (7) the site is located in the exon of the gene; (8) there are only two alleles at the variant site; (9) the number of reads supported by each site in the sequencing results of each sample is more than 10; (10) the SNP sites are distributed as evenly as possible on all chromosomes, and the sites on chromosomes with too many sites are retained at a density of 100 kb / SNP. The second round of marker filtering uses a combination of PCA and random forest analysis. PCA screening markers are performed using pcadapt v4.3.2 (https: / / bcm-uga.github.io / pcadapt / articles / pcadapt.html), and random forest analysis is implemented using the R software package "Random forest". The SNP markers obtained after the first two rounds of screening are further filtered in the third round, and the filtering standard is that the sample typing missing rate is <10%. The core SNP sites are obtained and their genotypes in all samples are clarified.

[0125] 4 μL RNA was used as template for each sample, and one-strand cDNA was synthesized according to the First-Strand Synthesis System kit manual of Invitrogen, USA. Primers were designed within the range of about 200 bp upstream and downstream of the screened core SNP sites with reference to the genome sequence, and PCR amplification was performed using cDNA as a template and a high-fidelity amplification enzyme. The 50 μL PCR reaction system contained 60 ng cDNA, 2 × LAmp Master Mix (Vazyme, Nanjing, China) 25 μL, 0.4 mM forward primer and 0.4 mM reverse primer. The PCR amplification program was set to 94 °C pre-denaturation for 5 min; 35 cycles including 94 °C denaturation for 30 s, annealing at a specific annealing temperature for 30 s, 72 °C extension for 2 min, and finally 72 °C extension for 7 min, and stored at 4 °C. The amplified products were purified and recovered using 1.2% agarose, and the amplification primers were used as sequencing primers and sequenced using ABI 3730XL DNA Analyzer to verify the genotype of the key sites of each sample.

[0126] Results: The results included the typing ratio, MAF , PIC , distribution position, number of alleles and other strict screening criteria. After the first round of filtering, 165 SNP sites were obtained. After the second round of PCA and random forest joint analysis, 58 SNP sites were obtained. After the third round of screening, 31 core SNP sites with high polymorphism, low missingness and good repeatability were finally obtained. All 31 core SNP sites are moderately polymorphic. PIC The SNPs ranged from 0.4586 to 0.4999 and were distributed on the remaining 14 chromosomes of the Camellia oleifera genome except chromosome 15 (Table 5). Each locus has two alleles and is located in the exon region of the coding gene. Three of the 31 loci were located in the 3'UTR, four were located in the 5'UTR, and the remaining 24 loci were located in the coding region of the gene, forming 15 synonymous mutations and 9 non-synonymous mutations (Table 5). The genotype information of 221 Camellia oleifera clones at the 31 SNP loci is shown in Table 6.

[0127] Table 5 31 core SNP loci and their detailed information

[0128]

[0129] Table 6 Genotype information of 221 Camellia oleifera clones at 31 SNP loci

[0130]

[0131] Note: 1-31 in the first row of Table 6 represent molecular markers SNP-1-16341109, SNP-1-22551920, SNP-1-34038354, SNP-1-49052505, SNP-1-110833857, SNP-1-115890029, SNP-1-188555309, SNP-2-4697104, SNP-2-102046792, SNP-3-58564519, SNP-4-5589980, SNP-5-59238652, SNP-5-135461353, SNP-5-175443498, SNP-6-123202382, SNP-7 P-6-125991240, SNP-7-134929316, SNP-8-37227126, SNP-9-10583849, SNP -9-23224047、SNP-9-89305797、SNP-9-95564863、SNP-10-146191194、SNP-1 1-126349400, SNP-11-150316502, SNP-11-175529761, SNP-12-38432972, S NP-12-88588135, SNP-13-180557942, SNP-14-86914708 and SNP-14-87149383.

[0132] High-fidelity enzymes were further used to perform PCR amplification on 8 of the 31 core sites (Table 7), and the amplified products were sequenced by Sanger to verify the genotypes of all samples at these 8 sites. The primers used for PCR amplification and sequencing of amplified products are shown in Table 7. A total of 1755 SNP data were successfully detected, of which 1662 sites were consistent with the transcriptome sequencing results (Table 8), with a verification rate of 94.70%. Except for the SNP-11-175529761 site, which had a slightly lower verification rate (91.36%), the verification rates of the remaining sites were basically the same. A statistical analysis was performed on the verification number of different genotypes at each site (Table 8), and it was found that the verification rates of different genotypes were slightly different. For example, the verification rate of individuals with the TT genotype at the SNP-14-86914708 site was significantly lower than that of the GG genotype ( Figure 2 ).

[0133] In summary, the transcriptome sequencing and Sanger sequencing results of these 8 SNP sites are highly consistent, which shows that the genotype data of the 31 SNP sites screened by the present invention are reliable and rich in polymorphism. It is feasible to use them for the subsequent construction of molecular fingerprints and molecular ID cards of various varieties.

[0134] Table 7 Primer information for Sanger sequencing of 8 core sites

[0135]

[0136] Table 8 Sanger sequencing verification results of 8 core SNP sites

[0137]

[0138] Example 2 Construction of Molecular Fingerprints and Molecular ID Cards of Camellia oleifera Varieties

[0139] For the genotypes of the 31 core SNP sites verified in Example 1, A, T, C, and G were converted to 1, 2, 3, and 4 in sequence, and the missing sites were replaced with 0 to construct a molecular fingerprint map specific to each variety. The germplasm resource information and the molecular fingerprint map were combined to construct the molecular ID cards of 221 Camellia oleifera clones using a barcode generator and a QR code generator (http: / / qr-batch.com / ).

[0140] According to the genotypes of 31 core SNP sites in 221 Camellia oleifera clones, the bases A, T, C, and G were converted to 1, 2, 3, and 4 in sequence, and the missing sites were assigned a value of 0. The molecular fingerprint map specific to each clone was constructed by subgroup. Among them, the single marker SNP-1-110833857 can identify 'Changlin 228' in subgroup 2 and 'Changlin 102' in subgroup 8. The three-marker combination of SNP-1-34038354, SNP-1-110833857 and SNP-11-175529761 can identify (1) Changlin 172, 561, 55, 60, 151, 252 and Gan 4 (subgroup I); (2) Changlin 54, 11, 540 and Tong 10 (subgroup II); (3) Changlin 531, 3007, 160 and fy7 (subgroup III); (4) Changlin 222, 21, 168, 64, 155, 95, 4, 77, 89, Gui 2 and Xiang 7 (subgroup IV); (5) Changlin 28, Xiang 47, Xiang 8 , Minyou 48, Xiang 4, Yajia 2, Yawu 7 and Minyou 11 (subgroup V); (6) Changlin 63, 67, Tong 29, Tong 30, Tong 40, Wuyiwu 2 and Gan 1 (subgroup VI); (7) Changlin 100, Guiruan 3, Xiang 9, Yajia 1 and Guiruan 23 (subgroup VII); (8) Changlin 8, 61 (subgroup VIII) and nearly 50 other asexual lines.

[0141] The genotypes of the 31 core loci in each clone were further converted into a 62-digit fingerprint code, and together with the germplasm information code, a variety-specific 66-digit molecular ID code was constructed (the first 4 digits are the germplasm information code, and the last 62 digits are the 31 core loci SNP-1-16341109, SNP-1-22551920, SNP-1-34038354, SNP-1-490525 05. SNP-1-110833857, SNP-1-115890029, SNP-1-188555309, SNP-2-4697104, SNP-2-1 02046792, SNP-3-58564519, SNP-4-5589980, SNP-5-59238652, SNP-5-135461353, SNP- 5-175443498、SNP-6-123202382、SNP-6-125991240、SNP-7-134929316、SNP-8-372271 26. SNP-9-10583849, SNP-9-23224047, SNP-9-89305797, SNP-9-95564863, SNP-10-14 6191194、SNP-11-126349400、SNP-11-150316502、SNP-11-175529761、SNP-12-384329 72. Genotypes of SNP-12-88588135, SNP-13-180557942, SNP-14-86914708 and SNP-14-87149383). The germplasm information code has a total of 4 digits, divided into 3 parts. The first and second digits represent the province of origin of the asexual line, such as 33 for Zhejiang, 35 for Fujian, 36 for Jiangxi, 43 for Hunan, 44 for Guangdong, 45 for Guangxi, etc. If the province of origin is unknown, use XX instead; the third digit is the germplasm resource category, referring to the "Technical Specifications for Survey and Cataloging of Oil-Palm Genetic Resources" (LY / T 2247-2014), 1 represents wild resources (populations), 2 represents wild resources (families), 3 represents wild resources (asexual lines), 4 represents local varieties, 5 represents selected varieties, 6 represents selected lines, and 7 represents genetic data; the fourth digit is the subpopulation number, assigned values ​​1 to 8.Taking 'Changlin 53' as an example, its molecular ID number is 335444322144431122211132224423243323232212230022443244433341233342, indicating that the origin is Zhejiang Province, it is a selected variety, belongs to subgroup 4, and has 31 core markers (SNP-1-16341109, SNP-1-22551920, SNP-1-34038354, SNP-1-4905250 5. SNP-1-110833857, SNP-1-115890029, SNP-1-188555309, SNP-2-4697104, SNP-2-102046792, SNP-3-58 564519, SNP-4-5589980, SNP-5-59238652, SNP-5-135461353, SNP-5-175443498, SNP-6-123202382, SNP- 6-125991240、SNP-7-134929316、SNP-8-37227126、SNP-9-10583849、SNP-9-23224047、SNP-9-89305797、 SNP-9-95564863, SNP-10-146191194, SNP-11-126349400, SNP-11-150316502, SNP-11-175529761, SNP-1 2-38432972, SNP-12-88588135, SNP-13-180557942, SNP-14-86914708, and SNP-14-87149383) were 44, 32, 21, 44, 43, 11, 22, 21, 11, 32, 22, 44, 23, 24, 33, 23, 23, 22, 12, 23, 00, 22, 44, 32, 44, 43, 33, 41, 23, 33, and 42. Finally, the 66-bit molecular ID number of each clone was used to make variety-specific barcodes and QR codes.

[0142] Example 3 Detection method of core SNP locus genotype for variety identification

[0143] Based on the competitive allele-specific PCR (kompetitive allele-specific PCR, KASPar) platform, with the genomic DNA of Camellia oleifera as the template, primers were developed for the 31 core SNP sites screened in Example 1 for rapid detection of the genotype of the variety to be identified. Three primers were designed for each core SNP marker, including two allele-specific upstream (or downstream) primers and a shared downstream (or upstream) primer, of which 14 sites included two upstream primers for distinguishing different genotypes of the SNP site and a shared downstream primer, and 17 sites had two downstream primers for distinguishing different genotypes of the SNP site and a shared upstream primer. The primer sequences are shown in Table 1.

[0144] 221 samples from the Camellia oleifera population (shown in Table 2-4) were used as test samples. KASP primers were used to perform PCR amplification of the core loci. The core loci genotypes of the test samples were analyzed by detecting the fluorescence signal to verify the genotyping effect and accuracy of the core loci KASP primers and detection method. The KASPar reaction system, PCR procedure, and fluorescence signal detection were performed according to Su et al. (Su T, Li P, Yang J, et al . Development of cost-effective single nucleotide polymorphism marker assays for genetic diversity analysis in Brassica rapa. Molecular Breeding, 2018, 38:42.) method was used, and data analysis was completed using Bio-Rad CFX Maestro software.

[0145] After comparison, the consistency rate between the genotyping results of the samples by KASP primers and the high-throughput sequencing results reached more than 90% ( Figure 3 ), which can be used for rapid genotyping in subsequent identification of Camellia oleifera varieties.

[0146] In summary, in order to construct a molecular identification system for common tea varieties, the present invention comprehensively detected SNP sites in the transcriptome of common tea; PIC, LD and other indicators were screened, and SNP-1-16341109, SNP-1-22551920, SNP-1-34038354, SNP-1-49052505, SNP-1-110833857, SNP-1-115890029, SNP-1-188555309, SNP-2-4697104, SNP-2-102046792, SNP-3 -58564519、SNP-4-5589980、SNP-5-59238652、SNP-5-135461353、SNP-5-175443498、SNP-6- 123202382、SNP-6-125991240、SNP-7-134929316、SNP-8-37227126、SNP-9-10583849、SNP-9- 23224047, SNP-9-89305797, SNP-9-95564863, SNP-10-146191194, SNP-11-126349400, SNP- 11-150316502、SNP-11-175529761、SNP-12-38432972、SNP-12-88588135、SNP-13-180557942 , SNP-14-86914708 and SNP-14-87149383, which are 31 core SNP loci, can effectively distinguish 221 Camellia oleifera clones. Combined with the origin, resource type and subpopulation distribution of the samples, the molecular fingerprint and molecular identity card of the 221 participating asexual lines were constructed, which provided a technical solution for the identification and traceability of Camellia oleifera varieties, as well as the authenticity and purity testing of seedlings, and guaranteed the quality of seedlings for the healthy development of the Camellia oleifera industry.

[0147] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A KASP primer set for detecting SNP molecular markers, characterized in that: The primer set includes the following primer pairs: SEQ ID NO.32-34, SEQ ID NO.35-37, SEQ ID NO.38-40, SEQ ID NO.41-43, SEQ ID NO.44-46, SEQ ID NO.47-49, SEQ ID NO.50-52, SEQ ID NO.53-55, SEQ ID NO.56-58, SEQ ID NO.59-61, SEQ ID NO.62-64, SEQ ID NO.65-67, SEQ ID NO.68-70, SEQ ID NO.71-73, SEQ ID NO.74-76, SEQ ID NO.77-79, SEQ ID NO.80-82, SEQ ID NO.83-85, SEQ ID NO.86-88, SEQ ID NO.89-91, SEQ ID NO.92-94, SEQ ID NO.95-97, SEQ ID NO.98-100, SEQ ID NO. NO.101-103, SEQ ID NO.104-106, SEQ ID NO.107-109, SEQ ID NO.110-112, SEQ ID NO.113-115, SEQ ID NO.116-118, SEQ ID NO.119-121, and SEQ ID NO.122-124; The SNP molecular markers include a combination of the following SNP molecular markers: SNP-1-16341109, which is the polymorphic base at position 290 of the sequence shown in SEQ ID NO.1, and its base polymorphism is A / G; SNP-1-22551920, which is the polymorphic base at position 286 of the sequence shown in SEQ ID NO. 2, and its base polymorphism is C / T; SNP-1-34038354, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 3, and its base polymorphism is T / A; SNP-1-49052505, which is the polymorphic base at position 269 of the sequence shown in SEQ ID NO. 4, and its base polymorphism is A / G; SNP-1-110833857, which is the polymorphic base at position 284 of the sequence shown in SEQ ID NO.5, and its base polymorphism is G / C; SNP-1-115890029, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO.6, and its base polymorphism is A / G; SNP-1-188555309, which is the polymorphic base at position 246 of the sequence shown in SEQ ID NO.7, and its base polymorphism is T / G; SNP-2-4697104, which is the polymorphic base at position 247 of the sequence shown in SEQ ID NO. 8, and its base polymorphism is T / A; SNP-2-102046792, which is the polymorphic base at position 288 of the sequence shown in SEQ ID NO.9, and its base polymorphism is C / A; SNP-3-58564519, which is the polymorphic base at position 265 of the sequence shown in SEQ ID NO. 10, and its base polymorphism is C / T; SNP-4-5589980, which is the polymorphic base at position 252 of the sequence shown in SEQ ID NO. 11, and its base polymorphism is T / G; SNP-5-59238652, which is the polymorphic base at position 187 of the sequence shown in SEQ ID NO. 12, and its base polymorphism is G / C; SNP-5-135461353, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 13, and its base polymorphism is T / C; SNP-5-175443498, which is the polymorphic base at position 272 of the sequence shown in SEQ ID NO. 14, and its base polymorphism is T / G; SNP-6-123202382, which is the polymorphic base at position 274 of the sequence shown in SEQ ID NO. 15, and its base polymorphism is A / C; SNP-6-125991240, which is the polymorphic base at position 200 of the sequence shown in SEQ ID NO. 16, and its base polymorphism is T / C; SNP-7-134929316, which is the polymorphic base at position 233 of the sequence shown in SEQ ID NO. 17, and its base polymorphism is T / C; SNP-8-37227126, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 18, and its base polymorphism is C / T; SNP-9-10583849, which is the polymorphic base at position 259 of the sequence shown in SEQ ID NO. 19, and its base polymorphism is A / T; SNP-9-23224047, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 20, and its base polymorphism is T / C; SNP-9-89305797, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 21, and its base polymorphism is T / C; SNP-9-95564863, which is the polymorphic base at position 289 of the sequence shown in SEQ ID NO. 22, and its base polymorphism is G / T; SNP-10-146191194, which is the polymorphic base at position 240 of the sequence shown in SEQ ID NO. 23, and its base polymorphism is G / C; SNP-11-126349400, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 24, and its base polymorphism is C / T; SNP-11-150316502, which is the polymorphic base at position 238 of the sequence shown in SEQ ID NO. 25, and its base polymorphism is G / A; SNP-11-175529761, which is the polymorphic base at position 255 of the sequence shown in SEQ ID NO. 26, and its base polymorphism is G / C; SNP-12-38432972, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 27, and its base polymorphism is C / G; SNP-12-88588135, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO. 28, and its base polymorphism is G / A; SNP-13-180557942, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 29, and its base polymorphism is T / C; SNP-14-86914708, which is the polymorphic base at position 224 of the sequence shown in SEQ ID NO. 30, and its base polymorphism is T / C; SNP-14-87149383 is the polymorphic base at position 257 of the sequence shown in SEQ ID NO. 31, and its base polymorphism is G / T.

2. A kit, characterized in that: The kit comprises the primer set according to claim 1.

3. A DNA chip, characterized in that The DNA chip comprises the primer set according to claim 1.

4. Any of the following applications of reagents for detecting SNP molecular markers: (1) Application in identifying Camellia oleifera variety resources; (2) Application in constructing DNA fingerprints of Camellia oleifera varieties; (3) Application in the construction of molecular ID cards for Camellia oleifera varieties; (4) Application in authenticity or purity testing of tea oil seeds or seedlings; (5) Application in the traceability of oil-tea camellia varieties; (6) Application in molecular marker-assisted breeding of Camellia oleifera; in, The SNP molecular markers include a combination of the following SNP molecular markers: SNP-1-16341109, which is the polymorphic base at position 290 of the sequence shown in SEQ ID NO.1, and its base polymorphism is A / G; SNP-1-22551920, which is the polymorphic base at position 286 of the sequence shown in SEQ ID NO. 2, and its base polymorphism is C / T; SNP-1-34038354, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 3, and its base polymorphism is T / A; SNP-1-49052505, which is the polymorphic base at position 269 of the sequence shown in SEQ ID NO. 4, and its base polymorphism is A / G; SNP-1-110833857, which is the polymorphic base at position 284 of the sequence shown in SEQ ID NO.5, and its base polymorphism is G / C; SNP-1-115890029, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO.6, and its base polymorphism is A / G; SNP-1-188555309, which is the polymorphic base at position 246 of the sequence shown in SEQ ID NO.7, and its base polymorphism is T / G; SNP-2-4697104, which is the polymorphic base at position 247 of the sequence shown in SEQ ID NO. 8, and its base polymorphism is T / A; SNP-2-102046792, which is the polymorphic base at position 288 of the sequence shown in SEQ ID NO.9, and its base polymorphism is C / A; SNP-3-58564519, which is the polymorphic base at position 265 of the sequence shown in SEQ ID NO. 10, and its base polymorphism is C / T; SNP-4-5589980, which is the polymorphic base at position 252 of the sequence shown in SEQ ID NO. 11, and its base polymorphism is T / G; SNP-5-59238652, which is the polymorphic base at position 187 of the sequence shown in SEQ ID NO. 12, and its base polymorphism is G / C; SNP-5-135461353, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 13, and its base polymorphism is T / C; SNP-5-175443498, which is the polymorphic base at position 272 of the sequence shown in SEQ ID NO. 14, and its base polymorphism is T / G; SNP-6-123202382, which is the polymorphic base at position 274 of the sequence shown in SEQ ID NO. 15, and its base polymorphism is A / C; SNP-6-125991240, which is the polymorphic base at position 200 of the sequence shown in SEQ ID NO. 16, and its base polymorphism is T / C; SNP-7-134929316, which is the polymorphic base at position 233 of the sequence shown in SEQ ID NO. 17, and its base polymorphism is T / C; SNP-8-37227126, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 18, and its base polymorphism is C / T; SNP-9-10583849, which is the polymorphic base at position 259 of the sequence shown in SEQ ID NO. 19, and its base polymorphism is A / T; SNP-9-23224047, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 20, and its base polymorphism is T / C; SNP-9-89305797, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 21, and its base polymorphism is T / C; SNP-9-95564863, which is the polymorphic base at position 289 of the sequence shown in SEQ ID NO. 22, and its base polymorphism is G / T; SNP-10-146191194, which is the polymorphic base at position 240 of the sequence shown in SEQ ID NO. 23, and its base polymorphism is G / C; SNP-11-126349400, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 24, and its base polymorphism is C / T; SNP-11-150316502, which is the polymorphic base at position 238 of the sequence shown in SEQ ID NO. 25, and its base polymorphism is G / A; SNP-11-175529761, which is the polymorphic base at position 255 of the sequence shown in SEQ ID NO. 26, and its base polymorphism is G / C; SNP-12-38432972, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 27, and its base polymorphism is C / G; SNP-12-88588135, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO. 28, and its base polymorphism is G / A; SNP-13-180557942, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 29, and its base polymorphism is T / C; SNP-14-86914708, which is the polymorphic base at position 224 of the sequence shown in SEQ ID NO. 30, and its base polymorphism is T / C; SNP-14-87149383 is the polymorphic base at position 257 of the sequence shown in SEQ ID NO. 31, and its base polymorphism is G / T.

5. Any of the following uses of the primer set according to claim 1, the kit according to claim 2, or the DNA chip according to claim 3: (1) Application in identifying Camellia oleifera variety resources; (2) Application in constructing DNA fingerprints of Camellia oleifera varieties; (3) Application in the construction of molecular ID cards for Camellia oleifera varieties; (4) Application in authenticity or purity testing of tea oil seeds or seedlings; (5) Application in the traceability of oil-tea camellia varieties; (6) Application in molecular marker-assisted breeding of Camellia oleifera.

6. A method for constructing a molecular identity card of a Camellia oleifera variety, characterized in that: The method comprises: genotyping the SNP molecular markers in the genome of the oil-tea camellia sample, and constructing a molecular identity card of the oil-tea camellia variety based on the genotyping result and combining the sample origin, resource type and subpopulation distribution; Wherein, the SNP molecular markers include a combination of the following SNP molecular markers: SNP-1-16341109, which is the polymorphic base at position 290 of the sequence shown in SEQ ID NO.1, and its base polymorphism is A / G; SNP-1-22551920, which is the polymorphic base at position 286 of the sequence shown in SEQ ID NO. 2, and its base polymorphism is C / T; SNP-1-34038354, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 3, and its base polymorphism is T / A; SNP-1-49052505, which is the polymorphic base at position 269 of the sequence shown in SEQ ID NO. 4, and its base polymorphism is A / G; SNP-1-110833857, which is the polymorphic base at position 284 of the sequence shown in SEQ ID NO.5, and its base polymorphism is G / C; SNP-1-115890029, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO.6, and its base polymorphism is A / G; SNP-1-188555309, which is the polymorphic base at position 246 of the sequence shown in SEQ ID NO.7, and its base polymorphism is T / G; SNP-2-4697104, which is the polymorphic base at position 247 of the sequence shown in SEQ ID NO. 8, and its base polymorphism is T / A; SNP-2-102046792, which is the polymorphic base at position 288 of the sequence shown in SEQ ID NO.9, and its base polymorphism is C / A; SNP-3-58564519, which is the polymorphic base at position 265 of the sequence shown in SEQ ID NO. 10, and its base polymorphism is C / T; SNP-4-5589980, which is the polymorphic base at position 252 of the sequence shown in SEQ ID NO. 11, and its base polymorphism is T / G; SNP-5-59238652, which is the polymorphic base at position 187 of the sequence shown in SEQ ID NO. 12, and its base polymorphism is G / C; SNP-5-135461353, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 13, and its base polymorphism is T / C; SNP-5-175443498, which is the polymorphic base at position 272 of the sequence shown in SEQ ID NO. 14, and its base polymorphism is T / G; SNP-6-123202382, which is the polymorphic base at position 274 of the sequence shown in SEQ ID NO. 15, and its base polymorphism is A / C; SNP-6-125991240, which is the polymorphic base at position 200 of the sequence shown in SEQ ID NO. 16, and its base polymorphism is T / C; SNP-7-134929316, which is the polymorphic base at position 233 of the sequence shown in SEQ ID NO. 17, and its base polymorphism is T / C; SNP-8-37227126, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 18, and its base polymorphism is C / T; SNP-9-10583849, which is the polymorphic base at position 259 of the sequence shown in SEQ ID NO. 19, and its base polymorphism is A / T; SNP-9-23224047, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 20, and its base polymorphism is T / C; SNP-9-89305797, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 21, and its base polymorphism is T / C; SNP-9-95564863, which is the polymorphic base at position 289 of the sequence shown in SEQ ID NO. 22, and its base polymorphism is G / T; SNP-10-146191194, which is the polymorphic base at position 240 of the sequence shown in SEQ ID NO. 23, and its base polymorphism is G / C; SNP-11-126349400, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 24, and its base polymorphism is C / T; SNP-11-150316502, which is the polymorphic base at position 238 of the sequence shown in SEQ ID NO. 25, and its base polymorphism is G / A; SNP-11-175529761, which is the polymorphic base at position 255 of the sequence shown in SEQ ID NO. 26, and its base polymorphism is G / C; SNP-12-38432972, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 27, and its base polymorphism is C / G; SNP-12-88588135, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO. 28, and its base polymorphism is G / A; SNP-13-180557942, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 29, and its base polymorphism is T / C; SNP-14-86914708, which is the polymorphic base at position 224 of the sequence shown in SEQ ID NO. 30, and its base polymorphism is T / C; SNP-14-87149383 is the polymorphic base at position 257 of the sequence shown in SEQ ID NO. 31, and its base polymorphism is G / T.

7. The method according to claim 6, characterized in that The genotyping method comprises: using the genomic DNA of the oil-tea camellia sample as a template, performing PCR amplification using the primer set of claim 1 to obtain a PCR amplification product, performing fluorescence detection on the PCR amplification product, and determining the genotyping of the SNP molecular marker according to the fluorescence type.

8. A method for identifying oil-tea camellia varieties, characterized in that: The method comprises: performing genotyping on the SNP molecular markers in the genome of the oil-tea camellia sample to be identified, and judging the variety of the oil-tea camellia sample according to the genotyping result, in combination with the origin, resource type and subpopulation distribution of the sample; The SNP molecular markers include a combination of the following SNP molecular markers: SNP-1-16341109, which is the polymorphic base at position 290 of the sequence shown in SEQ ID NO.1, and its base polymorphism is A / G; SNP-1-22551920, which is the polymorphic base at position 286 of the sequence shown in SEQ ID NO. 2, and its base polymorphism is C / T; SNP-1-34038354, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 3, and its base polymorphism is T / A; SNP-1-49052505, which is the polymorphic base at position 269 of the sequence shown in SEQ ID NO. 4, and its base polymorphism is A / G; SNP-1-110833857, which is the polymorphic base at position 284 of the sequence shown in SEQ ID NO.5, and its base polymorphism is G / C; SNP-1-115890029, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO.6, and its base polymorphism is A / G; SNP-1-188555309, which is the polymorphic base at position 246 of the sequence shown in SEQ ID NO.7, and its base polymorphism is T / G; SNP-2-4697104, which is the polymorphic base at position 247 of the sequence shown in SEQ ID NO. 8, and its base polymorphism is T / A; SNP-2-102046792, which is the polymorphic base at position 288 of the sequence shown in SEQ ID NO.9, and its base polymorphism is C / A; SNP-3-58564519, which is the polymorphic base at position 265 of the sequence shown in SEQ ID NO. 10, and its base polymorphism is C / T; SNP-4-5589980, which is the polymorphic base at position 252 of the sequence shown in SEQ ID NO. 11, and its base polymorphism is T / G; SNP-5-59238652, which is the polymorphic base at position 187 of the sequence shown in SEQ ID NO. 12, and its base polymorphism is G / C; SNP-5-135461353, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 13, and its base polymorphism is T / C; SNP-5-175443498, which is the polymorphic base at position 272 of the sequence shown in SEQ ID NO. 14, and its base polymorphism is T / G; SNP-6-123202382, which is the polymorphic base at position 274 of the sequence shown in SEQ ID NO. 15, and its base polymorphism is A / C; SNP-6-125991240, which is the polymorphic base at position 200 of the sequence shown in SEQ ID NO. 16, and its base polymorphism is T / C; SNP-7-134929316, which is the polymorphic base at position 233 of the sequence shown in SEQ ID NO. 17, and its base polymorphism is T / C; SNP-8-37227126, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 18, and its base polymorphism is C / T; SNP-9-10583849, which is the polymorphic base at position 259 of the sequence shown in SEQ ID NO. 19, and its base polymorphism is A / T; SNP-9-23224047, which is the polymorphic base at position 254 of the sequence shown in SEQ ID NO. 20, and its base polymorphism is T / C; SNP-9-89305797, which is the polymorphic base at position 276 of the sequence shown in SEQ ID NO. 21, and its base polymorphism is T / C; SNP-9-95564863, which is the polymorphic base at position 289 of the sequence shown in SEQ ID NO. 22, and its base polymorphism is G / T; SNP-10-146191194, which is the polymorphic base at position 240 of the sequence shown in SEQ ID NO. 23, and its base polymorphism is G / C; SNP-11-126349400, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 24, and its base polymorphism is C / T; SNP-11-150316502, which is the polymorphic base at position 238 of the sequence shown in SEQ ID NO. 25, and its base polymorphism is G / A; SNP-11-175529761, which is the polymorphic base at position 255 of the sequence shown in SEQ ID NO. 26, and its base polymorphism is G / C; SNP-12-38432972, which is the polymorphic base at position 227 of the sequence shown in SEQ ID NO. 27, and its base polymorphism is C / G; SNP-12-88588135, which is the polymorphic base at position 253 of the sequence shown in SEQ ID NO. 28, and its base polymorphism is G / A; SNP-13-180557942, which is the polymorphic base at position 282 of the sequence shown in SEQ ID NO. 29, and its base polymorphism is T / C; SNP-14-86914708, which is the polymorphic base at position 224 of the sequence shown in SEQ ID NO. 30, and its base polymorphism is T / C; SNP-14-87149383 is the polymorphic base at position 257 of the sequence shown in SEQ ID NO. 31, and its base polymorphism is G / T.

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