An SNP molecular marker related to wheat pre-harvest sprouting resistance and its application

Abstract

The present invention provides an SNP molecular marker related to wheat pre-harvest sprouting resistance and its application, specifically relating to the technical field of genetic breeding. The SNP locus corresponds to the A / G base mutation at 677353376 bp on chromosome 6B of wheat in the IWGSC RefSeq v1.0 version of the reference genome; a CAPS marker is developed based on the SNP molecular marker, and the CAPS marker is named 6B-4099, and the 6B-4099 has the nucleotide sequence shown in SEQ ID NO.1 or the nucleotide sequence shown in SEQ ID NO.2. The CAPS marker is used to identify wheat varieties resistant to pre-harvest sprouting. The detection method of the present invention is simple and helps to improve the efficiency of molecular breeding for wheat pre-harvest sprouting resistance.

Description

Technical Field

[0001] The present invention belongs to the technical field of wheat genetic breeding, and particularly relates to an SNP molecular marker related to wheat pre-harvest sprouting resistance and its application. Technical Background

[0002] Wheat is prone to the phenomenon of grain sprouting on the ear before harvest, namely pre-harvest sprouting (PHS). PHS not only reduces the wheat yield, but also leads to the deterioration of quality and even the loss of seed value, causing serious economic losses to agricultural production (Xiao et al. 2002). For example, the storage substances (such as proteins, starches, etc.) in the endosperm of wheat affected by PHS will be degraded, so the nutritional quality decreases accordingly, and the seed value is also affected; at the same time, the α-amylase activity in the grains of wheat with PHS increases, resulting in enhanced dough viscosity, poor elasticity of bread and difficult to cut, dark color of steamed buns, enhanced viscosity, poor chewiness of noodles and deteriorated taste (Xiao et al. 2002). It is reported that in the northern part of New South Wales and Queensland in Australia, the PHS disasters caused by summer rainfall affect the quality of 15% of the grains on average every year (Mares 1993). In another important wheat exporting country, Canada, about 8 million hectares of spring wheat, 2 million hectares of durum wheat and 0.4 million hectares of winter wheat are planted, and the quality decline and yield loss caused by PHS occur to a certain extent every year (Clarke et al. 2005).

[0003] Seed dormancy is the main genetic control factor for pre-harvest sprouting resistance. Therefore, breeding wheat varieties with strong seed dormancy and pre-harvest sprouting resistance is the most economical, effective and safe way to cope with pre-harvest sprouting disasters in wheat-growing areas with rainy harvest seasons. Using molecular marker-assisted breeding technology can accelerate the breeding process of new wheat varieties with pre-harvest sprouting resistance. Exploring seed dormancy / pre-harvest sprouting gene loci and developing molecular markers are important prerequisites for carrying out molecular marker-assisted breeding.

[0004] Studies have shown that loci related to wheat seed dormancy / pre-harvest sprouting resistance are distributed across the 21 chromosomes of wheat; some candidate genes have been map-based cloned or homologously cloned, and functional markers have been developed, providing important gene resources and markers for the genetic improvement of wheat pre-harvest sprouting resistance (Anderson et al. 1993; Kato et al. 2001; Osa et al. 2003; Mori et al. 2005; Zhu et al. 2019; Zuo et al. 2019; Zhang et al. 2014, 2017; Yang et al. 2007; Chang et al. 2010; Nakamura et al. 2011; Liu et al. 2013; Torada et al. 2016; Wei et al. 2019). For example, Liu et al. (2008, 2013) detected a major locus QPhs.pseru-3AS on chromosome 3AS, and subsequently mined the candidate gene TaMFT (TaPHS1) through map-based cloning. The positive regulatory effect of TaMFT on PHS resistance was confirmed by means of RNA interference-mediated gene silencing; Zhang et al. (2014, 2017) cloned the homologous gene TaSdr of the rice seed dormancy gene OsSdr4 using comparative genomics methods. TaSdr is located in the second homologous group and is named TaSdr-A1, TaSdr-B1, and TaSdr-D1 respectively; then, CAPS markers (cleaved amplified polymorphism sequence marker, CAPS) Sdr2B and Sdr2A were developed based on the SNPs at -11 upstream of the start codon of TaSdr-B1 and at the 643rd base of TaSdr-A1 respectively, and the variations of these two SNPs were verified to be related to seed dormancy in both linkage populations and natural populations; Bailey et al. (1999) located the gene TaVp-1 encoding the wheat transcription factor VIVIPAROUS-1 on the long arm of the third homologous group by combining comparative genomics and linkage map analysis. In addition, TaPM19-A1 and TaPM19-A1 belonging to the wheat plasma membrane 19 gene family induced by ABA (Barrero et al. 2015), the candidate gene TaMKK3-A encoding mitogen-activated protein kinase 3 (Barrero et al. 2015), and the gene TaQsd1 encoding alanine aminotransferase (Wei et al. 2019) have also been identified to be related to pre-harvest sprouting resistance and seed dormancy. Jing et al. (2021) also confirmed that the Myb10-D transcription factor regulates both wheat grain color and seed dormancy, providing a new idea for the breeding of white-grained wheat varieties resistant to pre-harvest sprouting.

[0005] Although some seed dormancy / PHS resistance genes have been cloned, PHS resistance is a complex quantitative trait controlled by multiple genes, and its genetic regulation mechanism is still unclear at present, which is not conducive to the molecular genetic improvement of wheat pre-harvest sprouting resistance. In addition, the wheat genome is huge and complex, and there are also significant differences in the resistance genetic mechanisms among materials with different genetic backgrounds. Therefore, identifying more PHS resistance gene loci from different germplasm resources and developing molecular markers will help to accelerate the process of molecular design breeding for pre-harvest sprouting resistance. Summary of the Invention

[0006] The object of the present invention is to develop CAPS markers that are easy to detect in large scale batches for a candidate gene 6B-4099 of a new major locus Qphs.ahau-6B controlling PHS resistance, and to be used for molecular marker-assisted breeding of wheat PHS resistance.

[0007] The technical solution is as follows:

[0008] The present invention provides an SNP molecular marker related to wheat pre-harvest sprouting resistance, characterized in that the SNP respectively marks the A / G base mutation at 677353376 bp of chromosome 6B of wheat corresponding to the reference genome IWGSC RefSeq v1.0 version.

[0009] The present invention also provides a CAPS marker developed by using the above SNP molecular marker for identifying wheat varieties resistant to pre-harvest sprouting. The CAPS marker is named 6B-4099, and the nucleotide sequence of 6B-4099 is shown in SEQ ID NO.1 or SEQ ID NO.2.

[0010] The present invention also provides a method for identifying wheat varieties resistant to pre-harvest sprouting by using the above CAPS marker. The specific steps are as follows:

[0011] S1. Design specific primer pairs according to SEQ ID NO.1 and SEQ ID NO.2 of the sequence described in claim 2, and select restriction endonucleases according to the differences between SEQ ID NO.1 and SEQ ID NO.2 of the sequence described in claim 2, so that different cutting results are obtained after SEQ ID NO.1 and SEQ ID NO.2 of the sequence are cut;

[0012] S2. Extract the genomic DNA of the wheat to be detected;

[0013] S3. Perform PCR amplification on the DNA extracted in step S2 with the specific primer pairs designed in step S1 to obtain PCR amplification products;

[0014] S4. Use the restriction endonuclease selected in step S1 to digest the PCR amplification product obtained in step S3, and perform electrophoresis detection on the digested product to determine the wheat variety according to the number of bands.

[0015] Furthermore, the upstream primer in the specific primer designed in step S1 has the nucleotide sequence shown in SEQ ID NO.3, and the downstream primer has the nucleotide sequence shown in SEQ ID NO.4.

[0016] Furthermore, the restriction endonuclease used in step S3 is Hpy188I. If the electrophoresis band of the digested product in step S4 is one, it is a variety susceptible to pre-harvest sprouting; if it is several, it is a variety resistant to pre-harvest sprouting.

[0017] The beneficial effects of the present invention are as follows:

[0018] The present invention develops a CAPS marker for a new major locus Qphs.ahau-6B controlling PHS resistance, and verifies that the CAPS marker 6B-4099 can effectively distinguish wheat varieties with different pre-harvest sprouting resistance levels, providing an important scientific basis for molecular marker-assisted breeding of wheat pre-harvest sprouting resistance. Description of the Drawings

[0019] Figure 1 It is the electrophoresis pattern of using the CAPS marker 6B-4099 to detect wheat varieties Jing 411 (J) and Hongmangchun 21 (H) with different pre-harvest sprouting resistances in the example; among them, J is the band of the J411 type; H is the band of the HMC21 type, and M is the band of the 2K Marker. Detailed Embodiments

[0020] In this example, a new major QTL Qphs.ahau-6B conferring pre-harvest sprouting (PHS) resistance was detected in the Jing 411 / Hongmangchun 21 population using SSR and SLAF markers. The resistant / susceptible DNA pools of this population were scanned with a 660K SNP array, and multiple CAPS markers were developed within the region of the major QTL Qphs.ahau-6B (680-720 Mb), preliminarily determining the authenticity of the major QTL Qphs.ahau-6B. Among them, CAPS marker 6B-4099 was more tightly linked to the major QTL Qphs.ahau-6B. Further, 278 Chinese wheat mini-core germplasm accessions were genotyped with marker 6B-4099, and the Mann-Whitney test (U test) was performed in combination with PHS phenotypes in multiple environments. The results showed that there were significant differences in PHS resistance (P<0.05 or 0.01) between wheat varieties carrying different allelic types of this marker, 6B-4099a (the allele type consistent with Jing 411) and 6B-4099b (the allele type consistent with Hongmangchun 21), verifying that CAPS marker 6B-4099 can effectively distinguish wheat varieties with different PHS resistance levels, providing an important scientific basis for molecular marker-assisted breeding for PHS resistance in wheat. The specific operations are as follows:

[0021] Example

[0022] (I) Determination of germination index (GI) and field sprouting (FS)

[0023] At different times after harvest in 2016-2017 (5 days and 30 days after harvest), 50 seeds with intact embryos were taken from 278 Chinese wheat mini-core germplasm accessions, with 2 replicates. The seeds were evenly placed in a 90-mm-diameter Petri dish with the ventral groove facing down, and 10 ml of sterile water was added. The seeds were cultured for 3 days at 20 °C under a 14-h (day) / 10-h (night) photoperiod. The number of germinated seeds on the first day (n1), the second day (n2), the third day (n3), and the number of ungerminated seeds (n0) after 3 days were recorded. The germination index GI was calculated using the following formula: GI=(3*n1 + 2*n2 + n3) / 3*(n1 + n2 + n3 + n0). The GI of wheat grains 30 days after harvest in 2016 was denoted as 16GI-30d-WHX. The GIs 5 days and 30 days after harvest in 2017 were denoted as 17GI-5d-WHX and 17GI-30d-WHX, respectively.

[0024] FS determination: In 2015 and 2016, there were two natural rainfalls in the field before wheat harvest. 10 spikes were randomly harvested, and then the percentage of germinated grains in all grains was measured, which was recorded as the field sprouting rate (FS). The FS of 278 Chinese wheat mini-core germplasm resources in 2015 and 2016 was measured and recorded as 15FS-WHX and 16FS-WHX.

[0025] (2) Wheat genomic DNA extraction

[0026] The experimental materials for DNA extraction were the parents Jing 411 / Hongmangchun 21 and 278 Chinese wheat mini-core germplasm resources. The specific extraction method was as follows:

[0027] 1. Grind a single wheat grain, take 0.1 g and add it to 0.7 ml of SDS extraction solution (0.1 M Tris-HCl (PH = 8.5), 0.1 M NaCl, 0.05 M EDTA (PH = 8.0), 2% SDS), and lyse at 60 °C for 45 min with multiple oscillations during this period.

[0028] 2. Centrifuge at 12,000 rpm at 4 °C for 10 min.

[0029] 3. Take the supernatant and add an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1), and invert and rotate until the two phases do not quickly separate.

[0030] 4. Centrifuge at 12,000 rpm at 4 °C for 10 min.

[0031] 5. Take the supernatant and add an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1) and invert and rotate several times.

[0032] 6. Centrifuge at 12,000 rpm at 4 °C for 10 min.

[0033] 7. Take the supernatant and add an equal volume of isopropanol, and let it stand in a -20 °C refrigerator for 30 min.

[0034] 8. Centrifuge at 12,000 rpm at 4 °C for 10 min.

[0035] 9. Wash twice with 70% ethanol.

[0036] 10. Centrifuge at 12,000 rpm at 4 °C for 10 min.

[0037] 11. Air-dry the precipitate naturally, store it at 4 °C for later use.

[0038] Obtain the DNA of Jing 411 / Hongmangchun 21 and 278 Chinese wheat mini-core germplasm resources.

[0039] (III) Scanning of Wheat 660K SNP Chip

[0040] Based on the Jing 411 / Hongmangchun 21 recombinant inbred line population, resistant and susceptible PHS mixed pools were constructed, and the wheat 660K chip (Beijing CapitalBio Corporation) was used to scan the resistant / susceptible PHS mixed pools. The resistant and susceptible PHS mixed pools contained 1 resistant PHS parent and 2 extreme resistant PHS mixed pools (5 extreme resistant PHS families were selected and mixed in each resistant PHS bud mixed pool) and 1 susceptible PHS parent and 2 extreme susceptible PHS mixed pools (5 extreme susceptible PHS families were selected and mixed in each susceptible PHS mixed pool), respectively. Excel was used to screen and extract the SNPs that were consistent within the pool and had genotype differences between the pools in the 660K chip. Then, enrichment analysis of the differential SNPs was performed for the major QTL region on chromosome 6B, and the differential SNPs located within the Qphs.ahau-6B target region were selected for the development of CAPS markers.

[0041] (IV) Development of CAPS Marker 6B-4099

[0042] 1. Analysis of Restriction Sites of Differential SNPs

[0043] All SNPs within the target region were analyzed for restriction sites, and it was found that the A / G difference of the SNP AX-108834099 (at 677353376 bp on chromosome 6B) could be distinguished by the restriction endonuclease Hpy188I (New England Biolabs, NEB). The recognition site of Hpy188I is: TCNGA.

[0044] 2. Primer Design

[0045] The flanking sequences of AX-108834099 were BLASTed on the wheat reference genome (IWGSC RefSeq v1.0, website: https: / / urgi.versailles.inra.fr / blast_iwgsc / blast.php) and extended forward and backward to obtain a sequence SEQ ID No.1 or SEQ ID No.2 with a total length of 696 bp.

[0046] Then, using SEQ ID No.1 and SEQ ID No.2 as templates, a pair of specific primers was designed using Primer Premier 5.0 software:

[0047] SEQ ID No.3 6B-4099-F: 5'TGTATCCCAAGGAGCAGT3'

[0048] SEQ ID No.4 6B-4099-R: 5'ATCCCAGGAGTGACGAG 3'

[0049] (5) Amplification, digestion and electrophoresis of the target product

[0050] 1. Preparation of the PCR amplification system: 5 μl of Mix (Vazyme Biotech Co., Ltd), 2.2 μl of sterile water, 0.4 μl of each of the upstream and downstream primers, 100 ng of DNA template (J411 / HMC21 population), and made up to 10 μl with double-distilled water.

[0051] 2. PCR amplification program: Pre-denaturation at 95°C for 5 min; denaturation at 95°C for 15 s; annealing starting from 62°C with a 0.3°C decrease per cycle for 15 s; extension at 72°C for 30 s, 40 cycles; extension at 72°C for 8 min, and incubation at 12°C.

[0052] 3. Digestion system after amplification: 5 μl of PCR product, 1 μl of 10×CutSmart Buffer, 0.5 U of Hpy188I, and made up to 10 μl with double-distilled water.

[0053] 4. Digestion reaction program: 37°C for 8 hours.

[0054] After digestion, 5 μl of the digested product was subjected to typing by 2.5% agarose gel electrophoresis. The band of Jing 411 (susceptible to pre-harvest sprouting) could not be digested, the nucleotide at the corresponding SNP marker was A, and the corresponding genotype was recorded as 6B-4099a, and the nucleotide sequence of the corresponding CAPS marker was SEQ ID NO.1; the band of Hongmangchun 21 (resistant to pre-harvest sprouting) could be cut ( Figure 1 ), the nucleotide at the corresponding SNP marker was G, and the corresponding genotype was recorded as 6B-4099b, and the nucleotide sequence of the corresponding CAPS marker was SEQ ID NO.2.

[0055] Using the above operations, this example also carried out digestion experiments of CAPS markers on Kangxiu 10, Taizhong 23, Jinmai 2148, Dixiuzao, Bima 1, Bima 4, Pingyang 27, Taishan 1, Jinan 2, Youbao, Bainong 3217, Yannong 15, Xinong 6028, Shijiazhuang 407, Jingyang 60, Shite 14, Fuzhuang 30, Shijiazhuang 54, Fengchan 3, Jimai 2, etc. Among them, those that could be cut were Jingyang 60, Shite 14, Fuzhuang 30, Shijiazhuang 54, Fengchan 3 and Jimai 2, and those that could not be cut were Kangxiu 10, Taizhong 23, Jinmai 2148, Dixiuzao, Bima 1, Bima 4, Pingyang 27, Taishan 1, Jinan 2, Youbao, Bainong 3217, Yannong 15, Xinong 6028 and Shijiazhuang 407.

[0056] It shows that according to whether the CAPS marker can be digested by enzymes, it is possible to effectively judge the corresponding wheat pre-harvest sprouting resistance.

[0057] (VI) Verification of CAPS marker 6B-4099 in the natural population

[0058] The results of the Mann-Whitney test (U test) show that (Table 1), in the natural population composed of 278 wheat mini-core germplasms (Table 2), among the wheat varieties carrying two allelic variation types (6B-4099a and 6B-4099b) of the CAPS marker 6B-4099, the pre-harvest sprouting resistance phenotypic values of 15FS-WHX, 16FS-WHX, 16GI-30d-WHX, 17GI-5d-WHX, and 17GI-30d-WHX all reached significant or extremely significant levels, verifying that the CAPS marker 6B-4099 is closely related to pre-harvest sprouting resistance. At the same time, it also proves that this marker can effectively distinguish wheat varieties with different pre-harvest sprouting resistances, providing an important scientific basis for molecular marker-assisted breeding of wheat pre-harvest sprouting resistance.

[0059] Verification of the correlation between CAPS marker 6B-4099 and pre-harvest sprouting resistance using 278 wheat mini-core germplasm materials

[0060]

[0061] Table 1

[0062] Note: ** Indicates a highly significant correlation between the marker and the trait at the 0.01 level; * Indicates a significant correlation between the marker and the trait at the 0.05 level; 6B-4099a is the allelic type consistent with J411, and the nucleotide at the corresponding SNP marker is A; 6B-4099b is the allelic type consistent with HMC21, and the nucleotide at the corresponding SNP marker is G. GI represents the germination index; 15FS-WHX and 16FS-WHX represent the field pre-harvest sprouting rates of the mini-core germplasm materials in 2015 and 2016, respectively; 17GI-5d-WHX represents the GI of the mini-core germplasm harvested in 2017 after 5 days; 16GI-30d-WHX and 17GI-30d-WHX represent the GI of the mini-core germplasm materials harvested in 2016 and 2017 after 5 days, respectively.

[0063] Names of the corresponding varieties of 278 wheat mini-core germplasms

[0064]

[0065]

[0066]

[0067] Table 2

[0068] For a new major effect locus Qphs.ahau-6B that controls pre-harvest sprouting resistance, the present invention developed a CAPS marker 6B-4099 that is closely linked to it. Compared with SSR molecular markers, after electrophoresis detection, the bands of this marker are clear and distinguishable, and the band patterns are significantly different among wheat varieties with different pre-harvest sprouting resistances. The detection method is simple, which helps to improve the targeting and pertinence of molecular marker selection, thereby improving the efficiency of molecular breeding for wheat pre-harvest sprouting resistance.

[0069] As mentioned above, only the preferred specific embodiments of the present invention are described, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, makes equivalent substitutions or changes, and all should be covered within the protection scope of the present invention. Sequence Listing <110> Anhui Agricultural University <120> An SNP Molecular Marker Related to Wheat Pre-Harvest Sprouting Resistance and Its Application <160> 4 <170> SIPOSequenceListing 1.0 <210> 1 <211> 696 <212> DNA <213> Triticum aestivum L <400> 1 gacatctgca aaaccgatga tagatagttt cctcatgccc gcaagccaca caaaacacgc 60 caggttttat ccttcgccta tgtagctctg atcctactac gagtccaatc aaaatgagcc 120 tccacaaatg tatctttgca tttcctggta attttgtatc ccaaggagca gtgtatcctt 180 tatgcttact caccgaggtc gaagactccg gctgtccgga tcttgcttgt ttcatcccca 240 ttttgagatg ataggctgac ctcaccgtga agatgccatt ctttgtaaag ttccaggctt 300 ggtagtcctc cacccccacc ctgccaacaa caatctgcaa gatatctaaa gcattatctg 360 gtgaaaacat ctccataact ctctgaaggt tccatgtgct gccatccgga gttagaaggt 420 ccgctaccct cgtcactcct gggatgaaaa tttgccccaa gggcttcatt gatccacttc 480 ttagaatcca atttgcatgg tggatgttca ctcttgaccc gtctccaatt ctccacacca 540 gcccctccct caagaggtct cggccatgca gaatactttt gaaagtgtat gaagctgtag 600 atgggcacgt agcattcata atagaatctt gtttgaaata gtgttccctg aggactctcg 660 cacacaaata ctctggacac tgcaagatct gccaag 696 <210> 2 <211> 696 <212> DNA <213> Triticum aestivum L <400> 2 gacatctgca aaaccgatga tagatagttt cctcatgccc gcaagccaca caaaacacgc 60 caggttttat ccttcgccta tgtagctctg atcctactac gagtccaatc aaaatgagcc 120 tccacaaatg tatctttgca tttcctggta attttgtatc ccaaggagca gtgtatcctt 180 tatgcttact caccgaggtc gaagactccg gctgtccgga tcttgcttgt ttcatcccca 240 ttttgagatg ataggctgac ctcaccgtga agatgccatt ctttgtaaag ttccaggctt 300 ggtagtcctc cacccccacc ctgccaacaa caatctgcaa gatatctgaa gcattatctg 360 gtgaaaacat ctccataact ctctgaaggt tccatgtgct gccatccgga gttagaaggt 420 ccgctaccct cgtcactcct gggatgaaaa tttgccccaa gggcttcatt gatccacttc 480 ttagaatcca atttgcatgg tggatgttca ctcttgaccc gtctccaatt ctccacacca 540 gcccctccct caagaggtct cggccatgca gaatactttt gaaagtgtat gaagctgtag 600 atgggcacgt agcattcata atagaatctt gtttgaaata gtgttccctg aggactctcg 660 cacacaaata ctctggacac tgcaagatct gccaag 696 <210> 3 <211> 18 <212> DNA <213> Synthetic (Synotype) <400> 3 tgtatcccaa ggagcagt 18 <210> 4 <211> 17 <212> DNA <213> Synthetic (Synotype) <400> 4 atcccaggag tgacgag 17

Claims

1. A CAPS marker developed using SNP molecular markers for identifying pre-harvest sprouting resistance in wheat, characterized in that, The SNP molecular marker corresponds to an A / G base mutation at 677353376 bp on chromosome 6B of wheat in the IWGSC RefSeq v1.0 version of the reference genome. The CAPS marker is named 6B-4099, and the nucleotide sequence of 6B-4099 is shown in SEQ ID NO.1 or SEQ ID NO.

2.

2. A method for identifying wheat varieties resistant to pre-harvest sprouting using the CAPS markers described in claim 1, characterized in that, The specific steps are as follows: S1. Design a pair of specific primers according to the sequence SEQ ID NO.1 and the sequence SEQ ID NO.2 in claim 1. Select a restriction endonuclease based on the differences between SEQ ID NO.1 and SEQ ID NO.2 according to claim 1, such that different cleavage results are obtained after SEQ ID NO.1 and SEQ ID NO.2 are cleaved; wherein, the selected restriction endonuclease is Hpy188I ; S2. Extract the genomic DNA of the wheat to be detected. S3. Perform PCR amplification on the DNA extracted in step S2 using the pair of specific primers designed in step S1 to obtain a PCR amplification product. S4. Use the restriction endonuclease selected in step S1 to digest the PCR amplification product obtained in step S3, and perform electrophoresis detection on the digestion product. If there is one electrophoresis band, it is a variety susceptible to pre-harvest sprouting; if there are several electrophoresis bands, it is a variety resistant to pre-harvest sprouting.

3. The method for identifying wheat varieties resistant to pre-harvest sprouting by CAPS markers according to claim 2, characterized in that, The nucleotide sequence of the upstream primer in the pair of specific primers designed in step S1 is shown in SEQ ID NO.3, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.4.

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