Wheat scab-resistant gene KASP marker and application thereof

Abstract

The invention relates to the technical field of wheat breeding, in particular to a wheat gibberellic disease resistance gene KASP marker and application thereof, the wheat gibberellic disease resistance gene KASP marker mainly comprises DQK009, DQK011, DQK004, DQK008, DQK005, DQK007, DQK001 and DQK002, and the technical problem that at present, high-throughput screening is difficult to conduct on other uncloned genes of wheat is mainly solved. The invention further provides application of the KASP marker in two aspects of wheat genotype detection and rapid and efficient breeding of the gibberellic disease resistant wheat, and the technical problem that the breeding time of the gibberellic disease resistant wheat is long at present is mainly solved.

Description

Technical Field

[0001] This application relates to the field of wheat breeding technology, specifically to a wheat scab resistance gene KASP marker and its application. Background Technology

[0002] Wheat (Triticum asetivum L.) is the most widely distributed, largest-planted, and highest-yielding grain crop in the world, and also one of the most important grain crops in my country. In my country, Fusarium head blight has become the primary disease restricting high and stable wheat yields. Especially affected by global warming and changes in farming systems in recent years, wheat Fusarium head blight has gradually spread northward and westward.

[0003] Fusarium head blight (FHB) is a fungal disease that seriously affects wheat yield and quality, primarily caused by various Fusarium fungi belonging to the genus *Fusarium*. In addition, infected grains contain multiple toxins secreted by *Fusarium*, such as deoxynivalenol (DON) and zearalenone (ZEN). Excessive consumption of contaminated wheat products can lead to poisoning symptoms such as vomiting, nausea, dizziness, and diarrhea, seriously endangering the health of humans and livestock. *Fusarium* primarily infects the wheat ear during the flowering stage, resulting in a very narrow window for chemical control and making pesticide application difficult. Therefore, improving the resistance of wheat varieties to Fusarium head blight through genetic modification is the most economical and effective measure to control the occurrence and spread of this disease.

[0004] Wheat scab resistance is primarily a quantitative trait controlled by multiple genes and is highly susceptible to environmental influences. To date, nine major genes have been formally named, but only the scab resistance genes Fhb1 and Fhb7 have been cloned and can be accurately detected using diagnostic markers. However, the remaining uncloned genes can only be identified using gene-linked SSR markers, making high-throughput gene screening difficult. Furthermore, in variety breeding, traditional methods for developing disease-resistant varieties are time-consuming, and the efficiency and throughput of SSR markers used in marker-assisted selection of scab resistance genes are relatively low. Summary of the Invention

[0005] The problem this application aims to solve is to provide a KASP marker for wheat resistance to Fusarium head blight and its application, in order to address the current technical issues of difficulty in high-throughput screening of other uncloned wheat genes and the long breeding time required for Fusarium head blight-resistant wheat.

[0006] To solve the above-mentioned technical problems, this application adopts the following technical solution:

[0007] On one hand, this application provides a KASP marker for wheat resistance to Fusarium head blight, including a KASP marker DQK009 for SNP1, the sequences of the competitive primers for DQK009 are shown in SEQ ID NO.1 and SEQ ID NO.2, and the nucleotide sequence of the common primer for DQK009 is shown in SEQ ID NO.3; a KASP marker DQK011 for SNP2, the sequences of the competitive primers for DQK008 are shown in SEQ ID NO.4 and SEQ ID NO.5, and the nucleotide sequence of the common primer for DQK011 is shown in SEQ ID NO.6; a KASP marker DQK004 for SNP3, the sequences of the competitive primers for DQK004 are shown in SEQ ID NO.7 and SEQ ID NO.8, and the nucleotide sequence of the common primer for DQK004 is shown in SEQ ID NO.9; and a KASP marker DQK008 for SNP4, the sequences of the competitive primers for DQK008 are shown in SEQ ID NO.10 and SEQ ID NO.2. As shown in NO.11, the nucleotide sequence of the common primer for DQK008 is shown in SEQ ID NO.12; for the KASP marker DQK005 targeting SNP5, the sequences of the competing primers for DQK005 are shown in SEQ ID NO.13 and SEQ ID NO.14, and the nucleotide sequence of the common primer for DQK005 is shown in SEQ ID NO.15; for the KASP marker DQK007 targeting SNP6, the sequences of the competing primers for DQK007 are shown in SEQ ID NO.16 and SEQ ID NO.17, and the nucleotide sequence of the common primer for DQK007 is shown in SEQ ID NO.18; for the KASP marker DQK001 targeting SNP7, the sequences of the competing primers for DQK009 are shown in SEQ ID NO.19 and SEQ ID NO.20, and the nucleotide sequence of the common primer for DQK001 is shown in SEQ ID NO. As shown in NO.21; for the KASP marker DQK002 for SNP8, the sequences of the competitive primers for DQK002 are shown in SEQ ID NO.22 and SEQ ID NO.23, and the nucleotide sequence of the common primer for DQK002 is shown in SEQ ID NO.24.

[0008] In some embodiments, the SNP sites for the development of the wheat Fusarium head blight resistance gene KASP marker include: SNP1, located at chromosome position chr6B:118879886, the nucleotide sequence of which is shown in SEQ ID NO.25; SNP2, located at chromosome position chr6B:152096128, the nucleotide sequence of which is shown in SEQ ID NO.26; SNP3, located at chromosome position chr6B:165725747, the nucleotide sequence of which is shown in SEQ ID NO.27; SNP4, located at chromosome position chr6B:169959743, the sequence of which is shown in SEQ ID NO.28; SNP5, located at chromosome position chr4B:463122524, the nucleotide sequence of which is shown in SEQ ID NO.29; and SNP6, located at chromosome position chr4B:545511172, the nucleotide sequence of which is shown in SEQ ID NO.25. As shown in NO.30; SNP7, located at chromosome position chr5A:191466469, has a nucleotide sequence as shown in SEQ ID NO.31; SNP8, located at chromosome position chr5A:209001461, has a nucleotide sequence as shown in SEQ ID NO.32.

[0009] In some embodiments, wheat with the GG genotype of DQK009 carries the disease resistance gene Fhb2; wheat with the CC genotype of DQK009 does not carry the disease resistance gene Fhb2; wheat with the GG genotype of DQK011 carries the disease resistance gene Fhb2; wheat with the AA genotype of DQK011 does not carry the disease resistance gene Fhb2; wheat with the TT genotype of DQK004 carries the disease resistance gene Fhb2; wheat with the CC genotype of DQK004 does not carry the disease resistance gene Fhb2; wheat with the TT genotype of DQK008 carries the disease resistance gene Fhb2; and wheat with the GG genotype of DQK008 does not carry the disease resistance gene Fhb2. Wheat with genotype AA (Fhb2); wheat with genotype GG (DQK005) does not carry the disease resistance gene Fhb4; wheat with genotype TT (DQK007) carries the disease resistance gene Fhb4; wheat with genotype CC (DQK007) does not carry the disease resistance gene Fhb4; wheat with genotype TT (DQK001) carries the disease resistance gene Fhb5; wheat with genotype CC (DQK001) does not carry the disease resistance gene Fhb5; wheat with genotype TT (DQK002) carries the disease resistance gene Fhb5; wheat with genotype CC (DQK002) does not carry the disease resistance gene Fhb5.

[0010] On the other hand, the present invention provides an application of the wheat scab resistance gene KASP marker in detecting wheat genotypes.

[0011] In some embodiments, the method for detecting wheat genotype includes the following steps: S1, using the genomic DNA of the wheat to be tested as a template, performing PCR amplification using competitive primers DQK011, DQK004, DQK008, DQK005, DQK007, DQK001 or DQK002; S2, performing fluorescence detection on the PCR product obtained in S1, and determining the genotype of the wheat to be tested based on the obtained fluorescence signal.

[0012] On the other hand, this invention provides an application of the wheat scab resistance gene KASP marker in rapid and efficient breeding of scab-resistant wheat.

[0013] In some embodiments, the above-mentioned rapid and efficient breeding method for wheat resistant to Fusarium head blight includes the following steps: S1, selecting resistant materials containing 3-4 of the Fusarium head blight resistance genes Fhb1, Fhb2, Fhb4, and Fhb5 as the male parent, and selecting materials with excellent agronomic traits from local varieties as the female parent, and performing hybridization; using the F1 hybrid plants as the female parent and materials with excellent agronomic traits as the recurrent male parent, performing backcrossing 3 times to obtain the BC3F1 generation; S2, the BC3F1 generation continues to propagate rapidly to obtain the BC3F2 generation, and planting BC3F2 in the field during the normal season, sampling individual plants with tags to extract genomic DNA, using DQK011, DQK004, DQK008, DQK005, DQK007, DQK001, and DQK002 for genotyping, identifying the disease resistance genes carried by individual plants, obtaining individual plants carrying multiple Fusarium head blight resistance genes, and harvesting and threshing them to obtain BC3F2. F3 generation; BC3F3 generation plants were harvested and planted individually in rows during the peak season. Genomic DNA was extracted from each plant and tagged. Genotyping was performed using DQK011, DQK004, DQK008, DQK005, DQK007, DQK001, and DQK002 to identify disease resistance genes carried by individual plants. Homozygous plants carrying multiple Fusarium head blight resistance genes were obtained. Fusarium head blight resistance was then assessed by drip inoculation of individual flowers during the flowering period. Disease resistance: Harvest single plants carrying multiple homozygous resistance genes to Fusarium head blight and threshing to obtain BC3F4 generation; S3: Continue field propagation using BC3F4 generation, harvest by line, and successively enter the identification nursery and variety comparison nursery to select stable lines with good resistance to Fusarium head blight and excellent agronomic traits; The obtained stable lines with good resistance to Fusarium head blight and excellent agronomic traits are put into multi-point trials, and lines with excellent resistance to Fusarium head blight and excellent agronomic traits are selected to enter regional trials, thus obtaining Fusarium head blight resistant wheat.

[0014] In some embodiments, in S1, the light and temperature for hybridization and backcrossing are controlled at 17-19°C with 3.5-4.5 hours of darkness and 21-23°C with 19.5-20.5 hours of light.

[0015] In some embodiments, the native strain includes the Sichuan strain. Wheat from other regions may also be selected depending on the specific region.

[0016] In some embodiments, excellent agronomic traits include plant height of 80-90cm, compact plant type, 3-5 effective ears per plant, early maturity, and high yield.

[0017] This application has the following beneficial effects:

[0018] (1) This invention obtains genotyping data of native Sichuan wheat lines and Fusarium head blight-resistant parents through sequencing. Based on the genetic mapping regions and physical locations of the Fusarium head blight resistance genes Fhb2, Fhb4, and Fhb5, core SNPs showing differences between parents were identified. KASP markers for Fusarium head blight resistance genes were designed, and their specificity was verified. The KASP markers of this invention enable rapid, efficient, and high-throughput identification of Fusarium head blight resistance gene types. This allows for efficient detection and tracking of Fusarium head blight resistance genes in wheat varieties / lines and breeding populations, facilitating the efficient application of Fusarium head blight resistance genes in wheat breeding.

[0019] (2) This invention utilizes KASP marker identification technology combined with plant growth chamber rapid propagation and limited backcrossing breeding methods to purposefully improve the resistance of native parents to Fusarium head blight, which greatly shortens the traditional breeding cycle, reduces breeding costs, and accelerates the breeding application of Fusarium head blight resistance genes. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in this disclosure, the accompanying drawings used in some embodiments of this disclosure will be briefly described below. Obviously, the drawings described below are merely drawings of some embodiments of this disclosure, and those skilled in the art can obtain other drawings based on these drawings. Furthermore, the drawings described below can be regarded as schematic diagrams and are not intended to limit the actual process of the methods involved in the embodiments of this disclosure.

[0021] Figure 1 This is a KASP marker typing diagram for the Fusarium head blight resistance gene in this invention, where DQK009, DQK011, DQK004, and DQK008 are detection markers for the Fhb2 gene, DQK005 and DQK007 are detection markers for the Fhb4 gene, and DQK001 and DQK002 are detection markers for the Fhb5 gene.

[0022] Figure 2This is a flowchart of the rapid breeding technology for Fusarium head blight resistance genes using KASP marker-assisted selection in this invention;

[0023] Figure 3 This is a diagram showing the growth of wheat in a plant growth chamber. Detailed Implementation

[0024] The technical solutions in some embodiments of this disclosure will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. Based on the embodiments provided in this disclosure, all other embodiments obtained by those skilled in the art are within the scope of protection of this disclosure.

[0025] 1. Plant materials

[0026] The 17 materials used for 100K SNP microarray sequencing included 14 main Sichuan varieties and reserve lines: 21Pin3-9, 21Pin4-3, 21Pin6-5, Chuanmai 614, Chuanmai 618, Chuanmai 620, Chuanmai 623, Chuanmai 625, Chuanmai 703, Chuanmai 704, Chuanmai 914, Mianmai 60, Neimai 175, and Shumai 1925. Also included were 3 Fusarium head blight resistant germplasms from Nanjing Agricultural University: NMAS069, NMAS070, and NMAS071, with genotypes of Zhoumai 27 (Fhb1, Fhb2, Fhb4), Bainong 4199 (Fhb1, Fhb4, Fhb5), and Chuanmai 64 (Fhb1, Fhb2, Fhb4, Fhb5), respectively.

[0027] 2. SNP screening between native wheat varieties and Fusarium head blight resistant parents

[0028] To obtain the Special Numerous NPs (SNPs) between native Sichuan wheat lines and Fusarium head blight-resistant parents, genotyping data of these two genes were obtained using 100K SNP microarray sequencing. All SNP data underwent quality control and filtering, and gene mapping interval SNP analysis was performed between 14 Sichuan wheat varieties (lines) and 3 Fusarium head blight-resistant parents. Based on the genetic linkage maps of the Fhb2, Fhb4, and Fhb5 genes, the physical locations of the linkage and co-segregation markers of the Fusarium head blight resistance genes Fhb2, Fhb4, and Fhb5 on the chromosomes were determined. Based on the SNP information and the physical locations of the markers on the chromosomes, SNPs between Sichuan wheat lines and Fusarium head blight-resistant parents within the co-segregation intervals with the resistance genes were obtained. The co-segregating mapping region of the disease resistance gene Fhb5 contains many core SNPs between the disease resistance parent and Sichuan varieties (lines), while the co-segregating mapping regions of the disease resistance genes Fhb2 and Fhb4 do not contain any core SNPs between the disease resistance parent and Sichuan varieties (lines). Therefore, further analysis was conducted within the linkage mapping regions of the disease resistance genes Fhb2 and Fhb4 to obtain core SNPs between the disease resistance parent and Sichuan varieties (lines) for the design of KASP markers.

[0029] 3. Development and identification of KASP markers for Fusarium head blight resistance genes Fhb2, Fhb4, and Fhb5

[0030] Eight key SNP loci (see Table 1) were obtained through SNP screening between Sichuan wheat varieties (lines) and resistant parents within the linkage or co-segregation intervals of Fusarium head blight resistance genes. KASP markers specific to the resistance genes were designed using the upstream and downstream sequences of these key SNP loci. KASP marker primers were synthesized by Sangon Biotech Co., Ltd. Four pairs of KASP primers were designed based on the key SNP information to identify the resistance gene Fhb2, located flanking the Fhb2 interval. DQK004 and DQK008 were used to detect the Fhb2 gene carried in the resistant parents NMAS071 and NMAS069, respectively. Simultaneously, two pairs of KASP primers were designed for the resistance genes Fhb4 and Fhb5, respectively. DQK005 and DQK007 were located flanking the Fhb4 interval. DQK005 detected the Fhb4 gene carried in NMAS069, while DQK007 could simultaneously detect the Fhb4 gene carried in all three resistant parents. The markers DQK001 and DQK002 are located on either side of the Fhb5 region of the disease resistance gene, and can simultaneously detect the Fhb5 gene carried in three disease-resistant parents.

[0031] Table 1. Information on some core SNPs within the linkage regions of the disease resistance genes Fhb2, Fhb4, and Fhb5.

[0032]

[0033]

[0034]

[0035] Detailed information on the KASP marker primers is shown in Table 2. To verify the specificity of the eight KASP primer pairs, primer amplification was performed using 17 wheat varieties (lines) with known genotypes. Three Fusarium head blight-resistant parents were used as positive controls, 14 Sichuan wheat varieties (lines) as negative controls, and ddH2O as a blank control. The KASP reaction system consisted of 10 μL of 10 μL of reaction mixture, including 2 μL of DNA dilution buffer (20-100 ng / μL), 1.4 μL of mixed primers, 1.6 μL of double-distilled water, and 5 μL of 2×KASP reaction mixture (Beijing Jiacheng Biotechnology Co., Ltd.). ABI Veriti was used. TM A 96-gene amplification instrument was used, employing a touch-down PCR amplification program. The specific steps were as follows: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 20 s; annealing / extension at 61℃-55℃ for 40 s, 10 cycles, with a temperature decrease of 0.6℃ per cycle; 95℃ denaturation for 20 s; 55℃ annealing / extension for 40 s, 33 cycles. PCR amplification products were detected for fluorescence at 35℃ for 30 s using an ABI QuantStudio™ 6 Flex real-time fluorescence instrument, and genotyping was determined based on the fluorescence signal. The genotyping results of the 8 KASP markers were consistent with the SNP results, confirming the feasibility of using the 8 KASP primers as molecular marker-assisted detection.

[0036] Table 2. Primer sequence information for KASP markers of disease resistance genes.

[0037]

[0038]

[0039] 4. Rapid and efficient breeding applications of Fusarium head blight resistance genes Fhb2, Fhb4, and Fhb5

[0040] In the plant growth chamber, light and temperature were controlled at 18℃ for 4 hours in darkness and 22℃ for 20 hours in light. Sexual hybridization was performed using the Sichuan native strain Mianmai 60 as the female parent and the introduced Fusarium head blight resistant material NMAS071 as the male parent to obtain hybrid seeds. Using the F1 generation hybrids as the female parent and Mianmai 60 as the recurrent male parent, three backcrosses were conducted to obtain BC3F1 generation seeds. These seeds were then rapidly propagated in the plant growth chamber to obtain BC3F2 generation seeds.

[0041] BC3F2 was planted in the field during the normal season and planted individually, numbered 1-90. Genomic DNA was extracted from each plant. The 90 BC3F2 plants were genotyped using the known KASP marker TaHRC-Kasp for the Fhb1 gene, the KASP markers DQK004 and DQK009 for the Fhb2 gene (resistance to Fusarium head blight), the KASP markers DQK005 and DQK007 for the Fhb4 gene, and the KASP markers DQK001 and DQK002 for the Fhb5 gene. NMAS071 was used as a positive control, Mianmai 60 as a negative control, and ddH2O as a blank control. A 10 μL KASP reaction system was used to screen and identify the resistance genes Fhb1, Fhb2, Fhb4, and Fhb5 in the 90 BC3F2 plants. Three plants carrying resistance genes Fhb1 and Fhb2, one plant carrying resistance genes Fhb1, Fhb2, and Fhb5, and one plant carrying resistance genes Fhb2, Fhb4, and Fhb5 were obtained. Individual plants carrying multiple Fusarium head blight resistance genes were harvested and threshed to obtain the BC3F3 generation. The harvested BC3F3 generation plants were planted individually in rows during the regular season. From each row, 50 plants were sampled and tagged for genomic DNA extraction. KASP markers were used to identify plants carrying homozygous resistance genes. Plants with homozygous resistance genes were inoculated with Fusarium head blight at flowering time to assess Fusarium head blight resistance. At least two plants with homozygous resistance genes and exhibiting Fusarium head blight resistance were obtained from each line.

[0042] The homozygous disease-resistant BC3F4 generation obtained above was propagated by generation in the summer of the same year, and harvested by line. The BC3F5 generation was successively sent to the identification nursery and the comparison nursery. During the seedling stage, the emergence, tillering ability, cold and drought resistance of each line were observed. During the flowering stage, Fusarium head blight was inoculated to identify Fusarium head blight resistance. During the grain-filling stage, disease resistance, lodging resistance and other traits were investigated. At maturity, the three yield factors, namely the number of ears per mu, the number of grains per ear, and the thousand-grain weight, were measured, and the yield was determined after harvest. Superior lines with compact plant type, good disease resistance and high yield were selected to enter the multi-location trial the following year. Superior lines with strong adaptability, good disease resistance and high yield were continued to be submitted to the regional trial the following year.

[0043] The above description is merely a preferred embodiment of this disclosure. It should be understood that this disclosure is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or techniques or knowledge in related fields. Any modifications and variations made by those skilled in the art that do not depart from the spirit and scope of this disclosure should be within the protection scope of the appended claims.

Claims

1. A KASP marker for wheat resistance to Fusarium head blight, characterized in that, include: For the KASP marker DQK009 for SNP1, the sequences of the competitive primers for DQK009 are shown in SEQ ID NO.1 and SEQ ID NO.2, and the nucleotide sequence of the common primer for DQK009 is shown in SEQ ID NO.3; For the KASP marker DQK011 of SNP2, the sequences of the competing primers of DQK008 are shown in SEQ ID NO.4 and SEQ ID NO.5, and the nucleotide sequence of the common primer of DQK011 is shown in SEQ ID NO.6; For the KASP marker DQK004 targeting SNP3, the sequences of the competitive primers for DQK004 are shown in SEQ ID NO.7 and SEQ ID NO.8, and the nucleotide sequence of the common primer for DQK004 is shown in SEQ ID NO.9; For the KASP marker DQK008 for SNP4, the sequences of the competitive primers for DQK008 are shown in SEQ ID NO.10 and SEQ ID NO.11, and the nucleotide sequence of the common primer for DQK008 is shown in SEQ ID NO.12; For the KASP marker DQK005 for SNP5, the sequences of the competitive primers for DQK005 are shown in SEQ ID NO.13 and SEQ ID NO.14, and the nucleotide sequence of the common primer for DQK005 is shown in SEQ ID NO.15; For the KASP marker DQK007 for SNP6, the sequences of the competitive primers for DQK007 are shown in SEQ ID NO.16 and SEQ ID NO.17, and the nucleotide sequence of the common primer for DQK007 is shown in SEQ ID NO.18; For the KASP marker DQK001 for SNP7, the sequences of the competing primers for DQK009 are shown in SEQ ID NO.19 and SEQ ID NO.20, and the nucleotide sequence of the common primer for DQK001 is shown in SEQ ID NO.21; For the KASP marker DQK002 for SNP8, the sequences of the competitive primers for DQK002 are shown in SEQ ID NO.22 and SEQ ID NO.23, and the nucleotide sequence of the common primer for DQK002 is shown in SEQ ID NO.

24.

2. The wheat Fusarium head blight resistance gene KASP marker according to claim 1, characterized in that, The SNP sites used for the development of KASP markers for wheat resistance to Fusarium head blight include: The SNP1 is located at chromosome position chr6B:118879886, and the nucleotide sequence at this position is shown in SEQ ID NO.

25. The SNP2 is located at chromosome position chr6B:152096128, and the nucleotide sequence at this position is shown in SEQ ID NO.26; The SNP3 is located at chromosome position chr6B:165725747, and the nucleotide sequence at this position is shown in SEQ ID NO.

27. The SNP4 is located at chromosome position chr6B:169959743, and the sequence at this position is shown in SEQ ID NO.28; The SNP5 is located at chromosome position chr4B:463122524, and the nucleotide sequence at this position is shown in SEQ ID NO.29; The SNP6 is located at chromosome position chr4B:545511172, and the nucleotide sequence at this position is shown in SEQ ID NO.30; The SNP7 is located at chromosome position chr5A:191466469, and the nucleotide sequence at this position is shown in SEQ ID NO.31; The SNP8 is located at chromosome position chr5A:209001461, and the nucleotide sequence at this position is shown in SEQ ID NO.

32.

3. The wheat Fusarium head blight resistance gene KASP marker according to claim 1, characterized in that, The wheat with the GG genotype of DQK009 carries the disease resistance gene Fhb2; the wheat with the CC genotype of DQK009 does not carry the disease resistance gene Fhb2. The wheat with the GG genotype of DQK011 carries the disease resistance gene Fhb2; the wheat with the AA genotype of DQK011 does not carry the disease resistance gene Fhb2. The wheat with the TT genotype of DQK004 carries the disease resistance gene Fhb2; the wheat with the CC genotype of DQK004 does not carry the disease resistance gene Fhb2. The wheat with the TT genotype of DQK008 carries the disease resistance gene Fhb2; the wheat with the GG genotype of DQK008 does not carry the disease resistance gene Fhb2. The wheat with the AA genotype of DQK005 carries the disease resistance gene Fhb4; the wheat with the GG genotype of DQK005 does not carry the disease resistance gene Fhb4. The wheat with the TT genotype of DQK007 carries the disease resistance gene Fhb4; the wheat with the CC genotype of DQK007 does not carry the disease resistance gene Fhb4. The wheat with the TT genotype of DQK001 carries the disease resistance gene Fhb5; the wheat with the CC genotype of DQK001 does not carry the disease resistance gene Fhb5. The wheat with the TT genotype of DQK002 carries the disease resistance gene Fhb5; the wheat with the CC genotype of DQK002 does not carry the disease resistance gene Fhb5.

4. The application of the wheat scab resistance gene KASP marker as described in any one of claims 1-3 in the detection of wheat genotypes.

5. The application according to claim 4, characterized in that, The method for detecting wheat genotypes includes the following steps: S1. Using the genomic DNA of the wheat to be tested as a template, PCR amplification is performed using the competitive primers DQK011, DQK004, DQK008, DQK005, DQK007, DQK001 or DQK002. S2. Perform fluorescence detection on the PCR products obtained in S1, and determine the genotype of the wheat to be tested based on the obtained fluorescence signal.

6. The application of the wheat Fusarium head blight resistance gene KASP marker as described in any one of claims 1-3 in rapid and efficient breeding of Fusarium head blight resistant wheat.

7. The application according to claim 6, characterized in that, Rapid and efficient breeding methods for wheat resistant to Fusarium head blight include the following steps: S1. Select disease-resistant materials that aggregate 3-4 genes from the Fusarium head blight resistance genes Fhb1, Fhb2, Fhb4, and Fhb5 as the male parent, and select materials with excellent agronomic traits from local strains as the female parent, and perform hybridization; use the F1 hybrid plants as the female parent and the materials with excellent agronomic traits as the recurrent male parent, and perform backcrossing 3 times to obtain the BC3F1 generation; S2 and BC3F1 generations continued to propagate rapidly, obtaining BC3F2 generation. BC3F2 plants were sown in the field during the normal season. Genomic DNA was extracted from individual plants by labeling them. Genotyping was performed using DQK011, DQK004, DQK008, DQK005, DQK007, DQK001, and DQK002 to identify disease resistance genes carried by individual plants. Individual plants carrying multiple Fusarium head blight resistance genes were obtained, and these were harvested and threshed to obtain BC3F3 generation. The harvested BC3F3 generation plants were then planted in the field during the normal season. Individual plants were planted in rows, and each plant was tagged and sampled for genomic DNA extraction. Genotyping was performed using the DQK011, DQK004, DQK008, DQK005, DQK007, DQK001, and DQK002 genes to identify the disease resistance genes carried by the individual plants. Homozygous plants carrying multiple Fusarium head blight resistance genes were obtained. Fusarium head blight resistance was assessed by drip inoculation of individual flowers during the flowering period. The individual plants carrying multiple homozygous Fusarium head blight resistance genes and resistant to Fusarium head blight were harvested and threshed to obtain the BC3F4 generation. S3. Continue field propagation using the BC3F4 generation, harvest by line, and sequentially enter the identification nursery and variety comparison nursery to select stable lines with good resistance to Fusarium head blight and excellent agronomic traits; enter the obtained stable lines with good resistance to Fusarium head blight and excellent agronomic traits into multi-point trials, and select lines with excellent agronomic traits to enter regional trials, thus obtaining the Fusarium head blight resistant wheat.

8. The application according to claim 7, characterized in that, In step S1, the light and temperature for hybridization and backcrossing are controlled at 17-19℃ with 3.5-4.5 hours of darkness and 21-23℃ with 19.5-20.5 hours of light.

9. The application according to claim 7, characterized in that, The local strains include the Sichuan strains.

10. The application according to claim 7, characterized in that, The excellent agronomic traits include a plant height of 80-90cm, compact plant type, 3-5 effective ears per plant, early maturity, and high yield.

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