Molecular Markers for Wheat Stripe Rust Resistance Loci and Their Applications

Abstract

The present application discloses molecular markers for wheat stripe rust resistance loci and their applications, belonging to the technical fields of molecular biology and crop breeding. The technical problem to be solved by this application is: how to screen wheat with stripe rust resistance. For this purpose, the present application provides a method for identifying or assisting in the identification of wheat stripe rust resistance, and the method includes detecting the genotype of the AX-109906455 locus in the wheat genome; the AX-109906455 locus is a SNP locus in the wheat genome, such as the 37th nucleotide of SEQ ID No. 4, and the nucleotide type thereof is G or C. The test wheat with the CC genotype at the AX-109906455 locus has wheat stripe rust resistance. The method provided by the present application can be used for molecular marker-assisted selection breeding, providing material reserves and technical support for the genetic improvement of wheat stripe rust resistance traits.

Description

Technical Field

[0001] This application belongs to the technical fields of molecular biology and crop breeding, and particularly relates to molecular markers for wheat stripe rust resistance loci and their applications. Background Art

[0002] Wheat stripe rust is an air-borne fungal disease caused by Puccinia striiformis West. f. sp. tritici Eriks., which has a fast spreading speed and can infect the leaves and above-ground vegetative organs of wheat, such as leaf sheaths, spikes, culms, glumes, and awns, etc. In China, except for the spring wheat region in the Northeast, other wheat planting areas may be affected by stripe rust, especially in key planting areas such as the Northwest, Southwest, and Huang-Huai-Hai regions, where the disease occurs frequently and has a serious impact. Therefore, preventing and controlling wheat stripe rust is an important task. Although field management measures such as spraying fungicides, adjusting planting density, optimizing population structure, and fertilization can partially alleviate the disease, chemical control methods are costly and pollute the environment. Cultivating stripe rust resistant varieties is an economical, safe, and effective solution.

[0003] Currently, scientists have identified and named approximately 90 stripe rust resistant genes, which are distributed on 20 out of 21 chromosomes. However, due to the relatively fast variation of stripe rust races, some resistance genes are prone to loss, and some cloned resistance genes are linked to undesirable traits, resulting in a limited number of effective genes available for resistance improvement. Therefore, exploring new disease resistance loci (QTL) and developing available molecular markers have become the focus of disease resistance breeding work. Lantian 25 selected by the Wheat Research Institute of Lanzhou University of Commerce has a relatively good level of stripe rust resistance. Summary of the Invention

[0004] The technical problem to be solved by this application is: how to screen wheat with stripe rust resistance.

[0005] To solve the above technical problem, this application provides a method for identifying or assisting in the identification of wheat stripe rust resistance, the method comprising using a substance for detecting the genotype of locus AX-109906455 to detect the genotype of the wheat to be tested, and identifying or assisting in the identification of wheat stripe rust resistance according to the genotype of locus AX-109906455 of the wheat to be tested;

[0006] The locus AX-109906455 is a SNP locus in the wheat genome, which is the 37th nucleotide of SEQ ID No. 4, and its nucleotide type is G or C.

[0007] The genotype of the AX-109906455 locus can be CC, GG or GC. GG represents the homozygous type in which the nucleotide type at the AX-109906455 locus in the wheat genome is G; the CC genotype indicates the homozygous type in which the nucleotide type at the AX-109906455 locus in the wheat genome is C; the GC genotype represents the heterozygous type in which the nucleotide types at the AX-109906455 locus in the wheat genome are G and C.

[0008] The stripe rust resistance of wheat with the CC genotype at the AX-109906455 locus in the wheat genome is higher than or candidate higher than that of wheat with the GG genotype and / or GC genotype at the AX-109906455 locus in the wheat genome.

[0009] In the present application, the wheat to be tested can be a pure line or an inbred line. The inbred line can be a recombinant inbred line.

[0010] Furthermore, in the method, the substance for detecting the polymorphism or genotype of the AX-109906455 locus can be any of the following A1), A2) or A3):

[0011] A1) A primer composition containing primers for amplifying a wheat genomic DNA fragment including the AX-109906455 locus;

[0012] A2) A PCR reagent containing the primer composition described in A1);

[0013] A3) A kit containing the primer composition described in A1) or the PCR reagent described in A2).

[0014] Furthermore, in the method, the primer composition can be composed of a single-stranded DNA with a nucleotide sequence at positions 22-45 of SEQ ID No.1, a single-stranded DNA with a nucleotide sequence at positions 22-45 of SEQ ID No.2, and a single-stranded DNA with a nucleotide sequence of SEQ ID No.3.

[0015] Furthermore, in the method, the primer composition can be composed of the single-stranded DNA shown in SEQ ID No.1, the single-stranded DNA shown in SEQ ID No.2, and the single-stranded DNA shown in SEQ ID No.3.

[0016] Furthermore, in the method, the method for detecting the genotype of the above-mentioned AX-109906455 locus of the wheat to be tested includes using the genomic DNA of the wheat to be identified as a template, performing PCR amplification using the above-mentioned primer composition to obtain a PCR product; determining the genotype of the AX-109906455 locus according to the sequencing result or fluorescence signal of the PCR product.

[0017] Further, the method specifically includes the following operation steps:

[0018] S1) Extract the genomic DNA of the wheat to be tested;

[0019] S2) Using the genomic DNA extracted in S1) as a template and the above primer composition as amplification primers, perform PCR amplification to obtain a PCR amplification product;

[0020] S3) Judge or assist in judging the stripe rust resistance of the wheat to be tested according to the genotype of the AX-109906455 locus of the PCR amplification product;

[0021] Further, in the above method, in step S3), the genotype of the AX-109906455 locus can be judged according to the sequencing result or fluorescence color development of the PCR product.

[0022] In the present application, the stripe rust resistance of wheat with the CC genotype at the AX-109906455 locus in the wheat genome is higher than or candidate higher than that of wheat with the GG genotype and / or GC genotype at the AX-109906455 locus in the wheat genome.

[0023] Further, the 5' ends of the single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2 are specific fluorescent tag sequences.

[0024] Further, the specific fluorescent tag sequences of the single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2 are different and bind to fluorescent probes of different luminescence types.

[0025] In an embodiment of the present invention, the 1st to 21st nucleotides of the single-stranded DNA shown in SEQ ID No.1 are FAM specific fluorescent tag sequences, and the 1st to 21st nucleotides of the single-stranded DNA shown in SEQ ID No.2 are HEX specific fluorescent tag sequences.

[0026] The CC genotype amplification product binds to FAM fluorescence and shows blue fluorescence; the GG genotype amplification product binds to HEX fluorescence and shows red fluorescence, and the GC genotype amplification product shows green fluorescence. That is, the stripe rust resistance of the wheat to be tested with the amplification product showing blue fluorescence is higher than or candidate higher than that of the wheat to be tested with the amplification product showing red fluorescence or green fluorescence.

[0027] The present application also provides a composition, and the composition contains the substance for detecting the polymorphism or genotype of the AX-109906455 locus.

[0028] The present application also provides the application of the above substance for detecting the polymorphism or genotype of the AX-109906455 locus in the following C1)-C6):

[0029] C1) Identifying or assisting in the identification of wheat stripe rust resistance;

[0030] C2) Preparing a product for identifying or assisting in the identification of wheat stripe rust resistance;

[0031] C3) Screening or assisting in the screening of wheat varieties resistant to stripe rust;

[0032] C4) Preparing a product for screening or assisting in the screening of wheat varieties resistant to stripe rust;

[0033] C5) Wheat breeding and / or assisting breeding;

[0034] C6) Preparing a product for wheat breeding and / or assisting breeding.

[0035] Furthermore, in the above application, the substance for detecting the polymorphism or genotype of the AX-109906455 locus may be any of the following D1), D2), or D3):

[0036] D1) The substance for detecting the polymorphism or genotype of the AX-109906455 locus contains a primer composition for amplifying a wheat genomic DNA fragment including the AX-109906455 locus;

[0037] D2) The substance for detecting the polymorphism or genotype of the AX-109906455 locus is a PCR reagent containing the primer composition;

[0038] D3) A kit containing the primer composition described in D1) or the PCR reagent described in D2).

[0039] Furthermore, in the above composition and / or application, the primer composition is P1 or P2:

[0040] P1. The primer composition is a primer composition composed of a single-stranded DNA with a nucleotide sequence from positions 22 to 45 of SEQ ID No.1, a single-stranded DNA with a nucleotide sequence from positions 22 to 45 of SEQ ID No.2, and a single-stranded DNA with a nucleotide sequence of SEQ ID No.3;

[0041] P2. The primer composition is a primer composition composed of the single-stranded DNA shown in SEQ ID No.1, the single-stranded DNA shown in SEQ ID No.2, and the single-stranded DNA shown in SEQ ID No.3.

[0042] The present application also provides a DNA molecule, and the nucleotide sequence of the DNA molecule is SEQ ID No.4.

[0043] The present application also provides a method for wheat breeding, which includes selecting wheat with the genotype of CC at the above-mentioned AX-109906455 locus as a parent for breeding. The CC genotype represents a homozygous type in which the nucleotide type at the AX-109906455 locus in the wheat genome is C.

[0044] In the present application, the breeding index for the breeding includes wheat stripe rust resistance. The purpose of the breeding includes cultivating wheat with high wheat stripe rust resistance (the wheat stripe rust resistance is higher than that of the parent).

[0045] In the present application, the product can be a reagent or a kit.

[0046] In the present application, the inspection index for the wheat breeding can include wheat stripe rust resistance.

[0047] In the present application, the purpose of the wheat breeding includes cultivating or selecting wheat with stripe rust resistance.

[0048] In the present application, the PCR primers in the primer composition can be labeled with a label or not. The label refers to any atom or molecule that can be used to provide a detectable effect and can be linked to a nucleic acid. Labels include but are not limited to dyes; radioactive labels such as 32P; binding moieties such as biotin; haptens such as digoxin (DIG); luminescent, phosphorescent or fluorescent moieties; and individual fluorescent dyes or fluorescent dyes combined with moieties that can inhibit or shift the emission spectrum through fluorescence resonance energy transfer (FRET). The label can provide a signal detectable by fluorescence, radioactivity, colorimetry, gravimetry, X-ray diffraction or absorption, magnetism, enzyme activity, etc. The label can be a charged moiety (positive charge or negative charge) or, optionally, can be charge-neutral. The label can include a nucleic acid or protein sequence or a combination thereof, as long as the sequence containing the label is detectable. In some embodiments, the nucleic acid is directly detected without a label (e.g., directly reading the sequence).

[0049] Compared with the prior art, the beneficial technical effects achieved by the present application are as follows:

[0050] 1. The present application has first discovered a wheat stripe rust resistance gene QTL and its linked molecular marker. This QTL is named QYR.gaas-2B, located on chromosome 2B, and is tightly linked to AX-109906455 (697.7 Mb).

[0051] 2. The results of detecting the genotypes of the AX-109906455 locus in 111 wheat natural varieties show that the primer combination provided by the present application has good genotyping effect, and this primer can effectively identify whether the genotype of the tested plants based on the AX-109906455 locus is the GG genotype, the CC genotype or the GC genotype.

[0052] 3. Association analysis of the genotypes of natural wheat varieties and the phenotypic resistance to stripe rust indicates a significant correlation between genotypes and phenotypes. This shows that the molecular marker (SNP locus) AX-109906455 and its corresponding primers provided in this application will contribute to the screening of wheat germplasm resources resistant to stripe rust, can be used for molecular marker-assisted selection breeding, and provide material reserves and technical support for the genetic improvement of wheat stripe rust resistance traits. Brief Description of the Drawings

[0053] Figure 1 Genotyping results of 111 wheat varieties by Kasp-2B-YR. Blue represents the Lantian 25 genotype CC, red represents the Huixianhong genotype GG, and green represents the heterozygous GC. Detailed Embodiments

[0054] The following further describes this application in detail in combination with specific embodiments. The provided embodiments are only for clarifying this application, rather than limiting the scope of this application. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and do not limit this application in any way.

[0055] The experimental methods in the following embodiments are all conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0056] The 111 wheat varieties described in the following embodiments are preserved by the applicant and are disclosed in the literature "Genome-Wide Association Mapping of Adult-Plant Resistance to Stripe Rust in Common Wheat (Triticum aestivum). Plant Disease. 2020 Aug; 104(8): 2174-2180. doi: 10.1094 / PDIS-10-19-2116-RE. Epub 2020 May 26." The public can obtain the above biological materials from the applicant, and the obtained biological materials can only be used for verifying the content of this application and cannot be used for other purposes.

[0057] The specific mixed races of stripe rust pathogens described in the following examples are pathogens that are a proportional mixture of CYR30, CYR31, CYR32, CYR33, and CYR34. It is disclosed in the literature "Hu Chaoyue, Wang Fengtao, Lang Xiaowei, et al. Resistance analysis of wheat stripe rust resistance genes to the main prevalent races of Puccinia striiformis f. sp. tritici in China [J]. Scientia Agricultura Sinica, 2022, 55(03): 491-502." The public can obtain the above biological materials from the applicant, and the obtained biological materials can only be used for the verification of the content of this application and cannot be used for other purposes.

[0058] The following examples used GraphPad Prism statistical software to process the data. The experimental results were expressed as mean ± standard deviation, and the t-test was used for testing. *(P < 0.05) indicated significant differences.

[0059] In the quantitative tests in the following examples, unless otherwise specified, three replicates were set, and the results were averaged.

[0060] Example 1. Discovery of a stripe rust resistance gene QTL in wheat Lantian 25 and acquisition of its KASP markers

[0061] I. Obtaining phenotypes

[0062] A recombinant inbred line population consisting of 235 families was constructed using Lantian 25 and Huixianhong. The Lantian 25 / Huixianhong RIL population was planted in Chengdu, Sichuan and Tianshui, Gansu from 2018 - 2019 and 2019 - 2020. A completely randomized block design was adopted, with three replicates, single-row plots, plot length of 2 m, row width of 0.25 m, and 50 seeds evenly sown in each row. Field management was carried out according to local conventions. The RIL population was identified for adult-plant resistance using the current prevalent mixed races of stripe rust in China. The disease index, maximum disease severity (MDS), showed continuous variation in the field, indicating typical quantitative trait inheritance. Using the modified CTAB method (Murray et al., 1980), genomic DNA of young leaves from 235 families was extracted. The DNA concentration was measured using a NanoDrop2000c spectrophotometer, and the DNA samples were adjusted to a standard concentration of 50 ng / μl. Then, the DNA quality was detected using 0.8% agarose gel, and the qualified DNA was subjected to SNP genotyping. SNP analysis was performed using a 50K SNP chip jointly developed by the Institute of Crop Science, Chinese Academy of Agricultural Sciences and Affymetrix Axiom Corporation.

[0063] II. Construction of linkage map

[0064] After removing the markers with heterozygosity between parents and a deletion rate greater than 10%, Icimapping V4.1 was used to remove redundant markers, and then the markers were grouped according to the genetic distance and chromosome position information between the markers. A total of 21 linkage groups were constructed, which corresponded to 21 chromosomes and contained 5941 markers in total.

[0065] III. QTL Analysis

[0066] The IciMapping 4.1 ICIM-ADD method was used for QTL analysis, and the LOD value was selected as 2.5. One stable QTL was mapped on chromosome 2B and named QYR.gaas-2B. It was tightly linked to AX-109906455 (697.7 Mb) and could explain 11.2 - 14.4% of the phenotypic variation under different environmental conditions. Its flanking marker AX-109906455 was transformed into Kasp-2B-YR. The nucleotide sequence of the SNP marker AX-109906455 was the 37th position of SEQ ID No.4, which was a C / G polymorphism.

[0067] For the AX-109906455 locus, primers were designed based on the principle of competitive allele-specific PCR as follows:

[0068] Forward primer AX-109906455-A (SEQ ID No.1): 5'- GAAGGTGACCAAGTTCAT GCT GGGTAGCATTGATCTAAACTCCTG-3' (where the underlined part indicates the FAM fluorescent label sequence);

[0069] Forward primer AX-109906455-B (SEQ ID No.2): 5'- GAAGGTCGGAGTCAACG GATT GGGTAGCATTGATCTAAACTCCTC-3' (where the underlined part indicates the HEX fluorescent label sequence);

[0070] Reverse primer AX-109906455-C (SEQ ID No.3): 5'-CCACCATTCGACCGACAT GA-3'.

[0071] The single-stranded DNA molecules shown in SEQ ID No.1 and SEQ ID No.3 amplified the fragment with the 37th nucleotide of G in SEQ ID No.4, and the fluorescence signal of the fluorescent group bound to the FAM fluorescent label sequence could be read by the instrument;

[0072] The single-stranded DNA molecules shown in SEQ ID No.2 and SEQ ID No.3 are used to amplify the fragment with the 37th nucleotide being C in SEQ ID No.4, and the fluorescence signal of the fluorophore combined with the HEX fluorescence-labeled sequence can be read by an instrument.

[0073] The last nucleotide C / G of the above upstream primer corresponds to the SNP locus C / G at position 109906455 on chromosome 2B, that is, the 37th nucleotide in Sequence 4 of the sequence listing.

[0074] Table 1 KASP primer sequence list for detecting the stripe rust resistance QTL QYR.gaas-2B

[0075]

[0076] Example 2. Genotype and phenotype identification of 111 wheat varieties

[0077] 1. A total of 111 wheat varieties from different sources were planted at the experimental sites in Pixian, Sichuan and Tianshui, Gansu during the years 2014 - 2015 and 2015 - 2016. A completely randomized block design was adopted with three replicates, single-row plots, 2 m in row length, 0.25 m in row width, and 50 seeds were evenly sown in each row. Field management was carried out according to local practices. The adult-plant resistance of the RIL population was identified using the current prevalent mixed races of stripe rust in China, and the maximum disease severity (MDS) was recorded.

[0078] The dominant mixed races of stripe rust were inoculated on 111 wheat plants at the adult-plant stage. When the control variety Huixianhong was severely diseased (disease severity reached 80%), the disease severity of the 111 tested materials was recorded. The disease severity was recorded again every week for a total of three times, and the maximum value among them was taken as the maximum disease severity of the tested materials.

[0079] The MDS value is the percentage of the leaf area covered by pathogen spores in the total leaf area. The statistical method can refer to the literature: Lan Caixia. Mapping of Adult-Plant Resistance QTLs for Stripe Rust and Powdery Mildew in Common Wheat [D]. Chinese Academy of Agricultural Sciences, 2010.

[0080] 2. The genomic DNA of young leaves of 111 families was extracted using the improved CTAB method, and all genomic DNA was detected using the Kasp-2B-YR marker.

[0081] The specific steps are as follows:

[0082] Competitive Allele-Specific PCR: KASP amplification requires three primers, two forward competitive primers (the 5'-ends of the primers have base sequences complementary to the fluorescent groups FAM and HEX, and the other sequences only differ at the SNPs and InDels at the 3'-ends), and one reverse common primer. All reagents used are the supporting reagents provided by LGC company unless otherwise specified. The reagent dosage, usage, and the entire experimental procedure are carried out according to the product manual.

[0083] KASP marker PCR amplification system (4 μL) is as follows: 0.048 μL Primer Mix, 2.0 μL 2×KASP Master Mix, 1.952 μL Template DNA (50 ng / μL). The ratio of Primer Mix is: 12% HEX primer (i.e., the primer shown in SEQ ID No. 2), 12% FAM primer (i.e., the primer shown in SEQ ID No. 1), 30% Common primer (i.e., the primer shown in SEQ ID No. 3, the primer was synthesized by Sangon Biotech, Shanghai). The 2×KASP master Mix contains two universal fluorescent probes and two universal quenching probes synthesized for the upstream primer tag sequences (FAM and HEX).

[0084] Amplification is carried out using a 384-well PCR instrument (BIO-RAD, S1000TM Thermal Cycler), and the program is as follows: 94 °C for 15 min; 94 °C for 20 s, 63 - 55 °C for 1 min (decreasing 1 °C per cycle), 10 cycles; 94 °C for 20 s, 55 °C for 60 s, 32 cycles. The PCR amplification products are placed in an automatic focusing fluorescence multifunctional microplate reader (PHERAstarplus SNP, BMG LABTECH) to read the final fluorescence data, and then the data is imported into the Klustercaller v3.4 software (LGC, Hoddesdon, UK) for genotyping.

[0085] The results are shown in Table 2, Table 3 and Figure 1 <. Among 111 wheat varieties, 77 varieties showed the Lantian 25 genotype CC (fluorescent color was blue), and the average value of stripe rust MDS was 39.1; 31 varieties showed the Huixianhong genotype GG (fluorescent color was red), and the average value of stripe rust MDS was 43.9; 2 varieties showed the heterozygous genotype GC (fluorescent color was green), and the average value of stripe rust MDS was 42.1; statistical tests showed that the gene effect of QYR.caas-2B reached a significant difference (P < 0.05).

[0086] The GG genotype indicates a homozygous type with the nucleotide at locus AX-109906455 in the wheat genome being G; the CC genotype indicates a homozygous type with the nucleotide at locus AX-109906455 in the wheat genome being C; the GC genotype indicates a heterozygous type with the nucleotides at locus AX-109906455 in the wheat genome being G and C.

[0087] The amplification product of the GG genotype binds to the HEX fluorescent group and shows red fluorescence; the amplification product of the CC genotype binds to the FAM fluorescent group and shows blue fluorescence, and the amplification product of the GC genotype shows green fluorescence.

[0088] The results show good genotyping effects, and this primer can effectively identify whether the genotype of the tested plants at locus AX-109906455 is the GG genotype, the CC genotype or the GC genotype.

[0089] The stripe rust resistance of wheat with the CC genotype at locus AX-109906455 in the wheat genome is higher than or potentially higher than that of wheat with the GG genotype and / or GC genotype at locus AX-109906455 in the wheat genome. That is, the stripe rust resistance of the tested wheat with the amplification product showing blue fluorescence is higher than or potentially higher than that of the tested wheat with the amplification product showing red fluorescence or green fluorescence.

[0090] Table 2 Genotype detection results and stripe rust resistance of 111 wheat varieties

[0091]

[0092]

[0093]

[0094]

[0095]

[0096] CC is the genotype of Lantian 25; GG is the genotype of Huixianhong; GC is heterozygous.

[0097] Table 3 Stripe rust resistance effects of 111 natural varieties of QYR.caas-2B

[0098]

[0099] The above has described the present application in detail. For those skilled in the art, without departing from the spirit and scope of the present application and without the need for unnecessary experiments, the present application can be implemented within a relatively wide range under equivalent parameters, concentrations, and conditions. Although specific embodiments of the present application are given, it should be understood that the present application can be further improved. In short, according to the principle of the present application, the present application is intended to include any modifications, uses, or improvements to the present application, including those that depart from the scope disclosed in the present application but are made by using conventional techniques known in the art.

Claims

1. A method for identifying or assisting in identifying wheat stripe rust resistance, characterized in that: Using the wheat genomic DNA to be identified as a template, PCR amplification is performed using the primer combination to obtain a PCR product; the genotype of the AX-109906455 site is determined according to the fluorescent signal of the PCR product; The primer composition consists of the single-stranded DNA shown in SEQ ID No.1, the single-stranded DNA shown in SEQ ID No.2 and the single-stranded DNA shown in SEQ ID No.3; The AX-109906455 site is a SNP site in the wheat genome, and its genotype is CC, GG or GC, wherein GG is a homozygous type in which the nucleotide type of the AX-109906455 site in the wheat genome is G; the CC genotype indicates that the nucleotide type of the AX-109906455 site in the wheat genome is C homozygous; the GC genotype indicates that the nucleotide type of the AX-109906455 site in the wheat genome is a heterozygous type of G and C; The stripe rust resistance of wheat with CC genotype at AX-109906455 in the wheat genome is higher or has a higher potential than that of wheat with GG genotype and / or GC genotype at AX-109906455 in the wheat genome; The CC genotype amplification product combines with FAM fluorescence to display blue fluorescence; the GG genotype amplification product combines with HEX fluorescence to display red fluorescence, and the GC genotype amplification product displays green fluorescence.

2. Use of primer combinations for detecting polymorphism or genotype of AX-109906455 locus in the following C1)-C6): C1) Identify or assist in identifying wheat stripe rust resistance; C2) preparing products for identifying or assisting in identifying resistance to wheat stripe rust; C3) Screening or assisting in the screening of wheat varieties resistant to stripe rust; C4) preparing products for screening or assisting in screening wheat varieties resistant to stripe rust; C5) wheat breeding and / or assisted breeding, wherein the breeding index of the breeding is wheat stripe rust resistance; C6) preparing wheat breeding and / or assisted breeding products, wherein the breeding index is wheat stripe rust resistance; The primer composition consists of the single-stranded DNA shown in SEQ ID No.1, the single-stranded DNA shown in SEQ ID No.2 and the single-stranded DNA shown in SEQ ID No.

3.

3. A method for wheat breeding, characterized in that: The method comprises selecting wheat having a CC genotype at the AX-109906455 locus in claim 1 as a parent for breeding, wherein the CC genotype indicates that the nucleotide type at the AX-109906455 locus in the wheat genome is a homozygous type of C; The breeding index of the breeding is wheat stripe rust resistance.

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