Molecular markers and specific primers associated with wheat storage protein and spike grain number and their application

Abstract

The application discloses a molecular marker and specific primer related to wheat storage protein and a plurality of spike grain number characters and application. The application belongs to the technical field of biotechnology and particularly relates to a molecular marker and specific primer related to wheat storage protein and a plurality of spike grain number characters and application. The application can be applied to identifying or assisting in identifying wheat storage protein content, spike grain number and wheat breeding by detecting the polymorphism or genotype of an InDel molecular marker in a wheat genome or detecting a substance for genotyping. The InDel molecular marker is a DNA molecule with a nucleotide sequence of 197-437 in SEQ ID No. 3 in the sequence listing. The genotype of the InDel molecular marker is identified by using a primer composition composed of SEQ ID No. 1 and SEQ ID No. 2, so that the efficiency of wheat breeding can be improved, and a dominant variety with high wheat storage protein content and spike grain number can be selected.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to molecular markers and specific primers and their applications related to wheat storage proteins and grain number per ear. Background Technology

[0002] Wheat grain storage proteins mainly include glutenin and prolamins. The composition and content of storage proteins determine the rheological properties of wheat dough. Changing the composition and content of grain storage proteins is of great significance for improving the processing quality of wheat flour.

[0003] Using molecular interaction techniques, a transcription factor, TaAUXIN, was identified that binds to the promoter of a storage protein gene. Overexpression of the gene encoding TaAUXIN reduces glutenin content in grains, significantly altering dough strength, elasticity, and extensibility, thus negatively impacting bread processing quality. Furthermore, overexpression reduces grain number per ear, indicating that this gene is a negative regulator of bread processing quality and yield. Identifying allelic variants with low TaAUXIN gene expression could contribute to breeding wheat varieties with strong gluten and high grain number per ear, improving the processing of bread and other products. Summary of the Invention

[0004] The problem to be solved by this invention is how to identify or assist in the identification of wheat storage protein content and the number of grains per ear.

[0005] To address the above technical problems, this invention provides the application of InDel molecular markers or substances for detecting InDel molecular markers, and verifies the practicality of the markers, providing reliable molecular markers for identifying or assisting in the breeding of wheat varieties.

[0006] This invention first provides the application of a substance for detecting polymorphisms or genotypes of the InDel molecular marker in the wheat genome, or a substance for detecting genotypes, in any of the following:

[0007] (1) To identify or assist in the identification of wheat storage protein content and the number of grains per ear;

[0008] (2) Screening or breeding wheat plants, strains, varieties or lines with high wheat storage protein content and a large number of grains per spike.

[0009] (3) Screening or breeding wheat plants, strains, varieties or lines with low wheat storage protein content and few grains per spike.

[0010] (4) Wheat breeding;

[0011] (5) Prepare products for identification or auxiliary identification of wheat storage protein content and number of grains per ear;

[0012] (6) Prepare or select wheat single plants, lines, strains or varieties with high wheat storage protein content and many grains per ear;

[0013] (7) Prepare or select products of wheat single plants, lines, strains or varieties with low wheat storage protein content and few grains per spike.

[0014] (8) Prepare products for wheat breeding.

[0015] The InDel molecular marker is a single-stranded DNA molecule whose nucleotide sequence is positions 197 to 437 of SEQ ID No. 3 in the sequence listing. Using the wheat Chinese Spring IWGSC RefSeq v1.0 (https: / / urgi.versailles.inra.fr / jbrowseiwgsc / gmod_jbrowse / ?data=myData / IWGSC_RefSeq_v1.0) as a reference genome, the InDel molecular marker is a deletion type in Chinese Spring, that is, the InDel molecular marker is deleted at positions 460732450 to 260732451 bp on chromosome 6A of the genome.

[0016] The nucleotide sequence of the other strand of the InDel molecular marker is reverse complementary to positions 197 to 437 of SEQ ID No. 3.

[0017] The InDel molecular marker was named TaAUXIN-STS.

[0018] In this paper, the genotype containing the molecular marker TaAUXIN-STS is defined as the insertion type (abbreviated as the allelic variant InDel-AA genotype). Wheat varieties with the InDel-AA genotype have high storage protein content and high number of grains per ear. The nucleotide sequence of the amplification product of the InDel-AA genotype (which contains a fragment of 850 bp) is SEQ ID No. 3. The genotype not containing the molecular marker TaAUXIN-STS is defined as the deletion type (abbreviated as the allelic variant InDel-BB genotype). Wheat varieties with the InDel-BB genotype have low storage protein content and low number of grains per ear. The nucleotide sequence of the amplification product of the InDel-BB genotype (which contains a fragment of 609 bp) is SEQ ID No. 4.

[0019] The present invention also provides a method for identifying or assisting in the identification of wheat storage protein content and grains per spike, the method comprising detecting the genotype of the InDel molecular marker described above in the wheat to be tested, and identifying or assisting in the identification of wheat storage protein content and grains per spike based on the genotype of the wheat to be tested: the storage protein content and grains per spike of wheat with the InDel-AA genotype are higher than those of wheat with the InDel-BB genotype; the genome of wheat with the InDel-AA genotype contains the InDel molecular marker described above; the genome of wheat with the InDel-BB genotype does not contain the InDel molecular marker described above.

[0020] In one specific embodiment, the method for identifying or assisting in the identification of wheat storage protein content and the number of grains per ear may specifically include the following steps:

[0021] (1) Using the genomic DNA of the wheat variety material to be identified as a template, PCR amplification was performed using the primer pair to obtain the PCR product;

[0022] (2) The wheat storage protein content and the number of grains per spike are identified based on whether the PCR product contains the InDel molecular marker.

[0023] The wheat varieties whose PCR products contain the InDel molecular marker described above have storage protein content and grains per spike that are higher than or greater than those whose PCR products do not contain the InDel molecular marker described above.

[0024] The primers are primer compositions consisting of a forward primer and a reverse primer. The forward primer is a single-stranded DNA that specifically binds upstream of the InDel molecular marker in wheat genomic DNA, and the reverse primer is a single-stranded DNA that specifically binds downstream of the InDel molecular marker in wheat genomic DNA.

[0025] In one specific embodiment, the primer composition may be STS-F and STS-R.

[0026] The STS-F:5'-CCTTCGGCTCACTCTAGTGCATG-3' (SEQ ID No. 1);

[0027] The STS-R:5'-CTGCGCTCGCGTGTCTGAG-3' (SEQ ID No. 2).

[0028] This invention also provides the application of the methods described above in wheat breeding.

[0029] The present invention also provides a method for wheat breeding, comprising detecting the type of genotype of the InDel molecular marker described above in the wheat genome, selecting wheat with genotype InDel-AA as a parent for breeding, wherein the genotype InDel-AA is a gene type containing the InDel molecular marker described above.

[0030] The present invention also provides a product for detecting the polymorphism or genotype of the molecular markers described above, the product containing the substances described above, and the product may be any one of them:

[0031] C1) Products that detect nucleotide polymorphisms or genotypes related to wheat storage protein content and the number of grains per ear;

[0032] C2) Products used for identifying or assisting in the identification of wheat storage protein content and the number of grains per ear;

[0033] C3) Products used in wheat breeding;

[0034] C4) Screening or breeding of wheat single plants, lines, strains or varieties with high wheat storage protein content and high number of grains per spike;

[0035] C5) Screening or breeding of wheat plants, strains, varieties or products with low wheat storage protein content and low number of grains per spike.

[0036] In the above applications or products, the substance may be D1), D2), or D3):

[0037] D1) The substance described is a primer composition for amplifying wheat genomic DNA fragments including the InDel described above;

[0038] D2) The substance described is a PCR reagent containing the primer composition described in D1;

[0039] D3) The substance is a kit containing the primer composition described in D1) or the PCR reagent described in D2).

[0040] In the above applications or products, the primer composition for amplifying wheat genomic DNA fragments, including the InDel molecular marker described in D1), consists of SEQ ID No. 1 and SEQ ID No. 2.

[0041] In one specific embodiment, the primer composition may be STS-F and STS-R.

[0042] The STS-F:5'-CCTTCGGCTCACTCTAGTGCATG-3' (SEQ ID No. 1);

[0043] The STS-R:5'-CTGCGCTCGCGTGTCTGAG-3' (SEQ ID No. 2).

[0044] The present invention also provides a DNA molecule whose nucleotide sequence is positions 197 to 437 of SEQ ID No. 3 in the sequence listing.

[0045] The present invention also provides the use of the above-described DNA molecule in any of the following:

[0046] (1) To identify or assist in the identification of wheat storage protein content and the number of grains per ear;

[0047] (2) Screening or breeding wheat plants, strains, varieties or lines with high wheat storage protein content and a large number of grains per spike.

[0048] (3) Screening or breeding wheat plants, strains, varieties or lines with low wheat storage protein content and few grains per spike.

[0049] (4) Wheat breeding;

[0050] (5) Prepare products for identification or auxiliary identification of wheat storage protein content and number of grains per ear;

[0051] (6) Prepare or select wheat single plants, lines, strains or varieties with high wheat storage protein content and many grains per ear;

[0052] (7) Prepare or select products of wheat single plants, lines, strains or varieties with low wheat storage protein content and few grains per spike.

[0053] (8) Prepare products for wheat breeding.

[0054] Genetic variations of TaAUXIN were obtained through the wheat omics website (http: / / 202.194.139.32 / getfasta / index.html) and PCR amplification of the TaAUXIN promoter. An insertion or deletion of 241 nucleotides at position 419 upstream of the start codon of the TaAUXIN gene resulted in two genotypes: InDel-AA and InDel-BB. A sequence tagged site (STS) marker was developed based on the 241 nucleotide insertion / deletion (InDel) in the TaAUXIN promoter region to effectively distinguish between these two genotypes. This molecular marker was used to genotype a population comprising 156 wheat lines from the Huang-Huai wheat region of my country. Genetic effect analysis revealed that wheat varieties with the InDel-AA genotype had higher grain storage protein content and higher grains per spike, providing a usable marker for marker-assisted selection in wheat breeding.

[0055] Compared to the currently popular high-throughput genotyping technologies based on sequencing and microarrays, the operation process and equipment requirements for genotyping using this marker are relatively simple. The products are amplified by a PCR instrument, and genotyping can be achieved using an agarose gel electrophoresis device and a UV lamp. The detection cost is very low, and the scale of the test samples can be flexibly adjusted. Attached Figure Description

[0056] Figure 1 Genotyping of 1-6 genes in 156 representative wheat lines from the Huang-Huai wheat region was performed using the molecular marker TaAUXIN-STS. M: DNA marker; 1: Yannong 15; 2: Gan 6172; 3: Zhongmai 895; 4: Aikang 58; 5: Xiaoyan 6; 6: Zhong 892

[0057] Figure 2 The effect of TaAUXIN allelic variation on storage protein and grain number per ear. Significant difference P < 0.05 (*) Detailed Implementation

[0058] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0059] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0060] Unless otherwise specified, all quantitative experiments in the following examples are performed in triplicate.

[0061] The 156 natural populations of wheat from the Huang-Huai region in the following examples have been documented in: Li F, Wen W, Liu J, Zhang Y, Cao S, He Z, Rasheed A, Jin H, Zhang C, Yan J, Zhang P, Wan Y, Xia X (2019) Genetic architecture of grain yield in bread wheat based on genome-wide association studies. BMC Plant Biology 19(1):168). This biological material is available to the public from the applicant and is intended solely for the replication of experiments of this invention and may not be used for any other purpose.

[0062] The wheat variety Aikang 58 used in the following examples has been described in: Li F, Wen W, Liu J, Zhang Y, Cao S, He Z, Rasheed A, Jin H, Zhang C, Yan J, Zhang P, Wan Y, Xia X (2019) Genetic architecture of grain yield in bread wheat based on genome-wide associations studies. BMC Plant Biology 19(1):168). This biological material is available to the public from the applicant and is intended solely for the purpose of replicating experiments of this invention; it may not be used for any other purpose.

[0063] The wheat variety Yannong 15 used in the following examples has been described in: Li F, Wen W, Liu J, Zhang Y, Cao S, He Z, Rasheed A, Jin H, Zhang C, Yan J, Zhang P, Wan Y, Xia X (2019) Genetic architecture of grain yield in bread wheat based on genome-wide associations studies. BMC Plant Biology 19(1):168). This biological material is available to the public from the applicant and is intended solely for the purpose of replicating experiments of this invention and may not be used for any other purpose.

[0064] The following examples use EXCEL statistical software to process the data. The experimental results are expressed as mean ± standard deviation. The two-sample equal variance hypothesis test is used, and P < 0.05 (*) indicates that there is a significant difference.

[0065] Example 1: Discovery of TaAUXIN-STS Molecular Markers

[0066] 1. Identification of allelic variations in TaAUXIN

[0067] The wheat genome ID (TraesCS6D02G378300) of the transcription factor encoding gene TaAUXIN was entered into the wheat genome variation database (Wheat-SnpHub-Portal http: / / wheat.cau.edu.cn / Wheat_SnpHub_Portal / ). A genome search of 80 varieties (CC group) in this database revealed a 241-nucleotide insertion / deletion variation at nucleotide 419 upstream of the start codon of the TaAUXIN gene.

[0068] Using genomic DNA from wheat varieties Aikang 58 and Yannong 15 as templates, PCR amplification and first-generation sequencing identified a 241-nucleotide fragment at position 419 upstream of the start codon of the TaAUXIN gene as either an insertion or deletion. Aikang 58 did not have this 241-nucleotide insertion at position 419, and its genotype was defined as InDel-BB. Aikang 58 had a storage protein content of 15.25%. Yannong 15, on the other hand, had a 241-nucleotide insertion at position 419, and its genotype was defined as InDel-AA. Yannong 15 had a storage protein content of 16.35%, higher than that of Aikang 58. The formula for calculating wheat storage protein content is: Protein content (14% wet basis) = Protein (dry basis) content × (1-14%).

[0069] Using the wheat Chinese Spring IWGSC Refseq v1.0 (https: / / urgi.versailles.inra.fr / jbrowseiwgsc / gmod_jbrowse / ?data=myData / IWGSC_RefSeq_v1.0) as the reference genome, a 241-nucleotide segment located at position 419 upstream of the start codon of the TaAUXIN gene is deleted in the Chinese Spring genome, specifically between 460732450 and 260732451 bp on chromosome 6A. Since variations in the TaAUXIN gene promoter region may be crucial for regulating gene expression, a molecular marker was developed based on this allelic variant site for genotyping. This molecular marker at this allelic variant site is named the molecular marker TaAUXIN-STS.

[0070] 2. Development of the molecular marker TaAUXIN-STS and its specific primer set

[0071] Given that variations in the promoter region of the TaAUXIN gene may be crucial for regulating its expression, identification primers for the molecular marker TaAUXIN-STS were developed based on differences in sequence length.

[0072] Using genomic DNA from wheat varieties Aikang 58 and Yannong 15 as templates, PCR amplification was performed using primer sets. The specific steps are as follows:

[0073] 1) From the wheat omics website ( http: / / 202.194.139.32 / getfasta / index.html Obtain the sequence 1 kb upstream of the start codon of the TaAUXIN gene (Sequence 3). Compare this DNA segment with sequences that are homologous or highly similar to this region on the GSP website. http: / / probes.pw.usda.gov / GSP / index.php ).

[0074] 2) Based on the alignment results, determine the specific sites of the target DNA segment and design a specific primer set accordingly:

[0075] STS-F:5'-CCTTCGGCTCACTCTAGTGCATG-3' (SEQ ID No. 1);

[0076] STS-R: 5'-CTGCGCTCGCGTGTCTGAG-3' (SEQ ID No. 2).

[0077] The PCR amplification system for the molecular marker TaAUXIN-STS (20 μl) included 2 μl of genomic DNA (approximately 100 ng / μl) as a template, 10 μl of 2×PCR mix (PowerPol 2×PCR Mix with Dye, ABclonal), 2 μl each of forward and reverse primers STS-F / STS-R (primer concentration 10 μM), and 4 μl of deionized water. PCR was performed in a 96-well plate.

[0078] PCR amplification program: 94℃ for 15 min; 35 cycles: 94℃ for 20 s, 61℃ for 30 s, 72℃ for 30 s; 72℃ for 5 min; store at 16℃.

[0079] The size of PCR amplification products was detected by 1% agarose gel electrophoresis.

[0080] The results showed that PCR amplification using primers STS-F / STS-R yielded a 609 bp fragment in the genomic DNA amplification product of wheat variety Aikang 58 and an 850 bp fragment in the genomic DNA amplification product of wheat variety Yannong 15. Sequencing alignment revealed a 241 nucleotide insertion in the PCR product of wheat variety Yannong 15. The amplified fragments were consistent with the expected size, confirming the effectiveness of this molecular marker in differentiating the content of wheat storage proteins.

[0081] The genotyping criteria for the wheat molecular marker TaAUXIN-STS are as follows: If the PCR amplification product band size is 609 bp, the wheat genotype is InDel-BB, indicating a germplasm resource with low storage protein content and low grain number per spike; if the PCR amplification product band size is 850 bp, the wheat genotype is InDel-AA, indicating a germplasm resource with high storage protein content and high grain number per spike. Wheat with the InDel-AA genotype exhibits superior storage protein content and / or grain number per spike compared to wheat with the InDel-BB genotype.

[0082] The genotype containing the molecular marker TaAUXIN-STS is defined as the insertion type (abbreviated as the InDel-AA genotype). Wheat varieties with the InDel-AA genotype have high storage protein content and high grain number per ear. The nucleotide sequence of the amplification product of the InDel-AA genotype (which contains an 850bp fragment) is SEQ ID No. 3. The genotype not containing the molecular marker TaAUXIN-STS is defined as the deletion type (abbreviated as the InDel-BB genotype). Wheat varieties with the InDel-BB genotype have low storage protein content and low grain number per ear. The nucleotide sequence of the amplification product of the InDel-BB genotype (which contains a 609bp fragment) is SEQ ID No. 4.

[0083] Example 2: Application of the molecular marker TaAUXIN-STS and its specific primer set in identifying the storage protein content and grain number traits of wheat varieties.

[0084] 1. Field phenotypic identification and data analysis of 156 natural wheat populations in the Huang-Huai region

[0085] 156 natural wheat varieties from the Huang-Huai wheat region were planted in Anyang, Henan and Suixi, Anhui in 2012-2013 and 2013-2014, and in Anyang, Henan and Gaoyi, Hebei in 2014-2015. A completely randomized block design with three replicates was used, with single-row plots, row length 1.5m, row width 0.2m, and 50 grains / row. Field management practices followed local wheat field management standards.

[0086] Protein content was analyzed using a Perten DA 7200 instrument (Perten, Springfield, IL, USA) near-infrared reflectance spectrometer. The average phenotypic data for multi-environment storage proteins and the number of grains per ear are shown in Table 1.

[0087] 2. Analysis of genotypes and genetic effects of wheat varieties in the Huang-Huai wheat region using the molecular marker TaAUXIN-STS.

[0088] Genomic DNA was extracted from 156 wheat varieties from the Huang-Huai wheat region as templates. Using the identification primer set STS-F / STS-R of the molecular marker TaAUXIN-STS, PCR amplification was performed according to the PCR system and procedure in step 2 of Example 1 to obtain PCR amplification products.

[0089] The genotype of the molecular marker TaAUXIN-STS was determined by electrophoresis using 1% agarose gel (Table 1). Figure 1 ).

[0090] The genotyping results and phenotypic data were analyzed using a t-test in Excel to determine the genetic effects of TaAUXIN on storage protein and ear grain number weight. The genetic analysis results for storage protein and ear grain number are as follows: Figure 2 As shown. Figure 2 Statistical analysis of the two genotypes, InDel-BB and InDel-AA, on seed protein content and thousand kernel weight. * indicates significant difference (P<0.05).

[0091] Table 1. Genotyping and phenotypic data of the molecular marker TaAUXIN-STS on 156 representative wheat lines from the Huang-Huai wheat region.

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098] Table 2. Analysis of the genotype relationship between TaAUXIN-STS and wheat storage protein content phenotype

[0099]

[0100] Table 3. Statistical association analysis between TaAUXIN-STS genotype and grain number phenotype

[0101]

[0102]

[0103] Note: The statistical analysis uses the two-sample equal variance hypothesis test method.

[0104] As shown in Tables 1, 2, and 3, the storage protein content (mean 15.75% ± 1.03%) and / or number of grains per spike (mean 41.43 ± 6.95) of wheat with the InDel-AA genotype are superior to those of wheat with the InDel-BB genotype (mean 15.17% ± 1.09%) and / or number of grains per spike (mean 38.03 ± 5.40). In other words, the storage protein content and number of grains per spike of TaAUXIN-STS wheat with the InDel-AA genotype are higher than those of the InDel-BB homozygous wheat.

[0105] In the InDel-AA homozygous wheat genome, there is a 241-nucleotide insertion at position 419 upstream of the start codon in the TaAUXIN gene on both chromosomes (wheat genotype is the allelic InDel-AA). This type of germplasm resource is a high-quality germplasm resource with high storage protein content and a large number of grains per spike. In contrast, in the InDel-BB homozygous wheat genome, there is no 241-nucleotide insertion at position 419 upstream of the start codon in the TaAUXIN gene on both chromosomes (wheat genotype is the allelic InDel-BB). This type of germplasm resource is a low-quality germplasm resource with low storage protein content and a small number of grains per spike. When breeding high-quality germplasm resources with high storage protein content and a large number of grains per spike, it is best to select wheat with the InDel-AA genotype (TaAUXIN-STS molecular marker), i.e., wheat with the InDel-AA homozygous genotype, as the parent for breeding.

[0106] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. Application of substances used to detect InDel molecular marker polymorphisms or genotypes in the wheat genome in any of the following: (1) To identify or assist in the identification of wheat storage protein content and the number of grains per ear; (2) Screening or breeding wheat plants, strains, varieties or lines with high storage protein content and many grains per spike; (3) Screening or breeding wheat plants, strains, varieties or lines with low storage protein content and few grains per spike; (4) Prepare products for identification or auxiliary identification of wheat storage protein content and number of grains per ear; (5) To prepare or select products of wheat single plants, lines, strains or varieties with high wheat storage protein content and many grains per ear; (6) To prepare or select products of wheat single plants, lines, strains or varieties with low wheat storage protein content and few grains per ear; The InDel molecular marker is a single-stranded nucleotide sequence that is the DNA molecule at positions 197 to 437 of SEQ ID No. 3 in the sequence listing; The storage protein content and number of grains per spike of wheat with genotype InDel-AA are higher than those of wheat with genotype InDel-BB; the genome of wheat with genotype InDel-AA contains the InDel molecular marker; the genome of wheat with genotype InDel-BB does not contain the InDel molecular marker.

2. The application according to claim 1, characterized in that: The substance is either D1), D2), or D3). D1) The substance is a primer composition for amplifying wheat genomic DNA fragments including the InDel mentioned above; D2) The substance described is a PCR reagent containing the primer composition described in D1; D3) The substance is a kit containing the primer composition described in D1) or the PCR reagent described in D2).

3. The application according to claim 2, characterized in that: The primer composition described in D1) consists of SEQ ID No. 1 and SEQ ID No.

2.

4. A method for identifying or assisting in the identification of wheat storage protein content and grains per spike, the method comprising detecting the genotype of the InDel molecular marker of claim 1 in the wheat to be tested, and identifying or assisting in the identification of wheat storage protein content and grains per spike based on the genotype of the InDel molecular marker of the wheat to be tested: the storage protein content and grains per spike of wheat with the InDel-AA genotype are higher than those of wheat with the InDel-BB genotype; the genome of wheat with the InDel-AA genotype contains the InDel molecular marker of claim 1; the genome of wheat with the InDel-BB genotype does not contain the InDel molecular marker of claim 1.

5. The application of the method of claim 4 in breeding wheat storage protein content and grains per ear.

6. A method for breeding wheat based on storage protein content and number of grains per spike, characterized by: The method includes detecting the genotype of the InDel molecular marker of claim 1 in the wheat genome, selecting wheat with genotype InDel-AA as a parent for breeding, wherein the genotype InDel-AA is a gene type containing the InDel molecular marker of claim 1.

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