Molecular marker for identifying activity of superoxide dismutase in wheat grains and application of molecular marker

Abstract

The invention discloses a molecular marker for identifying the activity of superoxide dismutase in wheat grains and application of the molecular marker. The invention provides a complete set of primers for identifying the content of superoxide dismutase in wheat grains. The complete set of primers consists of a primer pair 1 and a primer pair 2, the primer pair 1 is composed of single-stranded DNA (deoxyribonucleic acid) as shown in SEQ ID No. 1 and SEQ ID No. 2; the primer pair 2 is composed of single-stranded DNA (deoxyribonucleic acid) as shown in SEQ ID No. 3 and SEQ ID No. 4. The molecular marker can rapidly identify the allelic variation of TaSOD-B2a and TaSOD-B2b and the correlation between the allelic variation of TaSOD-B2a and TaSOD-B2b and the SOD content of wheat, and can screen out wheat varieties with high SOD content, so that the breeding pace of new high-quality wheat varieties is accelerated, and the molecular marker has important theoretical significance and economic value for auxiliary selection of the wheat varieties with high SOD content by using the molecular marker.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular marker for identifying superoxide dismutase activity of wheat grains and an application thereof. Background Art

[0002] The color of flour and its products is an important part of genetic improvement of wheat quality, and the color of flour has become an important factor restricting the quality of flour and its products. Superoxide dismutase activity is the main factor affecting the color and processing quality of flour products, causing flour and flour products to lose color and turn white during the processing and storage of wheat products. Superoxide dismutase can oxidize the conjugated double bonds of pigment molecules such as carotenoids in flour and flour products into single bonds through the molecular oxygen released during the dismutation reaction, thereby bleaching the color of flour. The rheological properties of dough are one of the important factors affecting wheat quality. Superoxide dismutase affects the intramolecular or intermolecular sulfhydryl and disulfide bond exchange reactions of proteins in the dough to produce more disulfide bonds, bring together the amino acid residues on the peptide chain, and form a network structure in which the arrangement of protein molecules is more orderly, thereby stabilizing the conformation of the protein and improving the rheological properties of the dough. In addition, superoxide dismutase can also dismutate superoxide anion free radicals (O 2- ), the generated hydrogen peroxide and molecular oxygen further degrade the dietary fiber components in the flour into components such as oligosaccharides with prebiotic effects.

[0003] Wheat superoxide dismutase, also known as SOD enzyme, plays an important role in the flour processing. This enzyme can inhibit superoxide anions in flour and prevent them from damaging the plant pigments in flour. It is precisely because of the presence of SOD enzyme that the flour we see can be fuller and brighter in color.

[0004] Not only that, SOD enzyme can also protect important nutrients in flour and maintain its good form. During the storage and use of flour, due to exposure to air, some adverse factors will be generated in the flour, such as oxidation reaction, moisture, etc. These factors can easily reduce the content and stability of nutrients in flour. However, SOD enzyme, as an effective antioxidant, can degrade these harmful substances and maintain the integrity of important components in flour.

[0005] The importance of wheat superoxide dismutase in flour processing is not only reflected in the color of flour. It can also improve the quality and taste of flour, making the prepared pasta more delicious. Therefore, improving superoxide dismutase activity through genetic and conventional breeding methods, and using molecular marker-assisted selection to quickly improve the color of wheat flour products and increase the added value of wheat have important theoretical significance and economic value. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a set of primers for identifying superoxide dismutase activity of wheat grains and / or how to identify superoxide dismutase activity of wheat grains.

[0007] In order to solve the above technical problems, the present invention first provides a primer combination, which can be composed of a primer pair 1 and a primer pair 2; the primer pair 1 is composed of a single-stranded DNA shown in SEQ ID No.1 and a single-stranded DNA shown in SEQ ID No.2; the primer pair 2 is composed of a single-stranded DNA shown in SEQ ID No.3 and a single-stranded DNA shown in SEQ ID No.4.

[0008] In order to solve the above technical problems, the present invention also provides a reagent and / or a kit for identifying superoxide dismutase activity in plant seeds, wherein the reagent and / or the kit may contain the primer combination described above.

[0009] In the above reagents and / or kits, the final concentration of each primer in the primer pair 1 and / or the primer pair 2 in the PCR reaction system can be 10 -7 mol / L.

[0010] In order to solve the above technical problems, the present invention also provides any of the following applications of the primer composition and / or the reagent and / or kit described above: P1. Application in identification or auxiliary identification of SOD activity in wheat to be tested; P2. Application in detecting or assisting in detecting wheat SOD activity; P3. Application in the preparation and identification of products or in assisting the identification of wheat SOD activity to be tested; P4. Application in the preparation of products for detecting or assisting in detecting the activity of wheat SOD to be tested; P5. Application in the preparation of products for detecting or assisting in detecting the TaSOD-B2 genotype of wheat to be detected; P6. Application in breeding or improvement of wheat SOD activity phenotype.

[0011] In order to solve the above technical problems, the present invention also provides the use of the primer composition described above or the reagent and / or kit described above in detecting or assisting in detecting whether the wheat to be tested is high SOD activity wheat or low SOD activity wheat; The SOD activity in the low SOD activity wheat grains may be less than or equal to 1746.27 U / g; The SOD activity in the wheat grains with high SOD activity may be greater than or equal to 1828.67 U / g.

[0012] In order to solve the above technical problems, the present invention also provides a method for detecting or assisting in detecting the SOD activity of wheat to be tested, and the method may include the following steps: The genomic DNA of the wheat to be tested is subjected to PCR amplification using the primer combination and / or the reagent and / or the kit described above to obtain a PCR amplification product; and the SOD activity of the wheat to be tested is detected based on the size of the PCR amplification product.

[0013] In the above method, the wheat to be tested, whose PCR amplification product of the primer pair 1 contains a DNA fragment of 643 bp in size, and whose PCR amplification product of the primer pair 2 does not contain a DNA fragment of 1106 bp in size, is named as Group A wheat material; the wheat plant to be tested, whose PCR amplification product of the primer pair 2 contains a DNA fragment of 1106 bp in size, and whose PCR amplification product of the primer pair 1 does not contain a DNA fragment of 643 bp in size, is named as Group B plant material, and the SOD activity of the grains of the Group A wheat material is lower than the SOD activity of the grains of the Group B wheat material.

[0014] The SOD activity of the wheat material of group A may be less than or equal to 1746.27 U / g; the wheat material of group A may be wheat with low SOD activity; The SOD activity of the Group B wheat material may be greater than or equal to 1828.67 U / g; and the Group B wheat material may be high SOD activity wheat.

[0015] One strand of the 643 bp DNA fragment may correspond to nucleotides 2689-3331 of SEQ ID No. 5 in the sequence listing; one strand of the 1106 bp DNA fragment may correspond to nucleotides 2689-3794 of SEQ ID No. 6 in the sequence listing.

[0016] In order to solve the above technical problems, the present invention also provides a method for detecting or assisting in detecting that the wheat to be tested contains TaSOD-B2a haplotype and / or TaSOD-B2b haplotype, comprising the following steps: Using the primer pair 1 and the primer pair 2 described above to perform PCR amplification on the genomic DNA of the wheat to be tested, respectively, to obtain PCR amplification products; according to the length of the DNA fragment of the PCR amplification product band, confirming whether the wheat to be tested contains TaSOD-B2a haplotype and / or TaSOD-B2b haplotype; The PCR amplification product may contain a DNA fragment of 643 bp in length, and the wheat to be tested may contain the TaSOD-B2a haplotype; the PCR amplification product may contain a DNA fragment of 1106 bp in length, and the wheat to be tested may contain the TaSOD-B2b haplotype; The TaSOD-B2a haplotype may correspond to nucleotides 2689-3331 of SEQ ID No. 5 in the sequence listing, and the TaSOD-B2b haplotype may correspond to nucleotides 2689-3794 of SEQ ID No. 6 in the sequence listing.

[0017] In order to solve the above technical problems, the present invention also provides a method for cultivating wheat with low SOD activity, comprising the following steps: breeding with plant materials of group A as parents to cultivate wheat with low SOD activity; The plant material of group A can meet the following conditions: the amplification product obtained by PCR amplification of genomic DNA using the primer pair 1 described above contains a DNA fragment of 643 bp in size, and the amplification product obtained by PCR amplification of genomic DNA using the primer pair 2 described above does not contain a DNA fragment of 1106 bp in size; one chain of the DNA fragment of 643 bp in size can correspond to nucleotides 2689-3331 of SEQ ID No.5 in the sequence listing; one chain of the DNA fragment of 1106 bp in size can correspond to nucleotides 2689-3794 of SEQ ID No.6 in the sequence listing; the SOD activity in the wheat grains with low SOD activity is less than or equal to 1746.27 U / g.

[0018] In order to solve the above technical problems, the present invention also provides a method for cultivating wheat with high SOD activity, comprising the following steps: using group B wheat as a parent to carry out wheat breeding to cultivate wheat with high SOD activity; The group B wheat can meet the following conditions: the amplification product obtained by PCR amplification of genomic DNA using the primer pair 2 mentioned above contains a fragment with a size of 1106 bp, and the amplification product obtained by PCR amplification of genomic DNA using the primer pair 1 mentioned above does not contain a DNA fragment with a size of 643 bp; one chain of the DNA fragment with a size of 643 bp can correspond to nucleotides 2689-3331 of SEQ ID No.5 in the sequence listing; one chain of the DNA fragment with a size of 1106 bp can correspond to nucleotides 2689-3794 of SEQ ID No.6 in the sequence listing; the SOD activity in the grains of the wheat with high SOD activity is greater than or equal to 1828.67 U / g.

[0019] In the above, the wheat to be tested may specifically be at least one of the wheat listed in Table 1.

[0020] Experiments have shown that the molecular markers of the present invention can quickly identify the allelic variation of TaSOD-B2a and TaSOD-B2b and the level of their SOD activity. The molecular markers used in the present invention can quickly screen out wheat varieties (materials) with higher SOD activity, which not only accelerates the breeding of high-quality new wheat varieties, but also has important theoretical significance and economic value for the use of molecular markers to assist in the selection of wheat varieties with high SOD activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The electrophoresis test diagram of some samples. M is 2000 markers, and 01-10 represent different material samples. DETAILED DESCRIPTION

[0022] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0023] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0024] Example 1: Method for Identifying Superoxide Dismutase (SOD) Activity in Wheat Grains 1. Identification of molecular markers for superoxide dismutase activity in wheat grains 1.1 SOD activity-related alleles There are two allelic variation forms of the TaSOD-B2 gene in wheat: one is shown in the nucleotide molecule from 2689 to 3331 from the 5' end in SEQ ID No. 5 of the sequence listing (named as the allele TaSOD-B2a haplotype); the other is shown in the nucleotide molecule from 2689 to 3794 from the 5' end in SEQ ID No. 6 of the sequence listing (named as the TaSOD-B2b haplotype).

[0025] 1.2 Primer design Based on the two allelic variant forms of the TaSOD-B2 gene, specific primer pair 1 and primer pair 2 were designed as follows: Upstream primer of primer pair 1 (SEQ ID No. 1): 5′-ACATGTTTGTTATGCAGTGGCTA-3′; Downstream primer of primer pair 1 (SEQ ID No. 2): 5′-TAGGTGCAGAATGCAAGTCAATA-3′; Upstream primer of primer pair 2 (SEQ ID No. 3): 5′-ACATGTTTGTTATGCAGTGGCTT-3′; Downstream primer of primer pair 2 (SEQ ID No. 4): 5'-CGAATGTGATTGTACGTATGAC-3'.

[0026] 2. Method for Identifying Superoxide Dismutase Activity in Wheat Grains 2.1 Extract genomic DNA from wheat grains to be tested; 2.2 PCR amplification Using the genomic DNA obtained above as a template, PCR amplification was performed using primer pair 1 and primer pair 2 in step 1.2, respectively, to obtain two PCR amplification products; The reaction system for PCR amplification: template DNA about 80ng, Taq DNA Polymerase (TIANGEN, code: ET101) 0.25µl, 10×Taq BufferⅠ 2.5µl (TIANGEN, code: ET101), dNTP Mixture (TIANGEN, code: CD111, 2.5 mM each) 1µl, upstream primer and downstream primer 1µl each, and add sterile ultrapure water to make up the reaction system to 25μl.

[0027] The reaction procedure of PCR amplification corresponding to primer pair 1 was as follows: 94°C pre-denaturation for 5 min; 94°C denaturation for 30 s, 54.5°C annealing for 30 s, 72°C extension for 45 s, for a total of 35 cycles; and finally 72°C extension for 8 min; and 4°C insulation. The obtained PCR amplification products were sequenced respectively.

[0028] The reaction procedure of PCR amplification corresponding to primer pair 2 was: 94°C pre-denaturation for 5 min; 94°C denaturation for 30 s, 56°C annealing for 30 s, 72°C extension for 1 min 10 s, for a total of 35 cycles; finally 72°C extension for 8 min; 4°C incubation. The obtained PCR amplification products were sequenced respectively.

[0029] The two PCR amplification products were electrophoresed on 2.0% agarose gel and 1×TAE electrophoresis buffer and then observed under a gel imager.

[0030] 2.3 Definition of superoxide dismutase (SOD) activity in wheat grains: When the SOD activity is less than or equal to 1746.27U / g, it indicates that the wheat grain is low SOD activity wheat; When the SOD activity is greater than or equal to 1828.67U / g, it indicates that the wheat grain is high SOD activity wheat.

[0031] (1) If the PCR amplification product of the primer pair 1 contains a DNA fragment of 643 bp in size (one strand of the DNA fragment corresponds to nucleotides 2689-3331 of SEQ ID No. 5 in the sequence list), and the PCR amplification product of the primer pair 2 does not contain a DNA fragment of 1106 bp in size (one strand of the DNA fragment corresponds to nucleotides 2689-3794 of SEQ ID No. 6 in the sequence list), then the wheat to be tested is low SOD activity wheat; If the PCR amplification product of the primer pair 2 contains a fragment of 1106 bp in size, and the PCR amplification product of the primer pair 1 does not contain a fragment of 643 bp in size, the wheat to be tested is high SOD activity wheat; (2) If the PCR amplification product of the primer pair 1 contains a fragment of 643 bp in size, and the PCR amplification product of the primer pair 2 does not contain a fragment of 1106 bp in size, then the wheat to be tested contains the allele TaSOD-B2a; If the PCR amplification product of the primer pair 2 contains a fragment of 1106 bp in size, and the PCR amplification product of the primer pair 1 does not contain a fragment of 643 bp in size, the wheat to be tested contains the allele TaSOD-B2b.

[0032] The sequence of SEQ ID No.5 is as follows (5'-3'):

[0033] The sequence of SEQ ID No.6 is as follows (5'-3'):

[0034] Example 2: Validation of SOD activity detection molecular markers Experimental materials: 125 main varieties of winter wheat (as shown in Table 1), which are available to the public from the National Germplasm Resource Bank of the Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, and can only be used to replicate the present invention and cannot be used for other purposes.

[0035] The method for identifying superoxide dismutase activity in wheat grains described in Example 1 was used to verify 125 main winter wheat varieties. At the same time, each wheat variety in Table 1 was planted in Anyang, Henan Province. After harvest, the SOD activity of each wheat material grain was detected. The SOD activity of each wheat material was tested twice. If the error of the two test results exceeded 10%, a third repeated test was required, and the results were averaged.

[0036] 1. The experimental method for determining the SOD activity of grains is as follows: Extraction of SOD from common wheat grains: (1) 10 g of wheat seeds were selected and ground into whole wheat flour using a grinder (Laboratory Mill 120, Sweden). 0.1 g of the whole wheat flour was weighed and placed in a 2 mL centrifuge tube.

[0037] (2) Add 1 mL of 0.05 mol·L precooled at 4°C -1 Phosphate buffer (pH 7.8) was placed on a vortexer for oscillation to allow the buffer to fully contact the whole wheat flour, and then the centrifuge tube containing the sample was shaken in an ice bath for 2 hours.

[0038] (3) Centrifuge at 10,000 rpm at 4°C for 15 min.

[0039] (4) Aspirate the supernatant to obtain the crude superoxide dismutase extract, and immediately store it in a 4°C refrigerator for later use.

[0040] 2. Determination of SOD activity by nitroblue tetrazolium (NBT) photoreduction method The present invention uses the nitro blue tetrazolium (NBT) photochemical reduction method to determine the SOD activity. In the presence of oxidizing substances, riboflavin can be reduced by light. The reduced riboflavin is very easy to be reoxidized under aerobic conditions to produce O2, which can reduce nitro blue tetrazolium to blue methylhydrazone, which is in the presence of OD. 560nmThere is maximum absorption at . SOD can remove O2 and thus inhibit the formation of methyl hydrazone. Therefore, after the photoreduction reaction, the deeper the blue color of the reaction solution, the lower the enzyme activity, and vice versa. Based on this, the enzyme activity can be calculated. The substrate for SOD activity detection is a mixture of 150μL of 0.05mol / L phosphate buffer solution, 20μL of distilled water, 30μL of 130mmol / LMet solution, 30μL of 750μmol / LNBT solution, 30μL of 100μmol / LEDTA-Na2 solution and 30μL of 20μmol / L riboflavin solution. After mixing, use it immediately, and then drop 10μL of SOD crude enzyme solution (set up 2 control tubes, and the control tubes use buffer instead of enzyme solution). Place one of the control tubes in the dark, and the rest are placed under 4000Lx fluorescent light for 20min. The SOD activity determination of the same replicate material uses the same batch of substrates to avoid experimental errors caused by substrate concentration errors. The OD value was measured at 560 nm and 25 °C using an enzyme reader (Synergy H1, USA).

[0041] Unit SOD activity (U) definition: 50% inhibition of NBT photoreduction is expressed as one unit of enzyme activity.

[0042] ; Among them, the total SOD activity was expressed as enzyme units per gram of fresh mass of sample (U / g); Ack: absorbance of the control tube under illumination; AE: absorbance of the sample tube; V: total volume of sample solution (mL); Vt: sample volume used in the determination (mL); W: fresh weight of sample (g).

[0043] The results of SOD activity detection of wheat grains of various varieties are shown in Table 1: The PCR amplification products obtained from 125 winter wheat varieties are divided into two types according to their sizes: one is 643 bp; the other is 1106 bp. The electrophoresis diagram of some samples is shown in Figure 1(Among them, lanes 1-10 samples correspond to the wheat varieties in Table 1; lane 1: Shiyou 17; lane 2: Huaimai 18; lane 3: Xiaoyan 22; lane 4: Zimai 12; lane 5: Zhongyu 5; lane 6: Shan 715; lane 7: Jimai 21; lane 8: Linkang 12; lane 9: Zhoumai 12; lane 10: Neixiang 188). Among the 125 wheat varieties: 67 varieties contain the allele TaSOD-B2a, and the average SOD activity is 1746.27U / g; 58 varieties contain the allele TaSOD-B2b, and the average SOD activity is 1828.67U / g; the SOD activity of wheat varieties containing the allele TaSOD-B2b is higher than that of wheat varieties containing the allele TaSOD-B2a, and there is a significant difference between the two (P<0.05).

[0044] The results showed that among the 67 wheat varieties containing the TaSOD-B2a allele, 27 were varieties with low SOD activity, that is, the accuracy rate of identifying wheat with low SOD activity by using the allele TaSOD-B2a was 40.3%; among the 58 wheat varieties containing the TaSOD-B2b allele, 33 were varieties with high SOD activity, that is, the accuracy rate of identifying wheat with high SOD activity by using the allele TaSOD-B2b was 56.9%.

[0045] Table 1. PCR test results and SOD activity test results of various wheat varieties

[0046]

[0047]

[0048] Note: “√” indicates the presence of this band in the PCR product.

[0049] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. A primer composition, characterized in that: The primer combination consists of primer pair 1 and primer pair 2; the primer pair 1 consists of the single-stranded DNA shown in SEQ ID No.1 and the single-stranded DNA shown in SEQ ID No.2; the primer pair 2 consists of the single-stranded DNA shown in SEQ ID No.3 and the single-stranded DNA shown in SEQ ID No.

4.

2. A reagent and / or kit for identifying superoxide dismutase activity in plant seeds, characterized in that: The reagent and / or kit contains the primer composition according to claim 1.

3. The reagent and / or kit according to claim 2, characterized in that: The final concentration of each primer in the primer pair 1 and / or the primer pair 2 in the PCR reaction system is 10 -7 mol / L.

4. Any of the following uses of the primer composition according to claim 1 and / or the reagent and / or kit according to claim 2 or 3: P1. Application in identification or auxiliary identification of SOD activity in wheat to be tested; P2. Application in detecting or assisting in detecting wheat SOD activity; P3. Application in the preparation and identification of products or in assisting the identification of wheat SOD activity; P4. Application in the preparation of products for detecting or assisting in detecting the activity of wheat SOD to be tested; P5. Application in the preparation of products for detecting or assisting in detecting the TaSOD-B2 genotype of wheat to be detected; P6. Application in breeding or improvement of wheat SOD activity phenotype.

5. Use of the primer composition according to claim 1 or the reagent and / or kit according to claim 2 or 3 in detecting or assisting in detecting whether the wheat to be tested is high SOD activity wheat or low SOD activity wheat; The SOD activity in the low SOD activity wheat is less than or equal to 1746.27 U / g; The SOD activity in the high SOD activity wheat is greater than or equal to 1828.67 U / g.

6. A method for detecting or assisting in detecting the SOD activity of wheat to be tested, comprising the following steps: The genomic DNA of the wheat to be tested is subjected to PCR amplification using the primer composition described in claim 1 and / or the reagent and / or kit described in claim 2 or 3 to obtain a PCR amplification product; and the SOD activity of the wheat to be tested is detected based on the size of the PCR amplification product.

7. A method for detecting or assisting in detecting that wheat to be tested contains TaSOD-B2a haplotype and / or TaSOD-B2b haplotype, comprising the following steps: Using the primer pair 1 and the primer pair 2 described in claim 1, respectively, to perform PCR amplification on the genomic DNA of the wheat to be tested to obtain PCR amplification products; according to the length of the DNA fragment of the PCR amplification product band, confirm whether the wheat to be tested contains TaSOD-B2a haplotype and / or TaSOD-B2b haplotype; The wheat to be tested whose PCR amplification product contains a DNA fragment of 643 bp in length contains the TaSOD-B2a haplotype, and the wheat to be tested whose PCR amplification product contains a DNA fragment of 1106 bp in length contains the TaSOD-B2b haplotype; The TaSOD-B2a haplotype corresponds to nucleotides 2689-3331 of SEQ ID No. 5 in the sequence listing, and the TaSOD-B2b haplotype corresponds to nucleotides 2689-3794 of SEQ ID No. 6 in the sequence listing.

8. A method for cultivating wheat with low SOD activity, comprising the following steps: breeding with plant materials of group A as parents to cultivate wheat with low SOD activity; The plant material of group A meets the following conditions: the amplification product obtained by PCR amplification of genomic DNA using the primer pair 1 in claim 1 contains a DNA fragment of 643 bp in size, and the amplification product obtained by PCR amplification of genomic DNA using the primer pair 2 in claim 1 does not contain a DNA fragment of 1106 bp in size; one chain of the DNA fragment of 643 bp corresponds to nucleotides 2689-3331 of SEQ ID No.5 in the sequence listing; one chain of the DNA fragment of 1106 bp corresponds to nucleotides 2689-3794 of SEQ ID No.6 in the sequence listing; the SOD activity in the wheat grains with low SOD activity is less than or equal to 1746.27 U / g.

9. A method for cultivating wheat with high SOD activity, comprising the following steps: using group B wheat as a parent to perform wheat breeding to cultivate wheat with high SOD activity; The group B wheat meets the following conditions: the amplification product obtained by PCR amplification of genomic DNA using the primer pair 2 in claim 1 contains a fragment of 1106 bp in size, and the amplification product obtained by PCR amplification of genomic DNA using the primer pair 1 in claim 1 does not contain a DNA fragment of 643 bp in size; one chain of the DNA fragment of 643 bp corresponds to nucleotides 2689-3331 of SEQ ID No.5 in the sequence listing; one chain of the DNA fragment of 1106 bp corresponds to nucleotides 2689-3794 of SEQ ID No.6 in the sequence listing; the SOD activity in the grains of the high SOD activity wheat is greater than or equal to 1828.67 U / g.

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