White-grain wheat pre-harvest germination resistance gene TaMyb10-94E and application thereof

Abstract

The invention discloses a white wheat pre-harvest germination resistance gene TaMyb10-94E and an application thereof. The TaMyb10-94E gene is cloned from the 3D chromosome of the baryblue wheat, and a stable transgenic technology proves that overexpression of the TaMyb10-94E does not cause red color of wheat grains like overexpression of other TaMyb10 genes, and meanwhile, the TaMyb10-94E gene has the functions of enhancing the dormancy of the grains and reducing the germination rate of the grains. The result shows that the special variation of the TaMyb10-94E gene derived from the optionally selected lump wheat can endow the wheat with sprouting resistance under the condition that red grains are not generated. The method has an important application value for cultivating white-grain pre-harvest sprouting resistant wheat.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a white-grain wheat spike sprouting resistance gene TaMyb10-94E and an application thereof. Background Art

[0002] Wheat is an important food crop in the world. About 35% to 40% of the world's population relies on wheat as their staple food. Ear sprouting refers to the phenomenon that wheat seeds germinate on the ear when they encounter continuous wet weather before harvest. Ear sprouting will cause the activity of related hydrolases in wheat grains to increase rapidly, degrading the storage materials in the grains, seriously affecting the quality of wheat. It will also affect the storage of wheat and the germination rate of sowing in the next year, causing great economic losses to wheat production. In long-term production practice, people have found that there is a close relationship between wheat seed coat color and ear sprouting. Common wheat seed coat colors are mainly divided into red and white, and the former is generally more resistant to ear sprouting than the latter. In the wheat areas of Sichuan, Anhui, and Jiangsu, where there is abundant rainfall and serious ear sprouting, red-skinned wheat is often planted. However, white-skinned wheat is more popular in production because it has the characteristics of thin skin, high relative endosperm content, and high commercial wheat flour yield. Therefore, the discovery and utilization of white-grain wheat ear sprout resistance genes are of great value. Summary of the invention

[0003] The purpose of the present invention is to provide a white-grain wheat spike sprouting resistance gene TaMyb10-94E and an application thereof.

[0004] The wheat TaMyb10 gene activates the synthesis pathway of flavonoids and ABA hormones, which can simultaneously regulate the color and germination rate of wheat grains. Most of the wheat grains with this gene on the wheat chromosome 3D are red. The white-grain wheat Suining Tuotuo wheat is a farmer's variety with a germination index of 10.1%, which is a material resistant to ear germination. In the F2 generation population of the hybrid of Suining Tuotuo wheat (white grain) × China Spring (red grain), individuals with extreme red and white grains were selected for mixed sequencing to observe the co-segregation of grain color and TaMyb10 gene allele variation. It was found that the mixed samples of white-grain wheat materials were all TaMyb10-94E gene types, and the mixed samples of red-grain wheat materials were heterozygous, indicating that the TaMyb10-94E gene does not cause wheat grains to turn red. Therefore, the special variation of the TaMyb10-94E gene from the 3D chromosome of Suining Tuotuo wheat can confer wheat ear germination resistance without producing red grains.

[0005] In order to achieve the purpose of the present invention, in the first aspect, the present invention provides a white-grain wheat spike sprout resistance gene TaMyb10-94E (from Tuotuo wheat in Suining, Sichuan), which is a gene encoding the following protein (a) or (b):

[0006] (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 1;

[0007] (b) A protein derived from (a) with equivalent functions, wherein one or more amino acids are substituted, deleted or added to the sequence shown in SEQ ID NO: 1.

[0008] The nucleotide sequence of gene TaMyb10-94E is:

[0009] i) the nucleotide sequence shown in SEQ ID NO: 2;

[0010] ii) a nucleotide sequence in which one or more nucleotides are substituted, deleted and / or added to the nucleotide sequence shown in SEQ ID NO: 2 and the nucleotide sequence expresses a protein with the same function;

[0011] iii) a nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO: 2 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization in a 0.1×SSPE solution containing 0.1% SDS or a 0.1×SSC solution containing 0.1% SDS at 65°C, and the membrane is washed with the solution; or

[0012] iv) a nucleotide sequence that has more than 90% homology with the nucleotide sequence of i), ii) or iii) and expresses a protein with the same function.

[0013] In a second aspect, the present invention provides biological materials containing the gene TaMyb10-94E, wherein the biological materials include but are not limited to recombinant DNA, expression cassettes, transposons, plasmid vectors, viral vectors, engineered bacteria or transgenic cell lines.

[0014] In a third aspect, the present invention provides any of the following uses of the gene TaMyb10-94E or a biological material containing the gene:

[0015] (1) Used to regulate wheat grain color;

[0016] (2) Used to regulate wheat grain germination rate;

[0017] (3) Used in plant breeding or plant germplasm improvement;

[0018] (4) Used for preparing transgenic plants.

[0019] Furthermore, the application includes:

[0020] 1) making wheat contain gene TaMyb10-94E; or

[0021] 2) causing wheat to express the protein encoded by the gene TaMyb10-94E; or

[0022] 3) Overexpressing the gene TaMyb10-94E in wheat;

[0023] Thus, white-grain wheat with resistance to ear sprouting is obtained.

[0024] In a fourth aspect, the present invention provides a method for constructing white-grain wheat resistant to ear sprouting, the method comprising: overexpressing the gene TaMyb10-94E in wheat by genetic engineering means.

[0025] Furthermore, the overexpression method may be selected from the following 1) to 5), or an optional combination:

[0026] 1) by introducing a plasmid carrying the gene;

[0027] 2) by increasing the copy number of the gene on the plant chromosome;

[0028] 3) by changing the promoter sequence of the gene on the plant chromosome;

[0029] 4) by operably linking a strong promoter to the gene;

[0030] 5) by introducing enhancers;

[0031] 6) The TaMyb10 gene in wheat was transformed into the TaMyb10-94E gene through gene editing.

[0032] The expression vector carrying the target gene can be introduced into plant cells by conventional biotechnology methods such as Ti plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, etc. (Weissbach, 1998, Method for Plant Molecular Biology VIII, Academy Press, New York, pp. 411-463; Geiserson and Corey, 1998, Plant Molecular Biology, pp. 2 nd Edition).

[0033] Preferably, the expression vector containing the gene is transformed into Agrobacterium, and the gene is transferred into the plant by Agrobacterium-mediated transformation, so that the gene is overexpressed in the plant.

[0034] In a fifth aspect, the present invention provides the use of the transgenic wheat obtained according to the method in plant breeding.

[0035] Further, breeding methods include but are not limited to transgenic, hybridization, backcrossing, selfing or asexual reproduction.

[0036] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0037] The present invention identifies the TaMyb10-94E gene from the 3D chromosome from Suining Tuotuo wheat for the first time. Compared with other wheat TaMYB10 proteins including Chinese Spring, the 94th amino acid of the TaMYB10-94E protein in Tuotuo wheat changes from glycine (G) to glutamic acid (E). Experiments have shown that the present invention found that the TaMyb10-94E gene and the protein it encodes can control wheat grain color and spike sprout resistance, and further verified its function, which can be used to construct white-grained spike sprout-resistant wheat. This gene can be used in plant genetic engineering, gene editing technology and molecular breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The figure is a comparison of the nucleic acid sequences of the TaMyb10-94E of Suining Tuotuo wheat and other wheat TaMyb10 genes in a preferred embodiment of the present invention.

[0039] Figure 2 The figure shows the alignment of TaMyb10-94E of Tuotuo wheat in a preferred embodiment of the present invention with TaMyb10 protein sequences of other wheats represented by wheat Chinese Spring (protein-mc is Chinese Spring, protein-mt is Suining Tuotuo wheat).

[0040] Figure 3 This is the expression of the TaMyb10 gene in Tuotuo wheat seeds in a preferred embodiment of the present invention.

[0041] Figure 4 This is the co-segregation identification of the grain color and TaMyb10 gene allelic variation of the hybrid offspring of Suining Tuotuo wheat (white grain) × China Spring (red grain) in the preferred embodiment of the present invention.

[0042] Figure 5 This is the identification result of transgenic wheat overexpressing TaMyb10-94E in a preferred embodiment of the present invention.

[0043] Figure 6 This is the germination rate of TaMyb10-94E-OE transgenic wheat grains in a preferred embodiment of the present invention.

[0044] Figure 7 The grain color comparison of TaMyb10-94E-OE transgenic wheat (2# and 12#) and TaMyb10-OE transgenic wheat (58-4#) in the preferred embodiment of the present invention is shown. DETAILED DESCRIPTION

[0045] The present invention provides a white-grain wheat (Suining Tuotuo wheat) spike sprout resistance gene TaMyb10-94E and a cloning method thereof, including extraction, cloning, and functional analysis of the TaMyb10-94E gene. The white-grain wheat spike sprout resistance gene TaMyb10-94E obtained from Suining Tuotuo wheat has a specific base sequence, and its protein is named TaMYB10-94E protein. Compared with the known wheat TaMYB10 protein, it has a different amino acid sequence and has characteristics that other TaMYB10 proteins do not have (does not produce red grain traits), has different types of functions, and is a special type that has not been found in the relevant field. Specifically, it is as follows (a) or (b):

[0046] (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 1;

[0047] (b) A protein derived from SEQ ID NO: 1 which is related to plant seed color and ear germination and in which one or more amino acid residues of the amino acid sequence of SEQ ID NO: 1 are substituted and / or deleted and / or added.

[0048] In order to facilitate the purification and detection of the protein in (a), a tag as shown in Table 1 can be connected to the amino terminus or carboxyl terminus of the protein shown in (a).

[0049] Table 1 Tag sequences

[0050] Label Residue sequence Poly-Arg 5-6 (usually 5) RRRRR Poly-His 2-10 (usually 6) HHHHHH FLAG 8 DYKDDDDK Strep-tag II 8 WSHQ c-myc 10 EQKLISEEDL

[0051] The protein in (b) above can be synthesized artificially, or its coding gene can be synthesized first and then expressed biologically. The coding gene of the protein in (b) above can be obtained by deleting one or several codons of amino acid residues in the coding gene of TaMYB10-94E protein, and / or performing missense mutation of one or several base pairs, and / or connecting the coding sequence of the tag shown in Table 1 to its 5′ end and / or 3′ end.

[0052] The gene encoding the TaMYB10-94E protein (TaMyb10-94E gene) also falls within the protection scope of the present invention.

[0053] The TaMyb10-94E gene is a DNA molecule described in any one of the following (1)-(3):

[0054] (1) a DNA molecule whose coding region is shown in SEQ ID NO: 2;

[0055] (2) a DNA molecule that hybridizes to the DNA sequence defined in (1) under stringent conditions and encodes a protein associated with plant seed color;

[0056] (3) A DNA molecule having 90% or more homology with the DNA sequence defined in (1) or (2) and encoding a protein associated with plant height.

[0057] The stringent conditions mentioned above may be using a solution of 0.1×SSPE (or 0.1×SSC), 0.1% SDS, hybridizing at 65° C. and washing the membrane in a DNA or RNA hybridization experiment.

[0058] Those skilled in the art can mutate the nucleotide sequence encoding the TaMYB10-94E protein of the present invention by known methods, such as directed evolution and point mutation. Those artificially modified nucleotides having 75% or higher identity with the nucleotide sequence of the TaMYB10-94E protein isolated from the present invention are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention as long as they encode the TaMYB10-94E protein.

[0059] The present invention also provides a recombinant expression vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the TaMyb10-94E gene.

[0060] The present invention also protects the use of any of the above-mentioned TaMYB10-94E proteins, or any of the above-mentioned nucleic acid molecules, or an expression box, a recombinant vector or a recombinant microorganism containing any of the above-mentioned nucleic acid molecules in regulating wheat grain color and ear sprout resistance, as well as in cultivating white-grain transgenic wheat with resistance to ear sprout.

[0061] Existing plant expression vectors can be used to construct a recombinant expression vector containing the TaMyb10-94E gene. The plant expression vector includes a binary Agrobacterium vector and a vector that can be used for plant microprojectile bombardment. When the TaMyb10-94E gene is used to construct a recombinant expression vector, any enhanced, constitutive, tissue-specific or inducible promoter can be added before its transcription start nucleotide, which can be used alone or in combination with other plant promoters; in addition, when the TaMyb10-94E gene is used to construct a recombinant expression vector, an enhancer can also be used, including a translation enhancer or a transcription enhancer. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The sources of the translation control signal and the start codon are extensive and can be natural or synthetic. The translation start region can come from the transcription start region or the structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that express enzymes or luminescent compounds that can produce color changes in plants, antibiotic resistance markers, or chemical resistance marker genes.

[0062] The recombinant expression vector can be specifically a recombinant expression vector obtained by replacing the fragment between the BamHI and PmlI restriction sites of the pUBI:cas vector with the DNA molecule shown in SEQ ID NO:2.

[0063] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the examples are all based on conventional experimental conditions, such as Sambrook et al. Molecular Cloning Laboratory Manual (Sambrook J & Russell DW, Molecular Cloning: a Laboratory Manual, 2001), or the conditions recommended by the manufacturer's instructions.

[0064] The transcriptome sequencing in the following examples was performed three times, and the results were averaged.

[0065] For the local wheat variety Sichuan Suining Tuotuo wheat, please refer to Zhou Yong. Evaluation of ear sprout resistance and genome-wide association analysis of Chinese wheat local varieties[D]. Sichuan Agricultural University, 2017; the public can obtain this biological material from the Wheat Research Institute of Sichuan Agricultural University.

[0066] Wheat other TaMyb10 gene (sequence as shown in SEQ ID NO: 3) overexpressing transgenic line 58-4# can be found in Lang J, Fu Y, Zhou Y, Cheng M, Deng M, Li M, Zhu T, Yang J, Guo X, Gui L, Li L, Chen Z, Yi Y, Zhang L, Hao M, Huang L, Tan C, Chen G, Jiang Q, Qi P, Pu Z, Ma J, Liu Z, Liu Y, Luo MC, Wei Y, Zheng Y, Wu Y, Liu D, Wang J. Myb10-D confers PHS-3D resistance to pre-harvest sprouting by regulating NCED in ABA biosynthesis pathway of wheat[J]. New phytologist, 2021, 230(5): 1940-52. The public can obtain the biological material from the Wheat Research Institute of Sichuan Agricultural University.

[0067] The pUBI-CAMBIA3301 vector can be found in Zhang L, He G, Li Y, et al. PIL transcription factors directly interact with SPLs and repress tillering / branching in plants[J]. New Phytologist, 2022, 233(3): 1414-1425. The public can obtain this biological material from the Wheat Research Institute of Sichuan Agricultural University.

[0068] Example 1 Obtaining TaMYB10-94E protein and its encoding gene

[0069] Total RNA was extracted from the common wheat local variety Suining Tuotuomai and reverse transcribed into cDNA. PCR amplification was performed using a primer pair consisting of TaMYB10-F: 5'-ATGGGGAGGAAGCCATGCTGCGCCA-3' and TaMYB10-R: 5'-CTAGCAAAGCCACGCCAACTCCAGG-3'. The reaction procedure was: 94℃2min; 94℃30sec, 60℃30sec, 68℃1min, 34 cycles; 68℃10min. The PCR amplification product of about 798bp was recovered and sent to Qingke Biotechnology Co., Ltd. for sequencing. The sequencing results showed that the DNA sequence shown in SEQ ID NO:2 was obtained from the cDNA amplification product, and the protein encoded by it was shown in SEQID NO:1.

[0070] The protein shown in SEQ ID NO:1 was named TaMYB10-94E protein. The gene encoding TaMYB10-94E protein was named TaMyb10-94E gene, and its open reading frame was shown in SEQ ID NO:2.

[0071] Compared with other wheat TaMYB10 genes including Chinese Spring, the 281st base of the TaMyb10-94E gene coding region in Tuotuo wheat changes from G to A ( Figure 1 ).

[0072] Compared with other wheat TaMYB10 proteins including Chinese Spring, the 94th amino acid of TaMYB10-94E protein in Tuotuo wheat changes from glycine (G) to glutamic acid (E) ( Figure 2 ).

[0073] Example 2 TaMyb10-94E gene co-segregates with grain color

[0074] 1. The red-grain wheat Chinese Spring was hybridized with the white-grain wheat Suining Tuotuo wheat. The resulting seeds were recorded as F0 and were propagated to the F3 generation through field self-pollination.

[0075] 2. Take leaves from each wheat plant of the F3 generation, extract total DNA using the CTAB method, and use it as a template. Use the primer pair composed of TaMYB10-F and TaMYB10-R mentioned above for PCR amplification. Recover the PCR amplification product of about 798 bp and send it to Qingke Biotechnology Co., Ltd. for sequencing.

[0076] 3. Harvest the F3 generation, and soak each wheat grain with vanillin reagent. Vanillin will undergo a condensation reaction with resorcinol or phloroglucinol, which are monomers of proanthocyanidins and catechins in the grains. The product will form red positive carbon ions under the action of concentrated acid, which will make the grains appear in different colors. Corresponding to the sequencing results of step 2, the results showed that the Chinese spring and F3 red-grain wheat are both TaMYB10 genotypes, and Suining Tuotuo wheat (TTM) and F3 white-grain wheat are both TaMyb10-94E genotypes ( Figure 3 ), indicating that the TaMyb10-94E gene was co-segregated with white grains.

[0077] Example 3 Analysis of TaMyb10-94E gene expression pattern

[0078] 1. Cultivate Suining Tuotuo wheat in an artificial climate chamber for 3 months, and take samples at different days (5d, 7d, 10d, 12d, 15d, and 20d) after flowering.

[0079] 2. Extract the total RNA from the samples obtained at each time point in step 1 and send them to Beijing Berry Genomics Co., Ltd. (berrygenomics.com) for next-generation transcriptome sequencing. The total data volume of a single sample is 6G. Kallisto was used to calculate the TPM of the gene and obtain the expression trend of TaMyb10-94E in Tuotuo wheat. The results are as follows Figure 4 The results showed that the expression level of TaMyb10-94E reached peak values ​​on the 5th and 10th days, and dropped to 0 on the 20th day.

[0080] Example 4 Application of TaMyb10-94E gene in regulating wheat grain color and ear sprout resistance

[0081] 1. Construction of recombinant expression vector

[0082] The fragment between the BamHI and PmlI restriction sites of the pUBI-CAMBIA3301 vector was replaced with the DNA molecule shown in SEQ ID NO: 2 to obtain a recombinant expression vector (verified by sequencing).

[0083] 2. Obtaining recombinant Agrobacterium

[0084] The recombinant expression vector obtained in step 1 was introduced into Agrobacterium tumefaciens EHA105 by heat shock transformation method to obtain recombinant Agrobacterium.

[0085] 3. Acquisition and identification of transgenic wheat

[0086] The recombinant Agrobacterium obtained in step 2 was used to transform the immature embryos of the wheat variety Fielder (white grain, non-anti-ear sprouting variety) using the Agrobacterium-mediated genetic transformation method to obtain the transgenic wheat plant TaMyb10-94E-OE. The Fielder genome does not contain the TaMyb10-94E gene, so the leaf DNA of the control Fielder (WT) and the transgenic wheat was extracted and PCR amplified using the primer pair composed of TaMYB10-F and TaMYB10-R. The results are as follows: Figure 5 As shown, the TaMyb10-94E gene can be amplified in the transgenic strain, but no band is amplified in the WT.

[0087] 4. Phenotypic analysis of transgenic plants

[0088] Plants to be tested: WT, TaMyb10-94E-OE transgenic lines obtained in step 3 (2# and 12#), and transgenic line 58-4# obtained by overexpressing other TaMyb10 genes of wheat in Fielder background.

[0089] Wheat ears were harvested at the waxy stage, naturally air-dried indoors, threshed after 7 days, and their moisture content was measured to 10%-12%. After disinfection and rinsing with 0.5% NaClO, they were placed in a culture dish (with moist filter paper at the bottom of the culture dish). The number of germinated seeds was counted from the second day of the experiment, and the germinated seeds were removed and counted continuously for 7 days. The germination rate was calculated as follows: germination rate = (number of germinated seeds / total number of seeds) × 100%. The results are as follows: Figure 6 As shown, the 7-d grain germination rate of WT was over 80%, while overexpression of TaMyb10-94E significantly inhibited the grain germination rate.

[0090] After harvesting the mature seeds, they were treated with vanillin and NaOH solution. NaOH reacts with flavonols and dihydroflavonoids to turn brown or yellow. Figure 7 Overexpression of TaMyb10-94E (2# and 12#) caused white wheat grains, and overexpression of other TaMyb10 genes (58-4#) caused red wheat grains.

[0091] In summary, there are common wheats that do not contain the TaMyb10 gene in natural wheat materials, such as Fielder (white grains, no resistance to ear sprouting). Studies have found that transgenic wheat overexpressing the TaMyb10-94E gene in the Fielder background has white grains and is resistant to ear sprouting ( Figure 6 and Figure 7 ). However, transgenic wheat overexpressing other TaMyb10 genes in the Fielder background had red grains and ear sprout resistance, indicating that the TaMyb10-94E gene has the function of producing ear sprout resistance instead of red grains. The gene TaMyb10-94E regulates wheat grain color and grain germination.

[0092] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. White-grain wheat spike sprout resistance gene TaMyb10-94E, characterized in that: It is a gene encoding the following protein (a) or (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 1; (b) A protein derived from (a) with equivalent functions, wherein one or more amino acids are substituted, deleted or added to the sequence shown in SEQ ID NO:

1.

2. The gene TaMyb10-94E according to claim 1, characterized in that Its nucleotide sequence is: i) the nucleotide sequence shown in SEQ ID NO: 2; ii) a nucleotide sequence in which one or more nucleotides are substituted, deleted and / or added to the nucleotide sequence shown in SEQ ID NO: 2 and the nucleotide sequence expresses a protein with the same function; iii) a nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO: 2 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization in a 0.1× SSPE solution containing 0.1% SDS or a 0.1× SSC solution containing 0.1% SDS at 65°C, and the membrane is washed with the solution; or iv) a nucleotide sequence that has more than 90% homology with the nucleotide sequence of i), ii) or iii) and expresses a protein with the same function.

3. A biological material containing the gene TaMyb10-94E according to claim 1 or 2, characterized in that: The biological material is recombinant DNA, expression cassette, transposon, plasmid vector, virus vector or engineered bacteria.

4. Any of the following uses of the gene TaMyb10-94E according to claim 1 or 2 or the biomaterial according to claim 3: (1) Used to regulate wheat grain color; (2) Used to regulate wheat grain germination rate; (3) Used in plant breeding or plant germplasm improvement; (4) Used for preparing transgenic plants.

5. The use according to claim 4, characterized in that: The applications include: 1) making wheat contain gene TaMyb10-94E; or 2) causing wheat to express the protein encoded by the gene TaMyb10-94E; or 3) Overexpressing the gene TaMyb10-94E in wheat; Thus, white-grain wheat with resistance to ear sprouting is obtained.

6. A method for constructing white-grain wheat with resistance to ear sprouting, characterized in that: The method comprises: using genetic engineering means to overexpress the gene TaMyb10-94E of claim 1 or 2 in wheat; The overexpression method is selected from the following 1) to 5), or an optional combination: 1) by introducing a plasmid carrying the gene; 2) by increasing the copy number of the gene on the plant chromosome; 3) by changing the promoter sequence of the gene on the plant chromosome; 4) by operably linking a strong promoter to the gene; 5) by introducing enhancers; 6) The TaMyb10 gene in wheat was transformed into the TaMyb10-94E gene through gene editing.

7. The method according to claim 6, characterized in that The expression vector containing the gene is transformed into Agrobacterium, and the gene is transferred into plants through the Agrobacterium-mediated transformation method, so that the gene is overexpressed in the plant.

8. Use of the transgenic wheat obtained according to the method of claim 6 or 7 in plant breeding.

9. The use according to claim 8, characterized in that: Breeding methods include transgenic, hybridization, backcrossing, selfing or asexual reproduction.

Images