Soybean GmSMS6 protein and application of coding gene thereof in regulation and control of soybean grain quality traits

Abstract

The invention provides application of soybean GmSMS6 protein and a coding gene thereof in regulation and control of soybean grain quality traits. The genome sequence of the GmSMS6 is as shown in SEQ ID No. 1, and the coded amino acid sequence is as shown in SEQ ID No. 2. Gene editing is carried out on soybean Zp661 through a CRSIPR-Cas9 system, homozygous mutant plants Z1 and Z2 with the seed protein content obviously increased and the oil content obviously reduced compared with wild soybean Zp661 are obtained, and it is proved that soybean GmSMS6 has the function of regulating and controlling the quality of soybean seeds. The invention provides important gene resources and theoretical guidance for soybean molecular breeding and germplasm innovation.

Description

Technical Field

[0001] This invention belongs to the field of mutation or genetic engineering, specifically relating to the application of soybean GmSMS6 protein and its encoding gene in regulating soybean grain quality traits. Background Technology

[0002] Soybeans Soybean Soybean is an important dual-purpose crop, providing humans and animals with abundant oils, proteins, and other nutrients. Although my country is the origin of soybeans, soybean breeding has developed slowly in recent years, and the yield per unit area lags behind that of major soybean-producing countries in the world. Currently, China's soybean self-sufficiency rate is severely insufficient, with a high dependence on imports and a self-sufficiency rate of less than 20%.

[0003] Soybean seed protein and oil content are quantitative traits controlled by multiple genes and are also influenced by environmental factors. Multiple factors and regulatory pathways jointly determine the protein and oil content of soybean seeds. Although some studies have reported gene loci regulating soybean protein and oil content, the functional identification of key genes and the regulatory network remain unclear. Elucidating the molecular mechanisms of soybean seed protein and oil content formation and revealing the synergistic regulatory network between them is an important theoretical foundation for creating high-yielding and high-quality soybean varieties. In recent years, with the continuous advancement of soybean genomics and modern molecular biology research, techniques such as gene-directed modification have enabled the precise regulation of soybean seed protein and oil content to achieve the breeding goals of high-yielding and high-quality soybean varieties. Summary of the Invention

[0004] The technical problem to be solved by this invention is how to use biological materials to regulate the quality of plants, especially soybean seeds.

[0005] Therefore, the present invention provides a method for regulating plant seed traits, characterized in that the method includes regulating the seed traits of a target plant by gene knockout, wherein the gene knockout is the knockout of the gene encoding a target protein in the target plant, and the target plant contains the gene encoding the target protein. The target protein is GmSMS6, and is any one of the following: A1) The amino acid sequence is that of the protein shown in sequence 2. A2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein described in A1), which has more than 80% identity with the protein shown in A1) and can regulate plant grain traits. A3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of A1) or A2).

[0006] The present invention also provides a method for cultivating plants with altered seed traits, comprising obtaining a target plant with altered seed traits by gene knockout, wherein the target plant contains the gene encoding the aforementioned protein.

[0007] The protein is derived from soybeans.

[0008] In the above method, the genome of the target plant contains a DNA molecule whose nucleotide sequence is sequence 1 in the sequence listing, and the gene knockout includes performing any of the following operations on the genome of the target plant: B1) Delete nucleotides from position 342 to 344 of sequence 1 in the sequence listing of the target plant, and insert an adenine ribonucleotide between positions 491 and 492.

[0009] B2) Delete nucleotides 336-340 from sequence 1 in the sequence listing of the target plant.

[0010] Gene knockout refers to the phenomenon of inactivating a specific target gene through homologous recombination. Gene knockout inactivates a specific target gene by altering its DNA sequence.

[0011] The target sequence for gene knockout may be sgRNA1:5'-GTCGCCTACAAGAACGTAAT-3' and / or sgRNA2:5'-CAAGCTCCTCGACACGCGCC-3'.

[0012] The gene knockout includes introducing a gene knockout vector into the target plant with the target sequences 5'-GTCGCCTACAAGAACGTAAT-3' and / or sgRNA2:5'-CAAGCTCCTCGACACGCGCC-3'.

[0013] This invention also protects the application of biological materials, said application being any of the following: C1) The application of the biomaterials described herein in regulating plant grain traits and / or preparing products that regulate the grain traits of target plants. C2) The application of the biomaterials described herein in plant breeding and / or the preparation of plant breeding products; The biological material is a protein, a substance that regulates the expression of the gene encoding the protein, or a substance that regulates the content of the protein. The protein is GmSMS6, and is any one of the following: D1) The amino acid sequence is the protein shown in sequence 2. D2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein described in D1), which has more than 80% identity with the protein shown in D1) and can regulate plant grain traits. D3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of D1) or D2).

[0014] In the aforementioned proteins, the protein tag refers to a polypeptide or protein fused with the target protein using in vitro DNA recombination technology for expression, detection, tracing, and / or purification of the target protein. The protein tag may be a Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, and / or SUMO tag, etc.

[0015] In the above-mentioned proteins, identity refers to the identity of the amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, using blastp as the program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as the matrix, setting the Gap existence cost, Per residue gap cost, and Lambda ratio to 11, 1, and 0.85 (default values) respectively, and performing an identity search on a pair of amino acid sequences to calculate the identity value (%), then the identity value can be obtained.

[0016] In the aforementioned proteins, the 80% or more identity can be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99%, or 100% identity.

[0017] The amino acid sequence of the above protein is sequence 2 in the sequence listing, consisting of 262 amino acid residues, and it is named GmSMS6 protein or protein GmSMS6. Its encoding gene is... GmSMS6 Gene. Sequence 2 (SEQ ID No. 2) is as follows: MAASAPTPREEFVYMAKLAEQAERYEEMVEFMEKVSASAESEELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNEDHVAVIRDYRSKIEAELSNICDGILKLLDTRLVPSAASGDSKVFYLKM KGDYHRYLAEFKTGADRKEAAESTLSAYKAAQDIANTELPPTHPIRLGLALNFSVFYYEILNSPDRACSLAKQAFDEAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMQDDGADEIKEAAPKGDGEQN.

[0018] In the above text, the substance referred to is any one of the following: E1) Nucleic acid molecules that inhibit, reduce, or downregulate the expression of the genes encoding the above proteins. E2) expresses the gene encoding the nucleic acid molecule described in E1). E3) contains the expression cassette of the gene described in E2). E4) A recombinant vector containing the gene described in E2), or a recombinant vector containing the expression cassette described in E3). E5) Recombinant microorganisms containing the gene described in E2), or recombinant microorganisms containing the expression cassette described in E3), or recombinant microorganisms containing the recombinant vector described in E4). E6) A transgenic plant cell line containing the gene described in E2), or a transgenic plant cell line containing the expression cassette described in E3), or a transgenic plant cell line containing the recombinant vector described in E4). E7) Transgenic plant tissue containing the gene described in E2), or transgenic plant tissue containing the expression cassette described in E3), or transgenic plant tissue containing the recombinant vector described in E4). E8) A transgenic plant organ containing the gene described in E2), or a transgenic plant organ containing the expression cassette described in E3), or a transgenic plant organ containing the recombinant vector described in E4).

[0019] Furthermore, in the biological material, the recombinant microorganism (E5) can specifically be yeast, bacteria, algae, and fungi.

[0020] Furthermore, the recombinant microorganism may be Agrobacterium. Specifically, Agrobacterium is EHA105.

[0021] Furthermore, in the biological material, the plant tissue described in E7) may be derived from roots, stems, leaves, flowers, fruits, seeds, pollen, embryos and / or anthers.

[0022] Furthermore, in the biological material, the transgenic plant organs described in E8) can be the roots, stems, leaves, flowers, fruits, and seeds of the transgenic plant.

[0023] In the above text, the regulation of plant grain traits refers to increasing the protein content of plant grains and / or decreasing the oil content of plant grains.

[0024] The plant mentioned above is any one of the following: F1) dicotyledonous plants, F2) Rosales plants, F3) legumes, F4) soybeans, F5) soybean.

[0025] This invention utilizes CRISPR-Cas9 technology to target the soybean recipient plant Zp661. GmSMS6 Gene editing yielded... GmSMS6 Homozygous mutants of the gene Z1 and Z2. Compared with wild-type soybean Zp661, GmSMS6 The soybean seeds of the gene-mutated plants Z1 and Z2 showed significantly increased protein content and significantly decreased oil content. This indicates... GmSMS6 Genes negatively regulate soybean seed protein content and also affect seed oil content. This invention reveals for the first time that soybeans GmSMS6 The biological functions of genes in controlling soybean grain quality traits provide new germplasm resources for genetic breeding work, offer new materials for soybean variety selection, and play a positive role in improving soybean varieties. Attached Figure Description

[0026] Figure 1 Showing GmSMS6 The gene structure diagram.

[0027] Figure 2 Showing GmSMS6 PCR detection results of transgenic material (using the Bar gene as the detection marker). Where M is a 2000+ DNA marker; 1-3 are WT; 4-5 are... GmSMS6 Mutant plant.

[0028] Figure 3 Showing GmSMS6 A schematic diagram of the acquisition of gene-edited materials and sequencing results.

[0029] Figure 4 Showing GmSMS6 Results of protein and oil content determination of mutant under Zp661 background. Detailed Implementation

[0030] 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.

[0031] 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.

[0032] The following examples used SPSS 11.5 statistical software to process the data. The experimental results are expressed as mean ± standard deviation. One-way ANOVA was used. P < 0.05 (*) indicates a significant difference, P < 0.01 (**) indicates a highly significant difference, and P < 0.001 (***) indicates a highly significant difference.

[0033] This invention uses Zhongpin661 (Zp661) as the acceptor material.

[0034] Zhongpin 661 is a regular commercial product.

[0035] Reference for strain EHA105: Komari T, Halperin W, Nester EW. Physical and functional map of supervirulent Agrobacterium tumefaciens tumor-inducing plasmid pTiBo542. J Bacteriol. 1986 Apr;166(1):88-94. doi: 10.1128 / jb.166.1.88-94.1986. PMID: 3957875; PMCID: PMC214561.

[0036] The aforementioned biological materials are available to the public from the Institute of Botany, Chinese Academy of Sciences. These biological materials are intended solely for repeating the relevant experiments of this invention and may not be used for any other purpose.

[0037] Example 1: Construction of Gene Editing Vector soybeans GmSMS6 The gene sequence was obtained from phytozome (https: / / phytozome-next.jgi.doe.gov / ). Figure 1Using targetDesign (http: / / skl.scau.edu.cn / targetdesign / ) GmSMS6 The design of sgRNA target sequences yielded two sgRNAs: sgRNA1: 5'-GTCGCCTACAAGAACGTAAT-3' (corresponding to positions 325-344 in sequence 1 and positions 160-179 in sequence 3); and sgRNA2: 5'-CAAGCTCCTCGACACGCGCC-3' (corresponding to positions 489-508 in sequence 1 and positions 324-343 in sequence 3), both located in... GmSMS6 On the first exon. Synthesize the corresponding forward and reverse primers for the sgRNA generated by the CRISPR-P 2.0 website.

[0038] soybeans GmSMS6 The genome sequence (SEQ ID NO.1, 2441bp) is as follows:

[0039] In SEQ ID NO.1, positions 1-165 are 5'UTR, positions 2003-2441 are 3'UTR; positions 166-654, 1336-1458, 1722-1838, and 1943-2002 are exons, and the rest are introns.

[0040] soybeans GmSMS6 The amino acid sequence of the protein GmSMS6 encoded by the gene is sequence 2 (SEQ ID NO. 2, 262aa) in the sequence listing, as follows: MAASAPTPREEFVYMAKLAEQAERYEEMVEFMEKVSASAESEELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEESRGNEDHVAVIRDYRSKIEAELSNICDGILKLLDTRLVPSAASGDSKVFYLKM KGDYHRYLAEFKTGADRKEAAESTLSAYKAAQDIANTELPPTHPIRLGLALNFSVFYYEILNSPDRACSLAKQAFDEAIAELDTLGEESYKDSTLIMQLLRDNLTLWTSDMQDDGADEIKEAAPKGDGEQN.

[0041] soybeans GmSMS6 The nucleic acid sequence (SEQ ID NO.3, 789bp) of the gene's coding sequence (CDS) is sequence 3 in the sequence listing, as follows: 5'--3'.

[0042] Build as follows GmSMS6 Gene knockout vector: The upstream primer for target 1 is B1-F:5'- GGAT TGTCGCCTACAAGAACGTAAT-3' (underscore base for forming sticky ends), downstream primer is B1-R: 5'- AAACATTACGTTCTTGTAGGCGACA-3' (underlined base for forming sticky ends), upstream primer B2-F for target 2: 5'- GGAT TGGCGCGTGTCGAGGAGCTTG-3', downstream primer is B2-R: 5'- AAAC CAAGCTCCTCGACACGCGCCA-3'.

[0043] B1-F and B1-R were annealed to form the annealed product B1-FR (a double-stranded DNA fragment with sticky ends). pGES201 was digested with BsaI to obtain a linearized vector of pGES201. B1-FR was then ligated into the linearized vector of pGES201 to obtain a vector expressing Cas9 and sgRNA targeting sgRNA1. GmSMS6 Gene knockout vector pCRISPR-Cas9-sgRNA1- GmSMS6 .

[0044] B2-F and B2-R were annealed to form the annealed product B2-FR (a double-stranded DNA fragment with sticky ends). pGES201 was digested with BsaI to obtain a linearized vector of pGES201. B2-FR was then ligated into the linearized vector of pGES201 to obtain a vector expressing Cas9 and sgRNA targeting sgRNA2. GmSMS6 Gene knockout vector pCRISPR-Cas9-sgRNA2- GmSMS6 .

[0045] pCRISPR-Cas9-sgRNA1- GmSMS6 Recombinant Agrobacterium tumefaciens EHA105 was obtained by introducing it into competent cells. GmSMS6 pCRISPR-Cas9-sgRNA2- GmSMS6 Recombinant Agrobacterium tumefaciens EHA105 was obtained by introducing it into competent cells. GmSMS6 .

[0046] Example 2 Agrobacterium-mediated transformation of soybean cotyledonary nodes Recombinant Agrobacterium EHA105 / pCRISPR-Cas9-sgRNA1- GmSMS6 and recombinant Agrobacterium EHA105 / pCRISPR-Cas9-sgRNA2- GmSMS6 Soybean cotyledonary nodes were infected with a 1:1 mixture of CFU (colony-forming units) for soybean genetic transformation. The specific procedure included: (1) Soybean seed cleaning and disinfection: Soybean Zp661 is used as the recipient material. Select seeds that are plump, uniform in size and free of disease spots on the surface. Disinfect overnight by reacting sodium hypochlorite with concentrated hydrochloric acid to release chlorine gas.

[0047] (2) Explant preparation and bacterial inoculation: The treated soybean seeds were placed in a clean conical flask and soaked overnight in sterile water. In a laminar flow hood, the seed coat was cut along the hilum with a scalpel to separate the embryo into two halves. The cotyledon explants were placed in a bacterial solution resuspended to an OD value of 0.6-0.8 and ultrasonically treated for 3 minutes. The conical flask containing the explants was then placed in a vacuum pump and treated for 10 minutes (0.6 Pa). Finally, it was placed in a shaker and cultured at 23°C and 110 rpm for 40 minutes.

[0048] (3) Co-culture: After discarding the bacterial culture, spread the explants evenly on clean filter paper, blow dry, and spread them evenly on the co-culture medium (B5 medium + 3% sucrose + 0.8% Agar + 3mM MES + 0.25mg / L gibberellin (GA3) + 1.67mg / L benzylaminopurine (6-BA) + 580mg / L cysteine ​​+ 154.2mg / L dithiothreitol + 200µM eugenol (AS), pH 5.4), place them in an incubator, and culture in the dark at 23℃ for 3 days. Then transfer them to a tissue culture room at 23-25℃, with a photoperiod of 16 h light / 8 h dark, and culture for 1 day.

[0049] (4) Recovery culture: The hypocotyl of the explant was partially removed, leaving 3-4 mm, and inserted obliquely into the recovery culture medium (MS medium + 3% sucrose + 3mM MES + 0.8% Agar + 1.67 mg / L benzylaminopurine (6-BA) + 100 mg / L cephalosporin (Cef), pH 5.7), and sealed. It was placed in the tissue culture room with a photoperiod of 16 h light / 8 h darkness for 7 days.

[0050] (5) Bud induction and selection culture: Excessively long hypocotyls were removed, and explants were obliquely inserted into the selection medium (MS medium + 3% sucrose + 3mM MES + 0.8% Agar + 1.67mg / L benzylaminopurine (6-BA) + 5mg / L glutfosinate + 100mg / L cephalosporin (Cef), pH 5.7). The explants were placed in the tissue culture room with a photoperiod of 16 h light / 8 h dark for 21 days.

[0051] (6) Bud elongation culture: Remove the cotyledons of the explant, retaining the clustered buds and base, and make a new wound at the base. Place the explant on bud induction medium (MS medium + 3% sucrose + 3mM MES + 0.8% Agar + 50mg / L asparagine + 50mg / L glutamine + 0.1mg / L indoleacetic acid (IAA) + 0.5mg / L gibberellin (GA3) + 1mg / L zeatin + 5mg / L glutfosinate + 500mg / L termethin (Tim) + 100mg / L cephalosporin (Cef), pH 5.7), and place it in a tissue culture room at 23-25℃ with a photoperiod of 16 h light / 8 h darkness for 2-8 weeks. During this period, change the medium or induce root growth according to the growth status of the explant.

[0052] (7) Root induction culture: When the shoots elongate to 3-5 cm, cut the elongated shoots from the base of the tissue, soak the cut end in auxin solution for 2 minutes, and then transfer them into rooting medium (MS medium + 3% sucrose + 3mM MES + 0.8% Agar + 1mg / LIBA, pH 5.7). Place them in the tissue culture room for root induction, which generally takes about 2 weeks.

[0053] (8) Seedling transplanting: Transplant the seedlings into the soil, cover them with a thin film and place them in the light. After the plants have grown normally for 3-5 days, remove the film to obtain T0 generation plants. Harvest T0 generation seeds, self-pollinate the T0 generation to obtain T1 generation seeds, plant the T1 generation seeds to obtain T1 generation seedlings and perform the following tests.

[0054] Example 3: Identification of transgenic plants To identify transgenic positive plants, DNA was extracted from leaves of tissue culture seedlings and subjected to PCR detection. First, the bar gene was detected, and the results are as follows: Figure 2 As shown, M stands for Marker, 1-3 are WT (Zp661) strains, denoted as WT_1, WT_2, and WT_3 respectively; 4 is gene-edited strain Z1, and 5 is gene-edited strain Z2.

[0055] Using genomic DNA from gene-edited lines Z1 and Z2 as templates, PCR amplification was performed using upstream primer F: 5'-CAATCCCAGCCTTTCGGTT-3' and downstream primer R: 5'-TCAATTTTCAACATCCATCGCTT-3'. The PCR products were then subjected to Sanger sequencing to identify the mutation status of the target sites in the gene-edited lines.

[0056] Target 1 sgRNA1: 5'-GTCGCCTACAAGAACGTAAT-3' (corresponding to positions 325-344 in sequence 1, and positions 160-179 in sequence 3); Target 2 sgRNA2: 5'-CAAGCTCCTCGACACGCGCC-3' (corresponding to positions 489-508 in sequence 1, and positions 324-343 in sequence 3). Z1 and Z2 are both... GmSMS6 Gene-edited homozygous plants.

[0057] Sequencing results were compared with wild-type Zp661, with edits as follows: Figure 3 As shown. In Z1, compared with soybean Zp661, the genome of Z1 has... GmSMS6 The corresponding region of the gene underwent the following changes: a deletion between positions 342 and 344 in sequence 1 (corresponding to positions 177 and 179 in sequence 3), and an insertion of an adenine ribonucleotide between positions 491 and 492 (corresponding to positions 326 and 327 in sequence 3), thereby altering the region of the soybean Zp661 genome. GmSMS6 Gene knockout; in Z2, compared to soybean Zp661, Z2... GmSMS6 The region corresponding to the gene underwent the following changes: between positions 336 and 340 in sequence 1 (corresponding to positions 171 and 175 in sequence 3), thereby altering the region of the soybean Zp661 genome. GmSMS6 Gene knockout.

[0058] The protein and oil content of mature seeds from wild-type soybean Zp661 and gene-edited plants Z1 and Z2 were measured using a near-infrared spectroscopy analyzer (Bruker, MPA Near-infrared Spectrum System, Germany). Figure 4 It was found that, compared with the wild-type Zp661, the gene-edited plants showed increased Z1 and Z2 protein contents of 11.46% and 12.70%, respectively, and decreased oil contents of 9.99% and 10.19%, respectively. These results indicate... GmSMS6 It can regulate the content of soybean oil and protein.

[0059] 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.

[0060] This application is supported by the National Science and Technology Major Project for Agricultural Biotechnology Breeding (2023ZD0406904).

Claims

1. A method for regulating plant seed traits, characterized in that, The method includes regulating the seed traits of a target plant by gene knockout, wherein the gene knockout is the knockout of the gene encoding the target protein in the target plant, and the target plant contains the gene encoding the target protein. The target protein is GmSMS6, and is any one of the following: A1) The amino acid sequence is that of the protein shown in sequence 2. A2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein described in A1), which has more than 80% identity with the protein shown in A1) and can regulate plant grain traits. A3) A fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of A1) or A2).

2. A method for cultivating plants with altered grain traits, comprising obtaining a target plant with altered grain traits by gene knockout, wherein the target plant contains the gene encoding the protein of claim 1.

3. The method according to claim 1 or 2, characterized in that, The protein is derived from soybeans.

4. The method as described in claims 1-3, characterized in that, The genome of the target plant contains a DNA molecule whose nucleotide sequence is sequence 1 in the sequence listing, and the gene knockout includes performing any of the following operations on the genome of the target plant: B1) Delete nucleotides from position 342 to 344 of sequence 1 in the sequence listing of the target plant, and insert an adenine ribonucleotide between positions 491 and 492; B2) Delete nucleotides 336-340 from sequence 1 in the sequence listing of the target plant.

5. The method according to any one of claims 1-4, characterized in that, The plant seed characteristics are to increase the protein content of plant seeds and / or decrease the oil content of plant seeds.

6. The method according to any one of claims 1-5, characterized in that, The plant is any one of the following: C1) Dicotyledons, C2) Plants of the Rosales order, C3) Leguminosae (family legumes) C4) Plants of the genus *Glycine*. C5) Soybeans.

7. The application of biomaterials, characterized in that, The application is any one of the following: D1) The application of the biomaterials described herein in regulating plant seed traits and / or in preparing products that regulate the desired plant seed traits. D2) The application of the biomaterials described therein in plant breeding and / or the preparation of plant breeding products; The biological material is a protein, a substance that regulates the expression of the gene encoding the protein, or a substance that regulates the content of the protein. The protein is GmSMS6, and is any one of the following: The amino acid sequence of E1 is the protein shown in sequence 2. E2) A protein obtained by substituting and / or deleting and / or adding amino acid residues of the protein described in E1), which has more than 80% identity with the protein shown in E1) and can regulate plant grain traits. E3) is a fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of E1) or E2).

8. The application according to claim 7, characterized in that, The substance is any one of the following: F1) Nucleic acid molecules that inhibit, reduce, or downregulate the expression of the gene encoding the protein described in claim 7. F2) expresses the gene encoding the nucleic acid molecule described in F1). F3) contains the expression cassette of the gene described in F2). F4) a recombinant vector containing the gene described in F2), or a recombinant vector containing the expression cassette described in F3). F5) Recombinant microorganisms containing the gene described in F2), or recombinant microorganisms containing the expression cassette described in F3), or recombinant microorganisms containing the recombinant vector described in F4). F6) A transgenic plant cell line containing the gene described in F2), or a transgenic plant cell line containing the expression cassette described in F3), or a transgenic plant cell line containing the recombinant vector described in F4). F7) Transgenic plant tissue containing the gene described in F2), or transgenic plant tissue containing the expression cassette described in F3), or transgenic plant tissue containing the recombinant vector described in F4), F8) A transgenic plant organ containing the gene described in F2), or a transgenic plant organ containing the expression cassette described in F3), or a transgenic plant organ containing the recombinant vector described in F4).

9. The application as described in claim 7 or 8, characterized in that, The regulation of plant grain traits involves increasing the protein content and / or decreasing the oil content of plant grains.

10. The application as described in any one of claims 7-9, characterized in that, The plant is any one of the following: G1) Dicotyledons, G2) Plants of the Rosales order, G3) Leguminosae (family legumes) G4) Soybean genus plants, G5) Soybeans.

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