GmTCP27 gene and application thereof in regulation and control of plant seed size

Abstract

The invention discloses a GmTCP27 gene and an application thereof in regulating and controlling the size of plant seeds, the expression quantity of a target gene GmTCP27 in soybeans is increased or the target gene GmTCP27 is subjected to overexpression, and a transgenic plant with larger seeds is cultivated; the CDS sequence of the gene GmTCP27 is as shown in SEQ ID NO: 1. According to the invention, a coding sequence of the GmTCP27 gene is constructed into an overexpressed binary vector, and overexpression of the GmTCP27 gene can promote soybean seed enlargement by utilizing a cotyledonary node stable transformation technology. Therefore, overexpression of the GmTCP27 can achieve the purpose of promoting plant seed enlargement, and the gene can be fully utilized for soybean variety improvement subsequently, which is beneficial to improvement of soybean yield.

Description

Technical Field

[0001] The present invention relates to the field of genetic engineering technology, in particular to the GmTCP27 gene and its application in regulating plant seed size. Background Art

[0002] Soybean (Glycine max) originated in China and has a domestication history of more than five thousand years. It is currently the leguminous crop with the largest planting area in China. Relying on molecular design breeding technology to create new varieties with high yield, stress resistance and wide adaptability, ultimately promoting a substantial increase in the yield per mu of soybean and providing an important guarantee for national food security.

[0003] Seed size is an important agronomic trait for improving soybean yield. Current research shows that soybean seed size can be regulated through the gibberellin pathway, brassinolide pathway, cytokinin pathway, and transcription factor regulation, etc. Soybean seed size includes seed length (SL), seed width (SW), and seed thickness (ST), which are key appearance parameters affecting seed weight, yield, and grain use. However, the molecular mechanisms of soybean seed length, seed width, and seed thickness traits are not yet clear, and relatively few important genetic loci (QTL) and genes available for molecular breeding.

[0004] There is a certain homology of genes among related species, and the functions of many genes are relatively conserved among different species. Currently, the research on seed size in Arabidopsis thaliana and rice is relatively thorough. Using the method of homologous gene cloning to mine genes regulating seed size in soybean is one of the effective methods.

[0005] TCP (TEOSINTE BRANCHED1 / CYCLOIDEA / PROLIFERATING CELL FACTOR) is a class of plant-specific transcription factor families, which play a crucial role in plant growth and development and stress responses. The TCP family in soybean includes a total of 55 members, which can be divided into Class I and Class II, and Class II can be further divided into CIN and CYC / TB1. Some studies have shown that members GmTCP13 and GmTCP40 in the soybean TCP family can promote flowering, GmTCP9-like can improve salt tolerance, GmTCP19-L can respond to Phytophthora sojae, GmTCP14 and GmTCP29 can regulate soybean leaf shape, and GmTCP14 can also regulate the biosynthesis of soybean isoflavones. In Arabidopsis thaliana, it has been reported that TCP14 and TCP15 can regulate the size of plant organs including seeds. In rice, studies have shown that inhibiting OsTCP4 can increase the tiller number and reduce the seed length. Currently, in soybean, the functions and action mechanisms of TCP family members in regulating seed size have not been reported. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide the GmTCP27 gene and its application in regulating plant seed size.

[0007] To solve the above technical problems, the technical solutions adopted by the present invention are as follows.

[0008] Use of the GmTCP27 protein in the following 1)-3):

[0009] 1) Regulating plant seed size;

[0010] 2) Cultivating transgenic plants with larger seeds;

[0011] 3) Cultivating transgenic plants with smaller seed size;

[0012] The GmTCP27 protein is a1) or a2) or a3) or a4):

[0013] a1) A protein with the amino acid sequence shown in SEQ ID NO: 2;

[0014] a2) A fusion protein obtained by connecting a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO: 2;

[0015] a3) A protein related to plant seed size obtained by substituting and / or deleting and / or adding one or several amino acid residues to the amino acid sequence shown in SEQ ID NO: 2;

[0016] a4) A protein having 90% identity with the amino acid sequence shown in SEQ ID NO: 2, derived from soybean and related to plant seed size.

[0017] Use of a biological material related to the GmTCP27 protein in the following 1)-3):

[0018] 1) Regulating plant seed size;

[0019] 2) Cultivating transgenic plants with smaller seeds;

[0020] 3) Cultivating transgenic plants with larger seeds;

[0021] The biological material is any one of the following A1) to A8):

[0022] A1) A nucleic acid molecule encoding the GmTCP27 protein;

[0023] A2) An expression cassette containing the nucleic acid molecule described in A1);

[0024] A3) A recombinant vector containing the nucleic acid molecule described in A1);

[0025] A4) A recombinant vector containing the expression cassette described in A2);

[0026] A5) A recombinant microorganism containing the nucleic acid molecule described in A1);

[0027] A6) A recombinant microorganism containing the expression cassette described in A2);

[0028] A7) A recombinant microorganism containing the recombinant vector described in A3);

[0029] A8) A recombinant microorganism containing the recombinant vector described in A4).

[0030] As a preferred technical solution of the present invention, the nucleic acid molecule described in A1) is a gene shown in any of B1) or B2) or B3) or B4) as follows:

[0031] B1) The CDS molecule shown in SEQ ID NO: 1;

[0032] B2) The genomic DNA molecule corresponding to the CDS molecule shown in SEQ ID NO: 1;

[0033] B3) A CDS molecule or genomic DNA molecule having 75% or more identity with the nucleotide sequence defined by B1) or B2) and encoding the GmTCP27 protein described in claim 1;

[0034] B4) A CDS molecule or genomic DNA molecule that hybridizes with the nucleotide sequence defined by B1) or B2) or B3) under stringent conditions and encodes the GmTCP27 protein described in claim 1.

[0035] Use of the substance shown by m1 or m2 in increasing the size of plant seeds or cultivating transgenic plants with larger seeds;

[0036] m1, a substance that overexpresses the activity or content of the GmTCP27 protein in plants;

[0037] m2, a substance that overexpresses the nucleic acid encoding the GmTCP27 protein in plants or a substance of the nucleic acid encoding the GmTCP27 protein in plants;

[0038] The GmTCP27 protein is any of a1) or a2) or a3) or a4):

[0039] a1) A protein with an amino acid sequence shown in SEQ ID NO: 2;

[0040] a2) A fusion protein obtained by connecting a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO: 2;

[0041] a3) A protein related to plant seed size obtained by substituting and / or deleting and / or adding one or more amino acid residues to the amino acid sequence shown in SEQ ID NO: 2;

[0042] a4) A protein having 90% identity to the amino acid sequence shown in the sequence, derived from soybean and related to plant seed size.

[0043] A method for cultivating a transgenic plant with larger seeds, comprising the following steps: increasing the content and / or activity of the GmTCP27 protein described in claim 1 in a recipient plant to obtain a transgenic plant; the seeds of the transgenic plant are larger than those of the recipient plant.

[0044] As a preferred technical solution of the present invention, the method for increasing the content and / or activity of the GmTCP27 protein described in claim 1 in the recipient plant is to overexpress the GmTCP27 protein in the recipient plant;

[0045] and / or, the method of overexpression is to introduce the coding gene of the GmTCP27 protein into the recipient plant.

[0046] A method for cultivating a transgenic plant with smaller seeds, comprising the following steps: decreasing the content and / or activity of the GmTCP27 protein described in claim 1 in a recipient plant to obtain a transgenic plant; the seeds of the transgenic plant are smaller than those of the recipient plant.

[0047] As a preferred technical solution of the present invention, the method for decreasing the content and / or activity of the GmTCP27 protein described in claim 1 in the recipient plant is achieved by knocking out or inhibiting or silencing the coding gene of the GmTCP27 protein described in claim 1 in the recipient plant using RNAi technology;

[0048] Furthermore, the substance for silencing the coding gene of the GmTCP27 protein in the recipient plant is a nucleic acid molecule that interferes with the expression of the coding gene of the GmTCP27 protein in the recipient plant.

[0049] A method for increasing plant yield, by overexpressing the expression of the GmTCP27 gene in the plant to increase the size of plant seeds, thereby increasing the yield of the corresponding plant.

[0050] As a preferred technical solution of the present invention, the above-mentioned plant is soybean.

[0051] The beneficial effects of adopting the above technical solution are as follows: In the present invention, the coding sequence of the GmTCP27 gene is constructed into an overexpression binary vector, and it is found by using the stable cotyledon node transformation technology that its overexpression can promote the enlargement of soybean seeds. Therefore, overexpressing the GmTCP27 gene can achieve the purpose of promoting the enlargement of plant seeds, and this gene can be fully utilized for the improvement of soybean varieties in the future, which helps to increase the yield of soybeans. Brief Description of the Drawings

[0052] Figure 1 It is a diagram for analyzing the tissue expression pattern of soybean GmTCP27, detecting the expression level of GmTCP27 in roots, hypocotyls, cotyledons, the first trifoliate leaf, apical meristem, flowers and green pods by using RT-qPCR technology;

[0053] Figure 2 It is a diagram for constructing the pUbi:GmTCP27-3FLAG overexpression vector. In the figure, A is a schematic diagram of the pUbi:GmTCP27-3FLAG vector; B is the digestion of the pUBI:GmTCP27-3FLAG vector using KpnⅠ and SacⅠ, and the target band is 800bp. The "+" represents adding restriction endonucleases, and the "-" represents not adding restriction endonucleases;

[0054] Figure 3 It is a diagram of the WT wild-type control line and the pUbi:GmTCP27-3FLAG overexpression line in soybeans;

[0055] Figure 4 It is a diagram for observing the seed size phenotype of the WT wild-type control line and the pUbi:GmTCP27-3FLAG overexpression line in soybeans. In the figure, A is the phenotype of the grain length, grain width and grain thickness of the mature dry seeds of the WT wild-type control and the pUbi:GmTCP27-3FLAG overexpression lines (23-2, 5-1), and the scale bar is 3cm; B is the statistical chart of the grain length, grain width, grain thickness and 100-seed weight of the pUbi:GmTCP27-3FLAG overexpression lines (23-2, 5-1). t-test, **P<0.01, ***P<0.001 and ****p<0.0001 respectively indicate very significant difference, extremely significant difference and extremely extremely significant difference;

[0056] Figure 5 It is a diagram for constructing the p35S:GmTCP27-GFP overexpression vector. In the figure, A is a schematic diagram of the p35S:GmTCP27-GFP vector; B is the digestion of the p35S:GmTCP27-GFP vector using XbaⅠ and SacⅠ, and the target band is 1485bp. The "+" represents adding restriction endonucleases, and the "-" represents not adding restriction endonucleases;

[0057] Figure 6 Full resistance screening diagrams and transgenic line diagrams of the Col wild-type control line and the p35S:GmTCP27-GFP overexpression line in Arabidopsis thaliana;

[0058] Figure 7 Phenotype observation diagrams of seed size of the Col wild-type control line and the p35S:GmTCP27-GFP overexpression line in Arabidopsis thaliana. In the figure, A shows the phenotypes of grain length, grain width, and grain thickness of mature dry seeds of the Col wild-type control and the p35S:GmTCP27-GFP overexpression lines (2-4 and 5-7), and the scale bar is 20 μm; B shows the statistical graphs of grain length, grain width, grain thickness, and 1000-grain weight of the Col wild-type control and the p35S:GmTCP27-GFP overexpression lines (2-4 and 5-7). t-test, ****p<0.0001, indicating extremely significant differences. Detailed implementation manners

[0059] The following examples illustrate the present invention in detail. All kinds of raw materials and various equipment used in the present invention are conventional commercially available products and can be directly obtained through market purchase. The experimental methods used in the following examples are all conventional methods unless otherwise specified.

[0060] It should be understood that when used in the specification of the present application and the appended claims, the term "comprising" indicates the presence of the described features, wholes, steps, operations, elements, and / or components, but does not exclude the presence or addition of one or more other features, wholes, steps, operations, elements, components, and / or their combinations.

[0061] It should also be understood that the term "and / or" used in the specification of the present application and the appended claims refers to any combination and all possible combinations of one or more of the associated listed items, and includes these combinations.

[0062] Reference to "one embodiment" or "some embodiments" etc. described in the specification of the present application means that specific features, structures, or characteristics described in connection with that embodiment are included in one or more embodiments of the present application. Thus, statements such as "in one embodiment", "in some embodiments", "in other some embodiments", "in still other embodiments" etc. that appear in different places in this specification are not necessarily all referring to the same embodiment, but mean "one or more but not all embodiments", unless otherwise specifically emphasized in other ways. The terms "comprising", "including", "having" and their variants all mean "including but not limited to", unless otherwise specifically emphasized in other ways.

[0063] In addition, in the description of the specification and the appended claims of the present application, the terms "first", "second", "third", etc. are only used for distinguishing descriptions and cannot be construed as indicating or implying relative importance.

[0064] Next, specific embodiments of the present invention will be combined to clearly and completely describe the technical solutions of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0065] The soybean GmTCP27 gene (Glyma.10G285900) provided by the present invention encodes a transcription factor containing a TCP domain. Its CDS sequence is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. The expression level of this gene is relatively high in roots.

[0066] Example 1. Detection of the tissue expression pattern of GmTCP27 in soybeans

[0067] Primer design: Select the actin GmActin11 in soybeans as the internal reference gene, and design quantitative primers on the CDS sequences of GmActin11 and GmTCP27 so that the size of the PCR product is between 150-200 bp. The specific sequences are as follows:

[0068] Table 1 Primer pair sequence table

[0069]

[0070] Obtaining tissue materials: Take partial tissue materials of the soybean Williams 82 plants that are two weeks old, including roots, hypocotyls, cotyledons, and leaves; when they grow to about 1 cm in size with green pods, take partial tissue materials of the Williams 82 plants, including flowers, apical meristems, and green pods.

[0071] After extracting the RNA of each part of the tissue using the TRIZOL method, perform a mixed reverse transcription system according to Table 2.

[0072] Table 2 Reverse transcription system

[0073]

[0074] After mixing the above system, place it in a 42°C metal bath for 2 minutes, and then add 4 μL of 5×HiScriptⅡ qRTSuperMixⅡ and mix evenly. Perform reverse transcription according to the procedure in Table 3.

[0075] Table 3 Reverse transcription procedure

[0076]

[0077] After the reverse transcription was completed, 180 μL of RNase-free ddH2O was added.

[0078] Quantitative fluorescence PCR was performed using the quantitative fluorescence PCR enzyme from Novoprotein Scientific Inc. according to the system shown in Table 4.

[0079] Table 4 Quantitative fluorescence PCR system

[0080]

[0081] Table 5 Quantitative fluorescence PCR program

[0082]

[0083] Using GmActin11 as the internal reference gene, the expression levels of the GmTCP27 gene in various tissues were calculated. As Figure 1 shown, the expression level of the GmTCP27 gene was the highest in the roots, followed by the leaves, flowers, apical meristems, cotyledons, and finally the lowest in the hypocotyls and green pods.

[0084] Example 2. Cloning of the GmTCP27 gene

[0085] Primer design: For the upstream primer required for amplifying the target gene, a 15-bp upstream vector sequence and a 6-bp recognition sequence for the target restriction enzyme Kpn I need to be added at the 5' end of the ATG. For the downstream primer, a 15-bp downstream vector sequence and a 6-bp recognition sequence for the target restriction enzyme Kpn I need to be added at the 5' end of the original downstream primer sequence. The specific primer sequences are as follows:

[0086] Table 6 Primer pair sequence list

[0087]

[0088] Amplification was performed using the high-fidelity KOD FX polymerase according to the system shown in Table 7.

[0089] Table 7 PCR amplification system

[0090]

[0091] Table 8 PCR amplification program

[0092]

[0093] After the amplification was completed, the products were recovered using an agarose gel product recovery kit. The specific steps refer to the kit instruction manual of Jierui Biotechnology Co., Ltd.

[0094] Example 3. Obtaining of Arabidopsis overexpression transgenic lines

[0095] Using the pCAMBIA 1300 35S:GFP binary vector as a backbone, the vector was linearized with Xba I and BamH I restriction endonucleases, and then a one-step cloning ligation reaction was carried out using the system shown in Table 9:

[0096] Table 9 Cloning ligation reaction system

[0097]

[0098] Note: The most suitable amount of linearized vector added (b) = [0.02 × number of base pairs of the cloning vector] ng;

[0099] The most suitable amount of insert fragment used (a) = [0.04 × number of base pairs of the insert fragment] ng.

[0100] The above ligation system was placed in a 37°C metal bath. After reacting for 15 min, it was transformed into DH5α. After digestion identification (such as Figure 5 ), Sanger sequencing was carried out to obtain the p35S:GmTCP27-GFP plant expression vector with correct sequencing. Through the floral dip method of Arabidopsis thaliana, the p35S:GmTCP27-GFP overexpression transgenic Arabidopsis thaliana lines were obtained (such as Figure 6 ).

[0101] Example 4 Observation of the seed size phenotype of Arabidopsis thaliana GmTCP27 heterologous overexpression lines

[0102] The seeds of the Col wild-type control to be detected and the p35S:GmTCP27-GFP overexpression lines were scattered as evenly as possible on white paper. The seeds were simply arranged with the needle of a syringe, and then collected under a stereomicroscope (LEICA M250 FA). Under the same setting conditions, a ruler was photographed as a scale. After collection, the area of the seeds was measured using Image J software, and the measurement results were statistically analyzed using Prism software. The results are shown in Table 10, Table 11, Table 12 and Figure 7 .

[0103] Table 10 Statistical table of seed length, width and thickness of Col wild-type control lines and p35S:GmTCP27-GFP overexpression lines in Arabidopsis thaliana

[0104]

[0105]

[0106] Statistical results of 1000-seed weight of Col wild-type control lines and p35S:GmTCP27-GFP overexpression lines in Arabidopsis thaliana

[0107]

[0108] Statistical analysis results of seed length, seed width, seed thickness, and 1000-seed weight of Col wild-type control lines and p35S:GmTCP27-GFP overexpression lines in Arabidopsis thaliana

[0109]

[0110]

[0111] As shown in Tables 10, 11, and 12 and Figure 7 it can be seen that the differences in seed length, seed width, seed thickness, and 1000-seed weight between the Col wild-type control lines and the p35S:GmTCP27-GFP overexpression lines all reached a significant level (P<0.05). Among them, the seed length, seed width, seed thickness, and 1000-seed weight of the p35S:GmTCP27-GFP overexpression lines were greater than those of the Col wild-type control lines.

[0112] In this invention, by constructing the p35S:GmTCP27-GFP vector and overexpressing it in Arabidopsis thaliana, and statistically analyzing the seed size phenotypes of the overexpression lines, the results fully indicate that the overexpression of the GmTCP27 gene has the effect of promoting the seed size of Arabidopsis thaliana. According to the above content, the GmTCP27 gene has application value in the research and improvement of Arabidopsis thaliana seed size.

[0113] Example 5. Obtaining of soybean overexpression transgenic lines

[0114] Using the pUbi:3FLAG binary vector as the backbone, linearize the vector with Kpn I restriction endonuclease, and then perform a one-step cloning (One Step Cloning) ligation reaction using the system shown in Table 9 above. Incubate the above ligation system in a 37°C metal bath, transform DH5α after reacting for 15 min, and after restriction enzyme digestion identification (such as Figure 2 ), perform Sanger sequencing to obtain the pUbi:GmTCP27-3FLAG plant expression vector with correct sequencing, and obtain the pUbi:TCP27-3FLAG overexpression transgenic soybean lines through the stable transformation method of cotyledon nodes (such as Figure 3 ).

[0115] Example 6. Observation of seed size phenotypes of soybean pUbi:GmTCP27-3FLAG overexpression lines

[0116] Randomly weigh 100 soybean seeds each from the mature WT wild-type control strain to be tested and the pUbi:GmTCP27-3FLAG overexpression strain, and then place them on a thousandth scale to count the 100-seed weight, repeating three times. Use a vernier caliper with a precision of 0.1 mm to measure the seed length, width, and thickness of the soybean seeds. The measurement results are statistically analyzed using Prism software. The results are shown in Table 13, Table 14, Table 15 and Figure 4 as follows.

[0117] Table 13 Statistical table of seed length, width, and thickness of soybean WT wild-type control strain and pUbi:GmTCP27-3FLAG overexpression strain

[0118]

[0119]

[0120] Table 14 Statistical results of 100-seed weight of seeds of soybean WT wild-type control strain and pUbi:GmTCP27-3FLAG overexpression strain

[0121]

[0122] Table 15 Statistical analysis results of seed length, width, thickness, and 100-seed weight of seeds of soybean WT wild-type control strain and pUbi:GmTCP27-3FLAG overexpression strain

[0123]

[0124] As can be seen from Table 13, Table 14, Table 15 and Figure 4 it can be seen that the differences in seed length, width, thickness, and 100-seed weight between the WT wild-type control strain and the pUbi:GmTCP27-3FLAG overexpression strain all reached a significant level (P<0.05). Among them, the seed length, width, thickness, and 100-seed weight of the pUbi:GmTCP27-3FLAG overexpression strains 23-2 and 5-1 were all greater than those of the WT wild-type control strain.

[0125] In this invention, by constructing the pUbi:GmTCP27-3FLAG vector, overexpression was carried out in soybeans, and the 100-seed weight of the seeds of the overexpression strains was counted. The results fully showed that overexpression of the GmTCP27 gene has the effect of promoting the size of soybean seeds. According to the above content, the GmTCP27 gene has application value in the research and improvement of soybean seed size.

[0126] In summary, in the above embodiments, the coding sequence of the GmTCP27 gene was constructed into an overexpression binary vector, and it was found by using the stable cotyledon node transformation technology that its overexpression could promote the enlargement of soybean seeds. Therefore, overexpressing GmTCP27 can achieve the purpose of promoting the enlargement of plant seeds, and this gene can be fully utilized for the improvement of soybean varieties in the future, which helps to increase the yield of soybeans.

[0127] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these examples without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

[0128] In the above embodiments, the descriptions of the respective embodiments have their own emphases. For parts not detailed or recorded in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0129] The above-described embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the same; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention, and should all be included in the protection scope of the present invention.

Claims

1. Use of the GmTCP27 protein in the following 1)-3): 1) Regulating plant seed size; 2) Cultivating transgenic plants with larger seeds; 3) Cultivating transgenic plants with smaller seed size; The GmTCP27 protein is one of a1) or a2) or a3) or a4): a1) A protein with the amino acid sequence shown in SEQ ID NO: 2; a2) A fusion protein obtained by linking a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO: 2; a3) A protein related to plant seed size obtained by substituting and / or deleting and / or adding one or several amino acid residues to the amino acid sequence shown in SEQ ID NO: 2; a4) A protein with 90% identity to the amino acid sequence shown in SEQ ID NO: 2, derived from soybean and related to plant seed size.

2. Use of a biological material related to the GmTCP27 protein in the following 1)-3): 1) Regulating plant seed size; 2) Cultivating transgenic plants with smaller seeds; 3) Cultivating transgenic plants with larger seeds; The biological material is any one of the following A1) to A8): A1) A nucleic acid molecule encoding the GmTCP27 protein; A2) An expression cassette containing the nucleic acid molecule described in A1); A3) A recombinant vector containing the nucleic acid molecule described in A1); A4) A recombinant vector containing the expression cassette described in A2); A5) A recombinant microorganism containing the nucleic acid molecule described in A1); A6) A recombinant microorganism containing the expression cassette described in A2); A7) A recombinant microorganism containing the recombinant vector described in A3); A8) A recombinant microorganism containing the recombinant vector described in A4).

3. The application according to claim 2, wherein: The nucleic acid molecule described in A1) is a gene shown in any one of B1) or B2) or B3) or B4) below: B1) The CDS molecule shown in SEQ ID NO: 1; B2) The genomic DNA molecule corresponding to the CDS molecule shown in SEQ ID NO: 1; B3) A CDS molecule or genomic DNA molecule having 75% or more identity to the nucleotide sequence defined by B1) or B2) and encoding the GmTCP27 protein described in claim 1; B4) A CDS molecule or genomic DNA molecule that hybridizes with the nucleotide sequence defined by B1) or B2) or B3) under stringent conditions and encodes the GmTCP27 protein described in claim 1.

4. Use of the substance shown in m1 or m2 in increasing plant seed size or cultivating transgenic plants with larger seeds; m1, a substance that overexpresses the activity or content of the GmTCP27 protein in plants; m2, a substance that overexpresses the expression of the nucleic acid encoding the GmTCP27 protein in plants or a substance that overexpresses the nucleic acid encoding the GmTCP27 protein in plants; The GmTCP27 protein is one of a1) or a2) or a3) or a4): a1) A protein with the amino acid sequence shown in SEQ ID NO: 2; a2) A fusion protein obtained by linking a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID NO: 2; a3) A protein related to plant seed size obtained by substituting and / or deleting and / or adding one or more amino acid residues to the amino acid sequence shown in SEQ ID NO: 2; a4) A protein having 90% identity with the amino acid sequence shown in the sequence, derived from soybean and related to plant seed size.

5. A method for cultivating a transgenic plant with larger seeds, characterized in that: It includes the following steps: increasing the content and / or activity of the GmTCP27 protein described in claim 1 in a recipient plant to obtain a transgenic plant; the seeds of the transgenic plant are larger than those of the recipient plant.

6. The method according to claim 5, wherein: The method for increasing the content and / or activity of the GmTCP27 protein described in claim 1 in the recipient plant is overexpressing the GmTCP27 protein in the recipient plant; and / or, the method for overexpression is introducing the coding gene of the GmTCP27 protein into the recipient plant.

7. A method for cultivating a transgenic plant with smaller seeds, characterized in that: It includes the following steps: decreasing the content and / or activity of the GmTCP27 protein described in claim 1 in a recipient plant to obtain a transgenic plant; the seeds of the transgenic plant are smaller than those of the recipient plant.

8. The method according to claim 7, characterized in that: The method for decreasing the content and / or activity of the GmTCP27 protein described in claim 1 in the recipient plant is achieved by knocking out or inhibiting or silencing the coding gene of the GmTCP27 protein described in claim 1 in the recipient plant using RNAi technology; Furthermore, the substance for silencing the coding gene of the GmTCP27 protein in the recipient plant is a nucleic acid molecule that interferes with the expression of the coding gene of the GmTCP27 protein in the recipient plant.

9. A method for increasing plant yield, characterized in that: Overexpressing the expression of the GmTCP27 gene in the plant to increase the size of plant seeds, thereby increasing the yield of the corresponding plant.

10. According to any one of claims 1-4, the application, or the method according to claim 5 or 6, or the method according to claim 7 or 8, or the method according to claim 9, characterized in that: The plant is soybean.

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