Soybean GmMADS33 and its homologs and applications

Abstract

This invention discloses soybean GmMADS33, its homologous gene, and its applications. The amino acid sequence of GmMADS33 is shown in SEQ ID NO.3, and the nucleotide sequence of its encoding gene is shown in SEQ ID NO.1. Compared to the cultivated soybean variety Williams, gmmad33 Single protrusion and gmmads33 / gmmads34 Soybeans exhibited reduced pod width and projected area, as well as a decrease in 100-kernel weight; GmMADS33 Soybeans with -OE exhibit increased pod area, higher 100-seed weight, and increased number of pods, seeds, and yield per plant. Therefore, GmMADS33 These genes play an important role in regulating pod size, pod number, seed number, and seed weight, and are key regulatory genes for soybean pod size, seed weight, and seed number.

Description

Technical Field

[0001] This invention belongs to the field of plant molecular biology, specifically relating to soybean GmMADS33 and its homologous genes and their applications. Background Technology

[0002] Developing high-yield and high-quality soybean varieties is the core objective of the soybean industry.

[0003] Yield per plant is a core indicator for measuring soybean production potential, and its components include three major factors: number of pods per plant, number of seeds per pod, and seed weight. Among these, seed size / seed weight is a quantitative trait influenced by multiple factors. Existing research has revealed various regulatory factors affecting seed size, which collaboratively determine seed volume and weight by regulating the development of the seed coat, embryo, and endosperm. Researchers have located and cloned several major gene loci regulating seed size and weight using reverse or forward genetics methods. However, the number of key genes cloning seed weight regulation in soybean remains relatively limited, and its complex molecular regulatory network has not yet been fully elucidated, necessitating further in-depth mechanistic research.

[0004] Therefore, identifying key genes that synergistically regulate soybean grain weight and yield and elucidating their molecular mechanisms can have practical significance for improving soybean yield, and also provide important genetic resources and theoretical guidance for breeding high-yield soybean varieties. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides soybean GmMADS33 and its homologous genes and their applications, offering technical support for soybean seed and seedling cultivation and promoting the development of bio-agriculture and related industries.

[0006] The first objective of this invention is to provide soybean GmMADS33 protein, the amino acid sequence of which is shown in SEQ ID NO.3.

[0007] A second object of the present invention is to provide a coding gene encoding the GmMADS33 protein described above, the nucleotide sequence of which is shown in SEQ ID NO.1.

[0008] A third objective of this invention is to provide the application of the aforementioned GmMADS33 protein or the aforementioned encoding gene or related biological materials in regulating soybean yield.

[0009] Preferably, the application of increasing the expression level and / or activity of the GmMADS33 protein, or overexpressing the above-mentioned encoding gene, in increasing soybean yield.

[0010] Preferably, the application of reducing the expression level and / or activity of the GmMADS33 protein, or knocking out the coding gene, in reducing soybean yield.

[0011] Preferably, the expression level and / or activity of the GmMADS33 protein and its homolog GmMADS34 protein are reduced, or the coding gene and its homolog are knocked out. GmMADS34 The application of the gene in reducing soybean yield, the amino acid sequence of the GmMADS34 protein is shown in SEQ ID NO.4, the gene... GmMADS34 The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0012] Preferably, the soybean yield is at least one of soybean pod size, number of pods, number of seeds, and seed weight.

[0013] A fourth objective of this invention is to provide a method for regulating soybean yield, comprising the following steps: increasing the expression level and / or activity of the GmMADS33 protein in soybeans, or overexpressing the coding gene in soybeans to increase soybean yield; or decreasing the expression level and / or activity of the GmMADS33 protein in soybeans, or knocking out the coding gene in soybeans to decrease soybean yield.

[0014] Preferably, reducing the expression level and / or activity of the GmMADS33 protein in soybean further includes reducing the expression level and / or activity of its homolog GmMADS34 protein, the amino acid sequence of which is shown in SEQ ID NO. 4; the knockout of the above-mentioned coding gene further includes knocking out its homologous gene. GmMADS34 Genes, the ones mentioned GmMADS34 The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0015] Preferably, the soybean yield is at least one of soybean pod size, number of pods, number of seeds, and seed weight.

[0016] Advantages of this invention:

[0017] This invention provides soybeans GmMADS33 Genes and their homologous genes GmMADS34 And applications, compared to the cultivated soybean variety Williams (W82). gmmad33 Single protrusion and gmmads33 / gmmads34 Soybeans exhibited reduced pod width and projected area, as well as a decrease in 100-kernel weight; GmMADS33 -OE soybeans exhibit increased pod area, higher 100-seed weight, and increased number of pods, seeds, and yield per plant. Therefore, GmMADS33 These genes play an important role in regulating pod size, pod number, seed number, and seed weight, and are key regulatory genes for soybean pod size, seed weight, and seed number. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the GmMADS33 / GmMADS34 gene editing model, where red text indicates the knockout target sequence and mutated bases are represented by -.

[0019] Figure 2 for gmmads33 / gmmads34 Phenotypic diagram of transgenic knockout lines.

[0020] Figure 3 for GmMADS33 Phenotypic diagram of transgenic overexpression lines.

[0021] Figure 4 for GmMADS33 Image showing the identification results of transgenic overexpression lines. Detailed Implementation

[0022] The technical approach of this invention is as follows:

[0023] (1) Cloning from soybeans GmMADS33 Genes will GmMADS33 The genome sequence was recombined into a soybean transgenic vector to obtain an enhanced expression transformation vector. GmMADS33 -OE is used for soybean genetic transformation.

[0024] (2) Constructing a knockout GmMADS33 (The CDS sequence is shown in SEQ ID NO.1) and GmMADS34 A dual knockout vector (CDS sequence shown in SEQ ID NO.2) was used for soybean genetic transformation.

[0025] (3) The two transgenic vectors mentioned above were transferred into Agrobacterium, and the Agrobacterium carrying the vector plasmid was used for soybean genetic transformation.

[0026] (4) Transplanted into W82 cultivar soybeans, soybean cells regenerated into plants, and herbicide-resistant homozygous mutant plants were screened starting from the T0 generation to obtain GmMADS33 Transgenic soybean plants with enhanced and knockout gene expression.

[0027] (5) In T2 generation homozygous mutant plants, the soybean pod area was smaller and the 100-seed weight was lower, while in overexpression plants, the soybean pod area was increased, the 100-seed weight was increased, and the number of seeds was increased. Therefore, GmMADS33 These genes are key regulators of soybean pod size, seed weight, and seed number.

[0028] In some embodiments of the present invention, soybeans are disclosed. GmMADS33 Genes and GmMADS34 The application of genes in regulating soybean pod development, seed size and seed number, or in breeding high-yielding and high-quality soybean varieties, in the soybean GmMADS33The CDS sequence of the gene is shown in SEQ ID NO.1, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO.3; GmMADS34 The CDS sequence of the gene is shown in SEQ ID NO.2, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO.4.

[0029] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0030] Example 1: Cloning of GmMADS33 and GmMADS34 genes

[0031] This embodiment cloned the... GmMADS33 Genes and GmMADS34 Genes, including the following steps:

[0032] 1. Total RNA was extracted from soybean variety W82 seeds using the RNAprep Pure Plant Kit (Tiangen Biotech (Beijing) Co., Ltd., DP432). cDNA was obtained by reverse transcription using the PrimeScript™ II 1st strand cDNA Synthesis (TaKaRa) kit as a cloning template.

[0033] 2. Using cDNA as a template, PCR amplification was performed using primer pairs GmMADS33-F (SEQ ID NO. 5) and GmMADS33-R (SEQ ID NO. 6), and GmMADS34-F (SEQ ID NO. 7) and GmMADS34-R (SEQ ID NO. 8), respectively. The reaction system is shown in Table 1. The PCR amplification program was as follows: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 30 sec; 56℃ annealing for 30 sec; 72℃ extension for 1 min; 34 cycles; 72℃ extension for 5 min.

[0034] GmMADS33-F (SEQ ID NO.5):TCTCTAGGGTTCCAAGAGTGTCTTG

[0035] GmMADS33-R (SEQ ID NO.6): ATAACTAAAGAACAAGCAGCATT

[0036] GmMADS34-F (SEQ ID NO.7): TAAGAGAAGGAAAGAGGGGGATAGA

[0037] GmMADS34-R (SEQ ID NO.8):TCAAAGCATCCATCCAGGAA

[0038] 3. The PCR products were sent to Novizan Sequencing for sequencing.

[0039] Sequencing results show: GmMADS33 The CDS length is 750 bp (SEQ ID NO.1) and it can encode 249 amino acids (SEQ ID NO.3); GmMADS34 The CDS length is 750 bp (SEQ ID NO.2) and it encodes 249 amino acids (SEQ ID NO.4).

[0040] Example 2: Construction of GmMADS33 gene overexpression vector

[0041] 1. PCR amplification of the target gene fragment

[0042] The sequencing in Example 1 was used to verify the correctness. GmMADS33 Using the CDS clone plasmid of the gene as a template, PCR amplification was performed using the forward adapter primer GmMADS-PTF101-F (SEQ ID NO. 9) and the reverse adapter primer GmMADS-PTF101-R (SEQ ID NO. 10). The reaction system is shown in Table 2. The amplification program was as follows: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 30 sec; 55℃ annealing for 30 sec; 72℃ extension for 20 s, 34 cycles; 72℃ extension for 5 min. After amplification, the PCR products were detected by 1% agarose gel electrophoresis to confirm the product size and consistency with the target product. GmMADS33 The full-length (750 bp) gene CDS is identical.

[0043] GmMADS-PTF101-F (SEQ ID NO.9):

[0044] GAGAACACGGGGGACTCTAGA ATGGGGAGGGGAAGAGTGGAA

[0045] GmMADS-PTF101-R (SEQ ID NO.10):

[0046] GTCATCCTTGTAGTCCATGGATCC AAGCATCCATCCAGGAATAA

[0047] 2. Linearization of the carrier

[0048] In a 50 µL system, 1 µg of PTF101 vector and 1 µL each of Xba I and BamH I enzymes were added for double digestion. The reaction system is shown in Table 3. The digestion reaction was carried out at 37℃ for 1 h.

[0049] 3. The PCR product obtained in step 1 and the purified linearized vector obtained in step 2 were recovered and purified according to the kit instructions (Tiangen Biotech (Beijing) Co., Ltd., DP209-02).

[0050] 4. Homologous recombination reaction

[0051] The linearized vector recovered in step 3 was mixed with the PCR product of the target gene, and recombinase (Exnase II) was added. The reaction system is shown in Table 4. The reaction was carried out at 37℃ for 20 min to complete the recombination reaction and obtain the recombinant ligation product.

[0052] 5. Transformation of competent states and identification

[0053] The ligation product was transformed into *E. coli* TOP10 competent cells using a heat shock method and then plated for culture. The recombinant ligation product was mixed with TOP10 competent cells, incubated on ice for 30 min, heat-shocked at 42℃ for 90 s, and immediately incubated on ice for 2 min. Then, 500 μL of antibiotic-free LB broth was added, and the cells were incubated at 37℃ with shaking at 200 rpm for 1 h. After incubation, the cells were centrifuged at 5000 rpm for 1 min, 400 μL of supernatant was discarded, and the mixture was thoroughly mixed. 100 μL of the bacterial culture was plated onto LB solid medium and incubated upside down at 37℃. Positive clones were picked and expanded in LB broth containing 50 µg / mL SPE antibiotic. The bacterial culture was identified by PCR using primers 35S-P (SEQ ID NO. 11) and NOS-R (SEQ ID NO. 12), and then sequenced for verification. Bacterial cultures with correct sequencing were expanded and plasmids were extracted, thus obtaining the desired bacterial culture. GmMADS33 Overexpression vector.

[0054] 35S-P (SEQ ID NO.11): GACGCACAATCCCACTATCC

[0055] NOS-R (SEQ ID NO.12): CAAGACCGGCAACAGGATTCAA

[0056] Example 3: Construction of a dual knockout vector for the GmMADS33 / GmMADS34 gene

[0057] This embodiment has been constructed. GmMADS33 Genes and GmMADS34 A dual knockout vector for genes includes the following steps:

[0058] 1. In view of GmMADS33 and GmMADS34The nucleotide sequences are highly similar, and sequence 1 (GGATAGAGAACAAGATAAAC, SEQ ID NO.13) and sequence 2 (CGTTTGCAAAGAGGAGAAAT, SEQ ID NO.14) were selected as common knockout targets for both.

[0059] 2. Using the pGES401 vector as a template, PCR amplification was performed using the forward adapter primer GmMADS-pGES401-F (SEQ ID NO. 15) and the reverse adapter primer GmMADS-pGES401-R (SEQ ID NO. 16) to obtain the sgRNA expression cassette fragment carrying the target sequence. The reaction system is shown in Table 5. The amplification program was as follows: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 30 sec; 55℃ annealing for 30 sec; 72℃ extension for 20 s, 34 cycles; 72℃ extension for 5 min.

[0060] GmMADS-pGES401-F (SEQ ID NO. 15):

[0061] TGGTCTCGTGCA GGATAGAGAACAAGATAAAC GTTTTAGAGCTAGAAATAGC

[0062] GmMADS-pGES401-R (SEQ ID NO. 16):

[0063] TGGTCTCgAAAC ATTTCTCCTCTTTGCAAACG TGCACCAGCCGGGAATCGAA

[0064] 3. Purify the PCR products according to the instructions of the HiPure Gel Pure DNA Mini Kit (Magen, D2111-03).

[0065] 4. Golden Gate one-step carrier construction

[0066] The purified PCR products from step 3 (DNA fragment 1 and DNA fragment 2, i.e., sgRNA expression cassette fragments carrying two target sites) were mixed with the pGES401 vector plasmid, and T4 DNA ligase, BsaI-HFv2 restriction enzyme, and corresponding buffers were added to construct a Golden Gate one-step reaction system, as shown in Table 6. First, the reaction was incubated at 37℃ for 20 min; then, ligation was performed using variable-temperature cycling for 20 cycles: 37℃ for 2 min, 16℃ for 2 min; finally, the enzyme activity was inactivated at 37℃ for 15 min, followed by 85℃ for 5 min, terminating the reaction. The recombinant ligation product of the double knockout vector was obtained.

[0067] 5. Transformation of competent states and identification

[0068] The ligation product obtained in step 4 was transferred to *E. coli* TOP10 competent cells using a heat shock method and plated for culture: the recombinant ligation product was mixed with TOP10 competent cells, incubated on ice for 30 min, heat-shocked at 42℃ for 90 s, and immediately incubated on ice for 2 min; then 500 μL of LB liquid medium was added, and the cells were incubated at 37℃ and 200 rpm for 1 h with shaking; after incubation, the cells were centrifuged at 5000 rpm for 1 min, 400 μL of supernatant was discarded, the mixture was reconstituted by pipetting, and 100 μL of the bacterial culture was plated on LB solid medium. Positive clones were picked and expanded in LB medium containing 50 μg / mL Kan antibiotic working solution. PCR identification and sequencing verification were performed using primers STU-TEST-3F (SEQ ID NO.17) and STU-TEST-4R (SEQ ID NO.18), thereby obtaining... GmMADS33 and GmMADS34 Dual knockout vector.

[0069] STU-TEST-3F (SEQ ID NO.17): TGCTACCCTCATCCATCAGTC

[0070] STU-TEST-4R (SEQ ID NO.18): TGTTGTGTGGAATTGTGAGCG

[0071] Example 4: Agrobacterium-mediated transformation of soybean

[0072] The plasmids that were correctly sequenced and constructed in Examples 2 and 3 were sent to Wuhan Boyuan Biotechnology Co., Ltd. for the construction of transgenic lines.

[0073] Example 5: Identification and Phenotypic Analysis of Transgenic Lines

[0074] 1. Molecular identification

[0075] Overexpression lines: Total protein was extracted from the test lines and the empty vector control (Williams, W82), subjected to SDS-PAGE electrophoresis, and transferred to a membrane. The membrane was then incubated with a commercially available anti-Flag tag antibody as the primary antibody, followed by mouse secondary antibody, and finally analyzed by chemiluminescence. To ensure reliable results, the internal control protein (Actin) was simultaneously detected to correct for the loading amount. The criteria for a positive overexpression line were: the appearance of a specific band at the expected molecular weight position, with a signal intensity significantly stronger than that of the empty vector control.

[0076] Knockout lines: PCR amplification was performed using specific primers MADS33-JD-F1 (SEQ ID NO.19), MADS33-JD-R1 (SEQ ID NO.20), MADS34-JD-F1 (SEQ ID NO.21), and MADS34-JD-R1 (SEQ ID NO.22) designed for the editing site. The products were then sequenced to confirm the expected deletion, insertion, or substitution of the gene sequence.

[0077] MADS33-JD-F1 (SEQ ID NO.19): ATAAAAAAGCCAAGTTTGATAGACA

[0078] MADS33-JD-R1 (SEQ ID NO.20):AATATAAGCAAATCAAAGGAGAGCC

[0079] MADS34-JD-F1 (SEQ ID NO.21): TAAGAGAAGGAAAGAGGGGGATAGA

[0080] MADS34-JD-R1 (SEQ ID NO.22): ACACTCCAGCATACGAAAGAG

[0081] 2. Confirmation of genetic stability

[0082] The T0 generation plants that tested positive were self-pollinated to obtain T1 and T2 generation seeds. Molecular identification was then performed in subsequent generations to screen for lines that stably inherited the transgene and were homozygous for subsequent phenotypic analysis.

[0083] gmmads33 / gmmads34 The results of the identification of transgenic mutant lines are as follows: Figure 1 As shown, GmMADS33 The results of the identification of transgenic overexpression lines are as follows: Figure 4 As shown, it indicates gmmads33 / gmmads34 Transgenic mutant lines and GmMADS33 The transgenic overexpression line was successfully constructed. gmmads33 / gmmads34 Phenotypic characteristics of transgenic knockout lines, such as Figure 2 As shown, compared to the cultivated soybean variety Williams (W82). gmmad33 Single protrusion and gmmads33 / gmmads34 The soybeans exhibited reduced pod width and projected area, as well as a decrease in 100-kernel weight. GmMADS33 Phenotypic characteristics of transgenic overexpression lines, such as Figure 3 As shown, compared to the control cultivar Williams (W82). GmMADS33-OE soybeans showed increased pod area, higher 100-seed weight, and increased number of pods, seeds, and yield per plant. These results indicate that: GmMADS33 Genes play an important role in regulating pod size, pod number, seed number, and seed weight.

[0084] SEQ ID NO.1 ( GmMADS33 (CDS sequence of the gene) ATGGGGAGGGGAAGAGTGGAA CTCAAAA GGATAGAGAACAAGATAAAC AGGCAGGTAA CGTTTGCAAA GAGGAGAAAT GGGCTTCTCAAGAAAGCCTATGAGCTTTCTGTTCTCTGTGATGCTGAGGTTGCCCTCATCATCTTCTCTACCCGCGGCAAGCTTTATGAATTCTGTAGCACCAACAGCATGCTCAAAACACTTGAAAGGTACCAAAAGTGCAGCTATGGTGCAGTTGAAGTC AGCAAACCTGGCAAAGAGCTTGAGCAGAGCAGCTACCGTGAGTACTTGAAGCTGAAAGCAAGATTTGAATCTCTTCAAAGGACCCAAAGGAACCTTCTTGGTGAAGACTTGGGCCCACTAAATACCAAAGATCTTGAGCAGCTTGAGCGGCAACTGGATTCA TCTCTCAAGCAAGTGAGGTCCACAAAGACTCAGTTCATGCTGGACCAGTTAGCTGATCTTCAGAATAAGGAGCATATGCTGGTAGAAGCAAACAGATCTTTGACCATGAAGCTGGAAGAAATCAATTCAAGAAACCAGTATAGGCAAACATGGGAAGCTGGT GAGCAAAGTATGTCATATGGTACACAAAATGCTCACTCCCAAGGCTTTTTCCAGCCTTTGGAGTGCAACCCCACATTGCAGATAGGTTCTGACTACAGGTACAACCCTGAAGCCTCAGAGCAGCAGCTAGCTGCCACAACTCAAGCTCAACAAGTCAATGGAT TTATTCCTGGATGGATGCTT TGA.

[0085] SEQ ID NO.2 ( GmMADS34 (CDS sequence of the gene) ATGGGGAGGGGAAGAGTGGAACTGAAAAGGATAGAGAACAAGATAAAC AGGCAGGTAA CGTTTGCAAA GAGGAGAAAT GGGCTTCTCAAGAAAGCCTATGAGCTTTCTGTTCTCTGTGATGCTGAGGTTGCCCTCATCATCTTCTCCACCCGTGGCAAGCTTTATGAATTCTGCAGCACCAACAGCATGCTCAAAACACTTGAAAGGTACCAAAAGTGCAGCTATGGTGCAGTGGAAGTCAGCAAACCTGGCAAAGAGCTTGAGCAGAGCAGCTACCGTGAGTACTTGAAGCTGAAAGCAAGATTTGAATCTCTTCAAAGGACCCAAAGGAACCTTCTTGGTGAAGACTTGGGCCCATTAAATACCAAAGATCTTGAGCAGCTTGAGCGCCAACTGGATTCATCTCTTAAGCAAGTGAGGTCAACAAAGACTCAGTTCATGCTGGACCAGTTAGCTGATCTTCAGAATAAGGAGCATATGTTGGTGGAAGCAAACAGATCTTTGACCATGAAGCTGGAAGAAATCAATTCAAGAAACCAGTATAGGCAAACATGGGAAGCTGGTGAGCAAAGTATGCCATATGGAACCCAAAATGCTCACTCTCAAGGCTTCTTCCAGCCTTTGGAGTGCAACCCCACATTGCAGATAGGTTCTGACTACAGGTACATCCCTGAAGCCTCAGAGCAGCAGCTAGCTGCCACAACTCAAGCTCAACAAGTCAATGGATTTA TTCCTGGATGGATGCTTTGA 。

[0086] SEQ ID NO.3 (Amino acid sequence of GmMADS33) MGRGRVELKRIENKINRQVTFAKRRNGLLKKAYELSVLCDAEVALIIFSTRGKLYEFCSTNSMLKTLERYQKCSYGAVEVSKPGKELEQSSYREYLKLKARFESLQRTQRNLLGEDLGPLNTKDL EQLERQLDSSLKQVRSTKTQFMLDQLADLQNKEHMLVEANRSLTMKLEEINSRNQYRQTWEAGEQSMSYGTQNAHSQGFFQPLECNPTLQIGSDYRYNPEASEQQLAATTQAQQVNGFIPGWML.

[0087] SEQ ID NO.4 (amino acid sequence of GmMADS34) MGRGRVELKRIENKINRQVTFAKRRNGLLKKAYELSVLCDAEVALIIFSTRGKLYEFCSTNSMLKTLERYQKCSYGAVEVSKPGKELEQSSYREYLKLKARFESLQRTQRNLLGEDLGPLNTKDL EQLERQLDSSLKQVRSTKTQFMLDQLADLQNKEHMLVEANRSLTMKLEEINSRNQYRQTWEAGEQSMPYGTQNAHSQGFFQPLECNPTLQIGSDYRYIPEASEQQLAATTQAQQVNGFIPGWML.

[0088] Table 1

[0089] The same reaction system was used for PCR amplification of GmMADS33 and GmMADS34.

[0090] Table 2

[0091] Table 3

[0092] Table 4

[0093] Table 5

[0094] Table 6

Claims

1. Soybean GmMADS33 protein, the amino acid sequence of which is shown in SEQ ID NO.

3.

2. The gene encoding the GmMADS33 protein of claim 1, characterized in that, The nucleotide sequence is shown in SEQ ID NO.

1.

3. The application of the GmMADS33 protein of claim 1 or the encoding gene of claim 2 or related biological materials in regulating soybean yield.

4. The application according to claim 3, characterized in that, This refers to the application of increasing the expression level and / or activity of the GmMADS33 protein as described in claim 1, or overexpressing the encoding gene as described in claim 2, in increasing soybean yield.

5. The application according to claim 3, characterized in that, This refers to the application of reducing the expression level and / or activity of the GmMADS33 protein as described in claim 1, or knocking out the encoding gene as described in claim 2, in reducing soybean yield.

6. The application according to claim 5, characterized in that, This involves reducing the expression level and / or activity of the GmMADS33 protein and its homolog GmMADS34 protein as described in claim 1, or knocking out the encoding gene and its homolog as described in claim 2. GmMADS34 The application of the gene in reducing soybean yield, the amino acid sequence of the GmMADS34 protein is shown in SEQ ID NO. 4, the gene... GmMADS34 The nucleotide sequence of the gene is shown in SEQ ID NO.

2.

7. The application according to any one of claims 3-6, characterized in that, The soybean yield is defined as at least one of the following: soybean pod size, number of pods, number of seeds, and seed weight.

8. A method for regulating soybean yield, characterized in that, The method includes the following steps: increasing the expression level and / or activity of the GmMADS33 protein of claim 1 in soybeans, or overexpressing the coding gene of claim 2 in soybeans to increase soybean yield; or decreasing the expression level and / or activity of the GmMADS33 protein of claim 1 in soybeans, or knocking out the coding gene of claim 2 in soybeans to decrease soybean yield.

9. The method according to claim 8, characterized in that, The reduction of the expression level and / or activity of the GmMADS33 protein of claim 1 in soybeans further includes reducing the expression level and / or activity of its homolog GmMADS34 protein, the amino acid sequence of which is shown in SEQ ID NO.4; The knockout of the coding gene of claim 2 further includes knocking out its homologous genes. GmMADS34 Genes, the ones mentioned GmMADS34 The nucleotide sequence of the gene is shown in SEQ ID NO.

2.

10. The method according to claim 8 or 9, characterized in that, The soybean yield is defined as at least one of the following: soybean pod size, number of pods, number of seeds, and seed weight.

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