Application of GmATL3 in regulation and control of soybean grain size

Abstract

The invention discloses application of GmATL3 in regulation and control of soybean grain size, and belongs to the technical field of gene engineering, the amino acid sequence of a protein encoded by the GmATL3 gene is shown as SEQ ID NO.1, the CDS sequence of the GmATL3 gene is shown as SEQ ID NO.2, and the full-length nucleotide sequence of the GmATL3 gene is shown as SEQ ID NO.3. The invention further discloses a preparation method of the GmATL3 gene. Experiments prove that the gene or the encoded protein thereof has a positive regulation effect on the soybean grain size, so that a new thought is provided for cultivating high-yield soybeans.

Description

Technical Field

[0001] This application belongs to the field of genetic engineering technology, specifically involving the application of GmATL3 in regulating soybean seed size. Background Technology

[0002] The main components of soybean (Glycine max) yield include plant number per unit area, number of seeds per plant, and 100-seed weight. Among these, seed size is not only one of the key agronomic traits in domestication and selection, but also the most direct indicator of 100-seed weight. Therefore, increasing soybean seed size is a crucial way to improve soybean yield, and identifying functional genes related to soybean seed size and their molecular regulatory mechanisms is of great significance for increasing soybean yield.

[0003] Previous studies have shown that zinc finger proteins (ZFPs) play an important role in plant development. Therefore, studying the RING-ZFP family members in soybean and identifying new functional genes significantly related to soybean seed size is of great significance for high-yield soybean breeding. Summary of the Invention

[0004] In view of this, the primary objective of this application is to provide the application of GmATL3 in regulating soybean seed size. This application has discovered a functional gene, GmATL3, that positively regulates soybean seed size. By constructing materials with gene knockout and overexpression, its regulatory role in soybean seed size has been determined, which is of great significance for breeding high-yield soybeans.

[0005] To achieve the above objectives, this application adopts the following technical solution: One aspect of this application discloses the application of the GmATL3 gene or its encoded protein in regulating soybean seed size, wherein the amino acid sequence of the protein encoded by the GmATL3 gene is shown in SEQ ID NO.1.

[0006] Another aspect of this application discloses a recombinant vector containing nucleotide molecules that enhance the expression of the GmATL3 gene.

[0007] Another aspect of this application discloses a transformant obtained by introducing the recombinant vector described in this application into a host cell and then transforming it.

[0008] Another aspect of this application discloses the use of upregulating molecules, recombinant vectors, or transformants that increase the expression of the GmATL3 gene or its encoded protein as described in this application in increasing soybean seed size, preparing products with increased soybean seed size, or preparing high-yield soybeans.

[0009] Another aspect of this application discloses a kit containing the recombinant vector or transformant described in this application.

[0010] Another aspect of this application discloses a method for breeding high-yield soybeans or increasing soybean yield, comprising the step of increasing the expression of the GmATL3 gene or the protein encoded therein as described in this application in soybean plants.

[0011] The beneficial effects of this application are: The GmATL3 gene or its encoded protein disclosed in this application has a positive regulatory effect on soybean seed size and yield. Overexpression of the GmATL3 gene or its encoded protein results in larger soybean seeds. Specifically, compared to wild-type soybeans, soybeans overexpressing the GmATL3 gene or its encoded protein exhibit significantly increased seed width, length, and seed weight (e.g., 100-seed weight / 1000-seed weight). This novel discovery of the function of the GmATL3 gene or its encoded protein provides a new method for increasing soybean yield and cultivating high-yield soybeans. Attached Figure Description

[0012] Figure 1 This is a knockout form of the GmATL3 gene.

[0013] Figure 2 For WT and atl3 Analysis of the morphology and phenotypic characteristics of mature seeds of mutants, including... Figure 2 'a' is atl3 The mature grain morphology of the three knockout plants; Figure 2 In the middle b, the statistical analysis results of the grain size trait are shown; among them, atl3 The knockout mutant material showed significantly reduced weight per 100 grains, grain length, and grain width. express p<0.01 .

[0014] Figure 3 This is a map of the pCAMBIA1305-EGFP vector.

[0015] Figure 4 Phenotypic analysis of Arabidopsis plants overexpressing GmATL3; among which, Figure 4 In the middle, a represents the increased plant height of Arabidopsis thaliana overexpressing GmATL3; Figure 4 In the middle b, the root length of Arabidopsis thaliana overexpressing GmATL3 showed a highly significant difference on day 6. Figure 4 In the middle (c), the overexpressing Arabidopsis seeds were significantly larger; express p<0.01 .

[0016] Figure 5 For quantitative trait analysis of Arabidopsis thaliana overexpressing GmATL3; among which, Figure 5In Figure 'a', the leaves of Arabidopsis thaliana plants overexpressing GmATL3 are enlarged; Figure 5 In the middle b, the thousand-seed weight, seed length, and seed width of Arabidopsis thaliana overexpressing GmATL3 were significantly increased. express p＜0.01 . Detailed Implementation

[0017] The embodiments of this application will be clearly and completely described below. The technical solutions in the embodiments described below are exemplary and only possible technical implementations of this application, not all possible implementations. Those skilled in the art can combine the embodiments of this application to obtain other embodiments without creative effort, and these embodiments are also within the protection scope of this application.

[0018] Through preliminary research and analysis, this application identified the functional gene GmATL3, which is significantly associated with soybean seed size. GmATL3 or its encoded protein has a positive regulatory effect on soybean seed size. This regulation refers to overexpression of GmATL3 or its encoded protein, i.e., increasing the expression level or activity of the GmATL3 gene or its encoded protein, resulting in an increase in soybean seed weight (100-seed weight or 1000-seed weight), as well as a significant increase in seed width and length.

[0019] In this application, for the sake of simplicity, the names of proteins and genes are used interchangeably. For example, GmATL3 can refer to a gene or the protein encoded by that gene. Those skilled in the art will understand that they represent different substances in different descriptive contexts. For instance, in this application, when GmATL3 is described as a protein, it refers to the protein with the amino acid sequence SEQ ID NO.1. When GmATL3 is described as a gene, it refers to the gene with the CDS sequence SEQ ID NO.2 or the full-length nucleotide sequence SEQ ID NO.3. In this application, the soybean reference genome version is Wm82.a6.v1. This is readily understood by those skilled in the art.

[0020] This application further discloses a recombinant vector and a transformant, wherein the recombinant vector contains nucleotide molecules that enhance the expression of the GmATL3 gene. The transformant is obtained by introducing the aforementioned recombinant vector into host cells and then transforming them.

[0021] The "recombinant vector" described in this application is formed by effectively ligating the nucleotide sequence molecule of the GmATL3 gene into an expression vector. The expression vector can be any of a virus, plasmid, bacteriophage, etc.; preferably, the expression vector is a plasmid, and there is no particular limitation on the specific type. Any general-purpose expression vector in the art or a self-developed vector can be used, as long as the GmATL3 gene can be inserted into it and it can perform its correct biological function. In some examples of this application, the expression vector is the standard general-purpose vector pCAMBIA1305-EGFP.

[0022] The "transformer" described in this application refers to the recombinant vector introduced into a host cell using genetic engineering techniques known in the art or developed independently, which enables the GmATL3 gene to be stably expressed in the host cell.

[0023] In this application, the term "host cell" is not particularly limited, but is preferably a bacterial cell, such as *Escherichia coli* or *Agrobacterium*. Specific transformation methods can be those known in the art or those developed in-house. For example, in some specific examples, *Agrobacterium*-mediated transgenic technology can be used to introduce the recombinant vector into soybean plants.

[0024] This application further discloses the use of upregulating molecules that increase the expression of the GmATL3 gene or its encoded protein, or recombinant vectors, or transformants in increasing soybean seed size, or in preparing products with increased soybean seed size, or in preparing high-yield soybeans.

[0025] In this application, the term "upregulation molecule" refers to a reagent that can specifically increase the expression of the GmATL3 gene or its encoded protein. Specific examples include, but are not limited to, protein or DNA activators, small molecule compounds, etc., which can be selected or designed by those skilled in the art based on actual needs.

[0026] This application further discloses a method for cultivating high-yield soybeans or increasing soybean yield, comprising the step of increasing the expression of the GmATL3 gene or its encoded protein in soybean plants.

[0027] In particular, increasing the expression of the GmATL3 gene or its encoded protein can be achieved using genetic engineering techniques well-known in the field or independently developed, without any special limitations. By increasing the expression of the GmATL3 gene or its encoded protein, soybean yield can be increased.

[0028] In some specific examples, the method mainly includes the following steps: Constructing biomaterials; The biological material is introduced into the starting crop to obtain soybeans with increased yield compared to the starting crop. The soybeans with increased yield are characterized by increased grain weight (e.g., 100-grain weight / 1000-grain weight), increased grain width, and increased grain length.

[0029] The “biomaterials” mentioned in this application refer to the aforementioned recombinant vectors or transformants introduced into the starting crop, such as Agrobacterium-mediated transformation technology commonly used in the art, thereby obtaining soybeans with increased yield.

[0030] This application further discloses a kit containing the recombinant vector or transformant described in this application. This kit can be used to cultivate high-yield soybeans or improve soybean yield.

[0031] It is understood that, in addition to the recombinant vector or transformant described herein, the kit may also contain other auxiliary reagents as needed. Furthermore, it may include an instruction manual, which should specify how to use the kit.

[0032] The present application will be further illustrated below with reference to specific embodiments. It should be noted that the specific embodiments below are for illustrative purposes only and do not limit the scope of the present application in any way.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for the purpose of describing particular embodiments only and is not intended to be limiting of this application.

[0034] In addition, unless otherwise specified, methods without detailed conditions or steps are conventional methods, and the reagents and materials used are commercially available.

[0035] Example 1: Discovery of GmATL3 candidate genes In previous studies, 146 soybean germplasm accessions from 14 environments across three major ecological regions in China were collected. Genome-wide association study (GWAS) analysis was used to identify the SW9-1 locus in these germplasm samples. Furthermore, a population of recombinant inbred lines (RILs) was constructed for fine mapping of candidate genes. Combined with transcriptome sequencing and weighted gene co-expression network analysis (WGCNA), the key gene GmATL3 regulating grain size was screened out.

[0036] Example 2 GmATL3 knockout material— atl3 mutant The tested soybean variety was Williams 82. Using CRISPR / Cas9 gene editing technology, four target sites were designed to knock out GmATL3 in Williams 82 as the background, resulting in GmATL3 knockout. atl3 Mutants. The soybeans used in the test were provided by the Soybean Molecular Breeding Research Group of Anhui Agricultural University. The knockout target sites are shown in Table 1: Table 1 Knockout target sequences

[0037] Extracted by CTAB method atl3 Using mutant leaf DNA as a template, the GmATL3 gene sequence was cloned using the specific primers in Table 2.

[0038] Table 2 Primer Information

[0039] PCR amplification was performed using the high-fidelity enzyme Primer STAR Max Premix (2×) from TAKARA, following the reaction system in Table 3 and the reaction procedure in Table 4: Table 3 PCR reaction system

[0040] Table 4 PCR reaction procedure

[0041] The obtained PCR products were detected by agarose gel electrophoresis at a mass percentage of 1.5%.

[0042] The PCR products were sent to a third-party commissioned agency, Shanghai Bioengineering, for sequencing, and the gene editing method was identified by sequence alignment using SnapGene.

[0043] pass Figure 1 It can be seen that, atl3 The GmATL3 gene in the mutant lacks the CGTCTTCACC fragment at 318bp, indicating the successful construction of the knockout material.

[0044] Example 3 atl3 Mutant phenotypic identification Cultivating soybean plants (including wild-type soybeans and) using nutrient soil (black soil: vermiculite in a 3:1 ratio) atl3 The mutant material was grown in the soybean greenhouse of Anhui Agricultural University (20,000 Lux, 70% humidity, 28℃ / 8h during the day and 26℃ / 16h at night).

[0045] Record the morphology of soybean plants at different stages; harvest the seeds after the plants mature and place them in a 40℃ oven for 3 hours to dehydrate; determine the weight of 100 seeds using a 0.01% balance.

[0046] Particle length and width were determined using ImageJ software; statistical analysis was performed using SPSS software; and tissue sections were analyzed using CaseViewer.

[0047] The results are as follows Figure 2 As shown, WT and atl3 The 100-grain weights were 17.94±0.13g and 14.71±0.59g, respectively; WT and atl3 The grain lengths were 0.81±0.03cm and 0.76±0.02cm, respectively; WT and atl3 The particle widths were 0.76±0.02cm and 0.63±0.03cm, respectively.

[0048] Compared to WT, atl3 The weight of 100 grains decreased by 18.00%, which was highly significant. p＜0.01 The grain length and grain width decreased by 6.14% and 16.36% respectively, with highly significant differences. p＜0.01 ).

[0049] Example 4 GmATL3-OE Construction of Arabidopsis thaliana 4.1 Experimental Materials Wild-type Arabidopsis thaliana Col-0; expression vector pCAMBIA1305-EGFP; competent Escherichia coli cells Mach1-T1; Agrobacterium tumefaciens GV3101.

[0050] 4.2 Construction GmATL3-OE Arabidopsis (1) Amplification of the GmATL3 fragment Leaf DNA was extracted from wild-type (WT) soybeans and used as a template to amplify the GmATL3 fragment using the method described in Example 2.

[0051] (2) Vector primer design: In the pCAMBIA1305-EGFP vector (see diagram) Figure 3 Based on the Spe I and BamHI restriction sites in the d35S-EGFP region, a GmATL3 fragment was designed for insertion. The complete sequence (starting and ending 20 bp upstream and downstream of the restriction sites) and the target gene coding region sequence were uploaded to the Vazyme website to obtain the vector primers. The specific primer sequences are shown in Table 5. Table 5 Primer Sequences

[0052] (3) Vector double enzyme digestion The pCAMBIA1305-EGFP vector was double-digested according to the system in Table 6 (37℃, 4h; 80℃, 20min) to obtain the double-digested product.

[0053] Table 6 Double enzyme digestion system

[0054] (4) Homologous recombination Homologous recombination was performed on the DNA purified product from step (1) and the double enzyme digestion product from step (3) using the ClonExpress II One Step Cloning Kit (Vazyme). The reaction mixtures were prepared on ice, centrifuged, and then incubated at 37°C for 30 min in a PCR instrument. The resulting ligation products were immediately cooled on ice or stored in a -20°C freezer. The recombination systems are shown in Table 7. Table 7 Homologous Recombination System

[0055] (5) Escherichia coli transformation Mach-T1 competent cells were stored at -80°C and thawed on ice. 1 μL of the ligation product from step (4) was added and the mixture was gently stirred at the bottom of the centrifuge tube. After 25 min on ice, the mixture was transferred to a water bath and heat-shocked at 42°C for 45 s. It was then quickly returned to ice and allowed to stand for 2 min. In a pre-sterilized laminar flow hood, 700 μL of antibiotic-free LB liquid medium was added to the mixture, mixed, and placed in a 37°C shaker at 200 r / min for 1 h to recover. The centrifuge tubes were centrifuged at 5000 r / min for 1 min to collect the bacteria. Approximately 100 μL of supernatant was collected and gently mixed by pipetting. The mixture was then plated on LB solid medium containing Kansas antibiotics. The plates were incubated upside down in a 37°C incubator for approximately 12 hours. Single colonies (round) were picked from a clean bench and placed in a 50 mL centrifuge tube containing 20 mL of LB broth containing Kans antibiotic. The tube was then incubated upside down in a 37°C shaker at 200 rpm for 9 hours. Positive colonies were detected using PCR and sequenced. The correct bacterial culture was then preserved (50% glycerol and bacterial culture were mixed 1:1 and stored at -80°C in a clean bench).

[0056] (6) Extraction of Escherichia coli plasmids The experiment used the AxyPrep Plasmid Miniprep Kit for plasmid extraction. 4 mL of the above-mentioned *E. coli* bacterial culture was added in fractions to 2 mL sterile, enzyme-free centrifuge tubes. The tubes were centrifuged at 12000 rpm for 1 min, and the supernatant was discarded. 250 μL of Buffer S1 was added to each centrifuge tube and mixed by pipetting. Then, 250 μL of Buffer S2 was added, and the tubes were inverted 4-6 times to fully lyse the bacteria until a clear solution was formed. 250 μL of Buffer S3 was added to each centrifuge tube, and the tubes were inverted 4-6 times to mix (until white flocculent material appeared). The mixture was then transferred to a centrifuge and centrifuged at 12000 rpm for 10 min.

[0057] (7) Agrobacterium competent transformation Agrobacterium competent cells (GV3101) were stored at -80°C. After thawing on ice, 0.4 μL of recombinant plasmid was added and gently mixed. The mixture was then placed on ice for 5 min, frozen in liquid nitrogen for 5 min, placed in a 37°C water bath for 5 min, and placed on ice for 5 min. In a clean bench, 700 μL of antibiotic-free YEP medium was added to the mixture, and the mixture was incubated at 220 rpm for 2 h at 28°C. The cells were collected by centrifugation at 5000 rpm for 1 min, and transferred to a clean bench. Approximately 100 μL of supernatant was collected and gently mixed by pipetting. The mixture was then plated onto YEP solid medium containing Kan and Rif antibiotics. The medium was inverted and incubated at 220 rpm for 20-24 h at 28°C. Positive colonies were detected and sequenced using bacterial culture PCR. The correct bacterial culture was preserved (50% glycerol and bacterial culture were mixed 1:1 and stored at -80℃ in a clean bench). The supernatant was transferred to a 2mL preparation tube and centrifuged at 12000 rpm for 1 min, discarding the filtrate. The preparation tube was returned to the centrifuge tube, 500 μL of Buffer W1 was added, and the mixture was centrifuged at 12000 rpm for 1 min, discarding the filtrate. The preparation tube was returned to the centrifuge tube, 700 μL of Buffer W2 was added, and the mixture was centrifuged at 12000 rpm for 1 min, discarding the filtrate; this process was repeated once. The preparation tube was returned to a 2mL centrifuge tube and centrifuged at 12000 rpm for 2 min. The preparation tube was transferred to a new 1.5mL centrifuge tube, 60 μL of ddH2O was added to the center of the membrane, and the mixture was incubated at room temperature for 2 min; then centrifuged at 12000 rpm for 1 min, discarding the preparation tube to obtain the target plasmid DNA. After measuring the concentration and mass using a micro spectrophotometer, store in a -20°C refrigerator.

[0058] (8) Arabidopsis thaliana cultivation Wild-type Arabidopsis thaliana (Col-0) was cultivated using the soil culture method. Sterilized vermiculite and nutrient soil were mixed in a 3:1 ratio to form a substrate, which was then placed in small flower pots. Arabidopsis thaliana seeds were evenly scattered on the soil surface, covered with a film, and then punched with holes. The pots were placed in a 22℃ light incubator (16h light / 8h darkness, light intensity: 30000 Lux, humidity: 60%) and transplanted after 7 days of growth.

[0059] (9) Flower dipping method Transfer 1 mL of the *Agrobacterium* bacterial culture containing the constructed plant overexpression vector to a 15 mL centrifuge tube containing 2 mL of YEP medium (Kan, Rif) and incubate at 28°C for 24 h in a shaker at 220 rpm. After turbidity, transfer the culture to a 50 mL centrifuge tube containing 10 mL of YEP medium (Kan, Rif) and incubate at 28°C for 24 h in a shaker at 220 rpm. Centrifuge the cultured bacterial culture at 5000 rpm for 10 min, collect the bacteria and discard the supernatant. Wrap the centrifuge tube in aluminum foil to protect it from light. Add the prepared inoculation solution to adjust the OD to approximately 1.3 (600°C). Select healthy, appropriately grown wild-type *Arabidopsis* plants, remove any pods and flowering inflorescences, and use these for infection. Immerse the unflowered inflorescences in the prepared bacterial solution for 1 min and then treat them in a dark environment for one day. Repeat the inoculation process three times, with a one-week interval between each inoculation. Water the plants the day before each inoculation.

[0060] (10) Identification of homozygous overexpression Arabidopsis thaliana Infected Arabidopsis seeds (T1) were collected, and positive Arabidopsis were selected using MS medium with hygromycin resistance. Plants with good growth after 7 days were identified as positive plants and transplanted into prepared nutrient soil for cultivation. Seeds were harvested from each individual plant to screen for positive plants, and homozygous overexpression lines were obtained by the T3 generation.

[0061] (11) Arabidopsis seed germination Select dried Arabidopsis thaliana seeds and sterilize them in a clean bench according to the following steps: Place approximately 100 Arabidopsis thaliana seeds into a 1.5 mL centrifuge tube; add 12% sodium hypochlorite solution to the centrifuge tube, place in a shaker to wash for 15 min, centrifuge for 1 s; discard the supernatant, add 1000 μL of sterilized ddH2O, place in a shaker to wash for 3 min, centrifuge for 30 s, repeat this step 7 times. Using a sterilized pipette tip, aspirate Arabidopsis thaliana seeds and streak them individually on MS medium; after completion, seal the plate with aluminum foil to protect it from light, invert it in a 4℃ refrigerator for vernalization for 3 days, then transfer it to a plant tissue culture incubator. After 7 days of growth, transplant it into nutrient soil.

[0062] Example 5 GmATL3-OE Arabidopsis thaliana phenotypic identification The one constructed in Example 4 GmATL3-OEPhenotypic identification was performed on Arabidopsis thaliana and wild-type Arabidopsis thaliana, and the results are as follows: Figure 4 and Figure 5 As shown.

[0063] Phenotypic identification revealed that 6-day WT and GmATL3 The root lengths of -OE were 2.80±0.21cm and 3.24±0.14cm, respectively; compared to WT, GmATL3 -OE root length increased by 15.71%, and the difference was highly significant. p＜0.01 ).

[0064] Measurement of mature grain size phenotype revealed that WT and GmATL3 The thousand-seed weights of -OE plants were 23.13±0.95 mg and 25.99±1.24 mg, respectively; compared to WT, GmATL3 -OE increased the thousand-grain weight by 12.36%, which was highly significant. p ＜0.01 ).

[0065] WT and GmATL3 The kernel lengths of -OE seeds were 483.43±35.57μm and 599.65±47.19μm, respectively; compared to WT, GmATL3 -OE increased particle length by 24.04%, which was highly significant. p＜0.01 ).

[0066] WT and GmATL3 The kernel widths of -OE kernels were 290.61±19.96μm and 347.97±30.90μm, respectively; compared to WT, GmATL3 -OE increased the particle width by 19.74%, which was highly significant. p＜0.01 ).

[0067] The above examples demonstrate that GmATL3 is a positive regulator of soybean seed size, and overexpression of GmATL3 can increase soybean yield.

[0068] It should be noted that this application is not limited to the above-described embodiments. The above embodiments are merely examples, and any embodiments with the same structure and effect as the technical concept within the scope of this application are included in the technical scope of this application. Furthermore, various modifications that can be conceived by those skilled in the art to the embodiments, and other ways of constructing by combining some of the constituent elements of the embodiments, without departing from the spirit of this application, are also included in the scope of this application.

Claims

1. The application of the GmATL3 gene or its encoded protein in regulating soybean seed size, characterized in that, The amino acid sequence of the protein encoded by the GmATL3 gene is shown in SEQ ID NO.

1.

2. The application as described in claim 1, characterized in that, The CDS sequence of the GmATL3 gene is shown in SEQ ID NO.2; its full-length nucleotide sequence is shown in SEQ ID NO.

3.

3. The application as described in claim 1, characterized in that, The regulation refers to increasing the expression level of GmATL3 or its encoded protein, thereby increasing the weight of soybean seeds and / or the width and length of soybean seeds.

4. A recombinant vector, characterized in that, The recombinant vector contains nucleotide molecules that enhance the expression of the GmATL3 gene as defined in claim 1 or 2.

5. A transformant, characterized in that, The transformant is obtained by introducing the recombinant vector of claim 4 into a host cell and then transforming it.

6. An upregulating molecule that enhances the expression of the GmATL3 gene or its encoded protein as defined in claim 1 or 2, or the recombinant vector of claim 4, or the transformant of claim 5, in increasing soybean seed size, or in preparing products with increased soybean seed size, or in preparing high-yield soybeans.

7. A reagent kit, characterized in that, The kit contains the recombinant vector of claim 4 or the transformant of claim 5.

8. A method for cultivating high-yield soybeans or increasing soybean yield, characterized in that, include: The step of increasing the expression of the GmATL3 gene or the protein encoded by it as defined in claim 1 or 2 in soybean plants.

9. The method as described in claim 8, characterized in that, To achieve increased expression of the GmATL3 gene or its encoded protein, the following steps are included: Constructing biomaterials, wherein the biomaterials are the recombinant vector of claim 4 or the transformant of claim 5; The biological material was introduced into the starting crop to obtain soybeans with increased yield compared to the starting crop.

10. The method as described in claim 9, characterized in that, The increased yield of soybeans is manifested by an increase in soybean seed weight, and / or an increase in seed width and length.

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