Soybean GmLEC1 gene and application of encoded protein thereof

Abstract

The invention discloses a soybean GmLEC1 gene and application of an encoded protein thereof, the CDS sequence of the soybean GmLEC1 gene is shown as SEQ ID NO: 1 or SEQ ID NO: 2, and the amino acid sequence of the encoded protein of the soybean GmLEC1 gene is shown as SEQ ID NO: 3 or SEQ ID NO: 4. The invention discloses the biological functions of the soybean GmLEC1 gene in regulation and control of soybean grain size, hundred-grain weight, grease content and protein content for the first time, valuable gene resources and theoretical basis are provided for crop breeding, and the soybean GmLEC1 gene can be widely applied to high-yield and high-quality breeding of soybeans. And the seeds with large grains, large hundreds of grains, high protein content and low grease content can be obtained.

Description

Technical Field

[0001] This invention belongs to the field of molecular genetics technology. More specifically, this invention relates to the application of the soybean GmLEC1 gene and its encoded protein in regulating soybean grain weight, oil or protein content. Background Technology

[0002] Soybean (Glycine max) is the world's most widely cultivated economic crop, providing abundant protein and oil resources for human and animal diets. In recent years, with the continuous improvement of people's living standards, my country's demand for soybeans has risen rapidly, and the supply-demand imbalance has become increasingly prominent. Given the limited arable land area per capita, cultivating high-yield and high-quality new varieties has become the main goal of soybean genetic breeding in my country.

[0003] Seeds are the reproductive organs unique to higher plants and a major source of food for crops. Seed size, oil and protein content are important factors affecting soybean yield and quality, controlled by multiple genetic loci and exhibiting coupled correlations. Traditional conventional breeding methods fall short in terms of breeding efficiency and precise improvement of specific traits. Currently, combining conventional breeding with molecular design breeding is the best approach to achieve breakthroughs in high-yield and high-quality soybean breeding. Analyzing the genetic regulatory network coupled between complex traits and identifying key regulatory units is of great significance for molecular design breeding.

[0004] Although researchers have conducted extensive studies on candidate pleiotropic genes affecting seed size, oil content, and protein content, the transcriptional regulatory networks of many key genes remain unclear. Therefore, identifying key genes that synergistically regulate seed size, oil content, and protein content and elucidating their molecular mechanisms is of practical significance for improving soybean quality and also provides important genetic resources and theoretical guidance for breeding high-yield and high-quality soybean varieties. Summary of the Invention

[0005] Based on this, the purpose of this invention is to provide an application of the soybean GmLEC1 gene and its encoded protein, which can be used to increase soybean grain weight and protein content, and reduce soybean oil content.

[0006] The technical solutions for achieving the above-mentioned objectives include the following.

[0007] In a first aspect, the present invention provides the application of the soybean GmLEC1 gene in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 or SEQ ID NO:2.

[0008] In a second aspect, the present invention provides the application of a protein encoded by the soybean GmLEC1 gene in regulating soybean grain weight, oil content, or protein content, or in the breeding of high-yielding and high-quality soybean varieties, wherein the amino acid sequence of the encoded protein is shown in SEQ ID NO:3 or SEQ ID NO:4.

[0009] In a third aspect, the present invention provides the application of a soybean GmLEC1 gene dual knockout vector in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0010] In a fourth aspect, the present invention provides a method for increasing soybean grain weight, comprising the following steps: constructing a soybean GmLEC1 gene dual knockout vector to cause the soybean GmLEC1 gene to lose the function of the encoded protein; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0011] In a fifth aspect, the present invention provides a method for increasing the protein content of soybeans, comprising the following steps: constructing a double knockout vector of the soybean GmLEC1 gene to cause the protein encoded by the soybean GmLEC1 gene to lose its function; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0012] In a sixth aspect, the present invention provides a method for reducing the oil content of soybeans, comprising the following steps: constructing a double knockout vector of the soybean GmLEC1 gene to cause the protein encoded by the soybean GmLEC1 gene to lose its function; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0013] In this invention, after studying the GmLEC1 gene cloned from soybean, it was found that when a dual knockout vector simultaneously knocking out GmLEC1a (CDS sequence as shown in SEQ ID NO:1) and GmLEC1b (CDS sequence as shown in SEQ ID NO:2) was transferred into Tianlong No. 1 cultivated soybean, soybean cells regenerated into plants. Starting from the T0 generation, homozygous mutant plants resistant to herbicides were screened. The T2 generation homozygous mutant plants showed significantly increased soybean grain weight and 100-grain weight, accompanied by a significant decrease in oil content and a significant increase in protein content. Therefore, the GmLEC1 gene is a key regulatory gene for soybean grain weight, oil content, and protein content.

[0014] This invention reveals for the first time the biological function of the soybean GmLEC1 gene in regulating soybean seed size, 100-seed weight, oil content, and protein content, providing valuable gene resources and theoretical basis for crop breeding. The soybean GmLEC1 gene can be widely applied to high-yield and high-quality soybean breeding, resulting in seeds with large seeds, heavy 100-seed weight, high protein content, and low oil content. Attached Figure Description

[0015] Figure 1 These are the two editing types of the GmLEC1 gene double knockout mutants (Gmlec1-1 and Gmlec1-2) in Example 3 of this invention.

[0016] Figure 2 This is the seed phenotype of Tianlong No. 1 soybean and T2 generation GmLEC1 gene double knockout mutant homozygous lines (Gmlec1-1 and Gmlec1-2) in Example 3 of the present invention.

[0017] Figure 3 The 100-seed weight, oil content, and protein content of Tianlong No. 1 soybean and T2 generation GmLEC1 gene double knockout mutant homozygous lines (Gmlec1-1 and Gmlec1-2) in Example 3 of this invention are shown. Detailed Implementation

[0018] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0019] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0020] Unless otherwise specified, experimental methods in the following examples were performed under standard conditions, such as those described in the fourth edition of *Molecular Cloning: A Laboratory Manual*, edited by Green and Sambrook, published in 2013, or under conditions recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products.

[0021] In some embodiments of the present invention, the application of the soybean GmLEC1 gene in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties is disclosed, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 or SEQ ID NO:2.

[0022] SEQ ID NO:1(672bp)

[0023] ATGGAAACTGGAGGCTTTCATGGCTACCGCAAGCTCCCCAACACAACCTC

[0024] TGGGTTGAAGCTGTCAGTGTCAGACATGAACATGAACATGAGGCAGCAGC

[0025] AGGTAGCATCATCAGATCAGAACTGCAGCAACCACAGTGCAGCAGGAGAG

[0026] GAGAACGAATGCACGGTGAGGGAGCAAGACAGGTTCATGCCAATCGCTAA

[0027] CGTGATACGGATCATGCGCAAGATTCTCCCTCCACACGCAAAAATCTCCGA

[0028] TGATGCAAAGGAGACAATCCAAGAGTGCGTGTCGGAGTACATCAGCTTCA

[0029] TCACCGGGGAGGCCAACGAGCGTTGCCAGAGGGAGCAGCGCAAGACCAT

[0030] AACCGCAGAGGACGTGCTTTGGGCAATGAGTAAGCTTGGATTCGACGACT

[0031] ACATCGAACCGTTAACCATGTACCTTCACCGCTACCGTGAGCTGGAGGGTG

[0032] ACCGCACCTCTATGAGGGGTGAACCGCTCGGGAAGAGGACTGTGGAATAT

[0033] GCCACGCTTGCTACTGCTTTTGTGCCGCCACCCTTTCATCACCACAATGGCT

[0034] ACTTTGGTGCTGCCATGCCCATGGGGACTTACGTTAGGGAAACGCCACCA

[0035] AATGCTGCGTCATCTCATCACCATCATGGAATCTCCAATGCTCATGAACCAA

[0036] ATGCTCGCTCCATATAA

[0037] SEQ ID NO:2(681bp)

[0038] ATGGAAACTGGAGGCTTTCACGGCTACCGCAAGCTCCCCAACACCACCGC

[0039] TGGGTTGAAGCTGTCAGTGTCAGACATGAACATGAGGCAGCAGGTAGCAT

[0040] CATCAGATCACAGTGCAGCCACAGGAGAGAGAACGAATGCACGGTGAG

[0041] GGAGCAAGACAGGTTCATGCCAATCGCCAACGTGATTAGGATCATGCGCA

[0042] AGATTCTCCCTCCACACGCAAAAATCTCGGACGATGCAAAAGAAACAATC

[0043] CAAGAGTGCGTGTCTGAGTACATCAGCTTCATCACAGGTGAGGCGAACGA

[0044] GCGTTGCCAGAGGGAGCAGCGGAAGACCATAACCGCAGAGGACGTGCTT

[0045] TGGGCCATGAGCAAGCTTGGATTCGACGACTACATCGAACCGTTGACCATG

[0046] TACCTTCACCGCTACCGTGAACTTGAGGGTGACCGCACCTCTATGAGGGGT

[0047] GAACCACTCGGGAAGAGGACTGTGGAATACGCCACGCTTGGTGTTGCTAC

[0048] TGCTTTTGTCCCTCCACCCTATCATCACCACAATGGGTACTTTGGTGCTGCC

[0049] ATGCCCATGGGGACTTACGTTAGGGAAGCGCCACCAAATACAGCCTCCTCC

[0050] CATCACCACCACCACCACCACCACCACCATGCTCGTGGAATCTCCAATGCT

[0051] CATGAACCAAATGCTCGCTCCATATAA

[0052] In other embodiments of the present invention, the application of the protein encoded by the soybean GmLEC1 gene in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties is disclosed, wherein the amino acid sequence of the encoded protein is shown in SEQ ID NO:3 or SEQ ID NO:4.

[0053] SEQ ID NO:3(223aa)

[0054] METGGFHGYRKLPNTTSGLKLSVSSDMNMNMRQQQVASSDQNCSNHSAAGE

[0055] ENECTVREQDRFMPIANVIRIMRKILPPHAKISDDAKETIQECVSEYISFITGEA

[0056] NERCQREQRKTITAEDVLWAMSKLGFDDYIEPLTMYLHRYRELEGDRTSMRG

[0057] EPLGKRTVEYATLATAFVPPPFHHHNGYFGAAMPMGTYVRETPPNAASSHHH

[0058] HGISNAHEPNARSI

[0059] SEQ ID NO:4(226aa)

[0060] METGGFHGYRKLPNTTAGLKLSSVSDMNMRQQVASSDHSAATGEENECTVRE

[0061] QDRFMPIANVIRIMRKILPPHAKISDDAKETIQECVSEYISFITGEANERCQREQ

[0062] RKTITAEDVLWAMSKLGFDDYIEPLTMYLHRYRELEGDRTSMRGEPLGKRTV

[0063] EYATLGVATAFVPPPYHHHNGYFGAAMPMGTYVREAPPNTASSHHHHHHHH

[0064] HHARGISNAHEPNARSI

[0065] In other embodiments of the present invention, the application of the soybean GmLEC1 gene dual knockout vector in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties is disclosed, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0066] In some embodiments, the knockout target sequence of the soybean GmLEC1 gene dual knockout vector is shown in SEQ ID NO:9 and SEQ ID NO:10.

[0067] In some embodiments, the soybean GmLEC1 gene dual knockout vector is constructed by the following steps: using the pHLW-gRNA-tRNA vector as a template, SEQ ID NO:11 and SEQ ID NO:12 as primers for PCR amplification and purification, digesting the PCR product and pPTG-gRNA-Cas9 vector plasmid with BsaI enzyme, then performing temperature-dependent cyclic digestion and ligation with T4 DNA ligase, transforming into competent E. coli cells, picking positive clones, and extracting plasmids from the correctly sequenced bacterial cultures to obtain the vector.

[0068] In other embodiments of the present invention, a method for increasing soybean grain weight is disclosed, comprising the following steps: constructing a soybean GmLEC1 gene dual knockout vector to cause the soybean GmLEC1 gene to lose the function of the encoded protein; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0069] In other embodiments of the present invention, a method for increasing soybean protein content is disclosed, comprising the following steps: constructing a soybean GmLEC1 gene dual knockout vector to cause the soybean GmLEC1 gene to lose its protein encoding function; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0070] In other embodiments of the present invention, a method for reducing soybean oil content is disclosed, comprising the following steps: constructing a soybean GmLEC1 gene double knockout vector to cause the soybean GmLEC1 gene to lose the function of the encoded protein; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

[0071] Unless otherwise specified, the methods used in the following examples are conventional methods. Primer synthesis and sequencing can be completed independently or outsourced to a third-party gene company. All vectors and reagents used are commercially available.

[0072] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0073] Example 1: Cloning of the GmLEC1 gene

[0074] This embodiment cloned the GmLEC1a and GmLEC1b genes, including the following steps:

[0075] 1. Using the RNAprep Pure Plant Kit, total RNA was extracted from the seeds of soybean variety Tianlong 1. The cDNA was obtained by reverse transcription using the PrimeScript™ II 1st strand cDNA Synthesis (TaKaRa) kit and used as a cloning template.

[0076] 2. PCR amplification was performed using primers GmLEC1a-F (SEQ ID NO:5) and GmLEC1a-R (SEQ ID NO:6), and GmLEC1b-F (SEQ ID NO:7) and GmLEC1b-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.

[0077] GmLEC1a-F(SEQ ID NO:5):CACACACACCTACCTTATAGC

[0078] GmLEC1a-R(SEQ ID NO:6):TTATATGGAGCGAGCATTTGGT

[0079] GmLEC1b-F(SEQ ID NO:7):CACCTACCTTAATAGCTATGG

[0080] GmLEC1b-R(SEQ ID NO:8):TTATATGGAGCGAGCATTTGGT

[0081] Table 1

[0082] Element volume PCRMix(GenStarcat#A012-101) 20μL template cDNA 1μL Primer GmLEC1a / bF 1μL Primer GmLEC1a / bR 1μL <![CDATA[ddH2O]]> 17μL

[0083] 3. After gel extraction and recovery of the PCR products, TA cloning was performed, followed by transformation into Top10 competent cells. Clones that tested positive by PCR in the bacterial culture were sent to the company for sequencing.

[0084] Sequencing results showed that the CDS length of GmLEC1a was 672bp (SEQ ID NO:1), which encodes 224 amino acids (SEQ ID NO:3); and the CDS length of GmLEC1b was 681bp (SEQ ID NO:2), which encodes 227 amino acids (SEQ ID NO:4).

[0085] Example 2: Construction of the GmLEC1 gene dual knockout vector

[0086] This embodiment constructs a dual knockout vector for the GmLEC1a and GmLEC1b genes, including the following steps:

[0087] 1. Given the high similarity of the nucleotide sequences of GmLEC1a and GmLEC1b, sequence 1 (GGAGAACGAATGCACGGTGAGGG, SEQ ID NO:9) and sequence 2 were selected.

[0088] (TGTACCTTCACCGCTACCGT, SEQ ID NO:10) serves as the common knockout target for both.

[0089] 2. Using the intermediate vector pHLW-gRNA-tRNA as a template, PCR amplification was performed using the forward adapter primer GmLEC1-Cas9-F (SEQ ID NO: 11) and the reverse adapter primer GmLEC1-Cas9-R (SEQ ID NO: 12). 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.

[0090] GmLEC1-Cas9-F (SEQ ID NO:11):

[0091] GGTCTCTTGCAGGAGAACGAATGCACGGTGAGGGGTTTCAGAGCTATGCTGGA

[0092] GmLEC1-Cas9-R (SEQ ID NO:12):

[0093] GGTCTCTAAACACGGTAGCGGTGAAGGTACATGCACCAGCCGGGAATCGA

[0094] Table 2

[0095] Element volume 2×Phanta Max Buffer 20μL Phanta Max Super-Fidelity DNA Polymerase 1μL dNTP Mix 1μL template pHLW-gRNA-tRNA plasmid 1μL Primer GmLEC1-Cas9-F 1μL Primer GmLEC1-Cas9-R 1μL <![CDATA[ddH2O]]> 15μL

[0096] 3. Purify the PCR products according to the kit instructions.

[0097] 4. Cyclic enzyme digestion and ligation

[0098] Add approximately 20–70 ng of PCR product and approximately 80–100 ng of pPTG-gRNA-Cas9 vector plasmid to a 15 μL reaction system, and digest with 10 U BsaI at 37°C for 15 min. Then add 0.5 μL of 10×NEB T4 DNA ligase buffer and 0.1 μL of 35 U ligase, and perform 12 cycles of variable-temperature cyclic digestion and ligation: 37°C for 2 min, 10°C for 3 min, 20°C for 5 min; and finally 37°C for 2 h.

[0099] 5. Transformation of competent states and identification

[0100] The ligation product was transformed into top10 competent E. coli cells and plated for culture. Positive clones were picked and expanded in LB broth containing the corresponding antibiotics. PCR identification and sequencing verification were performed using primers SP-F (SEQ ID NO:13) and SP-R (SEQ ID NO:14), thereby obtaining the dual knockout vectors of the GmLEC1a and GmLEC1b genes.

[0101] SP-F (SEQ ID NO:13):GTCGTGTCCACATGTTGACCGG

[0102] SP-R(SEQ ID NO:14):CCCGACATAGATGCAATAACTTC

[0103] Example 3: Agrobacterium-mediated transformation of soybeans

[0104] Includes the following steps:

[0105] 1. Obtaining soybean explants

[0106] Tianlong No. 1 soybean seeds with smooth surfaces, no cracks, no disease spots, and no mold were sterilized with chlorine for 10–14 hours, and then cultured in the dark on MS basal medium. After germination, the two cotyledons were cut along the midline in a clean bench, the two original leaf buds were removed, and a wound about 3 mm long was made at the junction of the cotyledon and hypocotyl.

[0107] 2. Soybean genetic transformation

[0108] Using Agrobacterium-mediated genetic transformation, the EHA105 strain carrying the dual knockout vector of GmLEC1a and GmLEC1b genes from Example 2 was infected with cotyledon wounds, and transgenic plants were obtained after induction culture.

[0109] 3. Screening for heritable transgenic plants

[0110] Transgenic seedlings of generation T0 were screened by spraying with 160 mg / L glufosinate (Basta), and positive plants were obtained by PCR identification. Subsequently, generation T1 plants were obtained through propagation. Total DNA was extracted from the leaves of generation T2 transgenic plants. The F-terminal primer (GCAGCAGGTAGCATCATCAGA, SEQ ID NO:15) and R-terminal primer (GCAGCATTTGGTGGCGTTTC, SEQ ID NO:16) were used to amplify the sequences before and after the target site. Sequencing verification yielded two homozygous lines, Gmlec1-1 and Gmlec1-2, with specific editing methods as follows: Figure 1 As shown.

[0111] 4. Quality assessment of transgenic plants

[0112] Phenotypic observations were performed on soybean seeds of two different editing types, and their 100-seed weight, oil content, and protein content were determined. The results are as follows: Figure 2 and Figure 3 As shown.

[0113] Figure 2 and Figure 3 The results showed that, compared with the cultivated soybean variety Tianlong 1 (TL1), both edited types (Gmlec1-1 and Gmlec1-2) of soybeans exhibited larger grains and a significantly higher 100-grain weight. Simultaneously, the protein content in the grains was significantly increased, while the oil content was significantly decreased.

[0114] The above results indicate that the GmLEC1 gene not only plays an important role in regulating grain size and weight, but also participates in the regulation of oil and protein accumulation.

[0115] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. Application of the soybean GmLEC1 gene in regulating soybean grain weight, oil content or protein content, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 or SEQ ID NO:

2.

2. Application of the soybean GmLEC1 gene in the breeding of high-yield and high-quality soybean varieties, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 or SEQ ID NO:

2.

3. The application of the protein encoded by the soybean GmLEC1 gene in regulating soybean grain weight, oil content or protein content, wherein the amino acid sequence of the encoded protein is shown in SEQ ID NO:3 or SEQ ID NO:

4.

4. Application of the protein encoded by the soybean GmLEC1 gene in the breeding of high-yield and high-quality soybean varieties, wherein the amino acid sequence of the encoded protein is shown in SEQ ID NO:3 or SEQ ID NO:

4.

5. Application of the soybean GmLEC1 gene dual knockout vector in regulating soybean grain weight, oil content or protein content, or in the breeding of high-yield and high-quality soybean varieties, wherein the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:

2.

6. The application according to claim 5, characterized in that, The knockout target sequence of the soybean GmLEC1 gene dual knockout vector is shown in SEQ ID NO:9 and SEQ ID NO:

10.

7. The application according to claim 5 or 6, characterized in that, The soybean GmLEC1 gene dual knockout vector was constructed through the following steps: using the pHLW-gRNA-tRNA vector as a template, SEQ ID NO:11 and SEQ ID NO:12 as primers for PCR amplification and purification, the PCR amplification product and pPTG-gRNA-Cas9 vector plasmid were digested with BsaI enzyme, followed by temperature-dependent cyclic digestion and ligation with T4 DNA ligase, and then transformed into competent E. coli cells. Positive clones were selected, and plasmids were extracted from the correctly sequenced bacterial cultures.

8. A method for increasing soybean grain weight, characterized in that, Includes the following steps: A dual knockout vector for the soybean GmLEC1 gene was constructed to render the protein encoded by the soybean GmLEC1 gene inoperable; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:

2.

9. A method for increasing the protein content of soybeans, characterized in that, Includes the following steps: A dual knockout vector for the soybean GmLEC1 gene was constructed to render the protein encoded by the soybean GmLEC1 gene ineffective; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:

2.

10. A method for reducing the oil content of soybeans, characterized in that, Includes the following steps: A dual knockout vector for the soybean GmLEC1 gene was constructed to render the protein encoded by the soybean GmLEC1 gene ineffective; the CDS sequence of the soybean GmLEC1 gene is shown in SEQ ID NO:1 and SEQ ID NO:2.

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