CN118652906A - Cloning and application of gene for regulating size of soybean seeds - Google Patents

Abstract

The invention provides a soybean grain weight related protein, a coding gene and application thereof, wherein the coding region sequence of the gene is shown as SEQ ID NO. 1 or a homologous sequence thereof. The invention also provides a method of growing a yield-enhanced soybean comprising transforming GmSGF gene or a vector or host cell comprising GmSGF gene into a soybean plant cell or tissue and growing to obtain a yield-enhanced soybean plant. Also disclosed is the use of GmSGF gene or a vector or host cell comprising GmSGF gene for breeding soybeans with increased yield. The invention has great theoretical and application value for soybean breeding and related application research.

Description

Cloning and application of gene for regulating size of soybean seeds

Technical Field

The invention belongs to the field of biotechnology, and particularly relates to cloning and application of a gene for regulating and controlling the size of soybean seeds, and more particularly relates to cloning and application of a gene for regulating and controlling the weight of soybean seeds.

Background

Soybean (Glycine max L. Merr.) is one of the world's important oil and commercial crops, and is also a major source of human premium and feed proteins. Soybean origin China. The main reason is that the soybean production per unit in China is low, so that the soybean production per unit in China is improved.

The yield of soybeans depends to a large extent on the grain weight, i.e., grain size (including grain length, grain width, and grain thickness). However, so far gene clones for regulating soybean seed size are still relatively limited, and the genetic basis and specific regulatory mechanisms in soybean germplasm resource variation are not clear. Therefore, deep excavation of molecular genetic mechanism for regulating and controlling soybean seed size has important significance for improving soybean yield.

Disclosure of Invention

The invention aims to provide a soybean grain weight related protein, a coding gene and application thereof, and provides theoretical basis and gene resources for subsequent molecular assisted breeding and molecular design breeding.

Wherein, in the specific embodiment of the invention, the cDNA sequence of the soybean grain-weight related GmSGF gene is shown as SEQ ID NO.1, and in the specific embodiment of the invention, the amino acid sequence of the GmSGF gene is shown as SEQ ID NO. 2. In a specific embodiment of the present invention, the genomic DNA (gDNA) sequence of the GmSGF gene is shown as SEQ ID NO. 3.

In the present invention, a recombinant vector containing a target gene can be constructed using an existing plant expression vector. The plant expression vector comprises a binary agrobacterium vector, a vector which can be used for plant microprojectile bombardment and the like. To facilitate identification and selection of transgenic plant cells or plants, the plant expression vectors used may be processed, for example by adding genes encoding enzymes or luminescent compounds which produce a color change, antibiotic markers or chemical resistance marker genes which are expressed in the plants, etc. From the safety of transgenic plants, transformed plants can be screened directly in stress without adding any selectable marker gene. Enhancers may also be included in the plant expression vector to increase the expression of the inserted nucleotide fragment.

In order to achieve the above object, the present invention also provides a method for obtaining transgenic soybean, which comprises introducing the nucleic acid or the vector or host cell comprising the gene into a soybean of interest to obtain a transgenic soybean exhibiting increased grain size as compared with the soybean of interest.

Among them, the method of introducing the soybean of interest may be by transforming plant cells or tissues using conventional biological methods such as Ti plasmid, ri plasmid, plant viral vector, direct DNA transformation, microinjection, electric conduction, agrobacterium mediation, etc., and culturing the transformed soybean cells or tissues into plants.

In order to achieve the above object, the present invention also provides the use of a protein as defined above or a nucleic acid as defined above or a vector or host cell comprising the above gene in soybean genetic engineering.

Among them, the soybean genetic engineering is preferably soybean genetic engineering for the purpose of improving soybean yield.

The soybean grain weight related protein and the nucleic acid encoding the same provided by the invention have the functions of regulating and controlling soybean grain weight such as hundred grain weight for the first time, and the phenotypic analysis verification of transgenic plants and wild plants shows that the soybean grain weight related protein can increase the grain weight of soybean transgenic plants. The invention has great theoretical and application value for soybean high-yield breeding and related application research.

Specifically, the invention provides the following technical scheme:

In one aspect, the invention provides an isolated gene for regulating soybean grain weight, wherein the coding region sequence of the gene is shown as SEQ ID NO. 1 or a homologous sequence thereof.

In some embodiments, the genomic sequence (gDNA) of the gene is as shown in SEQ ID NO.3 or a homologous sequence thereof.

In another aspect, the invention provides a protein for regulating soybean grain weight, wherein the amino acid sequence of the protein is shown as SEQ ID NO. 2 or a homologous sequence thereof.

In another aspect, the invention provides an expression vector comprising a gene as described above or a nucleotide sequence encoding a protein as described above.

In some embodiments, the expression vector has an antibiotic marker or an anti-chemical agent marker.

In some embodiments, the antibiotic marker is selected from the group consisting of ampicillin, chloramphenicol, kanamycin, neomycin, rifampicin, spectinomycin, hygromycin, streptomycin, and tetracycline, and the anti-chemical agent marker is, for example, an anti-herbicide marker.

In another aspect, the invention provides a host cell comprising an expression vector as described above.

In another aspect, the invention provides a method of growing soybean with increased yield comprising transforming a gene as described above or a nucleotide sequence encoding a protein as described above or an expression vector or host cell as described above into a soybean plant cell or tissue and growing to obtain a soybean plant with increased yield.

In some embodiments, the vector is a plant expression vector, including binary agrobacterium vectors and/or vectors useful for plant microprojectile bombardment.

In some embodiments, the host cell is selected from an e.coli cell, an agrobacterium cell, or a plant cell.

In some embodiments, the agrobacterium is selected from EHA105, EHA101, and GV3101

In another aspect, the invention provides the use of a gene as described above or a nucleotide sequence encoding a protein as described above or an expression vector or host cell as described above for breeding soybeans with increased yield.

In some embodiments, the increase in yield is manifested by an increase in grain width, an increase in grain length, an increase in grain thickness, a grain enlargement, an increase in hundred grain weight, and/or an increase in individual yield.

In another aspect, the invention provides a method of growing a transgenic plant with increased yield, which comprises introducing a gene as described above or a nucleotide sequence encoding a protein as described above or an expression vector or host cell as described above into a plant cell or tissue of interest to obtain a transgenic plant with increased yield compared to the plant of interest, which plant is a leguminous plant, preferably soybean.

Drawings

FIG. 1 shows the results of the expression level analysis of wild-type and overexpressing transgenic plants in GmSGF (version W82 Gene number: glyma.11G16846), wherein the ordinate shows the expression level of GmSGF11 and the abscissa shows the line number. Significance analysis was tested using Student's t (< 0.05, <0.001, < p).

Figure 2 shows the phenotypic statistics of wild type and transgenic plants overexpressing GmSGF 11. Wherein A: the grain width phenotype of the wild type and the transgenic plants overexpressing GmSGF are compared, the scale bar being 1cm. B: the grain length phenotype of wild type and transgenic plants overexpressing GmSGF11 were compared, with a scale bar of 1cm. C: the grain thickness phenotype of the wild type and the overexpressing GmSGF11 transgenic plants was compared, the scale bar being 1cm. D: statistical results of grain width of wild type and overexpressing GmSGF11 transgenic plants. E: statistical results of grain length of wild type and overexpressing GmSGF11 transgenic plants. F: statistical results of grain thickness for wild type and overexpressing GmSGF11 transgenic plants. G: statistics of hundred grain weights of wild type and overexpressing GmSGF11 transgenic plants. H: statistics of individual yields of wild type and overexpressing GmSGF11 transgenic plants. Significance analysis was tested using Student's t (< 0.001).

Detailed Description

The present invention will be further described in detail below with reference to specific embodiments and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the present invention more apparent.

The following examples are provided to facilitate a better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples, unless otherwise specified, were either conventional or selected according to the commercial specifications. The test materials used in the examples described below, unless otherwise specified, are all commercially available conventional biochemical reagents. The quantitative tests in the following examples were all set up in triplicate and the results averaged.

In the examples described below, the transformation recipient was dongnong 50 (DN 50), i.e. wild type, which is a black-lobed cross-sectional variety (black bean 2007022) available from the market. Agrobacterium strain EHA101 was purchased from China center for plasmid vector strain cell gene collection (Biovector Science Lab, inc).

Consumables such as the digestion recovery kit were purchased from NEW ENGLAND Biolabs and Tiangen Biotechnology (Beijing) Inc.

PFGC5941 vector is available from Bekyphosis bordetella biotechnology Co., ltd., product number vt-3034, and the details of the vector and method of use are given in the product description, reference being made to ：Kerschen A,Napoli C A,Jorgensen R A,et al.Effectiveness of RNA interference in transgenic plants[J].FEBS letters,2004,566(1-3):223-228.

Example 1GmSGF discovery of protein and Gene encoding same

Based on a large number of sequence analyses and functional verification, a protein is found from soybean variety Williams 82, and is named GmSGF protein, and the amino acid of the protein is shown as SEQ ID NO in a sequence table: 2, the gene encoding GmSGF11 protein is named GmSGF gene, and the genome sequence of the gene is shown as SEQ ID NO:3, the cDNA sequence of which is shown as SEQ ID NO: 1.

Functional verification of GmSGF protein from example 2

1. Construction of recombinant plasmids

1. Separating seeds of the soybean variety Williams 82R 4 stage (Cheng Jia stage) from plants, and extracting RNA to obtain seed RNA of the soybean variety Williams 82.

2. The total RNA obtained in step 1 was reverse transcribed into cDNA using a reverse transcription kit (full gold). And (3) taking cDNA as a template, and carrying out PCR amplification by using a primer pair consisting of F1 and R1 to obtain a PCR amplification product.

F1：5’-ATGTCTGTTCCTAATGAGGAA-3’(SEQ ID NO：4)；

R1：5’-TCATGACTCCAGTGTCCCATTTG-3’(SEQ ID NO：5)。

3. The PFGC5941,5941 vector was digested with the restriction enzymes AscI and XmaI to recover a vector backbone of about 9992 bp.

4. And (3) amplifying the PCR product in the step (2) by using a primer consisting of F2 and R2 to obtain a PCR amplification product (2).

F2：5’-cattacaattacatttacaattacATGTCTGTTCCTAATGAGGAA-3’(SEQ ID NO：6)；

R2：5’-caggactctagggactagtcccgggTCATGACTCCAGTGTCCCATTTG-3’(SEQ ID NO：7)。

5. And (3) connecting the PCR product in the step (4) with the vector skeleton in the step (3) to obtain the recombinant plasmid. According to the sequencing result, the recombinant plasmid A is structurally described as follows: the sequence table is inserted between the AscI and Xma I enzyme cutting sites of PFGC5941 vector, and the sequence table is shown as SEQ ID NO:1, and a double-stranded DNA molecule shown in 1.

2. GmSGF11 acquisition of transgenic plants overexpressing GmSGF

1. The recombinant plasmid was introduced into Agrobacterium strain EHA101 (purchased from China center for type culture Collection of plasmid vector strains) to obtain recombinant Agrobacterium.

2. The recombinant agrobacterium obtained in step 1 is transformed into a recipient plant DN50 by adopting a cotyledonary node transformation method (see Margie M.P.et al 2004Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant.Euphytica 136:167–179) for specific operation methods, and T0 generation seeds are harvested:

(1) Sterilizing soybean seeds: selecting full soybean seeds without scars on seed coats, subpackaging the soybean seeds, placing the soybean seeds into a culture dish, opening the cover of the culture dish, placing the culture dish into a dryer, sterilizing the seeds by a chlorine fumigation method, placing a small beaker into the dryer, adding 100mL of sodium hypochlorite, then adding 5mL of concentrated hydrochloric acid, quickly covering a cover, sterilizing for 10-14h, taking out the soybeans from the dryer after sterilization is completed, and blowing air for more than 30min in an ultra-clean bench to eliminate redundant chlorine;

(2) Swelling soybean seeds: inserting sterilized seed into germination medium (table 1) with umbilicus downward, standing at 23deg.C in dark for 16-18 hr;

(3) Preparing agrobacterium: the bacteria stored at-80℃were removed, added to 5mL of YEB medium (Table 2) containing the corresponding resistance, cultured at 28℃at 250rpm for 24 hours for resuscitation, and then 200. Mu.L of the resuscitated bacterial liquid was aspirated and added to 200mL of YEB medium containing the corresponding resistance, cultured at 28℃at 250rpm until OD ₆₀₀ = 0.6, and the bacterial liquid was collected. Centrifuging at 22deg.C at 5,000rpm for 10min, and then re-suspending the cells with liquid dip-dyeing medium (Table 3) to OD ₆₀₀ =0.5, culturing at 22deg.C at 70rpm for 0.5 hr;

(4) Preparation of explants: taking out the seed coat of the swelled seed, cutting off the part of the hypocotyl away from the cotyledon by using a surgical knife to ensure that the residual hypocotyl is about 2-3mm, dividing the soybean seed into two parts, and removing germs by using a small iron brush to obtain the prepared explant;

(5) Infection of explants: placing the prepared explant into an agrobacterium suspension, enabling the heavy suspension to permeate the explant, and slightly shaking and culturing for 0.5h;

(6) Co-culturing the explant and agrobacterium: the infected explant is sucked off with excess bacterial liquid on sterile filter paper, a piece of sterile filter paper is placed on a co-culture medium (table 4), the sucked-off explant is placed on the co-culture medium and is cultivated in the dark for 3-5d at the temperature of 23 ℃;

(7) Induction culture of callus: after the co-culture of the explant and the agrobacterium, transferring the explant into a callus induction medium (table 5), at this time, slightly inserting the explant into the medium at an inclination of 45 degrees and at the bottom, culturing for 14d at 23 ℃ for 16h under illumination/8 h darkness;

(8) And (3) bud induction culture: the induced callus is excised from the explant and placed in a bud induction medium (table 6) at 23 ℃ for 16h light/8 h darkness for 14d, and transferred into a new bud induction medium every 14 d;

(9) Bud elongation culture: transferring the explant with the induced new bud into a bud elongation culture medium (table 7), and transferring the explant into the new bud elongation culture medium every 2 times until the bud is elongated to more than 3 cm;

(10) Rooting culture: the elongated shoots were cut out and inserted into rooting medium (Table 8) until the roots were over 3cm long, then the seedlings were acclimatized, transplanted into a greenhouse, at 24℃under 16h light/8 h darkness until maturation, during which time Basta and PCR were sprayed to identify positive lines.

TABLE 1 germination medium

GM (germination medium)	1L
		B5 (major, minor, ferric salt)	3.1g
Sucrose (Sucrose)	20g
		pH	5.8
Plant gel (Phytagel)	2.5g
		Sterilization	121℃，20min
B5 vitamins (1000×)	1mL

TABLE 2YEB Medium

TABLE 3 infection liquid

LCCM (infection liquid)	1L
		B5 (major, minor, ferric salt)	0.31g
Sucrose (Sucrose)	30g
		MES (fatty acid methyl ester sulfonate)	3.9g
pH	5.4
		Sterilization	121℃，20min
B5 vitamins (1000×)	1mL
		GA3(1mg/mL)	0.25mL
6-BA(1mg/mL)	1.67mL
		DTT(100mg/mL)	1.8mL
As(100mg/mL)	0.8mL
		Silwet	20μL/100mL

TABLE 4 Co-culture Medium

TABLE 5 callus induction medium

TABLE 6 bud induction Medium

TABLE 7 bud elongation Medium

TABLE 8 rooting medium

Transgenic phenotype statistics of example 3GmSGF11

1. Identification of Gene expression level

The receptor lines DN50, gmSGF11 over-expressed transgenic lines were identified as follows:

(1) Seeds of DN50 and GmSGF11 over-expressed transgene R4 stage are separated from plants, total RNA is extracted and reverse transcribed into cDNA.

(2) Using the cDNA extracted in the step (1) as a template, identifying GmSGF gene expression levels using a primer pair consisting of F4 and R4, and identifying reference genes (Actin gene expression levels) using F3 and R3.

F3：5’-CGGTGGTTCTATCTTGGCATC-3’(SEQ ID NO：8)；

R3：5’-GTCTTTCGCTTCAATAACCCTA-3’(SEQ ID NO：9)。

F4：5’-TGTGGTCGATCCATCATGGCTG-3’(SEQ ID NO：10)；

R4：5’-CCAACCGATTCATGCTATTGGTA-3’(SEQ ID NO：11)。

The result of quantitative PCR amplification of the cDNA as a template using each specific primer pair shows the amount of GmSGF gene expressed in different materials as shown in FIG. 1. As can be seen from FIG. 1, the gene expression levels of GmSGF were significantly higher than the control DN50 in the three over-expressed lines GmSGF-OE-1, gmSGF-OE-2 and GmSGF-OE-3.

2. Seed size, grain weight and individual yield comparison of recipient lines DN50, gmSGF11 over-expressed transgenic lines

Seeds of the recipient lines DN50, gmSGF11 over-expressed transgenic lines were subjected to statistics of grain size and hundred grain weight and individual yield, and the results are shown in FIG. 2. As can be seen from FIG. 2, gmSGF11 over-expressed transgenic lines showed significantly larger kernels than DN50 (embodied as 9.7% increase in grain width, 9.1% increase in grain length, 9.7% increase in grain thickness), indicating that GmSGF gene is a key gene regulating soybean yield, and over-expression of GmSGF gene in soybean plants can promote soybean kernels to become larger, increase in hundred grain weight (19% increase), and thus increase individual yield of soybean (30% increase).

Sequence listing

SEQ ID NO. 1GmSGF11 546bp cDNA Glycine max (Soybean)

ATGTCTGTTCCTAATGAGGAAAACTTGTCGTCACATTCCCAGCTTTCTTCTCATTTTTTCTTGGATCTCCTTGATTCCATCATAGTTGATGTGGCATCAGAGTGTCACAGAGTAGCAAGGCTGGGGCTTGATTCTAATTTGGAAGAAGAAGATGAAGAATTGAAGCTATCGGCACAAGCCAGGGTTAGGGTGGCTGATCCTAGTAACAGTAATGAAGCAAATGGCAAGTATGTGGTTGACATATTTGGACAAACCCATCCTCCTGTGGCAAATGAAATATTTGATTGCATGAATTGTGGTCGATCCATCATGGCTGGGAGGTTTGCTCCACATTTGGAGAAGTGCATGGGAAAGGGTAGGAAGGCACGTCTGAAAGTGACAAGAAGCAGCACAGCCGCGCAGAACCGGTATTCACGAGGCAGTCCTAGTCCTGGTTCTACATATTCTCCATATTCAAATTACTCTACCAATAGCATGAATCGGTTGGCAAATGGAACCTCCACTTTTGCAGGTGAGGAGCACTCAAATGGGACACTGGAGTCATGA

SEQ ID NO. 2GmSGF11 182aa protein Glycine max (soybean)

MSVPNEENLSSHSQLSSHFFLDLLDSIIVDVASECHRVARLGLDSNLEEEDEELKLSAQARVR VADPSNSNEANGKYVVDIFGQTHPPVANEIFDCMNCGRSIMAGRFAPHLEKCMGKGRKARLKVT RSSTAAQNRYSRGSPSPGSTYSPYSNYSTNSMNRLANGTSTFAGEEHSNGTLES*

SEQ ID NO. 3GmSGF11 3795bp gDNAGlycine max (Soybean)

AAATAGTTTGAACTCATATTTGAGTATTTACCCTAAATTTTAGTTGTATATGGGTCTTTTATTTGGAGGCAGGGTCAAATAACACAACCCTCATGGGAGGTGCCAATAGTAGACCCCTAAAATTTTCTTTTCTAGCCCAAAAGTTTAGGCTTTTTCGAAATTGTACCCCAGTGGCATTTGAGGGGTTGCTCTGTGAATCACAGAATCTCTGTTAGCACTCTCCTCTCGACGATTCCCTGTGATCTCTCTCTCTCTTTCTCTCTCTTTCTCTCTCAGTCTTCTCTCGCTGTGGTTCAGATTTCAGAGCAGGTTCGTCTCTCATCTCTGCGGTTTAGGTTTCGATTTTTTATTCCGCTAATTTGTGTTTTCTTTCATTAATTTGCATTTCTTTTCTTTTTCTTTTAATTTGCATCACCTAAATTTTGTTACTTTATGAAACTGAAGAACATGAAAGACGGAAAAACAACTTTTTATGAACCAATCATATTTGGAGGGGAAGGATCCTTAAGCCAGTGGTTTGGTTCATAATAGGGTTGAAATTAACAGCTCTTTTGTTTTTGAGGATAGGCTTTACCAATATTGTAGATATCAACATTGTTTCCTTTTGTTATTATTATCTTTTTTTTTTGGTTCTTTTGTTCTTGTCTTTAATTTATACTTCCATGAAGCTTTTTGTGGACGTTTCTGACTTTCACCAGTTATTGGTGTAAATTAGGATCCAAAAGCCTTGCTTGGTCAAGACTCATTCATGTCTGTTCCTAATGAGGAAAACTTGTCGTCACATTCCCAGGTGTGCTCTTCAATTCTGCAAAGACATGCTTTTATGAGTGCAGCAATAATTAAATATTAGATTTTGATGTTAATTCCTAGTACATTTCAAAGTCTATTTAAACTTTTGTTGCAAACCATGCTTGGGTGCATTTCAGCTTTCAATTTTGCATTCTCATAAGACTTGATCCAGGGATAAGTTTTGTGATTGCATTAGTTGTTTTAATCTAGAATTGTCTTTTGGAAAAAATGAGTATTGGTTGAACTCCATTTGTCTTTCATTAATATGTCTACGACTTTGTGATCATTGTTCTTTTCCTGTTTTTTATCCCTGCCTCCTCCCCCTCTTAAAATTAACTTCTGAAAAGATGCTGTTGAAACGTAACATGCCTAAAATTGGTACTTTTTCCCTTGTGGCTATGATCGACAGTGAAAAAGAAGATTAAGATCAATAATGAAGGGAGCAATAATTTAATAAGGCAAGAAGAATTGTGAATTCATTAGATTGTTTCTCATTCATTTGATTGTACTGCCGAACATTGCTTTTCCTGCAAGTTGTTGGAGCAGATTATTCTTTGGGATATAACATGATAAAATTTCAGTGGTTGCAAATGTATATTTTAATACTCCATTTATAAAGGATAATAATACCATGGTGAATATATTATGATAATTATCAAATAAATTGAATTGTTTAAGCCTTAGTCCCTTACACAACCCTTATATACTTTTGTGACTTATGCACTTGTCCTTCATACGCACTTTTGCTGCAAGTCAATACTTTGTACCTGTAGATACTAGATATCCTTGATTAGGTGGAGTTTCTACTTTCACTGTTTGTCTAGAGTCTAGAGAAATATGAAAATCCATGTCCCAGCAGAGATGCCATGATTTCAATTTTTATCATCAATGGGTATCATCATTCAGTTGTTTTTATTTTTTTGCTTTTCATTTTTTTCCTTTTCAGAGAAAGATGGTTCAGTGATGCATGAAGTGGAAAGTCTATCTTGAACTGCCAACCTTTCTATCTGCTCTCATATCTTTCTTTTAGAATGAGGATGGAGGGAATTGTAAATGCCACTCTTCAACCCTTATTCTGTTACCTTAAATAAATGAATAACACTAAATGTTAGATGCTAGTCTCAGTAGTATACCTTCTATCATATTTGTATTTCTTTGTATTGTGGAATAACTCTCACATGCATGTGCCTAAAGTAATTGGTTATGGGCCTGTTTGGATGTACTACTGTAGAAGATTTTATAGGAAAAAAACTTATTCCATATGTTTTCCTACTATATGTGTTCCATATGCATTACTGTTTAGATGCTAGTCTCACTCTCAGTGAATTGAAACATTCAGAATAATACATATGTTTTCCTACTATATATAATATATTTATATGGTTATTTGCTTCCATTGGGGTGTTTTGCAGCTTTCTTCTCATTTTTTCTTGGATCTCCTTGATTCCATCATAGTTGATGTGGCATCAGAGTGTCACAGAGTAGCAAGGCTGGGGCTTGATTCTAATTTGGAAGAAGAAGATGAAGAATTGAAGCTATCGGCACAAGCCAGGGTTAGGGTGGCTGATCCTAGTAACAGTAATGAAGCAAATGGCAAGTATGTGGTTGACATATTTGGACAAACCCATCCTCCTGTGGCAAATGAAATATTTGATTGCATGAATTGTGGTCGATCCATCATGGCTGGGAGGTTTGCTCCACATTTGGAGAAGTGCATGGGAAAGGTTATTTTTTTACCTTCTAGAATTGTTTATTTTTTAATAAAATATTTACTCATTATGCCCATAACGTTAGTCACTATCATAAATTTCCAATGAATGCTAGCAACACATTCTCTAACACACTCCTCATACACTATTGTTCTCAATTTATTGAAAACTACAAAACCATGAATGAAACTCATTAAATAAGTGAGACCTGGATGATTTTCAATCAATTTCAGCCAATAGTAGAGACTAGAGAGTATGTTAGAGAGTGTATTGCTAACATTTCTCATTTAAAATATATCTATTCATGTTATCACATGGCATTTGTGGAGAAGACTATAAAAACTTTAGATAGAGGGTAAGGGTAGCCCAATCATTAAAGGGAGATCAAGAAAAACTATAAGTGAAACTATTGAGAAGGATTTAGAGATTAATGATTTATTTATAGACACATTTTACTATAGGATACTATGGTGTTGCTTGATCCATGTAGTCGACTTAATGTGGGGGGGGGGGGGGGGGGGAACTTGTGGTTATTGATAACTCAGTGAAAATAATATTCTCAATCTTCATTCATGCATGTAATTGAGCTTTTTATTTTTTCACTTATGTGTCATTTTCCTATGTAATTGAGCTTTATATTTTTTCACTTATGTGTCATTTTCCTATTATGGAAATGCTTGCATGCTTCATTTCAGGGTAGGAAGGCACGTCTGAAAGTGACAAGAAGCAGCACAGCCGCGCAGAACCGGTATTCACGAGGCAGTCCTAGTCCTGGTTCTACATATTCTCCATATTCAAATTACTCTACCAATAGCATGAATCGGTTGGCAAATGGAACCTCCACTTTTGCAGGTGAGGAGCACTCAAATGGGACACTGGAGTCATGAACTAGTTCATGTGTAATGGCTGGAATACAGGATCTCTGTCATACCTTGAACTTGTTTCTTCGCCGGGAGATGTGGCTGAATTTAATTGTGTTTGAAAATAATATTTCAAACATTTTGGACCTAAACATAACAACCATCGTTGCCTACATCCTCGAATTGTTGTCTACCTTAAAAAAAAAAAACTTTGCTTTTTACAGTGAGTTCCATTGTAATTTTATTATGTTTTACTCCTAGTTCCGTTGTAATTTTATTATAATTTTACTATCTGTTAACATATGTATATTGTGTGCAACTATGAACTAGTATTGTTCATGGATGAAAAGATGCTCAGGGTTGAGATGAATAATACACAGACATTTGGGAAAGTGTGTGTGTTAAGATGAAAAAATATGTAGAGTTCGTTTGATTTG

The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the invention, and is not meant to limit the invention thereto, but to limit the invention thereto, and any modifications, equivalents, improvements and equivalents thereof may be made without departing from the spirit and principles of the invention.

Claims (10)

1. The isolated gene for regulating and controlling the grain weight of the soybean is characterized in that the coding region sequence of the gene is shown as SEQ ID NO. 1 or a homologous sequence thereof.

2. The gene according to claim 1, wherein the genomic sequence of the gene is as shown in SEQ ID NO. 3 or a homologous sequence thereof.

3. The protein for regulating and controlling the grain weight of the soybean is characterized in that the amino acid sequence of the protein is shown as SEQ ID NO. 2 or a homologous sequence thereof.

4. Expression vector, characterized in that it comprises a gene according to claim 1 or 2 or a nucleotide sequence encoding a protein according to claim 3.

5. The expression vector according to claim 5, characterized in that it has an antibiotic marker or an anti-chemical marker, optionally selected from the group consisting of ampicillin, chloramphenicol, kanamycin, neomycin, rifampin, spectinomycin, hygromycin, streptomycin and tetracycline, for example an anti-herbicide marker.

6. The expression vector of claim 4 or 5, wherein the vector is a plant expression vector, comprising a binary agrobacterium vector and/or a vector useful for plant microprojectile bombardment.

7. A host cell, characterized in that it comprises an expression vector according to any one of claims 4-6.

8. The host cell of claim 7, wherein the host cell is selected from the group consisting of an escherichia coli cell, an agrobacterium cell, and a plant cell, optionally the agrobacterium is selected from EHA105, EHA101, and GV3101.

9. A method of growing a yield-enhanced soybean comprising transforming an expression vector according to any one of claims 4-6 or a host cell according to claim 7 or 8 into a soybean plant cell or tissue and growing to obtain a yield-enhanced soybean plant, optionally said yield enhancement being manifested by increased grain width, increased grain length, increased grain thickness, increased hundred grain weight and/or increased yield per plant.

10. Use of a gene according to claim 1 or 2 or a nucleotide sequence encoding a protein according to claim 3 or an expression vector according to any one of claims 4-6 or a host cell according to claim 7 or 8 for breeding yield-enhanced soybean, optionally the yield enhancement being manifested by increased grain width, increased grain length, increased grain thickness, increased grain size, increased hundred grain weight and/or increased individual yield.