CN112410309A

CN112410309A - Application of GmAAP protein and GmAAP gene in soybean breeding

Info

Publication number: CN112410309A
Application number: CN202011354096.3A
Authority: CN
Inventors: 张丹; 吕海燕; 杨宇明; 张恒友; 王莉
Original assignee: Henan Agricultural University
Current assignee: Henan Agricultural University
Priority date: 2020-11-27
Filing date: 2020-11-27
Publication date: 2021-02-26
Anticipated expiration: 2040-11-27
Also published as: CN112410309B; NL2027897A; NL2027897B1

Abstract

The invention belongs to the field of plant genetic engineering, and particularly relates to application of a GmAAP protein and a GmAAP gene in soybean breeding. The invention provides an application of GmAAP protein in soybean breeding, wherein the amino acid sequence of the GmAAP protein is shown as SEQ ID NO. 1. The invention can obviously increase the branching number of the soybeans by reducing the expression of the GmAAP gene through a genetic engineering technology, thereby improving the yield of the soybeans. The embodiment shows that the application provided by the invention can effectively improve the branch number of the soybeans, improve the pod number of a single plant and further improve the yield of the soybeans.

Description

Application of GmAAP protein and GmAAP gene in soybean breeding

Technical Field

The invention belongs to the field of plant genetic engineering, and particularly relates to application of a GmAAP protein and a GmAAP gene in soybean breeding.

Background

Nitrogen is one of the essential macro-elements for plant growth, and its deficiency directly affects the yield and quality of crops. In order to ensure the yield and quality of crops, a large amount of nitrogen fertilizer is applied to soil every year, however, due to the low absorption and utilization rate of the nitrogen fertilizer, the excessive application of the nitrogen fertilizer not only has extremely limited contribution to the yield of the crops, but also causes the problems of resource waste, environmental pollution and the like. Therefore, the improvement of the capability of the plant to utilize nitrogen is of great significance to the sustainable development of agricultural production. By utilizing the genetic engineering technology, the absorption, utilization and transport capacity of crops to nitrogen in soil are improved, and the cultivation of new varieties of nitrogen-efficient crops is an effective way for solving the problems.

Plants absorb nitrate ions, ammonium ions or amino acids from soil by using root systems to obtain nitrogen, and the absorption and transportation of nitrogen are mainly completed by transporters such as ammonium transport protein (AMT), nitrate transport protein (NRT), amino acid transport protein (AAT), peptide transport Protein (PTR) and the like. The absorption and transmembrane transport of amino acids generated by the degradation of macromolecules in the source organ are mainly completed by amino acid transporters. In higher plants, AAT is a transmembrane protein that transports amino acids from the extracellular to the intracellular compartment, and also plays an important role in the long-distance transport of amino acids to the growing "pool", phloem loading of amino acids in the "source" organs during nitrogen mobilization, amino acid import during seed development, pathogenic responses, and abiotic stress.

AAT genes are divided into two superfamilies: the APC (amino acid, polyamine and choline transport) superfamily and the AAAP (amino acid/auxin permease) superfamily. The AAAP superfamily is divided into six subfamilies: AAPs (amino acid permeases) family, LHTs (lysine and histidine transporters) family, ProTs (proline transporters) family, GATs (gamma-amino acid butyric acid, GABA) family, AUXs (auxin transporters) family and ANTs (aromatic and neutral amino acid transporters) family. Research shows that the regulation of the expression of the amino acid transport protein coding gene can regulate and control the growth and development of transgenic plants and yield-related traits. For example, the rice yield and plant dry weight of T-DNA insertion mutants of OsAAT5, OsAAT7, OsAAT24, OsAAT49 and OsAAT60 are reduced, which proves that AAT plays an important role in nitrogen accumulation and carbon-nitrogen distribution of rice. Overexpression of AtLHTl in Arabidopsis thaliana can enhance the absorption function of transgenic plants to various amino acids, and increase the biomass accumulation of the transgenic plants (Hinier et al, 2006); the AAPl gene expressing kidney beans in peas significantly increases the total nitrogen and protein content in transgenic pea seeds, increasing seed weight (Rol Ietschek et al, 2005); overexpression of S1CAT9 in tomato fruits can improve the amino acid composition in fruits (Snowden et al, 2015). Therefore, the regulation and control of the expression level of the amino acid transporter gene have important significance on the improvement of the yield and the quality of crops. However, no studies on the influence of amino acid transporters on soybean plant types have been reported.

Disclosure of Invention

In order to solve the above problems, the present invention provides the use of a GmAAP protein and a GmAAP gene in soybean breeding. The invention explores the influence of the amino acid transporter on the soybean plant type, and the GmAAP gene plays an extremely important role in soybean branching, and can be applied to plant type improvement so as to improve the soybean yield.

In order to achieve the purpose, the invention provides the following technical scheme:

the invention provides an application of GmAAP protein in soybean breeding, wherein the amino acid sequence of the GmAAP protein is shown as SEQ ID NO. 1.

The invention also provides the application of the GmAAP gene in soybean breeding, wherein the cDNA sequence of the GmAAP gene is shown as SEQ ID NO. 2.

The invention also provides application of regulating the expression quantity of the GmAAP gene in improving the plant type of the soybean.

The invention also provides application of the silenced GmAAP gene in increasing the branching number of soybeans.

The invention also provides application of the silenced GmAAP gene in increasing the pod number of a single soybean plant.

The invention also provides application of the silenced GmAAP gene in improving the yield of soybean.

The invention provides an application of GmAAP protein in soybean breeding, wherein the amino acid sequence of the GmAAP protein is shown as SEQ ID NO. 1. The invention can obviously increase the branching number of the soybean by silencing the expression of the GmAAP gene through a gene engineering technology, thereby improving the yield of the soybean. The embodiment shows that the application provided by the invention can effectively improve the branch number of the soybeans and the pod number of a single plant.

Drawings

FIG. 1 is a structural diagram of main functional elements of a gene editing vector, wherein 35S, atU6 and Ubi are promoters, respectively; bar is a marker gene, and gRNA is a target sequence; dpCas9 is a Cas9 gene;

FIG. 2 shows a recombinant plant expression vector DTS 6001-GmAAP;

FIG. 3 is a table of the entire plant of soybean wild type Jack (OE-CK) and soybean wild type variety Williams82(KO-CK), 3 lines of GmAAP gene overexpression plants (OE-1, OE-2 and OE-3) and 3 lines of GmAAP gene silencing plants (KO-1, KO-2 and KO-3);

FIG. 4 is a statistical chart of the number of individual branches of each plant;

FIG. 5 is a statistical chart of the number of pods per plant;

FIG. 6 shows the relative expression level of GmAAP gene of each plant.

Detailed Description

The invention provides an application of GmAAP protein in soybean breeding, wherein the amino acid sequence of the GmAAP protein is shown as SEQ ID NO: 1: > Glyma.04G209100.1487aa

MVEYASRTNLSYCRDYDIEEDSMDGMPLKSDPECYDDDGRLKRTGTIWTTSSHIITAVVGSGVLSLAWAIAQMGWIAGPAVMILFSIVTLYTSSFLADCYRTGDPIFGKRNYTFMDAVSTILGGYSVTFCGIVQYLNLFGSAIGYTIAASLSMKAIQRSHCIIQFSDGENQCHIPSIPYMIGFGAVQIFFSQIPDFHNMWWLSIVASVMSFTYSIIGLVLGVTKIAETGTFKGSLTGISIGTVTEAQKVWGVFQALGNIAFAYSYSFVLLEIQDTIKSPPSEVKTMKKAAKLSIAVTTTFYMLCGCVGYAAFGDSAPGNLLAGFGFHKLYWLIDIANAAIVIHLVGAYQVYAQPLFAFVEKEAAKRWPKIDKEFQISIPGLQSYNQNVFSLVWRTVFVIITTVISMLLPFFNDILGVIGALGFWPLTVYFPVEMYILQKRIPKWSMRWISLELLSVVCLIVTIAAGLGSMVGVLLDLQKYKPFSSDY, SEQ ID NO: 1. The method of soybean breeding preferably comprises reducing expression of a GmAAP protein in a plant. In the present invention, under the precondition that the activity of the GmAAP protein is not affected (i.e. not in the active center of the protein), the skilled person can carry out various substitutions, additions and/or deletions of one or more amino acids on the amino acid sequence shown in SEQ ID NO.1 to obtain an amino acid sequence with equivalent functions. Therefore, the GmAAP protein also comprises a protein with equivalent activity obtained by substituting, replacing and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID NO. 1.

The invention also provides an application of the GmAAP gene in soybean breeding, wherein the cDNA sequence of the GmAAP gene is shown as SEQ ID NO: 2: > Glyma.04G209100.1CDS1464bp

ATGGTAGAATATGCTTCGAGAACAAACCTTAGCTACTGTCGAGATTATGACATTGAGGAGGACTCCATGGATGGCATGCCTTTAAAAAGTGATCCTGAATGCTATGACGATGATGGCCGTCTTAAACGAACAGGGACCATTTGGACTACAAGCTCCCACATAATAACAGCTGTGGTAGGATCTGGGGTGCTCTCCTTAGCCTGGGCAATAGCTCAGATGGGTTGGATTGCTGGTCCTGCAGTGATGATCTTATTCAGCATAGTCACTTTGTATACTTCATCATTTCTAGCTGATTGTTATCGTACTGGTGACCCCATATTCGGGAAGAGAAATTATACTTTCATGGATGCAGTTAGCACCATTCTAGGCGGGTACAGTGTTACGTTCTGTGGGATAGTTCAGTACTTAAATCTTTTCGGAAGTGCGATAGGATACACAATTGCGGCTTCCCTTAGCATGAAGGCAATCCAAAGGTCTCACTGTATCATCCAATTCTCTGATGGAGAAAACCAATGTCATATTCCAAGTATCCCATACATGATCGGTTTTGGTGCAGTGCAAATTTTCTTTTCTCAAATTCCAGATTTTCATAACATGTGGTGGCTCTCAATAGTTGCTTCAGTCATGTCTTTCACCTATTCCATAATTGGTCTCGTTCTTGGAGTTACCAAAATTGCAGAAACGGGAACTTTCAAGGGTAGCCTCACTGGAATAAGCATTGGAACTGTGACAGAGGCCCAAAAAGTATGGGGTGTTTTCCAAGCTCTTGGTAACATAGCCTTCGCCTATTCATATTCTTTCGTTCTCCTTGAAATTCAGGATACCATCAAATCTCCACCATCTGAAGTAAAAACAATGAAGAAGGCTGCAAAATTAAGTATTGCAGTGACCACAACATTTTATATGCTTTGTGGCTGCGTAGGCTATGCTGCTTTTGGGGATTCAGCACCTGGGAACCTGCTTGCTGGATTTGGTTTCCATAAACTATATTGGCTTATAGATATTGCTAATGCTGCTATTGTAATTCACCTTGTGGGGGCATACCAAGTGTATGCTCAACCCCTCTTTGCATTTGTCGAGAAGGAGGCAGCAAAAAGATGGCCCAAAATTGACAAGGAATTCCAAATTTCAATTCCCGGTTTGCAATCCTACAATCAGAACGTATTTAGCCTAGTTTGGAGGACAGTGTTTGTGATCATAACCACTGTTATATCAATGTTGCTTCCATTCTTCAATGATATTTTGGGAGTGATTGGAGCATTGGGGTTTTGGCCTCTAACGGTGTACTTTCCTGTGGAGATGTATATCTTGCAAAAGAGGATCCCAAAATGGAGTATGAGATGGATTTCTCTGGAATTGCTGAGTGTGGTGTGCCTCATAGTAACAATTGCGGCTGGTCTTGGCTCAATGGTTGGTGTCTTGCTTGACCTCCAGAAATACAAACCATTCAGTTCAGATTATTAA, SEQ ID NO: 2. The soybean breeding method preferably comprises silencing the GmAAP gene to obtain a transgenic plant with the GmAAP gene silenced. In the present invention, in consideration of the degeneracy of codons and the preference of codons for different species, one skilled in the art can use codons suitable for the expression of a particular species as needed.

The invention also provides application of regulating the expression quantity of the GmAAP gene in improving the plant type of the soybean. The application provided by the invention can effectively improve the plant type of the soybean, so that the plant type of the soybean subjected to gene improvement is shorter and stronger, more branches are formed, and the number of pods per plant is increased compared with the plant type of the soybean not subjected to improvement.

In the present invention, in the application of the silencing GmAAP gene in increasing the number of soybean branches or increasing the number of soybean single pods, the specific implementation method of silencing the GmAAP gene preferably includes: the method is characterized in that an amino acid transporter gene GmAAP of soybean is taken as an object, a cDNA sequence of the GmAAP is cloned from a soybean wild type variety Williams82, a GmAAP gene silencing expression vector is constructed by a Cas9 gene editing technology, an agrobacterium EHA105 mediated genetic transformation method is adopted, the Cas9 editing expression vector is introduced into a plant of a normal soybean wild type variety Williams82 to obtain a GmAAP gene silencing transgenic plant, the branch number of the soybean of the GmAAP gene silencing transgenic plant can be increased by 50% compared with that of the soybean wild type variety Williams82, and the single plant pod number is increased by 57.5% compared with that of the soybean wild type variety Williams 82.

The invention also provides application of the silenced GmAAP gene in improving the yield of soybean. The application provided by the invention can effectively improve the branch number and the pod number of the soybean, thereby improving the yield of the soybean. The invention also constructs a GmAAP gene over-expression vector, and the over-expression vector is introduced into soybean wild type Jack to obtain a GmAAP gene over-expression transgenic plant, wherein the branch number of the GmAAP gene over-expression transgenic plant is not obviously changed compared with the soybean wild type Jack. The above results indicate that the branch number of soybean can be increased by reducing the expression of the GmAAP gene, thereby improving the pod number per plant and the yield of soybean.

The invention also defines the molecular action mechanism of plant yield increase and has great promotion effect on defining the molecular action mechanism of plant yield increase.

In order to further illustrate the present invention, the following detailed description of the application of the GmAAP protein and GmAAP gene provided by the present invention in soybean breeding is provided with reference to the drawings and examples, but they should not be construed as limiting the scope of the present invention.

Examples

Construction of GmAAP gene silencing plant

The target of the gene is designed by using online website (http:// CRISPR. hzau. edu. cn/CRISPR2/) according to the GmAAP gene information in soybean reference genome in a soybean genome database (https:// www.soybase.org).

The target design principle is as follows:

1) the knock-out site is in the Coding (CDS) region and is as far as possible in the front end of the protein or in the important function domain region;

2) trying to cover a higher proportion of transcripts;

3) no off-target or off-target in the intergenic region;

4) selecting targets with higher editing efficiency;

5) the sequences have a more balanced GC content and are less prone to secondary structure formation.

The target sequence is shown as SEQ ID NO. 3:

gRNA：5'-ATGTCATATTCCAAGTATCC-3'，SEQ ID NO:3。

the structure diagram of the main functional elements of the gene editing vector is shown in FIG. 1, wherein 35S, atU6 and Ubi are promoters respectively; bar is a marker gene, and gRNA is a target sequence; dpCas9 is Cas9 gene.

According to the instructions in a CRISPR/Cas9 rapid construction kit VK005-04 (purchased from Shangrid biosciences), a gRNA shown in SEQ ID NO:3 is inserted into a gene editing vector to construct a CRISPR/Cas9 vector containing a GmAAP target sequence. A genetic transformation method mediated by agrobacterium EHA105 is adopted to introduce a gene silencing vector into a normal soybean wild type variety Williams82 to obtain a transgenic seedling.

Editing a plant obtaining process:

transplanting all the obtained transgenic plantlets in a basket with soil, watering and fertilizing at regular intervals, planting the transgenic plantlets in a field when the transgenic plantlets are about 10cm high, and utilizing the fertilizer after the transgenic plantlets grow up

The Bar test strip (EnviroLogix inc., USA) detects Bar protein by the following method:

(1) taking leaves of the grown transgenic plantlets as samples, placing sample leaf tissues between a cover and a tube body of a disposable tissue extraction tube, quickly covering the cover to obtain circular leaf tissues, and placing the leaves at the bottom of the extraction tube by using a pestle;

(2) and (3) inserting the pestle into the tube, rotating the pestle to grind the crushing blade, and continuously pressing for 20-30 seconds. Adding 0.5mL of extraction buffer solution;

(3) repeating the grinding step to fully contact and mix the sample with the buffer solution, and removing the pestle;

(4) the reaction tube is kept upright, the test paper is inserted into the reaction tube, the sample liquid rises along the test paper, and the result is read after the reaction is carried out for 10 minutes;

extracting genome DNA, detecting Bar in the transgenic plant through PCR, wherein a detection primer pair is shown as SEQ ID NO. 4 and SEQ ID NO. 5:

F3：5'-TGCACCATCGTCAACCACTACAT-3'，SEQ ID NO:4；

R3：5'-AGAAACCCACGTCATGCCAGT-3'，SEQ ID NO:5。

and detecting that the transgenic plant is possible to be a positive plant if the 480bp fragment is obtained by amplification. Then, collecting and planting a single plant, sampling and extracting DAN, sequencing nearby a target sequence by using a sequencing primer, and determining that the sequencing double peaks or the change of insertion or deletion of a base sequence nearby the target sequence is a positive plant. The sequencing primer pair is shown as SEQ ID NO. 6 and SEQ ID NO. 7:

F4：5'-TACACAATTGCGGCTTCCCT-3'，SEQ ID NO:6；

R4：5'-TGCAGGGGTTATGTGCCAAT-3'，SEQ ID NO:7。

and identifying homozygous transgenic plants until T2 generation to obtain GmAAP gene editing plants, subsequently selecting three lines and respectively marking the three lines as KO-1, KO-2 and KO-3, and marking the wild soybean variety Williams82 plant which is not subjected to GmAAP gene editing as KO-CK.

Construction of GmAAP gene over-expression plant

Extracting RNA of a soybean wild type variety Jack, reversely transcribing the RNA into cDNA, and performing reverse transcription on the cDNA by using a primer pair shown as SEQ ID NO. 8 and SEQ ID NO. 9:

F5：5'-TTTGGAGAGAACACGTATGGCTGAGCTTCACTACCA AC-3'，SEQ ID NO:8；

R5：5'-TCGGGGAAATTCGGGGTTAATAATCTGAACTGAATG-3'，SEQ ID NO:9。

the GmAAP gene is positively introduced into a soybean expression vector pCAMBIA3300 through a seamless cloning technology to construct a recombinant plant expression vector DTS6001-GmAAP (as shown in figure 2, the recombinant plant expression vector DTS6001-GmAAP) vector which is provided with a selective marker gene 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in a T-DNA region, wherein the gene codes the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) which can block the interference of glyphosate on a biosynthesis pathway, so that the glyphosate is not killed. And introducing the overexpression vector into a normal soybean wild type variety Jack by adopting an agrobacterium EHA105 mediated genetic transformation method to obtain a transgenic seedling of the soybean wild type variety Jack.

Transplanting all the obtained transgenic seedlings of the soybean wild type variety Jack into a basket with soil, watering and fertilizing at regular intervals, planting the seedlings in a field when the seedlings are about 10cm long, extracting genome DNA to detect transgenic plants through PCR after the seedlings grow up, wherein the detection primer pairs are shown as SEQ ID NO. 10 and SEQ ID NO. 11:

F3：5'-GACGCACAATCCCACTATCC-3'，SEQ ID NO:10；

R3：5'-TTAATAATCTGAACTGAATGGT-3'，SEQ ID NO:11。

if 1656bp segment is amplified, the transgenic plant is a positive plant. And (3) harvesting and planting a single positive plant until a homozygous transgenic plant is identified in the T2 generation, namely obtaining a hyper-expression plant of the GmAAP gene, subsequently selecting three plants which are respectively marked as OE-1, OE-2 and OE-3, and marking the Jack plant of the soybean wild type variety which is not edited by the GmAAP gene as OE-CK.

Application example 1

The plants of 3 lines (OE-1, OE-2 and OE-3) of soybean wild type variety Jack (OE-CK) and GmAAP gene overexpression plants provided in the example and 3 lines (KO-1, KO-2 and KO-3) of soybean wild type variety Williams82(KO-CK) and GmAAP gene silencing plants are cultivated, the cultivation results are shown in figure 3, figure 4, figure 5, table 1 and table 2, FIG. 3 is a table diagram of the entire strains of soybean wild type Jack (OE-CK) and soybean wild type Williams82(KO-CK), 3 strains of GmAAP gene overexpression plants (OE-1, OE-2 and OE-3) and 3 strains of GmAAP gene silencing plants (KO-1, KO-2 and KO-3), FIG. 4 is a statistical chart of the number of branches per plant, and FIG. 5 is a statistical chart of the number of pods per plant.

TABLE 1 statistical results of the number of individual branches of each plant

Sub-group (number)	Average number of branches per plant
		OE-CK(1)	5.20
OE-1(2)	3.20
		OE-2(3)	3.60
OE-3(4)	3.80
		KO-CK(5)	6.20
KO-1(6)	9.00
		KO-2(7)	9.20
KO-3(8)	9.40

As can be seen from FIGS. 3, 4 and Table 1, the number of branches of the plants (KO-1, KO-2 and KO-3) edited by the GmAAP gene was significantly greater than that of the wild type variety of soybean, Williams 82. The branch number of the GmAAP gene over-expression plant is not reduced significantly compared with the wild type variety Jack of soybean. Therefore, the silencing GmAAP gene can effectively increase the branching number of soybean.

TABLE 2 statistics of the number of individual pods per plant

As can be seen from FIG. 5 and Table 2, the number of pods per plant of the plants edited by the GmAAP gene (KO-1, KO-2 and KO-3) was significantly greater than that of the wild type variety of soybean, Williams 82. Compared with the wild soybean variety Jack, the branch number of the GmAAP gene over-expression plant is not significantly different. Therefore, the silencing of the GmAAP gene can effectively increase the single pod number of the soybean.

Application example 2

Taking a leaf of a GmAAP gene editing plant and a leaf of a GmAAP gene over-expression plant, extracting RNA, carrying out reverse transcription on the RNA to obtain cDNA, and detecting the expression quantity of the GmAAP gene in the editing plant and the over-expression plant through real-time fluorescent quantitative PCR. The detection results are shown in fig. 6 and table 3, and fig. 6 shows the relative expression level of the GmAAP gene of each plant. The primer pair used for the real-time fluorescent quantitative PCR is shown as SEQ ID NO. 12 and SEQ ID NO. 13:

F5：5'-TGGCCTCTAACGGTGTACTT-3'，SEQ ID NO:12；

R5：5'-AGACACCAACCATTGAGCCA-3'，SEQ ID NO:13。

TABLE 3 relative expression levels of GmAAP gene of each plant

As is clear from FIG. 6 and Table 3, the expression level of the GmAAP gene was lower in the gene-edited plants (KO-1, KO-2 and KO-3) than in the wild type soybean variety Williams82 plant which had not been subjected to gene editing. The expression quantity of the GmAAP gene in the over-expression plant is far higher than that of a soybean wild variety Jack.

The results show that the GmAAP gene can improve the branch number and the single pod number of the soybeans by reducing the expression level, and finally influences the yield of the soybeans.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Sequence listing

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<210> 7

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

tgcaggggtt atgtgccaat 20

<210> 8

<211> 38

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

tttggagaga acacgtatgg ctgagcttca ctaccaac 38

<210> 9

<211> 36

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

tcggggaaat tcggggttaa taatctgaac tgaatg 36

<210> 10

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

gacgcacaat cccactatcc 20

<210> 11

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

ttaataatct gaactgaatg gt 22

<210> 12

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

tggcctctaa cggtgtactt 20

<210> 13

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

agacaccaac cattgagcca 20

Claims

The application of the GmAAP protein in soybean breeding is shown in SEQ ID NO. 1.
2, the application of the GmAAP gene in soybean breeding, wherein the cDNA sequence of the GmAAP gene is shown as SEQ ID NO. 2.
3. The application of regulating the expression level of the GmAAP gene in improving the plant type of soybean.
4. Application of a silent GmAAP gene in increasing the branching number of soybeans.
5. Application of silent GmAAP gene in increasing pod number of single soybean plant.
6. Application of a silent GmAAP gene in improving yield of soybean.