Application of soybean GmGSTF8 gene in increasing anthocyanin content of plants

Abstract

The invention belongs to the technical field of gene engineering, and particularly relates to application of a soybean GmGSTF8 gene in increasing the anthocyanin content of plants. The invention discloses a gene GmGSTF8 capable of regulating and controlling the anthocyanin content of a plant, the nucleotide sequence of the gene GmGSTF8 is as shown in SEQ ID NO.1, and the amino acid sequence of encoded protein is as shown in SEQ ID NO.2. Functional verification is carried out on the soybean GmGSTF8 gene through a transgenic arabidopsis thaliana technology and a transgenic tobacco technology, it is found that the content of anthocyanin in plant cells can be increased after overexpression of the GmGSTF8 gene, and important significance is achieved for cultivation of new functional soybean varieties.

Description

Technical Field

[0001] The present invention belongs to the technical field of genetic engineering, and particularly relates to the application of soybean GmGSTF8 genes in increasing the anthocyanin content of plants. Background Art

[0002] Soybean is native to China and is known as one of the four major oil crops together with rapeseed, peanut, and sesame. It not only provides humans with highly valuable oil and high-quality protein but also is rich in special plant nutrients such as saponins, vitamin E, and flavonoid substances. Among the flavonoid substances contained in soybean, anthocyanins, proanthocyanidins, isoflavones, flavonols, etc. have been reported to have good nutritional value and pharmacological effects. For example, a large number of studies have shown that anthocyanins play an important role in many aspects such as anti-inflammatory, anti-diabetic, prevention of Alzheimer's disease, hypertension, and cardiovascular diseases, as well as prevention of mutations, cancers, and tumors.

[0003] Anthocyanins are usually composed of anthocyanidins (aglycones) and one or more sugar groups. As important flavonoid substances, anthocyanins are widely present in organs such as the flowers, fruits, and seeds of plants, making these organs show rich colors from red to purple, thus attracting pollinators and seed dispersers. With the improvement of people's health awareness and the change of consumption concepts, foods rich in anthocyanins such as pitaya, purple cabbage, purple sweet potato, black corn, and black beans are favored by consumers. Some germplasm resources rich in anthocyanins have also been created through molecular design breeding, such as "purple tomato" and "purple crystal rice", etc. However, although there are varieties with black seed coats in soybean, the cotyledons in the seeds cannot synthesize and accumulate anthocyanins. Therefore, the research on the synthesis and regulation of soybean anthocyanins has potential application value.

[0004] Research shows that glutathione S-transferase (GST) plays a key role in the transport process of flavonoid anthocyanins. Anthocyanins are synthesized by a multi-enzyme complex on the cytoplasmic side of the endoplasmic reticulum and then transported to the vacuole for storage; the vesicle transport, membrane transport protein, and glutathione S-transferase-mediated models are three mechanisms that are both interrelated and different for vacuolar storage of anthocyanins. Plants producing anthocyanins generally have anthocyanin-specific glutathione transferase proteins, such as maize BZ2, Arabidopsis thaliana TT19, and grape VvGST1. Among them, glutathione S-transferase TT19-like GSTs are responsible for transporting anthocyanins from the endoplasmic reticulum to the vacuole as carriers; BZ2 can bind anthocyanin-3-glucoside (Cy3G) to glutathione; Arabidopsis thaliana TT19 and grape VvGST1 can bind to anthocyanins. However, the characteristics and molecular functions of the soybean GmGSTFs gene family have not been fully clarified at present, which limits the further development of soybean breeding work. Summary of the Invention

[0005] To solve the above problems, the present invention provides an application of a soybean GmGSTF8 gene in increasing the anthocyanin content in plants. The present invention verifies the function of the soybean GmGSTF8 gene through transgenic Arabidopsis technology and transgenic tobacco technology, and finds that overexpression of the GmGSTF8 gene can increase the anthocyanin content in plant cells.

[0006] The first aspect of the present invention provides an application of a soybean GmGSTF8 gene in increasing the anthocyanin content in plants, and the nucleotide sequence of the soybean GmGSTF8 gene is shown in SEQ ID NO.1.

[0007] Further, the application is to overexpress the soybean GmGSTF8 gene in plants to increase the anthocyanin content of the plants.

[0008] The second aspect of the present invention provides an application of a protein encoded by the above-mentioned soybean GmGSTF8 gene in increasing the anthocyanin content in plants, and the amino acid sequence of the protein encoded by the soybean GmGSTF8 gene is shown in SEQ ID NO.2.

[0009] The third aspect of the present invention provides an application of a recombinant expression vector containing the above-mentioned soybean GmGSTF8 gene in increasing the anthocyanin content in plants.

[0010] The fourth aspect of the present invention provides an application of an engineered bacterium containing the above-mentioned soybean GmGSTF8 gene in increasing the anthocyanin content in plants.

[0011] Further, the plant is soybean, Arabidopsis or tobacco.

[0012] The fifth aspect of the present invention provides a method for cultivating a transgenic plant with a high anthocyanin content, comprising the following steps: Connect the above-mentioned soybean GmGSTF8 gene to an expression vector, infect plant tissues through the Agrobacterium-mediated transformation method, and obtain a transgenic plant with a high anthocyanin content after cultivation.

[0013] Further, the expression vector is pBI121 or pTF101.

[0014] Further, the expression vector is pBI121.

[0015] Further, the Agrobacterium is Agrobacterium GV3101 or Agrobacterium EHA105.

[0016] Furthermore, the Agrobacterium is Agrobacterium GV3101.

[0017] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention discloses for the first time a soybean having a nucleotide sequence as shown in SEQ ID NO.1 GmGSTF8 Genes are related to the synthesis of anthocyanin content in plants, and experiments have confirmed that overexpression of soybean GmGSTF8 After the gene is added, the anthocyanin content in plant cells can be increased. GmGSTF8 The study of gene function provides a new approach to molecular breeding for improving soybean appearance quality, enhancing soybean nutritional value, and improving soybean stress resistance. It also has potential application value in the development of functional foods and natural pigment extraction.

[0018] (2) Soybean GmGSTF8 The gene belongs to the soybean GmGSTs gene family. It has been reported that the soybean GmGSTs gene family has many key functions: in terms of detoxification function, it can catalyze the combination of glutathione with herbicides and environmental toxins to achieve detoxification metabolism; in the face of biotic stress, it can enhance resistance to pathogens and pests by regulating defense signal pathways and secondary metabolite synthesis; under abiotic stress, it maintains cell homeostasis by resisting oxidation and regulating osmotic pressure; it also participates in plant hormone metabolism and secondary metabolite transport. This invention reports for the first time the soybean GmGSTs gene family. GmGSTF1 , GmGSTF5 , GmGSTF7 , GmGSTF8 , GmGSTF10 and GmGSTF11 The genes are mainly involved in the synthesis of anthocyanins in different soybean tissues and stages, and GmGSTF8 The gene has the function of regulating the anthocyanin content of plants and is mainly responsible for the synthesis of anthocyanins in the early development of stems, flowers and seeds, enriching the molecular function research of the soybean GmGSTs gene family. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 This is an evolutionary analysis diagram of soybean GmGSTs gene family proteins and GST proteins related to anthocyanin synthesis in other species.

[0021] Figure 2Figure A shows the amino acid sequence alignment results of the proteins of the soybean GmGSTs gene family and the GST proteins related to anthocyanin synthesis in other species. Figure B is a continuation of Figure A. The proteins marked with green dots in the front are the proteins of the soybean GmGSTs gene family. The green background represents the GST-N-phi conserved domain, and the orange background represents the GST-N-phi conserved domain. The conserved amino acid sites are selected by the red box. The black dots indicate similar amino acid sites.

[0022] Figure 3 is GmGSTF8 The heatmap shows the analysis of the expression levels of genes in different tissues of soybean. From left to right in the horizontal row, the soybean tissue samples are root, stem 1, stem 2, leaf 1, leaf 2, leaf 3, flower 1, flower 2, flower 3, flower 4, flower 5, seed 1, seed 2, seed 3, seed 4, seed 5, pod 1, pod 2, pod 3, seed pod 1, seed pod 2, seed pod 3, cotyledon 1, cotyledon 2, leaf bud 1, leaf bud 2, leaf bud 3, and meristem. The genes marked with stars in the vertical column are the genes of the soybean GmGSTFs subfamily.

[0023] Figure 4 is the phenotype difference result of overexpressing soybean GmGSTF8 gene in Arabidopsis thaliana. From left to right in the upper row of pictures are the wild-type Arabidopsis thaliana plants (marked as WT), the tt19 mutant plants (marked as tt19), the transgenic plants overexpressing the GmGSTF8 gene in the tt19 mutant (marked as #2), and the transgenic plants overexpressing the GmGSTF8 gene in the tt19 mutant (marked as #5). From left to right in the middle row of pictures are the rosette leaves of the WT plants, the rosette leaves of the tt19 mutant, the rosette leaves of the #2 transgenic plants, and the rosette leaves of the #5 transgenic plants. From left to right in the lower row of pictures are the seeds harvested from the WT plants, the seeds harvested from the tt19 mutant, the seeds harvested from the #2 transgenic plants, and the seeds harvested from the #5 transgenic plants.

[0024] Figure 5 is the phenotype difference of overexpressing soybean GmGSTF8 gene in tobacco. The left leaf is the phenotype after co-injecting the tobacco leaf with the MYB gene and the GmGSTF8 gene. The right leaf is the phenotype after injecting the tobacco leaf with the MYB gene alone. Specific implementation manners

[0025] The following is a detailed description of the specific embodiments of the present invention. It should be understood that the protection scope of the present invention is not limited by the specific embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the protection scope of the present invention. In the experimental methods described in the embodiments of the present invention, unless otherwise specified, they are all conventional methods. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0026] Soybeans not only provide valuable oil and high-quality protein for humans, but also are rich in special phytochemicals such as saponins, vitamin E, and flavonoids. At present, the characteristics and molecular functions of the soybean GmGSTFs gene family have not been fully clarified, which limits the further development of soybean breeding work.

[0027] The present invention provides an application of a soybean GmGSTF8 gene in increasing the anthocyanin content in plants. First, the amino acid sequences of the soybean GmGSTFs gene family were analyzed with the GST amino acid sequences reported to control anthocyanin synthesis in other species, and it was found that GmGSTFs belongs to glutathione S-transferase that controls anthocyanin synthesis; then the relationship between the expression of GmGSTFs and the synthesis of anthocyanins in different tissues and developmental stages of soybeans and the structural genes related to anthocyanin synthesis was explored, and it was found that GmGSTF1 , GmGSTF5 , GmGSTF7 , GmGSTF8 , GmGSTF10 and GmGSTF11 genes are mainly involved in the synthesis of anthocyanins in different tissues and stages of soybeans, GmGSTF8 can participate in the synthesis of soybean anthocyanins, and is mainly responsible for the synthesis of anthocyanins in the early stages of soybean stems, flowers and seed development; finally, an GmGSTF8 overexpression vector and genetic engineering bacteria were constructed, and the functions of the soybean GmGSTF8 gene were verified by transgenic Arabidopsis technology and transgenic tobacco technology respectively. It was found that overexpression of the GmGSTF8 gene could increase the anthocyanin content in plant cells.

[0028] Example 1: Function exploration of the soybean GmGSTFs gene family 1. Phylogenetic tree analysis of GmGSTs To analyze the gene function of soybean GmGSTs, the amino acids of GmGSTs were aligned with those of GST proteins in other species, and the alignment results were converted by MEGA version 6 to construct a phylogenetic tree.

[0029] The results are as Figure 1As shown, GmGSTs can be divided into 7 subfamilies. Among them, GmGSTFs proteins cluster in the Phi branch with GST proteins that control anthocyanin synthesis in other species, and have a relatively high homology with AtTT19 that controls anthocyanin synthesis in Arabidopsis thaliana, suggesting that GmGSFTs may be glutathione transferases that potentially control anthocyanin synthesis in soybeans.

[0030] 2. Amino acid sequence homology alignment and conserved domain analysis of GmGSTFs The amino acid sequences of GmGSTF1, GmGSTF2, GmGSTF3, GmGSTF4, GmGSTF5, GmGSTF6, GmGSTF7, GmGSTF8, GmGSTF9, GmGSTF10, GmGSTF11, GmGSTF12, GmGSTF13, and GmGSTF14 in the soybean GmGSTFs gene family, as well as the sequences of AtTT19, PfGST1, PhAN9, CKmGST3, MrGST1, LcGST4, PpGST1, MdGSTF6, VviGST4, and CsGSTF1 of GST proteins that control anthocyanin synthesis in other species reported currently, were input into the online software Clustal Omega (Clustal Omega <Multiple Sequence Alignment<EMBL-EBI) for amino acid multiple sequence alignment. The alignment results are as Figure 2 shown.

[0031] As can be seen from Figure 2 it, the proteins encoded by GmGSTFs all contain a Thioredoxin-like superfamily conserved domain at the N-terminus, and this region contains a [P]x10[S] domain. In addition, generally, a relatively conserved motif, the GST-C-phi motif, is contained at the C-terminus of GST proteins that control anthocyanin synthesis. Corresponding conserved domains exist at the N-terminus and C-terminus of the GmGSFTs amino acid sequence, belonging to the same conserved motif as the sequences of AtTT19, PfGST1, PhAN9, MrGST1, LcGST4, PpGST1, MdGSTF6, VviGST4, and CsGST1 with reported functions, indicating that GmGSTFs belong to glutathione transferases that control anthocyanin synthesis.

[0032] 3. GmGSTFs control the synthesis of anthocyanin To more clearly study the relationship between the expression of GmGSTFs and the synthesis of anthocyanins in different tissues and different developmental stages of soybeans, as well as the relationship with the structural genes related to anthocyanin synthesis, the present invention collected soybean materials from different tissues and different developmental stages, analyzed the expression of GmGSTFs and the structural genes related to anthocyanin synthesis, and explored whether the GmGSTFs gene is specifically expressed in different tissues and different developmental stages of soybeans.

[0033] Figure 3 For GmGSTF8 Heatmap of the analysis of the expression levels of genes in different parts of soybeans. As shown in the figure, the structural genes related to anthocyanin synthesis, GmF3H, GmF3’H, GmDFR1, GmDFR2, GmF3’5’H and GmUGT78K1 are highly expressed in the early stages of soybean stem, flower and seed development, which is consistent with the accumulation pattern of anthocyanins, suggesting their participation in anthocyanin synthesis.

[0034] GmGSTF1 and GmGSTF5 and GmGSTF7 and GmGSTF8 and GmGSTF10 and GmGSTF11 The gene expression patterns are similar to those of the key node genes in anthocyanin synthesis and are consistent with the accumulation pattern of anthocyanins in different tissues and different developmental stages. Therefore GmGSTF1 and GmGSTF5 and GmGSTF7 and GmGSTF8 and GmGSTF10 and GmGSTF11 genes are mainly involved in the synthesis of anthocyanins in different tissues and different developmental stages of soybeans.

[0035] In summary, GmGSTFs can participate in the synthesis of anthocyanins in soybeans and are mainly responsible for the synthesis of anthocyanins in the early stages of soybean stem, flower and seed development. The subsequent verification of gene functions mainly focuses on GmGSTF8 carried out. GmGSTF8 The nucleotide sequence of the gene is shown in SEQ ID NO.1, and the amino acid sequence of the encoded protein is shown in SEQ ID NO.2.

[0036] SEQ ID NO.1: ATGGTGGTGAAGGTGTATGGTCCAGCAAGTGCAGCCTGTCCTCAAAGGGTGTTGGTTTGCCTCTTAGAGAAAGGAGTAGAGTTTGAACTTGTGCATGTTGATCTTGATCTAGGAGAGCATAAGAAACCTGAATTTCTTCTCAGACAGCCCTTTGGTCAAGTTCCAGCTGTGGAGGATGGTGATTTTAGGCTCTTCGAATCCAGGGCTATTATAAGGTACTATGCATCAAAATTTGCAGACCGTGGCCCTGACCTATTGGGTAAAACCTTAGAGGAGAGGGCCCTAGTGGAGCAGTGGCTTGAAGTAGAGGCACACAACTTCAACAATTTGTGCTTCAATATCATGTTTCAGCTTGTGATCCTACCAAAGATGGGTAAGCCTGGGGACCTGGCCTTGGCACACAAATGTGAGCAAGATCTAAAAAAGGTGCTTGATGTGTATGAAAGTAGGCTCTCTCAAAGCACATATCTTGCTGGAGATAACTTCACTTTGGCTGATCTTAGCCACCTTCCAGGGCTTGGACACCTCATTGAGGAAGCCAAATTGGGGCACTTGGTCACTGAGAGGAAGAATGTGAGTGCTTGGTGGGAGAAAATTTCAAGTAGGCCAGCTTGGAAGAAGTTAAAGGATTTGGTTCATTAA。

[0037] SEQ ID NO.2: MVVKVYGPASAACPQRVLVCLLEKGVEFELVHVDLDLGEHKKPEFLLRQPFGQVPAVEDGDFRLFESRAIIRYYASKFADRGPDLLGKTLEERALVEQWLEVEAHNFNNLCFNIMFQLVILPKMGKPGDLALAHKCEQDLKKVLDVYESRLSQSTYLAGDNFTLADLSHLPGLGHLIEEAKLGHLVTERKNVSAWWEKISSRPAWKKLKDLVH*。

[0038] Example 2: GmGSTF8 Construction of overexpression vector 1. Extraction of soybean RNA Extract soybean RNA according to the instructions of the all-in-one plant RNA extraction kit (purchased from CWBIO Co., Ltd., product number CW2114), measure the concentration and quality of the RNA solution using an ND-1000 spectrophotometer, and store it in a -80°C refrigerator.

[0039] 2. Synthesize cDNA Reverse transcribe soybean RNA into cDNA. Prepare the reaction solution in an RNase-free PCR tube according to Table 1, place it in a PCR instrument, react at 70°C for 5 min, and perform an ice bath after the reaction to obtain Reaction Solution 1.

[0040] Table 1 Reaction system for reverse transcription of mRNA into cDNA (the first step) After centrifuging Reaction Solution 1, add the reaction system in Table 2 to the PCR tube, mix well, place it in a PCR instrument, react at 42°C for 1 h, obtain the cDNA template after the reaction, and store it at -20°C.

[0041] Table 2 Reaction system for reverse transcription of mRNA into cDNA (the second step) 3. PCR amplification and recovery Perform PCR amplification according to the reaction system in Table 3 and the reaction conditions in Table 4; the nucleotide sequence of the upstream primer is shown in SEQ ID NO.3, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.4; SEQ ID NO.3: 5’-ACACGGGGGACCATATGATGGTGGTGAAGGTGTATGGTC-3’; SEQ ID NO.4: 5’-GATCGGGGAAATTCGAGCTCTTAATGAACCAAATCCTTTAAC-3’.

[0042] Table 3 Reaction system for PCR amplification of the target fragment Table 4 Reaction conditions for PCR amplification of the target fragment Note: - indicates that the corresponding step does not perform a cycle.

[0043] After the PCR reaction, electrophoresis separation was carried out using 0.8% agarose gel. After cutting out the target band, the DNA fragment was recovered according to the operation steps of the Quick Agarose Gel DNA Recovery Kit (purchased from CW Biotech Co., Ltd., product number CW2302). After the concentration and quality of the obtained DNA fragment solution were measured by an ND-1000 spectrophotometer, it was stored in a -20°C refrigerator.

[0044] 4. Soybean GmGSTF8 Ligation and transformation of the gene with the expression vector pBI121 The DNA fragment was ligated with the expression vector pBI121 according to the reaction system in Table 5. After the ligation reaction solution was placed at room temperature for 1 h, the recombinant expression vector pBI121- GmGSTF8 was obtained. The recombinant expression vector pBI121- GmGSTF8 was transferred into Escherichia coli DH5α cells and cultured overnight at 37°C. Single colonies were picked, verified by PCR, positive clones were selected for sequencing, and the bacterial strains and recombinant expression vectors were preserved.

[0045] Table 5 Ligation system Example 3: Arabidopsis thaliana genetic transformation (1) Agrobacterium transformation The recombinant expression vector pBI121- GmGSTF8 prepared in Example 2 was transferred into Agrobacterium tumefaciens GV3101 by heat shock method, and positive clones were screened and verified using kanamycin, gentamicin and rifampicin. The specific steps are as follows:

[0046] Agrobacterium tumefaciens GV3101 competent cells were taken out from the -80°C refrigerator and thawed on ice for 5 min; 500 ng of the pBI121- GmGSTF8 recombinant expression vector was added to every 50 μL of Agrobacterium competent cells. After pipetting and mixing evenly, it was placed in an ice bath for 30 min; transferred to liquid nitrogen for 5 min; placed in a 37°C water bath for 5 min, and then in an ice bath for 5 min; then 800 μL of YEB liquid medium was added respectively, and it was cultured on a shaker at 28°C with a rotation speed of 200 rpm for 3 h; centrifuged at 5000 rpm for 5 min using a centrifuge, and the supernatant was discarded. The collected bacterial cells were spread on a YEB plate medium containing kanamycin sulfate and rifampicin, and cultured at 28°C for 2 d. Positive clones were screened and verified, and the bacterial solution with correct PCR verification was stored at -80°C for later use.

[0047] (2) The bacterial solution obtained in step (1) was inoculated into 5 mL of LB liquid medium containing kanamycin and gentamicin and cultured overnight on a shaker at 30°C; the next day, the bacterial solution was transferred and amplified at 30°C until the OD of the bacterial solution 600When the value is 0.6, collect the bacterial cells by centrifugation at 4700 rpm for 15 min.

[0048] (3) Suspend the bacterial cells in Arabidopsis infiltration buffer. Transfer the bacterial cells obtained in step (2) to a 250 mL beaker, add 300 mL of Arabidopsis infiltration buffer to a position 1 cm below the beaker mouth, add 60 μL of surfactant Silwet-77, and stir the solution evenly.

[0049] (4) Select Arabidopsis tt19 mutants that have grown for 5 weeks. Remove the siliques of Arabidopsis and leave the flowers. Invert them into the Arabidopsis infiltration buffer and infiltrate for 3 min; after infiltration, place the Arabidopsis horizontally in a tray containing a small amount of distilled water, keep it in the dark at room temperature overnight, transfer it to the greenhouse for normal cultivation and observation the next day. Mark the transgenic plants overexpressing the soybean GmGSTF8 gene in the tt19 mutants as #2 and #5.

[0050] (5) After the transgenic Arabidopsis seeds mature, collect the seeds, record them as the T1 generation, air-dry them and store them in a refrigerator at 4°C.

[0051] As Figure 4 shown, in the tt19 mutants, due to the lack of anthocyanin accumulation, the base of the stem is cyan. Compared with the tt19 mutant plants, the bases of the stems of #2 and #5 transgenic plants are brown, indicating that overexpressing the soybean GmGSTF8 gene can repair the phenotype of the tt19 mutants.

[0052] Example 4: Tobacco genetic transformation S1. Suspend the bacterial cells obtained in step (2) of Example 3 in tobacco infiltration buffer, centrifuge again at 4700 rpm for 15 min to collect the bacterial cells to ensure that the residual antibiotics are completely removed. Then adjust the OD 600 value of the bacterial solution to 0.1 with tobacco infiltration buffer to obtain the infiltration bacterial solution for tobacco infiltration.

[0053] S2. Select Nicotiana benthamiana leaves that have grown for 5 weeks. Use a 1 mL syringe to suck the infiltration bacterial solution in step S1 and inject it into the tobacco from the lower epidermis of the leaves, and observe the phenotypic changes of the leaves.

[0054] As Figure 5 shown, after 5 days, phenotypic changes occurred in the leaves. Compared with the leaves of the control group ( MYB the gene was injected into the tobacco leaves alone), overexpression of GmGSTF8 deepened the red patches on the tobacco leaves, indicating that overexpression of GmGSTF8 can increase the anthocyanin content in plant cells.

[0055] Although the preferred embodiments of the present invention have been described, additional changes and modifications can be made to these embodiments by those skilled in the art once they learn of the basic creative concept.

[0056] Obviously, those skilled in the art can make various changes and deformations to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and deformations of the present invention fall within the scope of the claims of the present invention and its equivalent technologies, the present invention also intends to include these modifications and deformations.

Claims

1. Soybeans GmGSTF8 The application of the gene in increasing the anthocyanin content of plants is characterized in that: The soybean GmGSTF8 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

2. The use according to claim 1, characterized in that: The application is to overexpress soybean in plants GmGSTF8 Gene, increasing the anthocyanin content of the plant.

3. The soybean according to claim 1 GmGSTF8 The use of a gene-encoded protein in increasing the anthocyanin content of a plant is characterized in that: The soybean GmGSTF8 The amino acid sequence of the protein encoded by the gene is shown in SEQ ID NO.

2.

4. A soybean containing the soybean according to claim 1 GmGSTF8 Application of recombinant gene expression vector in increasing the anthocyanin content of plants.

5. A kind of soybean containing claim 1 GmGSTF8 The application of genetically engineered bacteria in increasing the anthocyanin content of plants.

6. The use according to claim 1, characterized in that: The plant is soybean, tobacco or Arabidopsis thaliana.

7. A method for cultivating transgenic plants with high anthocyanin content, characterized in that: The method comprises the following steps: GmGSTF8 The gene is connected to an expression vector, and after infecting plant tissues through an Agrobacterium-mediated transformation method, the transgenic plant with high anthocyanin content is obtained.

8. The method according to claim 7, characterized in that The expression vector is pBI121 or pTF101.

9. The method according to claim 7, characterized in that: The Agrobacterium is Agrobacterium GV3101 or Agrobacterium EHA105.

Images