Mint McTTG2 gene as well as expression protein and application thereof

Abstract

The invention discloses a mint McTTG2 gene as well as an expression protein and application thereof, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the mint McTTG2 gene disclosed by the invention is shown as SEQ ID NO. 1, and the amino acid sequence of the expression protein of the mint McTTG2 gene is shown as SEQ ID NO. 2. The method comprises the following steps: constructing an expression vector of a mint McTTG2 gene, and transforming the expression vector into an arabidopsis thaliana mutant; and cultivating and screening to obtain a transgenic arabidopsis thaliana plant of which the procyanidine content is obviously increased. A transgenic experiment shows that after the McTTG2 gene is over-expressed in the mutant ttg2-7, the content of procyanidine is remarkably increased; compared with the mutant ttg2-7, the number of epidermal hairs on leaves of ttg2-7 / 35S:: McTTG2-1 and-2 strains is obviously increased, which is close to that of wild strains. Gene resources are provided for plant variety improvement, and cultivation of new mint varieties is facilitated.

Description

Technical Field

[0001] The invention belongs to the technical field of plant genetic engineering, and more specifically relates to a mint McTTG2 gene and an expression protein and application thereof. Background Art

[0002] Mint (Mentha haplocalyx Briq.) is a perennial herbaceous plant in the genus Mentha (Mentha L.) of the Lamiaceae family. It is also widely planted as a medicinal and edible plant and a spice crop. Mint is a world-famous spice crop and an important medicinal and edible plant. It is widely planted worldwide. The whole herb of mint can be used as medicine, containing chemical components such as volatile oils, flavonoids, terpenes, phenolic acids, quinones, and phenylpropanoids. Among them, mint essential oil is an important secondary metabolite synthesized by mint. Its main component is volatile monoterpenoid compounds, which have antibacterial, antibacterial and fresh-keeping effects. It is used in drug development and is also widely used in food and daily chemical industries as spices and flavors. It has important economic value. The domestic market demand for mint and mint essential oil is growing, but due to the single mint germplasm resources, variety degradation, and limited planting area in my country, the mint yield and oil yield are low, and the quality of essential oil is unstable. Not only is it unable to be self-sufficient, but it also needs to be imported to meet domestic demand. Therefore, carrying out genetic improvement and innovation of mint germplasm resources, cultivating high-quality and high-yield mint varieties, and improving essential oil production and quality will be beneficial to promoting the development of my country's mint industry.

[0003] Plant epidermal hairs are hair-like epidermal structures formed by the specific differentiation of epidermal cells on the surface of the above-ground parts of plants, such as leaves and stems. Epidermal hairs form an important physical barrier between the plant epidermis and the environment, which helps plants adapt to changes in the surrounding growth environment. They not only help plants defend against biological stresses such as herbivores or pathogens, but also help plants resist abiotic stresses such as high temperature, drought, high salt, and ultraviolet rays. Plant epidermal hairs are usually divided into non-glandular hairs and glandular hairs according to whether they have the function of synthesis and secretion. Many secondary metabolites from plants are synthesized, stored, or secreted from plant glandular hairs. Peppermint essential oil, as a very important plant secondary metabolite, is synthesized and stored in peppermint glandular hairs. Peppermint epidermal hairs usually have three types: non-glandular hairs, capitate glandular hairs, and peltate glandular hairs. Among them, capitate glandular hairs and peltate glandular hairs have the ability to synthesize essential oils, while peltate glandular hairs have a larger volume than capitate glandular hairs and are the main place for the synthesis and storage of peppermint essential oils.

[0004] Proanthocyanidins are a mixture of flavonoids, whose basic unit is flavan-3-ol compounds. They are widely found in plant stems, barks, flowers, peels and seeds. They have multiple biological functions and are widely used in the pharmaceutical, food, cosmetics and other markets. According to the degree of polymerization, anthocyanidins are usually divided into oligomeric proanthocyanidins and polymeric proanthocyanidins. The former usually contains 2-5 monomers and is soluble, while the latter contains more than 5 monomers and is insoluble. Plants can use proanthocyanidins to regulate seed dormancy and germination, and resist biological and non-biological stresses, such as UV protection, resistance to pathogenic microorganisms, and anti-predation. For humans, proanthocyanidins are natural antioxidants, nutritional enhancers, and DNA protectors. They can effectively scavenge oxidative free radicals, reduce blood cholesterol and blood sugar levels, protect the cardiovascular system and prevent cardiovascular diseases. In addition, proanthocyanidins also affect the taste of some fruits and processed foods, such as persimmons, grapes, strawberries, and wine.

[0005] WRKY transcription factors are a class of transcription factors unique to plants. They contain conserved WRKY domains and C2H2 or C2HC zinc finger domains in their N-terminal and C-terminal structures, respectively. They can specifically bind to promoter regions containing W-box (C / TTGACT / C), SURE (sugar-responsive cis-element, a sequence rich in AT) and other related elements, thereby regulating the expression of target genes. WRKY not only plays an important regulatory role in plant response to biotic and abiotic stresses, but also plays an important regulatory role in plant development and the synthesis of secondary metabolites.

[0006] At present, WRKY transcription factors have been identified in other medicinal plants, but there are few reports on WRKY transcription factors in mint, especially mint WRKY transcription factors that regulate epidermal hair development and proanthocyanidin synthesis. Summary of the invention

[0007] In view of the above problems existing in the prior art, the technical problem to be solved by the present invention is to provide a mint McTTG2 gene. Another technical problem to be solved by the present invention is to provide an expression protein of the mint McTTG2 gene. Another technical problem to be solved by the present invention is to provide an application of the mint McTTG2 gene for regulating the formation of plant epidermal hairs and the accumulation of proanthocyanidins.

[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0009] A mint McTTG2 gene, whose nucleotide sequence is shown in SEQ ID NO. 1.

[0010] The expressed protein of the mint McTTG2 gene has an amino acid sequence as shown in SEQ ID NO. 2.

[0011] A vector, recombinant bacteria or host cell containing the mint McTTG2 gene.

[0012] The application of mint McTTG2 gene in regulating the content of proanthocyanidins in plants includes:

[0013] 1) constructing an expression vector of the mint McTTG2 gene;

[0014] 2) Transforming the constructed mint McTTG2 gene expression vector into Arabidopsis mutants;

[0015] 3) Cultivate, screen and obtain transgenic Arabidopsis plants with significantly increased proanthocyanidin content.

[0016] The application of the mint McTTG2 gene in regulating the content of proanthocyanidins in plants can achieve the regulation of the content of proanthocyanidins in plants by regulating the expression of key genes for proanthocyanidin accumulation.

[0017] The key genes for proanthocyanidin accumulation are AtTT12 and AtTT13 genes.

[0018] The application of the mint McTTG2 gene in regulating the development of plant epidermal hairs can achieve the regulation of plant epidermal hair development by regulating the expression of regulatory factors related to the formation and branching of epidermal hairs.

[0019] The regulatory factors related to the formation and branching of epidermal hairs are AtGL3, AtEGL3, AtGL2, AtMYB23, AtBRK1, AtARP2 and AtWAVE2.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1) The expression vector of the mint McTTG2 gene constructed by the present invention was transformed into the Arabidopsis strain. Before the transgenic Arabidopsis plants were dyed, the seed coat color of the mutant ttg2-7 was lighter than that of WT, ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2; after DMACA dyeing, the seed coat coloring degree of WT, ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 strains was significantly higher than that of the mutant ttg2-7. After overexpressing the McTTG2 gene in the mutant ttg2-7, the proanthocyanidin content was significantly increased. The results showed that overexpression of the McTTG2 gene promoted the accumulation of proanthocyanidins.

[0022] 2) Compared with the mutant ttg2-7, AtTT12 and AtTT13 genes were significantly upregulated in the ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 lines, close to the expression in the wild type. The results showed that McTTG2 positively regulated the accumulation of proanthocyanidins in transgenic Arabidopsis seeds by upregulating the expression of AtTT12 and AtTT13 genes at the molecular level.

[0023] 3) The expression vector of the mint McTTG2 gene constructed by the present invention was transformed into Arabidopsis strains. Compared with the mutant ttg2-7, the ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 strains had significantly more hairs on the leaf epidermis, which was close to the wild type.

[0024] 4) Compared with the mutant ttg2-7, the expression levels of AtGL3, AtEGL3, AtGL2, AtMYB23, AtBRK1, AtARP2 and AtWAVE2 genes were increased in the ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 lines. The results showed that McTTG2 promoted the formation of epidermal hairs in transgenic Arabidopsis by upregulating the expression of the above genes at the molecular level. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is the electrophoresis detection diagram of McTTG2 gene CDS cloning (M is DNA Marker; 1 is the full-length sequence of McTTG2 gene CDS);

[0026] Figure 2 The diagram shows the construction and identification of the McTTG2 gene overexpression vector (M is a DNA marker, 1 is the recombinant plasmid pHellsgate8-McTTG2-GFP, and 2 is the recombinant plasmid pHellsgate8-McTTG2-GFP cut with XhoI);

[0027] Figure 3 This is a diagram of RNA level identification of McTTG2 transgenic Arabidopsis;

[0028] Figure 4 The diagrams show the development of leaf epidermal hairs of Arabidopsis wild type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 (A is the leaf surface of each line photographed by scanning electron microscope; B is the statistics of the number of leaf epidermal hairs of each line; C is the statistics of the proportion of the number of epidermal hairs in different branches of leaves of each line);

[0029] Figure 5The expression levels of genes related to epidermal hair development in Arabidopsis wild type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2;

[0030] Figure 6 The seed colors and DMACA staining of Arabidopsis wild type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2;

[0031] Figure 7 The graph shows the proanthocyanidin content in Arabidopsis wild type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2;

[0032] Figure 8 This is a graph showing the expression levels of genes related to proanthocyanidin synthesis in Arabidopsis wild type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. Unless otherwise specified in the following embodiments, the technical means used are conventional means well known to those skilled in the art.

[0034] The plant material used in this application, the mint variety "68-7", was collected in the nursery of the Institute of Botany, Chinese Academy of Sciences, Jiangsu Province, and provided by Liang Chengyuan's research group; the Arabidopsis thaliana ttg2-7 mutant (sparse epidermal hairs, abnormal morphology, and low proanthocyanidin content) was purchased from the Arabidopsis germplasm resource bank TAIR (The Arabidopsis Information Resource).

[0035] Example 1

[0036] 1. Total RNA extraction and cDNA acquisition

[0037] Total RNA of mint was extracted using a plant tissue total RNA extraction kit (Nanjing Novozyme Biotechnology Co., Ltd.). The extracted total RNA was used as a template for cDNA synthesis using the third-generation high-efficiency cDNA first-strand synthesis kit (genome-free) (Nanjing Novozyme Biotechnology Co., Ltd.).

[0038] 2. Amplification of the CDS sequence of the mint McTTG2 gene

[0039] According to the transcriptome sequence of the mint McTTG2 gene, the following primers were designed:

[0040] Forward: 5'-ATGATTGAAGTAGAGGAGAAAGAAAAAATC-3',

[0041] Reverse: 5'-TCAGTTCTTAGTACTCGTCTTGGATGCATC-3'.

[0042] The CDS sequence of McTTG2 gene was obtained by PCR. The electrophoresis diagram of PCR amplification is shown in Figure 1 shown.

[0043] The CDS sequence of the McTTG2 gene obtained by final sequencing is shown in SEQ ID NO.1, totaling 1248 bp; it encodes a total of 415 amino acids, as shown in the sequence listing SEQ ID NO.2.

[0044] Example 2

[0045] 1. Construction of mint McTTG2 gene overexpression vector

[0046] According to the CDS sequence of the McTTG2 gene and the multiple cloning site of the overexpression vector pHellsgate8-GFP (XhoI was selected as the restriction site), amplification primers were designed. The primer sequences are as follows:

[0047] Forward:

[0048] 5'-CATTTGGAGAGGACACGCTCGAGATGATTGAAGTAGAGGAG-3';

[0049] Reverse:

[0050] 5'-CTTGCTCACCATGAATTCCTCGAGGTTCTTAGTACTCGTCTTG-3'.

[0051] PCR amplified the target fragment of McTTG2 gene. Use XhoI to digest pHellsgate8-GFP vector at 37℃ for 3 h. Use Shanghai Biotech Gel Recovery Kit to recover PCR products and digestion products to obtain the target fragment and linearized vector fragment. Use ClonExpress II One Step Cloning Kit Homologous Recombination Kit to connect the above target fragment with pHellsgate8-GFP linear vector to construct 35S::McTTG2 overexpression vector. The connection system is as follows: 5× CE II buffer 2μL, linearized vector about 100~200ng, insert fragment about 50~100ng, Exnase II1μL, supplemented with ddH 2 O to 10μL, place at 37℃ for 30min, and then immediately transfer to ice. After the E. coli DH5α competent cells are thawed on ice, add the above recombinant product to the competent cells and mix well, incubate on ice for 30min, then heat shock at 42℃ for 45sec, cool on ice for 5min, add 500μL LB medium, place on a 37℃ shaker for 1h, spread the bacterial liquid on a plate containing 100mg / L spectinomycin, and incubate inverted in a 37℃ incubator for 12~16h. Use colony PCR to verify single colonies, pick positive colonies and inoculate them into LB liquid culture medium containing 100mg / L spectinomycin, shake and culture at 200rpm, 37℃ for 24h, extract plasmid enzyme digestion verification ( Figure 2 ), and then sequenced after verification. The correct sequence is the 35S::McTTG2 overexpression vector.

[0052] 2. Agrobacterium infection of Arabidopsis

[0053] The 35S::McTTG2 overexpression vector plasmid was electroporated to transform the competent Agrobacterium GV3101, and the positive strain was verified by colony PCR. The Arabidopsis thaliana ttg2-7 mutant plant was transformed by the floral dip method. First, the transformation medium was prepared. The components included 1 / 2MS culture medium, 0.01μg / mL BAP, 5% sucrose, 0.02% silwetL-77, and the pH was adjusted to 5.7 with KOH. The 35S::McTTG2 Agrobacterium positive strain was picked and placed in YEP liquid medium containing 100mg / L spectinomycin and 50mg / L rifampicin, and the culture was shaken at 28℃, 200rpm, for 16h. Take 0.5mL of the above bacterial solution and transfer it to 50mL of YEP liquid medium containing the above antibiotics, and incubate at 28℃, 200rpm to make the OD 600 The value reaches about 1.8~2. Collect the bacterial solution in a centrifuge tube, centrifuge at 5000rpm for 5 minutes, discard the supernatant, and resuspend the precipitate with transformation medium until the resuspended liquid OD is 600The Arabidopsis plants to be transformed were placed flat, and the flower buds were immersed in the transformation medium for 1 minute, followed by dark culture for 16-24 hours, and then normal culture was resumed.

[0054] 3. Screening and identification of positive plants

[0055] After the infected seeds were disinfected with chlorine, they were cultured on MS medium containing 50 mg / L kanamycin. The resistant positive seedlings were selected and transplanted into the soil. After the seeds were harvested, they were screened with MS medium containing 50 mg / L kanamycin. The transformed seedlings with a 3:1 separation ratio were selected and planted into the soil. After the seeds were harvested, they were screened with MS medium containing 50 mg / L kanamycin. The transformed seedlings with 100% kanamycin resistance were selected. The McTTG2 transgenic homozygous lines were named ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2. The RNA of the leaves of the homozygous transgenic positive lines was extracted, and its cDNA was obtained by reverse transcription. Quantitative primers were designed, and semi-quantitative verification was performed ( Figure 3 ). The quantitative primers are as follows:

[0056] Forward: 5'-GGACAGAAACAGGTTAAAGGAAGTG-3';

[0057] Reverse: 5'-TCTTCAAACGTTCTCTCCACATACT-3'.

[0058] The total volume of the PCR reaction system was 20 μL: 1 μL each of the upstream and downstream primers, 2×Hieff ® Robust PCR MasterMix 10 μL, cDNA 1μL, ddH 2 O 7 μL.

[0059] The PCR reaction was performed using a PCR instrument with the following program: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 30 s, with a total of 40 cycles of annealing and extension.

[0060] Example 3

[0061] 1. Effect of overexpression of McTTG2 gene on the formation of Arabidopsis epidermal hairs

[0062] Arabidopsis seeds (WT, wild-type WT, mutant ttg2-7, and McTTG2 heterologous complemented transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2) were disinfected with 75% ethanol and 2% sodium hypochlorite solution, planted on 1 / 2MS solid culture medium, and cultured in a light incubator for 7 days. After the seedlings were transferred to soil and grown for 2-3 weeks, the epidermal hairs of each line were observed using a cryo-scanning electron microscope.

[0063] The results are as follows Figure 4 As shown, compared with the mutant ttg2-7, the ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 lines had significantly more hairs on the leaf epidermis, close to the wild type.

[0064] qRT-PCR was used to detect the gene expression levels of regulatory factors related to the formation and branching of epidermal hairs (such as AtGL3, AtEGL3, AtGL2, AtMYB23, AtBRK1, AtARP2 and AtWAVE2) in each strain. Upregulation of these genes will promote the formation and normal branching of epidermal hairs, while the number and branching of epidermal hairs will decrease.

[0065] The results are as follows Figure 5 As shown in the figure, compared with the mutant ttg2-7, the expression levels of these genes in ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 were increased. The results showed that McTTG2 promoted the formation of epidermal hairs in transgenic Arabidopsis by upregulating the expression of AtGL3, AtEGL3, AtGL2, AtMYB23, AtBRK1, AtARP2 and AtWAVE2 at the molecular level.

[0066] 2. Effect of overexpression of McTTG2 gene on proanthocyanidin accumulation in Arabidopsis seeds

[0067] Arabidopsis seeds (WT, wild-type WT, mutant ttg2-7, and McTTG2 heterologous complement transgenic lines ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2) were collected, observed and photographed under a stereo microscope. Seeds of the above four lines were taken for DMACA staining. Prepare 0.1% (w / v) DMACA solution, weigh an appropriate amount of DMACA powder, and dissolve it in 0.12N HCl-MeOH solution. Take an appropriate amount of Arabidopsis seeds, immerse them in the DMACA solution, stain at room temperature for about 1.5 hours, wash with deionized water, observe under a stereo microscope, and take pictures. The sulfuric acid-vanillin method was further used to determine the content of proanthocyanidins in the seeds. Prepare 1mg / mL proanthocyanidin standard solution, dilute to 0, 0.025, 0.05, 0.125, 0.25, 0.5, 1mg / mL proanthocyanidin solution, and make a standard curve. Proanthocyanidins are extracted with hydrochloric acid-methanol (0.1% HCl, v / v). Weigh 1g of seeds, add the extract and grind thoroughly, centrifuge at 8000rpm for 15min, transfer the supernatant, continue to extract with the extract twice, centrifuge to take the supernatant, combine the supernatants, blow dry with nitrogen, and re-dissolve in methanol. Take 200μL of proanthocyanidin extract, add 500μL of 1% (w / v) vanillin-methanol and 500μL of 25% (v / v) H 2 SO 4- Methanol solution, 30℃ water bath, reaction for 15min, spectrophotometer to measure A 500 The proanthocyanidin content was calculated.

[0068] The results are as follows Figure 6 As shown in the figure, before staining, the seed coat color of the mutant ttg2-7 was lighter than that of WT, ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2; after DMACA staining, the seed coat coloring degree of WT, ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 was significantly higher than that of the mutant ttg2-7. These differences are due to the lower proanthocyanidin content of the seeds of the mutant ttg2-7 than that of the WT, ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2.

[0069] The results are as follows Figure 7As shown, the proanthocyanidin content of the wild type WT seed is about 0.292 mg / g, and the proanthocyanidin content of the mutant ttg2-7 seed is about 0.095 mg / g. After overexpressing the McTTG2 gene in the mutant ttg2-7, the proanthocyanidin content was significantly increased (ttg2-7 / 35S::McTTG2-1 is 0.176 mg / g, ttg2-7 / 35S::McTTG2-2 is 0.314 mg / g). The results show that overexpression of the McTTG2 gene promotes the accumulation of proanthocyanidins.

[0070] The expression levels of key genes (AtTT12 and AtTT13) regulating seed proanthocyanidin accumulation in each strain were detected by qRT-PCR. AtTT12 and AtTT13 genes are positive regulatory genes for proanthocyanidin synthesis in Arabidopsis seeds.

[0071] The results are as follows Figure 8 As shown in the figure, compared with the mutant ttg2-7, AtTT12 and AtTT13 genes were significantly upregulated in the ttg2-7 / 35S::McTTG2-1 and ttg2-7 / 35S::McTTG2-2 lines, close to the expression in the wild type. The results showed that McTTG2 positively regulated the accumulation of proanthocyanidins in transgenic Arabidopsis seeds by upregulating the expression of AtTT12 and AtTT13 genes at the molecular level.

[0072] The above description is only illustrative rather than restrictive of the present invention. Those skilled in the art will understand that many modifications, changes or equivalents may be made without departing from the spirit and scope defined by the appended claims, but all will fall within the scope of protection of the present invention.

Claims

1. A mint McTTG2 gene, the nucleotide sequence of which is shown in SEQ ID NO.

1.

2. The expressed protein of the mint McTTG2 gene according to claim 1, whose amino acid sequence is shown in SEQ ID NO.

2.

3. A vector, recombinant bacteria or host cell containing the mint McTTG2 gene according to claim 1.

4. Use of the mint McTTG2 gene according to claim 1 in regulating the content of proanthocyanidins in plants.

5. The use according to claim 4, characterized in that: include: 1) constructing an expression vector of the mint McTTG2 gene; 2) Transforming the constructed mint McTTG2 gene expression vector into Arabidopsis mutants; 3) Cultivate, screen and obtain transgenic Arabidopsis plants with significantly increased proanthocyanidin content.

6. The use according to claim 4, characterized in that: By regulating the expression of key genes for proanthocyanidin accumulation, the proanthocyanidin content in plants can be regulated.

7. The use according to claim 6, characterized in that: The key genes for proanthocyanidin accumulation are AtTT12 and AtTT13 genes.

8. Use of the mint McTTG2 gene according to claim 1 in regulating the development of plant epidermal hairs.

9. The use according to claim 8, characterized in that: By regulating the expression of regulatory factors related to the formation and branching of epidermal hairs, the development of plant epidermal hairs can be regulated.

10. The use according to claim 9, characterized in that: The regulatory factors related to the formation and branching of epidermal hairs are AtGL3, AtEGL3, AtGL2, AtMYB23, AtBRK1, AtARP2 and AtWAVE2.

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