Transcription factor McZFP4 regulating epidermal hair and root hair development and salt stress tolerance in mint and its application
By identifying and applying the transcription factor McZFP4 in mint, the development of epidermal fur and root hair was regulated, and the tolerance of mint under salt stress was solved, negative regulation of plants' salt stress tolerance was achieved, and physiological indicators under salt stress were reduced. Improves salt tolerance of plants.
Patent Information
- Application Number
- CN202410699494.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-05-31
- Publication Date
- 2024-12-27
- Estimated Expiration
- 2044-05-31
AI Technical Summary
The lack of ZFP transcription factors in mint that regulate epidermal and root hair development, especially in salt stress-tolerant conditions, affects plant growth and essential oil yield.
The transcription factor McZFP4 and its applications are identified and provided in mint. By regulating salt stress response gene expression, epidermal fur and root hair formation related gene expression, transgenic plants are constructed to reduce epidermal fur and root hair development, and thus improve the plant's salt stress tolerance ability.
By downregulating the expression of related genes, the number of epidermal fur and root hairs of transgenic plants is significantly reduced, and the osmotic regulation ability and oxidase activity under salt tolerance is reduced, which proves the negative regulatory effect of McZFP4 gene in plant salt tolerance and reduces salt stress resistance.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of plant genetic engineering, and more specifically relates to a transcription factor McZFP4 for regulating the development of epidermal hairs and root hairs and salt stress resistance in mint and an application thereof. Background Art
[0002] Mint (Mentha haplocalyx Briq.) is a perennial herb in the genus Mentha (Mentha L.) of the family Lamiaceae. It is also widely cultivated as a dual-purpose medicinal and edible plant and a spice crop. Mint and its essential oil are widely used in the fields of pharmaceuticals, food processing, and daily chemical industry, and play an important economic role. Due to the reduction in planting area and the single germplasm resources, my country's mint essential oil production accounts for less than 5% of the global market, and a large amount of essential oil needs to be imported to meet domestic demand. Therefore, it is beneficial to promote the development of my country's mint industry to carry out genetic improvement and innovation of mint germplasm resources, cultivate high-quality and high-yield mint varieties, and improve the yield and quality of essential oils.
[0003] Epidermal hairs are hair-like epidermal structures that are specifically differentiated from epidermal cells. They are divided into non-glandular hairs and glandular hairs according to whether they have the function of synthesis and secretion. Epidermal hairs are one of the effective means of defense for plants against changes in the growth environment. They not only play an important protective role in defending against biological stresses such as herbivores or pathogens, but also can resist non-biological stresses such as high temperature, drought, high salt, and ultraviolet rays. Among them, glandular hairs have the ability to synthesize, store and secrete many metabolites. The surface of mint leaves and stems is densely covered with three different types of epidermal hairs: non-glandular hairs, capitate glandular hairs and peltate glands, among which peltate glandular hairs are the most important organs for essential oil synthesis and storage.
[0004] Root hairs are highly specialized cell structures in the epidermal cells of plant roots and are an important part of the plant root system. Root hairs can expand the surface area of the root system in the soil, promote plant growth and absorption of nutrients and water, help to fix the root system, and mediate the interaction between the root system and microorganisms in the soil. When there is a lack of nutrients in the soil, the formation of root hairs on the epidermis of plant roots increases to increase the contact area between the roots and the soil, which is beneficial for plants to absorb nutrients in the soil. In addition, when nitrogen-fixing bacteria attach to the surface of the root hairs of nitrogen-fixing plants, the root hairs will change their morphology to help the nitrogen-fixing bacteria colonize in the plant body.
[0005] Salt stress is one of the important abiotic stresses that restricts plant growth and development as well as crop yield and quality. Salt stress inhibits plant water absorption, causing cell dehydration and leaf wilting; salt stress causes excessive accumulation of sodium ions, destroys water balance and osmotic regulation, and causes plant wilting; salt stress has an adverse effect on plant photosynthesis and nutrient synthesis capacity, interferes with plant energy metabolism, photosynthesis and respiration, and inhibits plant growth and development; salt stress also causes the accumulation of reactive oxygen free radicals, leading to damage to cell membranes, proteins and nucleic acids.
[0006] Transcription factors play an important role in regulating plant growth and development and coping with abiotic stresses. The ZFP transcription factor family is an important transcription factor family in plants, playing an important regulatory role in plant growth and development and coping with abiotic stresses. At present, ZFP transcription factors have been identified in other plants, but ZFP transcription factors in mint have not been reported, especially ZFP genes that simultaneously inhibit the development of epidermal hairs and root hairs and negatively regulate plant salt stress tolerance have not been reported. 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 transcription factor McZFP4 that regulates the development of epidermal hairs in mint. Another technical problem to be solved by the present invention is to provide a transcription factor McZFP4 that regulates the development of root hairs in mint. Another technical problem to be solved by the present invention is to provide a transcription factor McZFP4 that regulates the ability to tolerate salt stress in mint. Another technical problem to be solved by the present invention is to provide an expression protein of the mint transcription factor McZFP4. Another technical problem to be solved by the present invention is to provide an application of the mint transcription factor McZFP4 for screening new plant germplasm.
[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:
[0009] A mint transcription factor McZFP4, whose nucleotide sequence is shown as SEQ ID NO.1.
[0010] The encoded protein of the mint transcription factor McZFP4, whose amino acid sequence is shown in SEQ ID NO.2.
[0011] A vector, a recombinant bacterium or a host cell containing the mint transcription factor McZFP4.
[0012] Application of mint transcription factor McZFP4 in regulating plant salt stress tolerance.
[0013] The application achieves the regulation of plant salt stress tolerance by regulating the expression of key genes responding to salt stress.
[0014] Application of mint transcription factor McZFP4 in regulating plant epidermal hair development.
[0015] The application achieves the regulation of plant epidermal hair development by regulating the expression of epidermal hair formation regulatory factors.
[0016] Application of mint transcription factor McZFP4 in regulating plant root hair development.
[0017] The application achieves regulation of plant root hair development by regulating the expression of root hair formation regulatory factors.
[0018] Application of mint transcription factor McZFP4 in reducing plant osmotic regulation ability, oxidase activity and ROS scavenging ability.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] 1) The mint transcription factor McZFP4 disclosed in the present invention has a nucleotide sequence as shown in SEQ ID NO.1, and an amino acid sequence of the encoded protein as shown in SEQ ID NO.2. The present invention constructs an expression vector of the mint transcription factor McZFP4; transforms the constructed expression vector of the mint transcription factor McZFP4 into Arabidopsis; and cultivates, screens, and obtains transgenic Arabidopsis plants with reduced salt stress tolerance.
[0021] 2) Compared with the wild type, the McZFP4 transgenic lines constructed by the present invention have fewer epidermal hairs on the leaves of lines OE2 and OE3. McZFP4 can inhibit the formation of epidermal hairs in transgenic Arabidopsis by downregulating the expression of AtZFP1, AtZFP8, AtGIS, AtGIS2, AtGIS3, AtGL3, AtEGL3 and AtTTG2 at the molecular level.
[0022] 3) Compared with the wild type, the McZFP4 transgenic lines constructed by the present invention have reduced root hair numbers and lower average lengths in lines OE2 and OE3. AtTTG1, AtWER and AtZP1 were significantly upregulated in the McZFP4 transgenic Arabidopsis lines, and AtZFP5, AtTRY, AtCPC, AtRHD6, AtRSL4 and AtLRL3 were significantly downregulated in the McZFP4 transgenic Arabidopsis lines. This indicates that McZFP4 can inhibit the development of root hairs in transgenic Arabidopsis at the molecular level by upregulating the expression of AtTTG1, AtWER and AtZP1 and downregulating the expression of AtZFP5, AtTRY, AtCPC, AtRHD6, AtRSL4 and AtLRL3.
[0023] 4) Compared with the wild type, the McZFP4 transgenic lines constructed by the present invention showed no significant difference in the growth of seedlings of lines OE2 and OE3 and the wild type WT under control conditions, but under 150 mM NaCl treatment, the main root lengths of OE2 and OE3 were significantly shorter than those of WT, indicating that under 150 mM NaCl treatment conditions, the salt tolerance of the transgenic lines was significantly lower than that of the wild type.
[0024] 5) Compared with the wild type, the McZFP4 transgenic strain constructed by the present invention has no significant difference in proline and malondialdehyde content, superoxide dismutase and catalase activity under normal conditions; however, under the condition of 150mM NaCl treatment, the proline content and superoxide dismutase and catalase activities in the overexpression strains OE2 and OE3 are significantly lower than those in WT, while the malondialdehyde content is significantly higher than that in WT. These results indicate that overexpression of the McZFP4 gene leads to a decrease in the osmotic regulation ability, oxidase activity and ROS scavenging ability of the transgenic plants, indicating that the McZFP4 gene negatively regulates the salt tolerance of plants.
[0025] 6) Compared with the wild type, AtDREB1A, AtCOR15A, AtCOR15B, AtRD29A, AtRD29B, and AtRAB18 were significantly down-regulated in the McZFP4 transgenic Arabidopsis strain constructed by the present invention. This indicates that McZFP4 can reduce the expression level of key genes responding to salt stress at the molecular level, thereby reducing the salt stress resistance of the McZFP4 transgenic Arabidopsis strain. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] Figure 1 This is the electrophoresis detection diagram of McZFP4 gene CDS cloning (M is DNA Marker; 1 is the full-length sequence of McZFP4 gene CDS);
[0027] Figure 2 The diagram shows the construction and identification of the McZFP4 gene overexpression vector (M is a DNA marker; 1 is the recombinant plasmid pHellsgate8-McZFP4-GFP; 2 is the recombinant plasmid pHellsgate8-McZFP4-GFP cut with XhoI);
[0028] Figure 3 This is a diagram of RNA level identification of McZFP4 transgenic Arabidopsis;
[0029] Figure 4 The diagrams are for the development of leaf epidermal hairs of Arabidopsis wild-type WT and McZFP4 transgenic lines OE2 and OE3 (A is a surface image of the leaves of each line taken by scanning electron microscope; B is a statistical diagram of the number of leaf epidermal hairs of each line);
[0030] Figure 5 The expression levels of genes related to epidermal hair development in Arabidopsis wild type WT and McZFP4 transgenic lines OE2 and OE3;
[0031] Figure 6 Figures are of the root hair development of Arabidopsis wild-type WT and McZFP4 transgenic lines OE2 and OE3 (A is a picture of the root hairs of each line taken with a stereo microscope; B is a statistical chart of the number of root hairs of each line; C is a statistical chart of the length of root hairs of each line);
[0032] Figure 7 The expression levels of genes related to root hair development in Arabidopsis wild type WT and McZFP4 transgenic lines OE2 and OE3;
[0033] Figure 8 The diagrams are salt stress tolerance analysis of Arabidopsis wild type WT and McZFP4 transgenic lines OE2 and OE3 (A is a diagram of root growth of seedlings of each line grown for four days on 1 / 2MS medium containing or not containing 150 mM NaCl after 14 days of growth; B is a statistical diagram of root length of each line in Figure A);
[0034] Fig. 9 The diagram shows the physiological indexes of Arabidopsis wild type WT and McZFP4 transgenic lines OE2 and OE3 under salt treatment (A is proline content; B is malondialdehyde content; C is superoxide dismutase activity; D is catalase activity);
[0035] Fig.10 The expression levels of salt stress-related genes in Arabidopsis wild type WT and McZFP4 transgenic lines OE2 and OE3 under salt treatment. DETAILED DESCRIPTION
[0036] 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.
[0037] The mint and Arabidopsis thaliana used in this application were collected from the nursery of the Institute of Botany, Chinese Academy of Sciences, Jiangsu Province.
[0038] Example 1
[0039] 1. Total RNA extraction and cDNA acquisition
[0040] 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 and cDNA was synthesized using the third-generation high-efficiency cDNA first-strand synthesis kit (genome-free) (Nanjing Novozyme Biotechnology Co., Ltd.).
[0041] 2. Amplification of the CDS sequence of the mint transcription factor McZFP4
[0042] According to the transcriptome sequence of the mint transcription factor McZFP4, the following primers were designed:
[0043] Forward: 5'-ATGCCGAGTGAAGAAACTGGTGATC-3',
[0044] Reverse: 5'-CTAAAGTTTGAGATTTAAGTCGAGC-3'.
[0045] The CDS sequence of McZFP4 gene was obtained by PCR. The electrophoresis diagram of PCR amplification is shown in Figure 1 shown.
[0046] The CDS sequence of the McZFP4 gene obtained by final sequencing is shown in SEQ ID NO.1, which is 489 bp in total and encodes 162 amino acids in total, as shown in the sequence listing SEQ ID NO.2.
[0047] Example 2
[0048] 1. Construction of mint transcription factor McZFP4 overexpression vector
[0049] According to the CDS sequence of McZFP4 gene and the multiple cloning site of the overexpression vector pHellsgate8-GFP (XhoI was selected as the restriction site), the amplification primers were designed: Forward: 5'-TTTGGAGAGGACACGCTCGAGATGCCGAGTGAAGAAACTG-3'; Reverse: 5'-GCTCACCATGAATTCCTCGAGAAGTTTGAGATTTAAGTCG-3', and the target fragment of McZFP4 gene was amplified by PCR. The pHellsgate8-GFP vector was digested with XhoI at 37℃ for 3h. The PCR product and the digested product were recovered by gel recovery using Shanghai Biotech Gel Recovery Kit to obtain the target fragment and linearized vector fragment. The above target fragment was connected with the pHellsgate8-GFP linear vector using the ClonExpress IIOne Step Cloning Kit homologous recombination kit to construct the 35S::McZFP4 overexpression vector. The connection system is as follows: 5×CEII buffer 2μL, linearized vector about 100-200ng, insert fragment about 50-100ng, Exnase II 1μL, add ddH2O to 10μL, place at 37℃ for 30min and then transfer to ice immediately. 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 500μLLB medium, place on a 37℃ shaker for 1h, spread the bacterial solution on a plate containing 100mg / L spectinomycin, and invert and culture in a 37℃ incubator for 12-16h. Use colony PCR to verify the single colony, pick the positive colony and inoculate it into LB liquid culture medium containing 100mg / L spectinomycin, shake and culture at 200rpm, 37℃ for 24h, extract the plasmid enzyme cutting verification ( Figure 2 ), and then sequenced after verification. The correct sequence is the 35S::McZFP4 overexpression vector.
[0050] 2. Agrobacterium infection of Arabidopsis
[0051] The 35S::McZFP4 overexpression vector plasmid was electroporated to transform Agrobacterium GV3101 competent cells, and the positive strains were verified by colony PCR, and Arabidopsis 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::McZFP4 Agrobacterium positive strain was picked to 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 YEP liquid medium containing the above antibiotics, and shake it at 28℃, 200rpm to make the OD 600 The value reached about 1.8 to 2.0. 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 600 The Arabidopsis plants to be transformed were placed flat, and the flower buds were immersed in the transformation medium for 10 seconds, followed by dark culture for 16 to 24 hours, and then normal culture was resumed.
[0052] 3. Screening and identification of positive plants
[0053] 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. Transformed seedlings with 100% kanamycin resistance were selected. The McZFP4 transgenic homozygous lines were named OE2 and OE3. 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 qRT-PCR verification was performed ( Figure 3 ). The quantitative primers are as follows:
[0054] Forward: 5'-TGCCGAGTGAAGAAACTGGT-3';
[0055] Reverse: 5'-GCATTCTGGTGTCCTCCCAA-3'.
[0056] The total volume of the PCR reaction system was 20 μL: 1 μL of upstream and downstream primers, 10 μL of 2×GS AntiQ qPCR SYBR FastMix, 1 μL of cDNA, and 7 μL of ddH2O.
[0057] The PCR reaction was carried out using a fluorescent quantitative PCR instrument BIO-RAD CFX-Opus 96. The operating program was as follows: pre-denaturation at 95°C for 30 s, denaturation at 95°C for 10 s, annealing and extension at 60°C for a total of 30 s, denaturation, annealing and extension for a total of 40 cycles, and fluorescence signal collection at 72°C.
[0058] Example 3
[0059] 1. Effect of McZFP4 overexpression on Arabidopsis epidermal hair formation
[0060] Arabidopsis seeds (WT, OE2 and OE3) overexpressing the mint transcription factor McZFP4 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 3-4 weeks, the epidermal hairs of McZFP4 transgenic Arabidopsis lines and wild-type Arabidopsis were observed using a stereomicroscope.
[0061] The results are as follows Figure 4 As shown, McZFP4 transgenic lines OE2 and OE3 had reduced leaf epidermal hairs compared with the wild type.
[0062] qRT-PCR was used to detect the gene expression levels of positive regulatory factors related to epidermal hair formation (such as AtZFP1, AtZFP8, AtGIS, AtGIS2, AtGIS3, AtGL3, AtEGL3 and AtTTG2) in Arabidopsis wild-type and McZFP4 transgenic lines. Upregulation of these genes will promote the formation of epidermal hairs, while downregulation will reduce the formation of epidermal hairs.
[0063] The results are as follows Figure 5 As shown in the figure, compared with the wild type, these genes were significantly down-regulated in the McZFP4 transgenic Arabidopsis lines. This indicates that McZFP4 can inhibit the formation of epidermal hairs in transgenic Arabidopsis by down-regulating the expression of AtZFP1, AtZFP8, AtGIS, AtGIS2, AtGIS3, AtGL3, AtEGL3 and AtTTG2 at the molecular level.
[0064] 2. Effect of McZFP4 overexpression on root hair development in Arabidopsis
[0065] Arabidopsis seeds (WT, OE2 and OE3) overexpressing the mint transcription factor McZFP4 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 3-4 weeks, the root hairs of the McZFP4 transgenic Arabidopsis lines and wild-type Arabidopsis were observed using a stereomicroscope.
[0066] The results are as follows Figure 6 As shown, compared with the wild type, the number and average length of root hairs in McZFP4 transgenic lines OE2 and OE3 were reduced.
[0067] The gene expression levels of regulatory factors related to root hair development (such as AtTTG1, AtWER, AtZP1, AtZFP5, AtTRY, AtCPC, AtRHD6, AtRSL4 and AtLRL3) in Arabidopsis wild-type and McZFP4 transgenic lines were detected by qRT-PCR. AtTTG1, AtWER and AtZP1 negatively regulate root hair initiation, and upregulation of these genes inhibits root hair formation; AtZFP5, AtTRY and AtCPC positively regulate root hair initiation, and downregulation of these genes inhibits root hair formation; AtRHD6, AtRSL4 and AtLRL3 positively regulate root hair initiation and elongation, and upregulation of these genes inhibits root hair formation and elongation.
[0068] The results are as follows Figure 7 As shown in the figure, compared with the wild type, AtTTG1, AtWER and AtZP1 were significantly up-regulated in McZFP4 transgenic Arabidopsis lines, and AtZFP5, AtTRY, AtCPC, AtRHD6, AtRSL4 and AtLRL3 were significantly down-regulated in McZFP4 transgenic Arabidopsis lines. This indicates that McZFP4 can inhibit the root hair development of transgenic Arabidopsis at the molecular level by up-regulating the expression of AtTTG1, AtWER and AtZP1 and down-regulating the expression of AtZFP5, AtTRY, AtCPC, AtRHD6, AtRSL4 and AtLRL3.
[0069] 3. Effect of overexpression of McZFP4 gene on salt tolerance in Arabidopsis
[0070] Arabidopsis seeds (WT, OE2 and OE3) were disinfected with 75% ethanol and 2% sodium hypochlorite solution, planted on 1 / 2MS solid medium, and cultured in light medium for 4 days; then WT, OE2 and OE3 seedlings with the same growth and roughly the same root length were selected and moved to 1 / 2MS solid medium (control group) and 1 / 2MS solid medium supplemented with 150mM NaCl (treatment group), and cultured in light medium for 7 days; the root length was counted by taking pictures. The software used for root length statistics was Image J.
[0071] 1) The result is as follows Figure 8As shown, there was no significant difference in the growth of seedlings of McZFP4-overexpressing Arabidopsis lines OE2 and OE3 and wild-type WT under control conditions, but under 150 mM NaCl treatment, the main root lengths of OE2 and OE3 were significantly shorter than that of WT, indicating that under 150 mM NaCl treatment, the salt tolerance of the transgenic lines was significantly lower than that of the wild type.
[0072] 2) The proline content was determined using a proline (Pro) assay kit (colorimetric method) (Nanjing Jiancheng Bioengineering Institute); the malondialdehyde content was determined using a malondialdehyde (MDA) assay kit (TBA method) (Nanjing Jiancheng Bioengineering Institute); the superoxide dismutase activity was determined using (Nanjing Jiancheng Bioengineering Institute Co., Ltd.); the catalase activity was determined using a catalase (CAT) assay kit (Nanjing Jiancheng Bioengineering Institute Co., Ltd.).
[0073] The results are as follows Fig. 9 As shown in the figure, under normal conditions, there were no significant differences in the proline and malondialdehyde contents, superoxide dismutase and catalase activities between the McZFP4 overexpressing Arabidopsis lines OE2 and OE3 and the wild type WT; however, under 150 mM NaCl treatment conditions, the proline content and superoxide dismutase and catalase activities in the overexpressing lines OE2 and OE3 were significantly lower than those in the WT, while the malondialdehyde content was significantly higher than that in the WT. These results indicate that overexpression of the McZFP4 gene leads to a decrease in the osmotic regulation ability, oxidase activity and ROS scavenging ability of transgenic plants, indicating that the McZFP4 gene negatively regulates the salt tolerance of plants.
[0074] 3) qRT-PCR was used to detect the expression levels of key genes (such as AtDREB1A, AtCOR15A, AtCOR15B, AtRD29A, AtRD29B, and AtRAB18) that respond to salt stress in wild-type Arabidopsis and McZFP4 transgenic Arabidopsis lines. These genes play an important positive regulatory role in plant response to salt stress. Upregulation of these genes will increase salt tolerance, while downregulation will lead to decreased salt tolerance.
[0075] The results are as follows Fig.10 As shown in the figure, compared with the wild type, AtDREB1A, AtCOR15A, AtCOR15B, AtRD29A, AtRD29B, and AtRAB18 were significantly down-regulated in McZFP4 transgenic Arabidopsis lines. This indicates that McZFP4 can reduce the salt stress resistance of McZFP4 transgenic Arabidopsis lines by down-regulating the expression levels of key genes responding to salt stress at the molecular level.
[0076] 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 transcription factor McZFP4 , whose nucleotide sequence is shown in SEQ ID NO.
1.
2. The peppermint transcription factor according to claim 1 McZFP4 The amino acid sequence of the encoded protein is shown in SEQ ID NO.
2.
3. Containing the peppermint transcription factor according to claim 1 McZFP4 vector or recombinant bacteria.
4. Overexpression of the peppermint transcription factor of claim 1 McZFP4 Application in negatively regulating Arabidopsis thaliana's salt stress tolerance.
5. Overexpression of the peppermint transcription factor of claim 1 McZFP4 Application in negatively regulating epidermal hair development in Arabidopsis thaliana.
6. Overexpression of the peppermint transcription factor of claim 1 McZFP4 Application in negatively regulating root hair development in Arabidopsis thaliana.
7. Overexpression of the peppermint transcription factor of claim 1 McZFP4 The application of the invention is to reduce the osmotic regulation ability, superoxide dismutase and catalase activities and ROS scavenging ability of Arabidopsis thaliana under salt stress conditions.