WRKY transcription factor gene StWRKY61 and application thereof

Abstract

The application discloses a WRKY transcription factor gene StWRKY61 and application thereof, and belongs to the technical field of plant genetic engineering. The nucleotide sequence is shown as SEQ ID NO. 1. The application proves that overexpression of StWRKY61 can significantly improve the survival rate and biomass accumulation of plants under drought conditions, and can enhance the osmotic regulation ability and antioxidant defense system, thereby effectively relieving the damage caused by drought stress. The gene provides a new gene resource and technical approach for drought-resistant breeding of potatoes and other crops.

Description

Technical Field

[0001] This invention relates to the field of plant genetic engineering technology, and in particular to the WRKY transcription factor gene StWRKY61 and its applications. Background Technology

[0002] With global climate change, drought has become one of the major abiotic stress factors limiting crop yields. As a vital global food crop, improving the drought resistance of potatoes is of significant economic and social importance. Currently, methods to enhance plant drought resistance mainly include transgenic technology, breeding improvement, and cultivation management measures. Regarding transcription factors regulating plant drought resistance, studies have shown that certain transcription factors, such as SlWRKY80 and StMYB96, participate in plant drought resistance regulation. However, the specific function and regulatory mechanism of StWRKY61, a member of the WRKY family in potatoes, in drought stress response remain unclear. Existing technologies lack functional verification and application schemes for the potato transcription factor StWRKY61 in drought resistance, resulting in the untapped potential of this gene in drought-resistant breeding. Summary of the Invention

[0003] The purpose of this invention is to provide the WRKY transcription factor gene StWRKY61 and its applications to address the problems existing in the prior art. By constructing an overexpression system of StWRKY61 and introducing it into plants, the physiological tolerance and survival ability of plants under drought stress conditions are enhanced, providing new gene resources and technical support for drought-resistant crop breeding.

[0004] To achieve the above objectives, the present invention provides the following solution:

[0005] One of the technical solutions of the present invention is a WRKY61 gene that improves the drought resistance of potatoes, the nucleotide sequence of which is shown in SEQ ID NO.1.

[0006] The second technical solution of the present invention is a WRKY61 protein that improves the drought resistance of potatoes, the nucleotide sequence of which is shown in SEQ ID NO.2.

[0007] The third technical solution of the present invention includes a recombinant vector, expression cassette, transgenic cell line or recombinant bacteria containing the WRKY61 gene.

[0008] The fourth technical solution of the present invention is the application of the WRKY61 gene, the WRKY61 protein, or the recombinant vector, expression cassette, transgenic cell line, or recombinant bacteria in improving the drought resistance of potatoes by overexpressing the WRKY61 gene or increasing the level of the WRKY61 protein to improve the drought resistance of potatoes.

[0009] The fifth technical solution of the present invention is a method for improving the drought resistance of potatoes by overexpressing the WRKY61 gene or increasing the level of WRKY61 protein to improve the drought resistance of potatoes.

[0010] The sixth technical solution of the present invention is the application of the WRKY61 gene, the WRKY61 protein, or the recombinant vector, expression cassette, transgenic cell line, or recombinant bacteria in the cultivation of new potato varieties with strong drought resistance.

[0011] The seventh technical solution of the present invention is a method for cultivating a new potato variety with strong drought resistance, comprising the following steps: introducing the WRKY61 gene into the target plant, thereby obtaining the potato variety.

[0012] Based on the above technical solution, the present invention has the following technical effects:

[0013] This invention demonstrates that overexpression of StWRKY61 significantly improves plant survival rate and biomass accumulation under drought conditions, enhances osmotic regulation and antioxidant defense systems, thereby effectively alleviating damage caused by drought stress. This gene provides a new genetic resource and technical approach for drought-resistant breeding of potatoes and other crops. Attached Figure Description

[0014] Figure 1 This is a clone of the StWRKY61 gene. Note: M represents Marker, and 1 represents the target gene.

[0015] Figure 2 The double enzyme digestion verification of the potato StWRKY61 overexpression vector is shown. M represents Marker, 1 represents the undigested plasmid, and 2 represents the plasmid containing the target gene StWRKY61.

[0016] Figure 3 In the diagram, A represents the genetic transformation of the potato StWRKY61 overexpression vector, a represents callus tissue, b represents callus tissue differentiating into shoots, and c represents rooting screening of transformed plants; B represents the verification of transgenic plants, 1, 2, and 3 represent overexpression lines OE-1, OE-2, and OE-3; + represents positive control; - represents negative control; and M represents Marker.

[0017] Figure 4 The expression level of StWRKY61 in transgenic plants.

[0018] Figure 5 This study aimed to determine the phenotype, detached leaf water loss rate, and relative water content of transgenic potatoes under drought stress. In the figures, A represents the phenotype of transgenic potatoes after natural drought stress; B represents relative water content (RWC); and C represents the leaf water loss rate. Each column represents the mean + SD (n=3). Different lowercase letters indicate significant differences within groups (P < 0.05).

[0019] Figure 6 This study measured the phenotype, plant height, root length, and fresh weight of transgenic potatoes under simulated drought stress using PEG6000. In this study, A represents the phenotype of transgenic potatoes after natural drought stress; B represents plant height, root length, and fresh weight; each column represents the mean plus SD (n=3); different lowercase letters indicate significant differences within groups (P<0.05). Detailed Implementation

[0020] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.

[0021] This invention provides a WRKY61 gene that enhances the drought resistance of potatoes, the nucleotide sequence of which is shown in SEQ ID NO.1.

[0022] This invention also provides a WRKY61 protein that improves the drought resistance of potatoes, the nucleotide sequence of which is shown in SEQ ID NO.2.

[0023] This invention also provides the application of the WRKY61 gene, the WRKY61 protein, or the recombinant vector, expression cassette, transgenic cell line, or recombinant bacteria in improving the drought resistance of potatoes, by overexpressing the WRKY61 gene or increasing the level of the WRKY61 protein to improve the drought resistance of potatoes.

[0024] This invention also provides a method for improving the drought resistance of potatoes by overexpressing the WRKY61 gene or increasing the level of WRKY61 protein.

[0025] The present invention also provides the application of the WRKY61 gene, the WRKY61 protein, or the recombinant vector, expression cassette, transgenic cell line, or recombinant bacteria in the cultivation of new potato varieties with strong drought resistance.

[0026] This invention also provides a method for cultivating a new potato variety with strong drought resistance, comprising the following steps: introducing the WRKY61 gene into the target plant, thereby obtaining the potato variety.

[0027] In some specific implementations, the WRKY61 gene is introduced into the target plant via the recombinant vector, expression cassette, transgenic cell line, or recombinant bacteria.

[0028] Example 1

[0029] This invention clones the potato WRKY61 gene, constructs its overexpression vector, and transforms it into a model plant (potato itself) to verify its physiological and molecular functions under drought stress. The specific steps are as follows:

[0030] 1. Gene cloning: Total RNA was extracted from the potato variety "Atlantic", and cDNA was synthesized by reverse transcription. The complete coding sequence (CDS) of the StWRKY61 gene was obtained by amplification using specific primers.

[0031] 2. Vector construction: The purified StWRKY61 CDS was inserted into the multiple cloning site of the plant overexpression vector pC2300S-GFP by enzyme digestion and ligation to construct the recombinant plasmid pC2300S-StWRKY61;

[0032] 3. Genetic transformation: Potato microtubules (wild type) were transformed using Agrobacterium-mediated transformation. The constructed overexpression vector was introduced into potato microtubule explants, and transgenic plants were obtained through screening and regeneration.

[0033] 4. Analysis of gene expression levels in transgenic plants StWRKY61.

[0034] 5. Drought resistance assessment: Under drought stress, the phenotype, survival rate, relative leaf water content, water loss rate of detached leaves, and antioxidant enzyme (SOD, POD, CAT) activities of transgenic plants and wild-type plants were compared.

[0035] Specific experimental steps:

[0036] (a) Material preparation

[0037] 1. Plant materials: tissue culture seedlings and miniature tubers of the potato variety "Atlantic".

[0038] Wild-type (WT) and transgenic overexpression lines (OE-1, OE-2, OE-3).

[0039] 2. Strains and vectors: Escherichia coli competent cells DH5α, Agrobacterium strain GV3101.

[0040] Plant overexpression vector pC2300S-GFP (containing Kanamycin resistance).

[0041] 3. Reagents and enzymes: RNA extraction kit, reverse transcription kit, high-fidelity DNA polymerase; restriction endonucleases KpnⅠ and BamhⅠ, T4 DNA ligase or homologous recombinase; PCR purification kit, gel extraction kit, plasmid extraction kit; plant hormones, antibiotics (Kanamycin, Hygromycin, Cefotaxime); PEG6000, MS medium, LB medium.

[0042] 4. Instruments and equipment: PCR instrument, electrophoresis system, gel imaging system; clean bench, constant temperature shaker, centrifuge; light incubator, drying oven, analytical balance; real-time quantitative PCR instrument (qRT-PCR).

[0043] (II) Experimental Methods

[0044] 1. RNA extraction and cDNA synthesis and StWRKY61 gene cloning

[0045] Obtaining the target gene (StWRKY61CDS):

[0046] (1) Take young potato leaves, grind them in liquid nitrogen, and extract total RNA using an RNA extraction kit (when using the kit, be careful to prevent RNase contamination). Detect the integrity and concentration of RNA by agarose gel electrophoresis. Use a reverse transcription kit to reverse transcribe the RNA into cDNA first strand.

[0047] (2) Using the synthesized cDNA as a template, PCR amplification was performed using the designed primers (F: GCATCACCAAAGAGGCCAAG, R: TGCAGCTGTGAAAGTAGGGC, reference gene version number XM_006361470.1) and high-fidelity DNA polymerase.

[0048] PCR program: Set the number of cycles for pre-denaturation, denaturation, annealing, and extension according to the Tm value of the primers and the requirements of the polymerase.

[0049] Product validation: The size of the PCR product (786 bp) was verified by 1% agarose gel electrophoresis, and the target band was recovered by gel extraction (using a gel recovery kit).

[0050] 2. Construction of overexpression vectors

[0051] (1) Select a plant overexpression vector (pC2300S-GFP) suitable for Agrobacterium-mediated transformation or transient expression.

[0052] (2) Vector linearization: The pC2300S-GFP circular plasmid was linearized by double digestion of the vector with restriction endonucleases (KpnⅠ and BamhⅠ) at the multiple cloning site.

[0053] (3) Perform agarose gel electrophoresis on the enzyme digestion products, cut out the linearized vector band of the correct size, purify it using a gel recovery kit, and determine the concentration.

[0054] (4) Ligation of the target gene and the vector: The purified linearized vector and the purified target gene were mixed at a ratio of vector:insertion fragment = 1:3. Homologous recombinase was added and the mixture was reacted at 37°C for 50 minutes.

[0055] (5) Transform the recombinant vector into competent E. coli cells: Add 10 μL of the ligation product to competent E. coli cells (DH5α) and incubate on ice for 30 minutes. Heat shock at 42°C for 45 seconds, then immediately incubate on ice for 2 minutes. Add antibiotic-free LB liquid medium and incubate at 37°C with shaking for 60 minutes. Centrifuge the incubated culture and spread a portion of the cells onto LB solid plates containing Kan resistance. Incubate upside down at 37°C for 12-16 hours.

[0056] (6) Select a single clone and resuspend it in LB liquid medium containing Kan resistance and culture with shaking for 6 hours. Perform PCR amplification of the bacterial culture using universal primers for the vector (NPTⅡ-F, NPTⅡ-R). Detect the presence of bands of the expected size by agarose gel electrophoresis to preliminarily screen for positive clones.

[0057] (7) Send the striped bacterial culture to the sequencing company for sequencing.

[0058] (8) The strain that was verified to be completely correct by sequencing was the engineered strain that successfully constructed the StWRKY61 overexpression vector. Finally, plasmid DNA was extracted using a plasmid miniprep kit to obtain the StWRKY61 overexpression vector.

[0059] 3. Genetic transformation

[0060] (1) Agrobacterium transformation: The recombinant plasmid was introduced into Agrobacterium GV3101.

[0061] (2) Prepare miniature potato slices, inoculate them onto a co-culture medium, and culture them in the dark for 2 days.

[0062] (3) Pick a single Agrobacterium clone containing the recombinant plasmid from a fresh plate and inoculate it into LB liquid medium containing antibiotics. Incubate at 28°C and 240 rpm with shaking until the logarithmic growth phase (OD200). 600 (≈ 0.5), collect the bacterial cells by centrifugation, and resuspend them in an equal volume of infection medium (MS liquid medium).

[0063] (4) Soak the miniature potato slices in the prepared Agrobacterium tumefaciens solution and gently shake for 7 minutes. After infection, use sterile filter paper to absorb the excess bacterial solution on the surface of the explant.

[0064] (5) Transfer the infected miniature potato slices back to the co-culture medium and co-culture at 20-25°C for 2 days in the dark.

[0065] (6) After co-culture, it needs to be transferred to differentiation medium. When the resistant shoots grow to 1-2 cm in height, they are cut off and transferred to root sieve medium containing antibiotics (50 μL cephalosporin and 20 μL hygromycin) to allow them to root and obtain resistant seedlings.

[0066] (7) Identification of transgenic plants: Samples were taken, ground in liquid nitrogen, and genomic DNA was extracted from the resistant seedlings. PCR amplification was performed using universal primers (NPTⅡ-F, NPTⅡ-R) for the StWRKY61 gene. Plants with a 676bp band detected on gel electrophoresis were PCR-positive transgenic plants.

[0067] 4. Expression level analysis

[0068] (1) Total RNA was extracted from wild-type (WT) and transgenic lines (OE-1, OE-2, OE-3) and reverse transcribed to synthesize cDNA.

[0069] (2) Amplification reaction was performed on a real-time quantitative PCR instrument using StWRKY61 specific primers and internal reference gene EF1α primers.

[0070] (3) The relative expression level of the StWRKY61 gene in each line was calculated using the 2^–ΔΔCt method and standardized using WT as the benchmark.

[0071] 5. Identification of drought resistance phenotypes

[0072] (1) Natural drought treatment: transplant the tissue culture seedlings into the soil, water normally for 4 weeks, then stop watering and continue to be dry for 7 days, and take pictures to record the phenotype.

[0073] (2) PEG6000 simulated drought: The tubers of wild-type and overexpression plants were inoculated into 2% MS liquid medium, and three replicates were set up for the control group and the experimental group respectively. The control group grew normally for four weeks, the experimental group grew normally for one week, and was subjected to PEG6000 drought stress for three weeks. Then the phenotypes of the control group and the experimental group were photographed.

[0074] (3) Measurement of physiological indicators:

[0075] Root length and fresh weight of stems and leaves: The fresh weight of stems and leaves of plants in the control and experimental groups treated with PEG6000 drought was weighed, and the root length was measured. After recording the data, a bar chart was drawn using GraphPad Prism 8 software.

[0076] Relative Moisture Content (RWC): Leaf fragments of uniform size were quickly cut with scissors, with three technical replicates for each sample. After collection, the samples were immediately weighed on a balance, and the weight was recorded as FW: fresh weight. Sufficient distilled water was added to the container holding the weighed fresh leaves, ensuring all leaves were completely submerged. The container was placed at 4°C in the dark for 24 hours. After 24 hours of soaking, the leaves were gently removed with tweezers. All excess moisture was carefully blotted from the leaf surface with filter paper or paper towels, and the leaves were immediately weighed, recorded as saturated fresh weight TW. The leaves, after being weighed for saturated fresh weight, were placed in an oven preheated to 105°C and blanched for 30 minutes. Then, the temperature was lowered to 80°C, and drying continued for 48 hours until the sample reached constant weight. The sample was weighed again and recorded as dry weight DW. The formula is as follows: RWC (%) = [(FW - DW) / (TW - DW)] × 100%. After recording the data, use the software GraphPad Prism 8 to draw a bar chart.

[0077] Detached leaf water loss rate: Leaves were cut off with sharp scissors, and the petioles were immediately removed to prevent errors caused by rapid water loss from the petiole wound. Three technical replicates were set up for each sample. After sampling, the leaves were immediately weighed on a balance to obtain the initial fresh weight (W1). Time nodes were set at 0, 1, 2, 3, 4, 5, and 6 hours, and the leaf weight (W2) was measured every hour. Detached leaf water loss rate = (W1-W2) / W1×100%. After obtaining the data, the time-water loss rate curve was plotted using GraphPad Prism 8 software, with the horizontal axis representing time and the vertical axis representing the cumulative water loss rate.

[0078] (III) Results and Analysis

[0079] 1. CDS sequence structure-level gene cloning of the StWRKY61 gene

[0080] The StWRKY61 gene was successfully isolated from the potato variety "Atlantic" using molecular cloning technology. Sequence analysis showed that the gene is 2587 bp in length, with a coding sequence (CDS) of 786 bp encoding 261 amino acids, as shown in SEQ ID NO. 1-2. Agarose gel electrophoresis results showed that the PCR amplification product appeared as a single bright band at the expected size (786 bp). Figure 1 This indicates that the StWRKY61 gene coding sequence with high specificity and integrity was successfully obtained, providing an accurate and reliable gene source for subsequent functional studies and vector construction. This result confirms the effectiveness of the designed primers and that no non-specific amplification was introduced during the cloning process, ensuring the accuracy of subsequent research.

[0081] The coding sequence (CDS) of the StWRKY61 gene, SEQ ID NO.1: ATGGATACAAGTTTGGGAGACAATAGTTTTTCCATTGACCTTAACACAAACCTCTCTTTGCACAACACCAATACAAGTCCGCGCGAGACGTTGGATGAAGAATTGATTAGGATGAGAGAGGAGAACAAGAAGTTAGTAACAATGCTCACAAATTTGTGTGAGAACTACAATTCCTTGCAAACTCACCTAATTGAGTTGCTGCAAAAATACTCTACTCATAATGAAGAGGACAATTCCAATTTATTATTACCAAGGAAAAGAAAAGCTGAAGAAGAGTGTTGTGTGAATAATTTTGAAGAAGCATCACCAAAGAGGCCAAGAGAAATCACAACCAATGTTTCAACTGTTTGTGTCAAAACTAATCGATCCGATCAAACCTCAGTGGTGAAGGATGGATATAACTGGAGAAAATATGGTCAAAAAGTTACAAGAGATAACCCTTATCCTAGAGCATATTATAAGTGTTCATTTGCACCAACATGCCCAGTCAAGAAGAAGGTACAAAGAAGCATTGAAGATCCATCAATTTTAGTAGCTGTATATGAAGGAGAACACAACCATCCTCATCCATCCCAAACTGAAATAACAGTACCATTAGTCAACCAAGATGTTACAACAGATCCAACATTTTCTAACAAATTCATGGAAGATATTGACACAAATTCATTACAACAACATTTAGTTCAACAAATGGCTTCTTCCTTAACGAGTAGCCCTACTTTCACAGCTGCAGTTGCTGCAGCCATCTCTGGAAAAATTTTCGAATATGATTTGCCTTTCAAATAA;

[0082] Protein sequence encoded by StWRKY61 gene: SEQ ID NO.2: MDTSLGDNSFSIDLNTNLSLHNTNTSPRETLDEELIRMREENKKLVTMLTNLCENYNSLQTHLIELLQKYSTHNEEDNSNLLLPRKRKAEEECCVNNFEEASPKRPREITTNVSTVCVKTNRSDQTSVV KDGYNWRKYGQKVTRDNPYPRAYYKCSFAPTCPVKKKVQRSIEDPSILVAVYEGEHNHPHPSQTEITVPLVNQDVTTDPTFSNKFMEDIDTNSLQQHLVQQMASSLTSSPTFTAAVAAAISGKIFEYDLPFK*.

[0083] 2. Construction of overexpression vectors

[0084] To investigate the function of StWRKY61, its CDS sequence was successfully constructed into the plant overexpression vector pC2300S-GFP. The recombinant plasmid was verified by double enzyme digestion (KpnⅠ / BamhⅠ). Agarose gel electrophoresis results showed that the digestion products exhibited a linearized vector band and a target gene band of approximately 786 bp at the expected positions. Figure 2 Undigested plasmids, on the other hand, appear as a single supercoiled band. Figure 2 This result clearly confirms that the StWRKY61 gene has been accurately inserted into the multiple cloning site of the vector, and that the vector construction was correct. The successful construction of this recombinant plasmid lays a solid foundation for subsequent genetic transformation and gene function verification.

[0085] 3. Genetic transformation process

[0086] The constructed StWRKY61 overexpression vector was introduced into potato mini-tuber explants using Agrobacterium-mediated genetic transformation. After co-culture, screening, and regeneration, resistant plants were successfully obtained. Figure 3 (A) Figure 3 Image A clearly demonstrates the complete regeneration process from callus formation (a), resistant bud differentiation (b), to fully rooted plantlets (c). To further confirm the transgenic plants, genomic DNA was extracted from the resistant seedlings for PCR identification.

[0087] The results showed that the transgenic lines (OE-1, OE-2, OE-3) were all able to amplify a specific band of 676 bp consistent with the positive control. Figure 3 The positive control (B) showed no such band, while the negative control (-) did not. This result strongly demonstrates that the StWRKY61 gene has been successfully integrated into the potato genome, resulting in multiple independent transgenic lines.

[0088] 4. Analysis of StWRKY61 expression levels in transgenic plants

[0089] The transcriptional level of StWRKY61 in wild-type (WT) and three transgenic lines (OE-1, OE-2, OE-3) was quantitatively analyzed using qRT-PCR. Results are as follows: Figure 4 As shown, compared with WT, the relative expression level of StWRKY61 in all transgenic lines showed a highly significant increase. This confirms at the transcriptional level that the exogenously introduced StWRKY61 gene was efficiently expressed in transgenic plants, indicating that a StWRKY61 overexpression system was successfully constructed. This result provides crucial molecular biological evidence for the subsequent observation of a series of drought-enhanced phenotypes, demonstrating that phenotypic differences do indeed stem from the overexpression of the target gene.

[0090] 5. Phenotypic comparison and physiological changes of transgenic plants under natural drought stress

[0091] After natural drought stress treatment, wild-type (WT) plants exhibited severe wilting and yellowing leaves, typical symptoms of drought damage, while the three overexpression lines (OE) consistently maintained good plant uprightness and green leaves. Figure 5 (A). Further quantification of this resistance difference was achieved through the determination of relevant physiological indicators: the relative leaf water content (RWC) of the overexpression line was significantly higher than that of the wild type. Figure 5 The presence of B indicates that it has a stronger water retention capacity; meanwhile, experiments on the water loss rate of detached leaves show that the water loss rate of the transgenic lines is significantly slower than that of the wild type. Figure 5 (C). These phenotypic and physiological data together demonstrate that overexpression of StWRKY61 can effectively enhance the water retention capacity of potato plants under drought conditions, thereby significantly improving their drought resistance.

[0092] 6. Changes in relevant physiological indicators of transgenic plants under PEG6000 simulated drought treatment

[0093] Under drought stress simulated by PEG6000, the transgenic plants again showed significant advantages. Phenotypic, the growth inhibition of the overexpression lines was significantly less than that of the severely wilted wild type. Figure 6 (A). Statistical analysis of growth indicators ( Figure 6Figure B shows that under normal conditions, there were no significant differences in growth indicators among the lines; however, after PEG6000 stress, the overexpressing lines (especially OE-2) were significantly superior to the wild type in key indicators such as plant height, root length, and fresh weight of stems and leaves. This fully demonstrates that overexpression of StWRKY61 not only improves the survival ability of plants under drought stress but also ensures the accumulation of biomass, especially promoting root development, which is crucial for water absorption in arid environments. This confirms the important role of this gene in positively regulating potato drought resistance from multiple dimensions.

[0094] After treatment with PEG6000, the transgenic lines overexpressing StWRKY61 showed greater growth in plant height and root extension than the wild type, indicating that it positively regulates potato drought resistance.

[0095] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. WRKY61 Genes, WRKY61 protein or containing WRKY61 The application of recombinant gene vectors, expression cassettes, transgenic cell lines, or recombinant bacteria in improving the drought resistance of potatoes is characterized by, overexpression WRKY61 Genes may increase the level of WRKY61 protein, thereby improving the drought resistance of potatoes; The WRKY61 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the amino acid sequence of the WRKY61 protein is shown in SEQ ID NO.

2.

2. A method for improving the drought resistance of potatoes, characterized in that, overexpression WRKY61 Genes may increase the level of WRKY61 protein, thereby improving the drought resistance of potatoes; The WRKY61 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the amino acid sequence of the WRKY61 protein is shown in SEQ ID NO.

2.

3. WRKY61 Genes, WRKY61 protein or containing WRKY61 The application of recombinant gene vectors, expression cassettes, transgenic cell lines, or recombinant bacteria in the cultivation of new potato varieties with strong drought resistance is characterized by, The WRKY61 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the amino acid sequence of the WRKY61 protein is shown in SEQ ID NO.

2.

4. A method for breeding new potato varieties with strong drought resistance, characterized in that, Includes the following steps: Will WRKY61 Genes are introduced into the target plant, thus obtaining the desired result; WRKY61 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

5. The method according to claim 4, characterized in that, The WRKY61 Genes through containing WRKY61 Gene recombinant vectors, expression cassettes, transgenic cell lines, or recombinant bacteria are introduced into target plants.

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