Application of zmbzip6 protein and its encoding gene in regulating plant tolerance to salt stress

Abstract

This invention discloses the application of the ZmbZIP6 protein and its encoding gene in regulating plant waterlogging tolerance, belonging to the field of biotechnology. This invention is the first to discover... ZmbZIP6 The application of genes and their encoded proteins in regulating plant waterlogging tolerance provides a theoretical basis for molecular breeding and germplasm resource improvement related to waterlogging tolerance; further research has found that... ZmbZIP6 After the gene is knocked out, the plant's waterlogging tolerance decreases, ZmbZIP6 Overexpression of the gene leads to increased waterlogging tolerance in plants; therefore, it has good application prospects in the field of agricultural production.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to the application of the ZmbZIP6 protein and its encoding gene in regulating plant waterlogging tolerance. Background Technology

[0002] Traditional plant breeding methods are time-consuming and prone to unpredictable results. The rapid development of transgenic technology has provided an important pathway to improving crop yield, stress resistance, and disease resistance. Utilizing transgenic technology and transgenic plant materials to study gene function at the physiological, biochemical, and molecular levels provides theoretical support for the creation of new materials and molecular breeding, which has significant practical implications. Improving crop waterlogging tolerance through the development of new varieties is a major way to enhance crop resistance to extreme climatic conditions and has important practical significance for increasing crop yield.

[0003] Traditional maize cultivation methods are susceptible to extreme weather, especially waterlogging caused by frequent rainfall. Utilizing genetic engineering techniques to obtain maize varieties with improved waterlogging tolerance, thereby increasing maize yield, has become a modern approach to genetic breeding.

[0004] Currently, for ZmbZIP6 The role of genes in regulating plant waterlogging tolerance has not been reported. Summary of the Invention

[0005] The purpose of this invention is to provide the application of the ZmbZIP6 protein and its encoding gene in regulating plant waterlogging tolerance. This addresses the problem of the lack of an efficient ZmbZIP6 protein and its encoding gene for regulating plant waterlogging tolerance in existing technologies.

[0006] In a first aspect, the present invention provides the application of ZmbZIP6 protein in regulating plant waterlogging tolerance, wherein the ZmbZIP6 protein is selected from any of the following: A1) a protein with an amino acid sequence as shown in SEQ ID NO.1; A2) a protein with the same function obtained by substitution, deletion or addition of one or more amino acids compared with the amino acid sequence defined in A1); A3) a protein with the same function obtained by attaching a tag to the N-terminus and / or C-terminus of A1) or A2).

[0007] The ZmbZIP6 protein provided by this invention can be a natural, recombinant, or synthetic active polypeptide. The active polypeptide can be a naturally purified product, a chemically synthesized product, or a product produced from a prokaryotic host (e.g., Escherichia coli) or a eukaryotic host (e.g., yeast, higher plants) using recombinant technology.

[0008] In this invention (A3), the linkage can be achieved through direct peptide bond connection or through a linker, using methods conventional in the art. The tags include, but are not limited to: GST (glutathione thioredoxin) tag protein, Trx (thioredoxin) tag protein, His tag protein (His-tag), Flag tag protein, LacZ tag protein, GFP (green fluorescent protein), sfGFP (hyperfolded green fluorescent protein), and HA tag (hemagglutinin tag). Those skilled in the art can select appropriate tag proteins according to actual needs. The use of tags does not alter the function of the target protein (ZmbZIP6 protein); its purpose is for separation, purification, detection, or tracing. The tags can be separated from the target protein (ZmbZIP6 protein) using chemical cleavage methods or enzymatic methods known in the art (e.g., introducing protease cleavage sites and using TEV protease to remove the tag).

[0009] In some implementations, the application is achieved by upregulating or downregulating the content and / or activity of the ZmbZIP6 protein.

[0010] In some implementations, plants with improved waterlogging tolerance are cultivated by upregulating the content and / or activity of the ZmbZIP6 protein.

[0011] In some preferred embodiments, upregulating the content and / or activity of the ZmbZIP6 protein can be achieved by overexpressing the gene for the ZmbZIP6 protein. ZmbZIP6 This is achieved through genes.

[0012] In some implementations, plants with reduced waterlogging tolerance are bred by downregulating the content and / or activity of the ZmbZIP6 protein.

[0013] In some preferred embodiments, downregulating the content and / or activity of the ZmbZIP6 protein can be achieved by reducing the ZmbZIP6 protein gene (…). ZmbZIP6 The expression level of genes (e.g., knockout) ZmbZIP6 This is achieved through genes.

[0014] In a second aspect, the present invention provides the application of biomaterials in regulating plant waterlogging tolerance, wherein the biomaterials are selected from any of the following: B1) a nucleic acid molecule encoding the ZmbZIP6 protein described above; B2) a nucleic acid molecule that inhibits or reduces the expression of the gene encoding the ZmbZIP6 protein described above; B3) a recombinant vector containing either B1) or B2) a nucleic acid molecule; B4) a recombinant cell containing either B1) or B2) a nucleic acid molecule, or a recombinant cell containing a recombinant vector of B3); B5) a transgenic plant containing either B1) or B2) a nucleic acid molecule.

[0015] In some embodiments, B1) the nucleic acid molecule is selected from any of the following: C1) a nucleic acid molecule having the nucleotide sequence shown in SEQ ID NO.2; C2) a nucleic acid molecule that hybridizes with the nucleic acid molecule defined in C1) under stringent conditions and encodes the ZmbZIP6 protein described above; C3) a nucleic acid molecule that has more than 90% sequence identity with the nucleic acid molecule defined in C1) or C2) and encodes the ZmbZIP6 protein described above.

[0016] The nucleic acid molecules provided by this invention can be DNA, such as cDNA, genomic DNA, or recombinant DNA; or RNA, such as mRNA or hnRNA; and these nucleic acid molecules can usually be obtained by PCR amplification or artificial synthesis.

[0017] As used herein, the term "hybridization under stringent conditions" refers to the hybridization of two nucleic acid fragments under standard hybridization conditions as described in the section "Expression of Cloned Genes in E. coli" of *Molecular Cloning: A Laboratory Manual* (1989) (Cold Spring Lane Laboratory, New York, USA). Such conditions include hybridization in 6.0 × SSC at 45 °C, followed by washing in 2 × SSC at 50 °C. To select stringency, the salt concentration in the washing step can be chosen, for example, between 2.0 × SSC at 50 °C for low stringency and 2.0 × SSC at 50 °C for high stringency. Additionally, the temperature in the washing step can be varied between room temperature (approximately 22 °C) for low stringency and 65 °C for high stringency.

[0018] As used herein, the term “sequence identity” can be evaluated by the naked eye or by computer software (such as the software program described in Current Protocols in Molecular Biology by Ausubel et al. eds. (2007)). Molecules are identical at that position when positions in the compared sequences are occupied by the same bases or amino acids. Identity between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the identity between related sequences. “Sequence identity” of a polynucleotide or amino acid sequence with another sequence having a certain percentage (e.g., 90%, 95%, 98%, or 99%) means that when the sequences are aligned, that percentage of bases or amino acids are the same in the two compared sequences.

[0019] In some embodiments, B2) the nucleic acid molecule is sgRNA, and the target sequence of sgRNA includes at least one of sgRNA1 with a nucleotide sequence as shown in SEQ ID NO.3 and sgRNA2 with a nucleotide sequence as shown in SEQ ID NO.4.

[0020] In this invention, the inventors discovered that selecting the aforementioned specific target sequence results in high knockout efficiency.

[0021] In a third aspect, the present invention provides a method for cultivating plants with reduced waterlogging tolerance, the method comprising reducing the content and / or activity of the aforementioned ZmbZIP6 protein in the target plant to obtain a plant with lower waterlogging tolerance than the target plant.

[0022] In some implementations, reducing the content and / or activity of the ZmbZIP6 protein in the target plant is achieved by reducing the expression level of the gene encoding the ZmbZIP6 protein in the target plant.

[0023] In some implementations, reducing the expression level of the gene encoding the ZmbZIP6 protein in the target plant is achieved using a CRISPR / Cas9 system, which includes the aforementioned sgRNA.

[0024] In some implementation schemes, the method for cultivating plants with reduced waterlogging tolerance specifically includes the following steps: 1) constructing a DNA molecule encoding sgRNA into a Cas9 expression vector to obtain a CRISPR / Cas9 gene editing vector; 2) introducing the CRISPR / Cas9 gene editing vector into the target plant; 3) obtaining the desired gene editing vector through screening and identification. ZmbZIP6 Gene knockout transgenic plants are plants with reduced waterlogging tolerance.

[0025] In some preferred embodiments, in step 2), the introduction can be carried out by Agrobacterium-mediated transformation, specifically including the following steps: introducing the CRISPR / Cas9 gene editing vector into Agrobacterium to obtain recombinant Agrobacterium; infecting maize callus or explants with the recombinant Agrobacterium; and inducing and culturing the obtained positive callus or explants to obtain regenerated plants after identification.

[0026] In a fourth aspect, the present invention provides a method for improving the waterlogging resistance of a plant, the method comprising increasing the content and / or activity of the aforementioned ZmbZIP6 protein in the target plant to obtain a plant with higher waterlogging resistance than the target plant.

[0027] In some implementations, increasing the content and / or activity of the ZmbZIP6 protein in the target plant is achieved by increasing the expression level of the gene encoding the ZmbZIP6 protein in the target plant.

[0028] In some implementation schemes, increasing the expression level of the gene encoding the ZmbZIP6 protein in the target plant can be achieved by at least one of the following methods: 1) increasing the copy number of the gene encoding the ZmbZIP6 protein; 2) placing the gene encoding the ZmbZIP6 protein under the drive of a strong promoter for expression; 3) increasing the regulatory elements of the gene encoding the ZmbZIP6 protein to overexpress it, including enhancer elements, elements that improve mRNA stability, elements that improve translation efficiency, and / or elements that improve protein secretion; 4) codon optimization of the gene encoding the ZmbZIP6 protein.

[0029] In some implementation schemes, the method for cultivating plants with improved waterlogging tolerance specifically includes the following steps: 1) constructing a recombinant vector containing a nucleic acid molecule encoding the ZmbZIP6 protein; 2) introducing the constructed recombinant vector into the target plant; and 3) obtaining transgenic plants through screening and identification.

[0030] In some preferred embodiments, in step 2), the introduction can be carried out by Agrobacterium-mediated transformation, specifically including the following steps: introducing the recombinant vector into Agrobacterium to obtain recombinant Agrobacterium; infecting maize callus or explants with the recombinant Agrobacterium; and inducing and culturing the obtained positive callus or explants to obtain regenerated plants after identification.

[0031] It is understood that in step 3), the screening and identification methods are conventional methods in the field, such as screening and identification by PCR detection.

[0032] In some implementations, the plant includes corn.

[0033] The beneficial effects of this invention are: unlike the prior art, this invention is the first to discover... ZmbZIP6 The application of genes and their encoded proteins in regulating plant waterlogging tolerance provides a theoretical basis for molecular breeding and germplasm resource improvement related to waterlogging tolerance; further research has found that... ZmbZIP6 Knocking out the gene reduces the plant's tolerance to waterlogging. ZmbZIP6 Overexpression of the gene leads to increased waterlogging tolerance in plants; therefore, it has good application prospects in the field of agricultural production. Attached Figure Description

[0034] Figure 1 In Embodiment 2 of the present invention ZmbZIP6 Schematic diagram of gene targets; Figure 2A ZmbZIP6 in Embodiment 2 of the present invention ko1 Line sequencing results; Figure 2B ZmbZIP6 in Embodiment 2 of the present invention ko2 Line sequencing results; Figure 3 In the overexpression line of Example 3 of this invention ZmbZIP6 qPCR test results of the gene; Figure 4 ZmbZIP6 in Embodiment 4 of the present invention OE1 strain, ZmbZIP6 OE2 Phenotypic results of the strains, in which (a) the left side is ZmbZIP6 under normal growth. NT The right side shows a ZmbZIP6 corn plant under normal growth conditions. OE1 (b) shows maize plants with a bar of 5 cm; the left side of (b) shows ZmbZIP6 after 15 days of waterlogging stress treatment. NT The right side shows ZmbZIP6 maize plants after 15 days of waterlogging stress treatment. OE1 The corn plant strain, Bar=10cm; (c) The left side shows the ZmbZIP6 under normal growth. NT The right side shows a ZmbZIP6 corn plant under normal growth conditions. OE2 Maize plants of strain , Bar=5cm; (d) The left side of the image shows ZmbZIP6 after 15 days of waterlogging stress treatment. NT The right side shows ZmbZIP6 maize plants after 15 days of waterlogging stress treatment. OE2 The maize plant strain, Bar=10cm; (e) shows ZmbZIP6 after 15 days of waterlogging stress. OE1 Plant height bar chart; (f) shows ZmbZIP6 after 15 days of waterlogging stress. OE2 Plant height bar chart; Figure 5 ZmbZIP6 in Embodiment 4 of the present invention ko1 strain, ZmbZIP6 ko2 Phenotypic results of the strains, in which (a) the left side is ZmbZIP6 under normal growth. NT The right side shows a ZmbZIP6 corn plant under normal growth conditions. ko1 (b) shows maize plants with a bar of 5 cm; the left side of (b) shows ZmbZIP6 after 15 days of waterlogging stress treatment. NT The right side shows ZmbZIP6 maize plants after 15 days of waterlogging stress treatment. ko1 The corn plant strain, Bar=10cm; c) The left side of the image shows ZmbZIP6 under normal growth conditions. NT The right side shows a ZmbZIP6 corn plant under normal growth conditions. ko2 The maize plant strain, Bar=5cm; d) The left side of the image shows the result of waterlogging stress treatment for 15 days (ZmbZIP6). NT The right side shows ZmbZIP6 maize plants after 15 days of waterlogging stress treatment.ko2 The maize plant strain, Bar=10cm; e) is the ZmbZIP6 after 15 days of waterlogging stress. ko1 Plant height bar chart; f) ZmbZIP6 after 15 days of waterlogging stress. ko2 Plant height bar chart. Detailed Implementation

[0035] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0036] Experimental methods not specifically described in the examples are generally performed according to conventional experimental methods in the field of molecular biology, including but not limited to those described in *Molecular Cloning: A Laboratory Manual* by M.R. Green and *Molecular Biology* by Robert F. Weaver, or according to the experimental methods recommended by the reagent kit and instrument manufacturers. Unless otherwise specified, all reagents and biological materials used in the examples are commercially available.

[0037] Example 1 ZmbZIP6 Cloning of genes Designed for amplification ZmbZIP6 Primers for the gene CDS were used to perform PCR amplification using total DNA from leaves of maize inbred line KN5585 (extracted using the CTAB method) as a template. ZmbZIP6 The primer sequences for the gene CDS are shown below: ZmbZIP6-F:5′-ATGCACATGGCGCTATCGTC-3′ (SEQ ID NO.5); ZmbZIP6-R: 5′-GGATGCATCAGCCAGGCCAT-3′ (SEQ ID NO. 6).

[0038] The reaction system was as follows: 7.5 µl of 2×Taq Mster Mix, 1 µl of ZmbZIP6-F, 1 µl of ZmbZIP6-R, 1 µl of DNA, and 4.5 µl of ddH2O, for a total volume of 15 µl. The reaction program was as follows: 95 °C pre-denaturation for 5 min, 95 °C denaturation for 30 s, 58 °C annealing for 30 s, 72 °C extension for 60 s, repeated 34 times, followed by a final extension at 72 °C for 5 min. After the reaction was complete, the PCR product was recovered and, after correct sequencing, yielded... ZmbZIP6 Gene.

[0039] in, ZmbZIP6 The nucleotide sequence of the gene is shown in SEQ ID NO.2, and the amino acid sequence of the ZmbZIP6 protein is shown in SEQ ID NO.1.

[0040] Example 2 ZmbZIP6 Obtaining and identifying gene knockout plants 2.1 Construction of gene editing vectors Gene target design was performed using the website (http: / / cbi.hzau.edu.cn / crispr / ), resulting in two sgRNAs: sgRNA1: 5′-TCGTATCGTCAGCCGTGCAGCGG-3′ (SEQ ID NO.3) and sgRNA2: 5′-CCGCGCCGAACGCTGCCGACGGG-3′ (SEQ ID NO.4). Their target sites are described in [link to relevant documentation]. Figure 1 (Target1 and Target2, respectively) The ZmU6 promoters of the first and second targets were amplified using the designed pU6F1 / pU6R primer pair and pU6F2 / pU6R primer pair, respectively; simultaneously, Target1-F and Target2-F sequences with added homologous arms were designed using two sgRNA sequences, and Target1+sgRNA1 and Target2+sgRNA2 fragments were amplified using the designed Target1-F / gRR0 and Target2-F / gRR1, respectively; then, overlap PCR amplification was performed using pU6F1 / gRR0 and pU6F2 / gRR1, respectively. The amplification reaction yielded ZmU6-Target1-sgRNA1 and ZmU6-Target2-sgRNA2 fragments, respectively. These fragments, amplified by overlapping PCR, were then recombined into the HindIII-digested CPB-ZmUbi-hspCas9 vector (see Chinese patent document CN116574754A). Furthermore, the recombinant vectors were sequenced using CRISPR-F / CRISPR-R vector detection primer pairs. After successful sequencing, two gene-editing vectors were obtained. The sequences of the primer pairs used in the construction of the gene-editing vectors are shown in Table 1 below.

[0041] Table 1 Primer Sequences

[0042] 2.2 Obtaining Gene-Edited Plants The two gene-editing vectors were transformed into the maize inbred line KN5585 via Agrobacterium-mediated genetic transformation. The genetic transformation was carried out by Weimi Biotechnology Co., Ltd., and the resulting transgenic plants were designated as generation T0.

[0043] Genomic DNA was extracted from the above-mentioned transgenic T0 generation plants, the target fragment was amplified, and then Sanger sequencing was performed to analyze the type of gene editing. The results are as follows: Figure 2A-2B As shown.

[0044] As can be seen from 2A-2B, ZmbZIP6 ko1 The strain editing type involves an 18bp deletion at the target site, resulting in the deletion of amino acids 21-27; ZmbZIP6 ko2 The edited strain involved a 17bp deletion at the target site, resulting in a frameshift mutation that prematurely terminated translation, leading to a truncated protein of 90 amino acids. These results demonstrate that this invention can efficiently obtain… ZmbZIP6 Genetically modified plants with gene knockout.

[0045] Example 3 ZmbZIP6 Obtaining and identifying gene-overexpressing plants 3.1 Construction of overexpression vectors The amplification obtained in Example 2 ZmbZIP6 The gene fragment was ligated into the pBWA(V)HS vector, and after sequencing verification, pBWA(V)HS- was obtained. ZmbZIP6 Recombinant vector.

[0046] 4.2 Obtaining overexpression plants The above pBWA(V)HS- ZmbZIP6 The recombinant vector was genetically transformed into the maize inbred line KN5585 via Agrobacterium-mediated transformation. The genetic transformation was carried out by Weimi Biotechnology Co., Ltd., and the resulting transgenic plants were designated as generation T0.

[0047] RNA was extracted from the above-mentioned transgenic T0 generation plants, and cDNA was obtained by reverse transcription. The cDNA was then detected using qPCR to obtain the cDNA from the T0 generation transgenic plants. ZmbZIP6 The relative expression levels of genes. The primers used for qPCR are as follows: ZmbZIP6- qPCR-F: 5′-CAGCCACCATGTCGTATCGT-3′ (SEQ ID NO. 16); ZmbZIP6- qPCR-R: 5′-GCCGCTGCACTCCTAGTAT-3′ (SEQ ID NO. 17); The reaction mixture is as follows: 10 µl of 2×ChamQ Blue Universal SYBR qPCR Master Mix. ZmbZIP6- qPCR-F 0.4µl, ZmbZIP6- 0.4 µl of qPCR-R, 2 µl of cDNA, and 7.2 µl of ddH2O were used, with a total volume of 20 µl. The reaction program was as follows: 95 °C pre-denaturation for 5 min, 95 °C denaturation for 20 s, 60 °C annealing for 20 s, and 72 °C extension for 20 s, for 40 cycles. -△△ct The method performs data analysis and plots using GraphPad Prism8, and the results are as follows: Figure 3 As shown.

[0048] from Figure 3 As can be seen from this, compared to wild-type plants (ZmbZIP6) NT ), overexpression line (ZmbZIP6) OE1 ZmbZIP6 OE2 )middle, ZmbZIP6 The relative expression level of the gene was significantly increased; the above results indicate that the present invention can efficiently obtain... ZmbZIP6 Transgenic plants with overexpressed genes.

[0049] Example 4 ZmbZIP6 Gene knockout and ZmbZIP6 Phenotypic identification of gene overexpression plants The ZmbZIP6 obtained in Example 2 above ko1 strain, ZmbZIP6 ko2 The strain, ZmbZIP6 obtained in Example 3 OE1 strain, ZmbZIP6 OE2 The T0 generation plants of the strain were hybridized to obtain the T1 generation plants. The number of days in different growth stages of the T1 generation plants was counted, and the results are shown in Table 2 below.

[0050] Table 2 Statistics on reproductive period

[0051] As can be seen from Table 1, overexpression ZmbZIP6 The gene has almost no effect on the entire growth period of the plant, while the knockout... ZmbZIP6 Genetic delays can lead to a lag in the entire growth period of plants.

[0052] Furthermore, in a plastic storage box measuring 66cm in length, 42cm in width, and 40cm in height, ZmbZIP6 seedlings grown to the three-leaf-one-heart stage were respectively... NT strain, ZmbZIP6 OE1 strain, ZmbZIP6 OE2strain, ZmbZIP6 ko1 strain, ZmbZIP6 ko2 The strains were subjected to a 15-day waterlogging stress treatment to investigate... ZmbZIP6 The effects of genes on stain resistance are as follows: Figure 4 and 5 As shown.

[0053] from Figure 4 As can be seen from the data, after 15 days of waterlogging stress treatment, ZmbZIP6... OE1 strains and ZmbZIP6 OE2 The growth of the strain was significantly better than that of ZmbZIP6. NT Good strain ( Figure 4 (b, 4d), in both treatments (normal growth and waterlogging stress treatment), ZmbZIP6 NT The average plant height of the strains was 38.10 ( Figure 4 e) and 38.09 ( Figure 4 f), ZmbZIP6 OE1 The average plant height of the strain was 41.20 mm. Figure 4 e), ZmbZIP6 OE2 The average plant height of the strain was 41.53 cm. Figure 4 f); the results showed that overexpression ZmbZIP6 After gene modification, the plant's waterlogging tolerance is significantly improved.

[0054] from Figure 5 As can be seen from the data, after 15 days of waterlogging stress treatment, ZmbZIP6... ko1 strains and ZmbZIP6 ko2 The growth of the strain was significantly better than that of ZmbZIP6. NT Different strains ( Figure 5 (b, 5d), in the two treatment groups (normal growth and waterlogging stress treatment), ZmbZIP6 NT The average plant height of the strains was 38.09 ( Figure 5 e) and 38.11 Figure 5 f), ZmbZIP6 ko1 The average plant height of the strain was 27.62 mm. Figure 5 e), ZmbZIP6 ko2 The average plant height of the strain was 27.66 ( Figure 5 f); the results showed that knockout ZmbZIP6 After gene modification, the plant's waterlogging tolerance is significantly reduced.

[0055] In summary, this invention is the first to discover... ZmbZIP6 The application of genes and their encoded proteins in regulating plant waterlogging tolerance provides a theoretical basis for molecular breeding and germplasm resource improvement related to waterlogging tolerance.

[0056] The embodiments described above are merely illustrative of implementation methods of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of ZmbZIP6 protein in regulating plant waterlogging tolerance, characterized in that, The ZmbZIP6 protein is selected from any of the following: A1) A protein with the amino acid sequence shown in SEQ ID NO.1; A3) Proteins with the same function obtained by attaching tags to the N-terminus and / or C-terminus of A1) or A2); The plant in question is corn; The regulation mentioned is a positive regulation.

2. The application of biomaterials in regulating plant waterlogging tolerance, characterized in that, The biomaterial is selected from any one of the following: B1) A nucleic acid molecule encoding the ZmbZIP6 protein of claim 1; B3) A recombinant vector containing the nucleic acid molecules described in B1); B4) Recombinant cells containing the nucleic acid molecules described in B1), or recombinant cells containing the recombinant vector described in B3); The plant in question is corn; The regulation mentioned is a positive regulation.

3. The application according to claim 2, characterized in that, B1) The nucleic acid molecule is selected from any of the following: C1) Nucleic acid molecules with nucleotide sequences as shown in SEQ ID NO.

2.

4. A method for cultivating plants with improved waterlogging tolerance, characterized in that, The method includes increasing the content of the ZmbZIP6 protein of claim 1 in the target plant to obtain a plant with higher waterlogging resistance than the target plant; The plant in question is corn.

5. The method according to claim 4, characterized in that, The improvement of the ZmbZIP6 protein content in the target plant according to claim 1 is achieved by increasing the expression level of the gene encoding the ZmbZIP6 protein in the target plant.

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