Application of ZmbZIP6 protein and coding gene thereof in regulation and control of plant waterlogging tolerance

Abstract

The invention discloses application of ZmbZIP6 protein and a coding gene thereof in regulation and control of plant waterlogging tolerance, and belongs to the technical field of biology. The application of the ZmbZIP6 gene and the protein coded by the ZmbZIP6 gene in regulation and control of plant waterlogging tolerance is found for the first time, and a theoretical basis is provided for molecular breeding and germplasm resource improvement related to waterlogging tolerance; further research finds that after the ZmbZIP6 gene is knocked out, the waterlogging tolerance of the plant is reduced, and after the ZmbZIP6 gene is over-expressed, the waterlogging tolerance of the plant is improved; therefore, the method has a good application prospect in the field of agricultural production.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and particularly relates to application of ZmbZIP6 protein and its coding gene in regulating plant tolerance to waterlogging. BACKGROUND

[0002] Traditional plant breeding methods are time-consuming and have unpredictable results, and the rapid development of transgenic technology provides an important way for improving crop yield, stress resistance and disease resistance. Using transgenic technology and transgenic plant materials to study the function of genes from the physiological and biochemical and molecular levels provides theoretical support for creating new materials and molecular breeding, which has great practical significance. Through the cultivation of new varieties, the tolerance of crops to waterlogging is improved, which is a major way to improve the resistance of crops to extreme weather conditions and has important practical significance for improving crop yield.

[0003] Traditional methods of corn cultivation are easily affected by extreme weather, especially by waterlogging disasters caused by frequent rainfall. Using genetic engineering technology to obtain varieties with improved genetic tolerance to waterlogging and to improve corn yield is a way of modern genetic breeding.

[0004] At present, the function of the gene in regulating plant tolerance to waterlogging has not been reported. ZmbZIP6 The function of the gene in regulating plant tolerance to waterlogging has not been reported. SUMMARY

[0005] The application aims to provide application of ZmbZIP6 protein and its coding gene in regulating plant tolerance to waterlogging, and solve the problem of lack of ZmbZIP6 protein and its coding gene for efficiently regulating plant tolerance to waterlogging in the prior art.

[0006] In a first aspect, the application provides application of ZmbZIP6 protein in regulating plant tolerance to waterlogging, wherein the ZmbZIP6 protein is selected from any one 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 several amino acids compared with the amino acid sequence defined in A1); and A3) a protein with the same function obtained by connecting a tag to the N terminal and / or C terminal of A1) or A2).

[0007] The ZmbZIP6 protein provided by the application can be a natural, recombinant or synthetic active polypeptide, which can be a natural purified product, a chemically synthesized product, or a product produced from a prokaryotic host (such as E. coli) or a eukaryotic host (such as yeast, higher plants) using a recombinant technology.

[0008] In A3) of the present application, the connection can be direct connection through a peptide bond or connection through a linker, and the method of connection is a method conventional in the art. Among them, the tag includes but is not limited to: GST (glutathione S-transferase) tag protein, Trx (thioredoxin) tag protein, His tag protein (His-tag), Flag tag protein, LacZ tag protein, GFP (green fluorescent protein), sfGFP (super-fold green fluorescent protein), HA tag (hemagglutinin tag). A person skilled in the art can select a suitable tag protein according to the actual use needs. The use of the tag does not change the function of the target protein (ZmbZIP6 protein), and the purpose is to separate, purify, detect or track. The tag can be separated from the target protein (ZmbZIP6 protein) by chemical cleavage method or enzymatic method known in the art (such as introducing a protease cleavage site to remove the tag using TEV protease).

[0009] In some embodiments, the application is achieved by up-regulating or down-regulating the content and / or activity of the ZmbZIP6 protein.

[0010] In some embodiments, the application is achieved by up-regulating the content and / or activity of the ZmbZIP6 protein to cultivate plants with improved salt tolerance.

[0011] In some preferred embodiments, up-regulating the content and / or activity of the ZmbZIP6 protein can be achieved by over-expressing the gene of the ZmbZIP6 protein. ZmbZIP6 Gene).

[0012] In some embodiments, the application is achieved by down-regulating the content and / or activity of the ZmbZIP6 protein to cultivate plants with reduced salt tolerance.

[0013] In some preferred embodiments, down-regulating the content and / or activity of the ZmbZIP6 protein can be achieved by reducing the expression amount of the gene of the ZmbZIP6 protein (for example, knocking out the gene of the ZmbZIP6 protein). ZmbZIP6 Gene). ZmbZIP6 Gene).

[0014] In a second aspect, the present application provides the use of a biological material in regulating the salt tolerance of plants, wherein the biological material is selected from any one of the following: B1) a nucleic acid molecule encoding the above-mentioned ZmbZIP6 protein; B2) a nucleic acid molecule inhibiting or reducing the expression of the gene encoding the above-mentioned ZmbZIP6 protein; B3) a recombinant vector comprising the nucleic acid molecule of B1) or B2); B4) a recombinant cell comprising the nucleic acid molecule of B1) or B2), or a recombinant cell comprising the recombinant vector of B3); B5) a transgenic plant comprising the nucleic acid molecule of B1) or B2).

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

[0016] The above-mentioned nucleic acid molecule provided by the present application can be DNA, such as cDNA, genomic DNA or recombinant DNA; or RNA, such as mRNA or hnRNA, etc. The nucleic acid molecule can be obtained by PCR amplification or artificial synthesis.

[0017] As used herein, the term "hybridizes under stringent conditions" means that two nucleic acid molecule fragments hybridize to each other under standard hybridization conditions as described in the section "Expression of cloned genes in E. coli" of Sambrook et al. Molecular Cloning: A Laboratory Manual (1989) (Cold Spring Harbor Laboratory Press, New York, USA). Such conditions are, for example, hybridization in 6.0 x SSC at 45°C followed by a washing step in 2 x SSC at 50°C. In order to select the stringency, the salt concentration in the washing step can be chosen, for example, between 2.0 x SSC at 50°C for low stringency and 2.0 x SSC at 50°C for high stringency. In addition, the temperature in the washing step can vary between about room temperature of about 22°C for low stringency and 65°C for high stringency.

[0018] As used herein, the term "sequence identity" can be assessed by eye or by computer software, such as the software programs described in Ausubel et al. eds. (2007) in Current Protocols in Molecular Biology. When a position in compared sequences is occupied by the same base or amino acid, then the molecules are identical at that position. A degree of identity between two or more sequences can be expressed as a percentage (%) which can be used to assess the identity between related sequences. A polynucleotide sequence or an amino acid sequence has a certain percentage (e.g. 90%, 95%, 98% or 99%) of "sequence identity" to another sequence if that percentage of bases or amino acids are the same in the two sequences when aligned.

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

[0020] In the present application, the inventors have found that the selection of the specific target sequence has a higher knockout efficiency.

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

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

[0023] In some embodiments, the reduction of the expression level of the gene encoding the ZmbZIP6 protein in the plant of interest is achieved by using a CRISPR / Cas9 system, which comprises the sgRNA.

[0024] In some embodiments, the method for breeding a plant with reduced tolerance to waterlogging specifically comprises the following steps: 1) constructing a DNA molecule encoding the sgRNA into a Cas9 expression vector to obtain a CRISPR / Cas9 gene editing vector; 2) introducing the CRISPR / Cas9 gene editing vector into the plant of interest; and 3) obtaining a transgenic plant with reduced tolerance to waterlogging through screening and identification. ZmbZIP6 The transgenic plant with gene knockout is a plant with reduced tolerance to waterlogging.

[0025] In some preferred embodiments, in step 2), the introduction can be achieved by Agrobacterium-mediated method, which specifically comprises the following steps: introducing the CRISPR / Cas9 gene editing vector into Agrobacterium to obtain a recombinant Agrobacterium; infecting the callus or explant of the corn with the recombinant Agrobacterium; and inducing the obtained positive callus or explant to obtain a regenerated plant through identification.

[0026] In a fourth aspect, the present application provides a method for breeding a plant with improved tolerance to waterlogging, the method comprising increasing the content and / or activity of the ZmbZIP6 protein in a plant of interest to obtain a plant with higher tolerance to waterlogging than the plant of interest.

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

[0028] In some embodiments, the expression of the gene encoding the ZmbZIP6 protein in the plant of interest can be increased by at least one of the following ways: 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, the regulatory elements including enhancer elements, elements for improving mRNA stability, elements for enhancing translation efficiency, and / or elements for enhancing protein secretion; and 4) codon optimization of the gene encoding the ZmbZIP6 protein.

[0029] In some embodiments, the method for breeding a plant with improved salt tolerance comprises the following steps: 1) constructing a recombinant vector comprising a nucleic acid molecule encoding the ZmbZIP6 protein; 2) introducing the constructed recombinant vector into a plant of interest; and 3) obtaining a transgenic plant through screening and identification.

[0030] In some preferred embodiments, in step 2), the introduction can be achieved by Agrobacterium-mediated method, which comprises the following steps: introducing the recombinant vector into Agrobacterium to obtain a recombinant Agrobacterium; infecting a callus or an explant of the corn with the recombinant Agrobacterium; and inducing the obtained positive callus or explant to obtain a regenerated plant through identification.

[0031] It can be understood that, in step 3), the screening and identification method is a conventional method in the art, for example, screening and identification can be achieved by PCR detection.

[0032] In some embodiments, the plant comprises corn.

[0033] The present application has the following beneficial effects: different from the prior art, the present application first discovers that ZmbZIP6 the gene and the protein encoded thereby are used for regulating the salt tolerance of plants, which provides a theoretical basis for molecular breeding and germplasm improvement related to salt tolerance; and further research finds that ZmbZIP6 knocking out the gene leads to reduced salt tolerance of the plant, and ZmbZIP6 overexpressing the gene leads to improved salt tolerance of the plant; therefore, the present application has good application prospects in the field of agricultural production. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 The following is a schematic diagram of the gene target in Example 2 of the present application; ZmbZIP6 The following is a schematic diagram of the gene target in Example 2 of the present application; Figure 2A The following is a schematic diagram of the gene target in Example 2 of the present application; ko1 The following is a schematic diagram of the gene target in Example 2 of the present application; Figure 2B The following is a schematic diagram of the gene target in Example 2 of the present application; ko2 The following is a schematic diagram of the gene target in Example 2 of the present application; Figure 3 For the overexpression strain in Example 3 of the present invention ZmbZIP6 qPCR detection results of genes; Figure 4 ZmbZIP6 in Example 4 of the present invention OE1 strain, ZmbZIP6 OE2 Phenotypic test results of the strains, where the left side in (a) shows ZmbZIP6 under normal growth NT The right side shows the normal growth of ZmbZIP6 corn plants. OE1 Corn plants of different strains, Bar=5cm; (b) on the left is ZmbZIP6 after 15 days of waterlogging stress treatment NT The right side shows the ZmbZIP6 corn plant after 15 days of waterlogging stress treatment. OE1 Corn plant of the strain, Bar=10cm; the left side of (c) shows ZmbZIP6 under normal growth NT The right side shows the normal growth of ZmbZIP6 corn plants. OE2 Corn plants of different strains, Bar=5cm; (d) on the left is ZmbZIP6 after 15 days of waterlogging stress treatment NT The right side shows the ZmbZIP6 corn plant after 15 days of waterlogging stress treatment. OE2 Corn plants of the same strain, Bar=10cm; (e) ZmbZIP6 after 15 days of waterlogging stress OE1 Plant height bar graph; (f) ZmbZIP6 after 15 days of waterlogging stress OE2 Histogram of plant height of strains; Figure 5 ZmbZIP6 in Example 4 of the present invention ko1 strain, ZmbZIP6 ko2 Phenotypic test results of the strains, where the left side in (a) shows ZmbZIP6 under normal growth NT The right side shows the normal growth of ZmbZIP6 corn plants. ko1 Corn plants of different strains, Bar=5cm; (b) on the left is ZmbZIP6 after 15 days of waterlogging stress treatment NT The right side shows the ZmbZIP6 corn plant after 15 days of waterlogging stress treatment. ko1 Corn plant of the strain, Bar=10cm; c) The left side shows ZmbZIP6 under normal growth NT The right side shows the normal growth of ZmbZIP6 corn plants. ko2 Corn plants of different strains, Bar=5cm; d) The left side shows ZmbZIP6 after 15 days of waterlogging stress treatment. NT The right side shows the ZmbZIP6 corn plant after 15 days of waterlogging stress treatment.ko2 Corn plants of the same strain, Bar=10cm; e) ZmbZIP6 after 15 days of waterlogging stress ko1 Plant height bar graph; f) ZmbZIP6 after 15 days of waterlogging stress ko2 Histogram of plant height of strains. DETAILED DESCRIPTION

[0035] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0036] Experimental procedures in the examples, where specific conditions are not specified, were generally performed in accordance with conventional methods in molecular biology, including but not limited to those described in M.R. Green's Molecular Cloning: A Laboratory Manual and Robert F. Weaver's Molecular Biology, or according to the recommendations of kit and instrument manufacturers. Unless otherwise specified, reagents and biological materials used in the examples were commercially available.

[0037] Example 1 ZmbZIP6 Gene cloning Designed for amplification ZmbZIP6 The primers of gene CDS were used for PCR amplification using the total DNA of leaves of maize inbred line KN5585 (which was extracted by CTAB method) as 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: 2×Taq Master Mix 7.5µl, ZmbZIP6-F 1µl, ZmbZIP6-R 1µl, DNA 1µl, ddH2O 4.5µl, total system 15µl; the reaction procedure was as follows: 95℃ pre-denaturation for 5min, 95℃ denaturation for 30s, 58℃ annealing for 30s, 72℃ extension for 60s, 34 cycles, and 72℃ extension for another 5min; after the reaction was completed, the PCR product was recovered and sequenced correctly to obtainZmbZIP6 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 according to the website (http: / / cbi.hzau.edu.cn / crispr / ), and two sgRNAs were finally obtained, namely sgRNA1: 5′-TCGTATCGTCAGCCGTGCAGCGG-3′ (SEQ ID NO. 3) and sgRNA2: 5′-CCGCGCCGAACGCTGCCGACGGG-3′ (SEQ ID NO. 4). The target sites are shown in Figure 1 (Target1 and Target2, respectively), the designed pU6F1 / pU6R primer pair and pU6F2 / pU6R primer pair were used to amplify the ZmU6 promoter of the first and second targets respectively; at the same time, the Target1-F and Target2-F sequences with homology arms were designed using two sgRNA sequences, and the designed Target1-F / gRR0 and Target2-F / gRR1 were used to amplify the Target1+sgRNA1 and Target2+sgRNA2 fragments respectively; then, overlapping PCR amplification was performed using pU6F1 / gRR0 and pU6F2 / gRR1 respectively. The overlapping PCR amplification reaction was performed to obtain ZmU6-Target1-sgRNA1 and ZmU6-Target2-sgRNA2 fragments, respectively. The above overlapping PCR amplification reaction fragments were homologously recombined into the HindIII-digested CPB-ZmUbi-hspCas9 vector (for this vector, see Chinese patent document CN116574754A). Furthermore, the obtained recombinant vectors were sequenced using the CRISPR-F / CRISPR-R vector detection primer pair. After correct sequencing, two gene editing vectors were obtained. The sequences of the primer pairs used in the construction of the above gene editing vectors are shown in Table 1 below.

[0041] Table 1 Primer sequences

[0042] 2.2 Obtaining gene-edited plants The above two gene editing vectors were respectively transformed into the maize inbred line KN5585 through Agrobacterium-mediated genetic transformation, wherein the genetic transformation was completed by Weimi Biotechnology Co., Ltd., and transgenic plants were obtained, which were recorded as T0 generation.

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

[0044] As can be seen from 2A-2B, ZmbZIP6 ko1 The editing type of the strain is an 18bp base deletion at the target site, and the above base deletion causes the deletion of amino acids 21-27; ZmbZIP6 ko2 The editing type of the strain is a 17bp base deletion at the target site, which causes a frameshift mutation, resulting in premature termination of translation and the formation of a truncated protein consisting of 90 amino acids; the above results show that the present invention can efficiently obtain ZmbZIP6 Gene knockout transgenic plants.

[0045] Example 3 ZmbZIP6 Obtaining and identifying gene-overexpressing plants 3.1 Construction of overexpression vector The amplified ZmbZIP6 The gene fragment was connected to the pBWA(V)HS vector and verified by sequencing to obtain pBWA(V)HS- 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 genetic transformation, wherein the genetic transformation was completed by Weimi Biotechnology Co., Ltd., and transgenic plants were obtained, which were recorded as T0 generation.

[0047] The RNA of the above-mentioned transgenic T0 plants was extracted and reverse transcribed to obtain cDNA. Then, the qPCR detection method was used to obtain the cDNA of the T0 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 system is as follows: 2×ChamQ Blue Universal SYBR qPCR Master Mix 10µl, ZmbZIP6- qPCR-F 0.4µl, ZmbZIP6- qPCR-R 0.4µl, cDNA 2µl, ddH2O 7.2µl, total system 20µl; reaction procedure is as follows: 95℃ pre-denaturation 5min, 95℃ denaturation 20s, 60℃ annealing 20s, 72℃ extension 20s, 40 cycles, using 2 -△△ct The data were analyzed by GraphPad Prism8 and plotted. Figure 3 shown.

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

[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 strain, ZmbZIP6 obtained in Example 3 OE1 strain, ZmbZIP6 OE2 The T0 generation plants of the strain were hybridized to obtain T1 generation plants. The T1 generation plants were used as research objects, and the number of days in their different growth stages was counted. The results are shown in Table 2 below.

[0050] Table 2 Statistics of reproductive period

[0051] As can be seen from Table 1, overexpression ZmbZIP6 The gene has almost no effect on the whole growth period of the plant, while knocking out ZmbZIP6 The gene causes a delay in the plant's full growth period.

[0052] Furthermore, ZmbZIP6 grown to the three-leaf and one-heart stage were placed in a plastic storage box with a length of 66 cm, a width of 42 cm, and a height of 40 cm. NT strain, ZmbZIP6 OE1 strain, ZmbZIP6 OE2ZmbZIP6 ko1 ZmbZIP6 ko2 ZmbZIP6 ZmbZIP6 ZmbZIP6 Figure 4 and 5 .

[0053] As can be seen from Figure 4 , after 15d of waterlogging stress treatment, ZmbZIP6 OE1 and ZmbZIP6 OE2 strains grow obviously better than ZmbZIP6 NT strain Figure 4 b, 4d, in two groups of treatment (normal growth and waterlogging stress treatment), the average plant height of ZmbZIP6 NT strain is 38.10 Figure 4 e) and 38.09 Figure 4 f) respectively, the average plant height of ZmbZIP6 OE1 strain is 41.20 Figure 4 e), and the average plant height of ZmbZIP6 OE2 strain is 41.53 Figure 4 f); the result shows that after overexpressing ZmbZIP6 gene, the waterlogging tolerance of plant is significantly improved.

[0054] As can be seen from Figure 5 , after 15d of waterlogging stress treatment, ZmbZIP6 ko1 and ZmbZIP6 ko2 strains grow obviously worse than ZmbZIP6 NT strain Figure 5 b, 5d, in two groups of treatment (normal growth and waterlogging stress treatment), the average plant height of ZmbZIP6 NT strain is 38.09 Figure 5 e) and 38.11 Figure 5 f) respectively, the average plant height of ZmbZIP6 ko1 strain is 27.62 Figure 5 e), and the average plant height of ZmbZIP6 ko2 strain is 27.66 Figure 5 f); the result shows that after knocking out ZmbZIP6 gene, the waterlogging tolerance of plant is significantly reduced.

[0055] In summary, the present application first finds that ZmbZIP6 gene and the protein coded by the gene are applied in regulating waterlogging tolerance of plant, which provides a theoretical basis for molecular breeding and germplasm improvement related to waterlogging tolerance.

[0056] The above embodiments only express the implementation of the present application, and the description is more specific and detailed, but it cannot be understood as a limitation on the scope of the patent. It should be noted that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of protection of the present application patent should be subject to the appended claims.

Claims

1. Application of ZmbZIP6 protein in regulating plant waterlogging tolerance, characterized in that: The ZmbZIP6 protein is selected from any one of the following: A1) a protein with an amino acid sequence as shown in SEQ ID NO. 1; A2) a protein having the same function as the amino acid sequence defined in A1) by substitution, deletion or addition of one or more amino acids; A3) A protein having the same function is obtained by ligating a tag to the N-terminus and / or C-terminus of A1) or A2).

2. Application of biomaterials in regulating waterlogging tolerance of plants, characterized in that: The biological material is selected from any one of the following: B1) a nucleic acid molecule encoding the ZmbZIP6 protein according to claim 1; B2) a nucleic acid molecule that inhibits or reduces the expression of the gene encoding the ZmbZIP6 protein according to claim 1; B3) a recombinant vector comprising the nucleic acid molecule described in B1) or B2); B4) a recombinant cell comprising the nucleic acid molecule of B1) or B2), or a recombinant cell comprising the recombinant vector of B3); B5) A transgenic plant comprising the nucleic acid molecule described in B1) or B2).

3. The use according to claim 2, characterized in that B1) The nucleic acid molecule is selected from any one 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) and encodes the ZmbZIP6 protein according to claim 1; C3) A nucleic acid molecule that has a sequence identity of more than 90% with the nucleic acid molecule defined in C1) or C2) and encodes the ZmbZIP6 protein according to claim 1.

4. The use according to claim 2, characterized in that B2) The nucleic acid molecule is an sgRNA, and the target sequence of the 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.

5. A method for cultivating a plant with reduced waterlogging tolerance, characterized in that: The method comprises reducing the content and / or activity of the ZmbZIP6 protein according to claim 1 in a target plant, thereby obtaining a plant having lower waterlogging tolerance than the target plant.

6. The method according to claim 5, characterized in that The reducing of the content and / or activity of the ZmbZIP6 protein according to claim 1 in the target plant is achieved by reducing the expression level of the gene encoding the ZmbZIP6 protein in the target plant.

7. The method according to claim 6, characterized in that The reducing the expression level of the gene encoding the ZmbZIP6 protein in the target plant is performed using the CRISPR / Cas9 system, and the CRISPR / Cas9 system includes the sgRNA according to claim 4.

8. A method for cultivating a plant with improved waterlogging tolerance, characterized in that: The method comprises increasing the content and / or activity of the ZmbZIP6 protein according to claim 1 in a target plant, thereby obtaining a plant having higher waterlogging tolerance than the target plant.

9. The method according to claim 8, characterized in that The increasing of the content and / or activity of the ZmbZIP6 protein according to claim 1 in the target plant is achieved by increasing the expression level of the gene encoding the ZmbZIP6 protein in the target plant.

10. The use according to any one of claims 1 to 4 or the method according to any one of claims 5 to 9, characterized in that: The plants include corn.

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