Application and method of wheat demethylase TaALKBH23 in regulation and control of drought stress resistance of plants

Abstract

The invention discloses application and a method of wheat demethylase TaALKBH23 in regulation and control of drought stress resistance of plants, a coding sequence of the demethylase TaALKBH23 is shown as SEQ ID NO: 1, and an amino acid sequence coded by the demethylase TaALKBH23 is shown as SEQ ID NO: 2. The invention provides demethylase TaALKBH23 and a function and application of a coding gene of the demethylase TaALKBH23 in regulation and control of plant drought resistance, and it is found through construction of a TaALKBH23 wheat genetic transformation material that the gene negatively regulates and controls wheat drought resistance. It is found that TaALKBH23 plays an important role in the process of regulating and controlling drought stress resistance of plants, and a new gene resource is provided for improving drought resistance of crops.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, and in particular to the application and method of wheat demethylase TaALKBH23 in regulating plant resistance to drought stress. Background Technology

[0002] Drought stress is a major limiting factor affecting crop growth, development, and yield. Drought causes a series of damages to the physiological, biochemical, and metabolic processes of plants, leading to slowed plant growth, cell damage, reduced crop yield and quality, and even plant death.

[0003] Wheat is mainly distributed in arid and semi-arid regions. As the main food crop in arid areas, sustained increases in wheat production are of great significance to ensuring food security. Discovering and utilizing drought-resistant genes to cultivate new drought-resistant crop varieties is the most direct and effective way to overcome the harm of drought.

[0004] N 6 -Methyladenosine (m 6 A) Modification is a dynamic and reversible process, initiated by methyltransferases (writers), reversed by demethylases (erasers), and bound by methyl recognition proteins (readers) to regulate RNA processing and metabolism. 6 RNA modification plays an important role in plant growth and development, morphogenesis, and stress response. RNA demethylation is achieved through the α-ketoglutarate-dependent dioxygenase (ALKB) homolog (ALKBH) protein.

[0005] Studies have shown that ALKBH10B has been reported as an m in Arabidopsis thaliana. 6 The demethylase alkbh10b mutant is sensitive to drought stress. Responses of demethylases to drought stress have also been reported in other species; for example, in tomato, the Slalkbh10b mutant shows increased tolerance to drought stress; in poplar, lines overexpressing PagALKBH10B are sensitive to drought stress due to decreased proline content; and in cotton, drought treatment enhances plant m 6 The A modification level was increased, and the alkbh10b mutant plants exhibited enhanced drought resistance. TaALKBH23, as a homolog of ALKBH10B in Arabidopsis, may also play an important role in regulating plant drought resistance.

[0006] However, no reports have been found on the role of demethylase genes in regulating drought stress in wheat.

[0007] Therefore, providing an application and method for wheat demethylase TaALKBH23 in regulating plant drought resistance is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0008] In view of this, the present invention provides the application and method of wheat demethylase TaALKBH23 in regulating plant resistance to drought stress.

[0009] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0010] The application of wheat demethylase TaALKBH23 in regulating plant resistance to drought stress, wherein the coding sequence of the demethylase TaALKBH23 is shown in SEQ ID NO: 1, and the encoded amino acid sequence is shown in SEQ ID NO: 2.

[0011] Preferably, the plant is a monocotyledonous plant.

[0012] Methods for regulating plant drought resistance using wheat demethylase TaALKBH23: This study utilizes genetic engineering techniques to obtain transgenic lines that overexpress and silence wheat demethylase TaALKBH23.

[0013] Preferably, the drought resistance is manifested as follows: compared with the wild type, under drought stress, the TaALKBH23 gene-silenced lines have less leaf wilting and yellowing, and the plant leaves have a lower relative water content, while the overexpressed lines have more severe leaf wilting and yellowing, and the plant leaves have a higher relative water content.

[0014] Preferably, the expression of the TaALKBH23 gene in wheat is regulated by overexpression and silencing of the TaALKBH23 gene.

[0015] A method for breeding transgenic plants with controlled drought resistance, wherein plants with drought resistance stronger or weaker than the target plant are obtained by regulating the expression of the TaALKBH23 gene of the target plant, the nucleotide sequence of which is shown in SEQ ID NO: 1.

[0016] Preferably, the target plant is wheat.

[0017] The present invention achieves the following technical effects compared to the prior art:

[0018] This invention, through the construction of TaALKBH23 wheat genetic transformation material, discovered that this gene negatively regulates wheat drought resistance; this invention also found that TaALKBH23 plays an important role in regulating plant resistance to drought stress, providing a new gene resource for improving crop drought resistance. Attached Figure Description

[0019] Figure 1 Genetic transformation map of transgenic TaALKBH23 overexpression;

[0020] Where, a is the map of the transgenic silencing vector; b is the map of the transgenic overexpression vector;

[0021] Figure 2 Molecular identification of T1 generation TaALKBH23 transgenic wheat lines;

[0022] Among them, (a) and (b) are molecular identification of overexpressed and silenced transgenic wheat, respectively. The Bar gene band size is 681 bp, M: Marker, and 1-12 are transgenic lines. (c) and (d) are qRT-PCR identification of overexpressed and silenced transgenic wheat.

[0023] Figure 3 Detection of methylation levels in transgenic wheat under normal conditions.

[0024] Figure 4 Phenotypic identification of TaALKBH23 transgenic wheat under drought stress.

[0025] (a) Phenotypic diagram of transgenic lines under natural drought (b) Relative water content of leaves of transgenic overexpression lines under natural drought (c) Relative water content of leaves of transgenic silenced lines under natural drought

[0026] Figure 5 Determination of physiological indicators in genetically modified wheat.

[0027] (a) (b) NBT staining of transgenic wheat under normal water conditions and natural drought stress (c) Chlorophyll content (d) Soluble sugar content.

[0028] Figure 6 Subcellular localization of TaALKBH23;

[0029] Wherein, ad represents the localization of the empty vector and the nuclear marker under green fluorescence field, red fluorescence field, bright field, and merge, respectively; eh represents the localization of the TaALKBH23 overexpression vector and the nuclear marker under green fluorescence field, red fluorescence field, bright field, and merge, respectively. Detailed Implementation

[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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 skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] This invention discloses the application of wheat demethylase TaALKBH23 in regulating plant resistance to drought stress. The coding sequence of demethylase TaALKBH23 is shown in SEQ ID NO: 1, and its encoded amino acid sequence is shown in SEQ ID NO: 2.

[0032] SEQ ID NO: 1:

[0033] ATGACGACAC CGGCGGCGGG GGCCCCGGGG TCACCGGTGG CGGCGCCGGA TCAGGTGGCG 60

[0034] GCCAGGGACG CCGTGATCGG GTGGTACCGC GGCGAGTTCG CGGCCGCCAA CGCCGTGATC 120

[0035] GACGCGCTGT GCGGCCACCT CGCGCAGATC GGCGGGGCCG ACTACGACGC CGTCTTCGCC 180

[0036] GCGCTCCACC GCCGCCGCCT CAACTGGTTC CCGTCCTCC ACATGCAGAA GTTCTACTCC 240

[0037] GTCGCCGACG TCGCCGCCGA GCTCCGTCGC GTCTCCGACG CCCGCGCCGC CGCGGCCGCC 300

[0038] GCCTTCTCCG AGGAGGAGGC CGCGTCCACG GTGATCCACG AGCCCATGGA CGAGCTCGTC 360

[0039] GTCACCGTCG CCGCGGAGCC GGACCCCGAG CCCGAGCACG AGCACGAGCC TATACCGGAG 420

[0040] GCGCAGCCCG ATGTCTCCGT CCACCCCGTC GTCGCCGTCC ACTCCGTCGA CGCGGACCGC 480

[0041] GAGCCGGAGG CGGACGGCGA AGATTCCTCG GGAGATTCGT CGGAGCGGAA GGCGGCCTCC 540

[0042] ACGGAAGACG ACGCCGCCCA CGACGGACCA GATAACACCG ACCAAGGATC TCAAGGCGGG 600

[0043] CACAGCCTAC CAGAGAGCTA CCCTATCTGC TCTGACCACG ACGAGTGCAT CGCCCGCCCC 660

[0044] GAGAGGATCA AGATCCAGAA AGGTTTCATG GCGAAGGAGT CCGTGAAGGG GCACATGGTT 720

[0045] AACGTTGTGA AAGGCCTGAA GATATGAA GACGTGTCCA WINTERGGA GCTCATGAAG 780

[0046] GTTGCTGATT TCATCAATGA AATTCGCCAG GCTGGTAGAA ATGGGGAGCT TTCAGGTGAA 840

[0047] ACCTTCATAT TCTTCAACAA GCAGATCAAA GGGAACAAGA GAGAGATCAT TCAGCTCGGT 900

[0048] GTCCCACTAT TTCAACATAC CACAGAGGAA ACCAATTGTC ATATAGAACC AATCCCAGTT 960

[0049] GTCCTGCAGG CTGTCATCGA CCACCTTGTT CTTTGGCGCT TAATACCAGA AAGCAGGAAA1020

[0050] CCAAACAGTG TCGTCATCAA TTTCTTCGAT GAGGACGAGC ACTCGCAGCC CTACTTCAAG1080

[0051] CCTCCCCACC TGGACAACCC CATTTCCACT CTCCTGCTGT CTGAGACCTC AATGGCATTT1140

[0052] GGAAGGTCAC TTGTCACTGA CAGCAATGGC AACTACAAGG GACCCCTCAC ACTCTCACTG1200

[0053] AAGCAAGGGT CACTTCTGGT GATGCGCGGG AACAGTGCGG ACATGGCGCG CCACGTTGTG1260

[0054] TGCCCGTCGT CCAACCGCCG CGTGAGCATC ACGTTCGTGA GGGTGAGGCC GTCGACGCCG1320

[0055] GTGGACCTCA GCCCGCTCCC GTCGCCCACC AAGGCCATGA CGCTCTGGCA GCCTCCCCCG1380

[0056] ACTGCGGCAA CGGCCGGCAT GCAGAAACCA CCCCATGGCA GCAACGGCGC CATCATTGGC1440

[0057] TACTGCCCGG CGCCACAGGC CATGCTCGCC CCTGCGTGGG GCATGGCCGT GCGCGCGGCG1500

[0058] CCTGTCATGA TGGTCGCCGC GCCGGCGAGG CCAATGGTGA TGGCGCCCTC CAGCAACATC1560

[0059] AACAAGAGGA TGGGGCGCGG CGGCACCGGC GTGTTCCTGC CGTGGACGGT TGGGCCCAAG1620

[0060] EXECUTE AGCACCTCCC CCCGCGCATC CAGAAGCGCC GGTTCTCGGC PRICEGTCG1680

[0061] CCCATAGAGT CGCAGGGCTG A

[0062] SEQ ID NO:2:

[0063] MTTPAAGAPG SPVAAPDQVA ARDAVIGWYR GEFAAANAVI DALCGHLAQI GGADYDAVFA 60

[0064] ALHRRRLNWF PVLHMQKFYS VADVAAELRR VSDARAAAAAAFSEEEEAAST VIHEPMDELV 120

[0065] VTVAAEPDPE PEHEHEPIPE AQPDVSVHPV VAVHSVDADR EPEADGEDSS GDSSERKAAS 180

[0066] TEDDAAHDGP DNTDQGSQGG HSLPESYPIC SDHDECIARP ERIKIQKGFM AKESVKGHMV 240

[0067] NVVKGLKIYE DVFTTMELMK VADFINEIRQ AGRNGELSGE TFIFFNKQIK GNKREIIQLG 300

[0068] VPLFQHTTEE TNCHIEPIPV VLQAVIDHLV LWRLIPESRK PNSVVINFFD EDEHSQPYFK 360

[0069] PPHLDNPIST LLLSETSMAF GRSLVTDSNG NYKGPLTLSL KQGSLLVMRG NSADMARHVV 420

[0070] CPSSNRRVSI TFVRVRPSTP VDLSPLPSPT KAMTLWQPPP TAATAGMQKP PHGSNGAIIG 480

[0071] YCPAPQAMLA PAWGMAVRAA PVMMVAAPAR PMVMAPSSNI NKRMGRGGTG VFLPWTVGPK 540

[0072] RYNKHLPPRI QKRRFSAMMS PIESQG 566

[0073] The plant is a monocotyledonous plant.

[0074] This invention also discloses a method for regulating plant drought resistance using wheat demethylase TaALKBH23, and uses genetic engineering techniques to obtain transgenic lines that overexpress and silence wheat demethylase TaALKBH23.

[0075] Drought resistance was demonstrated as follows: compared with the wild type, under drought stress, the TaALKBH23 gene-silenced lines showed less leaf wilting and yellowing, and the plant leaves had a lower relative water content, while the overexpressed lines showed more severe leaf wilting and yellowing, and the plant leaves had a higher relative water content.

[0076] The expression of the TaALKBH23 gene in wheat is regulated by overexpression and silencing of the TaALKBH23 gene.

[0077] This invention also discloses a method for cultivating transgenic plants with controlled drought resistance, which obtains plants with drought resistance stronger or weaker than the target plant by regulating the expression of the TaALKBH23 gene of the target plant, the nucleotide sequence of which is shown in SEQ ID NO: 1.

[0078] The target plant is wheat.

[0079] Experimental material: Chinese Spring, a variety of common wheat (Triticum aestivum L.). (Chinese Spring is a very important local wheat variety, widely used in wheat genetics research.)

[0080] Example 1: Full-length cloning and vector construction of the TaALKBH23 genome CDS

[0081] 1. Select whole, plump and uniform wheat seeds (Chinese Spring variety), sterilize with 75% alcohol for 10 min, rinse with sterile water, place in a 10 cm diameter petri dish with ceramsite, cover with a film and place in a 4 ℃ refrigerator for three days. Remove from the refrigerator, keep the filter paper moist, and culture at 22°C for two weeks to obtain wheat seedlings.

[0082] 2. Take wheat seedlings, flash-freeze them with liquid nitrogen, and store them at -80℃ for later use. Extract total RNA from wheat leaves using the Tiangen Total RNA Extraction Kit, and then reverse transcribe the RNA using the PrimeScript™ II 1st Strand cDNA Synthesis Kit (Takara) to obtain cDNA.

[0083] 3. Using wheat cDNA as a template, the following methods were employed:

[0084] 5'-ACGCATACGGGTAGGGAATTG-3'

[0085] PCR amplification was performed using a primer pair consisting of 5'-CCAAACTCGGCATGTATGTATGC-3' to obtain the PCR amplification product of the full-length CDS of TaALKBH23. The PCR product was detected by 1.0% agarose gel electrophoresis.

[0086] 4. The PCR reaction program was as follows: 95℃ pre-denaturation for 30 sec; 95℃ denaturation for 5 sec, 55℃ annealing for 30 sec, 72℃ extension for 1 min, for 35 cycles; 72℃ extension for 5 min.

[0087] 5. Design homologous arm primers to ligate the PCR amplification product into the overexpression vector, transform it into E. coli, and select single clones for sequencing. The homologous arm primers are as follows:

[0088] CUB-3×flag-TaALKBH23-F:

[0089] TGAAAAAGAGGGGGATTAACTAGTATGACGACACCGGCGGCGGGGGCC

[0090] CUB-3×flag-TaALKBH23-R:

[0091] ACCGCTACCACCGCTACCGAGCTCGCCCTGCGACTCTATGGGCGACAT

[0092] Sequencing results showed that the nucleotide sequence CDS of the PCR amplification product was consistent with the reference sequence. Figure 1 As shown in (a).

[0093] 6. After obtaining the target fragment through the SGN VIGS Tool website, the TaALKBH23 interference fragment was amplified. The interference fragment is shown in SEQ ID NO: 3.

[0094] SEQ ID NO: 3:

[0095] GAGAGGATCAAGATCCAGAAAGGTTTCATGGCGAAGGAGTCCGTGAAGGGGCACATGGTTAACGTTGTGAAAGGCCTGAAGATATATGAAGACGTGTTCACAACAATGGAGCTCATGAAGGTTGCTGATTTCATCAATGAAATTCGCCAG GCTGGTAGAAATGGGGAGCTTTCAGGTGAAACCTTCATATTCTTCAACAAGCAGATCAAAGGGAACAAGAGAGAGATCATTCAGCTCGGTGTCCCACTATTTCAACATACCACAGAGGAAACCAATTGTCATATAGAACCAATCCCAGTT

[0096] 7. Design homologous arm primers for the forward and reverse fragments and ligate them to the silencing vector.

[0097] Positive homologous arm primer:

[0098] CUB-RNAi-TaALKBH23-F-1:

[0099] GAAAAAGAGGGGGATTAGGATCCGAGAGGATCAAGATCCAGAAAGGTT

[0100] CUB-RNAi-TaALKBH23-R-1:

[0101] ACGCGAAGCGGGTAGATATCACTAGTAACTGGGATTGGTTCTATATGA

[0102] Reverse homologous arm primers:

[0103] CUB-RNAi-TaALKBH23-F-2:

[0104] ACTCGATCGAATTCCTGCAGCCCGGGAACTGGGATTGGTTCTATATGA

[0105] CUB-RNAi-TaALKBH23-R-2:

[0106] AACGATCGGGGAAAATTCGAGCTCGAGAGGATCAAGATCCAGAAAGGT

[0107] 8. Transform into E. coli and select single clones for sequencing.

[0108] Sequencing results showed that the nucleotide sequence CDS of the PCR amplification product was consistent with the reference sequence. Figure 1 As shown in (b).

[0109] Example 2: Establishment of transgenic lines of TaALKBH23

[0110] Wheat overexpression and silencing vectors were constructed and transformed into Agrobacterium. Identified positive single clones were then sent to the platform of the Key Laboratory of Arid Zone Crop Stress at Northwest A&F University for transformation. Vector-specific Bar gene primers were designed, and DNA was extracted from all wheat lines for PCR amplification. Positive lines were screened based on agarose gel electrophoresis results. RNA was extracted from positive plants and detected by qRT-PCR using a quantitative real-time PCR kit (including ROX) from Aike Rui Biotechnology, employing a QuantStudio 7 Flex amplification reaction. A two-step method was used with three technical replicates, and the PCR volume was 20 μL. The internal control gene was TaELF, and the reaction system and conditions were the same as above. Data were calculated using the 2-ΔΔCt method, and ANOVA and significance tests were performed using GraphPad Prism 9. * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001. Figure 2 The results showed that the expression levels of OE2 and OE46 were extremely significant in the six transgenic TaALKBH23 overexpression lines, and the expression levels of OE18, OE5, and OE9 were also significantly increased. Among the six transgenic TaALKBH23 silent lines, the expression levels of Ri28, Ri38, and Ri51 were the lowest. Based on the expression level detection results, the overexpression lines OE2, OE46, and OE18 were selected; the silent lines Ri28, Ri38, and Ri51 were used for multiple generations until they were stable for subsequent experiments.

[0111] Example 3: Detection of methylation levels in transgenic wheat under normal conditions

[0112] EpiQuik M was selected 6 A RNA Methylation Quantification Kit (M 6 A quantitative RNA methylation detection kit was used to detect methylation levels in wild-type, two overexpression lines, and two silenced lines under normal conditions. A single-point control method was employed. The entire experiment was conducted in the dark, and each sample was tested in duplicate.

[0113] The specific process is as follows:

[0114] (1) Calculate the number of 8 tubes required for the experiment, carefully remove them and put the remaining tubes back into the bag.

[0115] (2) Add 80 μL of BS binding solution to each well.

[0116] (3) Add 2 μL of NC negative control, 2 μL of 0.5 ng / μL PC positive control, and 200 ng of sample RNA to the corresponding wells. Gently shake the plate to mix the solutions and ensure the solution covers the bottom of the wells evenly.

[0117] (4) Cover with film and incubate at 37 ℃ for 90 min.

[0118] (5) Remove the BS solution from each well and wash three times with 150 μL 1×WB washing buffer.

[0119] (6) Add 50 μL of diluted CA capture antibody, cover and incubate at room temperature for 60 min.

[0120] (7) After removing the CA solution, wash three times with 150 μL of 1×WB.

[0121] (8) Add 50 μL of diluted DA detection antibody, cover and incubate at room temperature for 30 min.

[0122] (9) After removing the DA solution, wash four times with 150 μL of 1×WB.

[0123] (10) Add 50 μL of diluted ES enhancement solution, cover and incubate at room temperature for 30 min.

[0124] (11) Remove the ES solution and wash five times with 150 μL of 1×WB.

[0125] (12) Add 100 μL of DS developing solution to each well, incubate at room temperature in the dark for 1-10 min, and then monitor the color change of the sample.

[0126] (13) When the color of the positive control well turns medium blue, add 100 μL of SS blocking agent to prevent the enzymatic reaction.

[0127] (14) After adding SS, the color turns yellow, and then the absorbance value at 450 nm is read using an ELISA reader within 2-15 min.

[0128] Calculation method:

[0129]

[0130] Note: S is 200 ng, P is 1 ng.

[0131] Figure 3The results showed that, compared with the wild type, the methylation level of overexpressing plants was significantly reduced, while the methylation level of silenced plants was significantly increased. This indicates that the introduction of the TaALKBH23 gene does indeed affect the methylation level of the entire wheat plant.

[0132] Example 4: TaALKBH23 negatively regulates wheat drought resistance

[0133] Seedling drought resistance phenotype identification

[0134] Wheat plants that had grown normally for one month were divided into two groups for treatment. One group underwent natural drought treatment for 15 days, while the other group grew normally. The relative water content, soluble sugar, and chlorophyll content of leaves from the same part of the wheat plants under different treatments were measured. Simultaneously, NBT staining was performed on the same part of the wheat leaves from different treatments to detect superoxide anions, with three replicates for each indicator. Specific results are as follows: Figure 4 .

[0135] Figure 4 The results showed that after 15 days of natural drought treatment, the overexpressing lines exhibited more pronounced wilting and drying of leaves compared to the wild type, while the silenced lines showed better growth than the wild type. Five days after rehydration, a very clear phenotypic difference emerged between the wild type and the transgenic wheat: the wild type showed significantly better growth and survival than the overexpressing lines. The silenced lines showed the best overall growth and a higher survival rate. In summary, the soil culture phenotypic results indicate that the TaALKBH23 gene is a negatively regulated gene under drought stress.

[0136] Under normal conditions, NBT staining showed no difference among the lines. However, after drought stress treatment, the leaves of the overexpressing lines accumulated a large amount of blue precipitate, followed by the wild-type lines, with the silent lines accumulating the least, indicating that the overexpressing lines accumulated more superoxide anions. Chlorophyll content also showed no significant difference under normal conditions, but after drought stress treatment, compared with the wild type, the silent lines had higher chlorophyll content, while the overexpressing lines had lower content. Drought stress has a significant impact on chlorophyll synthesis in wheat leaves. Soluble sugars are important regulators of plant growth, development, and gene expression. As small molecule organic compounds, they are important solutes involved in regulating osmotic pressure within plant cells. Under osmotic stress, large amounts of soluble sugars accumulate to reduce osmotic potential, thereby resisting stress damage. Soluble sugar content showed no significant difference under normal conditions. Under drought treatment, the soluble sugar content, from highest to lowest, was in the silent lines, wild-type, and overexpressing lines.

[0137] These physiological indicators further demonstrate that the silent transgenic lines under drought stress exhibit better drought resistance in both morphology and physiology.

[0138] Example 5: Subcellular localization of the TaALKBH23-GFP fusion protein

[0139] Using the correctly sequenced recombinant cloning plasmid obtained in Example 1 as a template, 5'-GCAAGTTCTTCACTGTTGATACATATGACGACACCGGCGGCG-3' and 5'-CTACCACCGCTACCGTCGACGCCCTGCGACTCTATGGGCGACATC-3' were used as primers for PCR product amplification. The CDS sequence of the target gene was ligated into the expression vector 35S::-GFP to obtain the recombinant vector 35S::TaALKBH23-GFP, which was then transformed into Agrobacterium GV3101. Transient expression was performed in tobacco leaves with the empty vector 35S::GFP as a control, and the results were observed under a laser confocal microscope.

[0140] Figure 6 The results showed that the empty vector could be observed in both the nucleus and cell membrane of tobacco leaves, while the 35s::TaALKBH23-GFP was only observed in the nucleus and co-localized with the nuclear localization maker in the merge, confirming that TaALKBH23 is localized in the nucleus, indicating that the gene may be expressed in the nucleus and play an important role.

[0141] The above description is merely a preferred embodiment of the present invention and does not constitute any limitation on the technical scope of the present invention. Therefore, any minor modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention shall still fall within the scope of the technical solution of the present invention.

Claims

1. Application of wheat demethylase TaALKBH23 in regulating plant drought stress resistance, characterized in that, The demethylase TaALKBH23 coding sequence is shown as SEQ ID NO: 1, and the encoded amino acid sequence is shown as SEQ ID NO:

2.

2. The use of wheat demethylase TaALKBH23 in regulating plant drought stress resistance according to claim 1, characterized in that, The plant is a monocotyledon.

3. A method for regulating drought stress resistance in plants using wheat demethylase TaALKBH23, characterized in that, By means of genetic engineering, overexpression and silencing transgenic lines of wheat demethylase TaALKBH23 are obtained.

4. The method of claim 3, wherein the wheat demethylase TaALKBH23 is used for regulating drought stress resistance in plants. The drought resistance is shown as follows: compared with the wild type, under drought stress, the leaf wilting and yellow flower of the TaALKBH23 gene silencing line is lighter, the relative water content of the plant leaf is lower, while the leaf wilting and yellow flower of the overexpression line is heavier, and the relative water content of the plant leaf is higher.

5. The method of claim 3, wherein the wheat demethylase TaALKBH23 is used for regulating drought stress resistance in plants. The expression mode of the TaALKBH23 gene in wheat is overexpression and silencing of the TaALKBH23 gene.

6. A method of breeding a transgenic plant that modulates drought resistance, comprising, By regulating the expression of the TaALKBH23 gene in the target plant, a plant with stronger / weaker drought resistance than the target plant is obtained, and the nucleotide sequence of the gene is shown as SEQ ID NO:

1.

7. The method of breeding a transgenic plant having improved drought resistance according to claim 6, wherein, The target plant is wheat.

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