Application of wheat disease resistance factor TaCDPK28 in the control of stripe rust

Abstract

The present invention belongs to the field of genetic engineering technology and relates to the application of wheat disease resistance factor TaCDPK28 in the prevention and treatment of stripe rust. The present invention provides a TaCDPK28 protein, which has an amino acid sequence as shown in SEQ ID NO: 1 and is encoded by a nucleotide sequence as shown in SEQ ID NO: 2. It has been verified that the TaCDPK28 protein provided by the present invention has an amino acid sequence as shown in SEQ ID NO: 1 and is encoded by a nucleotide sequence as shown in SEQ ID NO: 2. TaCDPK28 When the gene was overexpressed, the severity of stripe rust in wheat plants was weaker than that in control plants, and the production of summer spores on wheat leaves was significantly reduced, providing genetic resources and technical ideas for breeding and improving wheat stripe rust resistance.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering and relates to the application of wheat disease resistance factor TaCDPK28 in the prevention and treatment of stripe rust. Background Art

[0002] Wheat Stripe Rust is a disease caused by Puccinia striae (Puccinia striae, Puccinia striiformis f. sp. Tritici , PST ) is an airborne fungal disease caused by the stripe rust of wheat, which can harm the entire growth period of wheat and seriously affect the yield and quality of wheat. The prevention and control of wheat stripe rust is mainly through the use of chemical pesticides and fungicides and the planting of disease-resistant varieties. However, the extreme biological selection pressure caused by the use of chemical pesticides can easily produce drug-resistant strains, and large-scale use can also cause environmental problems. This has led to an increase in the use of fungicides with the emergence of drug-resistant strains, but it has also increased the environmental carrying capacity and caused environmental damage. At the same time, as the prevention and control effect weakens, the cost of wheat cultivation has become increasingly high. Therefore, the promotion and planting of disease-resistant varieties of wheat stripe rust that are relatively safe for the environment is a relatively economical and effective prevention and control measure. However, the use of disease-resistant varieties also faces the problem of loss of host resistance, so there is an urgent need to explore more disease-resistant genes. Summary of the invention

[0003] The present invention screens TaCDPK28 protein and TaCDPK28 Genes, by functional analysis of the proteins and genes, reveal their disease resistance mechanism in the interaction between wheat and stripe rust, and provide solutions for the cultivation of stripe rust-resistant materials. To achieve this technical purpose, the present invention adopts the following technical solutions.

[0004] The present invention provides a wheat disease resistance-related TaCDPK28 protein and TaCDPK28 Gene, derived from wheat variety Shuiyuan 11 ( Triticum aestivum ). The TaCDPK28 protein has an amino acid sequence as shown in SEQ ID NO: 1, TaCDPK28 The gene comprises a nucleotide sequence encoding the TaCDPK28 protein.

[0005] Furthermore, overexpression of TaCDPK28 Genes improve wheat resistance to stripe rust.

[0006] For example, the TaCDPK28 The CDS sequence in the gene is shown in SEQ ID NO:2.

[0007] In another aspect, the present invention relates to an expression vector comprising the TaCDPK28 Gene.

[0008] In another aspect, the present invention relates to a method for breeding a wheat variety resistant to stripe rust, wherein the expression vector is transferred into wheat to overexpress the TaCDPK28 Gene.

[0009] Specifically, the present invention relates to a method for breeding wheat varieties resistant to stripe rust, comprising: overexpressing TaCDPK28 Gene improves wheat resistance to stripe rust TaCDPK28 The gene comprises a nucleotide sequence encoding a TaCDPK28 protein, and the TaCDPK28 protein has an amino acid sequence as shown in SEQ ID NO:1.

[0010] Furthermore, in the method for breeding wheat stripe rust resistant varieties provided by the present invention, the TaCDPK28 The CDS sequence of the gene is shown in SEQ ID NO:2.

[0011] Furthermore, in the method for breeding wheat stripe rust resistant varieties provided by the present invention, the TaCDPK28 The gene vector was connected by TaCDPK28-CUB and the blank vector after restriction digestion;

[0012] The amplification primers of TaCDPK28-CUB are TaCDPK28-CUBF shown in SEQ ID NO: 3 and TaCDPK28-CUBR shown in SEQ ID NO: 4.

[0013] On the other hand, the present invention relates to the use of TaCDPK28 protein and the gene encoding the same in the cultivation of stripe rust resistant materials. TaCDPK28 The gene includes a nucleotide sequence encoding TaCDPK28 protein, overexpressing the TaCDPK28 The gene improves the resistance of the host plant to stripe rust, and the TaCDPK28 protein has the amino acid sequence shown in SEQ ID NO:1.

[0014] Furthermore, in the application provided by the present invention, the nucleotide sequence encoding the TaCDPK28 protein is overexpressed and / or the concentration of the TaCDPK28 protein is increased to improve the resistance of the host plant to stripe rust.

[0015] Further, in the application provided by the present invention, the TaCDPK28 The CDS sequence of the gene is shown in SEQ ID NO:2.

[0016] A person skilled in the art can easily mutate the nucleotide sequence encoding TaCDPK28 of the present invention by using known methods, such as directed evolution and point mutation. Those artificially modified nucleotides having 75% or higher identity with the nucleotide sequence of TaCDPK28 isolated from the present invention are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention as long as they encode TaCDPK28 and have the same function.

[0017] The term "identity" as used herein refers to sequence similarity to a natural nucleic acid sequence. "Identity" includes nucleotide sequences that have 75% or more, or 85% or more, or 90% or more, or 95% or more identity to the nucleotide sequence of the protein composed of the amino acid sequence shown in the coding sequence 2 of the present invention. Identity can be evaluated with the naked eye or computer software. Using computer software, the identity between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the identity between related sequences.

[0018] The above-mentioned 75% or more identity may be 80%, 85%, 90% or 95% or more identity.

[0019] The expression cassette (TaCDPK28 gene expression cassette) containing a nucleic acid molecule encoding TaCDPK28 as described above refers to a DNA capable of expressing TaCDPK28 in a host cell, which may include not only a promoter for initiating TaCDPK28 transcription, but also a terminator for terminating TaCDPK28 transcription. Further, the expression cassette may also include an enhancer sequence. Promoters that can be used in the present invention include, but are not limited to, constitutive promoters; tissue, organ and development-specific promoters and inducible promoters. Suitable transcription terminators include, but are not limited to, Agrobacterium nopaline synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator, and nopaline and octopine synthase terminators.

[0020] The recombinant vector containing the TaCDPK28 gene expression cassette can be constructed using an existing expression vector. The plant expression vector includes a binary Agrobacterium vector and a vector that can be used for plant microprojectile bombardment, etc. Such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1305, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA Company), etc. The plant expression vector can also contain the 3' non-translated region of the foreign gene, that is, a polyadenylic acid signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylic acid signal can guide polyadenylic acid to be added to the 3' end of the mRNA precursor, such as the Agrobacterium crown gall induction (Ti) plasmid gene (such as the nopaline synthase gene Nos), the 3' non-translated region of the plant gene (such as the soybean storage protein gene) has a similar function. When using the gene construction plant expression vector of the present invention, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be identical to the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The source of the translation control signal and the start codon is extensive and can be natural or synthetic. The translation initiation region can come from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS gene, luciferase gene, etc.), antibiotic marker genes (such as nptII gene that confers resistance to kanamycin and related antibiotics, bar gene that confers resistance to the herbicide phosphinothricin, hph gene that confers resistance to the antibiotic hygromycin, dhfr gene that confers resistance to methotrexate, EPSPS gene that confers resistance to glyphosate), or chemical resistance marker genes (such as herbicide resistance genes), mannose-6-phosphate isomerase gene that provides the ability to metabolize mannose. Considering the safety of transgenic plants, no selective marker genes can be added, and transformed plants can be directly screened by adversity.

[0021] The above-mentioned vector or expression vector can be a plasmid, cosmid, phage or virus vector.

[0022] The microorganisms may be yeast, bacteria, algae or fungi, such as Agrobacterium.

[0023] The above transgenic plant cell lines do not include propagation materials.

[0024] The above disease resistance is resistance to stripe rust.

[0025] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0026] The present invention provides the application of wheat disease resistance factor TaCDPK28 in the prevention and treatment of stripe rust. The experiment of the present invention proves that the resistance of TaCDPK28 overexpression plants obtained by transgenic to stripe rust is higher than that of control plants, indicating that TaCDPK28 protein has the function of stripe rust resistance. The protein and gene provided by the present invention provide a basis for artificial control of stripe rust, and will play an important role in cultivating plants with enhanced stripe rust resistance. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solution of the present invention, the drawings involved in the description of the embodiments will be briefly introduced below. Obviously, the drawings in the description are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0028] Picture 1 for TaCDPK28 Histogram of gene expression characteristics in wheat and stripe rust interaction combinations. Picture 1 A in the figure is the time and TaCDPK28 Relationship diagram of relative gene expression; Picture 1 B in the figure is the time and TaCDPK28 Relative gene expression levels.

[0029] Picture 2 This is the localization result of TaCDPK28 protein in wheat protoplasts.

[0030] Picture 3 for TaCDPK28 Positive identification of the T1 generation of gene overexpressing transgenic plants and phenotypic analysis after inoculation with stripe rust. Picture 3 A in TaCDPK28 Picture of summer spore pile after inoculation of CYR34 with gene overexpression vector; Picture 3 B in the figure is the result of Western Blot test; Picture 3 The C in it is the PCR test result; Picture 3 D in the figure is a statistical bar graph of summer spore numbers; Fielder is a wild-type control plant inoculated with CYR34, and OE9, OE11, and OE24 are positive in both Western Blot and PCR tests. TaCDPK28 Gene overexpression transgenic plants. DETAILED DESCRIPTION

[0031] The technical scheme of the present invention is described below in conjunction with the examples, but the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are conventional methods unless otherwise specified; the reagents and materials are available on the market unless otherwise specified, and the % in the following examples are all mass percentages unless otherwise specified.

[0032] Example 1

[0033] This example provides the acquisition of TaCDPK28 protein and its encoding gene.

[0034] Wheat seedlings from water source 11 that had grown normally for 7 days were taken, quickly frozen with liquid nitrogen, and stored at -80°C for later use.

[0035] Total RNA from wheat leaves was extracted using the Trizol method (TianGen), and reverse transcriptase XL (AMV) was used for first-strand cDNA synthesis. cDNA was synthesized using the SMART method, and PCR products were detected by 1.0% agarose gel electrophoresis. The amplification primers were:

[0036] TaCDPK28-F: 5'-ATGCAGCCGGACCCTAAA-3';

[0037] TaCDPK28-R: 5'-TTAGGTGGTTCCACCGGG-3'.

[0038] A PCR product of 1563 bp was obtained.

[0039] After sequencing, the PCR product has the nucleotides shown in SEQ ID NO: 2 in the sequence list, and the gene with the nucleotides is named TaCDPK28 The amino acid sequence of the protein encoded by the gene is shown in SEQ ID NO: 1 in the sequence table, and the protein is named TaCDPK28 protein.

[0040] Example 2

[0041] This example provides RT-PCR detection TaCDPK28 Gene expression induced by stripe rust.

[0042] Wheat was inoculated with stripe rust at the one-leaf-one-heart stage. The inoculation method is recorded in the literature "Kang Zhensheng, Li Zhenqi. Discovery of a new pathogenic fungus strain of Lovelin 10 at room temperature [J]. Journal of Northwest Agriculture and Forestry University (Natural Science Edition), 1984 (04): 18-28." Wheat water source 11 leaves were inoculated with non-compatible combination race CYR23 and compatible combination race CYR31, and sterile water was used as a control.

[0043] Samples were taken at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 96 h, and 120 h after inoculation, and the sampling time of the control was consistent with that of the treatment. When sampling, fresh leaves were cut, wrapped in tin platinum paper, and quickly frozen in liquid nitrogen, and then stored at -80˚C for later use. The total RNA of wheat leaves was extracted by Trizol method (TianGen), and reverse transcriptase XL (AMV) was used for the first-strand cDNA synthesis. The cDNA was synthesized by SMART method.

[0044] Specific quantitative PCR primers were designed based on the sequences of wheat TaCDPK28 and elongation factor gene TaEF-1α.

[0045] The RT-PCR primer sequences are:

[0046] QTaCDPK28-F: 5'-TGTGTGTGGCCTCCGTTTTG-3'

[0047] QTaCDPK28-R: 5'-ACTAGGCCATGGTTCAGTTTCC-3'.

[0048] QTaEF-F: 5'-TGGTGTCATCAAGCCTGGTATGGT-3'

[0049] QTaEF-R: 5'-ACTCATGGTGCATCTCAACGGACT-3'

[0050] Before using the quantitative PCR primers, the specificity and amplification efficiency of their amplification products should be tested (≥90%). TaEF-1α was used as an internal reference gene in the real-time PCR analysis. AceQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) and Bio-Rad CFX Manager quantitative PCR instrument (Bio-rad, Hercules, California) were used to perform real-time quantitative PCR amplification using cDNA from each treatment sampling point as template according to the instructions. Each reaction was repeated at least 3 times, and the Ct value of each repeat and its average and standard deviation were generated by the quantitative PCR instrument by manually adjusting the baseline. Each reaction was repeated 3 times, and the Ct value was averaged. The experimental data were analyzed by the Delta Delta Ct method to determine the relative expression of the gene.

[0051] The results of qRT-PCR are as follows Picture 1As shown in the figure, the expression patterns of TaCDPK28 at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 96 h, and 120 h after wheat water source 11 was inoculated with the non-compatible race CYR23 of stripe rust and the compatible race CYR31 of stripe rust, respectively. TaCDPK28 showed an up-regulated expression trend in the early stage of stripe rust infection (24-48 h) in both the non-compatible and compatible combinations, with the highest expression levels of 6.9 times and 4.9 times, respectively. "*" indicates significance at the p<0.05 level compared with 0 h.

[0052] The above results show that TaCDPK28 The gene expression is induced by stripe rust.

[0053] Example 3

[0054] This example provides subcellular localization analysis of TaCDPK28 protein.

[0055] The above-mentioned PCR product TaCDPK28 fragment was used as a template and PCR amplification was performed using primers with BamHI restriction sites. The amplified product was digested with BamHI and connected with the 16318hGFP (green fluorescent protein) vector that was similarly digested to obtain the recombinant vector GFP-TaCDPK28 to express the fusion protein.

[0056] The amplification primers used for the construction of TaCDPK28 subcellular localization vector are:

[0057] TaCDPK28-GFP-F:5'-TATCTCTAGAGGATCCATGCAGCCGGACCCTAAA-3'

[0058] TaCDPK28-GFP-R:5'-TGCTCACCATGGATCCGGTGGTTCCACCGGG-3'.

[0059] Wheat varieties planted in soil culture room Water source 11. In case of good growth, take leaves before flowering to prepare protoplasts. Cut the leaves with good growth in the middle and cut them into 0.5-1 mm wide leaf strips with a blade. Put the cut leaf strips into the pre-prepared cellulose enzymatic hydrolysate shown in Table 1 (about 10-20 leaves are required for every 5-10 mL enzymatic hydrolysate).

[0060] Table 1: Cellulose enzymatic hydrolysate formula

[0061]

[0062] Use forceps to completely immerse the leaves in the enzymatic solution. Use a vacuum pump to evacuate the leaves in the dark (wrapped in foil) for 30 minutes. Continue the enzymatic solution in the dark for at least 3 hours without shaking at room temperature (shake at 50 rpm at 28°C for wheat). When the enzymatic solution turns green, gently shake the culture dish to release the protoplasts. Examine the protoplasts in the solution under a microscope. The size of wheat mesophyll protoplasts is about 30-50 μm. Dilute the enzyme solution containing protoplasts with an equal amount of W5 solution before filtering out the undissolved leaves. Wet a 35-75 μm nylon membrane or a 60-100 mesh sieve with W5 solution, and then use it to filter the enzymatic solution containing protoplasts. Centrifuge in a 30mL round-bottom centrifuge tube at 100g, 1-2min, 4°C to precipitate the protoplasts and remove as much supernatant as possible. Then gently resuspend the protoplasts in 10mL of ice-cold W5 solution. Let the protoplasts stand on ice for 30 minutes.

[0063] The following operations were performed at room temperature (23°C): centrifugation at 100 g for 8-10 min to precipitate the protoplasts. The W5 solution was removed as much as possible without touching the protoplast pellet; then the protoplasts were resuspended with an appropriate amount of MMG solution (1 M) to a final concentration of 2 × 10 5 / mL; add 10 μL or 20 μL DNA (10-20 micrograms of about 5-10 kb recombinant vector 16318GFP-TaCDPK28) to a 2 mL EP tube; add 100 μL protoplasts (2×10 4 ), gently mix; add 110 μL PEG solution, gently tap the centrifuge tube to mix completely (about 6-10 samples can be transformed each time); induce the transformation mixture for 20-30 minutes; dilute the transformation mixture with 400-440 μL W5 solution at room temperature, and then gently shake the centrifuge tube upside down to mix it well to terminate the transformation reaction; centrifuge at 100g for 2 minutes at room temperature, and then remove the supernatant; add 1mL W5 solution to suspend and wash once, centrifuge at 100g for 2 minutes to remove the supernatant; gently resuspend the protoplasts in a multi-well tissue culture dish with 1mL WI solution.

[0064] Protoplasts were induced at room temperature (20-25°C) for more than 18 hours, and the 16318GFP vector was transferred as a control. The expression of GFP tag was then observed under a laser confocal microscope.

[0065] After 18 hours of dark culture, the protoplasts were pressed into slices, and then the GFP (green fluorescent protein) fluorescence was observed under a laser scanning confocal microscope (Bio-Rad MicroRadiance) (LSMC), and scanning and photography were performed. The working parameters of LSCM were: Ex = 488nm, Em = 525 ± 15nm, Power = 10%, Zoom 7, medium-speed scanning, Frame 512 × 512. The software used was TIME-COURSE and PHOTOSHOP 5.0.

[0066] Results Picture 2 The upper one is the control of protoplasts transformed with the recombinant vector 16318GFP empty vector; the lower one is the localization map of TaCDPK28 in protoplasts transformed with the recombinant vector TaCDPK28-GFP. Picture 2 It can be seen that TaCDPK28 protein is distributed in the cytoplasm.

[0067] Example 4

[0068] This embodiment provides TaCDPK28 Application of gene enhanced expression in regulating plant resistance to stripe rust.

[0069] 1. TaCDPK28 Construction of gene CUB vector system

[0070] (1) Obtaining the overexpression sequence TaCDPK28-CUB

[0071] The amplified DNA in Example 1 TaCDPK28 The gene fragment (SEQ ID NO: 2) was used as a template, and primers TaCDPK28-CUBF and TaCDPK28-CUBR were used for PCR amplification to obtain a PCR amplification product of 1641 bp, which was named as the overexpression sequence TaCDPK28-CUB.

[0072] The nucleotide sequences of the above primer pair TaCDPK28-CUBF and TaCDPK28-CUBR are as follows:

[0073] TaCDPK28-CUBF (SEQ ID NO: 3): 5'-caggtcgactctagaggatccATGCAGCCGGACCCTAAA-3';

[0074] TaCDPK28-CUBR (SEQ ID NO: 4): 5'-gagctcggtacccggggatccTCACTTATCATCATCATCCTTATAATCTCCCTTATCATCATCCTTATAATCTCCCTTATCATCATCATCCTTATAATCGGTGGTTCCACCGGG-3'.

[0075] (2) Construction of CUB overexpression vector

[0076] The CUB overexpression vector was digested with BamH1, and then the overexpression sequence TaCDPK28-CUB obtained in the above step (1) was connected with the digested CUB overexpression vector to obtain the recombinant vector Vector-TaCDPK28-CUB.

[0077] 2. TaCDPK28 Obtaining gene overexpression transgenic plants

[0078] The recombinant vector Vector-TaCDPK28-CUB constructed in the above steps was transformed into the Agrobacterium strain EHA105, and the Agrobacterium culture plate was obtained after screening with kanamycin and rifampicin. Wheat transgenic operation was carried out through the wheat transgenic platform of the National Key Laboratory of Crop Stress and High Efficiency to obtain TaCDPK28 overexpressing transgenic plants.

[0079] T1 generation TaCDPK28 overexpressing transgenic plants and controls were planted and inoculated with stripe rust race CYR34 after their second leaves unfolded. The disease development was observed 14 days after inoculation.

[0080] The reaction type was identified according to the nine-type classification standard of Hovmøller (Hovmøller MS, Rodriguez-Algaba J., Thach T., Sørensen CK (2017) Race Typing of Puccinia striiformis on Wheat. In: Periyannan S. (eds) Wheat Rust Diseases. Methods in Molecular Biology, vol1659. Humana Press, New York, NY).

[0081] The results are as follows Picture 3 As shown in A, Fielder: wild-type control plant inoculated with CYR34; OE9, OE11, OE24: positive by Western Blot and PCR TaCDPK28Gene overexpression transgenic plants; under stripe rust infection conditions, visible uredia were observed on leaves of all treatments. TaCDPK28 The severity of stripe rust in gene overexpression transgenic plants was weaker than that in control plants.

[0082] Statistical analysis of leaf spore numbers showed that every 0.5 cm 2 The number of spore piles decreased significantly ( Picture 3 D in the figure). And detected by Western Blot ( Picture 3 B) and PCR detection ( Picture 3 C) Identification of transgenic plants overexpressing TaCDPK28 protein.

[0083] The results showed that wheat leaves overexpressing TaCDPK28 protein had improved resistance to stripe rust physiological race CYR34. This proved that TaCDPK28 is an important gene related to stripe rust resistance, and increasing the expression of this gene can improve plant resistance to stripe rust.

[0084] The above-described embodiments are part of the embodiments of the present invention, but not all of them. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but rather represents selected embodiments of the present invention. All other embodiments obtained without creative work and related deductions and substitutions made by ordinary technicians in the field under the conditions of the concept of the present invention belong to the scope of protection of the present invention.

Claims

1. A method for breeding wheat varieties resistant to stripe rust, characterized in that: include: Overexpression TaCDPK28 Gene improves wheat resistance to stripe rust TaCDPK28 The gene includes a nucleotide sequence encoding a TaCDPK28 protein, and the amino acid sequence of the TaCDPK28 protein is shown in SEQ ID NO:

1.

2. The method for breeding wheat stripe rust resistant varieties according to claim 1, characterized in that: Said TaCDPK28 The CDS sequence of the gene is shown in SEQ ID NO:

2.

3. The method for breeding wheat stripe rust resistant varieties according to claim 2, characterized in that: For overexpression of TaCDPK28 The gene vector was connected by TaCDPK28-CUB and the blank vector after restriction digestion; The TaCDPK28-CUB was obtained by PCR amplification using SEQ ID NO: 2 as a template and a primer pair; The primer pair is TaCDPK28-CUBF shown in SEQ ID NO: 3 and TaCDPK28-CUBR shown in SEQ ID NO:

4.

4. Application of TaCDPK28 protein in the cultivation of wheat stripe rust resistant materials, characterized in that: Overexpression of the gene encoding the TaCDPK28 protein improves wheat resistance to stripe rust. The amino acid sequence of the TaCDPK28 protein is shown in SEQ ID NO:

1.

5. TaCDPK28 The application of the gene in the cultivation of wheat stripe rust resistant materials is characterized in that: Said TaCDPK28 The gene includes a nucleotide sequence encoding TaCDPK28 protein, overexpressing the TaCDPK28 The gene improves wheat resistance to stripe rust. The amino acid sequence of the TaCDPK28 protein is shown in SEQ ID NO:

1.

6. The use according to claim 5, characterized in that: Overexpressing the nucleotide sequence encoding the TaCDPK28 protein and / or increasing the concentration of the TaCDPK28 protein can improve the resistance of wheat to stripe rust.

7. The use according to claim 5, characterized in that: Said TaCDPK28 The CDS sequence of the gene is shown in SEQ ID NO:2.

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