Wheat disease resistance related protein ta cngc-ps1 and gene and application thereof

Abstract

The application discloses a wheat disease-resistant protein TaCNGC-Ps1 and a gene and application thereof, and belongs to the technical field of genetic engineering. TaCNGC-Ps1 The application discloses a wheat disease-resistant protein TaCNGC-Ps1 and a gene and application thereof, and belongs to the technical field of genetic engineering. The application discloses a wheat disease-resistant protein TaCNGC-Ps1 and a gene and application thereof, and belongs to the technical field of genetic engineering.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of genetic engineering, and relates to a wheat disease-resistant related protein TaCNGC-Ps1 and a gene and application thereof. BACKGROUND

[0002] Wheat scab ( Fusarium head blight, FHB) is a kind of fungal disease that seriously damages wheat production. Chemical control is the main means for field control of wheat scab at present. However, long-term use of a single pesticide leads to resistance of the pathogen to the fungicide. Breeding of disease-resistant varieties is the most economical, effective and environmentally friendly strategy for preventing scab. Although breeders have improved the resistance of wheat to scab through traditional breeding methods, there is still a lack of stable scab-resistant varieties. Therefore, mining of scab-resistant genes and breeding of stable disease-resistant varieties are still the main tasks of wheat scab-resistant breeding at the present stage.

[0003] Wheat powdery mildew caused by the obligate biotrophic fungus Blumeria graminis f. sp. Blumeria graminis f.sp. tritici (Bgt) is an important disease in wheat production. Blumeria infection affects the growth and development of wheat seedlings, and reduces the yield of wheat at the adult stage. The most economical and effective measure to prevent and control wheat powdery mildew is to use disease-resistant varieties. However, due to the single resistance source and the rapid variation of the physiological race of the pathogen, disease-resistant varieties have successively “lost” resistance, and the area of wheat powdery mildew has increased year by year. The yield loss caused by powdery mildew is 5%~8% per year. Therefore, in-depth study of genes involved in wheat resistance to powdery mildew and mining of new gene resources will contribute to the development and application of future wheat disease-resistant molecular breeding.

[0004] In nature, plants have evolved sophisticated defense mechanisms to resist the infection of pathogenic microorganisms. These defense mechanisms include the host plant defense response (PAMP-triggered immunity, PTI) and the effector protein-induced defense response (Effector-triggered immunity, ETI). ETI is usually accompanied by the occurrence of hypersensitive response (HR). HR is the most common form of disease resistance in plants, which is manifested as local necrosis at the infection site and limits the growth of pathogenic bacteria. Because MAMPs (microbe-associated molecular patterns), PAMPs (pathogen-associated molecular patterns) or DAMPs (damage-associated molecular patterns) are very conservative, PTI is easily recognized by most pathogenic bacteria, and effector proteins have high specificity, so that ETI differs between different species or different physiological races. Although the activation of effector protein-induced defense response and host plant defense response is different, both can induce a series of plant immune responses, including ion flow on the lipid membrane, increase of intracellular calcium ion concentration, formation of reactive oxygen species (ROS), activation of MAPK (Mitogen-activated protein kinase) signaling pathway, etc. Subsequent reactions include secretion of antibacterial proteins, cell wall lignification, etc.

[0005] Therefore, mining wheat disease resistance related genes, understanding the response and signal transduction mechanism of wheat under the stress of Fusarium graminearum and powdery mildew, and improving the disease resistance of wheat have become important means for sustainable control of wheat scab and powdery mildew. SUMMARY

[0006] The purpose of the present application is to provide a wheat disease resistance related protein to improve the resistance of wheat to scab and powdery mildew.

[0007] In a first aspect, the present application provides a method for breeding a wheat scab-resistant variety, overexpressing a wheat disease resistance related protein gene TaCNGC-Ps1 ;

[0008] The nucleotide sequence of the wheat disease resistance related protein gene TaCNGC-Ps1 is shown as SEQ ID NO. 2.

[0009] Further, in the method for breeding a wheat scab-resistant variety provided by the present application, a recombinant expression vector is constructed, and the recombinant expression vector is transformed into wheat; the recombinant expression vector comprises the wheat disease resistance related protein gene TaCNGC-Ps1 .

[0010] In a second aspect, the present application provides a method for breeding a wheat variety resistant to powdery mildew, overexpressing a wheat disease resistance related protein gene TaCNGC-Ps1 ;

[0011] The nucleotide sequence of the wheat disease resistance related protein gene TaCNGC-Ps1 is shown as SEQ ID NO. 2.

[0012] Further, in the method for breeding a wheat variety resistant to powdery mildew provided by the present application, the method comprises constructing a recombinant expression vector, and transforming the recombinant expression vector into wheat; the recombinant expression vector comprises the wheat disease resistance related protein gene TaCNGC-Ps1 .

[0013] In a third aspect, the present application provides a wheat disease resistance related protein gene TaCNGC-Ps1 and an application of a protein encoded by the wheat disease resistance related protein gene in breeding wheat resistant to scab and / or powdery mildew, the nucleotide sequence of the wheat disease resistance related protein gene TaCNGC-Ps1 is shown as SEQ ID NO. 2.

[0014] The wheat disease resistance related protein TaCNGC-Ps1 is encoded by the wheat disease resistance related protein gene TaCNGC-Ps1 , and has an amino acid sequence shown as SEQ ID NO. 1.

[0015] Further, in the application of the wheat disease resistance related protein gene TaCNGC-Ps1 and a protein encoded by the wheat disease resistance related protein gene in breeding wheat resistant to scab and / or powdery mildew, the expression of the wheat disease resistance related protein gene TaCNGC-Ps1 is up-regulated, and the resistance of wheat to scab and / or powdery mildew is improved.

[0016] Compared with the prior art, the technical solution provided by the present application has at least the following beneficial effects or advantages:

[0017] The coding gene of the disease resistance related protein derived from wheat water source 11 TaCNGC-Ps1The transgenic wheat obtained by overexpressing the protein is inoculated with Fusarium graminearum PH-1, and compared with the wild-type wheat, the number of ears infected with scab is significantly reduced, and the disease index of scab is reduced. Similarly, the transgenic wheat and the wild-type wheat are inoculated with Blumeria graminis E09, and compared with the wild-type wheat, the number of spore piles on the surface of the leaves of the transgenic wheat is reduced, and the length and area of the hyphae are also reduced. The above results show that the transgenic wheat has higher resistance to Fusarium graminearum PH-1 and Blumeria graminis E09 than the wild-type wheat. The wheat disease-resistant protein TaCNGC-Ps1 and the encoding gene and application thereof provided by the present application provide a gene resource for creating wheat materials resistant to scab and powdery mildew, and will play an important role in the cultivation of plants with broad-spectrum disease resistance. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 For TaCNGC-Ps1 The results of the resistance of the overexpression wheat lines to scab. A is the disease condition of the wheat ears of each test group; B is the disease index of each test group; Fielder is the wild-type wheat plant in the control group; 17 and 29 are both test groups TaCNGC-Ps1 The overexpression wheat lines; ** is P<0.01.

[0019] Figure 2 For TaCNGC-Ps1 The results of the resistance of the overexpression wheat lines to powdery mildew. A is the disease condition of the wheat leaves of each test group; B is the length of the hyphae of the Blumeria graminis; C is the infection area of the Blumeria graminis; Fielder is the wild-type wheat plant in the control group; 17 and 29 are both test groups TaCNGC-Ps1 The overexpression wheat lines; * is P<0.05, and ** is P<0.01. DETAILED DESCRIPTION

[0020] In the following, the technical solutions of the present application are described in conjunction with the examples, but the present application is not limited to the following examples. The experimental methods and detection methods described in each example are all conventional methods unless otherwise specified; and the reagents and materials described are all commercially available unless otherwise specified.

[0021] The wheat disease-resistant protein TaCNGC-Ps1 provided by the present application is derived from wheat water source 11.

[0022] The wheat disease-resistant protein TaCNGC-Ps1 or the substance for regulating the expression of the encoding gene of the protein or the substance for regulating the activity and / or content of the protein provided in the following examples is used in the following (1)~(6):

[0023] (1) increasing the disease resistance of wheat;

[0024] (2) preparing a product for improving the resistance of wheat;

[0025] (3) breeding wheat with improved disease resistance;

[0026] (4) preparing a product for breeding wheat with improved disease resistance;

[0027] (5) improving wheat with high disease resistance or preparing a product for wheat with high disease resistance;

[0028] (3) wheat breeding.

[0029] For the wheat disease resistance related protein TaCNGC-Ps1, any one of the following is used:

[0030] (1) a protein with an amino acid sequence as shown in SEQ ID No. 1;

[0031] (2) a protein with an amino acid sequence as shown in SEQ ID No. 1, which has more than 80% identity with the protein shown in (1) and has the same function after substitution, deletion and / or addition of amino acid residues;

[0032] (3) a fusion protein with the same function obtained by connecting a tag to the N-terminus and / or C-terminus of (1) or (2).

[0033] In order to facilitate the purification or detection of the wheat disease resistance related protein TaCNGC-Ps1, a tag protein can be connected to the amino terminal end or carboxyl terminal end of the protein with an amino acid sequence as shown in SEQ ID No. 1. The tag protein includes but is not limited to the tag proteins listed in Table 1.

[0034] Table 1 Sequence of tag

[0035]

[0036] The wheat disease resistance related protein TaCNGC-Ps1 can be artificially synthesized, or a gene encoding the same can be synthesized first and then expressed biologically. The gene encoding the wheat disease resistance related protein TaCNGC-Ps1 can be obtained by deleting the DNA sequence shown in SEQ ID No. 2 and / or introducing a missense mutation, and / or connecting the coding sequence of the tag shown in Table 1 to the 5' end and / or 3' end thereof.

[0037] Those skilled in the art can easily use known methods, such as methods of directed evolution and point mutation, to mutate the nucleotide sequence encoding the wheat disease resistance related protein TaCNGC-Ps1 of the present application. Those nucleotides artificially modified to have 75% or more identity with the nucleotide sequence encoding the wheat disease resistance related protein TaCNGC-Ps1 obtained by separation of the present application, as long as they encode the wheat disease resistance related protein TaCNGC-Ps1, are also included in the present application. TaCNGC-Ps1 TaCNGC-Ps1 TaCNGC- ​​Ps1 and have the same function, are derived from the nucleotide sequence of the present application and are equivalent to the sequence of the present application.

[0038] The term "identity" used herein refers to sequence similarity to a natural nucleic acid sequence. The "identity" includes a nucleotide sequence having 75% or more, or 85% or more, or 90% or more, or 95% or more identity to the nucleotide sequence of the protein consisting of the amino acid sequence shown in SEQ ID No. 1 of the present application. The identity can be evaluated by the naked eye or computer software. Using computer software, the identity between two or more sequences can be expressed in percentage (%) which can be used to evaluate the identity between related sequences.

[0039] The above-mentioned 75% or more identity can be 80%, 85%, 90% or 95% or more identity.

[0040] The substance that regulates the expression of the gene encoding the protein TaCNGC-Ps1 can be a substance that enhances, up-regulates or increases the expression of the gene encoding the protein TaCNGC-Ps1. The substance that regulates the activity and / or content of the protein TaCNGC-Ps1 can be a substance that enhances, up-regulates or increases the activity and / or content of the protein TaCNGC-Ps1.

[0041] The disease resistance of the wheat is resistance to scab and powdery mildew. The scab can be caused by the wild type strain PH-1 of Fusarium graminearum and the powdery mildew can be caused by the strain E09 of Erysiphe graminis.

[0042] The substance in the present application is a biological material, and the biological material is any one of the following (1) to (7):

[0043] (1) a nucleic acid molecule encoding the protein;

[0044] (2) an expression cassette containing the nucleic acid molecule of (1);

[0045] (3) a recombinant vector containing the nucleic acid molecule of (1), or a recombinant vector containing the expression cassette of (2);

[0046] (4) a recombinant microorganism containing the nucleic acid molecule of (1), or a recombinant microorganism containing the expression cassette of (2), or a recombinant microorganism containing the recombinant vector of (3);

[0047] (5) A transgenic plant cell line containing the nucleic acid molecule described in (1), or a transgenic plant cell line containing the expression cassette described in (2);

[0048] (6) A transgenic plant tissue containing the nucleic acid molecule described in (1), or a transgenic plant tissue containing the expression cassette described in (2);

[0049] (7) A transgenic plant organ containing the nucleic acid molecule described in (1) or a transgenic plant organ containing the expression cassette described in (2).

[0050] Wherein, (1) the nucleic acid molecule is a cDNA molecule or DNA molecule whose coding sequence is SEQ ID No.2, encoding the amino acid sequence shown in SEQ ID No.1.

[0051] The above-mentioned biological materials include the expression cassette containing a nucleic acid molecule encoding TaCNGC-Ps1. TaCNGC-Ps1 A gene expression cassette (GCC) refers to DNA capable of expressing TaCNGC-Ps1 in host cells. This DNA may include not only promoters that initiate TaCNGC-Ps1 transcription but also terminators that terminate it. Furthermore, the expression cassette may also include enhancer sequences. Promoters that can be used in this 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 carmine synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator, and carmine and octopine synthase terminator.

[0052] Existing expression vectors can be used to construct structures containing the aforementioned... TaCNGC-Ps1Recombinant vectors for gene expression cassettes. The plant expression vectors include binary Agrobacterium vectors and vectors useful for plant microprojectile bombardment, etc. Such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1305, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA), etc. The plant expression vectors can also contain a 3' untranslated region of a foreign gene, i.e. containing a polyadenylation signal and any other DNA segment involved in mRNA processing or gene expression. The polyadenylation signal can direct polyadenylation to the 3' end of the mRNA precursor, such as the untranslated region of the 3' end of the transcription of the Agrobacterium crown gall tumor inducing (Ti) plasmid gene (such as the nopaline synthase gene Nos), the plant gene (such as the soybean storage protein gene). When using the gene construction plant expression vector of the present application, enhancers, including translation enhancers or transcription enhancers, can also be used, which can be ATG start codon or adjacent region start codon, but must be in the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The source of the translation control signal and the start codon is wide, which can be natural or synthetic. The translation initiation region can be from the transcription initiation region or the structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vector used can be processed, such as adding genes that can be expressed in plants to produce color changing enzymes or luminescent compounds (GUS gene, luciferase gene, etc.), marker genes of antibiotics (such as nptII gene conferring resistance to kanamycin and related antibiotics, bar gene conferring resistance to herbicide phosphine, hph gene conferring resistance to antibiotic hygromycin, and dhfr gene conferring resistance to methotrexate, EPSPS gene conferring resistance to glyphosate), or chemical reagent resistance marker genes (such as herbicide resistance genes), mannose-6-phosphate isomerase genes that provide the ability to metabolize mannose. For the safety of transgenic plants, no selective marker gene can be added, and the transformed plants can be directly screened under stress.

[0053] In the above biological material, the vector can be a plasmid, cosmid, bacteriophage or viral vector.

[0054] In the above biological material, the microorganism can be yeast, bacteria, algae or fungi, such as Agrobacterium.

[0055] In the above biological material, the transgenic plant cell line does not include propagation material.

[0056] The application also provides a method for cultivating transgenic wheat with improved disease resistance, comprising up-regulating or enhancing or increasing the expression amount of the gene encoding the aforementioned protein or the content of the protein in the wheat of interest to obtain the wheat with disease resistance, wherein the disease resistance of the wheat with disease resistance is higher than that of the wheat of interest.

[0057] In the method, the disease resistance of the wheat is resistance to fusarium head blight and powdery mildew. The pathogenic bacteria of the fusarium head blight is wild type strain PH-1 of Fusarium graminearum, and the pathogenic bacteria of the powdery mildew is strain E09 of Erysiphe graminis.

[0058] In the method, the up-regulation or enhancement or increase of the expression amount of the gene encoding the aforementioned protein or the content of the protein in the wheat of interest is that the gene encoding the aforementioned protein is introduced into the wheat of interest.

[0059] In the method, the transgenic plant is understood to not only include the first generation transgenic plant obtained by transforming the gene into the recipient plant, but also the offspring thereof. For the transgenic plant, the gene can be propagated in the species, or the gene can be transferred into other varieties of the same species by using conventional breeding techniques, and particularly includes commercial varieties. The transgenic plant includes seeds, callus, whole plants and cells. TaCNGC-Ps1

[0060] Example 1

[0061] This example provides the obtaining of TaCNGC-Ps1 protein and the gene encoding the same.

[0062] The 7-day-old seedlings of normal growth of wheat Shuiyuan 11 were rapidly frozen with liquid nitrogen and stored at -80°C for standby.

[0063] The total RNA of the wheat leaves was extracted by using a polysaccharide polyphenol plant RNA extraction kit (Huaiyueyang Biological Technology Co., Ltd.), and the first strand cDNA was synthesized by using reverse transcriptase XL (AMV). The cDNA was synthesized by using the SMART method, and the cDNA was used as a template, TaCNGC-Ps1 -F and TaCNGC-Ps1 -R are primers for PCR, and the PCR product is detected by 1.0% agarose gel electrophoresis to obtain a PCR product of 2070bp. The amplification primer is:

[0064] TaCNGC-Ps1 -F: 5'-ATGGACGGCCCCGGCAGCGGCCACC-3';

[0065] TaCNGC-Ps1 -R: 5'-TTAGTCTTTTGGCTTGGGCAGCAGA-3'.

[0066] ​Sequencing revealed that the PCR product contained the nucleotides shown in SEQ ID No. 2, and the gene represented by these nucleotides was named... TaCNGC-Ps1 The gene, whose nucleotide sequence is shown in SEQ ID No. 2, and whose encoded protein amino acid sequence is shown in SEQ ID No. 1, is named TaCNGC-Ps1 protein.

[0067] Example 2

[0068] This embodiment provides TaCNGC-Ps1 Application of genes in improving wheat resistance to Fusarium head blight and powdery mildew

[0069] 1. Transfer TaCNGC-Ps1 Obtaining genetically modified wheat

[0070] (1) TaCNGC-Ps1 Construction of gene overexpression vectors

[0071] The CUB vector described in this embodiment is described in the literature “Xie Shuzhang, Lei Kairong, Yang Xiaoyan, et al. Study on Agrobacterium-mediated transformation of insect-resistant gene GmCry1F into maize [J]. Southwest China Journal of Agriculture, 2015, 28(3): 962-966.

[0072] TaCNGC-Ps1 The specific construction method of the CUB carrier is as follows:

[0073] by TaCNGC-Ps1 -CUB-F and TaCNGC- -CUB-R is the primer obtained in Example 1 for amplification. Ps1 TaCNGC-Ps1 The full-length coding sequence was used as a template for PCR amplification. The amplified fragment (such as nucleotides 1-2070 in SEQ ID No. 2) was then recombined into the CUB vector digested with BamHI using the ClonExpress II One Step Cloning Kit (vazyme) to obtain the recombinant expression vector. TaCNGC- -CUB. This recombinant expression vector. Ps1 TaCNGC-Ps1 -CUB is a recombinant expression vector obtained by inserting the nucleotide sequence from SEQ ID No. 2, nucleotides 1 to 2070, into the CUB vector while keeping the remaining bases unchanged.

[0074] GGATCC -CUB-F:5'-CAGGTCGACTCTAGA GGATCC ATGGACGGCCCCGGCAGCGGCCACC-3';

[0075] TaCNGC-Psl-CUB-R: 5'-GAGCTCGGTACCCGG TaCNGC-Ps1 GTCTTTTGGCTTGGGCAGCAGAATG-3'.

[0076] Note: Underline indicates the enzyme digestion site BamH I.

[0077] (2) Transgenic wheat was obtained TaCNGC-Ps1

[0078] The recombinant expression vector was transformed into Agrobacterium tumefaciens strain EHA105 by freeze-thaw method. TaCNGC-Ps1 CUB, and the T0 generation of transgenic wheat was obtained by infecting the callus of wild-type wheat Fielder with Agrobacterium. TaCNGC-Ps1 The T0 generation of transgenic wheat was cultivated until the T3 generation of transgenic wheat was obtained. TaCNGC-Ps1 The T0 generation of transgenic wheat was cultivated until the T3 generation of transgenic wheat was obtained. TaCNGC-Ps1 The two lines of transgenic wheat overexpressing CUB were named OE.

[0079] 2. Resistance analysis of transgenic wheat to fusarium head blight and powdery mildew TaCNGC-Ps1 (1) Fusarium head blight resistance identification of transgenic wheat

[0080] TaCNGC-Ps1 A. Culture of conidia The wild-type strain PH-1 of Fusarium graminearum, the pathogen of wheat scab, was used as the strain, as described in the article "Li Yimin, et al. Functional verification of histone deacetylase genes (HDACs) of Fusarium graminearum. Northwest A&F University, 2010, Master's thesis". The strain PH-1 was cultured on PDA solid medium for activation, and a piece of solid with the Fusarium strain was taken from the above-mentioned medium using a disc sampler and placed in 200 mL of sterilized carboxymethyl cellulose (CMC) liquid medium. After 4-5 days of shaking culture at 25°C and 180 rpm, the sample was taken for microscopic examination, and the spore suspension was prepared after filtration with gauze, with a concentration of 1 x 10 5

[0081] B. Selection and preparation of plastic bags: transparent plastic bags with a size of 30 cm x 40 cm x 50 μm in length x width x thickness and plastic ties for bundling were purchased.

[0082] B. Selection and preparation of plastic bags: transparent plastic bags with a size of 30 cm x 40 cm x 50 μm in length x width x thickness and plastic ties for bundling were purchased.

[0083] ​​​C. Inoculation and bagging: At the time of anthesis, 10 μL of the spore suspension prepared in step A was dropped into the lower spikelet floret at the base of the spike. At least 30 spikes were inoculated for each line, and the inoculation site was marked with a marker pen. The transparent plastic bag prepared in step B was sprayed inside and on the inoculated spikes, and the inoculated spikes were bagged and tied with plastic twine at the lower internode. A waterproof label indicating the inoculation date was hung on the easily observable position of the inoculation row, and the bag was kept for 3 days, and then removed.

[0084] D. Observation, recording and statistics: The number of diseased spikelets was investigated and counted 7-14 days after inoculation. The resistance of the test plants to scab was evaluated by the average number of diseased spikelets, and the significance of the difference was analyzed.

[0085] The number of diseased spikelets and the results of the significance of the difference of the test plants showed that the scab phenotype of the wild type material Fielder had spread to or more than half of the spike, while the scab phenotype of the overexpression lines, including OE17 and OE29, had spread to less than half of the spike or very little (as shown in A of FIG. 1). Figure 1 The statistical analysis of the significance of the difference showed that, compared with the wild type material Fielder, TaCNGC-Ps1 the average number of diseased spikelets of the inoculated spikelets of the overexpression lines (OE17 and OE29) was significantly reduced (P<0.01) (as shown in B of FIG. 1). Figure 1 TaCNGC-Ps1 The overexpression plants showed resistance to scab. The above data showed that TaCNGC-Ps1 the overexpression plants had strong resistance to scab. TaCNGC-Ps1

[0086] (2) Resistance of transgenic wheat to powdery mildew was identified TaCNGC-Ps1 The test plants were sprayed with the conidial suspension of the pathogen Blumeria graminis f. sp. tritici (Bgt) at the time of anthesis. The conidial suspension was prepared by mixing 100 μL of the conidial suspension of Bgt with 900 μL of sterile water. The conidial suspension was sprayed on the plants until the plants were wet. The plants were then covered with a plastic bag and kept for 24 hours. The bag was then removed, and the plants were kept in a greenhouse for 7 days. The plants were then investigated and counted, and the resistance of the test plants to powdery mildew was evaluated by the average number of diseased plants.

[0087] TaCNGC-Ps1 The test plants were sprayed with the conidial suspension of the pathogen Blumeria graminis f. sp. tritici (Bgt) at the time of anthesis. The conidial suspension was prepared by mixing 100 μL of the conidial suspension of Bgt with 900 μL of sterile water. The conidial suspension was sprayed on the plants until the plants were wet. The plants were then covered with a plastic bag and kept for 24 hours. The bag was then removed, and the plants were kept in a greenhouse for 7 days. The plants were then investigated and counted, and the resistance of the test plants to powdery mildew was evaluated by the average number of diseased plants. Figure 2 ​​​-OE (experimental group) and Fielder (control group) wheat leaves were evenly sown in 7cm×7cm square pots and cultured at 60% humidity, 22℃, and 16h / 8h (light / dark) conditions. Wheat leaf segments of 5-8cm length at the one-leaf stage were cut and placed in 1% water agar medium (with benzimidazole added to prevent leaf yellowing). After equilibration overnight under the original growth conditions, inoculation was carried out. All culture dishes containing wheat leaf segments were neatly arranged, and wheat seedlings with fresh E09 powdery mildew spores were gently shaken above them to evenly distribute the spores onto the leaf segments. Powdery mildew strain E09 is described in "Li Ying, Study on the molecular mechanism of wheat resistance to powdery mildew mediated by TaGST. Northwest A&F University, 2021, Master's thesis". After inoculation, the wheat leaf segments were cultured under the original growth conditions for another 5-10 days, during which leaves were collected at different time points for subsequent experiments. After the control leaves showed full disease, they were photographed and scanned, and subsequent biomass measurements were performed.

[0088] Phenotypic results are as follows TaCNGC-Ps1 As shown in Figure A, under the condition of powdery mildew E09 infection, a large number of spore masses were observed on the leaves of all treatments of the control group wild-type wheat Fielder, while TaCNGC-Ps1 Overexpression line OE17, TaCNGC-Ps1 The number of spore masses in wheat leaves was significantly reduced in the overexpression line OE29. Statistical analysis of mycelial length and colony area measurements of powdery mildew revealed that after inoculation with powdery mildew, Figure 2 24h colony area of ​​overexpressing plants (OE17 and OE29) Figure 2 The mycelial length (as shown in C) was significantly reduced compared to the control group, and the mycelial length at 48h and 72h was significantly reduced. Figure 2 (as shown in B) Colony area ( TaCNGC-Ps1 The values ​​shown in C (as indicated in the figure) were significantly reduced compared to the control group. This proves that... ​ The overexpressing plants showed strong resistance to powdery mildew (*P<0.05, **P<0.01).

[0089] The embodiments described above are some, but not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art through related deductions and substitutions based on the inventive concept, without inventive effort, are within the scope of protection of the present invention.

Claims

1. A method of breeding a wheat variety resistant to scab, characterized in that, Overexpression of a wheat disease resistance-related protein gene TaCNGC-Ps1 , to improve the resistance of wheat to fusarium head blight caused by fusarium graminearum The nucleotide sequence of the wheat disease resistance-related protein gene TaCNGC-Ps1 is shown as SEQ ID NO.

2.

2. The method of claim 1, wherein, comprises constructing a recombinant expression vector, transforming the recombinant expression vector into wheat; the recombinant expression vector comprises the wheat disease resistance related protein gene TaCNGC-Ps1 .

3. A method of breeding a wheat variety resistant to powdery mildew, characterized by, Overexpression of a wheat disease resistance-related protein gene TaCNGC-Ps1 , to increase resistance to powdery mildew caused by Blumeria graminis. The nucleotide sequence of the wheat disease resistance-related protein gene TaCNGC-Ps1 is shown as SEQ ID NO.

2.

4. The method of claim 3, wherein, comprises constructing a recombinant expression vector, transforming the recombinant expression vector into wheat; the recombinant expression vector comprises the wheat disease resistance related protein gene TaCNGC-Ps1 .

5. A wheat disease resistance-related protein gene TaCNGC-Ps1 and the use of the encoded protein in breeding wheat resistant to scab and / or powdery mildew, characterized in that, upregulating expression of the wheat disease resistance-related protein gene TaCNGC-Ps1 increases the resistance of the wheat to fusarium head blight caused by fusarium graminearum and / or powdery mildew caused by blumeria graminis. The nucleotide sequence of the wheat disease resistance-related protein gene TaCNGC-Ps1 is shown as SEQ ID NO. 2; The wheat disease resistance related protein TaCNGC-Ps1 is encoded by the wheat disease resistance related protein gene TaCNGC-Ps1 TaCNGC-Ps1 has an amino acid sequence as shown in SEQ ID NO. 1.

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