Wheat disease resistance related protein TaCNGC-Ps1 and gene and application thereof

Abstract

The invention discloses a wheat disease resistance related protein TaCNGC-Ps1 as well as a gene and application thereof, and belongs to the technical field of gene engineering. The amino acid sequence of the TaCNGC-Ps1 protein is as shown in SEQ ID NO. 1, and the nucleotide sequence for coding the protein is as shown in SEQ ID NO. 2. An overexpression recombinant vector containing the TaCNGC-Ps1 gene is constructed and transferred into a wheat plant to obtain transgenic wheat, and the disease resistance of the transgenic wheat to gibberellic disease and powdery mildew is remarkably enhanced. The wheat disease resistance related protein TaCNGC-Ps1 as well as the gene and the application thereof, provided by the invention, provide gene resources for wheat breeding and play an important role in cultivation work of broad-spectrum disease-resistant wheat.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology and relates to the wheat disease resistance-related protein TaCNGC-Ps1, its gene, and its applications. Background Technology

[0002] wheat scab ( Fusarium Fusarium head blight (FHB) is a fungal disease that severely damages wheat production. Chemical control is currently the main method for controlling wheat Fusarium head blight in the field. However, the long-term use of single agents has led to the pathogen developing resistance to fungicides. Breeding resistant varieties is the most economical, effective, and environmentally friendly strategy for preventing Fusarium head blight. Although breeding researchers have improved wheat resistance to Fusarium head blight through traditional breeding methods, stable resistant varieties are still lacking. Therefore, identifying Fusarium head blight resistance genes and breeding stable resistant varieties remain the main tasks of wheat Fusarium head blight resistance breeding at this stage.

[0003] Composed of obligate parasitic fungus *Brucea balsamina*, a wheat-specific fungus (from the Poaceae family). Blumeria graminis f.sp. tritici Powdery mildew, caused by *B. tumefaciens* (Bgt), is a significant disease affecting wheat production. Infection of wheat seedlings by powdery mildew fungi impairs growth and development, while infection in mature plants reduces yield. The most economical and effective measure for controlling wheat powdery mildew is the use of resistant varieties. However, due to the limited source of resistance and the rapid variation of pathogen physiological races, resistant varieties have successively lost their resistance, leading to a year-on-year increase in the affected area. Yield losses caused by powdery mildew have consistently remained between 5% and 8%. Therefore, in-depth research into genes involved in wheat resistance to powdery mildew and the discovery of new gene resources will contribute to the future development and application of molecular breeding for wheat resistance.

[0004] In nature, plants have evolved sophisticated defense mechanisms to resist infection by pathogenic microorganisms. These mechanisms include host plant-triggered immunity (PTI) and effector-triggered immunity (ETI). ETI is often accompanied by hypersensitive response (HR). HR is the most common form of disease resistance in plants, characterized by localized necrosis at the infection site, which restricts the growth of pathogens. Because MAMPs (microbial-associated molecular patterns), PAMPs (pathogen-associated molecular patterns), and DAMPs (damage-associated molecular patterns) are highly conserved, PTI is easily recognized by most pathogens, while effector proteins are highly specific, resulting in differences in ETI among different species or different physiological races. Although the defense responses induced by activated effector proteins differ from those of the host plant, both can induce a series of plant immune responses, including ion flow across the lipid membrane, increased intracellular calcium ion concentration, formation of reactive oxygen species (ROS), and activation of the MAPK (mitogen-activated protein kinase) signaling pathway. Subsequent responses include the secretion of antimicrobial proteins and cell wall lignification.

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

[0006] The purpose of this invention is to provide wheat disease resistance-related proteins to improve wheat's resistance to Fusarium head blight and powdery mildew.

[0007] In a first aspect, the present invention provides a method for breeding wheat varieties resistant to Fusarium head blight, by overexpressing wheat disease resistance-related protein genes. TaCNGC-Ps1 ; The wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.2.

[0008] Furthermore, in the method for breeding wheat varieties resistant to Fusarium head blight provided by the present invention, the method includes constructing a recombinant expression vector and transforming the recombinant expression vector into wheat; the recombinant expression vector contains the wheat disease resistance-related protein gene. TaCNGC-Ps1 .

[0009] Secondly, this invention provides a method for breeding wheat varieties resistant to powdery mildew, by overexpressing wheat disease resistance-related protein genes. TaCNGC-Ps1 ; The wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.2.

[0010] Furthermore, the method for breeding wheat varieties resistant to powdery mildew provided by the present invention includes constructing a recombinant expression vector and transforming the recombinant expression vector into wheat; the recombinant expression vector contains the wheat disease resistance-related protein gene. TaCNGC-Ps1 .

[0011] Thirdly, this invention provides a wheat disease resistance-related protein gene. TaCNGC-Ps1 The application of the encoded protein in the breeding of wheat resistant to Fusarium head blight and / or powdery mildew, the wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.2; The wheat disease resistance-related protein TaCNGC-Ps1 is derived from the wheat disease resistance-related protein gene. TaCNGC-Ps1 The encoding has an amino acid sequence as shown in SEQ ID NO.1.

[0012] Furthermore, the wheat disease resistance-related protein gene provided by this invention TaCNGC-Ps1 The application of the encoded proteins in the breeding of wheat resistant to Fusarium head blight and / or powdery mildew upregulates the wheat disease resistance-related protein genes. TaCNGC-Ps1 The expression of this substance enhances wheat's resistance to Fusarium head blight and / or powdery mildew.

[0013] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages: This invention relates to the gene encoding disease resistance-related proteins derived from wheat water source 11. TaCNGC-Ps1 Transgenic wheat was obtained by overexpression in wheat. The transgenic wheat obtained using the method described in this invention, after inoculation with *Fusarium graminearum* PH-1, showed a significant reduction in the number of ears infected with Fusarium head blight and a lower Fusarium head blight disease index compared to wild-type wheat. Similarly, after inoculating the leaves of wild-type and transgenic wheat with *Powdery mildew* E09, the number of spore masses on the leaf surface of transgenic wheat was reduced, and the length and area of ​​infected hyphae were also decreased compared to wild-type wheat. These results indicate that transgenic wheat exhibits higher resistance to both *Fusarium graminearum* PH-1 and *Powdery mildew* E09 than wild-type wheat. The wheat disease resistance-related protein TaCNGC-Ps1 and its encoding gene and applications provided by this invention offer genetic resources for creating wheat materials resistant to Fusarium head blight and powdery mildew, and will play an important role in the breeding of broad-spectrum disease-resistant plants. Attached Figure Description

[0014] Figure 1 for TaCNGC-Ps1 Results of resistance to Fusarium head blight in overexpressed wheat lines. In the figures, A represents the susceptibility of wheat ears in each experimental group; B represents the disease index of each experimental group; Fielder represents the wild-type wheat plant in the control group; 17 and 29 represent the experimental groups. TaCNGC-Ps1 Overexpression in wheat lines; ** indicates P<0.01.

[0015] Figure 2 for TaCNGC-Ps1 Results of overexpression resistance of wheat lines to powdery mildew. In the figures, A represents the susceptibility of wheat leaves in each experimental group; B represents the length of powdery mildew mycelium in each experimental group; C represents the area of ​​powdery mildew infection in each experimental group; Fielder represents the control group (wild-type wheat plants); 17 and 29 represent experimental groups. TaCNGC-Ps1 Overexpression in wheat lines; * indicates P < 0.05, ** indicates P < 0.01. Detailed Implementation

[0016] The technical solution of the present invention will be described below with reference to embodiments. However, the present invention is not limited to the following embodiments. Unless otherwise specified, the experimental methods and detection methods described in each embodiment are conventional methods; unless otherwise specified, the reagents and materials can be purchased commercially.

[0017] The wheat disease resistance-related protein TaCNGC-Ps1 provided by this invention is derived from wheat water source 11.

[0018] The wheat disease resistance-related protein TaCNGC-Ps1 or substances that regulate the expression of the gene encoding the protein or substances that regulate the activity and / or content of the protein, as provided in the following examples, are used in (1) to (6) below: (1) Increase the disease resistance of wheat; (2) Prepare products that enhance wheat resistance; (3) Cultivate wheat varieties with improved disease resistance; (4) Prepare products of wheat with improved disease resistance; (5) Improve disease-resistant wheat or prepare products of disease-resistant wheat; (3) Wheat breeding.

[0019] The wheat disease resistance-related protein TaCNGC-Ps1 is any one of the following: (1) A protein with an amino acid sequence as shown in SEQ ID No. 1; (2) A protein that has more than 80% identity with and has the same function as the protein shown in (1) obtained by substituting and / or deleting and / or adding amino acid residues of the amino acid sequence shown in SEQ ID No.1; (3) A fusion protein with the same function obtained by attaching a tag to the N-terminus and / or C-terminus of (1) or (2).

[0020] To facilitate the purification or detection of the wheat disease resistance-related protein TaCNGC-Ps1, a tag protein can be attached to the amino or carboxyl terminus of the protein, which consists of the amino acid sequence shown in SEQ ID No. 1. The tag protein includes, but is not limited to, the tag proteins listed in Table 1.

[0021] Table 1: Label Sequence

[0022] The wheat disease resistance-related protein TaCNGC-Ps1 can be synthesized artificially, or its encoding gene can be synthesized first and then expressed biologically. The encoding gene of the wheat disease resistance-related protein TaCNGC-Ps1 can be obtained by deleting and / or missense mutating the DNA sequence shown in SEQ ID No. 2, and / or attaching the coding sequence of the tag shown in Table 1 to its 5' end and / or 3' end.

[0023] Those skilled in the art can readily employ known methods, such as directed evolution and point mutation, to encode the present invention. TaCNGC-Ps1 The nucleotide sequence was mutated. Those artificially modified, having a different composition from that obtained in this invention... TaCNGC-Ps1 Nucleotides with 75% or higher nucleotide sequence identity, as long as they encode TaCNGC- Ps1 They all have the same function, are derived from the nucleotide sequence of this invention, and are equivalent to the sequence of this invention.

[0024] As used herein, the term "identity" refers to sequence similarity to a natural nucleic acid sequence. "Identity" includes nucleotide sequences that have 75% or higher, 85% or higher, 90% or higher, or 95% or higher identity with the nucleotide sequence of a protein composed of the amino acid sequence shown in SEQ ID No. 1 of this invention. Identity can be evaluated visually or using 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.

[0025] The aforementioned 75% or higher degree of identity can be 80%, 85%, 90%, or 95% or higher degree of identity.

[0026] The substance regulating the expression of the gene encoding the protein TaCNGC-Ps1 can enhance, upregulate, or increase the expression of the gene encoding the protein TaCNGC-Ps1. The substance regulating the protein activity and / or content of TaCNGC-Ps1 can enhance, upregulate, or increase the activity and / or content of the protein TaCNGC-Ps1.

[0027] The wheat exhibits resistance to Fusarium head blight and powdery mildew. Fusarium head blight can be caused by infection with the wild-type strain PH-1 of Fusarium graminearum, while powdery mildew can be caused by infection with the powdery mildew strain E09.

[0028] The substance described in this invention is a biological material, and the biological material is any one of the following (1) to (7): (1) The nucleic acid molecule encoding the protein; (2) An expression cassette containing the nucleic acid molecule described in (1); (3) A recombinant vector containing the nucleic acid molecule described in (1), or a recombinant vector containing the expression cassette described in (2); (4) Recombinant microorganisms containing the nucleic acid molecule described in (1), or recombinant microorganisms containing the expression cassette described in (2), or recombinant microorganisms containing the recombinant vector described in (3); (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); (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); (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).

[0029] 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.

[0030] The above-mentioned biological materials include the expression cassette containing a nucleic acid molecule encoding TaCNGC-Ps1. TaCNGC-Ps1A 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.

[0031] Existing expression vectors can be used to construct structures containing the aforementioned... TaCNGC-Ps1Recombinant vectors for gene expression cassettes. These plant expression vectors include binary Agrobacterium vectors and vectors suitable for plant microbombardment. Examples include pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1305, pCAMBIA1300, pBI121, pCAMBIA1391-Xa, or pCAMBIA1391-Xb (CAMBIA). The plant expression vectors may also contain the 3' untranslated region of a foreign gene, i.e., a polyadenylated signal and any other DNA fragment involved in mRNA processing or gene expression. The polyadenylated signal guides the addition of polyadenylated acid to the 3' end of the mRNA precursor; similar functions exist in the 3' untranslated regions of Agrobacterium crown gall-inducing (Ti) plasmid genes (such as the nosine synthase gene) and plant genes (such as the soybean storage protein gene). When constructing plant expression vectors using the genes of this invention, enhancers, including translational enhancers or transcriptional enhancers, can also be used. 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 correct translation of the entire sequence. The translation control signals and start codons are widely available and can be natural or synthetic. The translation initiation region can originate from the transcription initiation region or structural genes. To facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed. This can involve adding genes that can be expressed in plants, encoding enzymes or luminescent compounds that produce color changes (GUS genes, luciferase genes, etc.), antibiotic marker genes (such as the nptII gene for resistance to kanamycin and related antibiotics, the bar gene for resistance to the herbicide phosphinic acid, the hph gene for resistance to the antibiotic hygromycin, the dhfr gene for resistance to methotrexate, and the EPSPS gene for resistance to glyphosate), or chemical reagent resistance marker genes (such as herbicide resistance genes), and mannose-6-phosphate isomerase genes that provide the ability to metabolize mannose. From a safety perspective, transgenic plants can be screened directly under stress without adding any selective marker genes.

[0032] In the aforementioned biological materials, the carrier may be a plasmid, a granule, a bacteriophage, or a viral vector.

[0033] In the above-mentioned biological materials, the microorganisms may be yeast, bacteria, algae or fungi, such as Agrobacterium.

[0034] Among the aforementioned biological materials, the transgenic plant cell lines do not include propagation materials.

[0035] The present invention also provides a method for breeding transgenic wheat with enhanced disease resistance, comprising upregulating or enhancing or increasing the expression level of the encoding gene of the aforementioned protein or the content of the protein in the target wheat to obtain disease-resistant wheat, wherein the disease-resistant wheat has higher disease resistance than the target wheat.

[0036] The disease resistance of wheat described in the above method refers to resistance to Fusarium head blight and powdery mildew. The pathogen of Fusarium head blight is wild-type strain PH-1 of Fusarium graminearum, and the pathogen of powdery mildew is strain E09 of Powdery Mildew.

[0037] The method described above, which involves upregulating, enhancing, or increasing the expression level of the gene encoding the aforementioned protein in the target wheat or the content of the protein, involves introducing the gene encoding the aforementioned protein into the target wheat.

[0038] In the above method, the transgenic plant is understood to include not only the transgenic plant... TaCNGC-Ps1 The first generation of transgenic plants obtained by transforming a recipient plant with a gene, including its progeny. For transgenic plants, the gene can be propagated within the species or transferred into other varieties of the same species, particularly commercial varieties, using conventional breeding techniques. The transgenic plants include seeds, callus tissue, whole plants, and cells.

[0039] Example 1 This embodiment provides the method for obtaining the TaCNGC-Ps1 protein and its encoding gene.

[0040] Take 7-day-old wheat seedlings from the water source and freeze them quickly with liquid nitrogen. Store them at -80℃ for later use.

[0041] Total RNA was extracted from wheat leaves using a polysaccharide-polyphenol plant RNA extraction kit (Huayueyang Biotechnology Co., Ltd.). First-strand cDNA was synthesized using reverse transcriptase XL (AMV). The cDNA was synthesized using the SMART method and used as a template. TaCNGC-Ps1 -F and TaCNGC-Ps1 PCR was performed using primers -R. The PCR product was detected by 1.0% agarose gel electrophoresis, yielding a 2070 bp PCR product. The amplification primers were: TaCNGC-Ps1 -F:5'-ATGGACGGCCCCGGCAGCGGCCACC-3'; TaCNGC-Ps1 -R:5'-TTAGTCTTTTGGCTTGGGCAGCAGA-3'.

[0042] 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-Ps1The 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.

[0043] Example 2 This embodiment provides TaCNGC-Ps1 Application of genes in improving wheat resistance to Fusarium head blight and powdery mildew 1. Transfer TaCNGC-Ps1 Obtaining genetically modified wheat (1) TaCNGC-Ps1 Construction of gene overexpression vectors 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.

[0044] TaCNGC-Ps1 The specific construction method of the CUB carrier is as follows: by TaCNGC-Ps1 -CUB-F and TaCNGC-Ps1 -CUB-R is the primer obtained in Example 1 for amplification. 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-Ps1 -CUB. This recombinant expression vector. 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.

[0045] TaCNGC-Ps1 -CUB-F:5'-CAGGTCGACTCTAGA GGATCC ATGGACGGCCCCGGCAGCGGCCACC-3'; TaCNGC-Ps1-CUB-R:5'-GAGCTCGGTACCCGG GGATCC GTCTTTTGGCTTGGGCAGCAGAATG-3'.

[0046] Note: The underline indicates the restriction enzyme site BamHI.

[0047] (2) Turn TaCNGC-Ps1Obtaining genetically modified wheat Recombinant expression vector TaCNGC-Ps1 -CUB utilizes Agrobacterium to infect callus tissue of wild-type wheat Fielder to obtain T0 generation. TaCNGC-Ps1 Genetically modified wheat. Breeding T0 generation. TaCNGC-Ps1 Genetically modified wheat until the T3 generation was obtained TaCNGC-Ps1 Two wheat lines were identified, with the overexpressing wheat line denoted as OE.

[0048] 2. Transfer TaCNGC-Ps1 Resistance analysis of genetically modified wheat to Fusarium head blight and powdery mildew (1) Turn TaCNGC-Ps1 Identification of Fusarium head blight resistance in genetically modified wheat A. Conidia Culture: Wild-type strain PH-1 of *Fusarium graminearum*, the pathogen of wheat scab, was selected as the strain, as described in the thesis "Li Yimin et al. Functional Verification of Histone Deacetylase Genes (HDACs) in *Fusarium graminearum*. Northwest A&F University, 2010, Master's Thesis". Strain PH-1 was activated by culturing on PDA solid medium. A sample containing the *Fusarium graminearum* strain was taken from the medium using a disc sampler and placed in 200 mL of sterilized carboxymethyl cellulose (CMC) liquid medium. The medium was incubated at 25℃ and 180 rpm with shaking for 4-5 days. Samples were then examined under a microscope, filtered through gauze, and a spore suspension was prepared with a concentration of 1×10⁻⁶. 5 per mL.

[0049] B. Selection and preparation of plastic bags: Purchase transparent plastic bags with dimensions of 30cm×40cm×50μm (length×width×thickness) and plastic tie rope for binding.

[0050] C. Inoculation with Fusarium head blight and bagging for moisture retention: When the wheat ears are flowering, 10 μL of the spore suspension prepared in step A is dripped into the florets at the base of the lower spikelets. At least 30 ears of wheat are inoculated per line. The glumes at the inoculation site are marked with a marker. The inside of the transparent plastic bag prepared in step B and the inoculated ears are sprayed. The inoculated ears are covered with plastic bags and tied appropriately with plastic rope at the internodes below the ears. At the same time, waterproof labels indicating the inoculation date are hung in easily observable locations in the inoculated plots. The moisture-retaining bags are removed after 3 days. Wild-type material Fielder is used as the control group, and overexpressing wheat is used as the experimental group.

[0051] D. Observation, recording and statistics: The number of diseased spikelets was investigated and counted 7-14 days after inoculation. The average number of diseased spikelets was used to evaluate the resistance of the tested plants to Fusarium head blight, and the difference significance was analyzed.

[0052] The results of the number of diseased spikelets and the significance of differences in the tested plants showed that the Fusarium head blight phenotype in the wild-type material Fielder had spread to or to most of the spikelets, whileTaCNGC-Ps1 Overexpression lines, including OE17 and OE29, showed only a small or no spread of the Fusarium head blight phenotype across half an ear. Figure 1 (As shown in A). Statistical analysis of the differences showed that, compared with the wild-type material Fielder, TaCNGC-Ps1 The average number of diseased spikelets was significantly reduced in overexpression lines (OE17 and OE29) (**P<0.01). Figure 1 (As shown in B in the diagram). TaCNGC-Ps1 Plants overexpressing the gene showed resistance to Fusarium graminearum after inoculation. The above data indicate that... TaCNGC-Ps1 The overexpressed plants exhibited strong resistance to Fusarium graminearum.

[0053] (2) Turn TaCNGC-Ps1 Identification of powdery mildew resistance in genetically modified wheat The test with plump grains TaCNGC-Ps1 Overexpression transgenic materials TaCNGC-Ps1 -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.

[0054] Phenotypic results are as follows Figure 2 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, TaCNGC-Ps1 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 ( Figure 2The values ​​shown in C (as indicated in the figure) were significantly reduced compared to the control group. This proves that... TaCNGC-Ps1 The overexpressing plants showed strong resistance to powdery mildew (*P<0.05, **P<0.01).

[0055] 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 for breeding wheat varieties resistant to Fusarium head blight, characterized in that, Overexpression of wheat disease resistance-related protein genes TaCNGC-Ps1 ; The wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.

2.

2. The method according to claim 1, characterized in that, The process includes constructing a recombinant expression vector and transforming the recombinant expression vector into wheat; the recombinant expression vector contains the wheat disease resistance-related protein gene. TaCNGC-Ps1 .

3. A method for breeding wheat varieties resistant to powdery mildew, characterized in that, Overexpression of wheat disease resistance-related protein genes TaCNGC-Ps1 ; The wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.

2.

4. The method according to claim 3, characterized in that, The process includes constructing a recombinant expression vector and transforming the recombinant expression vector into wheat; the recombinant expression vector contains the wheat disease resistance-related protein gene. TaCNGC-Ps1 .

5. Wheat disease resistance-related protein genes TaCNGC-Ps1 Its encoded protein and its application in the breeding of wheat resistant to Fusarium head blight and / or powdery mildew, characterized in that, The wheat disease resistance-related protein gene TaCNGC-Ps1 The nucleotide sequence is shown in SEQ ID NO.2; The wheat disease resistance-related protein TaCNGC-Ps1 is derived from the wheat disease resistance-related protein gene. TaCNGC-Ps1 The encoding has an amino acid sequence as shown in SEQ ID NO.

1.

6. The application according to claim 5, characterized in that, Upregulation of the wheat disease resistance-related protein gene TaCNGC- Ps1 The expression of this substance enhances wheat's resistance to Fusarium head blight and / or powdery mildew.

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