TmNLR1 gene related to wheat powdery mildew resistance and application thereof

Abstract

The invention discloses a TmNLR1 gene related to wheat powdery mildew resistance and application of the TmNLR1 gene, and belongs to the technical field of molecular breeding. The CDS sequence of the TmNLR1 gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the protein coded by the gene is as shown in SEQ ID NO. 2. By overexpressing the TmNLR1 gene, the resistance of wheat to powdery mildew can be improved. According to the invention, a functional gene TmNLR1 positioned at a PmNJ3946 site in a 3AS chromosome interval is identified for the first time, and it is proved that the functional gene TmNLR1 can inhibit powdery mildew aspirator formation and improve resistance. The gene is a wheat endogenous gene and is high in safety, disease-resistant gene resources are enriched, a new target is provided for molecular breeding, and efficient cultivation of wheat disease-resistant varieties is facilitated.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of molecular breeding, and in particular to a TmNLR1 gene related to wheat powdery mildew resistance and application thereof. BACKGROUND

[0002] Wheat powdery mildew is an important disease caused by Blumeria graminis f. sp. tritici (Bgt), which seriously restricts the yield of wheat. Cultivating and utilizing disease-resistant varieties is the most economical and effective way to control the disease. However, due to the rapid population variation and continuous evolution of pathogenic types of the pathogen, many identified powdery mildew resistance genes gradually lose resistance in production application, and the problem of single resistance source is increasingly prominent, resulting in shortened resistance life of wheat varieties and increased risk of disease prevalence. Therefore, continuously exploring new genes with novel resistance mechanism and broad-spectrum and durable resistance has become an important task for wheat powdery mildew breeding.

[0003] At present, more than 140 powdery mildew resistance genes have been located and named in wheat and its close relatives, of which 24 have been successfully cloned. According to the structural characteristics of the encoded proteins, these genes are mainly divided into three categories: NLR proteins encoding CC-NBS-LRR type, kinase proteins, and transport proteins. Although a certain number of disease resistance genes have been cloned, the PmNJ3946 gene located on the 3AS chromosome of wheat has not been cloned, and its molecular mechanism is unknown, which limits the precise use of the gene in breeding. In particular, excellent resistance gene resources derived from cultivated einkorn wheat (Triticum monococcum) have not been fully explored. Only six powdery mildew resistance genes, Pm1b, PmNCA4, Mlm2033, pm2026, Pm4d, and TmPm3, have been identified from this species, of which PmNJ3946 located on 3AS has been preliminarily located, but its candidate gene has not been determined, and there is a lack of systematic functional verification, which directly affects its application potential in molecular marker-assisted selection. SUMMARY

[0004] The purpose of the present application is to provide a TmNLR1 gene related to wheat powdery mildew resistance and application thereof, in order to solve the problems existing in the prior art. The wheat powdery mildew resistance gene TmNLR1 is cloned, which is confirmed to be able to inhibit the formation of haustoria of powdery mildew and improve resistance. The gene is an endogenous gene of wheat, has high safety, enriches the resistance gene resources, provides a new target for molecular breeding, and is helpful for efficient breeding of wheat disease-resistant varieties.

[0005] To achieve the above purpose, the present application provides the following scheme:

[0006] The application provides a TmNLR1 gene related to wheat powdery mildew resistance, wherein a CDS sequence of the TmNLR1 gene is shown as SEQ ID NO. 1.

[0007] The application also provides a protein encoded by the TmNLR1 gene, wherein an amino acid sequence of the protein is shown as SEQ ID NO. 2.

[0008] The application also provides a recombinant vector containing the TmNLR1 gene.

[0009] The application also provides application of the TmNLR1 gene, the protein or the recombinant vector in improving wheat powdery mildew resistance, and overexpression of the TmNLR1 gene can improve the wheat powdery mildew resistance.

[0010] Optionally, the overexpression of the TmNLR1 gene improves the wheat powdery mildew resistance by inhibiting the formation of a haustorium of the powdery mildew.

[0011] Optionally, the powdery mildew includes Blumeria graminis f. sp. tritici.

[0012] The application also provides application of the TmNLR1 gene, the protein or the recombinant vector in cultivating wheat varieties with strong powdery mildew resistance, and overexpression of the TmNLR1 gene in the wheat can improve the powdery mildew resistance of the obtained wheat.

[0013] Optionally, the overexpression of the TmNLR1 gene improves the wheat powdery mildew resistance by inhibiting the formation of a haustorium of the powdery mildew.

[0014] Optionally, the powdery mildew includes Blumeria graminis f. sp. tritici.

[0015] The application discloses the following technical effects:

[0016] The present application provides a wheat powdery mildew resistance related gene TmNLR1 and application thereof, effectively solves the problems of insufficient wheat disease resistance gene resources and low efficiency of function verification. On the one hand, the present application first clones the candidate gene TmNLR1 of PmNJ3946 located in the 3AS chromosome interval, and determines its structural characteristics and pathogen-induced expression pattern through fine mapping and expression analysis, thereby providing a new endogenous gene resource for wheat powdery mildew resistance breeding, enriching the disease resistance gene library, and helping to broaden the genetic basis of disease resistance. On the other hand, function verification shows that overexpression of TmNLR1 can significantly inhibit the formation of haustoria of powdery mildew and improve the resistance of wheat to powdery mildew. The gene is derived from wheat itself, and has the characteristics of high application safety and easy breeding utilization. The present application not only identifies a functional gene of PmNJ3946 site, but also provides a new effective gene resource for wheat disease resistance variety breeding, and helps to promote the precision and efficiency of powdery mildew resistance breeding. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description only constitute some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of these drawings.

[0018] Figure 1 Fine mapping of PmNJ3946;

[0019] Figure 2 TmNLR1 expression level detection;

[0020] Figure 3 Growth of powdery mildew in GUS positive cells in disease resistance interaction (a) and disease susceptibility interaction (b); app represents appressorium; co represents conidium; ha represents haustorium; hy represents hypha; and pp represents penetration peg. DETAILED DESCRIPTION

[0021] The various exemplary embodiments of the present application will now be described in detail, which should not be considered as limiting the present application, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present application.

[0022] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. Additionally, for a range of values of, for example, concentrations, solvent amounts, and the like, there are intended to be included in the present application each and every intermediate value of the range. For example, a range of 1 to 2 includes each and every intermediate value between and including 1 and 2. These intermediate values can be independently combined with other dependent values.

[0023] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned herein are incorporated by reference to disclose and describe in detail the methods and / or materials associated with the documents. In the case of conflict between the present specification and any document incorporated by reference, the present specification will control.

[0024] Many modifications and variations of the present application described in the specific embodiments of the application can be made by those skilled in the art without departing from the spirit or scope of the application. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application. The specification and examples are illustrative only.

[0025] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean including, but not limited to.

[0026] The recombinant inbred line NJ3946 used for fine mapping of PmNJ3946 in the embodiments of the present application is a powdery mildew resistant line derived from the cross of einkorn wheat TA2032 and M389, and the flanking molecular markers Xbarc294 and Xwgrc5153 are polymorphic markers used for mapping the powdery mildew resistance gene PmNJ3946 in wheat. The above wheat line and molecular markers are disclosed in the literature "P Wang, et. Identification and fine mapping of PmNJ3946 for powdery mildew resistance in einkorn wheat. DOI: 10.1016 / j.cj.2023.05.010".

[0027] Example 1 Fine mapping of PmNJ3946 and screening of TmNLR1

[0028] To achieve efficient screening and identification of the PmNJ3946 candidate gene, this embodiment employs a combined approach of genetic population construction, molecular marker development, phenotypic identification, and transcriptome analysis to gradually narrow down the candidate region and ultimately identify the candidate gene TmNLR1. The specific steps are as follows:

[0029] 1. Genetic population construction and recombinant screening

[0030] By crossing the cultivated wheat recombinant inbred line NJ3946 carrying the powdery mildew resistance gene PmNJ3946 with a susceptible parent, a secondary F2 segregating population containing 1042 individual plants was constructed.

[0031] The population was initially screened using the developed lateral molecular markers Xbarc294 and Xwgrc5153, and a total of 63 individual plants that underwent recombination in the target region were identified.

[0032] 2. Fine positioning and interval narrowing

[0033] Five pairs of polymorphic molecular markers were further developed within the Xbarc294 and Xwgrc5153 regions (all molecular markers are published in the literature "P Wang, et. Identification and fine mapping of PmNJ3946 for powdery mildew resistance in einkorn wheat. DOI: 10.1016 / j.cj.2023.05.010") to genotype the above 63 recombinants, classifying them into 17 recombination types based on the distribution of recombination breakpoints.

[0034] Powdery mildew resistance phenotypes were identified in all recombinants and parents: Physiological race Bgt38 was inoculated by shaking fresh powdery mildew spores from the leaves of the susceptible control variety Sumai 3 onto the recombinants to be identified. After the susceptible controls developed full disease, a survey was conducted; plants without obvious spore masses were classified as resistant, while those with prominent spore masses were classified as susceptible.

[0035] Genetic mapping was performed using genotype and phenotypic data, ultimately locating PmNJ3946 between molecular markers Xwgrc5182 and CHS21_3A008939814, and co-segregating with markers CHS21_3A008764562 and CHS21_3A008915069. Figure 1 This location interval corresponds to the 268 kb physical region of the reference genome of cultivated wheat TA10622.

[0036] 3. Intra-regional gene analysis and preliminary screening

[0037] Gene annotation analysis was performed on the above 268 kb interval, and a total of 9 high-confidence genes were annotated.

[0038] RNA-seq transcriptome sequencing was performed on leaf tissues from disease-resistant and disease-susceptible pools (each composed of an equal mixture of 30 homozygous disease-resistant or disease-susceptible F2 single plants). The results showed that only two genes were expressed in this region, annotated as cell wall synthase interaction protein (Gene A) and NLR-type disease resistance protein (Gene B, i.e., TmNLR1).

[0039] 4. Sequence analysis and exclusion of candidate genes

[0040] Comparing RNA-seq data from the disease-resistant pool and the disease-susceptible pool revealed that both genes exhibited single nucleotide polymorphisms (SNPs) in their coding regions, resulting in alterations in their amino acid sequences.

[0041] To further differentiate candidate genes, the genome sequences of the published susceptible wheat varieties Fielder and Kenong 9204 were compared. It was found that the cell wall synthase interaction protein in the disease-resistant pool was completely identical to the sequence in these two susceptible varieties, thus ruling out its possibility as a candidate gene for PmNJ3946.

[0042] 5. Validation of candidate gene expression

[0043] The expression dynamics of TmNLR1 after inoculation with powdery mildew were detected using real-time quantitative PCR (qPCR). The wheat housekeeping gene TaActin was used as an internal control, and the SYBR Green fluorescent dye assay was employed. Results showed that TmNLR1 expression was significantly upregulated after powdery mildew inoculation. Figure 2 ), exhibiting a typical pathogen-induced expression pattern.

[0044] 6. Identification of candidate genes

[0045] Based on the combined results of region co-segregation, gene expression pattern and sequence variation analysis, TmNLR1, annotated as an NLR-type disease resistance protein, was identified as a candidate gene for PmNJ3946 for subsequent functional verification experiments.

[0046] Example 2 Cloning and Sequence Analysis of TmNLR1 Gene cDNA Sequence

[0047] The purpose of this embodiment is to clone the complete cDNA sequence of the TmNLR1 gene from the disease-resistant material NJ3946 and perform bioinformatics analysis on it, providing a sequence basis for functional verification and subsequent applications of this gene. The specific steps are as follows:

[0048] 1. RNA extraction and cDNA synthesis

[0049] Take about 100 mg of young leaf tissue from the disease-resistant material NJ3946 during the seedling stage, quick-freeze it with liquid nitrogen, and then grind it into a fine powder.

[0050] Total RNA was extracted using the Invitrogen Trizol kit, following the manufacturer's instructions. The extracted RNA was treated with DNase I (RNase-free), and the concentration and purity were determined using a NanoDrop 2000, ensuring an A260 / A280 ratio between 1.8 and 2.0 and an A260 / A230 ratio greater than 2.0.

[0051] Take 1 μg of total RNA and synthesize first-strand cDNA using the reverse transcription kit from TransGen Biotech (Beijing). Store the resulting cDNA at -20℃ for later use.

[0052] 2. Specific primer design and PCR amplification

[0053] Based on the TmNLR1 genome sequence obtained in Example 1, specific amplification primer pair P1 was designed upstream of the predicted open reading frame (ORF) start codon (ATG) and downstream of the stop codon:

[0054] Forward primer F: 5′-ATGGCAGAGGGTGTTGTAGCGTTG-3′ (SEQ ID NO.3)

[0055] Reverse primer R: 5′-CTAATTAAAGTCAACCCATCTCA-3′ (SEQ ID NO.4)

[0056] PCR reaction system (25 μL): 10 ng cDNA template, 5 pmol each of forward primer F and reverse primer R, 5 nmol each of dNTP mixture (2.5 mM), 0.5 U Taq DNA polymerase (5 U / μL), 2.5 μL 1× PCR buffer, and ddH2O added to a total volume of 25 μL.

[0057] PCR amplification program settings: denaturation at 94℃ for 3 minutes; 36 cycles, denaturation at 94℃ for 30 seconds, annealing at 60℃ for 30 seconds, extension at 72℃ for 3 minutes; final extension at 72℃ for 5 minutes.

[0058] 3. PCR product purification and cloning

[0059] The PCR product was detected by 1% agarose gel electrophoresis. The expected amplified band size was approximately 2733 bp. The product was then purified by gel extraction.

[0060] The purified PCR product was ligated with the pMD19-T vector (TaKaRa). The ligation system was 10 μL: 1 μL pMD19-T vector, 4 μL purified PCR product, 1 μL ligase, 2 μL buffer, and ddH2O added to 10 μL. The ligation reaction was carried out in a 37℃ water bath for 30 min.

[0061] 4. Transformation and Sequencing

[0062] All of the above ligation products were transformed into Escherichia coli DH5α competent cells and subjected to heat shock: heat shock at 42℃ for 90 s, followed by ice bath for 2 min.

[0063] After transformation, the bacterial culture was obtained by culturing in LB liquid medium, spread on LB solid plates containing ampicillin, and incubated upside down.

[0064] White single colonies were selected for colony PCR verification, and positive clones were sent to Beijing Qingke Biotechnology Co., Ltd. for Sanger paired-end sequencing.

[0065] 5. Sequence Analysis and Naming

[0066] After splicing and correction, the sequence obtained from sequencing yielded a full-length cDNA sequence of 2733 bp (SEQ ID NO.1), containing a complete open reading frame.

[0067] Analysis using the NCBI ORF Finder tool confirmed that the ORF encodes a protein consisting of 910 amino acids, with a predicted molecular weight of approximately 103.5 kDa and a theoretical isoelectric point (pI) of 6.76. The amino acid sequence is shown in SEQ ID NO.2.

[0068] The gene was named TmNLR1.

[0069] SEQ ID NO.1:

[0070]

[0071] SEQ ID NO.2:

[0072] MAEGVVALLIAKLGFALAKEAATFGASLLCKEASALKGLFGEIREAKEELESMQAYLQGAERFKDTDETTGIFVNKVRGFAFEIEDVVDEFTYKLEDTHGGFTTKMKKRVKHIKAWRRLTLKLQDIKGRLQGADRRKVRYDMRGIDRGGCNNVQSRSADHSLNPPREEDLVGIEENKDKLMHWLVGDLEEQGSKIATIWGMGGVGKTTLVHHVYKAVKMDFAKYAWITVSSSYQVEDLLRQMATELGVAIGLADANRNLVEVIHNHLQGSEYLIVLDDVWHLDVWFKVRNAFPTESTSRFVITTRIQEVALLATKNCTIKLEPLERHHAWQLFCNEAFWNNEKKTCPDELEILAQMFLDKCGGLPIAIACVGRLLSCRHPTYSEWKSLYKELELQLSNNVILDVNAVLKVSLEDLPTDLKNCFLYCTIFPEDYLFQRRQVMRHWIASGFIKEAGNKTLDEVAEGYLNKLVNRSLLQVVERNRAGQVYRCRMHDILHVLALAKSEEESFCHVYNGLTPFSTEKTRRLSIQSTNVEQLAPLLCATSLRSLHVFESHLRIDSLEAFLKPFNLLSTLDLQGVQIKKLPKIVFNLFNLRFLGLRDTHIEYLPKEIGRLQNLEVLDAYNSMLSALPVQVATLRKLKYLYVVTIPAGADERVLTFEGIQVPKGIGNLTDLLALQLIEASSEVLFQLGYLTKMRSFSITKVRSGHWADLCGAIAKMVHLVHVTIHSLDQREVLQLESLCLPPTVSEVVIYAQLDQRVLPQFVSSSSKLINLTGLHLGWSKLDENSFACLLGLHGLVTLVLNKAYDGKELNFPATSLPKLKYLRIWDAPNLSRVTIEQRAMQNIVQLILTDCPELKHLPHGIEHLRTLEYLKLCDISEELTRKLQLNEESKECNEDWMKISHVRWVDFN。

[0073] Example 3: Validation of TmNLR1's resistance to powdery mildew based on a transient overexpression system

[0074] This embodiment aims to verify whether the TmNLR1 gene can enhance wheat resistance to powdery mildew by using a transient overexpression system in wheat leaves. The specific steps include vector construction, gene gun-mediated transient transformation, powdery mildew inoculation, histochemical staining, and microscopic observation. The pAT2 vector used for the transient overexpression was modified from the pMD19-T vector and contains a Ubi+Nos expression cassette. The pAT2 vector is disclosed in invention patent application number 202411937862.7.

[0075] 1. Construction of the overexpression vector pAT2-TmNLR1

[0076] Using the clone containing the complete coding sequence (CDS) of TmNLR1 obtained in Example 2 as a template, primer P2 with homologous recombination arms was designed:

[0077] Forward primer F: 5'-GTGTTACTTCTGCAGATGGCAGAGGGTGTTGTAGCGTTG-3' (SEQ ID NO.5);

[0078] Reverse primer R: 5'-TGAACGATCCTGCAGCTAATTAAAGTCAACCCATCTCA-3' (SEQ ID NO.6);

[0079] The PCR amplification system and conditions were the same as in Example 2, and TmNLR1 fragments with homologous sequences of the pAT2 vector at both ends were obtained.

[0080] The PCR product containing the homologous arm was ligated with linearized pAT2 via homologous recombination to obtain the recombinant plasmid pAT2-TmNLR1.

[0081] 2. Gene gun micro-projectile preparation and bombardment

[0082] Take 2 mg of tungsten powder with a diameter of 1.1 μm, wash it three times each with 70% ethanol and sterile ddH2O, and resuspend it in 50% glycerol.

[0083] Take 5 μg of recombinant plasmid pAT2-TmNLR1 and mix it with an equimolar amount of pUC19-GUS (CaMV 35S::GUS) reporter plasmid. Add 18 μL of the above suspended microparticles, mix well, and then add 250 μL of 2.5 M CaCl2 and 50 μL of 0.1 M spermidine sequentially. Vortex for 15 min, let stand at room temperature for 2 min, centrifuge at 10000 rpm for 5 s, discard the supernatant, wash once with 70% ethanol, wash twice with anhydrous ethanol, and finally resuspend in 45 μL of anhydrous ethanol. The plasmid used for the disease control is a mixture of pAT2 and an equimolar amount of pUC19-GUS (CaMV 35S::GUS) reporter plasmid. The microparticle preparation method is the same as above.

[0084] Select healthy, disease-susceptible wheat seedlings of the one-leaf stage, cut off a leaf segment about 5 cm from the tip of the first leaf, place it with the underside facing up on MS medium containing 0.6% agar.

[0085] The bombardment was performed using a PDS-1000 / He gene gun with the following parameters: ruptured membrane pressure: 1100 psi; bombardment distance: 9 cm; vacuum: 28 inch Hg; one bombardment per dish.

[0086] 3. Inoculation and culture of powdery mildew

[0087] Ten hours after bombardment, freshly propagated Bgt physiological race Bgt38 spores were taken and evenly shaken onto the surface of the bombarded leaves.

[0088] The inoculated leaves were placed in an incubator at 20±1℃, 70% relative humidity, and a photoperiod of 16 h light / 8 h darkness for 60 h.

[0089] 4. Histochemical staining and microscopic observation

[0090] GUS staining: Take the inoculated leaves and immerse them in GUS staining solution (0.2 M Na2HPO4 / NaH2PO4 buffer pH 7.0, 10 mM EDTA, 5 mM potassium ferricyanide, 5 mM potassium ferrousyanide, 0.1 mg / mL X-Gluc, 0.1% Triton X-100) and incubate at 37°C in the dark for 8 h.

[0091] Decolorization: Transfer the leaves to a 3:1 (v / v) ethanol:acetic acid decolorization solution and decolorize at 37°C for 30 min until chlorophyll is completely removed.

[0092] Fungal structure staining: After decolorization, the leaves were immersed in fungal staining solution (0.3% Coomassie Brilliant Blue R-250, 7.5% trichloroacetic acid, 50% methanol) for 5 min, and then gently rinsed twice with distilled water.

[0093] Microscopic observation: Leaves were placed on glass slides, and 50% glycerol was added as a flotation and preservative. GUS staining and the growth of powdery mildew hyphae and haustoria were observed using an optical microscope (Nikon ECLIPSE 80i). Cells expressing GUS and exhibiting powdery mildew haustoria growth were designated as susceptible interacting cells. Figure 3 (a) Cells expressing GUS and forming invasive nails after powdery mildew spore germination but not forming haustoria were used as disease-resistant interaction cells. Figure 3 (b) The ratio of susceptible interacting cells to the sum of susceptible and resistant interacting cells is the haustorium index (HI), which is used as an indicator of the impact of the target gene on disease resistance. Epidermal cells of Yangmai 158 leaves transformed with CaMV 35S::GUS and pAT2 were used as haustorium index control.

[0094] 5. Results

[0095] Compared with the empty vector control, the haustorium index in the leaves of Yangmai 158 overexpressing TmNLR1 was significantly lower than that in the susceptible control (see Table 1), indicating that the expression of TmNLR1 can effectively inhibit the formation of powdery mildew haustorium and enhance the resistance of wheat to powdery mildew.

[0096] Table 1. Powdery mildew haustorium index (HI) of leaves of transgenic and recipient materials (%)

[0097]

[0098] Note: ** represents P<0.01.

[0099] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A TmNLR1 gene associated with resistance to wheat powdery mildew, characterized in that, The CDS sequence of the TmNLR1 gene is shown as SEQ ID NO.

1.

2. A protein encoded by the TmNLR1 gene according to claim 1, characterized in that, The amino acid sequence of the protein is shown as SEQ ID NO.

2.

3. A recombinant vector comprising the TmNLR1 gene of claim 1.

4. The use of the TmNLR1 gene of claim 1, the protein of claim 2 or the recombinant vector of claim 3 in improving the resistance of wheat to powdery mildew. Overexpression of the TmNLR1 gene can improve the resistance of the wheat to powdery mildew.

5. Use according to claim 4, characterized in that, Overexpression of the TmNLR1 gene improves the resistance of the wheat to powdery mildew by inhibiting the formation of haustoria of the powdery mildew.

6. Use according to claim 5, characterized in that, The powdery mildew includes Blumeria graminis f. sp. tritici.

7. The use of the TmNLR1 gene of claim 1, the protein of claim 2 or the recombinant vector of claim 3 in breeding wheat varieties with high resistance to powdery mildew, characterized in that, Overexpression of the TmNLR1 gene in wheat can improve the resistance of the wheat to powdery mildew.

8. Use according to claim 7, characterized in that, Overexpression of the TmNLR1 gene improves the resistance of the wheat to powdery mildew by inhibiting the formation of haustoria of the powdery mildew.

9. Use according to claim 8, characterized in that, The powdery mildew includes Blumeria graminis f. sp. tritici.

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