Wheat stem rot resistance related protein TaNDR1 and application thereof

Abstract

The invention belongs to the technical field of plant biology, and particularly relates to wheat stem rot related protein TaNDR1 and application thereof. According to the invention, a gene with a non-microspecies specific disease resistance function is cloned from wheat and is named as TaNDR1. According to the invention, a barley stripe mosaic virus induced vector for silencing the TaNDR1 gene in wheat is constructed, the expression of the TaNDR1 gene is reduced, a silent TaNDR1 wheat plant is obtained, and after the silent TaNDR1 wheat is inoculated with stem rot pathogen, the stem rot resistance of the wheat is obviously reduced. Therefore, after the expression quantity of the TaNDR1 in the susceptible wheat plant is increased, the stem rot resistance of the susceptible wheat plant can be obviously improved, so that the gene can be used for cultivating the stem rot resistant wheat.

Description

Technical Field

[0001] The invention belongs to the field of plant biotechnology, and particularly relates to a wheat stem rot resistance-related protein TaNDR1 and an application thereof. Background Art

[0002] Wheat stem rot is a soil-borne fungal disease caused primarily by fungi such as Fusarium pseudograminearum and Fusarium graminearum. It primarily affects the base of the wheat stem. Infected leaf sheaths at the base of the stem gradually turn dark brown, without cloud-like lesions. As the disease progresses, the internodes at the base of the wheat stem become infected and turn light to dark brown. When field humidity is high, pink or white mold forms on the nodes and internodes, and the stems break easily. Infection during the sowing phase can cause seed rot and death, affecting wheat seedling emergence. Furthermore, wheat plants infected with the pathogen accumulate potent carcinogenic fungal toxins such as vomitoxin (DON) and nivalenol (NIV), posing a significant threat to human and animal health.

[0003] Current control measures for wheat stem rot primarily include crop rotation (non-grass crops), selection of disease-resistant varieties, and pre-sowing seed treatment with pesticides (such as difenoconazole). Field management emphasizes deep plowing, debris removal, nitrogen fertilizer control, and appropriately delayed sowing. In the early stages of the disease, pesticides such as tebuconazole should be sprayed primarily at the stem base. Specific measures should be adjusted flexibly based on local conditions. Selecting disease-resistant varieties is currently the most cost-effective control measure, but highly resistant or immune varieties have yet to be discovered. Researchers have also identified some germplasm resources with some resistance through laboratory testing, but the overall proportion is relatively low. Numerous studies have shown that wheat resistance to stem rot is controlled by multiple genes. A review of research on wheat stem rot resistance loci has mapped 140 resistance QTLs across all 21 wheat chromosomes, including 10 resistance genes. Despite the large number of resistance loci reported, the primary loci remain unclear, significantly hindering progress in improving wheat stem rot resistance. Other studies have analyzed key challenges in wheat stem rot resistance research and proposed standardized resistance identification standards and enhanced resistance research during the adult plant stage. These efforts aim to identify the primary stem rot resistance loci / genes and develop molecular markers, thereby promoting stem rot resistance improvement through marker-assisted selection. Identifying resistance genes and understanding their mechanisms are key to breeding disease-resistant varieties. Using methods such as expression profiling, association analysis between genome sequences and stem rot resistance data, generation of overexpression transgenic wheat, and analysis of loss-of-function mutants, one study cloned the wheat receptor protein-encoding gene, TaRLK-6A. TaRLK-6A interacts with the somatic embryogenesis-associated receptor protein kinase, TaSERK1, to positively regulate the expression of defense genes, including TaMPK3, TaERF3, TaDefensin, TaPR1, and TaChitinase, enhancing stem rot resistance. Researchers have also identified cell wall-related kinase genes, TaWAK-6D and TaWAK-5D600, that regulate wheat resistance to sheath blight and stem rot through similar mechanisms. The continuous discovery of wheat base rot resistance sites has laid a good foundation for improving base rot resistance. However, the current lack of key resistance genes makes it difficult to make significant progress in improving base rot resistance in the short term. Therefore, it is urgent to explore new base rot regulatory genes, analyze their disease resistance mechanisms, and develop effective functional markers to provide important genetic resources and theoretical basis for breeding base rot-resistant varieties. Summary of the Invention

[0004] The purpose of the present invention is to provide a wheat stem rot resistance-related protein TaNDR1 and its encoding gene and application.

[0005] In a first aspect, the present invention provides a wheat stem rot resistance-related protein derived from the wheat genus Triticum aestivum L., named TaNDR1, and having an amino acid sequence as shown in SEQ ID NO: 2; NDR in NDR1 is an abbreviation for non-racespecific disease resistance.

[0006] In a second aspect, the present invention provides a nucleic acid molecule encoding TaNDR1 protein, the sequence of which is shown in SEQ ID NO: 1.

[0007] In a third aspect, the present invention provides a biological material containing the nucleic acid molecule described in the second aspect, wherein the biological material is an expression cassette, a transposon, a plasmid vector, a viral vector or a host cell.

[0008] In a fourth aspect, the present invention provides use of the TaNDR1 protein described in the first aspect, the nucleic acid molecule described in the second aspect, or the biological material described in the third aspect in regulating plant resistance to stem base rot.

[0009] The present invention also provides the use of the TaNDR1 protein described in the first aspect, the nucleic acid molecule described in the second aspect, or the biological material described in the third aspect in cultivating transgenic plants with increased or decreased resistance to stem base rot.

[0010] In the above application, the plant is a monocot or a dicot. The dicot may be Arabidopsis thaliana, and the monocot may be wheat (Triticum aestivum L.), corn, etc.

[0011] In another aspect, the present invention provides a method for cultivating transgenic wheat with enhanced resistance to stem rot, comprising: overexpressing a nucleic acid molecule encoding the TaNDR1 protein in a recipient wheat plant to produce transgenic wheat; the transgenic wheat exhibits enhanced resistance to stem rot compared to the recipient wheat. The overexpression method comprises transferring a vector containing the nucleic acid molecule encoding the TaNDR1 protein into the recipient wheat plant, and screening for positive transgenic wheat. This wheat with enhanced resistance to stem rot can be used to cultivate new wheat varieties.

[0012] The present invention also provides a method for cultivating wheat with reduced resistance to stem rot, comprising: reducing the expression level and / or activity of the TaNDR1 protein in a recipient wheat to produce TaNDR1-silenced wheat; the TaNDR1-silenced wheat exhibits lower resistance to stem rot than the recipient wheat. This wheat with reduced resistance to stem rot can be used for research on wheat gene function.

[0013] In the above method, the method for reducing the expression level and / or activity of TaNDR1 protein in the recipient wheat is: transiently or stably silencing the nucleic acid molecule encoding TaNDR1 protein in the recipient wheat.

[0014] In the above method, the silencing method is: introducing the BSMV-VIGS vector system containing the silencing fragment of the nucleic acid molecule encoding the TaNDR1 protein into the recipient wheat.

[0015] The beneficial effects of the present invention are:

[0016] The present invention cloned a gene with non-race-specific disease resistance from the wheat variety Bainong's Aikang 58, named TaNDR1, and constructed a viral-inducible vector for transiently silencing the TaNDR1 gene in wheat, reducing the expression of the TaNDR1 gene and obtaining TaNDR1-silenced wheat. After inoculating the TaNDR1-silenced wheat with the stem base rot pathogen, the wheat plants showed a significant phenotype of reduced resistance to the stem base rot, indicating that inhibiting the expression of the TaNDR1 gene in wheat plants by transient silencing can reduce wheat resistance to the stem base rot. The above research results show that the TaNDR1 gene positively regulates wheat resistance to the stem base rot, and silencing the TaNDR1 gene in wheat can reduce wheat resistance to the stem base rot. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the expression analysis of TaNDR1 gene at different time points after Bainong'aikang 58 was inoculated with wheat stem rot pathogen; the horizontal axis represents the time point after Bainong'aikang 58 was infected with wheat stem rot pathogen; the vertical axis represents the relative expression level of TaNDR1 gene in the inoculated leaves;

[0018] Figure 2 The figure shows the expression detection results of the TaNDR1 gene in silenced wheat; WT represents the expression level of the TaNDR1 gene in wild-type Fielder; γ represents the expression level of the TaNDR1 gene in the control wheat plants after infection with BSMV:γ empty vector; TaNDR1 represents the expression level of the TaNDR1 gene in the wheat plants after infection with BSMV:γ-TaNDR1;

[0019] Figure 3 The phenotypes produced by VIGS-silencing wheat Fielder after inoculation with wheat stem rot pathogens are shown. WT represents the phenotype of wild-type Fielder plant leaves inoculated with wheat stem rot pathogens; γ and TaNDR1 represent the phenotypes of plant leaves infected with BSMV:γ empty control and BSMV:γ-TaNDR1, respectively, after inoculation with wheat stem rot pathogens.

[0020] Figure 4The disease index of VIGS-silenced wheat Fielder after inoculation with wheat stem rot pathogen. WT represents the disease index of wild-type Fielder plants after inoculation with wheat stem rot pathogen; γ and TaNDR1 represent the disease index of plants infected with BSMV:γ empty control and BSMV:γ-TaNDR1, respectively, after inoculation with wheat stem rot pathogen. DETAILED DESCRIPTION

[0021] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly described below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0022] Example 1: Cloning of TaNDR1 gene

[0023] 1. Obtaining cDNA

[0024] Total RNA was extracted from wheat leaves of Bainong Aikang 58 (a commercially available variety highly susceptible to stem base rot at the seedling stage) using the instructions in the Plant RNA Extraction Kit (Vazyme). The resulting RNA was reverse transcribed using polyT primers to generate cDNA.

[0025] 2. Cloning of TaNDR1 gene using information from Chinese Spring database

[0026] Using the cDNA obtained in step 1 as a template, primers P1 and P2 designed based on the TraesCS4B02G115700 (TaNDR1) transcript sequence from the Chinese Spring website (https: / / plants.ensembl.org / Triticum_aestivum / Info / Index) were used for PCR amplification using a high-fidelity enzyme to obtain a PCR amplification product; the primer sequences are as follows:

[0027] Primer P1: 5'-GCCATGGAGGCCGAATTCATGCTCCGCCCCACGCACCC-3';

[0028] Primer P2: 5′-CCGCTGCAGGTCGACGGATCCCTAGACCTCGACGTGGCAGT-3′.

[0029] The PCR amplification reaction conditions were as follows: 95°C for 5 minutes; then 95°C for 15 seconds, 57°C for 15 seconds, and 72°C for 1 minute, for 32 cycles; and finally 72°C for 5 minutes.

[0030] 3. Electrophoresis and sequencing

[0031] The PCR amplification product obtained in step 2 above was ligated to the pMD-19T vector (TaKaRa) for sequencing. The sequencing results were spliced using DNAman software to obtain the full-length ORF of the TaNDR1 gene. The ORF sequence of the TaNDR1 gene is shown in SEQ ID NO: 1, and the amino acid sequence encoded by it is shown in SEQ ID NO: 2.

[0032] Example 2: Expression Analysis of TaNDR1 in Bainong Aikang 58

[0033] 1. Experimental methods:

[0034] Take the seeds of wheat material Bainong Aikang 58, soak them in clean water at room temperature for 24 hours, pour out the liquid after the seeds are imbibed, keep them moist at room temperature for 24 hours, and plant them in pots after they turn white. Grow in a light incubator at 18℃ / 10h, 22℃ / 14h and 70% humidity. After the seeds germinate for 3 days, remove the weak seedlings with inconsistent growth status, and infect them with fresh stem base rot pathogens collected from the field. Take the wheat seedling leaves infected for 0h, 12h, 24h, 48h, and 72h respectively and quick-freeze them in liquid nitrogen. Cut 3 leaves of plants at each time point. Grind the above leaf materials separately, extract total RNA, and reverse transcribe with the kit HiScriptIII 1st Strand cDNA Synthesis Kit (+gDNA wiper) (Vazyme) to obtain cDNA. Use the cDNA as a template and use primer P3 (5'- GTTCCAGACCGCCACCTA The TaTublin fragment was amplified using primers P1 (5'-CGACGCTAACACTTATTACCAC-3') and P4 (5'-CGACGCTAACACTTATTACCAC-3') as an internal reference gene, and the TaNDR1 fragment was amplified using primers P5 (5'-GTGGAACTGGCTCTGGC-3') and P6 (5'-CGCTCAATGTCAAGGGA-3') to analyze the expression of TaNDR1.

[0035] PCR reactions were performed on a real-time fluorescence quantitative PCR instrument (Roche, Germany) and fluorescence was detected. A 20-μL PCR reaction system contained 10 μL of 2×Q3 SYBR qPCR Master Mix (Universal), 0.4 μL each of primers P3 and P4, 1 μL of cDNA template, and 8.2 μL of ddH2O. Amplification conditions were as follows: constant temperature at 95°C for 30 seconds, followed by 40 cycles of 95°C for 10 seconds and 60°C for 30 seconds, and a melting step at 95°C for 15 seconds, 60°C for 1 minute, and 95°C for 15 seconds. After the reaction, melting curves were determined. Gene expression levels were analyzed using GraphPad software.

[0036] 2. Experimental results

[0037] The results of real-time fluorescence quantitative PCR were as follows Figure 1 As shown. Figure 1 The results show that in wheat variety Bainong'aikang 58, TaNDR1 expression decreased significantly 12 hours after induction by the root rot pathogen, then increased significantly 48 hours later and returned to its original level 72 hours later. This suggests that TaNDR1 gene expression can respond to the induction of the root rot pathogen and is downregulated when infected by the pathogen.

[0038] Example 3: Acquisition of TaNDR1-silenced wheat and study of its resistance to stem rot

[0039] 1. Obtaining gene silencing fragments:

[0040] The PCR amplification product obtained in step 2 of Example 1 was ligated into the pMD-19T vector. Using the pMD-19T vector containing the ORF sequence of TaNDR1 as a template, PCR amplification was performed using primers P7 and P8 to obtain the target gene fragment, which was designated as TaNDR1 (VIGS) and used to construct a VIGS silencing vector.

[0041] Wherein, the nucleotide sequences of the above primers P7 and P8 are as follows:

[0042] P7: 5'-TAGCTGAGCGGCCGCGCCTCCTACAAGTAC-3';

[0043] P8: 5'-TAGCTGATTAATTAAGAACCTGAGCCCGTT-3'.

[0044] The PCR amplification reaction system was as follows: 1 μL plasmid template (30 ng / μL), 2 μL upstream primer P7, 2 μL downstream primer P8, 25 μL PCR mix, and water was added to 50 μL.

[0045] PCR reaction conditions were as follows: pre-denaturation at 95°C for 5 minutes, followed by 30 cycles of denaturation at 95°C for 10 seconds, annealing at 55°C for 15 seconds, and extension at 72°C for 20 seconds, followed by an additional extension at 72°C for 5 minutes, and storage at 4°C. PCR amplification products were recovered after detection of amplified bands by 1.5% agarose gel electrophoresis.

[0046] 2. Construction of BSMV recombinant viral vector:

[0047] According to conventional molecular biological methods, the silencing fragment TaNDR1 (VIGS) obtained in step 1 above was reversely inserted into the BSMV-VIGS viral vector γ by homologous recombination, while keeping other sequences of the BSMV-VIGS viral vector γ unchanged to obtain the recombinant vector γ-TaNDR1.

[0048] 3. BSMV-VIGS vector system:

[0049] BSMV-VIGS viral vectors α, β and γ vectors together constitute the viral vector system BSMV0.

[0050] BSMV-VIGS viral vectors α, β and recombinant vector γ-TaNDR1 together constitute the viral silencing vector system BSMV:γ-TaNDR1 that can silence the TaNDR1 gene.

[0051] The BSMV-VIGS viral vectors α and β and the vector γ-PDS together constitute the viral silencing vector system BSMV:γ-PDS that can silence the TaPDS gene. The γ-PDS is derived from Scofield Laboratory (Scofield, SR, Huang, L., Brandt, AS, & Gill, BS (2005). Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway. Plant physiology, 138(4), 2165-2173. https: / / doi.org / 10.1104 / pp.105.061861). It contains a 185bp conserved fragment of the barley phytoene dehydrogenase gene (PDS), which can be used as a positive control for gene silencing. After silencing of the gene, the leaves of the plant show a whitening phenomenon, which can be used to visually detect whether the silencing of the VIGS system is effective.

[0052] 4. BSMV in vitro transcription:

[0053] (1) Linearization of the vector

[0054] The α chain, γ chain, recombinant vector γ-TaNDR1 and recombinant vector γ-PDS of BSMV viral vector were digested with MluⅠ, and the β chain of BSMV viral vector was digested with SpeⅠ to obtain linearized plasmids.

[0055] (2) The linearized plasmid obtained in step (1) was used as a template for in vitro transcription to obtain viral vectors α, β, γ, γ-TaNDR1, and γ-PDS transcribed into RNA in vitro. The in vitro transcription reaction was performed according to the instructions of the mMESSAGEmMACHINE T7 invitro transcription kit (Invitrogen). The transcription reaction system and conditions were as follows: 10.0 μL of the total reaction system, including 3 μL of the linearized plasmid, 1 μL of 10× Reaction Buffer, 5 μL of 2× NTP / CAP, and 1 μL of Enzyme Mix. The reaction was carried out in a PCR instrument at 37°C for 2 h, and the transcription product was stored at -80°C for later use.

[0056] 5. Wheat Plant Cultivation and BSMV Inoculation

[0057] Select plump Fielder seeds, place them in a culture dish to absorb water for one day, pour out the water in the culture dish and rinse it clean, keep the seeds moist for one day, and after the seeds turn white, select wheat seeds with consistent growth and sow them in pots. Place the sown wheat in a light incubator for growth, with a light time of 14h light / 10h dark, a temperature of 12°C during light, and a temperature of 10°C during darkness. After three days of cultivation, remove the weak seedlings and inoculate with the pathogenic bacteria of stem base rot. When the wheat grows to two leaves and one heart, and the second leaf is consistent with the first leaf, select the wheat Fielder with consistent growth for BSMV inoculation. Water the wheat sufficiently before inoculating the virus, and apply the BSMV:γ-TaNDR1 recombinant virus vector solution to the second leaf of the wheat by friction inoculation. The inoculated wheat was placed in an incubator at 23°C and kept in the dark for 24 hours. After 24 hours, the plants were transferred to 23°C, 14h light / 10h dark conditions for growth to obtain BSMV:γ-TaNDR1 infected plants (i.e., wheat plants with silenced TaNDR1 gene).

[0058] At the same time, some plants were inoculated with BSMV:γ-PDS viral vector solution to obtain BSMV:γ-PDS transfected control plants, and some plants were inoculated with BSMV:γ viral vector solution to obtain BSMV:γ empty vector transfected control plants (γ).

[0059] The above-mentioned BSMV:γ-TaNDR1 recombinant viral vector solution is a solution obtained by mixing the in vitro transcription products according to the ratio of 10μLα, 10μLβ, 10μLγ-TaNDR1 and 225μL FES Buffer (0.1M glycine, 0.06M K2HPO4 buffer containing 1% sodium pyrophosphate, 1% macaloid, 1% celite; phosphoric acid to adjust the pH to 8.5-9.0).

[0060] The BSMV:γ-PDS viral vector solution is a solution obtained by mixing the in vitro transcription product in a ratio of 10 μL α, 10 μL β, 10 μL γ-PDS and 225 μL FES Buffer.

[0061] The BSMV:γ virus vector solution was prepared by mixing the in vitro transcription product in a ratio of 10 μL α, 10 μL β, 10 μL γ and 225 μL FES Buffer.

[0062] 6. Identification of wheat with transient TaNDR1 silencing

[0063] Approximately 14 days after viral vector application, the fourth leaf of the positive control wheat inoculated with the BSMV:γ-PDS viral vector solution exhibited a PDS albino phenotype, indicating that the gene had been silenced in wheat leaves of this age and stage. This demonstrates that the BSMV-VIGS system can be successfully applied to study gene silencing in the wheat variety Fielder.

[0064] To further verify the effect of transient silencing of the TaNDR1 gene by the BSMV-VIGS system, its expression level was detected by fluorescence quantitative PCR. The specific detection method is as follows: for plants infected with BSMV:γ-TaNDR1 and BSMV:γ empty control plants that were infected with the virus for 14 days, the fourth leaf with obvious symptoms of barley streak mosaic virus was cut to extract total RNA, and the relative expression level of the TaNDR1 gene was detected by real-time fluorescence quantitative PCR after reverse transcription. TaTublin was used as the internal reference gene. -ΔΔCt Method for calculating relative expression levels: The primers used for detecting expression levels were the same as those used in Example 2.

[0065] The relative expression level of TaNDR1 gene was detected as follows: Figure 2 As shown. Figure 2 It can be seen that the relative expression level of TaNDR1 gene in plants infected with BSMV:γ-TaNDR1 was significantly lower than that in plants infected with BSMV:γ empty vector, indicating that the silencing sequence selected in this experiment is effective.

[0066] 7. Analysis of stem rot resistance in wheat with transient TaNDR1 silencing

[0067] The BSMV:γ-TaNDR1 plants that have effectively silenced the TaNDR1 gene and the BSMV:γ empty control plants were inoculated with wheat stem rot pathogens when they were 3-5 cm tall, and the base of their stems was phenotypically identified and the disease index was calculated. The disease levels were divided into level 0 (plants are not diseased), level 1 (brown spots appear at the base of the plant stems or there are mild symptoms on the first leaf sheath), level 3 (the first leaf sheath of the plant turns brown significantly), level 5 (the second leaf sheath of the plant turns brown), level 7 (brown spots appear on the third leaf sheath of the plant, or the plant is slowed down and close to death due to the disease), and level 9 (plant death). Disease index calculation formula:

[0068] Disease index (DI) = Σ(number of plants at each disease level × representative value of disease level) / total number of plants surveyed × representative value of the most severe disease level × 100%.

[0069] Depend on Figure 3 The results showed that plants infected with the BSMV:γ empty vector showed significantly greater resistance to stem base rot than those infected with BSMV:γ-TaNDR1. Plants infected with BSMV:γ-TaNDR1 were infected from the stem base to the fourth leaf, while the control leaves infected with BSMV:γ were infected to the third leaf. This suggests that silencing the TaNDR1 gene in common wheat Fielder weakened Fielder's resistance to stem base rot.

[0070] The disease index of the phenotype was calculated. The results showed that the disease index of the wild-type control Fielder was 64%, the disease index of the BSMV:γ empty control was 58%, and the disease index of the BSMV:γ-TaNDR1 was 76%. This indicates that silencing the TaNDR1 gene in common wheat Fielder weakened Fielder's resistance to stem rot. Figure 4 shown.

[0071] The embodiments described above are only preferred embodiments of the present invention and do not limit the scope of implementation of the present invention. Therefore, any equivalent changes or modifications made according to the structure, characteristics and principles described in the patent scope of the present invention should be included in the scope of the patent application of the present invention.

Claims

1. The use of the TaNDR1 protein having an amino acid sequence as shown in SEQ ID NO: 2, a nucleic acid molecule encoding the TaNDR1 protein, and a biological material containing the nucleic acid molecule in regulating wheat stem rot resistance, wherein the biological material is an expression cassette, a transposon, a plasmid vector, a viral vector or a host cell.

2. The use according to claim 1, characterized in that The sequence of the nucleic acid molecule is shown in SEQ ID NO:

1.

3. Use of the TaNDR1 protein having an amino acid sequence as shown in SEQ ID NO: 2, a nucleic acid molecule encoding the TaNDR1 protein, and a biological material containing the nucleic acid molecule in cultivating transgenic plants with increased or decreased resistance to stem base rot.

4. The use according to claim 3, characterized in that The plant is wheat.

5. A method for cultivating transgenic wheat with improved resistance to stem rot, characterized in that: include: A nucleic acid molecule encoding TaNDR1 protein is overexpressed in recipient wheat to obtain transgenic wheat; the transgenic wheat has higher resistance to stem base rot than the recipient wheat, and the amino acid sequence of TaNDR1 protein is shown in SEQ ID NO:

2.

6. The method according to claim 5, characterized in that The overexpression method comprises the following steps: transferring a vector containing a nucleic acid molecule encoding TaNDR1 protein into a recipient wheat, and screening to obtain positive transgenic wheat.

7. A method for cultivating wheat with reduced resistance to stem rot, characterized in that: include: Reducing the expression level and / or activity of TaNDR1 protein in recipient wheat to obtain TaNDR1-silenced wheat; The TaNDR1-silenced wheat has lower resistance to stem rot than the recipient wheat, and the amino acid sequence of the TaNDR1 protein is shown in SEQ ID NO:

2.

8. The method according to claim 7, characterized in that The method for reducing the expression level and / or activity of the TaNDR1 protein in the recipient wheat is: transiently or stably silencing the nucleic acid molecule encoding the TaNDR1 protein in the recipient wheat.

9. The method according to claim 8, characterized in that The silencing method comprises the following steps: introducing a BSMV-VIGS vector system containing a nucleic acid silencing fragment encoding TaNDR1 protein into recipient wheat.

10. The method according to claim 9, characterized in that The sequence of the nucleic acid molecule is shown in SEQ ID NO: 1.

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