Application of wheat lectin receptor kinase TaSIT2 and its encoding gene TaSIT2 in improving crop resistance to stripe rust

Abstract

This invention belongs to the fields of plant genetic engineering and crop breeding technology, specifically relating to wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 Applications in improving crop resistance to stripe rust. This invention utilizes reverse genetics to analyze and screen wheat lectin receptor kinase genes. TaSIT2 Through gene editing technology and Agrobacterium-mediated genetic transformation, we can obtain... TaSIT2 Gene-edited wheat plants showed significantly enhanced resistance to wheat stripe rust races, indicating that knockout... TaSIT2 Genes enhanced wheat resistance to stripe rust and identified wheat lectin receptor kinases. TaSIT2 Genes play a negative regulatory role in wheat resistance to stripe rust, providing new technical ideas and gene resources for breeding wheat varieties resistant to stripe rust from a molecular biology perspective, and offering more pathways for breeding varieties with long-lasting and broad-spectrum disease resistance.

Description

Technical Field

[0001] This invention belongs to the fields of plant genetic engineering and crop breeding technology, specifically relating to wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 Applications in improving crop resistance to stripe rust. Background Technology

[0002] wheat( Triticum aestivum L. Wheat is an important food crop. Wheat stripe rust is one of the most important wheat diseases. Wheat stripe rust is caused by the wheat-specific strain of *Striga styracifolium*. Puccinia striiformis f.sp. tritici , Pst Wheat stripe rust, a fungal disease caused by *Strombus heliotropium indicum*, is characterized by rapid spread, wide range of damage, and severe losses. *Strombus heliotropium indicum* is a live obligate parasite that thrives in cool, humid environments and can spread long distances via high-altitude air currents. Due to its complex life cycle, frequent variations in virulence, and the continuous emergence of new pathogenic races, controlling wheat stripe rust is extremely difficult. Currently, breeding and promoting new wheat varieties resistant to stripe rust is the most economical and environmentally friendly approach to controlling the disease. Therefore, identifying resistance-related genes, deeply analyzing the pathogenic mechanism of *Strombus heliotropium indicum* and the wheat's own resistance mechanism to stripe rust, and ultimately creating durable resistant materials are of great significance for the genetic improvement of wheat resistance to stripe rust.

[0003] Plant lectin receptor-like kinases (LecRLKs) belong to the plant membrane protein receptor kinase class. LecRLKs are mainly composed of extracellular lectin domains, transmembrane domains, and intracellular kinase domains. Based on the specificity of the extracellular domains, they are classified into G-type, L-type, and C-type. RLKs sense external signals through receptors on the cell surface, transmit them to downstream effector molecules, and initiate signal transduction. They also activate or inhibit other target proteins through phosphorylation modification, participating in the regulation of life processes such as cell growth, differentiation, immune response, neural signal transduction, and apoptosis. Therefore, RLKs play a key role in plants' response to biotic and abiotic stresses. Existing studies have confirmed that RLKs are PRRs in plant-pathogen interactions, exhibiting high sensitivity to pathogen recognition. Their extracellular lectin domains are highly variable, capable of recognizing biological stress signaling molecules from bacteria, fungi, and other organisms. They can transmit external PAMP molecular signals into the cell, thereby triggering a series of immune responses, including ROS burst, MAPK activation, callose deposition, calcium ion influx, and salicylic acid accumulation. Therefore, the role of RLKs in participating in plant responses to pathogens can be utilized to improve plant disease resistance, which is of great significance for the breeding of high-yielding and disease-resistant varieties.

[0004] TaSIT2The gene is a single copy in the wheat genome, located only on chromosome 3B. It is 2016 bp in length, encoding 671 amino acids, and has a protein molecular weight of 73.73 kDa. It belongs to the LecRLK subfamily and encodes the wheat L-type lectin domain receptor kinase SIT2 protein TaLecRLK-SIT2. However, there are few reports on TaSIT2, and its role and regulatory mechanism in wheat disease resistance are still unknown. Summary of the Invention

[0005] The purpose of this invention is to provide wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 Its application in improving crop resistance to stripe rust has enriched the new uses of wheat lectin receptor kinase TaSIT2 and its encoding gene, providing new genetic resources for breeding stripe rust-resistant crops.

[0006] To ensure a complete and unambiguous understanding of the technical solution of this invention, it should be noted that the TaSIT2 protein described in this invention is represented by "TaSIT2" in non-italicized font. TaSIT2 Genes in italic font TaSIT2 This indicates that, of course, those skilled in the art can clearly and completely understand the meaning and description of the relevant genes and their encoded proteins based on the description of this invention.

[0007] This invention provides wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 Application in improving crop resistance to stripe rust and / or breeding stripe rust-resistant crops; the amino acid sequence of the wheat lectin receptor kinase TaSIT2 is shown in SEQ ID NO:2.

[0008] Preferably, the application includes: knockout and / or knockdown in recipient plants. TaSIT2 Genes to improve crop resistance to stripe rust and / or to breed stripe rust-resistant crop varieties.

[0009] Preferably, the knockout and / or knockdown methods include CRISPR / Cas9 gene editing technology.

[0010] The present invention also provides a knockout and / or knockdown method. TaSIT2 The nucleic acid molecule of the gene includes a first specific target and a second specific target, the nucleotide sequences of which are shown in SEQ ID NO:5 and SEQ ID NO:6, respectively.

[0011] The present invention also provides a gene editing vector, comprising an initial vector and a nucleic acid molecule inserted into the initial vector, wherein the nucleic acid molecule is the nucleic acid molecule described in the above technical solution.

[0012] The present invention also provides a transformant, which includes at least one of engineered bacteria, transgenic crop cells and transgenic crop tissues, and the transformant includes the nucleic acid molecule or gene editing vector described in the above technical solutions.

[0013] This invention also provides the application of the nucleic acid molecules, gene editing vectors, or transformants described in the above technical solutions in improving crop resistance to stripe rust and / or cultivating stripe rust-resistant crop varieties.

[0014] Preferably, the crop includes wheat.

[0015] This invention also provides a method for breeding wheat resistant to stripe rust, comprising the following steps: reducing the expression level of wheat lectin receptor kinase TaSIT2 in recipient wheat or inhibiting the expression level of TaSIT2 in recipient wheat. TaSIT2 Gene expression resulted in wheat resistant to stripe rust;

[0016] The TaSIT2 The gene encodes wheat lectin receptor kinase TaSIT2, and the amino acid sequence of wheat lectin receptor kinase TaSIT2 is shown in SEQ ID NO:2.

[0017] Preferably, the reduction of the expression level of wheat lectin receptor kinase TaSIT2 in recipient wheat or the inhibition of the expression level of recipient wheat... TaSIT2 Methods for gene expression include: gene expression via Agrobacterium-mediated genetic transformation containing knockout and / or knockdown gene molecules. TaSIT2 The gene-editing vector containing the nucleic acid molecule of the gene was introduced into recipient wheat; TaSIT2 The nucleotide sequence of the open reading frame of the gene is shown in SEQ ID NO:1.

[0018] Beneficial effects:

[0019] This invention provides wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 In improving crop resistance to stripe rust, this invention reveals for the first time the role of wheat lectin receptor kinase TaSIT2 in stripe rust resistance and creates highly resistant germplasm resources through transgenic technology, providing a solution to the problem of scarce wheat stripe rust resistance resources and the difficulty in controlling wheat stripe rust. Specifically, this invention elucidates the function of wheat lectin receptor kinase TaSIT2 in the interaction between wheat and stripe rust fungus, and uses CRISPR / Cas9 gene editing technology to knock out wheat lectin receptor kinase in wheat plants. TaSIT2 Genes that enhance wheat resistance to stripe rust pathogens. The wheat lectin receptor kinase obtained using the method of this invention has been verified. TaSIT2Gene-edited wheat varieties exhibit resistance to the main prevalent races of stripe rust. This invention provides a new technical approach for breeding wheat varieties resistant to stripe rust from a molecular biology perspective, and further provides genetic resources for wheat resistance to stripe rust, offering more pathways for breeding durable and broad-spectrum resistant varieties. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0021] Figure 1 This is a flowchart illustrating the verification method for the application of wheat lectin receptor kinase TaSIT2 in the breeding and improvement of wheat rust-resistant varieties.

[0022] Figure 2 In Example 3 TaSIT2 A schematic diagram of gene editing in a gene-edited wheat plant;

[0023] Figure 3 In Example 3 TaSIT2 Phenotypic results of gene-edited wheat plants inoculated with the stripe rust-compatible race CYR32, where "Fielder" represents wild-type Fielder wheat plants; TaSIT2 KO#L2-1-1、 TaSIT2 KO#L2-2-1、 TaSIT2 KO#L4-2-6、 TaSIT2 KO#L4-3-1、 TaSIT2 KO#L5-1-7、 TaSIT2 KO#L6-2-3、 TaSIT2 KO#L6-1-6 represent respectively TaSIT2 Four strains of genetically modified wheat plants: #L2, #L4, #L5, and #L6. Detailed Implementation

[0024] This invention provides wheat lectin receptor kinase TaSIT2 and its encoding gene. TaSIT2 Application in improving crop resistance to stripe rust and / or breeding stripe rust-resistant crops; the amino acid sequence of the wheat lectin receptor kinase TaSIT2 is shown in SEQ ID NO:2.

[0025] As one implementation method, the present invention described TaSIT2 The nucleotide sequence of the gene's open reading frame (ORF) is shown in SEQ ID NO:1.

[0026] The sequences of SEQ ID NO:1 to SEQ ID NO:2 of this invention are as follows:

[0027] SEQ ID NO: 1 is as follows:

[0028]

[0029] SEQ ID NO: 2 is as follows:

[0030] MKLTMLFLVLFSFHLAAGDDVQVQFAFPGLAGANLTLDGNATVTPDGLLVLTSRKTNLKGHAFYPTPLRFRGSPGGQVHSFASSFVFAIVSDYTDFSAHGMALVVCPTADSLSSALPAGYLALLNVQNNGNASNHLFAVELDTTQNTDFQDINANHVGIDVNDLHSLQSYPTGYYLDDGGDFRNLTLFSREVMQVWVNYDGGTGQIDVTLAPIGVARPARPLVSATYNLSSVITEQAYIGFSSATGGINTRHYVLAWSFAMNGPAPAIDVAKLPKLPRFGPKPRSKVLEIVLPIATAVIILAVCTVVALLLLRHLRYAQLLEDWEVEFGPHRFSYKELYHATKGFKSKHLLGAGGFGKVYKGVLHKSKMEVAIKKISHESRQGMKEFIAEVVSIGRLRNRNLVQLLGYCRRKGELLLVYEYMPNGSLDKYLYGECDENNVHVLDWTKRLHIIKGVASGLLYIHEKWEKIVIHRDIKASNVLLDSEMNGRLGDFGLARLYDHGTDPQTTHVVGTMGYLAPELVRTGKASTLTDVFAFGAFLLEITCGRRPVSNGPQDSHEMLVDWVLDHFRRGCLAETVDVRMGGDYSVDEACRVLKLGLLCSHPFASTRPSMRHVMQYLDSDTPLPDLSSADMMSFSTMALLQNQGFDSYAISYPSSMGSIGTVSYISGGR。

[0031] As one implementation manner, the application includes: knocking out and / or knocking down in a recipient plant TaSIT2The gene is used to enhance crop resistance to stripe rust and / or to cultivate stripe rust-resistant crop varieties. As another implementation, the knockout and / or knockdown method includes CRISPR / Cas9 gene editing technology. More specifically, a gene editing vector is constructed using CRISPR / Cas9 gene editing technology, and the gene editing vector is introduced into a recipient crop via Agrobacterium-mediated genetic transformation to obtain wheat lectin receptor kinase TaSIT2 gene-edited crop varieties. As one implementation, the crop can be a gramineous crop or wheat; when the crop is wheat, the gene editing vector is introduced into the wheat embryo to cultivate... TaSIT2 Gene editing of wheat varieties can improve wheat's resistance to stripe rust.

[0032] The present invention also provides a knockout and / or knockdown method. TaSIT2 The nucleic acid molecule of the gene includes a first specific target and a second specific target, and the nucleotide sequences of the amplified first specific target and second specific target are shown in SEQ ID NO:5 and SEQ ID NO:6, respectively. As one embodiment, the first specific target and the second specific target are amplified using target primer-target-1 and target primer-target-2, respectively. The forward and reverse primers of target primer-target-1 include the nucleotide sequences shown in SEQ ID NO:7 and SEQ ID NO:8, respectively, and the forward and reverse primers of target primer-target-2 include the nucleotide sequences shown in SEQ ID NO:9 and SEQ ID NO:10, respectively.

[0033] This invention also provides a gene-editing vector, comprising an initial vector and a nucleic acid molecule inserted into the initial vector, wherein the nucleic acid molecule is the nucleic acid molecule described in the above-described technical solution. As one embodiment, the initial vector is a Cas9 plasmid vector. This invention does not specifically limit the construction method of the gene-editing vector; conventional construction methods in the art can be used.

[0034] This invention also provides a transformant, which includes at least one of engineered bacteria, transgenic crop cells, and transgenic crop tissues. The transformant comprises the nucleic acid molecule or gene editing vector described in the above-described technical solutions. The transgenic crop cells and transgenic crop tissues described in this invention are non-stable genetic cells and tissues. As one embodiment, the crop can be a gramineous crop or wheat.

[0035] This invention also provides the application of the nucleic acid molecules, gene editing vectors, or transformants described in the above-mentioned technical solutions in improving crop resistance to stripe rust and / or cultivating stripe rust-resistant crop varieties. As one embodiment, the crop can be a gramineous crop or wheat.

[0036] Specifically, the present invention also provides a method for breeding wheat resistant to stripe rust, comprising the following steps: reducing the expression level of wheat lectin receptor kinase TaSIT2 in recipient wheat or inhibiting the expression level of TaSIT2 in recipient wheat. TaSIT2 Gene expression resulted in wheat resistant to stripe rust;

[0037] The TaSIT2 The gene encodes wheat lectin receptor kinase TaSIT2, and the amino acid sequence of wheat lectin receptor kinase TaSIT2 is shown in SEQ ID NO:2.

[0038] As one implementation method, the method involves reducing the expression level of wheat lectin receptor kinase TaSIT2 in recipient wheat or inhibiting the expression level of recipient wheat lectins. TaSIT2 Methods for gene expression include: gene expression via Agrobacterium-mediated genetic transformation containing knockout and / or knockdown gene molecules. TaSIT2 The gene-editing vector containing the nucleic acid molecule of the gene was introduced into recipient wheat; TaSIT2 The nucleotide sequence of the gene's open reading frame is shown in SEQ ID NO:1. The present invention has addressed knockout and / or knockdown in the above technical solution. TaSIT2 The nucleic acid molecules of genes and gene editing vectors have been defined in detail, and will not be elaborated further.

[0039] Compared with traditional disease resistance breeding techniques, crop disease resistance genetic engineering technology can achieve targeted improvement of target traits in a shorter period of time, providing crops with more comprehensive, continuous, and broad-spectrum protection. This invention, through gene function research and genetic engineering technology, not only discovered wheat lectin receptor kinases... TaSIT2 The gene plays a negative regulatory role in the defense response against stripe rust fungus invasion of wheat, specifically by knocking out wheat lectin receptor kinase. TaSIT2 Genes can enhance wheat's resistance to stripe rust, and a wheat lectin receptor kinase resistant to wheat stripe rust has been successfully created. Figure 1 Gene-edited wheat varieties have provided valuable transgenic material for the breeding of wheat varieties resistant to stripe rust.

[0040] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0041] Unless otherwise specified, the experimental and detection methods in the following embodiments are conventional methods; the reagents and materials mentioned are commercially available unless otherwise specified; and the index data are measured using conventional methods unless otherwise specified.

[0042] Figure 1 A flowchart is provided to demonstrate the verification method for the application of the wheat lectin receptor kinase TaSIT2 in the breeding and improvement of wheat rust-resistant varieties. The verification methods used for other host plants of stripe rust are similar. TaSIT2 The methods may be identical or slightly different in technique. Embodiments of this invention provide a method for verifying the function of the wheat lectin receptor kinase TaSIT2, comprising:

[0043] S101, Acquiring wheat lectin receptor kinase TaSIT2 Gene-edited wheat: Hi-TOM gene editing sequencing was performed on the obtained TaSIT2 gene-edited wheat.

[0044] S102, T2 generation TaSIT2 gene-edited plants were inoculated with major prevalent races of stripe rust to identify the resistance of the gene-edited plants to these prevalent races and determine... TaSIT2 The stripe rust resistance of gene-edited wheat varieties.

[0045] Example 1

[0046] This embodiment provides the coding gene sequence and amino acid sequence of wheat lectin receptor kinase TaSIT2, and the steps are as follows:

[0047] Wheat lectin receptor kinases were obtained from the Ensembl Plants website (https: / / plants.ensembl.org / Triticum_aestivum / Info / Index). TaSIT2 The open reading frame (ORF) sequence of a gene and its protein sequence. Designed for amplification. TaSIT2 Primers for gene cDNA sequence amplification were used to amplify cDNA from wheat Fielder plants. TaSIT2 Genes were sequenced.

[0048] TaSIT2 The open reading frame (ORF) sequence of the gene is shown in SEQ ID NO:1, and its amino acid composition is shown in SEQ ID NO:2, used for amplification. TaSIT2 Forward primers for gene cDNA sequence TaSIT2 -cDNA-F as shown in SEQ ID NO:3, reverse primer TaSIT2 -cDNA-R is shown in SEQ ID NO:4.

[0049] AmplificationTaSIT2 The primers for the gene are as follows:

[0050] TaSIT2 -cDNA-F:5'-ATGAAGCTCACGATGCTCTTCC-3' (SEQ ID NO:3);

[0051] TaSIT2 -cDNA-R:5'-TCATCTTCCACCTGAGATGTAAGATAC-3' (SEQ ID NO:4).

[0052] Example 2

[0053] This embodiment provides the application of wheat lectin receptor kinase TaSIT2 in improving wheat rust-resistant varieties, created using CRISPR / Cas9 gene editing technology. TaSIT2 Gene editing of plants involves the following steps:

[0054] filter TaSIT2 The nucleotide sequences of the two specific targets of the gene, namely target-1 and target-2, are as follows:

[0055] target-1: 5'-CCTGGTGCTTACCAGCCGCA-3' (SEQ ID NO: 5);

[0056] target-2: 5'-CGTTCTACCCCACCCCGCTG-3' (SEQ ID NO: 6);

[0057] Based on the screened targets, forward and reverse primers were designed, and the designed primers were synthesized to synthesize target-1 primer and target-2 primer, respectively.

[0058] Target primer - target-1:

[0059] TaSIT2-SgRNA1-F:5'-CCTGGTGCTTACCAGCCGCA-3' (SEQ ID NO:7);

[0060] TaSIT2-SgRNA1-R:5'-TGCGGCTGGTAAGCACCAGG-3' (SEQ ID NO:8).

[0061] Target primer-target-2:

[0062] TaSIT2-SgRNA2-F:5'-CGTTCTACCCCACCCCGCTG-3' (SEQ ID NO:9);

[0063] TaSIT2-SgRNA2-R:5'-CAGCGGGGTGGGGTAGAACG-3' (SEQ ID NO:10).

[0064] The intermediate vector pETRN:gRNA4 was digested with BtgZI restriction enzyme. The digested vector was ligated with the target-1 primer using T4 ligase, and the ligation product was transformed into *E. coli*. Single colonies were picked and tested; positive colonies were detected. After successful sequencing, single colonies were cultured and the plasmid sgRNA-target-1 was extracted. sgRNA-target-1 was digested with BsaI, and the digested vector was ligated with target-2 using T4 ligase. The ligation product was transformed into *E. coli*, and single colonies were picked and tested; positive colonies were detected. After successful sequencing, single colonies were cultured and the plasmid was extracted. The sgRNA-target-1-target-2 was ligated with the final vector PCL4B-Cas9 using the LR reaction. Positive colonies were picked and cultured to extract the plasmid. The prepared plasmid was introduced into Fielder wheat using *Agrobacterium*-mediated transformation. TaSIT2 Gene-edited plants. This invention does not specifically limit the restriction enzymes and ligases used; any conventional enzymes well-known to those skilled in the art can be employed. The intermediate vector pETRN:gRNA4 and the final vector PCL4B-Cas9 used in this invention were provided by the Plant Immunology Team of the College of Plant Protection, Northwest A&F University, and are disclosed in the following literature: [He J, Li Q, Liang C, Wang N, Wang J, Jiang C, Kang Z, Tang C, Wang X. 2025. Transcriptional and post-translational regulation of wheat TaWRKY40 orchestrates ROS-mediated plant resistance against stripe rust. Molecular Plant, 18(10): 1688-1705.].

[0065] Example 3

[0066] The application of wheat lectin receptor kinase TaSIT2 in the improvement of wheat rust-resistant varieties provided in this embodiment, namely, the verification method for the application of wheat lectin receptor kinase TaSIT2 in the breeding and improvement of wheat rust-resistant varieties, specifically includes:

[0067] S101: Acquires wheat lectin receptor kinase TaSIT2 Gene-edited wheat, for the obtainedTaSIT2 Gene-edited wheat was sequenced using the Hi-TOM gene editing technique.

[0068] S102: Compared to the T2 generation TaSIT2 Gene-edited plants were inoculated with major prevalent races of stripe rust to identify their resistance to these races and determine their overall resistance. TaSIT2 The stripe rust resistance of gene-edited wheat varieties.

[0069] Detection using Hi-TOM gene editing sequencing technology Figure 2 The results of gene editing in gene-edited wheat plants are as follows: TaSIT2 As shown, Fielder-3B represents wild-type Fielder wheat plants in... TaSIT2 The nucleotide sequence of the gene (i.e., chromosome 3B). TaSIT2 KO-3B indicates TaSIT2 Gene-edited wheat plants in TaSIT2 The nucleotide sequence of the gene location (i.e., chromosome 3B).

[0070] Select T2 generation Figure 3 Gene-edited wheat lines L2, L4, L5, and L6 were inoculated with stripe rust fungus CYR32 at the two-leaf stage when they reached the "two-leaf-one-heart" stage. Wild wheat (Fielder) served as the control group and was also inoculated with stripe rust fungus CYR32. The growth of spore masses on wheat leaves was observed 14 days after inoculation, and the results are as follows: TaSIT2 As shown: TaSIT2 Gene-edited plants TaSIT2 KO#L2-1-1、 TaSIT2 KO#L2-2-1、 TaSIT2 KO#L4-2-6、 TaSIT2 KO#L4-3-1、 TaSIT2 KO#L5-1-7、 TaSIT2 KO#L6-2-3、 TaSIT2 The number of spore masses in KO#L6-1-6 was significantly less than that in the control group, wild wheat (Fielder), indicating that... TaSIT2 Gene-edited plants enhance wheat's resistance to stripe rust, suggesting... ​ Genes play a negative regulatory role in wheat resistance to stripe rust.

[0071] In summary, the application of wheat lectin receptor kinase TaSIT2 in improving wheat rust-resistant varieties provided in this embodiment of the invention utilizes CRISPR / Cas9 gene editing technology to construct a gene editing vector for wheat lectin receptor kinase TaSIT2. This vector is then delivered to the recipient wheat Fielder via Agrobacterium-mediated wheat genetic transformation. The resulting gene-edited plants were identified as positive plants. Four lines (L2, L4, L5, and L6) from the T2 generation of these positive plants were inoculated with the prevalent race CYR32. After 14 days, observation revealed that the gene-edited plants of wheat lectin receptor kinase TaSIT2 exhibited reduced sporulation compared to Fielder plants, demonstrating enhanced resistance.

[0072] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A wheat lectin-like receptor kinase TaSIT2 and a gene encoding the same TaSIT2 for use in increasing resistance to stripe rust in wheat; the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown as SEQ ID NO: 2; The application comprises: Knocking out genes in recipient wheat to increase resistance to stripe rust TaSIT2 .

2. Use according to claim 1, characterized in that, The method for knocking out comprises a CRISPR / Cas9 gene editing technology.

3. Use according to claim 1 or 2, characterized in that, Nucleic acid molecules for knocking out TaSIT2 The nucleic acid molecules for knocking out a gene comprise a first specific target and a second specific target, the nucleotide sequences of which are shown in SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

4. Use according to claim 1 or 2, characterized in that, knockout of the TaSIT2 gene using a gene editing vector comprising an initial vector and a nucleic acid molecule inserted into the initial vector, the nucleic acid molecule comprising a first specific target and a second specific target, the nucleotide sequences of the first specific target and the second specific target being shown in SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

5. Use according to claim 1 or 2, characterized in that, Knock out the TaSIT2 The gene uses engineered bacteria, which include gene editing vectors; The gene editing vector comprises an initial vector and a nucleic acid molecule inserted into the initial vector; The nucleic acid molecule comprises a first specific target and a second specific target, and the nucleotide sequences of the first specific target and the second specific target are respectively shown as SEQ ID NO: 5 and SEQ ID NO:

6.

6. A method of breeding wheat resistant to stripe rust, characterized in that, comprising the step of knocking out a gene in a recipient wheat TaSIT2 to obtain a stripe rust resistant wheat; The TaSIT2 The gene encodes a wheat lectin-like receptor kinase TaSIT2, the amino acid sequence of which is shown as SEQ ID NO:

2.

7. The method of claim 6, wherein, The knockout receptor wheat TaSIT2 Genetic methods include: using Agrobacterium-mediated genetic transformation to generate genes containing knockout cells. TaSIT2 The gene-editing vector containing the nucleic acid molecule of the gene was introduced into recipient wheat; TaSIT2 The nucleotide sequence of the open reading frame of the gene is shown in SEQ ID NO:

1.

8. A wheat material having resistance to stripe rust, characterized in that, compared to wild type wheat, the gene is knocked out in the wheat material; the TaSIT2 gene encodes a wheat lectin-like receptor kinase TaSIT2, the amino acid sequence of which is shown as SEQ ID NO:

2. TaSIT2 gene encodes a wheat lectin-like receptor kinase TaSIT2, the amino acid sequence of which is shown as SEQ ID NO: 2.

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