Application of wheat lectin receptor-like kinase TaSIT2 and coding gene TaSIT2 thereof in improving stripe rust resistance of crops

Abstract

The invention belongs to the technical field of plant genetic engineering and crop breeding, and particularly relates to application of wheat lectin receptor-like kinase TaSIT2 and a coding gene TaSIT2 thereof in improving stripe rust resistance of crops. A wheat lectin receptor-like kinase gene TaSIT2 is analyzed and screened by using a reverse genetics method, a TaSIT2 gene editing wheat plant is obtained by using an agrobacterium-mediated genetic transformation method through a gene editing technology, the resistance of the transgenic plant to wheat stripe rust physiological races is remarkably enhanced, and the wheat stripe rust physiological races can be obtained. It is shown that knockout of the TaSIT2 gene enhances the resistance of wheat to stripe rust, it is determined that the wheat lectin receptor-like kinase TaSIT2 gene plays a negative regulation role in wheat stripe rust resistance, and a new technical thought and gene resources are provided for wheat stripe rust resistance variety cultivation from the perspective of molecular biology. And more paths are provided for cultivating lasting broad-spectrum disease-resistant varieties.

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 application further provides a transformant comprising at least one of an engineered bacterium, a transgenic crop cell and a transgenic crop tissue, wherein the transformant comprises the nucleic acid molecule or the gene editing vector.

[0013] The application further provides an application of the nucleic acid molecule or the gene editing vector or the transformant in improving the resistance of crops to stripe rust and / or cultivating a stripe rust-resistant crop variety.

[0014] Preferably, the crop comprises wheat.

[0015] The application further provides a method for cultivating a stripe rust-resistant wheat, comprising the following steps: reducing the expression amount of a wheat lectin-like receptor kinase TaSIT2 gene in a recipient wheat or inhibiting the expression of the wheat lectin-like receptor kinase TaSIT2 gene in the recipient wheat, to obtain the stripe rust-resistant wheat. TaSIT2 The gene encodes the wheat lectin-like receptor kinase TaSIT2, and the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown as SEQ ID NO: 2. The gene encodes the wheat lectin-like receptor kinase TaSIT2, and the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown as SEQ ID NO: 2. TaSIT2 The gene encodes the wheat lectin-like receptor kinase TaSIT2, and the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown as SEQ ID NO: 2.

[0016] Preferably, the method for reducing the expression amount of the wheat lectin-like receptor kinase TaSIT2 gene in the recipient wheat or inhibiting the expression of the wheat lectin-like receptor kinase TaSIT2 gene in the recipient wheat comprises the following steps: introducing a gene editing vector containing a nucleic acid molecule for knocking out and / or knocking down the wheat lectin-like receptor kinase TaSIT2 gene into the recipient wheat by an Agrobacterium-mediated genetic transformation method. TaSIT2 The open reading frame of the gene has a nucleotide sequence as shown in SEQ ID NO: 1. TaSIT2 TaSIT2 Beneficial effects: The application provides a wheat lectin-like receptor kinase TaSIT2 and an encoding gene thereof

[0017] In the application of improving the resistance of crops to stripe rust, the application first discloses the role of the wheat lectin-like receptor kinase TaSIT2 in the resistance to stripe rust, and creates a high-resistance germplasm resource by using a transgenic technology, thereby providing a solution to the problems of a lack of wheat stripe rust resistance resources and difficulty in preventing and controlling wheat stripe rust. Specifically, the application clarifies the function of the wheat lectin-like receptor kinase TaSIT2 in the interaction between wheat and stripe rust, and uses a CRISPR / Cas9 gene editing technology to knock out the wheat lectin-like receptor kinase TaSIT2 gene in a wheat plant in vivo, thereby enhancing the resistance of the wheat to stripe rust pathogens. TaSIT2 TaSIT2 TaSIT2 ​​​The gene edited wheat variety shows resistance to the main epidemic race of Puccinia striiformis. The application provides a new technical idea for cultivating wheat varieties resistant to stripe rust from the perspective of molecular biology, and provides a gene resource for wheat resistance to stripe rust, and provides more paths for cultivating varieties with persistent and broad-spectrum resistance. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the drawings needed in the embodiments will be briefly introduced as follows.

[0019] Figure 1 A flow chart of a verification method for the application of the wheat lectin-like receptor kinase TaSIT2 in cultivating and improving wheat varieties resistant to rust; Figure 2 In Example 3 TaSIT2 A schematic diagram of gene editing of the gene edited wheat plant; Figure 3 In Example 3 TaSIT2 A phenotype result diagram of inoculating the gene edited wheat plant with the compatible race CYR32 of Puccinia striiformis, wherein “Fielder” represents a wild type Fielder wheat plant; 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 respectively represent TaSIT2 Four strains #L2, #L4, #L5 and #L6 of the gene wheat plant. DETAILED DESCRIPTION

[0020] The application provides a wheat lectin-like receptor kinase TaSIT2 and an encoding gene thereof TaSIT2 and / or application in breeding of crops resistant to stripe rust; the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown as SEQ ID NO: 2.

[0021] As an implementation form, the application provides a wheat lectin-like receptor kinase TaSIT2 TaSIT2 The nucleotide sequence of the open reading frame (ORF) of the gene is shown as SEQ ID NO: 1.

[0022] The sequences of SEQ ID NO: 1 to SEQ ID NO: 2 of the application are as follows: SEQ ID NO: 1 is specifically as follows: SEQ ID NO: 2 is specified as follows: MKLTMLFLVLFSFHLAAGDDVQVQFAFPGLAGANLTLDGNATVTPDGLLVLTSRKTNLKGHAFYPTPLRFRGSPGGQVHSFASSFVFAIVSDYTDFSAHGMALVVCPTADSLSSALPAGYLALLNVQNNGNASNHLFAVELDTTQNTDFQDINANHVGIDVNDLHSLQSYPTGYYLDDGGDFRNLTLFSREVMQVWVNYDGGTGQIDVTLAPIGVARPARPLVSATYNLSSVITEQAYIGFSSATGGINTRHYVLAWSFAMNGPAPAIDVAKLPKLPRFGPKPRSKVLEIVLPIATAVIILAVCTVVALLLLRHLRYAQLLEDWEVEFGPHRFSYKELYHATKGFKSKHLLGAGGFGKVYKGVLHKSKMEVAIKKISHESRQGMKEFIAEVVSIGRLRNRNLVQLLGYCRRKGELLLVYEYMPNGSLDKYLYGECDENNVHVLDWTKRLHIIKGVASGLLYIHEKWEKIVIHRDIKASNVLLDSEMNGRLGDFGLARLYDHGTDPQTTHVVGTMGYLAPELVRTGKASTLTDVFAFGAFLLEITCGRRPVSNGPQDSHEMLVDWVLDHFRRGCLAETVDVRMGGDYSVDEACRVLKLGLLCSHPFASTRPSMRHVMQYLDSDTPLPDLSSADMMSFSTMALLQNQGFDSYAISYPSSMGSIGTVSYISGGR.

[0023] As an embodiment, the application comprises: knocking out and / or knocking down in a recipient plant TaSIT2As another implementation form, the knockout and / or knockdown method comprises a CRISPR / Cas9 gene editing technology; more specifically, a gene editing vector is constructed by the CRISPR / Cas9 gene editing technology, and the gene editing vector is introduced into a recipient crop by an Agrobacterium-mediated genetic transformation method, so as to obtain a wheat lectin-like receptor kinase TaSIT2 gene editing crop variety. TaSIT2 The gene editing wheat variety can improve the resistance of the wheat to the stripe rust.

[0024] The present application also provides a knockout and / or knockdown TaSIT2 The nucleic acid molecule comprises a first specific target and a second specific target, and the nucleotide sequences of the amplified first specific target and second specific target are respectively shown in SEQ ID NO: 5 and SEQ ID NO: 6. As an implementation form, the first specific target and the second specific target are amplified by a target primer-target-1 and a target primer-target-2, respectively, wherein the forward primer and the reverse primer of the target primer-target-1 comprise the nucleotide sequences shown in SEQ ID NO: 7 and SEQ ID NO: 8, respectively, and the forward primer and the reverse primer of the target primer-target-2 comprise the nucleotide sequences shown in SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

[0025] The present application 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. As an implementation form, the initial vector is a Cas9 plasmid vector. The construction method of the gene editing vector is not particularly limited in the present application, and a conventional construction method in the art can be used.

[0026] The present application also provides a transformant comprising at least one of an engineering bacterium, a transgenic crop cell and a transgenic crop tissue, wherein the transformant comprises the nucleic acid molecule described in the above technical solution or the gene editing vector described in the above technical solution. The transgenic crop cell and the transgenic crop tissue are non-stable genetic cells and tissues. As an implementation form, the crop can be a gramineous crop or wheat.

[0027] 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. 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; 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.

[0028] 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.

[0029] 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.

[0030] 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.

[0031] The experimental methods and detection methods in the following embodiments are conventional methods unless otherwise specified. The reagents and materials can be purchased on the market unless otherwise specified. The index data can be measured by conventional methods unless otherwise specified.

[0032] Figure 1 A flow chart of the verification method of the application of the wheat lectin-like receptor kinase TaSIT2 in breeding and improving wheat varieties resistant to rust is given. The verification methods used for other host plants of stripe rust are the same as or slightly different from TaSIT2 the technical ones. The embodiment of the application provides a verification method of the function of the wheat lectin-like receptor kinase TaSIT2, comprising: S101, obtaining a wheat lectin-like receptor kinase TaSIT2 gene edited wheat, performing Hi-TOM gene editing sequencing on the obtained TaSIT2 gene edited wheat; S102, inoculating the T2 generation TaSIT2 gene edited plants with the main epidemic race of stripe rust, identifying the resistance of the gene edited plants to the epidemic race of stripe rust, and determining TaSIT2 the stripe rust resistance characteristics of the gene edited wheat variety.

[0033] Embodiment 1 This embodiment provides the obtaining of the coding gene sequence and the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2, and the steps are as follows: The open reading frame (ORF) sequence and the protein sequence of the wheat lectin-like receptor kinase TaSIT2 gene were obtained through the Ensembl Plants website (https: / / plants.ensembl.org / Triticum_aestivum / Info / Index). The primers for amplifying the TaSIT2 cDNA sequence of the gene were designed, and the cDNA of the wheat Fielder plant was used as a template to amplify the TaSIT2 gene and sequence it.

[0034] TaSIT2 The open reading frame (ORF) sequence of the gene is shown as SEQ ID NO: 1, and the amino acid is shown as SEQ ID NO: 2. The forward primer for amplifying the TaSIT2 cDNA sequence of the gene is shown as SEQ ID NO: 3, and the reverse primer is shown as SEQ ID NO: 4. TaSIT2 TaSIT2 The primers for amplifying the gene are as follows:

[0035] TaSIT2 TaSIT2 ​​-cDNA-F: 5'-ATGAAGCTCACGATGCTCTTCC-3' (SEQ ID NO: 3); TaSIT2 -cDNA-R: 5'-TCATCTTCCACCTGAGATGTAAGATAC-3' (SEQ ID NO: 4).

[0036] Example 2 The application of wheat lectin-like receptor kinase TaSIT2 in the improvement of wheat rust-resistant varieties is provided in this example, and a gene editing plant is created using CRISPR / Cas9 gene editing technology TaSIT2 The steps are as follows: Screening TaSIT2 The nucleotide sequences of the two specific target sites of the gene, i.e., target site 1 (target-1) and target site 2 (target-2), are as follows: target-1: 5'-CCTGGTGCTTACCAGCCGCA-3' (SEQ ID NO: 5); target-2: 5'-CGTTCTACCCCACCCCGCTG-3' (SEQ ID NO: 6); According to the target sites obtained by screening, forward primers and reverse primers are designed, and the designed primers are synthesized, target-1 primers and target-2 primers are synthesized respectively; Target primer-target-1: TaSIT2-SgRNA1-F: 5'-CCTGGTGCTTACCAGCCGCA-3' (SEQ ID NO: 7); TaSIT2-SgRNA1-R: 5'-TGCGGCTGGTAAGCACCAGG-3' (SEQ ID NO: 8).

[0037] Target primer-target-2: TaSIT2-SgRNA2-F: 5'-CGTTCTACCCCACCCCGCTG-3' (SEQ ID NO: 9); TaSIT2-SgRNA2-R: 5'-CAGCGGGGTGGGGTAGAACG-3' (SEQ ID NO: 10).

[0038] The intermediate vector pETRN:gRNA4 is single-enzymatically cut by using BtgZI endonuclease, the cut vector is connected with the target-1 primer by using T4 ligase, and the connected product is transformed into E. coli, a single colony is picked and sent for detection, positive colonies are detected, and when the sequencing result is correct, a single colony is picked and shaken to culture and extract the plasmid sgRNA-target-1; sgRNA-target-1 is cut by using BsaI, the cut vector is connected with target-2 by using T4 ligase, the obtained connection product is transformed into E. coli, a single colony is picked and sent for detection, positive colonies are detected, and when the sequencing result is correct, a single colony is picked and shaken to culture and extract the plasmid, sgRNA-target-1-target-2 is connected with the final vector PCL4B-Cas9 by using the LR reaction, a positive colony is picked and shaken to extract the plasmid, and the prepared plasmid is introduced into Fielder wheat by using agrobacterium mediation to obtain TaSIT2 gene edited plants. The endonuclease and ligase used in the present application are not particularly limited, and conventional enzymes familiar to those skilled in the art can be used. The intermediate vector pETRN:gRNA4 and the final vector PCL4B-Cas9 used in the present application are both provided by the Plant Immunity Team of the College of Plant Protection, Northwest Agriculture and Forestry 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.

[0039] Embodiment 3 The wheat lectin-like receptor kinase TaSIT2 provided in the present embodiment is applied in the improvement of wheat rust-resistant varieties, that is, a verification method for the application of the wheat lectin-like receptor kinase TaSIT2 in the cultivation and improvement of wheat rust-resistant varieties, which specifically comprises: S101: obtaining a wheat lectin-like receptor kinase TaSIT2 gene editing wheat, and the obtained TaSIT2 The gene edited wheat is subjected to Hi-TOM gene editing sequencing. S102: detecting the T2 generation of the TaSIT2 The gene edited plants are inoculated with the main epidemic race of Puccinia striiformis Westend. f. sp. tritici, the resistance of the gene edited plants to the epidemic race of Puccinia striiformis Westend. f. sp. tritici is identified, and the TaSIT2The stripe rust resistance of gene-edited wheat varieties.

[0040] 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).

[0041] 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.

[0042] 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.

[0043] Although the above embodiments have been described in detail, it should be understood that these are only some embodiments of the present application, but not all embodiments. Other embodiments can be obtained based on the above embodiments without creativity, and these embodiments all belong to the protection scope of the present application.

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 crops and / or breeding of stripe rust resistant crops; the amino acid sequence of the wheat lectin-like receptor kinase TaSIT2 is shown in SEQ ID NO:

2.

2. Use according to claim 1, characterized in that, The applications include knocking out and / or knocking down TaSIT2 genes in a recipient plant, increasing the resistance of a crop to stripe rust and / or breeding a stripe rust resistant crop variety.

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

4. A nucleic acid molecule for knocking out and / or knocking down TaSIT2 a gene, characterized in that 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.

5. A gene editing vector, characterized by, 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 transformant characterized in that, The transformant comprises at least one of an engineering bacterium, a transgenic crop cell and a transgenic crop tissue, and the transformant comprises the nucleic acid molecule of claim 4 or the gene editing vector of claim 5.

7. Use of the nucleic acid molecule of claim 4 or the gene editing vector of claim 5 or the transformant of claim 6 in improving the resistance of crops to stripe rust and / or cultivating a stripe rust resistant crop variety.

8. Use according to any one of claims 1 to 3 or transformant according to claim 6 or use according to claim 7, characterized in that, The crop comprises wheat.

9. A method of breeding wheat resistant to stripe rust, characterized in that, comprising the step of reducing the expression level of a wheat lectin-like receptor kinase TaSIT2 in the recipient wheat or inhibiting the expression of a gene in the recipient wheat TaSIT2 resistance to stripe rust; 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.

10. The method of claim 9, wherein, The reduction of wheat lectin receptor kinase TaSIT2 expression or inhibition of expression in 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.

Images