Effector protein Pst4121 of stripe rust fungus and its encoding gene and application

Abstract

The present invention belongs to the field of genetic engineering technology, and relates to a stripe rust effector protein Pst4121 and its encoding gene and application. The present invention screens and obtains the stripe rust effector protein Pst4121 based on the transcriptome sequencing of wheat stripe rust haustoria. The effector protein Pst4121 is expressed in large quantities in the early stage of stripe rust infection and reaches a peak at 48h. The present invention can transiently express the effector protein Pst4121 in tobacco through a PVX vector and identify the function of the effector protein. Genes are obtained by Agrobacterium-mediated genetic transformation. Pst4121 The invention enriches the knowledge of the wheat stripe rust effector protein by those skilled in the art, lays a theoretical foundation for clarifying the pathogenic mechanism of wheat stripe rust, and provides a theoretical basis for the subsequent creation of disease-resistant wheat varieties.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering, and relates to a stripe rust effector protein Pst4121 and a coding gene and application thereof. Background Art

[0002] Wheat stripe rust, caused by wheat stripe rust, is a disease that seriously threatens wheat production. Therefore, improving wheat stripe rust prevention and control measures and ensuring food safety production have become urgent needs that need to be urgently addressed in agricultural production. At present, the most effective and environmentally friendly prevention and control measure is to cultivate disease-resistant varieties. However, with the rapid mutation of wheat stripe rust, more and more disease-resistant varieties have become ineffective, making the technical difficulty of variety improvement increasingly greater and the waste increasing. The factors that cause the "loss" of wheat varieties' resistance to stripe rust mainly include pathogenic variation of wheat stripe rust, loss of wheat varieties' own resistance, and environmental factors. The main pathways for pathogenic variation of wheat stripe rust are mutation, heterokaryotic effect, and sexual reproduction. Among them, sexual reproduction is an important factor that accelerates its mutation rate. Stripe rust can reorganize and separate genetic material through sexual reproduction, which provides practical support for the study of sexual reproduction in wheat pathogenicity mutation. It is still unknown whether different specialized types of stripe rust can reproduce sexually. Research is still in progress in order to further improve the exploration of the pathogenicity of stripe rust.

[0003] As the systematic theory of plant immunity gradually takes shape, it provides a solid theoretical basis for plants to resist the invasion of pathogens. Plant immunity theory shows that when pathogens infect plants, plants will quickly initiate a defense response. There are two types of receptors on the surface and inside the plant that can recognize pathogens, send signals and stimulate immunity to protect themselves. In order to avoid recognition, pathogens evolve effector proteins to interfere with the host and help them infect. The receptors responsible for recognizing pathogens on the cell surface are called pattern recognition receptors, which are composed of single-transmembrane receptor-like kinases and receptor-like proteins. They stimulate immune responses by recognizing pathogens and their own molecules related to infection damage. This response is called PTI response. There is also a type of receptor-like protein containing nucleotide binding domains and leucine-rich repeat regions in the cell, which can recognize effector proteins entering plant cells and activate immune responses. This type of response is called ETI response. However, in order to achieve the purpose of infection, pathogens will evolve against the plant immune system and evade the recognition of plant disease resistance genes through the mutation of effector genes. Afterwards, in order to cope with the increasingly toxic and unrecognizable pathogens, the host plants evolved corresponding disease-resistant genes, starting a new round of tug-of-war.

[0004] However, there are still many mechanisms to be discovered in the field of plant immunity and effector proteins. As biological molecules with secretory functions that promote self-infection by regulating the host immune response, effector proteins play an important role in the interaction between plants and pathogens. Therefore, in-depth research on pathogen effector proteins is of great significance for the pathogenic mechanism of the interaction between pathogens and host plants and the molecular mechanism of pathogen evolution. Through further in-depth research on these research results, we will lay the foundation for creating more varieties with stable disease resistance and make solid contributions to ensuring food security for the people of the world. Summary of the invention

[0005] The purpose of the present invention is to provide the application of stripe rust effector proteins and their encoding genes in plant breeding or creation, aiming to further enrich and understand the wheat stripe rust effector proteins for technicians in this field, and provide theoretical cognition and gene selection for the breeding and creation of excellent stripe rust-resistant germplasm materials.

[0006] In order to fully understand the technical solution of the present invention without objection, it is necessary to add that the coding gene of the stripe rust effector protein Pst4121 of the present invention is represented by italic font “ Pst4121 " indicates that the stripe rust effector protein Pst4121 is represented by the non-italic font "Pst4121". Of course, a person skilled in the art can clearly and completely understand the meaning and expression of the relevant genes and their encoded proteins based on the description of the present invention.

[0007] On the one hand, the present invention relates to a stripe rust effector protein Pst4121, which has an amino acid sequence as shown in SEQ ID NO: 1, and the N-terminal 1 to 20 amino acids are a signal peptide sequence.

[0008] SEQ ID NO: 1 is specifically as follows,

[0009] MNTYHIYLFLSLALYPGAFSLLTGFDPKNLPYKSDGAAGQTGYNRCGTKATKDSQCQNLFINSAEDFCVFAPPTVLSIGEAERIAVSYCSKNGHGTRLIPPGTFRTMHYVRTQHYIQITALGDFTKINVPAGDEGGE LDPSGADGHGNPIGGLVFGERGQFNQWTEFLAHDELSIRACFNTDQAYRYCQHIYDLEGSRWNMPGNYDGAGFDQCEGDDVPHPMGEYRRPDGSIYTWQRGHGPQPPAGQPGKIKSCKSVRAPGLNYPRSAKRNSIS.

[0010] In another aspect, the present invention relates to Pst4121A gene, which comprises the nucleotide sequence of the stripe rust effector protein Pst4121.

[0011] Furthermore, in the gene provided by the present invention Pst4121 the nucleotide is as shown in SEQ ID NO: 2.

[0012] SEQ ID NO: 2 is specifically as follows:

[0013] ATGAACACATACCACATTTATCTGTTCCTGTCGCTGGCTCTCTATCCAGGTGCCTTCTCCCTCCTAACCGGATTTGATCCCAAAAACCTACCCTACAAAAGTGACGGCGCCGCCGGTCAAACAGGTTATAATCGATGTGGCACCAAAGCTACTAAAGACAGCCAATGCCAGAATCTTTTTATCAACTCCGCCGAGGACTTCTGCGTGTTTGCCCCCCCCACGGTATTATCCATTGGTGAGGCAGAGCGTATTGCTGTTTCTTACTGTTCGAAAAACGGCCATGGTACTCGTCTGATTCCCCCAGGAACCTTCAGAACGATGCACTACGTTCGCACGCAACACTACATCCAAATAACCGCCCTTGGTGACTTCACCAAAATTAACGTCCCGGCGGGAGATGAAGGTGGCGAATTAGATCCCTCCGGTGCTGACGGTCACGGTAACCCCATCGGTGGACTGGTCTTTGGCGAGCGAGGCCAATTCAACCAATGGACCGAATTCCTTGCTCACGATGAGCTGTCCATCAGAGCATGCTTCAATACCGACCAAGCTTACAGGTACTGCCAACACATTTATGATCTGGAGGGCAGTCGCTGGAACATGCCTGGTAACTACGACGGTGCTGGGTTCGATCAATGTGAGGGTGACGATGTACCCCATCCCATGGGTGAATATCGCCGACCCGACGGTTCTATCTACACTTGGCAGCGAGGTCATGGTCCCCAACCCCCAGCCGGTCAACCGGGCAAAATTAAATCATGCAAGAGCGTGCGGGCCCCAGGCTTGAACTATCCCCGTTCCGCCAAACGTAACTCAATTTCTTAG.

[0014] Further, in the present invention provided Pst4121 In genes, silencing Pst4121 Gene reduces wheat's resistance to stripe rust.

[0015] On the other hand, the present invention relates to the stripe rust effector protein Pst4121 or the Pst4121 Application of genes in the control of stripe rust.

[0016] In another aspect, the present invention relates to an expression vector comprising Pst4121 Gene.

[0017] On the other hand, the present invention relates to a method for breeding wheat varieties resistant to stripe rust, characterized in that the expression vector is transferred into wheat to overexpress the Pst4121 Gene.

[0018] In another aspect, the present invention relates to a method for instantaneously silencing said Pst4121 Gene vector.

[0019] Furthermore, the vector provided by the present invention comprises a silencing fragment having a nucleotide sequence as shown in SEQ ID NO:3.

[0020] On the other hand, the present invention relates to a method for genetically improving wheat resistance to stripe rust, which comprises transferring the vector into wheat.

[0021] Those skilled in the art can easily use known methods, such as directed evolution and point mutation methods, to mutate the nucleotide sequence encoding the stripe rust effector protein Pst4121 of the present invention. Pst4121 As long as the nucleotides with 75% or higher identity to the nucleotide sequence of the present invention encode the stripe rust effector protein Pst4121 and have the same function, they are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention.

[0022] The term "identity" as used herein refers to sequence similarity to a natural nucleic acid sequence. "Identity" includes nucleotide sequences that have 75% or more, or 85% or more, or 90% or more, or 95% or more identity to the nucleotide sequence of the present invention encoding the protein consisting of the amino acid sequence shown in SEQ ID NO: 1. Identity can be evaluated by the naked eye or by computer software. Using computer software, the identity between two or more sequences can be expressed as a percentage (%), which can be used to evaluate the identity between related sequences.

[0023] The above-mentioned 75% or more identity may be 80%, 85%, 90% or 95% or more identity.

[0024] Those skilled in the art can easily use known methods, such as gene Pst4121 Gene silencing cassette Pst4121 The product, the expression cassette (gene) containing the nucleic acid molecule encoding the stripe rust effector protein Pst4121 Pst4121 Expression cassette) refers to DNA that can express stripe rust effector protein Pst4121 in host cells, which can include not only the promoter gene Pst4121 The promoter of transcription may also include the terminator gene Pst4121 The expression cassette may further include an enhancer sequence. The promoters that can be used in the present invention include, but are not limited to, constitutive promoters; tissue, organ and development-specific promoters and inducible promoters. Suitable transcription terminators include, but are not limited to, Agrobacterium nopaline synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator and nopaline and octopine synthase terminators.

[0025] Existing expression vectors can be used to construct Pst4121Recombinant vector of expression cassette. The plant expression vector includes binary Agrobacterium vector and vector that can be used for plant microprojectile bombardment, etc. Such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1305, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA Company), etc. The plant expression vector may also contain the 3' non-translated region of the foreign gene, i.e., a polyadenylic acid signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylic acid signal can guide the addition of polyadenylic acid to the 3' end of the mRNA precursor, such as the Agrobacterium crown gall induction (Ti) plasmid gene (such as the nopaline synthase gene Nos), the 3' non-translated region of the plant gene (such as the soybean storage protein gene) has similar functions. When using the gene construction plant expression vector of the present invention, enhancers can also be used, including translation enhancers or transcription enhancers. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be identical to the reading frame of the coding sequence to ensure the correct translation of the entire sequence. The source of the translation control signal and the start codon is extensive and can be natural or synthetic. The translation initiation region can come from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed, such as adding genes that can be expressed in plants and encode enzymes or luminescent compounds that can produce color changes (GUS gene, luciferase gene, etc.), antibiotic marker genes (such as nptII gene that confers resistance to kanamycin and related antibiotics, bar gene that confers resistance to the herbicide phosphinothricin, hph gene that confers resistance to the antibiotic hygromycin, dhfr gene that confers resistance to methotrexate, EPSPS gene that confers resistance to glyphosate), or chemical resistance marker genes (such as herbicide resistance genes), mannose-6-phosphate isomerase gene that provides the ability to metabolize mannose. Considering the safety of transgenic plants, no selective marker genes can be added, and transformed plants can be directly screened by adversity.

[0026] The above-mentioned vector or expression vector can be a plasmid, cosmid, phage or virus vector.

[0027] The microorganisms may be yeast, bacteria, algae or fungi, such as Agrobacterium.

[0028] The above transgenic plant cell lines do not include propagation materials.

[0029] The above disease resistance is resistance to wheat stripe rust.

[0030] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0031] (1) The wheat stripe rust effector protein Pst4121 provided by the present invention is overexpressed in tobacco and wheat, and it is clear that Pst4121 can inhibit PCD reactions and wheat defense reactions PTI and ETI reactions.

[0032] (2) The wheat stripe rust effector protein Pst4121 provided by the present invention can be used to create a plant with silenced stripe rust effector protein Pst4121 using host wheat, and it can be clearly seen that the stripe rust susceptibility is weakened. Therefore, the effector protein Pst4121 can be used to create rust-resistant wheat lines, providing excellent materials for the cultivation of wheat stripe rust-resistant varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0034] Figure 1 Puccinia striiformis effector protein gene Pst4121 Analysis of expression levels at different times after stripe rust race CYR31 infected wheat. Figure 1 The data are the mean ± SEM of three biological replicates.

[0035] Figure 2 Puccinia striiformis effector protein gene Pst4121 Fluorescence localization status diagram. Figure 2 GFP in the column indicates GFP protein, and Pst4121-GFP indicates the fusion protein of Pst4121 and GFP.

[0036] Figure 3 Figure 3 shows the inhibition of Bax- and DC3000-induced PCD by Pst4121. Figure 3 Middle: A and C are photos of the same leaf before and after using the decolorizing solution; B is a description of the specific injection of tobacco. 7 and 10 represent the soybean phytophthora effector protein Avr1b as a positive control, 6 and 11 represent the GFP empty vector as a negative control, and 8 and 9 show that Pst4121 can inhibit the PCD reaction induced by Bax and DC3000, respectively.

[0037] Figure 4 To use RNAi technology Pst4121 Phenotypic analysis of CYR31 inoculated after silencing, where L5, L11 and L12 represent three different strains. DETAILED DESCRIPTION

[0038] The technical scheme of the present invention is described below in conjunction with the examples, but the present invention is not limited to the following examples. The experimental methods and detection methods described in each example are conventional methods unless otherwise specified; the reagents and materials are available on the market unless otherwise specified, and the % in the following examples are all mass percentages unless otherwise specified.

[0039] Example 1

[0040] This example provides the stripe rust effector protein gene after the stripe rust race CYR31 infects wheat. Pst4121 Expression analysis at different time points.

[0041] 1. Acquisition of Stripe Rust Real-time Fluorescence Quantitative PCR Samples

[0042] Wheat seeds (water source 11) were planted in a 10 cm diameter pot and cultured under a 16 / 8 h light / dark cycle and a temperature of 16 ℃. When the wheat seedlings grew to the one-leaf-one-heart stage, fresh wheat stripe rust summer spores were diluted with electronic fluoride liquid, and 10 μL of the diluted spore solution was evenly smeared and inoculated on the wheat leaves with a pipette. The inoculated wheat seedlings were kept in the dark at 15 ℃ for 36 h and then continued to be cultured under normal light conditions. After the wheat leaves were inoculated, they were sampled at 6 h, 12 h, 18 h, 24 h, 48 h, 72 h, and 120 h as real-time fluorescence quantitative PCR samples. Each group of samples selected leaves from at least three different wheat seedlings, and the middle area of ​​the inoculated leaves was mainly collected. After sampling, they were immediately stored at -80 ℃ for later use.

[0043] 2. Preparation of Stripe Rust Template

[0044] The collected samples were used to prepare cDNA templates for real-time fluorescence quantitative detection of stripe rust using an RNA extraction kit and a vazyme reverse transcription kit (Beijing Huayueyang Biotechnology Co., Ltd.) according to the operating instructions.

[0045] 3. Real-time fluorescence quantitative PCR detection

[0046] The wheat stripe rust elongation factor gene PstEF was used as an internal reference gene and the gene expression was analyzed by relative quantitative method.

[0047] The sequences of the internal reference gene primers are:

[0048] PstEF-F: TTCGCCGTCCGTGATATGAACAA;

[0049] PstEF-R: ATGCGTATCATGGTGGTGGAGTGA.

[0050] Pst4121-specific primers were designed using Primer Premier 5.0 software. The sequences of Pst4121-specific primers are as follows:

[0051] DL-4121-F:ATCCCAAAAACCTACCCTACA;

[0052] DL-4121-R:GGCATTGGCTGTCTTTAGTAG.

[0053] UltraSYBR One Step RT-qPCR Kit (Kangwei Century Biotechnology Co., Ltd.) was used according to the instructions, and the relevant detection was completed using the Bio-Rad iQ5 real-time fluorescence quantitative PCR instrument. The relative expression of different genes was based on 2 -ΔΔCt The expression level of PstEF was used as a reference for calculation.

[0054] The results are as follows Figure 1 As shown in the figure, in the early stage of wheat stripe rust infection, the expression level of effector protein Pst4121 was high, reaching a peak at 48h, with an expression level of about 25 times more, and suddenly dropped at 72h, with almost no expression. This shows that Pst4121 plays an important role in the early stage of wheat stripe rust infection.

[0055] Example 2

[0056] This example provides a localization test of the effector protein Pst4121 in cells.

[0057] 1. Construction of subcellular localization plasmid of stripe rust effector protein gene Pst4121

[0058] Primer Premier 5.0 software was used to design primers based on the Pst4121 sequence without the signal peptide. Pst4121 -pBIN-F, Pst4121 -pBIN-R. The primer sequences are as follows:

[0059] Pst4121- pBIN-F:GACGAGCTGTACAAGGGTACCATGCTCC

[0060] GGATTTGAT;

[0061] Pst4121- pBIN-R:TCTAGTTCATCTAGAGGATCCAGAAATTGAGTTACGTTTG.

[0062] Using wheat stripe rust cDNA as template, 2×Taq MasterMix PCR mixture (Kangwei Century Biotechnology Co., Ltd.) and full-length gene primers were used for PCR amplification. The reaction system is:

[0063]

[0064] The PCR instrument program was as follows: 95°C for 5 min pre-denaturation, 95°C for 15 s denaturation, 58°C for 30 s annealing, 72°C for 1 min extension, 40 cycles in total, and 72°C for 10 min additional extension. After the PCR program was completed, the PCR product was tested by agarose gel recovery gel, and the bands that matched the size of the target gene were recovered using the omega gel recovery kit, and the operation was performed according to the instructions (Omega Bio-Tek).

[0065] The recovered glue Pst4121 The gene fragment was connected to the vector PBIN-GFP and transformed into the competent E. coli DH5α (Shanghai Weidi Biotechnology Co., Ltd.) to screen the positive clones. The positive clones were then sent to Beijing Qingke Biotechnology Co., Ltd. for plasmid sequencing. The plasmids with correct sequencing results were retained for the next experiment. The plasmids were labeled as: PBIN- Pst4121 .

[0066] 2. Transformation of recombinant vector into Agrobacterium competent cells GV3101

[0067] (1) Thaw the Agrobacterium competent cells GV3101 (Shanghai Weidi Biotechnology Co., Ltd.) (place on ice) and pre-cool the electroporation cuvette;

[0068] (2) Add 2 μL pBIN- Pst4121 Slowly pipette and mix well;

[0069] (3) Turn on the electric shock device in advance, set the voltage to 2500 mV, and the shock time to 5 ms;

[0070] (4) Preheat LB culture medium (28°C), add 1.5 mL of culture medium into the electroporation cup, and aspirate the mixed solution;

[0071] (5) Resuscitation of competent cells; culture in a constant temperature shaker (28°C, 200 rpm) for 2.5 h;

[0072] (6) Centrifuge the bacterial solution at 5000 rpm at room temperature, spread 150 μL of the bacterial solution on LB solid medium (rifampicin and kanamycin at a ratio of 1:5000), and culture at 28°C in the dark (to prevent rifampicin from becoming ineffective) for about 48 h;

[0073] (7) Select positive clones for sequencing and keep the bacterial solution for later use;

[0074] 3. Injection of Tobacco with Agrobacterium Containing Recombinants

[0075] (1) The above bacterial solution was placed in a constant temperature shaker at 28°C and 200 rpm for 24 h;

[0076] (2) Centrifuge at 6000 rpm for 2 min, collect the cells, resuspend the cells in 10 mM MgCl2 and wash, repeat three times; dilute the cells with 10 mM MgCl2 to an OD of 600 = about 0.6;

[0077] (3) The above bacterial solution was infecting tobacco cells and cultured at 25°C, 16 h light / 8 h dark conditions for about 36 h. The fluorescence was observed and photographed using an OLYMPUS FV3000 Confocal Laser Scanning Microscope.

[0078] Example 3

[0079] This example provides a validation experiment that Pst4121 inhibits Bax and DC3000-induced cell necrosis.

[0080] 1. Vector Construction

[0081] To remove the signal peptide Pst-4121 Primer design Pst4121 - pGR107-F and Pst4121 - pGR107-R, primers are as follows:

[0082] Pst4121- pGR107-F:CACCAGCTAGCATCGATTCCCGGGATGCTCCTAACCGGATTTGAT;

[0083] Pst4121 -pGR107-R:GGAACATCGTATGGGTACGCGGGTTACGTTTGGCGGAA.

[0084] 2. Gene Pst-4121 Connected to PVX expression vector pGR107. Recombinant vector was transformed into Agrobacterium GV3101. Agrobacterium containing recombinant BAX and Pseudomonas syringae DC3000 were used to infect tobacco, the injection site was marked, and the tobacco was cultured at 25°C, 16 h light / 8 h dark for about 4 days, and the tobacco status was observed. The vector construction method, Agrobacterium transformation and tobacco injection method were the same as in Example 2.

[0085] 3. Decolorize the tobacco with a fixative (glacial acetic acid: anhydrous ethanol) and take photos. The test results are as follows: Figure 3 The above test results show that Pst4121 can inhibit cell necrosis induced by Bax and DC3000.

[0086] Example 4

[0087] Pst4121 Verification test of the conservativeness of gene-silenced transgenic plants to wheat stripe rust.

[0088] 1. Construction of transgenic vector

[0089] The ORF region of the stripe rust effector protein Pst4121 gene sequence was compared with the homologous genes of wheat and wheat stripe rust, and the Pst4121 Specific fragment sequences are used for transgenic vector construction. Pst4121 The nucleotide sequence of the gene silencing fragment (as shown in SEQ ID NO: 3) was inserted into two positions of the PC336 vector by homologous recombination (one fragment was inserted in the forward direction and the other was inserted in the reverse complementary direction, so as to form a hairpin structure), and the PC336-4121 plasmid was obtained.

[0090] The primer sequence containing the attB site is:

[0091] attB-4121-F:

[0092] GGGGACAAGTTTGTACAAAAAAGCAGGCTCGATGTGGCACCAAAGCTAC;

[0093] attB-4121-R:

[0094] GGGGACCACTTTGTACAAGAAAGCTGGGTGAAGTCACCAAGGGGCGGTTA.

[0095] 2. Transformation of Agrobacterium

[0096] Take the EHA105 competent cells stored at -80 ℃ and melt them on ice, add the correctly sequenced PC336-4121 plasmid, ice bath for 30 min, liquid nitrogen for 2 min, 37 ℃ water bath for 3 min, ice bath for 5 min, add appropriate amount of liquid LB medium, and shake culture for 3 h; take 150 μL of the transformation bacterial liquid and spread it on a solid plate (rifampicin and Kana antibiotics), invert and culture in the dark until positive transformants grow.

[0097] Using Agrobacterium-mediated wheat transformation technology, the recombinant silencing vector PC336-4121 was transferred into the wheat variety Fielder material to obtain RNAi silenced plants. DNA testing was performed on the obtained plants, and positive plants were retained for subsequent verification.

[0098] 3. Phenotypic Identification of Transgenic Plants

[0099] Three T3 transgenic homozygous strains were selected for disease resistance testing and inoculated with wheat stripe rust CYR31 in the same manner as in Example 1. Phenotypic identification was performed after sporulation of wheat leaves, and photographs were taken for record.

[0100] The results are as follows Figure 4 As shown, the spore production of stripe rust races on the silenced plants inoculated with wheat stripe rust CYR31 was significantly reduced, indicating that the effector protein Pst4121 proposed in the present invention has the function of promoting the stripe rust to infect wheat during the process of the stripe rust infecting wheat.

[0101] In summary, the stripe rust effector protein Pst4121 proposed in the present invention plays a key role in the process of stripe rust infection of wheat. By constructing transgenic RNAi plants to silence the expression of this gene, a significant disease resistance response to wheat stripe rust is produced. This gene has a reference significance for the creation of broad-spectrum disease-resistant materials.

[0102] As described above, the basic principle, main features and advantages of the present invention are well described. The above embodiments and descriptions are only descriptions of the preferred implementation modes of the present invention, and the present invention is not limited by the above embodiments. Without departing from the spirit and scope of the present invention, various changes and improvements made by ordinary technicians in this field to the technical solution of the present invention should fall within the protection scope determined by the present invention.

Claims

1. Pst4121 The application of the gene in the prevention and treatment of stripe rust is characterized in that: Said Pst4121 The nucleotide sequence of the gene is shown in SEQ ID NO: 2, silencing the Pst4121 Genes improve wheat resistance to stripe rust.

2. The use according to claim 1, characterized in that: The fragment with the nucleotide sequence shown in SEQ ID NO: 3 is said Pst4121 Gene-silencing fragments of genes.

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