Plant disease resistance-related protein TaCNGC10 and its encoding gene and applications

Abstract

This invention belongs to the field of biotechnology, and particularly relates to the plant disease resistance-related protein TaCNGC10, its encoding gene, and its applications. This invention provides a plant disease resistance-related protein TaCNGC10, the amino acid sequence of which is shown in SEQ ID NO:2. Through functional analysis of the related protein and gene, this invention reveals its disease resistance mechanism in the interaction between plants and stripe rust, providing a solution for the breeding of stripe rust-resistant materials.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and in particular relates to the plant disease resistance-related protein TaCNGC10, its encoding gene, and its applications. Background Technology

[0002] Stripe rust fungi are obligate parasitic organisms, meaning their growth and reproduction are entirely dependent on host plants, from which they must obtain water and nutrients. Different stripe rust races have varying effects on their host plants. Since stripe rust urediniospores survive for a limited period, they rely on the host plant to complete their life cycle. During infection, the host plant induces a defense response to prevent further infection by more virulent physiological races or other pathogenic microorganisms. However, this also alters the host plant's growth and development. When the defense response is induced, energy and nutrient metabolism in the plant changes, reducing nutrient and water uptake and thus inhibiting growth. After stripe rust fungi infect the host plant, symptoms such as yellowing and necrosis appear. When the fungus produces numerous uredinia on the host surface, green leaves and tissues decrease, leading to a significant reduction in photosynthesis. On a macroscopic level, stripe rust infection weakens the host plant's vigor, reducing the number of tillers, spikes, and seeds, and decreasing plant height and seed weight and quality. Economically, stripe rust infection leads to reduced crop yields and increases crop management costs. Agriculturally, stripe rust is a disease of wheat, barley, and some pastures, and can cause epidemics. For wheat, stripe rust epidemics can cause yield losses of 10-70% in normal years, and even total crop failure in severe outbreaks. Planting disease-resistant varieties is considered an effective measure for controlling stripe rust.

[0003] In recent years, with the promotion of superior varieties and improvements in water and fertilizer conditions, wheat production levels have increased significantly. However, due to changes in climate and environment, unreasonable cultivation practices, and variations in the virulence of wheat stripe rust races, the resistance of disease-resistant varieties may be lost, posing a production safety hazard to wheat-growing areas. Using disease-resistant varieties for control remains an effective method. Plant disease-resistant genes can be used to inhibit the infection of plant pathogens; increasing the content of plant disease-resistant proteins will enhance plant disease resistance; and increasing the expression of plant disease-resistant genes will also enhance plant disease resistance.

[0004] Therefore, discovering plant disease resistance genes, understanding the plant response and signal transduction mechanisms under stripe rust stress, and improving plant disease resistance have become important means for the sustainable control of plant stripe rust. Summary of the Invention

[0005] This invention uses genetic engineering to screen for the plant disease resistance-related protein TaCNGC10 and the plant disease resistance-related gene TaCNGC10 that encodes TaCNGC10. By performing functional analysis on the protein and gene, the disease resistance mechanism in the interaction between plants and stripe rust is revealed, providing a solution for the breeding of stripe rust-resistant materials.

[0006] To achieve this technical objective, the present invention adopts the following technical solution.

[0007] In a first aspect, a plant disease resistance-related protein TaCNGC10 is provided, the amino acid sequence of which is as follows:

[0008] 1) As shown in SEQ ID NO:2; or

[0009] 2) A derived amino acid sequence obtained by substituting, deleting and / or adding one or more amino acids from the amino acid sequence in 1), said derived amino acid sequence having the activity of the amino acid sequence shown in 1).

[0010] In this invention, in order to facilitate the purification of the plant disease resistance-related protein TaCNGC10 in 1), a tag as shown in Table 1 can be attached to the amino terminus or carboxyl terminus of the protein composed of the amino acid sequence shown in SEQ ID NO:2.

[0011] In this invention, the plant disease resistance-related protein TaCNGC10 in 2) can be synthesized artificially, or its encoding gene can be synthesized first and then expressed biologically. The encoding gene of the plant disease resistance-related protein TaCNGC10 in 2) can be obtained by deleting one or several amino acid residues from the codons of the plant disease resistance-related gene TaCNGC10 sequence as shown in SEQ ID NO:1, and / or by performing one or several base pairs of missense mutations, and / or by attaching the coding sequence of the tag shown in Table 1 to its 5' end and / or 3' end.

[0012] Table 1. Sequence of Labels

[0013]

[0014] Secondly, a plant disease resistance-related gene TaCNGC10 is provided, wherein the plant disease resistance-related gene TaCNGC10:

[0015] a) Encodes the plant disease resistance-related protein TaCNGC10 described in this invention;

[0016] b) The CDS sequence is shown in SEQ ID NO:1; or

[0017] c) is complementary to a) or b).

[0018] In this invention, the plant disease resistance-related gene TaCNGC10 sequence has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology and encodes a DNA molecule with the same function.

[0019] In this invention, those skilled in the art can readily employ known methods, such as directed evolution and point mutation, to mutate the plant disease resistance-related gene TaCNGC10, which encodes the plant disease resistance-related protein TaCNGC10. Artificially modified polynucleotides that possess 75% or higher sequence identity with the plant disease resistance-related gene TaCNGC10 sequence isolated from this invention, as long as they encode the plant disease resistance-related protein TaCNGC10 and have the same function, are all derived from and equivalent to the plant disease resistance-related gene TaCNGC10 sequence of this invention.

[0020] In this invention, the term "identity" refers to sequence similarity to a natural nucleic acid sequence. "Identity" includes nucleotide sequences having 75% or higher, 85% or higher, 90% or higher, or 95% or higher identity with a polynucleotide sequence encoding a protein as shown in SEQ ID NO:2. Identity can be evaluated visually or using 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. 75% or higher identity can be 80%, 85%, 90%, or 95% or higher.

[0021] Thirdly, a polynucleotide construct or host cell containing the plant disease resistance-related gene TaCNGC10 as described in this invention is provided.

[0022] Fourthly, a recombinant vector, expression cassette, or recombinant bacteria containing the plant disease resistance-related gene TaCNGC10 as described in this invention is provided.

[0023] In this invention, an expression cassette refers to DNA capable of expressing the plant disease resistance-related gene TaCNGC10, as shown in SEQ ID NO:1, in host cells. This DNA may include not only a promoter to initiate polynucleotide transcription as shown in SEQ ID NO:1, but also a terminator to terminate transcription of the plant disease resistance-related gene TaCNGC10, as shown in SEQ ID NO:1. Furthermore, the expression cassette may also include an enhancer sequence. Promoters that can be used in this 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 carmine synthase terminator (NOS terminator), cauliflower mosaic virus CaMV 35S terminator, tml terminator, pea rbcS E9 terminator, and carmine and octopine synthase terminator.

[0024] In this invention, recombinant vectors containing the expression cassette can be constructed using existing expression vectors. The plant expression vectors include vectors suitable for plant microbombardment, such as pAHC25, pBin438, pCAMBIA1302, pCAMBIA2300, pCAMBIA2301, pCAMBIA1305, pCAMBIA1300, pBI121, pCAMBIA1391-Xa, or pCAMBIA1391-Xb (CAMBIA). The plant expression vectors may also contain 3-cell structures of exogenous genes. ’ The terminal untranslated region contains the polyadenylated signal and any other DNA fragments involved in mRNA processing or gene expression. The polyadenylated signal guides the addition of polyadenylated DNA to the 3' end of the mRNA precursor. ’ End-of-cell structures, such as Agrobacterium crown gall induction (Ti) plasmid genes (e.g., the carmine synthase gene Nos), and plant genes (e.g., soybean storage protein genes). ’The untranslated regions of terminal transcription all have similar functions. When constructing plant expression vectors using the genes of this invention, enhancers, including translational enhancers or transcriptional enhancers, can also be used. These enhancer regions can be ATG start codons or adjacent start codons, etc., but they must be identical to the reading frame of the coding sequence to ensure correct translation of the entire sequence. The sources of the translation control signals and start codons are wide-ranging; they can be natural or synthetic. The translation initiation region can originate from the transcription initiation region or structural genes. To facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed. This can involve adding genes that can be expressed in plants, encoding enzymes or luminescent compounds that produce color changes (GUS genes, luciferase genes, etc.), antibiotic marker genes (such as the nptII gene for resistance to kanamycin and related antibiotics, the bar gene for resistance to the herbicide phosphinic acid, the hph gene for resistance to the antibiotic hygromycin, the dhfr gene for resistance to methotrexate, and the EPSPS gene for resistance to glyphosate), or chemical reagent resistance marker genes (such as herbicide resistance genes), and mannose-6-phosphate isomerase genes that provide the ability to metabolize mannose. From a safety perspective, transgenic plants can be screened directly under stress without adding any selective marker genes.

[0025] In this invention, the vector can be a plasmid, a granule, a bacteriophage, or a viral vector.

[0026] In this invention, the microorganism can be yeast, bacteria, algae or fungi, such as Agrobacterium.

[0027] In this invention, the transgenic plant cell lines do not include propagation material.

[0028] Fifthly, the invention provides the application of the polynucleotide constructs or host cells described herein in regulating plant disease resistance.

[0029] Preferably, the disease resistance is resistance to stripe rust.

[0030] Sixthly, the invention provides the application of the polynucleotide constructs or host cells described herein in the cultivation of disease-resistant plants.

[0031] Preferably, the disease resistance is resistance to stripe rust.

[0032] In a seventh aspect, the present invention provides the application of the recombinant vector, expression cassette, or recombinant bacteria described herein in regulating plant disease resistance.

[0033] Preferably, the disease resistance is resistance to stripe rust.

[0034] Eighthly, the invention provides the application of the recombinant vector, expression cassette, or recombinant bacteria described herein in the cultivation of disease-resistant plants.

[0035] Preferably, the disease resistance is resistance to stripe rust.

[0036] Ninthly, a method for cultivating transgenic plants with enhanced disease resistance is provided, the method comprising the following steps:

[0037] 1) Increase the content and / or activity of the plant disease resistance-related protein TaCNGC10 described in this invention in the target plant to obtain transgenic plants;

[0038] 2) Increase the expression of the plant disease resistance-related gene TaCNGC10, which encodes the plant disease resistance-related protein TaCNGC10 described in this invention, in the target plant to obtain transgenic plants;

[0039] The transgenic plant exhibits higher disease resistance than the target plant.

[0040] Preferably, the improvement of the content and / or activity of the plant disease resistance-related protein TaCNGC10 of the present invention in the target plant is achieved by introducing the plant disease resistance-related gene TaCNGC10 of the present invention into the target plant.

[0041] Preferably, the enhancement of the expression of the plant disease resistance-related gene TaCNGC10, which encodes the plant disease resistance-related protein TaCNGC10 of the present invention, in the target plant is achieved by introducing the plant disease resistance-related gene TaCNGC10 of the present invention into the target plant.

[0042] In this invention, the improvement of plant resistance to stripe rust is specifically manifested in any one of the following (1)-(3): (1) Under stripe rust stress, the sporulation of stripe rust infecting transgenic wheat is lower than that of the recipient plant; (2) Under stripe rust stress, the expression level of disease-related genes in transgenic plants is higher than that in recipient plants; (3) Under stripe rust stress, the mycelial infection area of ​​stripe rust infecting transgenic wheat is lower than that in recipient plants. The stripe rust stress is an affinity treatment, and the affinity treatment uses CYR32 and CYR34 strains.

[0043] In this invention, since stripe rust is an obligate parasitic fungus, wheat was chosen as the research object, and the wheat variety (Fielder) was chosen as the recipient of the transgene and the research object.

[0044] In this invention, the gene encoding the plant disease resistance-related protein TaCNGC10 (i.e., the nucleotides shown in SEQ ID NO:1 from 5'1-2085) is introduced into Agrobacterium EHA105 via the recombinant vector CUB-TaCNGC10, which contains an expression cassette of the TaCNGC10 protein encoding gene. The recombinant vector CUB-TaCNGC10 is created by homologous recombination, inserting a DNA fragment of TaCNGC10 into the CUB vector while maintaining other sequences of the CUB vector unchanged. The vector's restriction enzyme site is BamH1.

[0045] In this invention, the transgenic plant is understood to include not only the first-generation transgenic plant obtained by transforming the plant disease resistance-related gene TaCNGC10 into a recipient plant, but also its progeny. For transgenic plants, the gene can be propagated within the species, or it can be transferred into other varieties of the same species using conventional breeding techniques, particularly commercial varieties. The transgenic plant includes seeds, callus tissue, intact plants, and cells.

[0046] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0047] The plant disease resistance-related gene TaCNGC10, verified by this invention, is induced to express by stripe rust fungus. Furthermore, transgenic wheat obtained by introducing TaCNGC10 into wheat exhibits higher resistance to multiple physiological races of stripe rust fungus compared to wild-type wheat. The proteins and genes provided by this invention offer a foundation for the artificial control of disease resistance-related gene expression and will play a crucial role in cultivating broadly resistant plants.

[0048] This invention clarifies the role of the plant disease resistance-related gene TaCNGC10 in the interaction between wheat and stripe rust fungus. Experiments have demonstrated that the plant disease resistance-related gene TaCNGC10 can be used for breeding and improving wheat resistance to stripe rust. The plant disease resistance-related gene TaCNGC10 provided by this invention offers a genetic resource for cultivating wheat materials resistant to stripe rust. Attached Figure Description

[0049] To more clearly illustrate the technical solution of the present invention, the accompanying drawings involved in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the description are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0050] Figure 1 TaCNGC10, a plant disease resistance-related gene ( TaCNGC10 Expression characteristics of wheat in the wheat-striped rust interaction combination.

[0051] Figure 2TaCNGC10, a plant disease resistance-related gene ( TaCNGC10 PCR identification results of wheat T3 generation overexpression.

[0052] Figure 3 TaCNGC10, a plant disease resistance-related gene ( TaCNGC10 Disease resistance identification of genetically modified wheat. Detailed Implementation

[0053] The technical solution of the present invention will be described below with reference to embodiments. However, the present invention is not limited to the following embodiments. Unless otherwise specified, the experimental methods and detection methods described in each embodiment are conventional methods; unless otherwise specified, the reagents and materials can be purchased commercially.

[0054] The wheat stripe rust physiological race CYR23 in this embodiment of the invention has been disclosed in the literature "Liu P, Guo J, Zhang R, et al. TaCIPK10 interacts with and phosphorylates TaNH2 to activate wheat defense responses to stripe rust[J]. Plant biotechnology journal, 2019, 17(5): 956-968." It is publicly available from the applicant.

[0055] The wheat stripe rust race CYR31 in this embodiment of the invention has been disclosed in the literature “Wang Fengle, Wu Liren, Xu Shichang, Jin Shelin, Jia Qiuzhen, Yuan Wenhuan, Yang Jiaxiu. Study on new stripe rust races CYR30 and CYR31 in China [J]. Acta Phytoprotectica Sinica, 1996(01):39-44.” It is available to the public from the applicant.

[0056] The wheat variety Shuiyuan 11 in this embodiment of the invention has been disclosed in the literature “Cao Zhangjun, Jing Jinxue, Wang Meinan, et al. Analysis of the relationship between the stripe rust resistance genes of important domestic resistance varieties Shuiyuan 11, Shuiyuan 92 and Hybrid46 [J]. Northwest Botanical Journal, 2003, 23(1):64-68.” It is available to the public from the applicant.

[0057] Example 1

[0058] This embodiment provides the method for obtaining the plant disease resistance-related protein TaCNGC10 (TaCNGC10) and its encoding gene.

[0059] I. Isolation of mRNA and the plant disease resistance-related gene TaCNGC10 ( TaCNGC10 amplification of )

[0060] Take 7-day-old wheat seedlings of the Shuiyuan 11 variety that are growing normally, quick-freeze them with liquid nitrogen, and store them at -80℃ for later use.

[0061] Total RNA was extracted from wheat leaves using a polysaccharide-polyphenol plant RNA extraction kit (Huayueyang Biotechnology Co., Ltd.). First-strand cDNA synthesis was performed using reverse transcriptase XL (AMV). cDNA was synthesized using the SMART method, and the PCR products were detected by 1.0% agarose gel electrophoresis. The amplification primers were:

[0062] TaCNGC10 -F:5 ’ -ATGTTCGCGCCGAGGAAGGTGGAGG-3 ’ ;

[0063] TaCNGC10 -R:5 ’ -CTCGTAGGGTTCGACCGAGAAATCT-3 ’ .

[0064] A 2085 bp PCR product was obtained. Sequencing revealed that this PCR product contained the nucleotides shown in positions 1-2085 of SEQ ID NO:1, and the gene represented by these nucleotides was named the plant disease resistance-related gene TaCNGC10. TaCNGC10 ); TaCNGC10 The amino acid sequence of the encoded protein is shown in SEQ ID NO:2, and the protein is named plant disease resistance-related protein TaCNGC10 (TaCNGC10).

[0065] II. RT-PCR Detection TaCNGC10 Expression induced by stripe rust fungi

[0066] 1. Preparation of experimental materials

[0067] Stripe rust inoculation was performed according to the method described by Kang Zhensheng et al. (1984, Journal of Northwest Agricultural College). Wheat variety Shuiyuan 11 leaves were inoculated with CYR23 (incompatible) or CYR31 (compatible) to form incompatible and compatible interaction combinations, respectively, with sterile water used as a control.

[0068] Samples were taken at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h, 96 h, and 120 h post-inoculation, with the control sampling time points consistent with the treatment. Fresh leaves were cut, wrapped in aluminum foil, flash-frozen in liquid nitrogen, and then stored at -80℃ for later use. Total RNA was extracted from wheat leaves using the Trizol method, and first-strand cDNA was synthesized using reverse transcriptase XL (AMV). cDNA was synthesized using the SMART method.

[0069] 2. RT-qPCR detection TaCNGC10expression level

[0070] According to wheat TaCNGC10 and elongation factor gene TaEF We designed specific quantitative PCR primers based on the sequence (GenBank accession number: U76744).

[0071] The RT-qPCR primer sequences are:

[0072] TaCNGC10 -RT-F:5 ’ -GGTGAAGATGACCCTCCACCCAAG-3 ’ ;

[0073] TaCNGC10 -RT-R:5'-CTCGTAGGGTTCGACCGAGAAATCT-3 ’ ;

[0074] TaEF -F:5 ’ -TGGTGTCATCAAGCCTGGTATGGT-3 ’ ;

[0075] TaEF -R:5 ’ -ACTCATGGTGCATCTCAACGGACT-3 ’ .

[0076] Before use, the specificity and amplification efficiency (≥90%) of the amplification products of the quantitative PCR primers must be tested. TaEF-1α was used as an internal reference gene in real-time PCR analysis. Using an AceQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) and a Bio-Rad CFX Manager quantitative PCR instrument (Bio-rad, Hercules, California), and following the instructions, real-time quantitative PCR amplification was performed using cDNA from each treatment sampling site as a template. Each reaction was performed in at least three replicates. The Ct values, mean, and standard deviation of each replicate were generated by the quantitative PCR instrument by manually adjusting the baseline. Each reaction was performed in three replicates, and the average Ct value was used. The Delta-Ct method was used to analyze the experimental data to determine the relative expression level of the gene.

[0077] The results of RT-qPCR are as follows Figure 1 As shown, TaCNGC10 The expression patterns of wheat variety Shuiyuan 11 after inoculation with the non-compatible race CYR23 and the compatible race CYR31 of stripe rust, respectively, were analyzed. TaCNGC10Upregulated expression was observed in both the incompatible and compatible combinations during the pre-infection phase after inoculation.

[0078] The above results indicate that TaCNGC10 Expression induced by stripe rust fungi.

[0079] Example 2

[0080] This embodiment provides TaCNGC10 Application in improving plant resistance to stripe rust

[0081] one, TaCNGC10 Obtaining genetically modified wheat

[0082] 1. TaCNGC10 Construction of overexpression vectors

[0083] The amplified in Example 1 TaCNGC10 From 5 ’ The 1-2085 nucleotide fragment was ligated into the CUB vector digested with BamHI using the ClonExpress II OneStep Cloning Kit (vazyme) for homologous recombination, resulting in the recombinant vector. TaCNGC10 -CUB.

[0084] TaCNGC10 The specific construction method of the CUB carrier is as follows:

[0085] Amplified TaCNGC10 Using a full-length CDS as a template, the following primers were used for amplification, and the resulting fragment was directly recombined into the CUB vector.

[0086] TaCNGC10 -CUB-F:

[0087] 5 ’ -CAGGTCGACTCTAGA GGATCC ATGTTCGCGCCGAGGAAGGTGGAGG-3 ’ ;

[0088] TaCNGC10 -CUB-R:

[0089] 5 ’ -GAGCTCGGTACCCGG GGATCC CTCGTAGGGTTCGACCGAGAAATCT-3 ’ ;

[0090] For TaCNGC10 Transgenic wheat transformation uses BamHI as the restriction enzyme site. The underlined part in the primers above indicates the restriction enzyme site.

[0091] 2. TaCNGC10Preparation of genetically modified wheat

[0092] Recombinant vector TaCNGC10 -CUB used Agrobacterium to infect wild-type wheat Fielder callus to obtain T0 generation. TaCNGC10 Genetically modified wheat.

[0093] Cultivating T0 generation TaCNGC10 Genetically modified wheat until the T3 generation was obtained TaCNGC10 Two strains of genetically modified wheat.

[0094] 3. PCR verification

[0095] T3 generation was extracted using the CTAB method. TaCNGC10 Genomic DNA from leaves of two genetically modified wheat lines and the wild-type control wheat (Fielder) was used. TaCNGC10 Overexpression detection F: 5 ’ -CATCGTCAACTTCCTGATGAATTGC-3 ’ BESNOS2-R: 5 ’ -TTGCGGGACTCTAATCATAAAAAC-3 ’ Molecular detection was performed on transgenic T3 generation plants (upstream primer located on the gene, downstream primer located on the NOS terminator). Five plants were randomly selected from each line. Water was used as a blank control, and DL2000 Plus DNA Marker (vazyme) was used as a standard. The standard indicated the size as shown in the figure. Figure 2 As shown.

[0096] The results are as follows Figure 2 As shown, the line yielding 846bp is the positive strain, indicating overexpression. TaCNGC10 The two strains of the T3 generation ( TaCNGC10 Overexpression lines 13 and TaCNGC10 All 46 overexpression lines were positive and were named... TaCNGC10 Overexpression line 13 TaCNGC10 Overexpression line 46.

[0097] II. Transfer TaCNGC10 Analysis of stripe rust resistance in genetically modified wheat

[0098] Wild-type wheat Fielder (material sourced from the College of Plant Protection, Northwest A&F University) TaCNGC10 Overexpression lines 13 and TaCNGC10 Overexpression line 46 was cultured in an incubator with a day / night temperature of 25 / 23℃ and a light / dark cycle of 16 h light / 8 h dark. After the second leaf unfolded... Figure 3In section A), the leaves were inoculated with the physiological races CYR32 and CYR34 of stripe rust (the strains were obtained from the College of Plant Protection, Northwest A&F University). The inoculation method was in accordance with the literature "Kang Zhensheng, Li Zhenqi. Discovery of a new pathogenic strain of Lovlin 10 at room temperature [J]. Journal of Northwest A&F University (Natural Science Edition) 1984(04):18-28." 120 h after inoculation, the leaves at the inoculation site were taken as RNA extraction samples, and the disease phenotype was observed 14 days after inoculation.

[0099] Phenotypic results are as follows Figure 3 As shown in Figure A, under the infection conditions of stripe rust fungi CYR32 and CYR34, a large number of spore masses were observed on the leaves of all treatments in the control plants, while TaCNGC10 Overexpression line 13 TaCNGC10 The overexpression line 46 showed a significant decrease in the number of uredinia in wheat leaves and an increase in the area of ​​chlorotic necrosis. Fielder and other pathogens were also detected during stripe rust infection. TaCNGC10 Overexpression lines 13 and TaCNGC10 46 overexpression lines TaCNGC10 The expression level was detected using RNA from different inoculation sites 120 h post-inoculation as templates, following the RT-PCR detection method described in Example 1. The results are as follows: Figure 3 As shown in Figure B, in the overexpression material TaCNGC10 The expression level was consistently more than 5 times higher than that of the control.

[0100] III. Biomass Detection

[0101] Stripe rust biomass was measured in wheat 7 days after inoculation. The biomass measurement method followed the guidelines in the literature "Qi Tuo et al., Stripe Rust Effector PstGSRE1 Disrupts Nuclear Localization of ROS-Promoting Transcription Factor TaLOL2 to Defeat ROS-Induced Defense in Wheat, 2019". Results are as follows: Figure 3 As shown in Figure C, the biomass of stripe rust fungus in wheat was significantly reduced after overexpression.

[0102] The above disease resistance identification results show that, under the treatment conditions of CYR32 and CYR34 physiological races, transgenic wheat exhibits strong resistance. This has significant advantages in production applications.

[0103] This proves TaCNGC10 It is an important gene involved in the process of wheat resistance to stripe rust.

[0104] The embodiments described above are some, but not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but rather to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art through related deductions and substitutions based on the inventive concept, without inventive effort, are within the scope of protection of the present invention.

Claims

1. The application of recombinant vectors, expression cassettes, or recombinant bacteria in regulating plant resistance to stripe rust, wherein the plant is wheat; The recombinant vector, expression cassette, or recombinant bacteria contains the plant disease resistance-related gene TaCNGC10; The plant disease resistance-related gene TaCNGC10: a) encodes the plant disease resistance-related protein TaCNGC10; b) its CDS sequence is shown in SEQ ID NO:1; The amino acid sequence of the plant disease resistance-related protein TaCNGC10 is shown in SEQ ID NO:2; The application is to increase the content and / or activity of TaCNGC10, a plant disease resistance-related protein, in plants.

2. The application of recombinant vectors, expression cassettes, or recombinant bacteria in the cultivation of stripe rust-resistant plants, wherein the plant is wheat; The recombinant vector, expression cassette, or recombinant bacteria contains the plant disease resistance-related gene TaCNGC10; The plant disease resistance-related gene TaCNGC10: a) encodes the plant disease resistance-related protein TaCNGC10; b) its CDS sequence is shown in SEQ ID NO:1; The amino acid sequence of the plant disease resistance-related protein TaCNGC10 is shown in SEQ ID NO:2; The application is to increase the content and / or activity of TaCNGC10, a plant disease resistance-related protein, in plants.

3. A method for cultivating transgenic plants with enhanced disease resistance, characterized in that, The method includes the following steps: 1) Increase the content and / or activity of the plant disease resistance-related protein TaCNGC10 in the target plant to obtain transgenic plants; 2) Increase the expression of the plant disease resistance-related gene TaCNGC10, which encodes the plant disease resistance-related protein TaCNGC10, in the target plant to obtain transgenic plants; The transgenic plant exhibits higher disease resistance than the target plant. The disease resistance is resistance to stripe rust; the plant is wheat; The amino acid sequence of the plant disease resistance-related protein TaCNGC10 is shown in SEQ ID NO:2; The plant disease resistance-related gene TaCNGC10: a) encodes the plant disease resistance-related protein TaCNGC10; b) its CDS sequence is shown in SEQ ID NO:1.

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