Application of GhGDSL1 protein and / or GhTLP protein in improving plant resistance

Abstract

The invention provides the application of the GhGDSL1 protein and / or the GhTLP protein in improving the plant resistance, the synergistic effect and the application value of the GhGDSL1 protein and the GhTLP protein in plant verticillium dahliae resistance reaction are determined for the first time, the resistance of target plants such as cotton and the like can be efficiently improved by means of multiple implementation modes such as protein direct application, gene introduction or overexpression and the like, and the application of the GhGDSL1 protein and / or the GhTLP protein in improving the plant resistance is realized. The method is of great significance to safe production of target plants and promotion of green and sustainable development of agriculture.

Description

Technical Field

[0001] This application relates to the field of biotechnology, specifically to the application of GhGDSL1 protein and / or GhTLP protein in enhancing plant resistance, a method for enhancing plant resistance, and the application of GhGDSL1 protein and / or GhTLP protein in the preparation of pesticides. Background Technology

[0002] Cotton is one of the world's most important natural fiber and oilseed crops, and its safe production is crucial to the national economy. However, it is susceptible to damage caused by Verticillium dahliae (…). Verticillium dahliae Verticillium wilt, caused by cotton, is a devastating soil-borne vascular disease, often referred to as the "cancer" of cotton. Typical symptoms of Verticillium wilt in cotton include yellowing, wilting, drying, and shedding of leaves, as well as stunted boll development and even boll shedding, leading to a sharp decline in yield and quality, severely restricting the sustainable development of the cotton industry.

[0003] In the field of cotton breeding, "Zhongzhimian No. 2" is a breakthrough variety, possessing excellent characteristics of resistance to Verticillium wilt, immunity to Fusarium wilt, and high resistance to bollworm. It has been listed as a key variety by the Ministry of Agriculture and Rural Affairs and has long served as a control variety for cotton Verticillium wilt resistance breeding in regional trials. However, despite its clearly defined disease-resistant phenotype, its intrinsic molecular resistance mechanism, particularly the key genes responsible for its outstanding Verticillium wilt resistance, has not been fully elucidated. This limits the ability to further explore and utilize its disease-resistant gene resources based on this superior germplasm.

[0004] In current technologies, researchers commonly employ genomics, transcriptomics, and bioinformatics to discover disease resistance genes in plants. For example, differential analysis of transcriptome data from resistant and susceptible cotton varieties (such as Zhongzhimian 2 and Junmian 1) after infection with Verticillium dahliae, combined with secretory proteome prediction, is an effective strategy for screening candidate disease resistance genes. Secretory proteins, especially those located on apoplasts (the first line of defense against pathogen invasion), play a crucial role in plant immunity by recognizing pathogens, transmitting signals, and directly resisting fungi.

[0005] While existing technologies have identified several Verticillium wilt-related genes in cotton (such as GhRVW2 and GhEB1C) and recognized the roles of secreted proteins GDSL esterases and TLP family proteins in disease resistance, significant shortcomings remain. First, the core genetic basis of disease resistance in the key resistant variety "Zhongzhimian 2" is still incomplete. Second, current research largely focuses on the function of individual genes, while the understanding of how multiple key genes interact synergistically to form a complex resistance network is still lacking. Specifically, no publicly available technology has yet revealed a direct protein-protein interaction between GDSL esterase / lipase family genes and sweet protein (TLP) in the Verticillium dahliae resistance pathway, and their joint regulation of cotton immune responses.

[0006] Therefore, there is an urgent need in this field to identify new, key disease-resistant genes that play a core role in Zhongzhimian 2 and to further reveal the molecular mechanisms by which they exert their disease-resistant functions, especially their interaction networks with other proteins. This is of great significance for elucidating the source of Zhongzhimian 2's superior resistance, enriching the theory of plant disease resistance, and developing new strategies for the control of Verticillium wilt. Summary of the Invention

[0007] The purpose of this disclosure is to provide the application of GhGDSL1 protein and / or GhTLP protein in improving plant resistance to Verticillium dahliae, to reveal the synergistic interaction of the two in disease resistance molecular mechanisms, and to provide key gene resources and theoretical support for molecular breeding of cotton and other plants to resist Verticillium wilt and the development of new biological pesticides.

[0008] To achieve the above objectives, the first aspect of this disclosure provides the application of GhGDSL1 protein and / or GhTLP protein in enhancing plant resistance, wherein the amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; The amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2.

[0009] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae.

[0010] Optionally, the plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0011] On the other hand, this disclosure provides a method for improving plant resistance, the method comprising the following steps: Apply the GhGDSL1 protein and / or GhTLP protein to the target plant, and / or introduce the gene encoding the GhGDSL1 protein and / or GhTLP protein into the target plant, and / or cause the GhGDSL1 protein and / or GhTLP protein to be overexpressed in the target plant. The amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2; The nucleotide sequence of the gene encoding the GhGDSL1 protein is shown in SEQ ID NO.3; The nucleotide sequence of the gene encoding the GhTLP protein is shown in SEQ ID NO.4.

[0012] Optionally, the genes encoding GhGDSL1 protein and / or GhTLP protein are introduced into the target plant via a plant expression vector.

[0013] Optionally, the plant expression vector includes pCAMBIA1300-35S-GhGDSL1 and pCAMBIA1300-35S-GhTLP.

[0014] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae.

[0015] Optionally, the plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0016] On the other hand, this disclosure provides the use of GhGDSL1 protein and / or GhTLP protein in the preparation of pesticides for improving plant resistance, wherein the amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; and the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2.

[0017] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae; The plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0018] Through the above technical solutions, this disclosure clarifies for the first time the synergistic effect and application value of GhGDSL1 protein and GhTLP protein in plant resistance to Verticillium dahliae. By using multiple methods such as direct protein application, gene introduction or overexpression, the resistance of target plants such as cotton can be effectively improved, which is of great significance for the safe production of target plants and the promotion of green and sustainable agricultural development.

[0019] Other features and advantages of this disclosure will be described in detail in the following detailed description section. Attached Figure Description

[0020] The accompanying drawings are provided to further illustrate the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof. In the drawings: Figure 1This is a diagram showing the cloning and sequence structure analysis of the GhGDSL1 gene; Figure 2 This is a subcellular localization map of the secretory activity of the GhGDSL1 signal peptide; Figure 3 This is a subcellular localization map of the GhGDSL1 protein; Figure 4 This is a graph showing the effect of VIGS silencing GhGDSL1 on cotton resistance to Verticillium wilt; Figure 5 This is a graph showing the effect of GhGDSL1 overexpression in Arabidopsis thaliana on resistance to Verticillium dahliae; Figure 6 This is an immunoprecipitation verification diagram of GhGDSL1 and GhTLP proteins. Detailed Implementation

[0021] The following provides a detailed description of specific embodiments of this disclosure. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit this disclosure.

[0022] On the one hand, this disclosure provides the application of GhGDSL1 protein and / or GhTLP protein in improving plant resistance, wherein the amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; The amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2.

[0023] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae.

[0024] Optionally, the plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0025] On the other hand, this disclosure provides a method for improving plant resistance, the method comprising the following steps: Apply the GhGDSL1 protein and / or GhTLP protein to the target plant, and / or introduce the gene encoding the GhGDSL1 protein and / or GhTLP protein into the target plant, and / or cause the GhGDSL1 protein and / or GhTLP protein to be overexpressed in the target plant. The amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2; The nucleotide sequence of the gene encoding the GhGDSL1 protein is shown in SEQ ID NO.3; The nucleotide sequence of the gene encoding the GhTLP protein is shown in SEQ ID NO.4.

[0026] Optionally, the genes encoding GhGDSL1 protein and / or GhTLP protein are introduced into the target plant via a plant expression vector.

[0027] Optionally, the plant expression vector includes pCAMBIA1300-35S-GhGDSL1 and pCAMBIA1300-35S-GhTLP.

[0028] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae.

[0029] Optionally, the plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0030] On the other hand, this disclosure provides the use of GhGDSL1 protein and / or GhTLP protein in the preparation of pesticides for improving plant resistance, wherein the amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; and the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2.

[0031] Optionally, the enhancement of plant resistance includes: enhancing the plant's resistance to Verticillium dahliae; The plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

[0032] The present disclosure is further described in detail below through examples. All raw materials used in the examples are commercially available.

[0033] SEQ ID NO.1 (Amino acid sequence of GhGDSL1 protein): MAKTCFHGVFVSIAVTSLLLLQAGAPPPPPPAVYIFGDSTLDVGTNNFIPECAARADFYFNGIDFPYSKPTGRFSNGLNTADQIVRLLGLKKSPPPFLYLVNDPSNFQKNILQGANFASGGSGVLRDTGKAKRVIPLEEQIQHFSTIRSNITNMTGSEEATDKILSKAFILISIGSNDMFEY LLNLSKPMSLAEFNATLISTYEYHIKTLYELGARTFGILTVPPIGCTPFARAVFTGNNSCFEPAQAMAVQFYFDVGSSLEQFSSTVQDMKYSVGNTFLMTGVLTGDMLAFGLKNIAAACCGNGTYGCNQTASFCSNRDEYLFWDQFHPTQRASELVALTLFGAAEPIMVPMNFSQLLGVNI; SEQ ID NO.2 (Amino acid sequence of GhTLP protein): MKLESLSPFFFPFVALCFTVATAATFTIRNNCSYTVWAAAVAASGGGGGKRLESGATWNLNVKPGTRGARIWARTNCQFDEAGQGRCKTGDCGGRLKCKAYGNPPNTLAEFALNQYKNLDFF HISLVDGFNVPMEFSPTSGLCKKGIQCTADIVRQCPKELEAPGGCNDPCTVFKTQKYCCYYGKCGPTDFSKFFKARCPSAYTYPRDDPSSTVTCPGGTNYKVVFCPIGSPHLEMVASMSQEE; SEQ ID NO.3 (nucleotide sequence encoding GhGDSL1 protein): SEQ ID NO.4 (nucleotide sequence encoding GhTLP protein): ATGAAGTTGGAAAGCCTCTCTCCGTTTTTCTTCCCTTTCGTTGCTCTTTGCTTCACGGTGGCCACTGCTGCCACTTTTACCATTCGAAACAACTGCTCCTACACGGTCTGGGCAGCAGCCGTGGCCGCCTCCGGTGGTGGTGGTGGTAAGAGGCTAGAGAGCGGTGCAACTTGGAATCTAAAT GTGAAGCCCGGCACTAGAGGAGCACGTATCTGGGCTCGAACCAATTGCCAATTCGATGAAGCCGGACAAGGCAGATGCAAAACCGGTGATTGTGGGGGGCGACTTAAGTGCAAAGCCTATGGTAATCCCCCAAACACCTTGGCTGAATTCGCACTAAACCAGTACAAAAACTTGGATTTCTTCC ATATCTCTCTGGTTGATGGATTTAATGTTCCTATGGAGTTTAGCCCAACCTCTGGGTTGTGCAAGAAGGGCATTCAATGCACAGCTGATATAGTAAGGCAGTGCCCTAAGGAATTGGAAGCCCCAGGTGGGTGCAATGATCCTTGCACCGTTTTCAAGACTCAAAAATATTGTTGCTATTACGG GAAGTGTGGCCCAACGGATTTTTCCAAGTTCTTCAAGGCAAGATGCCCCAGTGCTTATACTTATCCACGAGATGATCCTTCAAGCACAGTCACTTGCCCTGGTGGAACCAATTACAAGGTTGTGTTCTGCCCCATAGGCTCTCCTCATCTAGAGATGGTTGCAAGCATGAGCCAAGAAGAGTAA Example 1 This embodiment is used to clone the full-length CDS of the cotton GhGDSL1 gene and analyze its structural features.

[0034] RNA extraction and cDNA synthesis: Root tissue of Zhongzhi Mian No. 2 seedlings was taken, and total RNA was extracted using the Trizol method. cDNA was then synthesized by reverse transcription (PrimeScript RT reagent Kit, TaKaRa).

[0035] Gene cloning: Specific primers (upstream primer: ATGGCAAAGACATGTTTTCATGSEQ ID NO.5, downstream primer: CTAAATGTTAACCCCCAGCAACSEQ ID NO.6) were designed based on the transgenic data of Zhongzhimian No. 2, and PCR amplification was performed using cDNA as a template. The amplified products were verified by agarose gel electrophoresis (results are shown in Figure 1). Figure 1 As shown in Figure A, where "M" represents the DNA molecular weight standard, and the band in the cDNA lane corresponds to the cDNA amplification product of the GhGDSL1 gene (the size is consistent with expectations), it is ligated into the TA-Zero vector, transformed into E. coli DH5α, and positive clones are selected for sequencing.

[0036] Sequence analysis: Sequencing results showed that the full-length CDS of GhGDSL1 is 1458 bp (nucleotide sequence as shown in SEQ ID NO. 3), encoding a protein containing 363 amino acids (amino acid sequence as shown in SEQ ID NO. 1). Gene structure analysis revealed that its open reading frame consists of four exons with lengths of 253 bp, 140 bp, 231 bp, and 468 bp (as shown in SEQ ID NO. 1). Figure 1 (As shown in B); Protein domain prediction indicates that the protein contains a signal peptide (SP, 1-24aa) at its N-terminus, and a conserved GDSL esterase domain (Lipase_GDSL, containing the GDSL motif and key cysteine ​​residues such as Cys242 and Cys302) in its core region. Figure 1 As shown in C), it belongs to the typical GDSL esterase / lipase family.

[0037] Example 2 Verification of the secretory activity of GhGDSL1 signal peptide This embodiment uses a recombinant vector to construct and verify the secretion function of the GhGDSL1 signal peptide.

[0038] (1) Vector construction: The GhGDSL1 signal peptide sequence was inserted into the pSUC2 secretion function verification vector to construct pSUC2-GhGDSL1. SP Recombinant vectors; simultaneously, YTK12 blank strain, pSUC2 empty vector, and Avr1b were set up. SP Positive control.

[0039] (2) Functional assay: Each vector was transformed into a yeast strain, and after culturing, TTC (triphenyltetrazolium chloride) was added for color development. The color change of the solution was observed, and the results were as follows: Figure 2 As shown: YTK12 and pSUC2 control tube solutions are clear; Avr1b SP The positive control tube was reddish-brown; GhGDSL1 SPThe recombinant vector tube was bright red, consistent with the trend of the positive control, further confirming that its signal peptide has secretory activity, which can drive protein secretion and trigger the TTC reduction reaction.

[0040] The above experiments demonstrate that the GhGDSL1 signal peptide has the function of guiding the secretion of proteins into the extracellular space, providing a structural basis for its disease-fighting effect in the apoplast.

[0041] Example 3 This embodiment clarifies the localization of the GhGDSL1 protein in plant cells.

[0042] Vector construction: The full-length CDS of GhGDSL1 was fused with the GFP gene to construct the pCAMBIA1300-GhGDSL1-GFP recombinant vector, with the empty vector GFP as a control.

[0043] Transient transformation in tobacco: Recombinant and control vectors were injected into tobacco leaves using Agrobacterium-mediated genetic transformation. Fluorescence was observed after 48 hours of incubation. Results are as follows: Figure 3 As shown, the fluorescence signal of the GhGDSL1 group (pCAMBIA1300-GhGDSL1-GFP) is mainly concentrated in the extracellular apoplast region, consistent with apoplast-specific labeling characteristics (such as the fluorescence distribution pattern on the outer side of the cell membrane); the fluorescence signal of the empty vector GFP group is widely distributed throughout the cell. This comparison confirms that the GhGDSL1 protein is specifically localized in the apoplast.

[0044] Example 4 This embodiment verifies the disease resistance function of GhGDSL1 by virus-induced gene silencing (VIGS).

[0045] Construction of silencing vector: A specific fragment (350 bp) of GhGDSL1 was amplified and inserted into the pTRV2 vector to construct the TRV2-GhGDSL1 silencing vector; the empty TRV2 vector was used as a control.

[0046] Cotton infection treatment: Agrobacterium GV3101 was co-transformed with the recombinant vector and pTRV1 helper vector and injected into the cotyledons of cotton seedlings No. 2. After 2 weeks of culture, the silencing efficiency was detected by qPCR. The results showed that the expression level of GhGDSL1 was reduced by 82%, and the silencing effect was significant.

[0047] Verticillium wilt resistance identification: Silent plants and control plants were inoculated with Verticillium dahliae (concentration 1×10⁻⁶). 6 (spores / mL), and the incidence of disease was investigated 21 days later.

[0048] Results: Phenotypic observation: Plants in the TRV2:GhGDSL1 group showed obvious leaf yellowing, wilting, and even leaf drop. Figure 4A and D) showed significantly higher browning of the vascular bundles in the stems compared to the control. Figure 4 A); while the WT and TRV2:00 groups showed normal phenotypes.

[0049] Disease index statistics: Disease index of TRV2:GhGDSL1 group ( Figure 4 E) The disease index was significantly elevated, with most plants reaching a disease level of 3-4; the disease index of the WT and TRV2:00 groups was below level 1.

[0050] Pathogen colonization detection: Fungal DNA was extracted from stem base tissue, and the biomass of *Verticillium dahliae* was detected by qPCR. Figure 4 The results showed that the relative biomass of pathogens in the TRV2:GhGDSL1 group was more than 8 times that of the TRV2:00 group (P<0.001), and in plate culture ( Figure 4 F) Numerous pathogen colonies were observed, further confirming that silencing GhGDSL1 significantly reduced cotton's resistance to Verticillium dahliae.

[0051] Example 5 This embodiment verifies the disease resistance function of GhGDSL1 by overexpression in Arabidopsis thaliana.

[0052] Overexpression vector construction and plant selection: The full-length CDS of GhGDSL1 (nucleotide sequence shown in SEQ ID NO.3) was inserted into the pBI121 vector (driven by the CaMV 35S promoter), and Arabidopsis thaliana (Col-0 ecotype) was transformed using Agrobacterium-mediated transformation. Homozygous overexpression lines OE1, OE2, and OE3 were obtained after hygromycin (HYG) resistance screening. Figure 5 A); with Col-0 wild-type (WT) and mutant gds1 For comparison.

[0053] Overexpression efficiency verification: RNA was extracted from leaves of each strain and analyzed by PCR ( Figure 5 B) and qPCR Figure 5 C) The expression level of GhGDSL1 was detected. The results showed that the relative expression level of GhGDSL1 in the OE1, OE2, and OE3 lines was significantly higher than that in WT (P<0.001), with the OE2 line showing the highest expression level, confirming that the overexpression vector was successfully constructed and its expression was stable.

[0054] Results: For overexpression lines, WT and... gds1 The patients were inoculated with Verticillium dahliae (strain Vd991) and their phenotypes were observed and the incidence of disease was recorded 14 days later.

[0055] Phenotypic observation: Under the Mock treatment (uninoculated), all lines grew normally; after inoculation with Vd991, WT and gds1 The leaves of the strain showed obvious wilting and yellowing. Figure 5 D), while the disease severity of the overexpression lines OE1, OE2, and OE3 was significantly lower than that of WT, with the OE2 line showing the strongest resistance.

[0056] Pathogen colonization detection: Fungal DNA was extracted from root tissue, and the biomass of *Verticillium dahliae* was detected by qPCR. Figure 5 E). The results showed that the relative biomass of the pathogen in the overexpression lines was significantly lower than that in the WT (P<0.01), and gds1 The biomass of the genotype was significantly higher than that of the control group (P<0.01), further confirming that GhGDSL1 overexpression can significantly improve the resistance of Arabidopsis thaliana to Verticillium dahliae.

[0057] In summary, overexpression of GhGDSL1 in Arabidopsis thaliana effectively enhances its resistance to Verticillium dahliae, indicating that this gene has a conserved function in the plant resistance pathway to Verticillium wilt.

[0058] Example 6 Verification of the interaction between GhGDSL1 and GhTLP proteins This embodiment verifies the interaction between GhGDSL1 and GhTLP through an immunoprecipitation experiment.

[0059] Vector construction and transient expression in tobacco: The GhTLP gene (amino acid sequence shown in SEQ ID NO.2, nucleotide sequence of its encoding gene shown in SEQ ID NO.4) was fused with an HA tag to construct the GhTLP-HA vector; the GhGDSL1 gene was fused with a GFP tag to construct the GhGDSL1-GFP vector; and an empty GFP vector was constructed as a control. Using Agrobacterium-mediated transformation, GhTLP-HA was co-transformed with either GhGDSL1-GFP or GFP, and cultured for 48 hours.

[0060] Protein extraction and immunoprecipitation: Tobacco leaf tissue was collected and total protein was extracted; GhTLP-HA was immunoprecipitated using HA antibody-conjugated magnetic beads (Input is the total protein sample loaded, Co-IP is the sample after precipitation).

[0061] Western Blot analysis: The expression of GHTLP-HA was detected using an anti-HA antibody (specific bands were visible in both the input and Co-IP lanes); interacting proteins were detected using an anti-GFP antibody, and the results are as follows: Figure 6 As shown: The specific band of GhGDSL1-GFP (approximately 75 kDa) can be detected only in Co-IP samples co-transformed with GhTLP-HA and GhGDSL1-GFP, while this band is not present in samples co-transformed with GhTLP-HA and empty vector GFP.

[0062] In summary, the immunoprecipitation experiment confirmed the direct protein-protein interaction between GhGDSL1 and GhTLP in plants, providing key evidence for the molecular mechanism by which the two synergistically regulate cotton resistance to Verticillium dahliae.

[0063] The preferred embodiments of this disclosure have been described in detail above. However, this disclosure is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this disclosure, various simple modifications can be made to the technical solutions of this disclosure, and these simple modifications all fall within the protection scope of this disclosure.

[0064] It should also be noted that the various specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, this disclosure will not describe the various possible combinations separately.

[0065] Furthermore, various different embodiments of this disclosure can be combined in any way, as long as they do not violate the spirit of this disclosure, they should also be regarded as the content disclosed in this disclosure.

Claims

1. The application of GhGDSL1 protein and / or GhTLP protein in enhancing plant resistance, characterized in that, The amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; The amino acid sequence of the GhTLP protein is shown in SEQ ID NO.

2.

2. The application according to claim 1, wherein, The improvement of plant resistance includes: improving the plant's resistance to Verticillium dahliae.

3. The application according to claim 2, wherein, The plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

4. A method for improving plant resistance, characterized in that, The method includes the following steps: Apply the GhGDSL1 protein and / or GhTLP protein to the target plant, and / or introduce the gene encoding the GhGDSL1 protein and / or GhTLP protein into the target plant, and / or cause the GhGDSL1 protein and / or GhTLP protein to be overexpressed in the target plant. The amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.2; The nucleotide sequence of the gene encoding the GhGDSL1 protein is shown in SEQ ID NO.3; The nucleotide sequence of the gene encoding the GhTLP protein is shown in SEQ ID NO.

4.

5. The method according to claim 4, wherein, The genes encoding GhGDSL1 and / or GhTLP proteins are introduced into the target plant via a plant expression vector.

6. The method according to claim 5, wherein, The plant expression vectors include pCAMBIA1300-35S-GhGDSL1 and pCAMBIA1300-35S-GhTLP.

7. The method according to claim 4, wherein, The improvement of plant resistance includes: improving the plant's resistance to Verticillium dahliae.

8. The method according to claim 7, wherein, The plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

9. The application of GhGDSL1 protein and / or GhTLP protein in the preparation of pesticides, characterized in that, The pesticide is used to improve plant resistance. The amino acid sequence of the GhGDSL1 protein is shown in SEQ ID NO.1; the amino acid sequence of the GhTLP protein is shown in SEQ ID NO.

2.

10. The application according to claim 9, wherein, The improvement of plant resistance includes: improving the plant's resistance to Verticillium dahliae; The plant includes at least one of cotton, Arabidopsis thaliana, and tomato.

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