Method for synergistically regulating cotton fiber elongation and verticillium wilt resistance based on GhMP gene of upland cotton and transgenic cotton plant

Abstract

The invention provides a method for synergistically regulating cotton fiber elongation and verticillium wilt resistance based on an upland cotton GhMP gene and a transgenic cotton plant, and relates to the technical field of genetic engineering. The gene encodes a membrane protein, and overexpression and RNA interference transgenic line experiments prove that the expression level of GhMP is positively correlated with fiber development and disease resistance: the fiber length can be significantly increased and the verticillium wilt resistance can be improved by expression increase, and the fiber shortening and the resistance weakening can be caused by expression reduction. A further research shows that the GhMP protein is positioned on a cell membrane, can interact with GhPIP2.7 and GhTIP1.1 aquaporin, and jointly participates in hydrogen peroxide transmembrane transport to form a molecular compound for regulating and controlling fiber elongation and preventing diseases. The invention provides a key target gene with both quality improvement and disease resistance enhancement, provides a new strategy and gene resources for cotton multi-trait molecular breeding, and has important application value and popularization prospect.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, and in particular to a method based on upland cotton. GhMP Methods for gene-coordinated regulation of cotton fiber elongation and Verticillium wilt resistance, and transgenic cotton plants. Background Technology

[0002] Cotton is the world's most important natural fiber crop, with my country accounting for over 20% of global cotton production. Fiber length and resistance to Verticillium wilt are core traits determining cotton yield and quality. Cotton fiber quality indicators include more than ten factors such as length, strength, fineness, maturity, and uniformity, among which fiber length is one of the key traits determining cotton quality and economic benefits. For every 1 mm increase in fiber length, yarn strength can increase by 2%-3%, and long-fiber cotton (≥28 mm) is more efficient in spinning than short-fiber cotton (<25 mm), thus reducing costs. In recent years, with rising living standards, the demand for high-end textiles has increased significantly. However, some high-quality cotton varieties in China still lag behind international advanced levels, making it difficult to fully meet the demands of the high-end textile market. High-quality raw cotton suitable for spinning yarns of 80 count and above still relies to some extent on imports. Therefore, utilizing modern biotechnology through molecular breeding to improve cotton fiber quality, especially fiber length, is not only an urgent industry need but also a national policy direction.

[0003] Verticillium wilt of cotton ( Verticillium dahliae Verticillium wilt (Variegata wilt) is a semi-living, soil-borne fungal pathogen that can infect a variety of plants. It infects plants through the roots, multiplies in the vascular system, and causes wilting symptoms in the host. Variegata wilt causes significant yield losses in cotton-growing areas every year. To cope with Variegata wilt stress, plants initiate complex physiological and biochemical responses, including tissue structure modification, accumulation of antimicrobial substances, reactive oxygen species (ROS) bursts and homeostasis, calcium signaling, mitogen-activated protein kinase (MAPK) cascades, hormone signaling, and PAMPs / effective agent-triggered immune responses (PTI / ETI). Furthermore, Variegata wilt stress leads to excessive accumulation of ROS in plants; if not cleared in time, it can damage plant biofilms, thereby affecting normal plant physiological functions. Therefore, to address the serious threat of Variegata wilt to cotton production, breeding superior disease-resistant varieties is the fundamental measure to solve this problem.

[0004] In recent years, global climate change has led to frequent occurrence of Verticillium wilt, and the demand for fiber quality improvement is urgent, which poses a double challenge to cotton breeding. Fiber development involves cell elongation and secondary wall synthesis, while Verticillium wilt resistance depends on the accumulation of secondary metabolites (such as lignin and flavonoids) and the activation of immune signaling pathways. However, current researches mainly focus on single trait regulatory genes, which is difficult to break through the bottleneck of "high yield but not resistant to disease, and resistant to disease but not high yield". Therefore, it is of great significance to excavate key genes that simultaneously regulate fiber development and disease resistance and to analyze their synergistic mechanisms for realizing the coordinated improvement of multiple traits. SUMMARY

[0005] In order to overcome the shortcomings of the prior art, the purpose of the present application is to provide a method for synergistically regulating cotton fiber elongation and Verticillium wilt resistance in Gossypium hirsutum GhMP The method for synergistically regulating cotton fiber elongation and Verticillium wilt resistance in Gossypium hirsutum GhMP The method for synergistically regulating cotton fiber elongation and Verticillium wilt resistance in Gossypium hirsutum

[0006] To achieve the above-mentioned purpose, the present application provides the following scheme: A method for synergistically regulating cotton fiber elongation and Verticillium wilt resistance in Gossypium hirsutum GhMP The method for synergistically regulating cotton fiber elongation and Verticillium wilt resistance in Gossypium hirsutum S1, constructing an expression regulation vector for regulating the expression level of the target gene; GhMP S2, introducing the expression regulation vector into a cotton plant to obtain a transgenic plant with up-regulated or down-regulated expression of the target gene; S3, verifying the transcription level of the plant obtained in step S2 to determine the expression change relative to the wild type; GhMP S4, performing phenotype determination under fiber development and pathogen stress evaluation conditions: when the expression is increased, the cotton fiber length of the target plant is increased and the Verticillium wilt resistance is improved; when the expression is decreased, the cotton fiber length of the target plant is reduced and the Verticillium wilt resistance is reduced. GhMP Preferably, the open reading frame of the target gene has a length of 1695 bp and encodes 564 amino acids. GhMP Preferably, the expression regulation vector of step S1 comprises: GhMP an overexpression vector containing the coding sequence of the target gene; and / or an RNA interference vector containing the coding sequence of the specific segment sequence of the target gene.

[0007] GhMP Preferably, the overexpression vector contains the coding sequence of the target gene.

[0008] Preferably, the RNA interference vector contains the coding sequence of the specific segment sequence of the target gene. GhMP Preferably, the overexpression vector contains the coding sequence of the target gene. GhMP Preferably, the RNA interference vector contains the coding sequence of the specific segment sequence of the target gene.

[0009] ​​​​Preferably, step S2 is introduced via Agrobacterium-mediated genetic transformation.

[0010] Preferably, the transcriptional level verification in step S3 is performed using real-time quantitative PCR.

[0011] Preferably, the phenotypic assessment of fiber length is carried out during the fiber development stage, 5–20 days after flowering, and compared with the wild-type control.

[0012] Preferably, the phenotypic assessment of Verticillium wilt resistance is performed 14–30 days after pathogen inoculation and compared with a wild-type control.

[0013] Preferably, GhMP Proteins are located in the cell membrane and interact with it. GhPIP2.7 and GhTIP1.1 The interaction occurred, and the interaction was verified by yeast two-hybrid, bimolecular fluorescence complementation, and dual luciferase complementation assays.

[0014] Preferably, GhMP , GhPIP2.7 and GhTIP1.1 It participates in the transmembrane transport of hydrogen peroxide, and its function was verified by expressing the protein in yeast cells and growing them under hydrogen peroxide treatment conditions.

[0015] A transgenic cotton plant, obtained by the above method, comprising components for regulating... GhMP The expression box of expression, making GhMP The expression changed relative to the corresponding wild type, and compared with the corresponding wild type, it showed differences in cotton fiber length and Verticillium wilt resistance. GhMP Changes in expression correspond to corresponding synergistic changes.

[0016] According to specific embodiments provided by the present invention, the present invention discloses the following technical effects: This invention has linked a key gene controlling fiber length in upland cotton through genome-wide association analysis. GhMP This gene is also upregulated by Verticillium wilt. This invention, through overexpression and RNAi transgenic cotton, creatively discovered… GhMP Its synergistic function in regulating cotton fiber elongation and Verticillium wilt resistance provides important genetic resources for molecular breeding of high-quality, disease-resistant cotton.

[0017] This invention provides a new approach for breeding high-quality, disease-resistant cotton plants by overexpression, enabling the production of cotton plants with increased fiber elongation and resistance to Verticillium wilt. Attached Figure Description

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments will be briefly introduced as follows. Obviously, the drawings described in the following embodiments are only some embodiments of the present application, and all other drawings obtained by those of ordinary skill in the art without creative labor based on these drawings also belong to the protection scope of the present application.

[0019] Figure 1 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; Figure 2 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; GhMP Schematic diagram of being located on the cell membrane; Figure 3 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; GhMP Schematic diagram of transcription level analysis of overexpression and RNAi strain in fiber and leaf tissue; Figure 4 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; GhMP Schematic diagram of positively regulating fiber elongation; Figure 5 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; GhMP Schematic diagram of positively regulating Verticillium wilt resistance.

[0020] Figure 6 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application; GhMP Schematic diagram of interacting with GhPIP2.7 , GhTIP1.1 .

[0021] Figure 7 GWAS association analysis, expression pattern and gene bioinformatics analysis schematic diagram provided for the embodiments of the present application GhMP Schematic diagram of participating in hydrogen peroxide transport with GhPIP2.7 , GhTIP1.1 . DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the present application will be described clearly and completely in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only some embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor also belong to the protection scope of the present application.

[0023] The purpose of the present application is to provide a Gossypium hirsutum GhMP application of the gene in synergistically regulating cotton fiber elongation and Verticillium wilt resistance, by regulating the expression level of the Gossypium hirsutum GhMP gene, the positive regulation of cotton fiber length and Verticillium wilt resistance is realized, and key gene resources and technical means are provided for molecular breeding of high-quality cotton varieties.

[0024] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0025] This embodiment first performed a genome-wide association analysis on 419 upland cotton accessions, discovering 212 SNPs significantly associated with fiber length (FL) in a candidate region on chromosome Dt11, and identifying the gene for this project. GhMP Furthermore, SV-GWAS of 1081 upland cotton materials was used to further identify structural variations associated with FL at Dt11, one of which was located at... GhMP The gene is located 550 bp upstream. This was determined through cloning of varieties with differences in fiber length. GhMP The gene promoter sequence further confirms the presence of this SV site. These data suggest that this gene may affect fiber length. GhMP Encoding a membrane protein, with an open reading frame of 1695 bp, encoding 564 amino acids, and a molecular weight of approximately 62.76 kDa. Conserved domain analysis revealed... GhMP The absence of known conservative structural domains indicates GhMP It is a novel gene, potentially possessing unique biological functions or molecular mechanisms. Transmembrane domain analysis revealed... GhMP The N-terminus has five transmembrane structures. Subcellular localization analysis indicates... GhMP It is located in the cell membrane. Tissue expression characterization analysis revealed... GhMP The gene is highly expressed in root tissues and differentially expressed after being induced by Verticillium wilt, suggesting that it may play a role in Verticillium wilt resistance.

[0026] To clarify GhMP To investigate the role of this gene in fiber development, this example constructed overexpression and RNAi transgenic lines for this gene. qRT-PCR analysis showed that, at 15 days post-flowering (10⁵ DPA) in fibers, the overexpression lines... GhMP Transcriptional levels were significantly increased by 20-40 times compared to wild-type (WT), while the interference efficiency of this gene in RNAi lines was around 60%. Fiber dynamic elongation analysis showed that during fiber development at 5-20 DPA and into mature fibers, the fiber length of overexpressing lines was consistently higher than that of wild-type, while the fiber length of RNAi lines was significantly lower than that of wild-type. This confirms... GhMP Positive regulation of fiber elongation.

[0027] Further clarification GhMP To investigate the role of this gene in the response to Verticillium wilt, this study utilized overexpression of the gene and RNAi transgenic lines. qRT-PCR analysis showed that in the first true leaf, the overexpressing lines... GhMPThe transcription level was significantly increased by 7-10 times compared with the wild type (WT), and the interference efficiency of the gene in the RNAi strain was about 70%. The resistance response after Verticillium dahliae infection was continuously observed, and the disease index of the overexpression strain was continuously lower than that of the wild type 17-30 days after inoculation, while the disease index of the RNAi strain was continuously higher than that of the wild type.

[0028] In order to further explore the molecular function of GhMP , the CDS sequence of GhMP was connected to the pGBKT7 vector as a bait to screen the interaction proteins by yeast two-hybrid (Y2H) with the cotton fiber yeast library as prey. Two candidate interaction proteins were identified by screening, named GhPIP2.7 and GhTIP1.1 (both encoding aquaporins). In order to verify the interaction between GhMP and GhPIP2.7 , GhTIP1.1 , this example used double-molecule fluorescence complementation (BiFC), double-fluorescent luciferase complementation (LUC) and other experiments to verify, which proved that there was interaction between GhMP and GhPIP2.7 , GhTIP1.1 , indicating that they may form a regulatory complex to play a role in cotton fiber development and Verticillium wilt response.

[0029] Aquaporins are tetrameric channel proteins that transport small molecules (including H202) across membranes under growth and development and related stress, maintaining intracellular homeostasis. In order to explore the effect of GhMP on hydrogen peroxide, the CDS sequences of GhMP , GhPIP2.7 and GhTIP1.1 were connected to the pYES2 vector and expressed in yeast strain BY4742. By observing the growth of yeast cells under hydrogen peroxide treatment, it was shown that GhMP and GhPIP2.7 , GhTIP1.1 were involved in hydrogen peroxide transport.

[0030] The biological materials used in this example are introduced as follows: Escherichia coli Trans1-T1 was purchased from Beijing Zison Bio-technology Co., Ltd., with the item number CD501; Agrobacterium GV3101 was purchased from Shanghai Weidi Biological Technology Co., Ltd., with the item number AC1001; pGreen-GFP vector was used for transient expression of green fluorescent protein in tobacco, pGBKT7 and pGADT7 vectors were used for yeast two-hybrid verification test, pGreenII0800-Luc vector was used for transient expression of LUC in tobacco, and pYES2 vector was used for expression of proteins in yeast cells, all of which were preserved in the laboratory. The cotton germplasm materials used were collected and preserved by the laboratory. The nucleotide sequence of the gene or the amino acid sequence of the protein used in this example is as follows: GhMP Coding sequence: GhMP amino acid sequence: MDAGNSNQAGWLRIVYVVFAFCSALFLGALKGLLVGPIAALILIIGNLGVILGLLPAHIAWTIYTVVKTNRFDAPLKVALLIALPALFGIWLGLSIAGTVIVSVCYGFFTPWVSSFEAFRLDDESDRFFHCVVDGTWDTIK GSCTVVRDFSDLCFHSYPLYLKELRESPVSNEVRTLRLIHVPGLIVVGLGLIVHIPIYTIIAIVKSPYMLFRGWFRLTHDLISREGPFLETACIPVAGLTILLWPIVVIGSIIMAIFSSIFIGLYGSVIVYQERSFKRGV AYVIAMIAEFDEYTNDWLYLREGTIFPKPSYRKKNGSEIEYSVGGLGGRFSSTTGEPPAMLMPTLARSVREAIKEVKMVQVWTNVMKSCEIRGKELLEAKVITSLDLCEWLKAKGSNEGAIIGVGLPCYSFLQTLLVSI RSGSNGLLMLDNVEINSLNRPKDKLLDWFFNPIMVLKEQIRVIKLGDGEVKLLEKLVLFGTNLERMDAWDNGSIVPQDSLRAAQMEGISRRMIGIARSISKLPTYRRKFRQVVKELITHASDKQDIPKCGSIKSTSSYEQV Example 1 GhMP Related to fiber elongation and resistance to Verticillium wilt Based on SNP-GWAS analysis of 419 upland cotton samples and fiber length traits and SV-GWAS analysis of 1081 upland cotton samples, this invention discovered that the trait located on chromosome D11... GhMP The gene is significantly associated with cotton fiber length. Notably, one of the structural variation sites is located at... GhMP At a position 550 bp upstream of the gene, differences in fiber length were observed in cloning varieties. GhMP The gene promoter sequence further confirms the existence of this SV site. Figure 1 A). These data suggest that this gene may influence fiber length. (Based on measurements...) GhMP The expression levels of genes at different developmental stages in upland cotton varieties with differences in fiber quality were investigated. GhMPThe expression level was higher in high-quality fiber varieties than in low-quality upland cotton varieties, mainly in fibers at 0 days after flowering (0 DPA) and after 15 days after flowering (>15 DPA). Figure 1 B), indicating GhMP It may play a key role in fiber initiation and secondary wall thickening. Tissue expression characterization analysis revealed... GhMP High expression in root tissue ( Figure 1 C), and differential expression after Verticillium wilt induction, as evidenced by a significant increase in gene expression after Verticillium wilt infection (C). Figure 1 (D) indicates that this gene is likely involved in the resistance response to Verticillium wilt.

[0031] GhMP It encodes a membrane protein with an open reading frame of 1695 bp, encoding 564 amino acids, and a molecular weight of approximately 62.76 kDa. Analysis of the protein's conserved domains revealed... GhMP No known conservative structural domains ( Figure 1 E), indicating GhMP It is a novel gene, potentially possessing unique biological functions or molecular mechanisms. Based on its encoding of a membrane protein, transmembrane domain analysis revealed… GhMP It has 5 transmembrane domains at its N-terminus ( Figure 1 F). Subcellular localization analysis showed GhMP Located in the cell membrane ( Figure 2 ). Foreshadowing GhMP It may integrate molecular networks of fiber elongation and disease resistance response through transmembrane transport and metabolic remodeling.

[0032] Example 2 GhMP Positive regulation of fiber elongation To clarify GhMP To investigate the role of this gene in fiber development, this example constructed overexpression and RNAi transgenic lines for this gene. qRT-PCR analysis showed that, at 15 days post-flowering (10⁵ DPA) in fibers, the overexpression lines... GhMP Transcription levels were significantly increased by 20-40 times compared to wild-type (WT) (e.g. Figure 3 A), while the interference efficiency of this gene in RNAi lines is around 60% ( Figure 3 B). To clarify GhMP The role of transgenic cotton in fiber elongation was dynamically observed at 5, 10, 15, and 20 DPA after flowering. Results showed that during the 5-20 DPA period, the fiber length of the transgenic lines was consistently longer than that of the wild type, while the fiber length of the RNAi transgenic lines was significantly shorter than that of the wild type. This confirms... GhMP Positive regulation of fiber elongation ( Figure 4 ).

[0033] Example 3 GhMP Positive regulation of cotton Verticillium wilt resistance To further clarify GhMP The role of overexpression and RNAi transgenic lines in Verticillium wilt resistance response was investigated by inoculating them with Verticillium wilt pathogen. qRT-PCR analysis showed that in the first true leaf, the overexpression lines... GhMP Transcription levels were significantly increased by 7-10 times compared to wild-type (WT). Figure 3 C), while the interference efficiency of this gene in RNAi lines is around 70% ( Figure 3 D). To clarify GhMP The role of overexpression in Verticillium wilt resistance response was investigated by continuously observing resistance performance after Verticillium wilt infection. From 17 to 30 days post-infection, the disease index of overexpression lines remained consistently lower than that of wild-type, while the disease index of RNAi lines remained consistently higher than that of wild-type. Figure 5 ).

[0034] Example 4 GhMP and GhPIP2.7 , GhTIP1.1 Interactions affecting hydrogen peroxide transport jointly regulate fiber elongation and Verticillium wilt resistance To explore further GhMP The molecular function, in this embodiment will GhMP The CDS sequence was ligated into the pGBKT7 vector as bait, and yeast coryza libraries were used as prey for screening interacting proteins in yeast two-hybrid (Y2H) experiments. Two candidate interacting proteins were identified and named... GhPIP2.7 and GhTIP1.1 (Both encode aquaporins). To verify GhMP and GhPIP2.7 , GhTIP1.1 The interaction between them will GhPIP2.7 and GhTIP1.1 The CDS sequence was ligated to the pGADT7 vector, and then ligated to the CDS sequence ligated to the pGBKT7 vector. GhMP The recombinant vector was co-transformed into Y2H-gold cells and cultured on SD / -Leu / -Trp / -His / -Ade medium. Results showed colony growth on the positive control plate, indicating the effectiveness of the yeast experimental system, while no colony growth was observed on the corresponding negative control plate. pGBKT7- GhMP +pGADT7- GhPIP2.7 and pGBKT7- GhMP +pGADT7- GhTIP1.1 The presence of bacterial colony growth indicates GhMP and GhPIP1.1, GhTIP1.1 There is an interaction relationship between them. Figure 6 A, B). To further verify GhMP andGhPIP2.7 , GhTIP1.1 Whether the interaction between GmNMY2 and GmNMY3 exists in plant body, this example carried out BiFC experiment in N. benthamiana leaves, the results showed that when GmNMY2-nYFP and GmNMY3-cYFP fusion proteins or GmNMY3-nYFP and GmNMY2-cYFP fusion proteins were co-expressed in the lower epidermis cells of tobacco leaves, YFP fluorescence could be specifically observed in the cell membrane (Fig. C, D). In addition, similar results were also obtained by Dual-Luciferase complemententation Assay (LUC) (Fig. E, F). Summarizing the results of Y2H, BiFC and LUC experiments, it was shown that the interaction between GmNMY2 and GmNMY3 exists in the cell membrane, and they may form a regulatory complex to jointly play a role in cotton fiber development and Verticillium wilt response. GhMP GhPIP2.7 GhTIP1.1 Figure 6 C, D). In addition, similar results were also obtained by Dual-Luciferase complemententation Assay (LUC) (Fig. E, F). Summarizing the results of Y2H, BiFC and LUC experiments, it was shown that the interaction between GmNMY2 and GmNMY3 exists in the cell membrane, and they may form a regulatory complex to jointly play a role in cotton fiber development and Verticillium wilt response. Figure 6 GhMP GhPIP2.7 , GhTIP1.1

[0035] Aquaporins are tetrameric channel proteins that transport small molecules (including H2O2) across the membrane under growth and development and related stresses, maintaining the homeostasis of intracellular environment. To verify the influence of GmNMY2, GmNMY3 and GmNMY4 on hydrogen peroxide, the CDS sequences of GmNMY2, GmNMY3 and GmNMY4 were connected to pYES2 vector and expressed in yeast strain BY4742. In yeast growth experiment, the yeast transformants were diluted to specific OD 600 values (0.70, 0.07 and 0.007) and uniformly spotted on SD-URA+GAL solid medium containing 0 or 1.5 mmol / L H2O2. After 3 d culture at 30℃, the viability of the yeast transformants was recorded. The results showed that the growth of the yeast transformants of GmNMY2, GmNMY3 and GmNMY4 on the medium containing H2O2 was inhibited, and the growth curves determined by liquid culture also proved the results (Fig. ). GhMP GhMP , GhPIP2.7 GhTIP1.1 GhMP GhPIP2.7 GhTIP1.1 The above results showed that GmNMY2, GmNMY3 and GmNMY4 have the ability to transport H2O2. Figure 7 GhMP GhPIP2.7 GhTIP1.1

[0036] The various embodiments in the specification are described in a progressive manner, and each embodiment focuses on the difference from other embodiments. The same and similar parts between the various embodiments can be referred to each other.

[0037] ​​​​​​​​​​​​​​​The principles and implementation manners of the present application are described by using specific examples in the present application, and the above examples are only used to help understand the method of the present application and its core idea; meanwhile, for the general technical personnel in the art, the specific implementation manners and application ranges will be changed according to the idea of the present application. In conclusion, the content of the present specification should not be understood as the limitation of the present application.

Claims

1. A method based on upland cotton GhMP A method for gene-coordinated regulation of cotton fiber elongation and Verticillium wilt resistance, characterized in that, comprising: S1, constructing an expression regulatory vector for regulating GhMP expression level of the expression regulatory vector; S2, introducing the expression regulation vector into the cotton plant to obtain GhMP transgenic plants in which expression is up-regulated or down-regulated S3. The plants obtained in step S2 GhMP The transcriptional level was validated to determine the expression changes relative to the wild type; S4. Phenotyping under fiber development and pathogen stress evaluation conditions: when GhMP When expression is increased, the target plant has increased cotton fiber length and increased resistance to Verticillium wilt. When GhMP When expression is reduced, the cotton fiber length of the target plant is reduced and the resistance to Verticillium wilt is reduced.

2. The method of claim 1, wherein, GhMP The open reading frame length is 1695 bp, encoding 564 amino acids.

3. The method of claim 1, wherein, The expression regulatory vector of step S1 comprises: comprising GhMP overexpression vectors for coding sequences; and / or containing GhMP RNA interference vectors encoding specific segment sequences.

4. The method of claim 1, wherein, The introduction method of step S2 is Agrobacterium-mediated genetic transformation.

5. The method of claim 1, wherein, The transcription level verification of step S3 adopts real-time fluorescent quantitative PCR.

6. The method of claim 1, wherein, The phenotype evaluation of fiber length is performed at the fiber development stage of 5-20 days after flowering, and compared with the wild type control.

7. The method of claim 1, wherein, The phenotype evaluation of yellow wilt resistance is performed at 14-30 days after pathogenic inoculation, and compared with the wild type control.

8. The method of claim 1, wherein, GhMP Protein is located in the cell membrane and interacts with GhPIP2.7 with GhTIP1.1 interactions were verified by yeast two-hybrid, bimolecular fluorescence complementation and dual luciferase complementation assays.

9. The method of claim 8, wherein, GhMP 、 GhPIP2.7 With GhTIP1.1 Involved in the transport of hydrogen peroxide across the membrane and functionally validated by expressing the protein in yeast cells and applying growth conditions under treatment with hydrogen peroxide.

10. A transgenic cotton plant, characterized in that, The plant obtained from the method of claim 1 comprises a cassette for modulating GhMP expression, such that GhMP expression is altered relative to the corresponding wild type and exhibits a synergistic change in cotton fiber length and Verticillium wilt resistance relative to the corresponding wild type. GhMP corresponding to the change in expression.

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