Transcription factor for regulating and controlling drug resistance of plant pathogenic fungi and application of transcription factor

Abstract

The invention discloses a function of a zinc finger transcription factor in regulating resistance of plant pathogenic fungi to various bactericides, which is a transcription factor with a zinc finger structure obtained from botrytis cinerea and named as Bcmr1. After the transcription factor is over-expressed in a botrytis cinerea standard strain B05.10, the sensitivity of pathogenic bacteria to bactericides with various action mechanisms is reduced, and the sensitivity of the pathogenic bacteria to the bactericides after deletion of the transcription factor is remarkably improved. The invention provides a synergist screening candidate target for the sensitivity recovery of the phytopathogen with multidrug resistance in the field to the bactericide, and is beneficial to the treatment of the multidrug resistance of the phytopathogen and the improvement of the activity of the existing bactericide.

Description

Technical Field

[0001] The invention belongs to the field of molecular biology, and more specifically, relates to a transcription factor for regulating the drug resistance of plant pathogenic fungi and its application. Background Art

[0002] Botrytis cinerea is an important fruit and vegetable disease that can cause serious crop yield reductions. Chemical fungicides are often used for disease control in agricultural production, but its pathogen Botrytis cinerea is prone to drug resistance or even multidrug resistance. At present, the resistance control of botrytis cinerea mainly relies on the rotation or combination of agents with different mechanisms of action. Multidrug resistance is mainly caused by the overexpression of ABC efflux proteins related to drug efflux in pathogens, involving the generation of resistance to multiple types of fungicides with different mechanisms of action. There is currently no effective control method in the field, which is a global problem in the field of plant protection. Summary of the invention

[0003] Therefore, the purpose of the present invention is to provide a transcription factor and its encoding gene and application for regulating multidrug resistance of plant pathogenic fungi. The transcription factor regulates the expression of multiple downstream ABC efflux protein encoding genes, thereby affecting the level of drug resistance, providing a target protein for the development of synergists for multidrug resistance management and activity enhancement of existing fungicides, which is a new resistance management strategy.

[0004] The above technical purpose of the present invention is achieved through the following technical scheme: a transcription factor that regulates the drug resistance of plant pathogenic fungi, the transcription factor is the Bcmr1 transcription factor, the transcription factor includes a Zn(II)2Cys6 zinc finger structure, and the amino acid sequence of the zinc finger domain is the amino acid residue sequence at positions 70-103 of the amino terminal of SEQ ID No. 2 in the sequence list.

[0005] The transcription factors are:

[0006] 1) the protein as shown in SEQ ID No. 2;

[0007] 2) A protein derived from the protein shown in SEQ ID No. 2 having the same function as that obtained by replacing and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID No. 2.

[0008] The coding gene of the transcription factor is as follows B1) or B2) or B3):

[0009] B1) a DNA molecule represented by the nucleotide sequence of SEQ ID NO.1 in the sequence list;

[0010] B2) a cDNA molecule or a DNA molecule having more than 95% identity with the nucleotide sequence shown in B1) and encoding the transcription factor;

[0011] B3) A cDNA molecule or a DNA molecule which hybridizes with the nucleotide sequence defined in B1) or B2) under stringent conditions and encodes the transcription factor described in claim 1.

[0012] The present invention also provides a biological material containing the nucleic acid molecule encoding the gene, which is any one of the following C1) to C10):

[0013] C1) an expression cassette containing the encoding gene;

[0014] C2) a recombinant vector containing the coding gene, or a recombinant vector containing the expression cassette described in C1);

[0015] C3) a recombinant microorganism containing the encoding gene, or a recombinant microorganism containing the expression cassette described in C1), or a recombinant microorganism containing the recombinant vector described in C2);

[0016] C4) a transgenic plant cell line containing the coding gene of claim 3, or a transgenic plant cell line containing the expression cassette of C1);

[0017] C5) transgenic plant tissue containing the coding gene of claim 3, or transgenic plant tissue containing the expression cassette of C2);

[0018] C6) a transgenic plant organ containing the coding gene of claim 3, or a transgenic plant organ containing the expression cassette of C2);

[0019] C7) a nucleic acid molecule that inhibits the expression of the gene encoding the gene according to claim 3;

[0020] C8) an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line containing the nucleic acid molecule described in C7);

[0021] C9) a nucleic acid molecule that inhibits the translation of the RNA molecule according to claim 3;

[0022] C10) An expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line for producing the nucleic acid molecule described in C9).

[0023] The use of the transcription factor of the present invention or the encoding gene in regulating the drug resistance of plant pathogenic fungi also falls within the protection scope of the present invention.

[0024] Wherein, the plant pathogen is Botrytis cinerea.

[0025] The use of the transcription factor or its encoding gene in regulating the expression of ABC efflux protein genes (such as BcatrA, BcatrD, BcatrO, Bmr3, BCIN_03g02050 and BCIN_16g02970) in Botrytis cinerea also falls within the protection scope of the present invention.

[0026] The use of the transcription factor or its encoding gene or the biological material in regulating the survival fitness of Botrytis cinerea also falls within the protection scope of the present invention.

[0027] The survival fitness is the adaptability to H2O2 treatment and potassium chloride.

[0028] The application of the transcription factor-related biological materials in regulating the drug resistance of plant pathogenic fungi also falls within the protection scope of the present invention.

[0029] The present invention also protects a method for regulating the drug resistance of plant pathogenic fungi, comprising the following steps:

[0030] 1) transferring the coding gene of the transcription factor according to claim 3 into a plant pathogenic fungus for expression, thereby reducing the sensitivity of the plant pathogenic fungus to pesticides;

[0031] or,

[0032] 2) inactivating the gene encoding the transcription factor according to claim 3 in plant pathogenic fungi, thereby increasing the sensitivity of the plant pathogenic fungi to pesticides;

[0033] The pesticide is a fungicide, preferably one or more selected from microtubule inhibitors, signal transduction inhibitors, DMI fungicides, QoI fungicides, SDHI fungicides and histone inhibitors;

[0034] The plant pathogenic fungus is Botrytis cinerea;

[0035] The microtubule protein inhibitor is one or more of carbendazim, thiabendazim and methylthiophanate;

[0036] The signal transduction inhibitor is one or more of fludioxonil, procymidone and iprodione;

[0037] The DMI fungicide is one or more of difenoconazole, prochloraz and tebuconazole;

[0038] The QoI fungicide is a methoxyacrylate fungicide, preferably one or more of azoxystrobin, pyraclostrobin and trifloxystrobin;

[0039] The SDHI fungicide is a succinate dehydrogenase inhibitor, specifically one or more of boscalid and fluopyrabdo;

[0040] The histone inhibitor is pyrimethanil.

[0041] In summary, the present invention has the following beneficial effects: In the present invention, the transcription factor can activate the expression of downstream drug resistance genes in pathogens by binding to the promoter region of the ABC efflux protein gene, thereby initiating the drug resistance mechanism of the pathogens. Knocking out or inhibiting a single drug metabolism and detoxification-related gene such as the ABC efflux protein encoding gene in pathogens can generally only reduce the resistance of pathogens to a certain fungicide to a low degree, while targeting a key transcription factor can inhibit downstream efflux genes, thereby reducing the resistance of pathogens to drugs with multiple mechanisms of action. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is a schematic diagram of the transcription factor Bcmr1 domain in Example 1 of the present invention

[0043] Figure 2 pXEH-eGFP overexpression vector plasmid map

[0044] Figure 3 PCR verification of Bcmr1 overexpression vector

[0045] Figure 4 pXEH knockout vector plasmid map

[0046] Figure 5 PCR verification of Bcmr1 knockout vector

[0047] Figure 6 PCR verification of Bcmr1 overexpression transformants

[0048] Figure 7 PCR verification of Bcmr1 knockout transformants

[0049] Figure 8 is the relative expression level of Bcmr1 gene in Bcmr1 knockout and overexpression transformants

[0050] Fig. 9 This is the vector plasmid map of Bcmr1 exogenous expression protein

[0051] Fig.10 Gel chromatography retardation method was used to analyze the binding of Bcmr1 exogenous expression protein to its target gene BcatrD promoter

[0052] Fig.11 The relative expression of the downstream target gene BcatrD in Bcmr1 knockout and overexpression transformants

[0053] Fig.12 Survival fitness test of mutant strains

[0054] Fig.13 Determination of relative expression of transcription factor Bcmr1 in resistant mutant strains DETAILED DESCRIPTION

[0055] The methods in the following embodiments are conventional methods unless otherwise specified.

[0056] Example 1: Cloning and analysis of the transcription factor Bcmr1 that regulates drug resistance in plant pathogenic fungi

[0057] 1. Using the standard strain of Botrytis cinerea B05.10 as the material, a drug-resistant Botrytis cinerea-related gene Bcmr1 was cloned. Its CDS sequence length was 2418 bp, encoding 805 amino acids, and containing a conserved Zn(II)2Cys6 zinc finger structure, such as Figure 1 And as shown in Table 1.

[0058] Table 1. Domains, repeat sequences, motifs and feature predictions:

[0059] Name Start End E-value low complexity 12 39 N / A low complexity 47 59 N / A GAL4 69 112 3.64E-13 low complexity 148 159 N / A low complexity 181 191 N / A low complexity 322 337 N / A Fungal_trans 398 471 3.89E-08

[0060] The method for obtaining the transcription factor Bcmr1 coding gene of the present invention is as follows: using the cDNA of the Botrytis cinerea standard strain B05.10 as a template, performing PCR amplification, and the amplification primers are as follows:

[0061] Bcmr1 forward primer: 5'-ATGGATTATTTCAGTGACTCAAATACGA-3'

[0062] Bcmr1 reverse primer: 5'-TTATAGCCCAATTCCTGCATTTGTAA-3'

[0063] The amplification results are as follows Figure 1 As shown, the CDS coding sequence of the gene is 2418 bp in length, as shown in sequence 1 in the sequence list, and encodes 805 amino acids, as shown in sequence 2 in the sequence list.

[0064] 2. Bcmr1 Homology Comparison

[0065] The homology of Bcmr1 was compared by BLAST program in NCBI database, and it was found that its homology with Mrr2 of Candida albicans was 19.53%. The sequence homology of this transcription factor Bcmr1 and related proteins of the same genus fungi was high, but they were all hypothetical proteins that had not been studied.

[0066] 3. Analysis of Physicochemical Properties of Bcmr1

[0067] Analysis of Bcmr1 revealed that its molecular weight is 90645.20, theoretical isoelectric point is 5.86, atomic composition is C4008H6298N1118O1203S39, instability index is 48.64, and fat index is 81.81. The results of hydrophilicity analysis showed that it is a hydrophobic protein.

[0068] Example 2: Function and application of the transcription factor Bcmr1 of the present invention

[0069] 1. Construction of Bcmr1 overexpression vector and knockout vector

[0070] (1) Construction of Bcmr1 overexpression vector:

[0071] Using the cDNA of Botrytis cinerea strain B05.10 as a template, Bcmr1 forward primer and Bcmr1 reverse primer were used as primers to amplify the Bcmr1 fragment, and the overexpression vector pXEH-eGFP (the original pXEH vector was digested with BamHI and HindIII and then connected to the promoter oliC and eGFP sequence; Wu, Z., Bi, Y., Zhang, J., Gao, T., Li, X., Hao, J., Li, G., Liu, P., Liu, X. (2024). Multidrug resistance of Botrytis cinerea associated with its adaptation to plant secondary metabolites. mBio, 15 (2), e02237-23) was double-digested with SalI and XbaI. Figure 2 ), the Bcmr1 fragment and the pXEH-eGFP vector fragment were connected by homologous recombination, transformed into Escherichia coli, screened for positive clones on kanamycin-resistant plates, and verified by PCR amplification using universal primers on the vector, and a band of similar size to the target gene was obtained ( Figure 3 ). After verification by sequencing, it was named pXEH-eGFP-Bcmr1.

[0072] The amplification primer pair of the transcription factor Bcmr1 overexpression vector is:

[0073] Bcmr1 forward primer:

[0074] 5-CATCACAATAGTCGACATGGATTATTTCAGTGACTCAAAT-3'

[0075] Bcmr1 reverse primer:

[0076] 5'-CCTTGCCCATTCTAGATAGCCCAATTCCTGCATTTGT-3'

[0077] (2) Construction of Bcmr1 knockout vector:

[0078] Using the DNA of Botrytis cinerea strain B05.10 as a template, primers were designed to amplify 1000 bp sequences upstream and downstream of the Bcmr1 gene and the hygromycin fragment. The PCR products were recovered and purified and then sequenced. The three correctly sequenced fragments were connected to the pXEH vector after double digestion with XhoI and SacI using seamless cloning method (Feng, HQ, Li, GH, Du, SW, Yang, S., Li, XQ, de Figueiredo, P., Qin, QM (2017). The septin protein Sep4 facilitates host infection by plant fungal pathogens via mediating initiation of infection structure formation. Environmental Microbiology, 19 (5), 1730-1749) ( Figure 4 ), the ligation product was transformed into Escherichia coli, and positive clones were screened on kanamycin resistance plates. The recombinant plasmid was verified by PCR, and the band size was consistent with the expected result, indicating that the Bcmr1 knockout vector was successfully obtained. Figure 5 The knockout vector verified to be correct was named pXEH-q-Bcmr1.

[0079] The transcription factor Bcmr1 knockout vector amplification primer pair is:

[0080] Bcmr1 upstream forward primer:

[0081] 5-ATTATTATGGAGAAACTCGAGCCCTCATCTCTTCACAACTCGAGC-3'

[0082] Bcmr1 upstream reverse primer:

[0083] 5'-CGAGCCCACCGATTCTGGAGTCGTGACTTCGGG-3'

[0084] Hygromycin fragment forward primer:

[0085] 5'-CTCCAGAATCGGTGGGCTCGCAGATCTTC-3'

[0086] Hygromycin fragment reverse primer:

[0087] 5'-CATTCCAAAATCTAGAGGATCCCCAACTGGTTCC-3'

[0088] Bcmr1 downstream forward primer:

[0089] 5'-ATCCTCTAGATTTTGGAATGATATAGAAACGTTTT

[0090] Bcmr1 downstream forward and reverse primers:

[0091] 5'

[0092] -CTTGCATGCCTGCAGGTCGACTTCGAATCGATGCTAGTAAAGTAGA-3'

[0093] 2. Obtaining mutant strains

[0094] (1) Obtaining overexpression mutant strains:

[0095] The overexpression vector was transferred into the B05.10 strain by Agrobacterium-mediated transformation. The Agrobacterium containing the overexpression vector pXEH-eGFP-Bcmr1 was mixed with the spore suspension of Botrytis cinerea at a ratio of 1:1. After being coated on cellophane for 2 days, it was transferred to the selection medium containing hygromycin and cultured for 2 days. After the cellophane was removed and cultured for 3 days, the transformants were picked for PCR verification. The amplification results are shown in Figure 6 shown.

[0096] (2) Obtaining knockout mutant strains:

[0097] The knockout vector was transferred into the B05.10 strain by Agrobacterium-mediated transformation. The Agrobacterium containing the knockout vector pXEH-q-Bcmr1 was mixed with the suspension of Botrytis cinerea spores at a ratio of 1:1, spread evenly on the cellophane and transferred to the selection medium containing hygromycin after 2 days. After the cellophane was removed and cultured for 3 days, the transformants were picked for PCR verification. Figure 7 shown.

[0098] 3. Detection of gene expression in mutant strains

[0099] qPCR detection of overexpression transformants: qPCR verification was performed using cDNA of transformants as templates and specific primers of Bcmr1 gene. The expression level of Bcmr1 in overexpression transformants was significantly higher than that of parent strains (2.28-10.11 times). Figure 8 As shown, the obtained overexpression transformants can be used for subsequent functional analysis.

[0100] Knockout transformant detection: qPCR verification was performed using the cDNA of the transformant as a template and the specific primers of the Bcmr1 gene. The results are shown in the figure below. The expression of the Bcmr1 gene was not detected in the Bcmr1 knockout mutant (0.025-0.268 times), indicating that the Bcmr1 gene was knocked out. Figure 8 As shown, the knockout transformants obtained can be used for subsequent functional analysis.

[0101] 4. Evaluation of drug sensitivity of mutant strains

[0102] The mycelial growth rate method was used to determine the sensitivity of the parent strain, Bcmr1 overexpression strain and Bcmr1 knockout strain to fluazinam, fludioxonil, boscalid, azoxystrobin, prochloraz and difenoconazole. A 5-mm-diameter bacterial cake was cut from the edge of the colony of the strain after growing for 3-5 days and inoculated on PDA medium with different concentrations of the drugs. Each treatment was repeated 3 times. After culturing at 18°C ​​for 3-5 days, the diameter was measured and the inhibition rate and EC were calculated. 50 (Inhibition rate (%) = [control colony diameter (mm) - treatment colony diameter (mm) / (control colony diameter (mm) - bacterial cake diameter (5mm)] × 100. The logarithm of the concentration of the drug is used as the X-axis, and the probability value of the inhibition rate is the Y-axis. The toxicity regression equation Y = bX + a is calculated, and the effective inhibition concentration is EC 50 ).

[0103] The knockout transformants were found to have EC values ​​of azoxystrobin, prochloraz, and difenoconazole. 50 Compared with the parent, the EC values ​​of the overexpression transformants to boscalid, azoxystrobin and difenoconazole were significantly lower. 50 Improved, that is, the sensitivity to some drugs increased after knocking out the transcription factor Bcmr1, while the sensitivity decreased after overexpression (Table 2).

[0104] Table 2. Determination of the sensitivity of Bcmr1 overexpression transformants and knockout transformants to fungicides

[0105]

[0106] Among them, CK is the B05.10 strain transformed with the pXEH vector, M1K1, M1K28 and M1K91 are Bcmr1 knockout transformants, and M1O1, M1O2 and M1O4 are Bcmr1 overexpression transformants

[0107] 5. Verification of Bcmr1 protein binding to target gene promoter

[0108] (1) Construction of Bcmr1 exogenous expression vector: amplify Bcmr1 fragment, forward primer F: 5'-AATGGGTCGCGGATCCATGGATTATTTCAGTGACTCAAAT-3'; reverse primer R: 5'-GACGGAGCTCGAATTCTAGCCCAATTCCTGCATTTGT-3'. The purified fragment was ligated into the vector PET-28a after double digestion with BamHI and EcoRI ( Fig. 9 ), verify the correct vector was transferred into E. coli DE3 for expression and protein purification.

[0109] (2) Construction of promoter probe: The forward primer F with FAM (carboxyfluorescein) modification upstream of the gene encoding the ABC efflux protein BcatrD (ABC efflux protein, XP_001555249.2) was designed: 5'-CGATTGTTCGACTACGAACTATCC-3' and the reverse primer R of BcatrD: 5'-GGCCATACTGTAGTGCTGT-3'. The 2000 bp upstream of the promoter was amplified and purified as a probe.

[0110] (3) Gel chromatography blocking: Add 2 μL of DNA fragments labeled with fluorescent probes to a PCR tube, mix with DNA Binding Buffer (pH 7.5 Tris-HCl 50 mM, KCl 100 mM, MgCl2 2.5 mM, DTT 0.2 mM, salmonsperm DNA 2 μg, pH 8.0 glycerol 10%), incubate in the dark for 20 min, then add 2 μL of the protein obtained in (2), incubate in the dark for 15 min; add 5 μL of DNA Loading dye to the system and transfer to a 4% non-denaturing polyacrylamide gel well for loading; place the electrophoresis device on ice, perform electrophoresis at 110 V until the sample reaches the bottom of the gel, remove the gel glass plate and observe at 490 nm. Fig.10 As shown, transcription factors can reduce the probe migration rate, indicating that the transcription factor Bcmr1 can bind to the upstream promoters of BcatrA, BcatrD, BcatrO, Bmr3, BCIN_03g02050 and BCIN_16g02970.

[0111] (4) ChIP-seq detection of Bcmr1 overexpression transformants

[0112] The mycelium of Bcmr1 overexpressing transformants was collected for formaldehyde cross-linking and DNA was interrupted by ultrasonic / enzymatic digestion. Antibodies that specifically bind to the target protein were added to form an antibody-target protein immunoprecipitation complex, and then the cross-linking was removed, the DNA was purified, and the library was constructed for sequencing. The results were analyzed using fastqc, cutadapt, bowtie2, MACS2, HOMER, and R software to obtain target genes that the transcription factor Bcmr1 can bind to, including 27 ABC efflux proteins as shown in Table 3.

[0113] Table 3. ChIP results of Bcmr1 overexpression transformants ABC efflux protein genes

[0114]

[0115]

[0116] (5) Detection of target gene expression in mutant strains

[0117] qPCR detection of Bcmr1 overexpression and knockout transformants: Using cDNA of different types of Bcmr1 transformants as templates, the specific primers of BcatrD gene were used to determine the expression level. Fig.11 As shown, overexpression of the transcription factor Bcmr1 led to the upregulation of the target genes BcatrA, BcatrD, BcatrO, Bmr3, BCIN_03g02050, and BCIN_16g02970, while knockout of the transcription factor Bcmr1 resulted in downregulation of the expression of the target genes BcatrA, BcatrD, BcatrO, Bmr3, BCIN_03g02050, and BCIN_16g02970.

[0118] 6. Survival Fitness Determination of Mutant Strains

[0119] The inhibition rates of the transcription factor Bcmr1 overexpression and knockout mutants and the parent strain B05.10 to 20 mM hydrogen peroxide, 300 μg / mL Congo red, 1 M sorbitol, 0.01% sodium dodecyl sulfate (SDS) and 0.6 M potassium chloride were determined. After hydrogen peroxide treatment, the Bcmr1 knockout mutant showed a significantly higher inhibition than the parent strain, while the inhibition rate of the overexpression mutant was reduced ( Fig.12 a), there was no obvious or regular change between the mutant and the parent strain under the action of Congo red, sorbitol and SDS ( Fig.12 b, 12c and 12d). However, under the action of potassium chloride, the inhibition rate of knockout and overexpression mutants increased compared with the parent strain ( Fig.12 e). Conidia production was significantly reduced in Bcmr1 knockout transformants ( Fig.12f). The sclerotia of Bcmr1 knockout transformants hardly grow, while the sclerotia of overexpression transformants are reduced in number but larger in size than the parent ( Fig.12 g). At the same time, the knockout transformants almost lost their pathogenicity ( Fig.12 h).

[0120] 7. Determination of relative expression of transcription factor Bcmr1 in resistant mutant strains

[0121] qPCR detection of Bcmr1 in mutant strains: The standard strain B05.10, field sensitive strain 223, field resistant strains 309, 242, S91 and 10 secondary metabolite subculture strains were used as cDNA templates, and the specific primers of the Bcmr1 gene were used to determine its expression level. The results are shown in Fig.13 As shown, the transcription factor Bcmr1 was upregulated in various mutant strains, among which the expression was significantly upregulated in the field-resistant strain S91, the secondary metabolites camptothecin, farnesol, flavanone and resveratrol subculture strains.

[0122] The specific embodiments are only explanations of the present invention, and they are not limitations of the present invention. After reading this specification, those skilled in the art can make non-creative modifications to the embodiments as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. A transcription factor for regulating drug resistance of plant pathogenic fungi, characterized in that: The transcription factor comprises a C2H2 zinc finger structure, and the amino acid sequence of the zinc finger domain is the amino acid residue sequence at positions 70 to 103 of the amino terminal of SEQ ID No. 2 in the sequence list.

2. The transcription factor according to claim 1, characterized in that The transcription factors are: 1) the protein shown in SEQ ID No. 2; 2) A protein derived from the protein shown in SEQ ID No. 2 having the same function as that obtained by replacing and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID No.

2.

3. The gene encoding the transcription factor according to claim 1 or 2 is as follows B1) or B2) or B3): B1) a DNA molecule represented by the nucleotide sequence of SEQ ID NO.1 in the sequence list; B2) a cDNA molecule or a DNA molecule having 95% or more identity with the nucleotide sequence shown in B1) and encoding the transcription factor according to claim 1 or 2; B3) A cDNA molecule or a DNA molecule which hybridizes with the nucleotide sequence defined in B1) or B2) under stringent conditions and encodes the transcription factor described in claim 1.

4. The biological material containing the nucleic acid molecule encoding the gene as claimed in claim 3, which is any one of the following C1) to C10): C1) an expression cassette containing the coding gene according to claim 3; C2) a recombinant vector containing the coding gene according to claim 3, or a recombinant vector containing the expression cassette according to C1); C3) a recombinant microorganism containing the coding gene of claim 3, or a recombinant microorganism containing the expression cassette of C1), or a recombinant microorganism containing the recombinant vector of C2); C4) a transgenic plant cell line containing the coding gene of claim 3, or a transgenic plant cell line containing the expression cassette of C1); C5) transgenic plant tissue containing the coding gene of claim 3, or transgenic plant tissue containing the expression cassette of C2); C6) a transgenic plant organ containing the coding gene of claim 3, or a transgenic plant organ containing the expression cassette of C2); C7) a nucleic acid molecule that inhibits the expression of the gene encoding the gene according to claim 3; C8) an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line containing the nucleic acid molecule described in C7); C9) a nucleic acid molecule that inhibits the translation of the RNA molecule according to claim 3; C10) An expression cassette, a recombinant vector, a recombinant microorganism or a transgenic plant cell line for producing the nucleic acid molecule described in C9).

5. The function of the transcription factor shown in claim 1 or 2, the coding gene according to claim 3, or the biological material according to claim 4 in regulating the drug resistance of plant pathogenic fungi.

6. The use according to claim 5, wherein the plant pathogen is Botrytis cinerea; preferably, the drug resistance is resistance to fungicides, and the fungicides are preferably selected from one or more of microtubule inhibitors, signal transduction inhibitors, DMI fungicides, QoI fungicides, SDHI fungicides and histone inhibitors.

7. Use of the transcription factor according to claim 1 or 2 or the coding gene according to claim 3 in regulating the expression of ABC efflux protein genes in Botrytis cinerea.

8. Use of the transcription factor according to claim 1 or 2, the coding gene according to claim 3, or the biological material according to claim 4 in regulating the survival fitness of Botrytis cinerea.

9. The use according to claim 8, wherein the survival fitness is the adaptability to H2O2 treatment and potassium chloride.

10. A method for regulating drug resistance of plant pathogenic fungi, comprising the following steps: 1) transferring the coding gene of the transcription factor according to claim 3 into a plant pathogenic fungus for expression, thereby reducing the sensitivity of the plant pathogenic fungus to pesticides; or, 2) inactivating the gene encoding the transcription factor according to claim 3 in plant pathogenic fungi, thereby increasing the sensitivity of the plant pathogenic fungi to pesticides; The plant pathogenic fungus is Botrytis cinerea; The pesticide is a fungicide, preferably one or more selected from microtubule inhibitors, signal transduction inhibitors, DMI fungicides, QoI fungicides, SDHI fungicides and histone inhibitors; The microtubule protein inhibitor is one or more of carbendazim, thiabendazim and methylthiophanate; The signal transduction inhibitor is one or more of fludioxonil, procymidone and iprodione; The DMI fungicide is one or more of difenoconazole, prochloraz and tebuconazole; The QoI fungicide is a methoxyacrylate fungicide, preferably one or more of azoxystrobin, pyraclostrobin and trifloxystrobin; The SDHI fungicide is a succinate dehydrogenase inhibitor, specifically one or more of boscalid and fluopyrabdo; The histone inhibitor is pyrimethanil.

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