CN114989275B

CN114989275B - Application of OsERF940 protein in improving rice blast resistance

Info

Publication number: CN114989275B
Application number: CN202110147976.1A
Authority: CN
Inventors: 张执金; 张海文; 朱莉; 王嘉翼; 李梓萱; 李小湘
Original assignee: Biotechnology Research Institute of CAAS
Current assignee: Biotechnology Research Institute of CAAS
Priority date: 2021-02-03
Filing date: 2021-02-03
Publication date: 2024-01-02
Anticipated expiration: 2041-02-03
Also published as: CN114989275A

Abstract

The invention discloses an application of OsERF940 protein in improving rice blast resistance. The invention provides an application of any one of the following 1) -3) in regulating and controlling plant rice blast resistance; 1) Protein OsERF940; 2) A nucleic acid molecule encoding the protein OsERF940; 3) A recombinant vector, expression cassette or recombinant bacterium comprising a nucleic acid molecule encoding a protein OsERF940; the protein OsERF940 is a protein composed of an amino acid sequence shown as a sequence 1 in a sequence table. The invention claims the application of OsERF940 protein to increase the expression of rice disease resistance related genes and improve the resistance of rice to rice blast. The invention has great application value for cultivating rice blast resistant rice varieties.

Description

Application of OsERF940 protein in improving rice blast resistance

Technical Field

The invention belongs to the technical field of biology, and particularly relates to application of OsERF940 protein in improving rice blast resistance.

Background

Ethylene response factors (ethylene response factors, ERF) belong to a subfamily of ERF/AP2 superfamily, which share the common feature of containing a conserved ERF domain that regulates plant responses to biotic and abiotic stresses. A large number of genes encoding ERF factors were also isolated from different plants. Related studies around these genes have become a new focus in the field of plant genomics. A great deal of research at home and abroad shows that ERF plays an important role in plant disease resistance response. For example, university of missouri research in the united states indicates that the ERF factor ERF6 plays an important role in the development of resistance to arabidopsis gray mold. The Zhang Zengyan subject group of the national institute of agriculture and crop research found that overexpression of the wheat ERF factor ERF1 can significantly inhibit growth of Rhizoctonia cerealis and improve resistance of wheat to banded sclerotial blight. Studies in yecanada germany have shown that overexpression of the rice ERF factor oseerbbp 1 increases bacterial leaf blight resistance in rice. Research in the Zhang Shuzhen subject group of northeast university of agriculture has found that soybean ERF factor increases soybean resistance to phytophthora root rot. The research at home and abroad shows that the EFR factor has important regulation and control effects on disease resistance of various crops in different crops.

However, the function of ERF gene in Arabidopsis is widely studied, and the function of ERF family genes of most rice and the regulation relationship in disease resistance response are not clear.

Disclosure of Invention

The invention aims to provide an application of OsERF940 protein in improving rice blast resistance.

The invention claims the application of OsERF940 protein to increase rice blast resistance.

The invention also protects the application of the OsERF940 gene in the cultivation of transgenic plants for increasing rice blast resistance.

The invention also provides a plant breeding method, which comprises the following steps: the activity and/or the content of the OsERF940 protein in the target plant is improved, so that the rice blast resistance of the plant is improved.

The invention also provides a method for preparing a transgenic plant, comprising the steps of: the OsERF940 gene is introduced into a starting plant to obtain a transgenic plant with increased rice blast resistance compared with the starting plant.

The OsERF940 protein is specifically derived from the Oryza sativa L.Zhonghua 11 variety in Asian cultivar of Oryza.

Any one of the above OsERF940 proteins is as follows (a) or (b) or (c) or (d) or (e):

(a) A protein shown in a sequence 1 of a sequence table;

(b) A protein derived from the plant rice blast increase-related protein obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence 1;

(c) A protein derived from rice and having 95% or more identity to (a) and being associated with rice blast resistance;

any of the above OsERF940 genes is a gene encoding the OsERF940 protein.

The OsERF940 gene is specifically a DNA molecule as described in (1) or (2) or (3) or (4) below:

(1) A DNA molecule with a coding region shown as a sequence 2 in a sequence table;

(2) A DNA molecule which has 75% or more identity to (1) and which encodes a protein associated with the disease resistance of rice blast in plants;

(3) A DNA molecule derived from rice and having 90% or more identity to (1) and encoding a protein associated with rice blast disease resistance;

(4) A DNA molecule which hybridizes under stringent conditions to (1) and which encodes a protein associated with the resistance of a plant to rice blast;

any of the above plants is a monocotyledonous plant or a dicotyledonous plant. The monocotyledonous plant is a plant of the Gramineae family. The Gramineae plant is a plant of Paddy. The rice plant is rice, such as rice middle flower 11.

In any of the above methods, the "introduction of the OsERF940 gene into the starting plant" is achieved by introducing a recombinant expression vector into the starting plant. The recombinant expression vector is a plasmid which is obtained by inserting the OsERF940 gene into a plant expression vector and can express the OsERF940 gene. The recombinant expression vector can be specifically: the recombinant plasmid pCAMBIA3301-OsERF940 is obtained by inserting double-stranded DNA molecules shown as nucleotide numbers 1-954 of sequence 2 of a sequence table into a multiple cloning site (for example, between Sac I and BamH I cleavage sites) of the pCAMBIA3301 vector.

Experiments prove that the overexpression OsERF940 gene obviously increases the transcriptional expression of rice disease resistance related genes and increases the rice blast resistance. The OsERF940 protein has important function on improving rice blast resistance. The invention has great application value for cultivating rice blast resistant rice varieties.

Drawings

FIG. 1 is a schematic representation of the elements of recombinant plasmid pCAMBIA3301-OsERF940.

FIG. 2 shows the relative expression levels of the OsERF940 gene, M: nucleic acid molecule Marker, control: control (wild type), T1-T8: the RT-PCR detection result of the T0 generation plant line is shown.

FIG. 3 shows qPCR detection of the expression level of the disease-resistant gene in the OsERF940 over-expressed strains OE3 and OE 5.

FIG. 4 is a graph of field and laboratory inoculation of transgenic rice lines with rice blast, where WT represents the wild type control, OE3, OE4, OE5 represent the different transgenic lines; (A) Representing the disease resistance phenotype of the transgenic material in a disease spectrum field; (B) The disease resistance phenotype of the transgenic material under indoor inoculation condition is shown.

Detailed Description

The experimental methods used in the following examples are conventional methods unless otherwise specified.

Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

The following examples facilitate a better understanding of the present invention, but are not intended to limit the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores. The quantitative tests in the following examples were all set up in triplicate and the results averaged.

The protein shown in the sequence 1 in the sequence table is named as OsERF940 protein, the coding region of the OsERF940 protein in rice cDNA is shown in the sequence 2 in the sequence table, and the protein is named as OsERF940 gene.

The pCAMBIA3301 vector, which is fully called pCAMBIA3301-myc plant overexpression vector: the Shanghai Jiran Biotechnology Co.Ltd, catalog number JR13080311.

Agrobacterium LBA4404: the catalogue number of the popular biotechnology company is AABV02-03.

The rice flower 11 is described in the following literature: kwon CT, kimsh, kimd, paek nc.the Rice Floral Repressor Early flowering1 Affects Spikelet Fertility By Modulating Gibberellin signaling.rice (N Y) 2015Dec;8 (1) 58.doi:10.1186/s12284-015-0058-1, available to the public from the applicant.

Pyricularia oryzae strain (strain GD 08-T13), hunan province national institute of sciences, rice institute Li Xiaoxiang gift; is described in the following documents: liu Q, li X, yan S, yu T, yang J, dong J, zhang S, zhao J, yang T, mao X, zhu X, liu B.OsWRKY67 positively regulates blast and bacteria blight resistance by direct activation of PR genes in rice. BMC Plant biol.2018Oct26;18 (1) 257.doi:10.1186/s12870-018-1479-y, available to the public from the applicant.

Example 1 obtaining transgenic plants

1. Construction of recombinant plasmids

1. Extracting total RNA of rice Japanese sunny, and then carrying out reverse transcription to obtain cDNA.

2. And (3) taking the cDNA obtained in the step (1) as a template, adopting a primer pair consisting of F1 and R1 to carry out PCR amplification, and recovering a PCR amplification product of 236 bp.

F1：5'-CGGTAGGATCCATGGTGCCCTCTTCACGGAAAG-3'

R1：5'-CCCCGAGCTCAAGACAGTTGAGTTCTTGTTCCA-3'。

3. And (3) double-enzyme cutting the PCR amplification product obtained in the step (2) by using restriction enzymes Sac I and BamH, and recovering the enzyme-cut product.

4. The pCAMBIA3301 vector was digested with the restriction enzymes SacI and BamH to recover the vector backbone.

5. And (3) connecting the enzyme digestion product obtained in the step (3) with the vector skeleton obtained in the step (4) to obtain the recombinant plasmid pCAMBIA3301-OsERF940.

According to the sequencing result, the recombinant plasmid pCAMBIA3301-OsERF940 is a plasmid obtained by inserting a DNA molecule shown in the 1 st to 951 st positions of the sequence 2 of a sequence table between Sac I and BamH restriction sites of a pCAMBIA3301 vector, and the plasmid contains the coding sequence of an OsERF940 gene and expresses the protein OsERF940.

The schematic diagram of the elements of the recombinant plasmid pCAMBIA3301-OsERF940 is shown in FIG. 1.

2. Obtaining of OsERF940 Gene-transferred Rice

1. Recombinant plasmid pCAMBIA3301-OsERF940 is introduced into Agrobacterium LBA4404 to obtain recombinant Agrobacterium.

2. Suspending the recombinant Agrobacterium obtained in step 1 in liquid ASAA medium to obtain OD _600nm Value = 0.3 bacterial liquid.

Liquid ASAA medium: liquid NB medium containing 2 mg/L2, 4-D and 100. Mu.M/L AS.

3. Seeds of flower 11 (hereinafter referred to as wild type rice) of rice were taken, sterilized with 75% ethanol, then placed on a solid NB medium plate, dark-cultured at 28℃for 2 weeks, then callus was transferred to a new solid NB medium plate for 2 weeks at 28℃and then transferred to a new solid NB medium plate for 2 weeks at 28 ℃.

4. After the step 3 is completed, the embryogenic callus particles which are naturally dispersed in the callus and have light yellow color are placed on a subculture medium plate and are dark-cultured for 3 days at 28 ℃.

Subculture medium: solid NB medium containing 2 mg/L2, 4-D.

5. And (3) after the step (4) is completed, taking the callus, and soaking the callus in the bacterial liquid obtained in the step (2) for 10-20 minutes, and slightly shaking the callus.

6. After the step 5 is completed, the callus is taken, the surface bacterial liquid is sucked by filter paper, and then the callus is placed on a co-culture medium plate for dark culture for 3 days at the temperature of 21-22 ℃.

Co-culture medium: solid NB medium containing 2 mg/L2, 4-D and 100. Mu.M AS.

7. After the step 6 is completed, the callus is taken and placed on a screening medium plate for dark culture at 28 ℃ for 2 weeks, then the callus is transferred to a new screening medium plate for dark culture at 28 ℃ for 2 weeks, and then the callus is transferred to the new screening medium plate for dark culture at 28 ℃ for 2 weeks.

Screening Medium solid NB medium containing 2 mg/L2, 4-D and 50mg/L Bar.

8. And 7, after the step 7 is completed, taking fresh calli which grow vigorously and are milky white or yellowish, placing the calli on a pre-differentiation culture medium plate, carrying out dark culture at 28 ℃ for 1 week, carrying out light culture at 28 ℃ for 2 weeks, transferring the calli onto a differentiation culture medium plate, and carrying out light culture at 28 ℃ to obtain differentiated seedlings.

Pre-differentiation medium: a solid NB medium containing 5mg/L ABA and 0.5mg/L NAA.

Differentiation medium: a solid NB medium containing 0.5mg/L NAA and 3 mg/L6-BA.

9. After the step 8 is completed, transferring the differentiated seedling with good growth vigor into a triangular flask filled with a solid MS culture medium, culturing for 1-2 weeks by illumination at 28 ℃, transplanting into a greenhouse, and continuously culturing until seeds are harvested, namely T ₁ Seed generation.

10. Will T ₁ The generation seeds are cultivated into plants, namely T ₁ Plants of the generation T ₁ Selfing the plant to obtain T ₂ Seed generation.

12. Will T ₂ The generation seeds are cultivated into plants, namely T ₂ Plants of the generation T ₂ Selfing the plant to obtain T ₃ Seed generation.

13. Will T ₃ The generation seeds are cultivated into plants, namely T ₃ And (5) replacing plants.

For a certain T ₁ For the generation of plants, the T is if two conditions are met ₁ The generation plant is a single copy inserted transgene plant: (1) the T is ₁ The plants of the generation have Bar resistance (2% Basta (company: beijing Hua Vietnam biological product number: W9062-100 ml) is sprayed with 3 leaves and one heart age rice seedlings, and after 1 week of treatment, green rice seedlings are screened as positive transgenic materials); (2) the T is ₁ T obtained by selfing of plant generation ₂ In the generation plants, the number ratio of Bar-resistant plants to non-Bar-resistant plants substantially meets 3:1 (after Basta selection, rice plants in the plant line (about 50 plants)Basta resistant seedlings of (a): basta sensitivity Miao 3:1).

For a certain T ₂ For the generation of plants, the T is if the following three conditions are met ₂ The generation plant and the selfing progeny are a homozygous transgenic line: (1) the T is ₂ The plants of the generation have Bar resistance (3 leaves and one heart age rice seedlings are sprayed with 2% Basta in the field, and the rice seedlings remain green after being treated for 1 week); (2) t of it ₁ The generation plants are single copy inserted transgenic plants (Basta resistant seedlings of rice plants (about 50 plants) in the strain after Basta selection: basta sensitive Miao 3:1); (3) t of its sampling detection ₃ The plants of the generation were all Bar resistant (3 leaves, one heart age seedlings were sprayed with 2% basta in the field and the seedlings remained green after 1 week of treatment).

3. PCR identification

Randomly taking 2 strains, namely an OE1 strain and an OE2 strain from homozygous transgenic strains obtained in the step two, and aiming at T ₂ And identifying the generation plants. Flower 11 of rice was used as a wild type control for the transgenic line.

Total RNA in plants (4 weeks seedlings) was extracted, reverse transcribed into cDNA, and the expression level of the OsERF940 gene was identified by quantitative PCR using the cDNA as a template.

Primers for qPCR identification of the OsERF940 gene were as follows:

F2：5'-GATGAGATTCCTGTGAGCAAC-3'；

R2：5'-GTTCCAATGAAGCAAAGTCGA-3'。

the relative expression level of the OsERF940 gene in the homozygous transgenic line was calculated using the expression level of OsERF940 in flower 11 in rice as 1.

As shown in FIG. 2, the expression level of ERF940 in the 8T 2 generation lines is higher than that of the control, which indicates that the obtained 8T 2 generation lines are positive transgenic rice with OsERF940 genes.

Example 2 detection of disease resistance-related Gene expression in OsERF940 Gene-transferred Rice

The T2 generation plants of the rice positively transformed with the OsERF940 gene are used as the following test plants.

Test plants: t of OE3 strain ₂ T of the generation plants, OE5 lines ₂ Generation plants and rice medium flower 11 plants (WT).

Total RNA of leaves (4 weeks seedlings) of the plant to be tested is extracted, reverse transcribed into cDNA, and the expression level of each relevant gene is identified by quantitative PCR using the cDNA as a template. And calculating the relative expression level of the disease-resistant related gene in the transgenic strain by taking the expression level of the disease-resistant related gene in the flower 11 in the rice as 1.

Primers used to identify the OsMYC2 gene were as follows:

an upstream primer: 5'-ATCATGACTAGAGAGGAGCA-3';

a downstream primer: 5'-CCAACACGAGCCTCACAATCA-3';

the primers used to identify the PR1 gene were as follows:

an upstream primer: 5'-GCGCTGCAGGAGGACTACGTA-3';

a downstream primer: 5'-CCCTCCGGCACAAGTATACA-3'.

The primers used to identify the PR4 gene were as follows:

an upstream primer: 5'-CTTCAAGAAGATCGACACAGA-3';

a downstream primer: 5'-CAGAGTGCCAACCTCTTCCA-3'.

The primers used to identify the Chitinase gene were as follows:

an upstream primer: 5'-TTGGCGGCGGTGCAGTGAACA-3';

a downstream primer: 5'-CATGCATATATATCGGTTTCA-3'.

The relative expression level of the corresponding gene in the transgenic line was calculated using the expression level of the gene in flower 11 in rice as 1.

The results are shown in FIG. 3, T ₂ The relative expression level of the disease-resistant related genes in the rice transformed with the OsERF940 gene is higher than that of the rice plant with flower 11 in a wild rice variety, which indicates that the over-expression of the OsERF940 gene can obviously improve the expression of the disease-resistant related genes of the rice such as OsMYC2, PR1, PR4, chitinase and the like.

The result shows that the OsERF940 gene has important regulation and control effects on the expression of the rice disease resistance related gene, and the over-expression of the OsERF940 gene can improve the rice blast resistance by increasing the expression of the disease resistance related gene.

Example 3 Rice blast resistance analysis of transgenic plants

Test plants: t of the rice middle flower 11 plant (WT, control), OE1-OE8 line ₂ And (3) replacing the OsERF940 gene rice plants.

1. Field inspection

The plants to be tested are planted in the field of the test field, and are planted according to the standard requirements of production and inoculation.

FIG. 4A is a photograph showing the occurrence of rice blast of OE3, OE4, OE5 and rice flower 11.

The method for detecting the disease resistance of the field test plants refers to the classification standard of the rice blast resistance of the international paddy rice institute (the national institute of Chinese and south university, shougarmy, 2011.5 months, shuoshi, hunan homonymous and heterologous local Rice resource core germplasm selection).

The results are shown in Table 1 below, and it can be seen that the transgenic lines were all leaf blast resistant compared to the control.

Table 1 shows statistics of disease resistance of the transgenic rice blast strain inoculated

2. Indoor detection:

inoculating filter paper sheet with rice blast bacterial strain (strain GD 08-T13) to starch culture medium (yeast extract 2g, soluble starch 10g, agar powder 15g, adding water to constant volume 1L water), activating and culturing at 26 deg.C for 7 days, inoculating activated mycelium to oat culture medium (sugar-free oatmeal 50 g/broken to powder, sucrose 20g, agar 20g, adding water to constant volume 1L), culturing at 26 deg.C in darkness for 7 days, scraping mycelium after mycelium grows up to the culture medium, and culturing under continuous illumination for 3-4 days to produce spore. Then the rice middle flowers 11 and T of the tested plants in the three-leaf one-heart period are subjected to ₂ Rice with substitution OsERF940 gene OE3 and T ₂ Respectively inoculating rice blast bacteria into the rice OE5 seedlings transformed with the OsERF940 genes: washing spores on the culture medium with sterilized water, and adjusting spore concentration to 5×10 ⁵ And adding Tween20 in two parts per million (W/mL), and uniformly spraying spore liquid on the surface of rice leaves (three leaves and one heart stage rice seedlings) by using a small sprayer (the dosage is that the leaves are all wet).

Immediately covering the seedlings with a custom made plexiglas cover, placing the seedlings in a climatic chamber, dark-treating at 25 ℃ for 24 hours, and then turning into normal light conditions. The onset of disease was investigated 5-7 days after inoculation.

As a result, as shown in FIG. 4B, a photograph of a rice blast laboratory of a different material shows that T is compared with flower 11 (WT) in rice ₂ The occurrence of rice blast of the rice plant transformed with the OsERF940 gene is obviously weaker than that of rice medium flower 11, and the resistance of the rice blast of the transgenic plant is obviously higher than that of a wild type control, which indicates that the overexpression of the OsERF940 gene plays an important role in improving the resistance of the rice blast. The OsERF940 gene can improve the rice blast resistance of the rice by increasing the expression of the rice disease resistance related gene.

SEQUENCE LISTING

<110> institute of biotechnology of national academy of agricultural sciences

Application of <120> OsERF940 protein in improving rice blast resistance

<160> 2

<170> PatentIn version 3.5

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<211> 317

<212> PRT

<213> Oryza sativa L.

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Met Val Pro Ser Ser Arg Lys Val Arg Val Phe Cys Ser Asp Pro Asp

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Pro Val Lys Thr Leu Val Gln Cys Gly Thr Lys Thr Val Lys Asp Ser

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Glu Lys Glu Pro Thr Ser Lys Tyr Arg Gly Val Arg Arg Arg Ala Trp

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Gly Lys Trp Ala Ala Glu Ile Arg Asp Pro Val Arg Lys Ser Arg Lys

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Trp Ile Gly Thr Phe Asn Ser Glu Glu Glu Ala Ala Ala Ala Tyr Leu

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Ala Gln Ser Asn Gln Phe His Glu Glu Leu Met Ala Leu Lys Ile Gln

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Ser Ser Val Ser Glu Gln Glu Asp Leu Ser Ser Ser Val Thr Ile Ser

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Cys Val Ser Ser Ser Gln Ser Cys Asp Gln Lys Ile Gln Ala Lys Pro

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Gln Glu His Lys Arg Val Ser Val Val Val Asn Arg Glu Thr Val Glu

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Gln Lys Phe Lys Ala Gln Pro Gln Ala Gln Lys Ile Lys Ala Gln Pro

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Glu Val Gln Lys Arg Val Ser Val Lys Ile Ser His Glu Thr Glu Asp

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Glu His Leu Leu Asn Leu Pro Ser Thr Pro Lys Gly Lys Glu Ile Ser

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Asp Ala Phe Pro Val Ser Asp Phe Ile Gly Met Ala Asp Val Pro Leu

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Gly Asp Asp Tyr Ile Gly Leu Ala Asp Ile Ser His Leu Pro Leu Pro

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<210> 2

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<213> Oryza sativa L.

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atggtgccct cttcacggaa agtccgtgtt ttctgctctg atcctgatgc cactgactcc 60

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atggaaaact ccaaggcatc caaacctgtt aagacccttg ttcaatgtgg cacaaagact 180

gtaaaggatt ccgagaagga accgacgagc aaatacaggg gtgtgcgccg gcgggcgtgg 240

ggcaaatggg ctgcagaaat acgtgaccct gtgcgaaaat caaggaaatg gattggcaca 300

tttaacagtg aggaggaggc cgctgcggca tatcttgcac aatcaaacca gttccatgaa 360

gagttgatgg ccttgaaaat ccaatcttct gtatcagagc aggaagattt gtcaagctct 420

gttactatat cctgcgtgtc ctcatctcag tcatgtgacc agaagattca ggcaaagcca 480

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gcacagccac aagctcagaa gattaaggca cagccagaag tgcagaagag agtgtcagtg 600

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aaagagattt cgatgggcgc tgttcttggc aggatagatg agattcctgt gagcaactgt 720

gtgggccata ttgatgaatt tccacctgat gatttcacta ggcttgcaga tgcgttccca 780

gttagtgatt ttatcggcat ggcagatgtg ccactgggcg atgattacat tggccttgca 840

gacatcagcc acctgccgct gcctatcaca gatctgaagt tcgacctcaa tgcagaactc 900

aactgggacg ggttcgactt tgcttcattg gaacaagaac tcaactgtct ttga 954

Claims

1. The use of any of the following 1) -3) for increasing the resistance of plants to rice blast;

1) Protein OsERF940;

2) A nucleic acid molecule encoding the protein OsERF940;

3) A recombinant vector, expression cassette or recombinant bacterium comprising a nucleic acid molecule encoding a protein OsERF940;

the protein OsERF940 is as follows (1) or (2):

(1) A protein consisting of an amino acid sequence shown as a sequence 1 in a sequence table;

(2) A protein formed by adding a tag sequence at the tail end of an amino acid sequence shown in a sequence 1 in a sequence table;

the plant is rice.

2. The use according to claim 1, characterized in that:

the nucleic acid molecule encoding the protein OsERF940 is a DNA molecule of any one of the following 1) to 2):

1) The coding region is a DNA molecule shown as a sequence 2 in a sequence table;

2) DNA molecules shown in the 1 st-951 st positions of the sequence 2 in the sequence table.

3. Use of any one of the substances 1) to 3) according to claim 1 or 2 for cultivating rice against rice blast.

4. Use of any one of the substances 1) -3) according to claim 1 or 2 for cultivating rice with increased resistance to rice blast.

5. A method for breeding transgenic plants with increased resistance to rice blast, which is 1) or 2):

1) The method comprises the following steps: increasing the content and/or activity of the protein OsERF940 in the target plant to obtain a transgenic plant;

2) The method comprises the following steps: improving the expression of a nucleic acid molecule encoding a protein OsERF940 in a target plant to obtain a transgenic plant;

the transgenic plant has a rice blast resistance higher than that of the plant of interest;

the protein OsERF940 is as follows (1) or (2):

the plant is rice.

6. The method according to claim 5, wherein:

the method for improving the content and/or activity of the protein OsERF940 in the target plant or improving the expression of the nucleic acid molecule encoding the protein OsERF940 in the target plant is to introduce the nucleic acid molecule encoding the protein OsERF940 into the target plant.