Antibacterial peptide SAFM470 and application thereof in prevention and treatment of rice blast

Abstract

The present application relates to the field of biotechnology, in particular to an antibacterial peptide SAFM470 and its application in preventing and treating rice blast, and discloses the antibacterial peptide SAFM470 and its application in preventing and treating rice blast. The present application finds that the antibacterial peptide SAFM470 can be used as a medicament to prevent and treat rice blast. The antibacterial peptide SAFM470 has obvious inhibitory effect on appressorium of Magnaporthe oryzae, when the concentration of the antibacterial peptide SAFM470 is 0.14 mu g / mu L, the appressorium formation rate of spores of Magnaporthe oryzae is 1.2%; and the antibacterial peptide SAFM470 can significantly inhibit the infection of Magnaporthe oryzae on rice leaves. Therefore, the antibacterial peptide SAFM470 can be used to improve the resistance of rice to Magnaporthe oryzae.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to an antimicrobial peptide SAFM470 and its application in the prevention and control of rice blast. Background Technology

[0002] Rice blast is one of the four major diseases of rice. Caused by the rice blast fungus (Magnaporthe oryzae), rice blast is characterized by its rapid spread and high destructive potential. In severe cases, it can lead to plant death and complete crop failure, posing a significant threat to agricultural production. It occurs in all rice-growing regions worldwide, with leaf and node involvement being the most common. Infections can cause varying degrees of yield reduction, especially early and severe neck blast or node blast, which can result in whiteheads and even total crop failure.

[0003] Rice blast fungus infects rice through a direct breach of the host epidermis by the mechanical force generated by appressoriums. This is the most characteristic pathogenic mechanism of the fungus and the reason for its immense destructive power. The principle of controlling rice blast is to prevent the germination and invasion of fungal spores, or to inhibit or kill the already invading fungal hyphae, either externally or internally, thereby achieving preventative and curative effects. Currently, the most common, efficient, and rapid method for controlling rice blast is pesticide spraying. However, pesticide spraying has certain limitations; it not only pollutes the environment but also increases the fungus's resistance to pesticides. In agricultural production, antimicrobial peptides (AMPs) have significant potential and unique advantages in controlling rice blast. Many antimicrobial peptides have strong inhibitory or killing effects on multiple growth stages of the fungus, including mycelial growth, spore germination, and appressorium formation. Antimicrobial peptides are essentially proteins or polypeptides and do not cause residues or environmental pollution like some chemical pesticides. They have low toxicity to non-target organisms (such as humans, livestock and beneficial insects), which meets the requirements of agricultural product safety and the development of ecological agriculture.

[0004] Currently, most disease control measures rely on chemical pesticides. However, chemical control is not only costly and pollutes the environment, but its effectiveness is also unsatisfactory; at the same time, food safety is seriously affected. With societal progress, people are increasingly aware of the harm that long-term, large-scale use of chemical pesticides poses to the ecological environment and human health. Biological control can effectively overcome these drawbacks, and therefore, biological pesticides are receiving increasing attention.

[0005] Antimicrobial peptides are a class of natural small-molecule proteins with antimicrobial activity, widely distributed in the biological world. They are an important component of the body's nonspecific immunity and exhibit antimicrobial activity both in vivo and in vitro. Antimicrobial peptides possess broad biological activity, exhibiting strong inhibitory effects against bacteria, fungi, protozoa, viruses, and tumors. Their advantages, such as high efficiency, environmental friendliness, and non-induction of drug resistance in pathogens, make them promising candidates for biological control and medicine. For example, Chinese invention application CN118791582A discloses an antimicrobial peptide, OsSspr1b, for the control of rice blast disease, which shows significant inhibitory effects on the appressorium of rice blast fungus.

[0006] However, little is known about new agents for the effective control of rice blast, and the development of new antimicrobial peptides for the prevention and control of rice blast is of practical significance. Summary of the Invention

[0007] The purpose of this invention is to provide an antimicrobial peptide, SAFM470, and its application in the control of rice blast. This invention reveals the regulatory mechanism of the rice antimicrobial peptide SAFM470. Based on its functions of inhibiting the germination of rice blast fungus, inhibiting appressorium formation, and inhibiting the infection of rice leaves by rice blast fungus, it is effectively applied to the control of rice blast, and highly efficient agents are developed to achieve effective control of rice blast.

[0008] To achieve the above objectives, the present invention provides the following solution:

[0009] The present invention provides an antimicrobial peptide SAFM470, the amino acid sequence of which is shown in SEQ ID No. 4.

[0010] The present invention provides a gene encoding the above-mentioned antimicrobial peptide SAFM470, the nucleotide sequence of which is shown in SEQ ID No.3.

[0011] This invention provides a biological material containing the above-mentioned coding gene.

[0012] Optionally, the biomaterial includes a recombinant vector and recombinant bacteria.

[0013] This invention provides the application of the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, or the above-mentioned biological material in the preparation of rice blast-resistant products.

[0014] The present invention provides a product for resisting rice blast, the product comprising the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, or the above-mentioned biological material.

[0015] This invention provides the application of the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, the above-mentioned biological material, or the above-mentioned product in improving the resistance of rice to rice blast fungus.

[0016] The present invention provides a method for improving the resistance of rice to rice blast fungus, comprising the steps of treating rice with the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, the above-mentioned biological material or the above-mentioned product.

[0017] This invention provides the application of the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, the above-mentioned biological material, or the above-mentioned product in the prevention and control of rice blast fungus infection.

[0018] The present invention provides a method for preventing and controlling rice blast fungus infection, comprising the steps of treating rice with the above-mentioned antimicrobial peptide SAFM470, the above-mentioned encoding gene, the above-mentioned biological material or the above-mentioned product.

[0019] The present invention discloses the following technical effects:

[0020] This invention has discovered that the antimicrobial peptide SAFM470 can be used as a pesticide to control rice blast. SAFM470 exhibits a significant inhibitory effect on the appressorium of rice blast fungus; when the concentration of SAFM470 is 0.14 μg / μL, the appressorium formation rate of rice blast fungus spores is 1.2%; moreover, it can significantly inhibit the infection of rice leaves by rice blast fungus. Therefore, the antimicrobial peptide SAFM470 of this invention can be used to improve the resistance of rice to rice blast fungus. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 Image A shows the purification process of the antimicrobial peptide SAFM470 protein. Image B shows the Western blot assay of the purified protein, with lanes 1-6 representing PET-supernatant (empty plasmid control group supernatant), SAFM470-supernatant (supernatant), Elution 1, Elution 2, Elution 3, and Elution 4, respectively.

[0023] Figure 2 The graph shows the inhibition of appressorium formation by the antimicrobial peptide SAFM470 (purified rice SAFM470-HIS protein). In the graph, A is the appressorium formation of rice blast fungus; B is the statistical result of appressorium formation.

[0024] Figure 3 The images show the results of in vitro inoculation of rice leaves with the antimicrobial peptide SAFM470 (rice SAFM470-HIS supernatant) inhibiting the infection of rice blast fungus. Among them, A is the result of in vitro inoculation of rice leaves with rice blast fungus; B is the statistical result of the length of rice leaf lesions.

[0025] Figure 4 Figure showing the results of spray inoculation of rice leaves to inhibit rice blast fungus infection by the antimicrobial peptide SAFM470 (rice SAFM470-HIS supernatant). Detailed Implementation

[0026] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0027] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0028] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0029] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0030] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0031] The amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No. 1, specifically:

[0032] MVSLRLPLILLSLLAISFSCSAAPPPVYDTEGHELSADGSYYVLPASPGHGGGLTMAPRVLPCPLLVAQETDERRKGFPVRFTPWGGAAAPEDRTIRVSTDVRIRFNAATICVQSTEWHVGDEPLTGARRVVTGPLIGPSPSGRENAFRVEKYGGGYKLVSCRDSCQDLGVSRDGARAWLGASQPPHVVVFKKARPSPPE.

[0033] The complete nucleotide sequence of the antimicrobial peptide SAFM470 gene is shown in SEQ ID No. 2, which also includes the sequence encoding the signal peptide, specifically:

[0034] ATGGTTAGCCTCCGCCTCCCCCTCATACTCCTCTCCCTCCTGGCCATCTCCTTCTCATGCAGCGCCGCGCCGCCGCCGGTGTACGACACGGAGGGCCACGAGCTGAGCGCCGACGGGAGCTACTACGTCCTCCCGGCTAGCCCCGGCCACGGAGGGGGCCTCACGATGGCGCCCCGCGTGCTCCCCTGCCCGCTCCTCGTGGCGCAGGAGACGGACGAGCGCCGCAAGGGGTTCCCCGTGCGCTTCACCCCGTGGGGCGGCGCCGCGGCGCCGGAGGACAGGACCATCCGCGTCTCGACCGACGTCCGCATCCGCTTCAACGCCGCGACGATCTGCGTGCAGTCCACCGAGTGGCATGTCGGCGACGAGCCGCTCACGGGGGCGCGGCGCGTGGTGACGGGGCCGTTGATCGGGCCGAGCCCGAGCGGGCGGGAGAACGCGTTCCGCGTGGAGAAGTACGGCGGTGGGTACAAGCTGGTGTCGTGCAGGGACTCGTGCCAGGACCTGGGCGTGTCAAGGGACGGCGCGCGGGCGTGGCTGGGCGCGAGCCAGCCGCCTCACGTCGTGGTCTTCAAGAAGGCCAGGCCAAGCCCACCAGAGTAA。

[0035] The nucleotide sequence of the antimicrobial peptide SAFM470 gene after removing the signal peptide coding sequence is shown in SEQ ID No. 3, specifically:

[0036] .

[0037] The amino acid sequence of the antimicrobial peptide SAFM470 after removing the signal peptide coding sequence is shown in SEQ ID No. 4, specifically:

[0038] MAPPPVYDTEGHELSADGSYYVLPASPGHGGGLTMAPRVLPCPLLVAQETDERRKGFPVRFTPWGGAAAPEDRTIRVSTDVRIRFNAATICVQSTEWHVGDEPLTGARRVVTGPLIGPSPSGRENAFRVEKYGGGYKLVSCRDSCQDLGVSRDGARAWLGASQPPHVVVFKKARPSPPE.

[0039] Example 1: Obtaining the antimicrobial peptide SAFM470 (amino acid sequence as shown in SEQ ID No. 4)

[0040] 1. Based on the CDS sequence of gene SAFM470 (as shown in SEQ ID No. 3), primers SAFM470-F and SAFM470-R were designed. The primer sequences are as follows:

[0041] SAFM470-F: cagcaaatgggtcgcggatccATGGCGCCGCCGCCGGTG (SEQ ID No. 5);

[0042] SAFM470-R: ctcgagtgcggccgcaagcttCTCTGGTGGGCTTGGCCT (SEQ ID No. 6).

[0043] 2. Construction of the SAFM470-HIS vector: Using the genome of rice variety Zhonghua 11 as a template, the CDS sequence of the SAFM470 gene was amplified using primers SAFM470-F and SAFM470-R. The amplified fragment was ligated into the PET-28a HIS vector (Thermo Fisher Scientific, catalog number 10265443) after digestion with BamHI and HindIII using seamless cloning technology. The ligated plasmid was transformed into *E. coli* using a heat shock method. Positive clones were selected for testing, and the SAFM470-HIS vector with the correct sequence was finally obtained.

[0044] 3. Induction of SAFM470-HIS protein expression (SAFM470 protein with HIS tag sequence fused to its C-terminus): After the SAFM470-HIS vector was correctly sequenced, the plasmid was transformed into Escherichia coli strain BL21(DE3) by heat shock. Positive clones were selected and cultured at 37℃ until the OD value reached 0.6, and then induced at 28℃ for 10 h with an IPTG concentration of 1 mM.

[0045] 4. SAFM470-HIS protein purification:

[0046] (1) Centrifuge 150 mL of bacterial cells at 10,000 rpm for 2 min and remove the supernatant; add 15 mL of HIS binding / washing buffer (20 mM Na2HPO4, 0.5 M NaCl, 35 mM imidazole, pH adjusted to 7.4) to the precipitated bacterial cells to suspend the cells, and then add 300 µL of 10 mg / mL lysozyme and 150 µL of 0.1 M benzyl sulfonyl fluoride (PMSF); and slowly shake at 4℃ for 30 min to digest the enzymes.

[0047] (2) After the enzymatic hydrolysis process is completed, the bacterial cells are broken by ultrasonication and then centrifuged at 4℃ for 10 min at 10000 rpm.

[0048] (3) Take 200 µL of supernatant and label it as Input (i.e., SAFM470-supernatant (supernatant)). Transfer the remaining supernatant to a pre-equilibrated HIS protein resin flow column, adjust the flow rate to 7-9 s / drop, and collect 200 µL of effluent and label it as Flowthroµgh. Add 15 mL of water to the bacterial residue and suspend it. Take 200 µL and label it as Bacteriapellet (bacterial cell). The entire process is carried out in a 4℃ refrigerator to prevent protein denaturation.

[0049] (4) Add 5 mL binding / washing buffer, control the flow rate at 4-6 s / drop, collect 200 µL of effluent at the bottom of the column and label it Wash1; repeat this step once and collect 200 µL of effluent and label it Wash2.

[0050] (5) Elute the target protein with 1 mL of HIS elution buffer (20 mM Na2HPO4, 0.5 M NaCl, 500 mM imidazole, pH adjusted to 7.4), and collect the eluent (containing the target protein) and label it Elution 1. Repeat this step 3 times and label it Elution 2, Elution 3 and Elution 4 respectively.

[0051] 5. Preparation of PET-supernatant (empty plasmid control group supernatant):

[0052] Escherichia coli strain BL21(DE3) was transformed by heat shock. Positive clones were selected and cultured at 37°C until the OD value reached 0.6. They were then induced at 28°C for 10 h with an IPTG concentration of 1 mM. Steps (1) and (2) were then performed to obtain PET-supernatant (empty plasmid control group supernatant).

[0053] 6. Results:

[0054] The purified protein (sequence shown in SEQ ID No. 4, i.e., the antimicrobial peptide SAFM470 described in this application) was detected by Coomassie Brilliant Blue staining and Western blot assay, and then the concentration of the purified protein was determined. The Coomassie Brilliant Blue staining results are shown below. Figure 1 As shown in A, the results of Western blot analysis of protein expression are as follows: Figure 1 As shown in B in the diagram.

[0055] Example 2: SAFM470-HIS purified protein inhibits rice blast fungus appressorium formation.

[0056] Wild-type rice blast fungus strain B157 (disclosed in the literature "MoWhi2regulates appressorium formation and pathogenicity via the MoTor signalling pathway in Magnaporthe oryzae") was activated on OA medium and cultured in the dark at 25°C for 3 days, followed by light culture for 4 days. Sterile ddH2O was added to the OA culture dish, and the mycelia were gently scraped off with an inoculation loop to elute the rice blast fungus spores from the medium. The eluent was filtered through a filter cloth to obtain a spore suspension. The spore suspension was placed in a 2 mL centrifuge tube and centrifuged at 10,000 rpm for 1 min. The supernatant was discarded (avoiding the bottom spores from being poured out). Sterile ddH2O was added, and the spore concentration was adjusted to no less than 3 × 10⁻⁶ using a hemocytometer. 4 The concentration of SAFM470-HIS purified protein in the suspension was set up as follows: 0 (i.e., using only buffer solution, buffer solution: 20 mM Na2HPO4 and 0.5 M NaCl, pH adjusted to 7.4), 0.06, 0.08, 0.1, 0.12, and 0.14 µg / µL. The suspension was dropped onto a hydrophobic glass slide, and the formation of rice blast fungus spores and appressoria was observed under a microscope after 24 h.

[0057] like Figure 2 As shown, SAFM470-HIS protein has a significant inhibitory effect on rice blast fungus appressorium. The appressorium formation rate of rice blast fungus spores in H2O was 97.79%. When the concentration of SAFM470-HIS was 0.14 µg / µL, the appressorium formation rate of rice blast fungus spores was 1.2%, which was 96.59% lower than that of the control group, showing a highly significant difference. This result indicates that the antimicrobial peptide SAFM470 has the potential to be used as an agent for the prevention and control of rice blast.

[0058] Example 3: SAFM470-HIS supernatant protein inhibits rice blast fungus infection in rice leaves.

[0059] 1. Preparation of water agar:

[0060] Prepare a 3% water agar solution with water, sterilize at 110℃ for 30 minutes, and after the water agar solution has cooled slightly, add the agonist (Kinetin) to make its working concentration 2µg / mL, 100µg / mL carbenicillin, and 100µg / mL streptomycin. Mix well and pour the liquid into a 15 cm culture dish.

[0061] 2. In vitro inoculation:

[0062] (1) Take 150 mL of the bacterial cells obtained in step “3. Inducing the expression of SAFM470-HIS protein (SAFM470 protein has a HIS tag sequence fused to its C-terminus)” in Example 1, centrifuge at 10000 rpm for 2 min, and remove the supernatant; add 15 mL of HIS suspension buffer (20 mM Na2HPO4, 0.5 M NaCl, pH adjusted to 7.4) to the precipitated bacterial block to suspend the bacterial cells, break the bacterial cells by sonication, and then centrifuge at 4℃ for 10 min at 10000 rpm, and remove the supernatant to obtain SAFM470-HIS supernatant protein.

[0063] (2) Wild rice blast fungus strain B157 was activated on OA medium under the same culture conditions and spore suspension preparation process as in Example 2. The spore concentration was adjusted to 1×10⁻⁶ using a hemocytometer. 6 0.1% Gelatin (final Gelatin concentration approximately 0.01%) was added to the obtained spore solution at a concentration of 0.85 μg / μL. SAFM470-HIS protein supernatant or empty vector (i.e., PET-supernatant from step "5. Preparation of PET-supernatant (empty plasmid control group supernatant)" in Example 2) was also added to achieve a suspension concentration of 0.85 μg / μL, resulting in a spore mixture used for inoculation with rice blast fungus. These two treatments were designated as the 0.85 μg / μL group and the empty vector group, respectively. Equal volumes of a control group (water) and a buffer solution group (20 mM Na2HPO4 and 0.5 M NaCl, pH 7.4) were also included.

[0064] (3) Take CO39 rice blast disease-susceptible material (provided by China National Rice Research Institute, available to the public) at the 4-leaf stage. Take about 5cm of the lower part of the second leaf from the bottom. Make two wounds on the main vein with a needle without penetrating it. Gently wipe the wounded leaf with alcohol 3 times, and then wash it with ddH2O 3 times. Place the cleaned leaf face up on a water agar medium and press the two ends of the leaf with a water agar block to prevent the leaf from curling. Use a pipette to drop 10µL of spore mixture onto the wound. After standing for 30s, put the inoculated CO39 leaf into a 25℃ constant temperature incubator and culture in the dark for 48h. Then restore light and culture under light and dark conditions for 7d. Observe the disease situation.

[0065] 3. Results:

[0066] Vaccination status as follows Figure 3 As shown, the results indicate that the addition of 0.85 μg / μL of SAFM470-HIS protein supernatant can reduce the lesion area compared with the control group, indicating that the antimicrobial peptide SAFM470 can inhibit the infection of rice leaves by rice blast fungus.

[0067] Example 4: SAFM470-HIS supernatant protein inhibits rice blast fungus infection in rice leaves.

[0068] 1. Obtaining seedlings:

[0069] Seeds of rice variety CO39 were soaked at 37℃ for 1 day and kept moist at 37℃ for 2 days to promote germination. Most of the germinations were about 0.5cm long. Select germinating seeds with uniform growth and sow 5 seeds per pot in each pot. Keep the soil appropriately dry after sowing. When the rice has grown about 2 leaves, retain 3 CO39 seedlings with uniform growth. When the CO39 seedlings grow to the 4-leaf stage or tillering stage, they can be used for inoculation experiments.

[0070] 2. Rice spraying inoculation:

[0071] (1) Take 150 mL of bacterial cells obtained in step 3 of Example 1, “Inducing the expression of SAFM470-HIS protein (SAFM470 protein has a HIS tag sequence fused to its C-terminus),” centrifuge at 10000 rpm for 2 min, and remove the supernatant; add 15 mL of HIS suspension buffer (20 mM Na2HPO4, 0.5 M NaCl, pH adjusted to 7.4) to the precipitated bacterial block to suspend the bacterial cells, break the bacterial cells by sonication, and then centrifuge at 4℃ for 10 min at 10000 rpm, and remove the supernatant to obtain SAFM470-HIS supernatant protein.

[0072] (2) Wild rice blast fungus strain B157 was activated on OA medium under the same culture conditions and spore suspension preparation process as in Example 2. The spore concentration was adjusted to 1×10⁻⁶ using a hemocytometer. 6 0.1% Gelatin (final Gelatin concentration approximately 0.01%) was added to the obtained spore solution at a concentration of 1 / 10 volume. SAFM470-HIS protein supernatant or empty vector (i.e., PET-supernatant from step "5. Preparation of PET-supernatant (empty plasmid control group supernatant)" in Example 2) was added to achieve a suspension concentration of 0.85 μg / μL, resulting in a spore mixture used for inoculation with rice blast fungus. These two treatments were designated as the SAFM470 group and the empty vector group, respectively. An additional group was prepared by adding equal volumes of ck (water) and buffer (20 mM Na2HPO4 and 0.5 M NaCl, pH 7.4).

[0073] (3) Prepare rice CO39 at the 4-leaf stage to tillering stage. Seal the rice to be inoculated in a roll made of transparent PVC film. Inoculate in the evening. Spray the spore mixture with added gelatin onto the rice leaves using a small spray bottle until the leaves are thoroughly wet. Cover the top with a damp plastic wrap to keep it moist. Incubate at 22℃ in the dark for 1 day, followed by alternating light and dark periods of 16 hours and 8 hours. Observe daily. When obvious lesions appear on the leaves, take photos of the leaves. Alternatively, observe and take photos when the diseased leaves show the greatest difference in disease incidence. Attach the back of the diseased leaves to an A4 sheet with double-sided tape.

[0074] 3. Results:

[0075] Vaccination status as follows Figure 4 As shown, the results indicate that the addition of 0.85 μg / μL of SAFM470-HIS protein supernatant can reduce the lesion area compared with the control group, indicating that the antimicrobial peptide SAFM470 can inhibit the infection of rice leaves by rice blast fungus.

[0076] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A gene for the antimicrobial peptide SAFM470, characterized in that, The nucleotide sequence of the antimicrobial peptide SAFM470 gene is shown in SEQ ID No.

3.

2. A biomaterial containing the antimicrobial peptide SAFM470 gene as described in claim 1.

3. The biomaterial according to claim 2, characterized in that, The biomaterials include recombinant vectors and recombinant bacteria.

4. The application of the antimicrobial peptide SAFM470 gene of claim 1, the antimicrobial peptide SAFM470 encoded by the antimicrobial peptide SAFM470 gene of claim 1, or the biomaterial of claim 2 or 3 in the preparation of rice blast-resistant products, characterized in that, The amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No.

4.

5. A product resistant to rice blast, characterized in that, The product includes the antimicrobial peptide SAFM470 gene as described in claim 1 or the biomaterial as described in claim 2 or 3.

6. The application of the antimicrobial peptide SAFM470 gene of claim 1, the antimicrobial peptide SAFM470 encoded by the antimicrobial peptide SAFM470 gene of claim 1, the biomaterial of claim 2 or 3, or the product of claim 5 in improving the resistance of rice to rice blast fungus, characterized in that, The amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No.

4.

7. A method for improving the resistance of rice to rice blast fungus, characterized in that, The method includes the steps of treating rice with the antimicrobial peptide SAFM470 gene as described in claim 1, the antimicrobial peptide SAFM470 encoded by the antimicrobial peptide SAFM470 gene as described in claim 1, the biomaterial as described in claim 2 or 3, or the product as described in claim 5; the amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No.

4.

8. The application of the antimicrobial peptide SAFM470 gene of claim 1, the antimicrobial peptide SAFM470 encoded by the antimicrobial peptide SAFM470 gene of claim 1, the biomaterial of claim 2 or 3, or the product of claim 5 in the prevention and control of rice blast fungus infection, characterized in that, The amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No.

4.

9. A method for preventing and controlling rice blast fungus infection, characterized in that, The method includes the steps of treating rice with the antimicrobial peptide SAFM470 gene as described in claim 1, the antimicrobial peptide SAFM470 encoded by the antimicrobial peptide SAFM470 gene as described in claim 1, the biomaterial as described in claim 2 or 3, or the product as described in claim 5; the amino acid sequence of the antimicrobial peptide SAFM470 is shown in SEQ ID No. 4.

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