KR102633074B1 - Composition for controlling plant diseases using Aspergillus montenegroi SFC20200425-M27 strain and method for manufacturing the same - Google Patents

Abstract

The present invention provides a composition for controlling plant diseases comprising Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain, a culture medium of the strain, a culture supernatant of the strain, an extract of the culture supernatant, or a mixture thereof as an active ingredient, and It relates to a method for controlling plant diseases using this, and the composition for controlling plant diseases of the present invention exhibits excellent plant disease control effects against various plant diseases.

Description

Composition for controlling plant diseases using Aspergillus montenegroi SFC20200425-M27 strain and method for manufacturing the same}

The present invention relates to a composition for controlling plant diseases using Aspergillus montenegroi SFC20200425-M27 strain, a method for producing the same, and a method for controlling plant diseases using the same.

As the world population is expected to increase rapidly and reach 9.7 billion in 2050, there is a need to increase food production by at least 50% to 110%. However, in order to increase food production in limited farmland, efforts are needed to minimize crop yield loss due to plant diseases. Pesticides are the most effective means of controlling plant diseases, but excessive use of chemical pesticides causes problems such as drug resistance, human toxicity, and environmental pollution. Therefore, the development of new eco-friendly plant disease control methods to reduce or replace the use of chemical pesticides is urgently needed. For this purpose, natural materials such as plant extracts, microorganisms, or secondary metabolites of microorganisms are being actively developed as new plant disease control agents as substitutes for chemical pesticides.

Of the more than 23,000 types of microbial secondary products known to date, about 42% are produced by fungi. Since the discovery of penicillin in the fungus of the Penicillium genus, the fungus has received much attention over the past few years due to its medical value and utility in various fields, and various antibacterial substances have been isolated from the fungus. In particular, strobilurin, produced by wood-rotting basidiomycetes, has a unique mechanism of action that inhibits mitochondrial respiration of plant pathogens, and various derivative compounds have been synthesized, currently occupying more than 23% of the global fungicide market. Griseofulvin, produced by the ascomycete Penicillium griseofulvum , has received attention as an active plant disease control agent that inhibits the normal spore germination of the onion gray mold fungus ( Botrytis allii ). Pestalachloride A-C produced by Pestalotiopsis adusta has antibacterial activity against plant pathogens of the genus Fusarium and Verticillium , and Stagonospora nodorum . ) produces (+)-4'methoxy-(2 S )-methylbutyrophenone, alternariol, (3 R )-(―)-mellein methyl ester ) shows antibacterial activity against Fusarium solani, and (3 R , 4 R )-(―)-4-hydroxymellein produced by Sphaeropsis sapinea . It exhibits antibacterial activity against various plant pathogens, such as pepper anthracnose ( Colletotrichum acutatum ), onion white silk fungus ( Sclerotium rolfsii ), and tomato gray mold fungus ( Botrytis cinerea ).

Aspergillus montenegroi was first isolated from soil samples in Japan in 1996 and was initially named Emericella montenegroi . It was discovered as a marine microorganism in the waters off Korea in 2016. However, until now, nothing is known about the chemical properties or biological activities of secondary metabolites produced by Aspergillus montenegroi .

Because fungi produce various types of secondary metabolites with antibacterial activity, they can be actively used to develop new crop protection agents.

(Non-patent Document 1) Mycobiology 2016 December, 44(4): 237-247 (Non-patent Document 2) J Pharmacol Sci 116, 283-295 (2011)

In order to develop an eco-friendly plant disease control agent, the present inventor was investigating the control effect of various strains isolated from the ocean against various plant diseases. Aspergillus montenegroi ( Aspergillus montenegroi ) KACC 83033BP (SFC20200425-M27), its culture medium, and its It was discovered that the culture filtrate or the organic solvent extract of the culture filtrate had an excellent control effect on various plant diseases.

The purpose of the present invention is to provide Aspergillus Montenegroi SFC20200425-M27 strain deposited with accession number KACC 83033BP.

Another object of the present invention is to control plant diseases comprising the Aspergillus Montenegroi SFC20200425-M27 (accession number: KACC 83033BP) strain, the culture fluid of the strain, the culture supernatant of the strain, the extract of the culture supernatant, or a mixture thereof as an active ingredient. To provide a composition for use.

Another object of the present invention is to provide a method for producing the composition for controlling plant diseases.

Additionally, the present invention provides a method for controlling plant diseases using the composition for controlling plant diseases.

The present invention provides Aspergillus Montenegroi SFC20200425-M27 strain deposited with accession number KACC 83033BP.

The composition for controlling plant diseases according to the present invention contains Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain, a culture fluid of the strain, a culture supernatant of the strain, an extract of the culture supernatant, or a mixture thereof as an active ingredient. do.

In one embodiment of the present invention, the plant disease may be selected from rice blast, tomato blight, wheat red rust, barley powdery mildew, and pepper anthracnose.

In one embodiment, the composition for controlling plant diseases may further include one or two or more selected from carriers, surfactants, and auxiliaries.

A method of manufacturing a composition for controlling plant diseases according to an embodiment may include forming a culture solution by culturing the Aspergillus Montenegroi SFC20200425-M27 (Accession Number: KACC 83033BP) strain.

In addition, the method of producing the composition for controlling plant diseases may further include the step of removing strains from the culture medium to prepare a culture supernatant, and preparing an extract of the culture supernatant by treating the culture supernatant with an extraction solvent. More may be included. The extraction solvent may be ethyl acetate or butanol.

Additionally, the present invention provides a method for controlling plant diseases using the composition for controlling plant diseases. The plant disease control method may include direct or indirect spraying, volatilization, or seed treatment.

The composition for controlling plant diseases according to the present invention contains Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain, a culture fluid of the strain, a culture supernatant of the strain, an extract of the culture supernatant, or a mixture thereof as an active ingredient. Therefore, it can show a high control effect against various plant diseases such as rice blast, tomato blight, wheat red rust, barley powdery mildew, and pepper anthracnose.

In addition, the method for producing a composition for controlling plant diseases according to an embodiment of the present invention is simple and easy to use industrially under mild conditions.

Furthermore, the culture supernatant extract obtained by extracting the culture supernatant according to an embodiment of the present invention with a specific solvent can exhibit a superior plant disease control effect.

The composition for controlling plant diseases according to an embodiment of the present invention is easily and effectively selected from rice blast fungi, tomato blight fungi, wheat red rust fungi, barley powdery mildew, and pepper anthracnose fungi by using the plant disease control method according to the present invention. It can control one or more plant pathogens.

Figure 1 is a photograph showing the flora of Aspergillus montenegroi SFC20200425-M27 strain cultured on PDA (potato dextrose agar) medium.
Figure 2 is a diagram showing a phylogenetic tree created based on the Camodulin gene (CaM) sequence analysis of the Aspergillus montenegroi SFC20200425-M27 strain.
Figure 3 is a diagram showing the process of obtaining a culture medium of the Aspergillus montenegroi SFC20200425-M27 strain, a culture supernatant of the strain, and an extract of the culture supernatant.

Hereinafter, the composition for controlling plant diseases using the Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain of the present invention will be described in detail. At this time, if there is no other definition in the technical and scientific terms used, they have meanings commonly understood by those skilled in the art to which this invention pertains, and the following description will not unnecessarily obscure the gist of the present invention. Descriptions of possible notification functions and configurations are omitted.

As used herein, the singular forms “a,” “an,” and “the” are intended to also include the plural forms, unless the context clearly dictates otherwise.

In addition, the numerical range used in the present invention includes the lower limit and upper limit and all values within the range, increments logically derived from the shape and width of the defined range, all double-defined values, and the upper limit of the numerical range defined in different forms. and all possible combinations of the lower bounds. Unless otherwise specified in the specification of the present invention, values outside the numerical range that may occur due to experimental error or rounding of values are also included in the defined numerical range.

As used in the present invention, “comprises” is an open description with the same meaning as expressions such as “comprises,” “contains,” “has,” or “characterized by” elements that are not additionally listed; Does not exclude materials or processes.

“Control” in the present invention includes the meaning of preventing, avoiding, removing or killing diseases or pests.

The present invention provides Aspergillus Montenegroi SFC20200425-M27 strain deposited with accession number KACC 83033BP.

The strain may have the calmodulin gene (CaM) base sequence of SEQ ID NO: 1.

The strain may have been isolated from mudflat soil, and specifically, it may have been isolated from soil collected from the Suncheon Bay mudflat (Haksin-ri, Byeolryang-myeon, Suncheon-si, Jeollanam-do).

The strain may have plant disease control activity.

The plant diseases include rice blast (pathogen: Magnaporthe oryzae ), tomato blight (pathogen: Phytophthora infestans ), wheat red rust (pathogen: Puccinia triticina ), barley powdery mildew (pathogen: Blumeria graminis f. sp. hordei ), and pepper anthracnose (pathogen). : Colletotrichum coccodes ), etc., but is not limited to this.

In addition, the present invention provides a method for controlling plant diseases comprising Aspergillus Montenegroi SFC20200425-M27 (accession number: KACC 83033BP) strain, a culture fluid of the strain, a culture supernatant of the strain, an extract of the culture supernatant, or a mixture thereof as an active ingredient. A composition is provided.

In one embodiment, the culture medium of the strain may be obtained by culturing the strain, and the culture supernatant may be obtained by centrifuging the culture medium in which the strain was cultured or filtering it with gauze and filter paper, that is, physically filtering the culture medium. The extract of the supernatant may be obtained by extracting the culture supernatant with ethyl acetate or butanol.

In one embodiment, the plant disease is rice blast (pathogen: Magnaporthe oryzae ), tomato blight (pathogen: Phytophthora infestans ), wheat red rust (pathogen: Puccinia triticina ), barley powdery mildew (pathogen: Blumeria graminis f. sp. hordei ), pepper anthracnose (pathogen: Colletotrichum coccodes ), etc., but is not limited to this.

In one embodiment, the plant may be any one or more selected from the group consisting of rice, tomato, barley, wheat, and pepper.

According to one embodiment, the composition for controlling plant diseases is rice blast (pathogen: Magnaporthe oryzae ), tomato late blight (causing bacteria: Phytophthora infestans ), wheat red rust (causing bacteria: Puccinia triticina ), and pepper anthracnose (causing bacteria: Colletotrichum coccodes ). It can show a better control effect against selected plant diseases.

The composition for controlling plant diseases according to one embodiment may further include one or two or more selected from carriers, surfactants, and auxiliaries.

The carrier may be a liquid carrier or a solid carrier. The liquid carrier may be one or two or more selected from alcohols, ketones, aromatic hydrocarbons, aliphatic hydrocarbons, esters, nitriles, ethers, amides, halogenated hydrocarbons, dimethyl sulfoxide, vegetable oils, and water, but is not limited thereto.

Specifically, the alcohol may be methanol, ethanol, 2-propanol, ethylene glycol, propylene glycol, or diethylene glycol monoethyl ether, and the ketone may be acetone, cyclohexanone, isophorone, or methyl ethyl ketone, and the aromatic The hydrocarbon may be benzene, toluene, xylene, ethylbenzene or methylnaphthalene, the aliphatic hydrocarbon may be hexane, cyclohexane, kerosene or light oil, the ester may be ethyl acetate or butyl acetate, and the nitrile may be It may be acetonitrile or isobutylnitrile, the ether may be diisopropyl ether or dioxane, the amide may be N,N-dimethylformamide or N.N-dimethylacetamide, and the halogenated hydrocarbon may be dichloromethane. , trichloroethane or carbon tetrachloride, and the vegetable oil may be soybean oil or cottonseed oil.

In addition, the solid carrier is a material for producing fine powder or granules, and may be one or two or more selected from minerals, natural organic materials, synthetic organic materials, salts, and synthetic inorganic materials, but is not limited thereto.

Specifically, the mineral may be kaolin clay, attapulgite clay, bentonite, montmorillonite, acid white clay, phyllophyllite, talc, diatomaceous earth, or talcite, and the natural organic material may be corn leaf powder or wall nut shell. It may be a powder, the synthetic organic material may be urea, the salt may be calcium carbonate or ammonium sulfate, and the synthetic inorganic material may be synthetic hydrated silicon oxide.

The surfactant may be one or two or more selected from anionic surfactants, nonionic surfactants, and cationic surfactants, but is not limited thereto.

Specifically, the anionic surfactant is an alkyl sulfate ester salt, alkylaryl sulfonate salt, dialkyl sulfosuccinate salt, polyoxyethylene alkylaryl ether phosphate ester salt, lignosulfonate salt, or naphthalene sulfonate formaldehyde condensation. It may be a compound, the nonionic surfactant may be polyoxyethylene alkyl aryl ether, polyoxyethylene alkylpolyoxypropylene block copolymer, or sorbitan fatty acid ester, and the cationic surfactant may be an alkyltrimethylammonium salt.

The auxiliary agent may include one or two or more selected from water-soluble polymers, polysaccharides, inorganic substances, preservatives, colorants, and stabilizers, but is not limited thereto.

Specifically, the water-soluble polymer may be polyvinyl alcohol or polyvinylpyrrolidone, the polysaccharide may be gum arabic, algilic acid and its salts, CMC (carboxymethyl-cellulose), or xanthan gum, and the inorganic material may be aluminum magnesium silicate or It may be an alumina sol. Additionally, in more detail, the stabilizer may be PAP (isopropyl acid phosphate) or BHT (butylhydroxytoluene).

In one embodiment, the composition for controlling plant diseases further includes one or two or more selected from the above-described carriers, surfactants, and auxiliaries to form an emulsion, emulsion, fluidizing agent, wettable powder, granulated wettable powder, powder, It can be formulated into granules, etc.

In addition, the present invention provides a method for producing the composition for controlling plant diseases, and the method for producing the composition for controlling plant diseases according to the present invention is culturing the Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain. It includes forming a culture medium.

In one embodiment, the culture medium may be prepared by inoculating Aspergillus Montenegroi SFC20200425-M27 strain into a medium and culturing with shaking, but is not limited thereto.

Shaking culture refers to a method of inoculating bacteria into a liquid medium and culturing it while continuously shaking it on a shaker. As a result, contact with medium components and oxygen supply are improved, so that proliferation can be uniform and efficiency can be improved.

The medium may be potato dextrose broth (PDB); 4 g of potato starch, 20 g of glucose, 1 L of distilled water), but is not limited thereto.

The shaking culture may be performed at 15 to 35°C at 90 to 210 rpm, and specifically may be performed at 20 to 30°C at 120 to 180 rpm, but is not limited thereto.

The shaking culture period may be 5 to 20 days, specifically 7 to 14 days, but is not limited thereto.

The method for producing a composition for controlling plant diseases according to an embodiment may further include removing the strain from the culture medium to prepare a culture supernatant.

The culture supernatant may be a filtrate obtained by centrifuging the culture medium in which the strain was cultured or filtering it with gauze or filter paper, that is, physically filtering it.

The method for producing a composition for controlling plant diseases according to an embodiment may further include the step of preparing an extract of the culture supernatant by treating the culture supernatant with an extraction solvent, and the extraction solvent may be ethyl acetate or butanol. It is not limited.

The ethyl acetate extract and butanol extract of the culture supernatant both show excellent plant disease control effects.

The manufacturing method can be performed under mild conditions and in a simple manner, making it easy to apply in industry.

Additionally, the present invention provides a method for controlling plant diseases using the composition for controlling plant diseases.

The plant disease control method according to one embodiment may include treating the plant, its seeds, or its habitat with the plant disease control composition.

The plant disease control method may include a plant disease control method that is a direct or indirect spray treatment method, a volatilization treatment method, or a seed treatment method. The composition for controlling plant diseases according to one embodiment can exhibit an easier and higher control effect using the above plant disease control method.

Specifically, the spraying method refers to a direct spraying method in which the composition for controlling plant diseases according to an embodiment is directly sprayed on plants, or an indirect spraying method in which the composition is sprayed on the soil in which the plants are growing or sprayed on a medium for cultivating plants. These control methods include treatment of areas where plants grow such as soil, treatment of plant stems and leaves, treatment of seeds such as seed sterilization or seed coating, and treatment of roots.

More specifically, the treatment of stems and leaves may include application to the plant surface, such as spraying the stems and leaves.

Also, more specifically, the treatment of the soil may include irrigation with a liquid treatment system, injection into the soil, or dripping of a liquid treatment agent, which is a method of spraying on the soil, mixing with the soil, or spreading the liquid treatment system into the soil. Examples of locations to be treated include planting holes, furrows, around planting holes, around planting furrows, the entire surface of the growing area, the area between the soil and the plant, the area between the roots, the area below the stem of the plant, and the main area. It may be a furrow, growing soil, bed, seedling box, seedling tray or seedbed.

In more detail, the treatment of the soil may be performed before spraying, during spraying, immediately after spraying, during a simulated cultivation period, before cultivation establishment, during cultivation establishment, or during the growth period after cultivation establishment. In the treatment of the soil, the active ingredient may be applied to the plant simultaneously, and a solid fertilizer such as a paste fertilizer containing the active ingredient may be applied to the soil. The active ingredients may be mixed in the irrigation liquid, injected into irrigation equipment such as irrigation tubes, irrigation pipes or sprinklers, mixed in the liquid that floods between furrows, or mixed in a water culture medium. Alternatively, the irrigation liquid and active ingredients can be premixed and used for treatment by any suitable irrigation method, including the irrigation methods mentioned above and other methods such as spraying and flooding.

Specifically, the volatilization method involves spraying a plant disease control composition according to an embodiment on media such as soil for cultivating plants, hydroponic media for culturing plants, and seedbeds, thereby protecting plants from pests and diseases through volatilization of the sprayed composition. This is a method of protecting the plant, and in addition, the plant can be exposed to the volatilized gaseous composition by placing the composition around the plant.

In addition, specifically, the seed treatment method includes a spray treatment method of spraying a suspension of a composition for controlling plant diseases according to an embodiment of the present invention on the seed surface, and a wettable powder, emulsion or fluidizing agent of the composition for controlling plant diseases according to an embodiment of the present invention. It may be a spray treatment applied on the seed surface by itself or by adding a small amount of water. In addition, the seed treatment method may be an impregnation treatment method, a film coating treatment method, or a pellet coating treatment method in which seeds are impregnated in a solution of a plant disease control composition according to an embodiment for a specific period of time.

When plants or soil for plant growth are treated with a composition for controlling plant diseases according to one embodiment, the treatment amount depends on the type of plant to be treated, the type and frequency of occurrence of pests to be controlled, formulation type, treatment period, or climatic conditions. It can change.

Emulsions, wettable powders, fluidizing agents, etc. are usually diluted with water and then sprayed for treatment. In this case, the concentration of the active ingredient is usually in the range of 0.0001 to 3% by weight, preferably 0.0005 to 1% by weight. Powders, granules, etc. are usually used for processing without dilution.

The plant disease control method according to one embodiment can be used in cultivated land such as rice fields or non-cultivated land.

Hereinafter, the composition for controlling plant diseases according to the present invention, its production method, and its control method will be described in more detail through specific examples. However, the following examples are only a reference for explaining the present invention in detail, and the present invention is not limited thereto, and may be implemented in various forms. Additionally, the terms used in the description in the present invention are only intended to effectively describe specific embodiments and are not intended to limit the present invention.

[Example 1] Isolation of strain SFC20150402-M24

10 g of soil collected from the Suncheon Bay mudflat (Haksin-ri, Byeolryang-myeon, Suncheon-si, Jeollanam-do) was suspended in 10 mL of artificial seawater (0.48 M NaCl, 0.01 M KCl, 0.027 M MgCl ₂ , 0.03 M MgSO ₄ , 0.01 M CaCl ₂ , NaHCO ₃ ). 100 μL of the suspension was spread on potato dextrose agar (4 g of potato starch, 20 g of glucose, 15 g of agar, 1 L of distilled water) and cultured at 25°C for 1 week.

The formed flora were transferred to a new potato agar medium and cultured to obtain strain SFC20200425-M27. Afterwards, the SFC20200425-M27 strain was immersed in a sterilized 20% glycerol aqueous solution and stored at -80°C.

Figure 1 shows the flora of Aspergillus montenegroi SFC20200425-M27 strain cultured on potato agar medium.

[Example 2] Molecular phylogenetic analysis of strain SFC20200425-M27

The SFC20200425-M27 strain isolated in Example 1 was inoculated into potato dextrose broth (4 g of potato starch, 20 g of glucose, 1 L of distilled water) and cultured with shaking at 150 r/min for 1 week at 25°C. Genomic DNA was extracted from the cultured bacteria using the AccuPrep DNA extraction kit (Bioneer, Daejeon, Korea), and PCR (polymerase chain reaction) was performed using the extracted DNA as a template.

PCR was performed once at 94°C for 5 minutes using forward primer CF1D (5′-CAGGTCTCCGAGTACAAG-3′), reverse primer CF4 (5′-CAGGTCTCCGAGTACAAG TTTYTGCATCATRAGYTGGAC-3′), and Ex Taq polymerase (Takara, Otsu, Japan). It was carried out under the following conditions: 30 seconds at 94°C, 30 seconds at 58°C, 30 seconds at 72°C, repeated 34 times, and once at 72°C for 7 minutes. The obtained PCR product was electrophoresed and purified using the Expin PCR purification kit (GeneAll, Seoul, Korea), and then a sequencing analysis service (Macrogen, Daejeon, Korea) was requested to analyze the base sequence of the calmodulin gene (CaM). .

Based on the sequenced CaM base sequence result (SEQ ID NO: 1), it was compared and analyzed with the CaM base sequences of other strains registered in the NCBI BLASTn database. After aligning the base sequences with the MAFFT program, a phylogenetic tree (neighbor-joining tree) was created using the MEGA program using the Kimura 2-parameter model and 1,000 bootstrap methods.

As a result of creating a phylogenetic tree using the sequence number, the SFC20200425-M27 strain had high homology (more than 99%) with the CBM-FA-669 and DTO 269C3 strains belonging to Aspergillus montenegroi . Figure 2 shows the phylogenetic tree.

Based on these analysis results, the SFC20200425-M27 strain was named Aspergillus montenegroi SFC20200425-M27 strain, and was deposited with the Agricultural Genetic Resources Center (KACC) of the Rural Development Administration under the accession number KACC 83033BP on July 31, 2020. did.

[Example 3] Preparation of culture supernatant of strain SFC20200425-M27

Aspergillus montenegroi ( Aspergillus montenegroi ) SFC20200425-M27 strain according to Example 1 was cultured on potato dextrose agar (4 g of potato starch, 20 g of glucose, 15 g of agar, 1 L of distilled water), and then the formed flora were corked. A mycelial disk (8 mm in diameter) was prepared by drilling. The mycelial disc was inoculated into potato dextrose broth (4g of potato starch, 20g of glucose, 1L of distilled water) and cultured at 25°C with shaking at 150r/min for 1 week to prepare a culture medium. The culture was centrifuged (10,000 × g, 20 minutes) and filtered through filter paper to prepare a culture supernatant (900 mL) (see S1 in Figure 3).

[Experimental Example 1] Plant disease control effect of culture supernatant of strain SFC20200425-M27

0.02 g of Tween 20 was added as a spreading agent to 80 mL of the culture supernatant according to Example 3, sprayed on the leaves of rice, tomato, wheat, and pepper seedlings, air-dried for 24 hours, and then inoculated with four types of pathogens. did. The four pathogens are rice blast ( Magnaporthe oryzae ), tomato blight ( Phytophthora infestans ), wheat red rust ( Puccinia triticina ), and pepper anthracnose ( Colletotrichum coccodes ). At this time, 4 pots were used for each plant bottle, and each plant was filled with about 70% of water supply or horticultural topsoil in a plastic pot with a diameter of 4.5 cm, then seeds were sown and grown in a greenhouse at 20 to 30°C for 1 week. Those grown for up to 3 weeks were used.

The disease spot area ratio (%) was examined 3 days after inoculation for pepper anthracnose, 4 days after inoculation for tomato blight, 5 days after inoculation for rice blast, and 7 days after inoculation for wheat red rust. The control value (%) was obtained by substituting the measured disease spot area ratio (%) of each plant disease into the following equation. The untreated group was treated with 0.02 g of Tween 20 in 80 mL of distilled water without culture supernatant.

Control value (%) = [1 - (disease area ratio of treated group / disease spot area rate of untreated group)] ×100

sample Control price (%) RCB ^a TLB WLR PAN Culture supernatant of strain SFC20200425-M27 33 98 100 35

^a RCB, rice blast; TLB, tomato late blight; WLR, wheat red rust; PAN, pepper anthracnose As shown in Table 1, the culture supernatant of the SFC20200425-M27 strain showed excellent control effects of 98% and 100% against tomato late blight and wheat red rust, respectively, and against rice blast and pepper anthracnose. showed a control effect of 33% and 35%, respectively. [Example 4] Preparation of extract of culture supernatant of SFC20150402-M24 strain

800 mL of culture supernatant of the Aspergillus montenegroi SFC20200425-M27 strain according to Example 3 and 800 mL of ethyl acetate were added to a separatory funnel, shaken, and allowed to stand to separate the upper layer. 800 mL of butanol was mixed again with the remaining lower layer, shaken, left to stand, and the upper layer was separated. These were each concentrated under reduced pressure and dried to finally obtain ethyl acetate extract (169 mg), butanol extract (169 mg), and water layer extract (1.5 g). (See S2 and S3 in Figure 3).

[Experimental Example 2] Plant disease control effect of extract of culture supernatant of SFC20150402-M24 strain

The ethyl acetate extract (169 mg), butanol extract (169 mg), and water layer extract (1.5 g) according to Example 4 were prepared at a concentration of 1,000 μg/mL, respectively, and the methanol content was added to 5% to dissolve the extract. Except for one thing, rice blast (pathogen: Magnaporthe oryzae ), tomato gray mold disease (pathogen: Botrytis cinerea ), tomato late blight (pathogen: Phytophthora infestans ), and wheat red rust (pathogen: Puccinia triticina ) were treated in the same manner as in Experimental Example 1. , barley powdery mildew (pathogen: Blumeria graminis f. sp. hordei ), and pepper anthracnose (pathogen: Colletotrichum coccodes ) were evaluated.

sample density
(μg/mL) Control price (%) RCB ^a TGM TLB WLR BPM PAN Ethyl acetate extract 1,000 79 7 84 100 58 85 butanol extract 1,000 85 0 82 100 8 85 water layer 1,000 0 0 0 0 0 6

^a RCB, rice blast; TGM, tomato gray mold disease; TLB, tomato late blight; WLR, wheat red rust; BPM, barley powdery mildew; PAN, pepper anthracnose As shown in Table 2, when plants were treated with ethyl acetate extract at a concentration of 1,000 μg/mL, it was 79% effective against rice blast, tomato blight, wheat red rust, barley powdery mildew, and pepper anthracnose, respectively. , showed excellent control effects of 84%, 100%, 58% and 85%. When butanol extract was applied to plants at a concentration of 1,000 μg/mL, it showed a control effect of 85%, 82%, 100%, and 85% against rice blast, tomato blight, wheat red rust, and pepper anthracnose, respectively. No significant effect was observed in the water layer, indicating that the active substance was a non-polar substance extracted with ethyl acetate and butanol. And there was no damage to rice, tomato, wheat, barley, and pepper crops treated with the extract.

National Academy of Agricultural Sciences Agricultural Genetic Resources Center (KACC) KACC83033BP 20200731

<110> KOREA RESEARCH INSTITUTE OF CHEMICAL TECHNOLOGY <120> Composition for controlling plant diseases using Aspergillus montenegroi SFC20200425-M27 strain and method for manufacturing the same <130> 20210614-10KR <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 505 <212> DNA <213> Artificial Sequence <220> <223> calmodulin gene (CaM) <400> 1 ctgtttgtaa ggccattggt tactgttata tcaaattcga atttatattg agagtatact 60 aatgtattcc gcacttaaca ggacaaggat ggcgatggtt agtgcatttg tccccccaga 120 cctgatcgca ttcgcccagc gtgtctgctg tagctctata taaaccgatt ctgataaacg 180 gcgacaggcc agattaccac taaggagctt ggcactgtca tgcgctcgct cggtcagaat 240 ccttcagagt ctgagcttca ggacatgatc aacgaagtgg acgccgacaa caatggcacc 300 attgactttc caggtacgcg aactttcagt ctacttcgca ccaagctaga aaccgtacta 360 atgctaaaca gagttcctta ccatgatggc cagaaagatg aaggacaccg attccgagga 420 ggaaattcgg gaggcgttca aggtcttcga ccgtgacaac aatggtttca tctccgctgc 480 tgagctgcgt cacgtcatga cctct 505

Claims (10)

Aspergillus Montenegroi SFC20200425-M27 strain deposited with accession number KACC 83033BP. A composition for controlling plant diseases comprising Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain, a culture fluid of the strain, a culture supernatant of the strain, an extract of the culture supernatant, or a mixture thereof as an active ingredient,
A composition for controlling plant diseases, wherein the plant disease is selected from rice blast, tomato blight, wheat red rust, barley powdery mildew, and pepper anthracnose. delete According to clause 2,
The composition for controlling plant diseases further comprises one or two or more selected from carriers, surfactants, and auxiliaries. A method for producing a composition for controlling plant diseases, comprising culturing the Aspergillus Montenegroi SFC20200425-M27 (Accession number: KACC 83033BP) strain to form a culture solution,
A method for producing a composition for controlling plant diseases, wherein the plant disease is selected from rice blast, tomato blight, wheat red rust, barley powdery mildew, and pepper anthracnose. According to clause 5,
A method for producing a composition for controlling plant diseases, further comprising the step of removing the strain from the culture medium to prepare a culture supernatant. According to clause 6,
A method for producing a composition for controlling plant diseases, further comprising the step of preparing an extract of the culture supernatant by treating the culture supernatant with an extraction solvent. According to clause 7,
A method of producing a composition for controlling plant diseases wherein the extraction solvent is ethyl acetate or butanol. A plant disease control method using the plant disease control composition of claim 2 or 4. According to clause 9,
The plant disease control method is a direct or indirect spraying method, a volatilization method, or a seed treatment method.