KR102375336B1 - Composition for controlling plant diseases using Brevibacillus brevis HK544 - Google Patents

Abstract

The present invention includes any one or more selected from the group consisting of Brevibacillus brevis HK544 strain, the culture medium of the strain, the culture filtrate of the strain, a fraction of the culture filtrate, or edeine B1 as an active ingredient To provide a composition for controlling plant diseases containing Since it exhibits an excellent control effect on diseases, it can be usefully used for controlling plant diseases.

Description

Composition for controlling plant diseases using Brevibacillus brevis HK544 strain

The present invention includes any one or more selected from the group consisting of Brevibacillus brevis HK544 strain, the culture medium of the strain, the culture filtrate of the strain, a fraction of the culture filtrate, or edeine B1 as an active ingredient To provide a composition for controlling plant diseases containing

Despite advances in science and technology and industry, the biggest challenge in modern agriculture is still solving the food problem. The world's population is rapidly increasing, and climate change is a red flag for a stable future food supply. Plant pathogens cause various diseases in crops and reduce the yield of crops. If plant diseases are not properly dealt with, serious social problems such as the Irish Great Famine of 1846 may occur. Chemical pesticides have been used as the main means for controlling plant diseases, but excessive use of chemical pesticides has brought about ecosystem disturbance, environmental pollution, human toxicity, and drug resistance problems. Therefore, the development of new eco-friendly plant disease control means to reduce or replace the use of chemical pesticides is required, and the need to develop eco-friendly non-toxic plant disease control technology using natural materials such as plant extracts, microorganisms, or microbial secondary metabolites this is getting bigger

Brevibacillus ( Brevibacillus spp.) is a rod-shaped Gram-positive bacterium, and through phylogenetic analysis based on the 16S rRNA gene sequence, it was reclassified from the Bacillus brevis cluster in 1996 to the genus Brevibacillus. . Brevibacillus agri ( Brevibaciilus agri ), Brevibacillus borstelensis , Brevibacillus borstelensis , Brevibacillus brevis ), Brevibacillus centrosporus ) choshinensis ), Brevibacillus formosus ( Brevibacillus formosus ), Brevibacillus laterosporus ( Brevibacillus laterosporus ), Brevibacillus parabrevis ( Brevibacillus parabrevis ), Brevibacillus reuszeri ( Brevibacillus reuszeri ), Currently, about 20 species of Brevibacillus exist, such as Brevibacillus thermoruber . Various antimicrobial peptides (eg, tostadin, edeine, spergualin, tauramamide, brevistin, Bac-GM100, laterosporulin, laterosporulin 10) was found in Brevibacillus spp.

Brevibacillus ( Brevibacillus spp.) is being used as a main material in research to search for new antibiotics. When tomato plants were treated with Fusarium oxysporum f. sp. lycopersici and Brevibacillus brevis strains at the same time, the progress of tomato wilt was greatly reduced, and the study results showed that the plant growth promoting effect was shown. (J Phytopathol 2010, 158: 470-478). In addition, the Brevibacillus brevis strain and the antibiotic gramicidin S produced by this strain showed excellent activity in inhibiting the spore germination and mycelial growth of the gray mold fungus ( Botrytis cinerea ), but plants The disease control effect is not suggested (Journal of Applied Microbiology 2001, 91, 652-659). In addition, Brevibacillus brevis ( Brevibacillus brevis ) FJAT-0809-GLX strain is a strain that produces an antibacterial active substance ethylparaben ( ethylparaben ), and the culture filtrate containing ethylparaben was treated with a plant as a plant pathogen ( Lasiodiplodia theobromae ). It has been reported that the occurrence of plant diseases in apple trees was greatly reduced. The above-mentioned antibacterial activity effects are not shown in all Brevibacillus ( Brevibacillus spp. ), and exhibit different antibacterial activity effects depending on the characteristics of the Brevibacillus spp. strain.

As such, Brevibacillus can be actively used to develop new crop protection agents because it produces various types of peptide antibiotics with antibacterial activity. The present inventors discovered and identified a Brevibacillus brevis HK544 strain showing a control effect on major plant diseases, and completed the present invention by identifying substances exhibiting antibacterial activity against plant pathogens from the culture solution.

It is an object of the present invention to provide a Brevibacillus brevis HK544 strain exhibiting plant disease control activity deposited with accession number KACC 81093BP.

Another object of the present invention is to provide a composition for controlling plant diseases containing as an active ingredient any one or more selected from the group consisting of a culture solution of the strain and a culture filtrate of the strain.

Another object of the present invention is to provide a plant disease control composition containing idein B1 as an active ingredient.

Another object of the present invention is Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain, a culture solution of the strain, a composition or ide for controlling plant diseases containing any one or more selected from the group consisting of the culture filtrate of the strain as an active ingredient It is to provide a method for controlling plant diseases comprising the step of treating plants or soil in which plants are grown with a plant disease controlling composition containing phosphorus B1 as an active ingredient.

In order to achieve the above object, an object of the present invention is to provide a Brevibacillus brevis HK544 strain exhibiting plant disease control activity deposited with accession number KACC 81093BP.

In order to achieve the above another object, the present invention provides a composition for controlling plant diseases containing, as an active ingredient, any one or more selected from the group consisting of a culture solution of the strain and a culture filtrate of the strain.

In order to achieve the above another object, the present invention provides a plant disease control composition containing idein B1 as an active ingredient.

In order to achieve the above another object, the present invention provides any one or more selected from the group consisting of Brevibacillus brevis HK544 strain, the culture medium of the strain, and the culture filtrate of the strain as an active ingredient. Plant disease control containing It provides a plant disease control method comprising the step of treating a plant or soil cultivating a plant with a plant disease control composition containing a composition for use or idein B1 as an active ingredient.

The composition for controlling plant diseases using the Brevibacillus brevis HK544 strain according to the present invention shows an excellent control effect on various plant diseases such as tomato gray mold, tomato late blight, wheat red rust, red pepper anthrax, etc. It can be usefully used for disease control purposes.

1 is a view showing the flora of the Brevibacillus brevis HK544 strain cultured in a soybean (tryptic soy agar) medium.
2 is a diagram showing a phylogenetic tree prepared by comparing the Brevibacillus brevis HK544 strain with related species based on the 16S rRNA gene nucleotide sequence.
Figure 3 is a photograph confirming the control effect of pepper anthrax of the Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain culture and the culture filtrate.
(A) From the left, no treatment group, 10% treatment group of HK544 culture solution, 20% treatment group of HK544 culture solution, 30% treatment group of HK544 culture solution, and group treated with synthetic pesticide dithianon (dithianon; 50 μg/ml)
(B) From the left, no treatment group, 10% treatment group of HK544 culture filtrate, 20% treatment group of HK544 culture filtrate, 30% treatment group of HK544 culture filtrate, treatment group of synthetic pesticide dithianon (50μg/ml)
4 is a view showing the process of separating the antibacterial active material edeine B1 (edeine B1) from the Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain.
5 is a diagram showing the results of ESI-MS cation analysis of edeine B1.
6 is a diagram showing the chemical structure of edeine B1 (edeine B1).

In the present specification, the term "control" includes the meaning of preventing and avoiding diseases or pests, as well as removing and killing them.

Hereinafter, the present invention will be described in detail.

The present invention provides a Brevibacillus brevis HK544 strain exhibiting plant disease control activity deposited with accession number KACC 81093BP.

The strain may have the nucleotide sequence of SEQ ID NO: 1.

The strain may be isolated from the soil, specifically, it may be isolated from the soil of the Korea Research Institute of Chemical Technology experimental field.

The strain may have plant disease control activity.

The plant disease is rice blast (causative bacterium: Magnaporthe oryzae ), tomato late blight ( causative bacterium: Phytophthora infestans ), wheat red rust ( causative bacterium: Puccinia triticina ), barley powdery mildew ( causative bacteria: Blumeria graminis f. sp. : Colletotrichum coccodes ), tomato gray mold disease (causative bacterium : Botrytis cinerea ), cabbage black spot disease (causative bacterium : Alternaria brassicicola ), phalaenopsis bacterial brown spot disease ( causative bacteria: Acidovorax avenae subsp. cattleyae ), peach bacterial hole disease (causative bacterium: Xanthomonas arboricola ) pv. pruni ), kiwi ulcer disease ( causative bacterium: Pseudomonas syringae pv. actinidiae ), or fruit tree burn disease ( causative bacterium: Erwinia amylovora ), but is not limited thereto.

In addition, the present invention provides a composition for controlling plant diseases containing, as an active ingredient, any one or more selected from the group consisting of a culture solution of the strain and a culture filtrate of the strain.

The strain may have the characteristics as described above as the Brevibacillus brevis HK544 strain deposited with the accession number KACC 81093BP.

The culture solution may be obtained by culturing the strain, and the culture filtrate may be a physical filtration of the culture solution culturing the strain.

The plant disease is rice blast (causative bacterium: Magnaporthe oryzae ), tomato late blight ( causative bacterium: Phytophthora infestans ), wheat red rust ( causative bacterium: Puccinia triticina ), barley powdery mildew ( causative bacteria: Blumeria graminis f. sp. : Colletotrichum coccodes ), tomato gray mold disease (causative bacterium : Botrytis cinerea ), cabbage black spot disease (causative bacterium : Alternaria brassicicola ), phalaenopsis bacterial brown spot disease ( causative bacteria: Acidovorax avenae subsp. cattleyae ), peach bacterial hole disease (causative bacterium: Xanthomonas arboricola ) pv. pruni ), kiwi ulcer disease ( causative bacterium: Pseudomonas syringae pv. actinidiae ), or fruit tree burn disease ( causative bacterium: Erwinia amylovora ), but is not limited thereto.

The plant may be any one or more selected from the group consisting of rice, tomato, barley, wheat, red pepper, Phalaenopsis, peach, kiwi, Chinese cabbage, apple tree and pear tree.

The composition for controlling plant diseases according to the present invention may be a simple mixture of the active material isolated from the Brevibacillus brevis HK544 strain, its culture medium, or its culture filtrate. Alternatively, the composition for controlling plant diseases is prepared by mixing the active material isolated from the culture medium or the culture filtrate with an inert carrier, and the mixture is an emulsion, emulsion, glidant, wettable powder, granulated wettable powder, powder, It is prepared by adding a surfactant and other necessary adjuvants to the mixture so that it can be formulated into granules or the like. The above-mentioned composition for controlling plant diseases by itself or by adding other inactive ingredients, the present invention can also be used as a seed treatment agent.

Examples of liquid carriers that can be used in the formulation include water; alcohols such as methanol and ethanol; ketones such as acetone and methyl ethyl ketone; aromatic hydrocarbons such as benzene, toluene, xylene, ethylbenzene and methyltaphthalene; aliphatic hydrocarbons such as hexane, cyclohexane, kerosine and light oil; esters such as ethyl acetate and butyl acetate; nitriles such as acetonitrile and isobutyrnitrile; ethers such as diisopropylether and dioxane; acid amides such as N,N-dimethyl formamide and N,N-dimethylacetamide; halogenated hydrocarbons such as dichloromethane, trichloroethane and carbon tetrachloride; dimethyl sulfoxide; and vegetable oils such as soybean oil and cottonseed oil.

Examples of solid carriers that can be used in the formulation include fine powders or granules such as minerals such as kaolin clay, attapulgite clay, bentonite, montmorillonite, acid white clay, phyllophyllite, talc, diatomaceous earth and talcite; natural organic substances such as corn leaf stalk powder and walnut hull powder; synthetic organic materials such as urea; salts such as calcium carbonate and ammonium sulfate; synthetic inorganic materials such as synthetic hydrated silicon oxide; As liquid carriers, aromatic hydrocarbons such as xylene, alkylbenzene and methylnaphthalene; alcohols such as 2-propanol, ethylene glycol, propylene glycol and ethylene glycol monoethyl ether; ketones such as acetone, cyclohexanone and isophorone; vegetable oils such as soybean oil and cottonseed oil; petroleum aliphatic hydrocarbons, esters, dimethylsulfoxide, acetonitrile and water.

Examples of surfactants include anionic surfactants such as alkyl sulfate ester salts, alkylaryl sulfonate salts, dialkylsulfosuccinate salts, polyoxyethylene alkylaryl ether phosphate ester salts, lignosulfonate salts and naphthalene sulfonate form aldehyde polycondensates; and nonionic surfactants such as polyoxyethylene alkyl aryl ethers, polyoxyethylene alkylpolyoxypropylene block copolymers and sorbitan fatty acid esters and cationic surfactants such as alkyltrimethylammonium salts.

Examples of other formulation aids include water-soluble polymers such as polyvinyl alcohol and polyvinylpyrrolidone, polysaccharides such as gum arabic, alginic acid and salts thereof, CMC (carboxymethyl-cellulose), xanthan gum, inorganic substances such as aluminum magnesium silicate and alumina sol (alumina sol), preservatives, colorants and stabilizers such as PAP (acid phosphate isopropyl) and BHT (butylhydroxyrtoluene).

In addition, the present invention is a step of treating at least one selected from the group consisting of Brevibacillus brevis HK544 strain deposited with accession number KACC 81093BP, its culture solution and its culture filtrate to a plant, its seed or its habitat. It provides a method for controlling plant diseases, characterized in that it comprises.

The treatment may be indirect spraying of spraying the composition directly on the plant, spraying on the soil in which the plant is growing, or spraying on the medium for culturing the plant.

The control method of the present invention includes treatment of stems and leaves of plants, treatment of places where plants are grown (eg soil), treatment of seeds such as seed sterilization/seed coating and treatment of roots.

As treatment of stems and leaves with the control method of the present invention, application on the surface of plants, such as, for example, spraying on stems and leaves, in particular can be included. Among the control methods of the present invention, the treatment of soil includes, for example, spraying onto the soil, mixing with the soil, and spraying the liquid treatment system into the soil (irrigation of the liquid treatment system, injection into the soil, dripping of the liquid treatment agent). Examples of places to be treated include planting holes, furrows, around planting furrows, around planting furrows, the entire surface of the growth site, between soil and plants, between roots, under the stems of plants Includes part, main furrow, growing soil, nail bed, seedling box, seedling tray, and seedbed. The treatment may be performed before spraying, during spraying, immediately after spraying, during the simulated cultivation period, before cultivation settlement, at the time of cultivation settlement, and during the growth period after cultivation settlement. In the above-mentioned soil treatment, the active ingredient may be simultaneously applied to the plant, or a solid fertilizer such as a paste fertilizer containing the active ingredient may be applied to the soil. The active ingredient may be mixed in an irrigation liquid, for example injected into an irrigation plant (irrigation tube, irrigation pipe, sprinkler, etc.), mixed in a liquid overflowing between furrows, or mixed in a water culture medium. can Alternatively, the irrigation liquid and active ingredient may be mixed beforehand and used for treatment by suitable irrigation methods, including, for example, the irrigation methods mentioned above and other methods such as spraying and flooding.

The volatilization treatment method as the control method of the present invention is, for example, the soil in which the plant is cultured with the composition for controlling plant diseases of the present invention, a hydroponic medium for culturing plants, and the volatilization of the sprayed composition. It is a method of protecting the plant from pests and diseases through the use of the composition, and in addition, the composition can be placed around the plant to expose the plant to the composition in the volatilized gaseous state.

The seed treatment method in the control method of the present invention is, for example, a method of treating seeds to be protected from pests with the composition for controlling plant diseases of the present invention, and a specific example thereof is a suspension of the composition for controlling plant diseases of the present invention in advance. a spray treatment method of securing and spraying on the seed surface; a spraying treatment method of applying the wettable powder, emulsion, glidant, etc. of the composition for controlling plant diseases of the present invention by itself or by adding a small amount of water to the seed surface; Impregnation treatment method of impregnating the seeds in the solution of the composition for controlling plant diseases of the present invention for a specific period; including film coating treatment and pellet coating treatment.

When a plant or soil for plant growth is treated with the compound according to the present invention, the treatment amount may vary depending on the type of plant to be treated, the type and frequency of occurrence of the pest to be controlled, the formulation form, the treatment period, climatic conditions, and the like.

Emulsions, wettable powders, glidants, etc. are usually diluted with water and then applied 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 treatment without dilution.

The control method of the present invention can be used in cultivated land or non-arable land, such as paddy fields.

The plant disease is rice blast (causative bacterium: Magnaporthe oryzae ), tomato late blight ( causative bacterium: Phytophthora infestans ), wheat red rust ( causative bacterium: Puccinia triticina ), barley powdery mildew ( causative bacteria: Blumeria graminis f. sp. : Colletotrichum coccodes ), tomato gray mold disease (causative bacterium : Botrytis cinerea ), cabbage black spot disease (causative bacterium : Alternaria brassicicola ), phalaenopsis bacterial brown spot disease ( causative bacteria: Acidovorax avenae subsp. cattleyae ), peach bacterial hole disease (causative bacterium: Xanthomonas arboricola ) pv. pruni ), kiwi ulcer disease ( causative bacterium: Pseudomonas syringae pv. actinidiae ), or fruit tree burn disease ( causative bacterium: Erwinia amylovora ), but is not limited thereto.

The plant may be any one or more selected from the group consisting of rice, tomato, barley, wheat, red pepper, Phalaenopsis, peach, kiwi, Chinese cabbage, apple tree and pear tree.

In a specific embodiment of the present invention, the present inventors identified the HK544 strain isolated from the soil of the experimental field at the Korea Research Institute of Chemical Technology and deposited it as a patent strain (accession number: KACC81093BP) at the Agricultural Genetic Resource Center of the Rural Development Administration (see Fig. 1).

In addition, the present inventors investigated the growth inhibitory activity against phytopathogenic fungi and bacteria after treating the plant with the culture filtrate of the Brevibacillus brevis HK544 strain of the present invention. , Tomato gray mold fungus ( Botrytis cinerea ), Red pepper anthrax ( Colletotrichum coccodes ), Phalaenopsis bacterial brown spot blight ( Acidovorax avenae subsp. cattleyae ), Peach bacterial hole bacillus ( Xanthomonas arboraicola pv. pruni ), Kiwi ulcer disease ( Pseudomonas arboricola pv. pruni ) actinidiae ) was confirmed to have a growth inhibitory effect (see Table 1).

In addition, the present inventors after treating the culture solution or the culture filtrate of the strain on plants, rice blast disease (pathogen: Magnaporthe oryzae ), tomato gray mold disease (pathogen: Botrytis cinerea ), tomato late blight (pathogen: Phytophthora infestans ), wheat red rust (Pathogen: Puccinia triticina ), barley powdery mildew ( Pathogen: Blumeria graminis f. sp. hordei ), pepper anthrax ( Pathogen: Colletotrichum coccodes ) As a result of confirming the plant disease control effect on, the culture medium or culture filtrate of the strain of the present invention is tomato It was confirmed that it has a control effect on gray mold disease, tomato late blight, wheat rust and red pepper anthrax (see Table 2).

In addition, the present invention provides a composition for controlling plant diseases containing a compound represented by the following formula (1), a stereoisomer thereof, as an active ingredient.

[Formula 1]

The chemical is edeine B1.

The compound may be isolated from Brevibacillus brevis HK544 strain, but is not limited thereto.

The plant disease is rice blast (causative bacterium: Magnaporthe oryzae ), tomato late blight ( causative bacterium: Phytophthora infestans ), wheat red rust ( causative bacterium: Puccinia triticina ), barley powdery mildew ( causative bacteria: Blumeria graminis f. sp. : Colletotrichum coccodes ), tomato gray mold disease ( causative bacterium : Botrytis cinerea ), cabbage black spot disease ( causative bacterium : Alternaria brassicicola ), phalaenopsis bacterial brown spot disease ( causative bacteria: Acidovorax avenae subsp. cattleyae ), peach bacterial hole disease (causative bacterium: Xanthomonas arboricola ) pv. pruni ), kiwi ulcer disease ( causative bacterium: Pseudomonas syringae pv. actinidiae ), or fruit tree burn disease ( causative bacterium: Erwinia amylovora ), but is not limited thereto.

The plant may be any one or more selected from the group consisting of rice, tomato, barley, wheat, red pepper, Phalaenopsis, peach, kiwi, Chinese cabbage, apple tree and pear tree.

The composition may have plant disease control activity.

The compound may be used in the form of an agrochemically acceptable salt, and may include all salts, hydrates and solvates that can be prepared by conventional methods as well as agrochemically acceptable salts.

In addition, the present invention provides a method for controlling plant diseases using a composition containing the compound as an active ingredient.

The treatment may be indirect spraying of spraying the composition directly on the plant, spraying on the soil in which the plant is growing, or spraying on the medium for culturing the plant.

The control method of the present invention includes treatment of stems and leaves of plants, treatment of places where plants are grown (eg soil), treatment of seeds such as seed sterilization/seed coating and treatment of roots.

As treatment of stems and leaves with the control method of the present invention, application on the surface of plants, such as, for example, spraying on stems and leaves, in particular can be included. Among the control methods of the present invention, the treatment of soil includes, for example, spraying onto the soil, mixing with the soil, and spraying the liquid treatment system into the soil (irrigation of the liquid treatment system, injection into the soil, dripping of the liquid treatment agent). Examples of places to be treated include planting holes, furrows, around planting furrows, around planting furrows, the entire surface of the growth site, between soil and plants, between roots, under the stems of plants Includes part, main furrow, growing soil, nail bed, seedling box, seedling tray, and seedbed. The treatment may be performed before spraying, during spraying, immediately after spraying, during the simulated cultivation period, before cultivation settlement, at the time of cultivation settlement, and during the growth period after cultivation settlement. In the above-mentioned soil treatment, the active ingredient may be simultaneously applied to the plant, or a solid fertilizer such as a paste fertilizer containing the active ingredient may be applied to the soil. The active ingredient may be mixed in an irrigation liquid, for example injected into an irrigation plant (irrigation tube, irrigation pipe, sprinkler, etc.), mixed in a liquid overflowing between furrows, or mixed in a water culture medium. can Alternatively, the irrigation liquid and active ingredient may be mixed beforehand and used for treatment by suitable irrigation methods including, for example, the irrigation methods mentioned above and other methods such as spraying and flooding.

The volatilization treatment method as the control method of the present invention is, for example, the soil in which the plant is cultured with the composition for controlling plant diseases of the present invention, a hydroponic medium for culturing plants, and the volatilization of the sprayed composition. It is a method of protecting the plant from pests and diseases through the use of the composition, and in addition, the composition can be placed around the plant to expose the plant to the composition in the volatilized gaseous state.

The seed treatment method in the control method of the present invention is, for example, a method of treating seeds so as to be protected from pests with the composition for controlling plant diseases of the present invention, and a specific example thereof is a suspension of the composition for controlling plant diseases of the present invention. a spray treatment method of spraying on the surface; a spraying treatment method of applying the wettable powder, emulsion, glidant, etc. of the composition for controlling plant diseases of the present invention by itself or by adding a small amount of water to the seed surface; Impregnation treatment method of impregnating the seeds in the solution of the composition for controlling plant diseases of the present invention for a specific period; including film coating treatment and pellet coating treatment.

When a plant or soil for plant growth is treated with the compound according to the present invention, the treatment amount may vary depending on the type of plant to be treated, the type and frequency of occurrence of the pest to be controlled, the formulation form, the treatment period, climatic conditions, and the like.

Emulsions, wettable powders, glidants, etc. are usually diluted with water and then applied 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 treatment without dilution.

The control method of the present invention can be used in cultivated land or non-arable land, such as paddy fields.

In a specific embodiment of the present invention, the present inventors separate the edeine B1 represented by Formula 1 from the fraction obtained by sequentially fractionating the culture filtrate of the Brevibacillus brevis HK544 strain with butanol and acetone. (see FIGS. 4 and 5).

In addition, the present inventors found that the isolated edeine B1 is a phytopathogenic mold cabbage black blight ( Alternaria brassicicola ), tomato gray mold blight ( Botrytis cinerea ), red pepper anthrax ( Colletotrichum coccodes ), rice blast bacillus ( Magnaporthe ) oryzae ), tomato late blight ( Phytophthora infestans ), and phytopathogenic bacteria Fruit burn ( Erwinia amylovora ) were confirmed to have an excellent bactericidal activity (see Table 4).

Hereinafter, the present invention will be described in detail by way of Examples.

However, the following examples only illustrate the present invention, and the scope of the present invention is not limited by the following examples.

<Example 1> Isolation of strain HK544

To isolate the HK544 strain, a soil sample was collected from the soil of the experimental field at the Korea Research Institute of Chemical Technology and mixed with water. After filtering the residues using a 25 µm sieve, 0.1 ml of the suspension is taken and 1/10 strength tryptic soy agar; 0.17% tryptone, 0.03% soytone, 0.025% dextrose, 0.05% NaCl, 0.25% After smearing on K ₂ HPO ₄ , 1.5% agar), it was cultured in an incubator at 25° C. for 3 days. The isolated bacteria obtained a single colony through three passages, and was named HK544 strain. Until used in subsequent experiments, the HK544 strain was immersed in a sterile 20% glycerol aqueous solution and stored at -80°.

<Example 2> Molecular biological identification of strain HK544

HK544 strain used in the present invention TSA (tryptic soy agar; 1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ , 1.5% agar) When cultured in medium, ivory-colored slime form of the colony (Fig. 1). Molecular phylogenetic analysis was performed based on the 16S rRNA gene nucleotide sequence of the HK544 strain. The HK544 strain was inoculated in TSA medium and cultured with shaking at 150 rpm for 2 days at 25°, using CTAB buffer, and then DNA of the KH544 strain (100 ng/μl) was obtained by using a DNA extraction method. Polymerase chain reaction using the DNA as a template and a pair of universal primer sets (27F, 5'-AGAGTTTGATCMTGGCTCAG-3'; 1492R, 5'-TACGGYTACCTTGTTACGACTT-3') and Ex Taq polymerase (Takara, Otsu, Japan) chain reaction) to obtain a 16S rRNA gene fragment. The band identified in 0.8% agarose gel was purified using the GeneAll Expin PCR purification kit (GeneAll, Seoul, Korea), and then sequencing analysis service (Macrogen, Daejeon, Korea) was used for sequencing analysis. did

As a result, the 16S rRNA gene sequence of SEQ ID NO: 1 was obtained (SEQ ID NO: 1), and the 16S rRNA gene sequence of other strains registered in the NCBI BLASTn (http://www.ncbi. nlm.nih.gov) database and comparative analysis. After aligning the nucleotide sequences with Clustal W using the MEGA 4.0 program, the similarity between genes was created as a neighbor-joining tree (Saitou and Nei, 1987). The molecular tree was constructed using the neighbor-joining method, one of the distance matrix methods, and Bootstrap analysis (1,000 times) was performed using the MEGA program algorithm.

[SEQ ID NO: 1]

AGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGAGTGTCTTCGGACCCTAGCGGCGGACGGGTGAGTAACACGTAGGCAACCTGCCTCTCAGACTGGGATAACATAGGGAAACTTATGCTAATACCGGATAGGTTTTTGGATCGCATGATCCGAAAAGAAAAGGCGGCTTCGGCTGTCACTGGGAGATGGGCCTGCGGCGCATTAGCTAGTTGGTGGGGTAACGGCCTACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATTTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAACGATGAAGGTCTTCGGATTGTAAAGTTCTGTTGTTAGGGACGAATAAGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTGACGAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGCGCGCAGGCGGCTATGTAAGTCTGGTGTTAAAGCCCGGAGCTCAACTCCGGTTCGCATCGGAAACTGTGTAGCTTGAGTGCAGAAGAGGAAAGCGGTATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGTCTGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTGGGGGTTTCAATACCCTCAGTGCCGCAGCTAACGCAATAAGCACTCCGCCTGGGGAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCGCTGACCGCTCTGGAGACAGAGCTTCCCTT CGGGGCAGCGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCTTTAGTTGCCAGCATTCAGTTGGGCACTCTAGAGAGACTGCCGTCGACAAGACGGAGGAAGGCGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGTTGGTACAACGGGATGCTACCTCGCGAGAGGACGCCAATCTCTTAAAACCAATCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGGGAGTTTGCAACACCCGAAGTCGGTGAGGTAACCGCAAGGAGCCAGCCGCCGAAGGTGGGGTAGATGACTGGGGTGAAGTCGTA

As a result of 16S rRNA gene sequencing, the HK544 strain formed the same group as the Brevibacillus brevis DSM 5760 strain (FIG. 2), and showed high homology (99.9% or more). Based on these results, the HK544 strain was identified as Brevibacillus brevis and deposited as a patent strain (accession number: KACC81093BP) at the Agricultural Genetic Resource Center of the Rural Development Administration.

<Example 3> Preparation of culture solution of HK544 strain, culture filtrate and dilutions thereof

Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain was cultured in TSA medium (1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ , 1.5% agar), and then on TSA agar medium. 200 ml of sterilized TSB medium (tryptic soy broth; tryptic soy agar; 1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ ) by sealing the formed flora with the face 1 L -Inoculated into an Erlenmeyer flask, and incubated with shaking (150 rpm) at 25˚C for 2 days to prepare a pre-culture solution. 1% of the pre-culture solution was inoculated in a 2 L-erlenmeyer flask containing 400 ml of fresh TSB medium, and cultured with shaking at 25 °C (150 rpm) for 3 days after inoculation to prepare a HK544 culture solution.

In addition, Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain culture solution was centrifuged (10,000 × g, 20 min) and filtered through filter paper (Whatman No. 1) to prepare a culture filtrate. Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain culture solution or a sample diluted with distilled water to 10%, 20%, 30% concentration was used for plant disease control test.

<Experimental Example 1> In vitro bactericidal activity effect of culture filtrate of HK544 strain

The Brevibacillus brevis HK544 strain culture filtrate prepared in Example 3 was used as a phytopathogenic fungus, Chinese cabbage black mold ( Alternaria brassicicola ), tomato gray mold ( Botrytis cinerea ), red pepper anthrax ( Colletotrichum coccodes ) , Rice blast bacillus ( Magnaporthe oryzae ), tomato blight blight ( Phytophthora infestans ) and phytopathogenic bacteria Phalaenopsis bacterium brown spot blight ( Acidovorax avenae subsp. cattleyae ), fruit root nodules ( Agroholderia glumaebacterium ), Bacterial rice germ , Clavibacter michiganensis subsp. michiganensis ), Pectobacterium carotovorum subsp. carotovorum ), Pseudomonas syringae pv . The minimum inhibitory concentration (MIC) was determined by microdilution in liquid medium using a 96-well plate.

In 100 μl wells, spores were incubated at a concentration of 1 × 10 ⁴ spores/ml for mold or 1 × 10 ³ baceterial cells/ml for bacteria, respectively. Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain culture filtrate was treated in each well so that the concentration of 1.3%, 2.5%, 5%, 10%, 20%. At this time, sterile water was used as an untreated instrument. When observed with the naked eye after culturing at the optimum temperature for microbial growth for 2-3 days, the concentration at which fungal growth was completely inhibited was determined as the minimum inhibitory concentration (MIC).

plant pathogens Minimum inhibitory concentration (%) Alternaria brassicicola 1.3 Botrytis cinerea 1.3 Colletotrichum coccodes 1.3 Magnaporthe oryzae >20 Phytophthora infestans >20 Acidovorax avenae subsp. cattleyae 10 Agrobacterium tumefaciens >20 Burkholderia glumae >20 Clavibacter michiganensis subsp. michiganensis >20 Pectobacterium carotovorum subsp. carotovorum >20 Dickya chrysanthemi >20 Pseudomonas syringae pv. lachrymans >20 Xanthomonas arboricola pv. pruni 20 Pseudomonas syringae pv. actinidiae 20 Ralstonia solanacearum >20

As a result, as shown in Table 1, the culture filtrate of the HK544 strain was at a concentration of 1.3%, the cabbage black pattern fungus ( Alternaria brassicicola ), the tomato gray mold fungus ( Botrytis cinerea ), the red pepper anthrax ( Colletotrichum coccodes ) in completely inhibiting It showed in vitro bactericidal activity. In addition, the culture filtrate of the HK544 strain at a concentration of 10-20% exhibits an activity of completely inhibiting the growth of three types of phytopathogenic bacteria ( A. avenae subsp. cattleyae , X. arboricola pv. pruni , P. syringae pv. actinidiae ). it was

<Experimental Example 2> Confirmation of plant disease control effect of HK544 strain culture solution, culture filtrate and dilutions thereof

The six major plant diseases, rice blast disease (pathogen: Magnaporthe oryzae ), tomato gray mold disease (pathogen: Botrytis cinerea ), tomato blight (pathogen: Phytophthora infestans ), wheat red rust (pathogen: Puccinia triticina ), barley powdery mildew (pathogen: Blumeria graminis f. sp. hordei ), pepper anthrax (pathogen: Colletotrichum coccodes ) The effect was confirmed under greenhouse conditions.

To evaluate the plant disease control effect, 0.025% (w/v) Tween 20 was added to the sample as an electrodeposition agent, and in this case, 0.025% (w/v) Tween 20 aqueous solution was used in the untreated group. The sample was spray-treated on rice, tomato, wheat, barley and red pepper seedlings, and each pathogen was inoculated one day later to examine the plant disease control activity of the culture medium or the culture filtrate.

Specifically, 4 pots were used for each plant bottle, and after spraying the sample on the leaf surface, it was air-dried for 24 hours, and then each plant pathogen was inoculated. The rice, tomato, barley and wheat plants used in the experiment were filled with about 70% of water or horticultural tops in a plastic pot with a diameter of 4.5 cm, and then seeds were sown and grown in a greenhouse at 25° C. for 1 to 4 weeks.

Tomato gray mold and tomato late blight were investigated 3 days after inoculation, rice blast disease 5 days after inoculation, and wheat red rust and barley powdery mildew 7 days after inoculation. For the control activity experiment on red pepper anthracnose, it was developed in a humid room for 2 days and then left in a constant temperature room at 25 ° C for 1 day. %) was obtained.

Control value (%) = [1 - (Ratio of lesions in treated areas / Ratio of lesions in untreated areas)] ×100

density(%) disease prevention value (%) RCB ^a TGM TLB WLR BPM PAN culture medium 10 14 99 92 60 0 63 20 14 100 87 43 0 58 30 14 96 84 80 17 92 culture filtrate 10 0 97 95 0 0 47 20 0 98 95 10 0 79 30 0 98 95 53 0 82 ^a RCB, rice blast; TGM, tomato gray mold; TLB, tomato blight; WLR, wheat red rust; BPM,
barley powdery mildew; PAN, red pepper anthrax

As a result, as shown in Table 2, the culture solution or the culture filtrate of the HK544 strain showed excellent control activity in controlling more than 90% of tomato gray mold disease and tomato late blight even at a concentration of 10%, and against wheat red rust and red pepper anthrax. showed inhibitory activity. The culture solution or culture filtrate of the HK544 strain showed an effect of controlling more than 80% of red pepper anthrax at a concentration of 30%, which was equivalent to the effect of treating the synthetic pesticide dithianon at a concentration of 50 μg/ml (Fig. 3) ). In addition, the culture medium and the culture filtrate of the HK544 strain showed an effect of controlling 80% and 53% of wheat red rust, respectively, at a concentration of 30%.

<Example 4> Separation of bactericidal active substances from the culture filtrate of strain HK544

Brevibacillus brevis HK544 strain was cultured in TSA medium (1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ , 1.5% agar), and then formed in TSA medium 1 L triangle containing 200 ml of TSB medium (tryptic soy broth; tryptic soy agar; 1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ ) sterilized by sealing the flora A preculture was prepared by inoculating the flask and culturing with shaking (150 rpm) at 25 ˚C for 2 days. 1% of the pre-culture solution was inoculated into 10 2 L Erlenmeyer flasks containing 400 ml of new TSB medium, and cultured with shaking (150 rpm) at 25 ° C for 3 days after inoculation to prepare a HK544 culture solution (4 L). Thereafter, the Brevibacillus brevis ( Brevibacillus brevis ) HK544 strain culture solution was centrifuged (10,000 × g, 20 min) and filtered through filter paper (Whatman No. 1) to prepare a culture filtrate.

The HK544 culture filtrate (4 L) was loaded into a column ( Φ 8 cm × 40 cm) filled with polystyrene resin (Diaion HP-20, Mitsubishi Chemicals), and 20%, 40%, 60%, 80%, 100 6 fractions of F1-F6 were prepared by eluting 1 L sequentially with % aqueous methanol solution, and finally eluting with 100% acetone (1 l), and each fraction was added to the pepper anthrax at a concentration of 5% ( Colletotrichum coccodes ) When treated, it was confirmed that the F4 fraction showed strong bactericidal activity.

The F4 fraction (2 g) was loaded into a column (Φ 4 cm × 20 cm) filled with C18 resin (LiChroprep RP-18, Merck) and 20%, 40%, 50%, 60%, 70% using a vacuum pump. Seven fractions of F41-F47 were prepared by eluting with %, 90%, 100% aqueous methanol solution (including 0.1% formic acid), and when each fraction was treated with red pepper anthrax at a concentration of 200 μg/ml, the F41 fraction It was confirmed that strong bactericidal activity appears in

The F41 fraction (200 mg) was dissolved in a small amount of water and loaded onto a column (XK16/100, GE Healthcare) filled with a cross-linked dextran resin (Sephadex LH-20, Merck), and 0.1 ml per minute. The fractions that were passed through water at a flow rate were fractionated by 2 ml each, and when each fraction was treated at a concentration of 25 μg/ml, it was confirmed that the activity appeared in fractions 58-62.

¹ H-NMR spectrum of fractions 58-62 was confirmed to be the same, and 117 mg of compound 1 was purified by combining them. Brevibacillus brevis ( Brevibacillus brevis ) The separation and purification sequence of the bactericidal active material from the culture filtrate of the HK544 strain is shown in FIG. 4 .

<Example 5> Chemical structure identification of bactericidal active substances isolated from HK544 culture filtrate

In order to perform MS and NMR analysis to determine the chemical structure of Compound 1, Compound 1 was dissolved in D ₂ O (99.9 atom% D; Cambridge Isotope Laboratories) and ¹ H- and ¹³ C- NMR spectra and COSY, HSQC, and HMBC spectra were analyzed. And the molecular weight of the compound was measured using an ESI-MS mass spectrometer (Waters).

carbon chemical shift (ppm) δ _C ,type δ _H ( J inHz) One 51.5, CH 4.64 dt (7.5, 3.9) 2 56.4, CH ₂ 2.92 ddd (15.1, 8.2, 3.0); 3.0 m 3 172.5, CO 4 43.3, CH ₂ 3.27 dt (6.7, 2.0); 3.44 m 5 71.2, CH 4.09 m 6 175.9, CO 7 48.2, CH 4.71 dt (8.1, 4.0) 8 170.3, CO 9 52.8, CH 4.14 m 10 32.1, CH ₂ 1.56 m; 1.69 m 11 22.6, CH ₂ 1.26 m; 1.38 m 12 27.6, CH ₂ 1.52 m; 1.76 m 13 56.2, CH 3.27 dt (6.7, 2.0) 14 68.5, CH 4.29 ddt (10.2, 6.9, 3.6) 15 39.0, CH ₂ 2.42 m 16 174.4, CO 17 43.5, CH ₂ 3.87 d (2.5) 18 172.8, CO 19 37.1, CH ₂ 3.18 t (6.8) 20 26.6, CH ₂ 1.60 m; 1.76 m 21 40.1, CH ₂ 3.0 m 22 43.7, CH ₂ 3.0 m 23 26.1, CH ₂ 1.52 m; 1.60 m 24 23.9, CH ₂ 1.52 m; 1.60 m 25 46.0, CH ₂ 3.0 m 26 157.5, C 27 41.5, CH ₂ 3.34 dq (7.5, 3.7); 3.35 dq (14.5, 5.5) 28 179.4, CO One' 127.9, C 2',6' 129.8, (CH) ₂ 8.55 d (8.5) 3',5' 117.0, (CH) ₂ 6.9 d (8.5) 4' 157.8, C

As a result, compound 1 was detected as a cation at m/z 797 [M + H]+, m/z 819 [M + Na] ⁺ , m/z 836 [M + K] ⁺ by ESIMS, and the molecular weight was MW 796 was determined as (FIG. 5). As shown in Table 3, in the ¹³ C-NMR spectrum of Compound 1, six carbonyl carbons ( δ _C 179.4, 175.9, 174.4, 172.8, 172.5, 170.3, 157.8), three quaternary sp ² ( δ _C 157.8, 157.5, 127.9), 4 methine sp ² ( δ _C 129.8, 117.0; superimposed), 6 methine sp ³ ( δ _C 71.2, 68.5, 56.2, 52.8, 51.5, 48.2), 15 methylene sp ² ( δ _C 56.4) , 46.0, 43.7, 43.5, 43.3, 41.5, 40.1, 39.0, 37.1, 32.1, 27.6, 26.6, 26.1, 23.9, 22.6) signals (Table 3). The position of the hydrogen signal was determined through 1H-1H COSY, HSQC, and HMBC analysis, and by synthesizing the ¹ H- and ¹³ C-NMR spectra in Table 3 and literature data, the chemical structure of Compound 1 was determined as the linear peptide antibiotic idein. B1 (edeine B1) was determined (FIG. 6).

<Experimental Example 3> Edein B1 (edeine B1) for various plant pathogens in vitro bactericidal activity

Phytopathogenic fungi of edeine B1 ( Alternaria brassicicola ), tomato gray mold ( Botrytis cinerea ), red pepper anthrax ( Colletotrichum coccodes ), rice blast bacillus ( Magnaporthe oryzae ), tomato blight ( Phytophthora ) infestans ) and the phytopathogenic bacterium, Erwinia amylovora , the growth inhibitory activity against the 96-well plate was used to determine the minimum inhibitory concentration (MIC) by microdilution method.

Incubate in 100 μl well to contain spores at a concentration of 1 Х10 ⁴ spores/ml for mold or 1 Х10 ³ bacterial cells/ml for bacteria, and edeine B1 is 0.1, 0.2, 0.4, 0.8, respectively. , 1.6, 3.1, 6.3, 12.5, 25, 50, 100, and 200 μg/ml were prepared. At this time, the methanol content was less than 1%, and 1% methanol was used as an untreated group, and the antibiotics cycloheximide and oxytetracycline were used as positive controls for phytopathogenic fungi and bacteria, respectively. When observed with the naked eye after culturing at the optimum temperature for microbial growth for 2-3 days, the concentration at which the growth of mold and bacteria was completely inhibited was determined as the minimum inhibitory concentration (MIC).

phytopathogenic microorganisms Minimum inhibitory concentration (μg/ml) Edein B1 cyclo
Hexamide oxytetra
cyclin mold Alternaria brassicicola 0.4 12.5 na Botrytis cinerea 1.6 100 na Colletotrichum coccodes 12.5 50 na Magnaporthe oryzae 12.5 1.6 na Phytophthora infestans 0.1 0.2 na Germ Erwinia amylovora 1.6 na 1.6 na, not analyzed

As a result, as shown in Table 4, edeine B1 (edeine B1) exhibited a wide range of antibacterial activity against various phytopathogenic fungi. Specifically, edeine B1 (edeine B1) at a concentration of 0.4 μg/ml of a cabbage black pattern blight ( Alternaria brassicicola ), a tomato gray mold blight ( Botrytis cinerea ) at a concentration of 1.6 μg/ml, Colletotrichum coccodes ) at a concentration of 12.5 μg/ml, rice blast bacillus ( Magnaporthe oryzae ) at a concentration of 12.5 μg/ml, and tomato late blight ( Phytophthora infestans ) at a concentration of 0.1 μg/ml. This indicates that the effect is better than cycloheximide, a natural antibiotic derived from actinomycetes. In addition, edeine B1 showed bactericidal activity to completely inhibit Erwinia amylovora , a phytopathogenic bacterium, at a concentration of 1.6 μg/ml, which is similar to that of the antibiotic oxytetracycline. show the effect.

<Experimental Example 4> Comparison of bactericidal activity between strains HK544 and closely related strains

Brevibacillus brevis ATCC 8186, Brevibacillus brevis ATCC 8185 , Brevibacillus brevis ( Brevibacillus brevis ) DSM 30, Brevibacillus brevis ( Brevibacillus reuszeri ) DSM 9887, Brevibacillus sp. FMR09-05, Brevibacillus panachihumi DCY35, Brevibacillus borstelensis DSM 6347, Brevibacillus parabrevis ) NBRC 12334, Brevibacillus joshinensis ( Brevibacillus choshinensis ) DSM 8552, Brevibacillus agri ( Brevibacillus agri ) DSM 6348, Brevibacillus formosus ( Brevibacillus formosus ) DSM 9885 was sold.

The received strain was cultured in TSA (tryptic soy agar; 1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ , 1.5% agar) medium, and then a single colony formed on an agar medium 250 ml Erlenmeyer flask containing 50 ml of sterile TSB medium (tryptic soy broth; tryptic soy agar; 1.7% tryptone, 0.3% soytone, 0.25% dextrose, 0.5% NaCl, 2.5% K ₂ HPO ₄ ) sealed with face A culture solution was prepared by inoculating it in a cell, shaking culture at 25 ˚C for 3 days (150 rpm), centrifugation (10,000 × g, 20 min), and then filtering with filter paper (Whatman No. 1) to prepare a culture filtrate. Using YPG medium (1% yeast extract, 2% peptone, 2% glucose) added to the culture filtrate of Brevibacillus sp. at a concentration of 5% or 10% in a 96-well plate, yeast ( Saccharomyces cerevisiae ) The antibacterial activity of the Brevibacillus sp. strain was evaluated. Sterilized distilled water was used for the untreated section.

YPG medium (5 ml) was inoculated with yeast ( Saccharomyces cerevisiae ) colonies, and yeast bacteria cultured with shaking (150 rpm) for 1 day at 30 ° was used as an inoculum, and when measured with a UV/Vis spectrometer, 600 nm It was diluted with sterilized distilled water to have an absorbance of 0.3. The yeast suspension was inoculated in a 96-well plate at a concentration of 1%. After inoculation of yeast, cultured at 30° for 1 day, absorbance at 600 nm was measured to investigate the growth of yeast. By substituting the following formula, the growth inhibition rate (%) of yeast was investigated.

Growth inhibition rate (%) = (1 - OD ₆₀₀ value of treated group / OD ₆₀₀ value of untreated group) ×100

test microorganism Culture filtrate concentration (%) Growth inhibition for yeast (%) B. brevis HK544 10 93 ±3 5 6 ±1 B. brevis ATCC8186 10 1 ±1 5 2 ±1 B. brevis ATCC8185 10 1 ±0 5 1 ±1 B. brevis DSM30 10 0 5 1 ±1 B. reuszeri DSM9887 10 0 5 1 ±1 Brevibacillus sp .FMR09-05 10 3 ±0 5 4 ±1 B. panacihumi DCY35 10 1 ±1 5 4 ±1 B. borstelensis DSM6347 10 0 5 2 ±2 B. parabrevis NBRC12334 10 1 ±0 5 3 ±1 B. choshinensis DSM8552 10 1 ±0 5 2 ±1 B. agri DSM6348 10 0 5 2 ±2 B. formosus DSM9885 10 0 5 0

As a result, as shown in Table 5, the sterilization activity of the culture filtrate of the Brevibacillus brevis HK544 strain among the 12 types of tested Brevibacillus culture filtrate was the best. When the culture filtrate of the HK544 strain was treated at a concentration of 10%, the growth of yeast was inhibited by 93% compared to the untreated group, but other strains had no effect of inhibiting the growth of yeast.

Brevibacillus of other species as well as culture filtrates of the same species ATCC 8186, ATCC8185, DSM 30 did not show bactericidal activity. This suggests that the sterilization mechanism of the Brevibacillus brevis HK544 strain is a unique characteristic of the HK544 strain appearing at the individual (strain or isolate) level, rather than a characteristic commonly seen in the Brevibacillus brevis species.

<110> KRICT <120> Composition for controlling plant diseases using Brevibacillus brevis HK544 <130> 2020p-06-037 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1468 <212> DNA <213> Artificial Sequence <220> <223> Brevibacillus brevis HK544 DNA sequence <400> 1 aggacgaacg ctggcggcgt gcctaataca tgcaagtcga gcgagtgtct tcggacccta 60 gcggcggacg ggtgagtaac acgtaggcaa cctgcctctc agactgggat aacataggga 120 aacttatgct aataccggat aggtttttgg atcgcatgat ccgaaaagaa aaggcggctt 180 cggctgtcac tgggagatgg gcctgcggcg cattagctag ttggtggggt aacggcctac 240 caaggcgacg atgcgtagcc gacctgagag ggtgaccggc cacactggga ctgagacacg 300 gcccagactc ctacgggagg cagcagtagg gaattttcca caatggacga aagtctgatg 360 gagcaacgcc gcgtgaacga tgaaggtctt cggattgtaa agttctgttg ttagggacga 420 ataagtaccg ttcgaatagg gcggtacctt gacggtacct gacgagaaag ccacggctaa 480 ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggat ttattgggcg 540 taaagcgcgc gcaggcggct atgtaagtct ggtgttaaag cccggagctc aactccggtt 600 cgcatcggaa actgtgtagc ttgagtgcag aagaggaaag cggtattcca cgtgtagcgg 660 tgaaatgcgt agagatgtgg aggaacacca gtggcgaagg cggctttctg gtctgtaact 720 gacgctgagg cgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc 780 gtaaacgatg agtgctaggt gttgggggtt tcaataccct cagtgccgca gctaacgcaa 840 taagcactcc gcctggggag tacgctcgca agagtgaaac tcaaaggaat tgacgggggc 900 ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 960 ttgacatccc gctgaccgct ctggagacag agcttccctt cggggcagcg gtgacaggtg 1020 gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080 acccttatct ttagttgcca gcattcagtt gggcactcta gagagactgc cgtcgacaag 1140 acggaggaag gcggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1200 tgctacaatg gttggtacaa cgggatgcta cctcgcgaga ggacgccaat ctcttaaaac 1260 caatctcagt tcggattgta ggctgcaact cgcctacat aagtcggaat cgctagtaat 1320 cgcggatcag catgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac 1380 cacgggagtt tgcaacaccc gaagtcggtg aggtaaccgc aaggagccag ccgccgaagg 1440 tggggtagat gactggggtg aagtcgta 1468

Claims (7)

delete Brevibacillus brevis HK544 strain showing plant disease control activity deposited with accession number KACC 81093BP, the culture solution of the strain, and the culture filtrate of the strain. Any one or more selected from the group consisting of an active ingredient, The plant disease is a composition for controlling plant diseases, characterized in that the cabbage black pattern disease, red pepper anthracnose disease, Phalaenopsis bacterial brown spot disease, bacterial hole disease, kiwi ulcer disease and tomato late blight.
3. The method of claim 2,
The cabbage black pattern disease is Alternaria brassicicola (Alternaria brassicicola), pepper anthrax is Colletotrichum coccodes (Colletotrichum coccodes), Phalaenopsis Bacterial brown spot pattern disease Exidoborax avene subsp. Cattle ray (Acidovorax avenae subsp. cattleyae), bacterial pore disease is Pv. Pruni (Xanthomonas arboricola pv. pruni), kiwi ulcer disease Pseudomonas syringe pv. Actini die (Pseudomonas syringae pv. actinidiae) and tomato blight is phytophthora infestans (Phytophthora infestans) is the causative agent of a composition for controlling plant diseases.
A method for controlling a plant disease comprising the step of treating the composition for controlling a plant disease of claim 2 in a plant or the surrounding environment of the plant.
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