Composition for enhancing plant disease resistance containing a Bacillus amyloliquefaciens strain and use thereof

Abstract

The present application relates to a composition for enhancing resistance to plant diseases, a composition for controlling plant diseases, which comprises the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P and / or a culture thereof, a method for enhancing resistance to plant diseases, a method for preventing or treating plant diseases, and a method for controlling plant diseases, using the same, the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P, use of the microorganism or composition for enhancing resistance to plant diseases, use for controlling plant diseases, and use for preventing or treating plant diseases.

Description

[Technical field]

[0001] The present application relates to a composition for enhancing resistance to plant diseases, a composition for controlling plant diseases, which comprises the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P and / or a culture thereof, a method for enhancing resistance to plant diseases, a method for preventing or treating plant diseases, and a method for controlling plant diseases, using the same, the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P, use of the microorganism or composition for enhancing resistance to plant diseases, use for controlling plant diseases, and use for preventing or treating plant diseases. [Background technology]

[0002] Chemical pesticides and fertilizers have contributed greatly to improving agricultural productivity and freeing mankind from hunger. Some compounds have been used in agricultural and horticultural fungicides and insecticides (Patent Document 1), but recently, the negative aspects of toxicity to humans and animals and environmental pollution have attracted more attention than the positive effects, and there has been growing interest in environmentally friendly pest control farming methods that reduce or eliminate the use of synthetic pesticides to cultivate crops.

[0003] In particular, biological control using microorganisms or plant extracts for controlling agricultural pests has attracted attention, as such biological control can replace chemical pesticides that are harmful to the environment, thereby providing safe agricultural products to consumers. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] U.S. Patent No. 10,123,537 Summary of the Invention [Problem to be solved by the invention]

[0005] The problem to be solved by the present application is to provide a composition for enhancing resistance to plant diseases, a composition for controlling plant diseases, which comprises the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P, a method for enhancing resistance to plant diseases, a method for preventing or treating plant diseases, and a method for controlling plant diseases, using the same, the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P, use of the microorganism or composition for enhancing resistance to plant diseases, use for controlling plant diseases, and use for preventing or treating plant diseases. [Means for solving the problem]

[0006] The present application aims to provide a composition for enhancing resistance to plant diseases and a composition for controlling plant diseases, which contain the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and / or a culture thereof.

[0007] Another object of the present application is to provide a composition for enhancing resistance to plant diseases and a composition for controlling plant diseases, which contain the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and valine.

[0008] A further object of the present application is to provide a method for enhancing plant disease resistance using the composition for enhancing plant disease resistance.

[0009] A further object of the present application is to provide a method for preventing or treating a plant disease using the composition for enhancing plant disease resistance.

[0010] A further object of the present application is to provide a method for controlling plant diseases using the composition for controlling plant diseases.

[0011] The present application further aims to provide a Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P.

[0012] Furthermore, the present application aims to provide uses of the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P for enhancing plant disease resistance, for controlling plant diseases, and for preventing or treating plant diseases.

[0013] Furthermore, the present application aims to provide uses of a composition comprising the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P and valine for enhancing plant disease resistance, for controlling plant diseases, and for preventing or treating plant diseases. Effect of the Invention

[0014] The Bacillus amyloliquefaciens N351 strain deposited under the accession number KCCM13138P has the effect of enhancing the disease resistance of the plant body, and the use of a mixture of the strain and valine can enhance the disease resistance of the plant body more effectively. [Brief description of the drawings]

[0015] [Figure 1] FIG. 13 is a diagram showing that the Bacillus amyloliquefaciens N351 strain has superior control effects compared to other Bacillus strains and product controls, based on leaf photographs and measurement results of the length of disease symptoms. [Diagram 2] FIG. 1 shows the bactericidal effect (plant disease control effect) of Bacillus amyloliquefaciens N351 strain against various plant pathogens, based on photographs of disks with confirmed inhibition zones and measurements of the diameters of the inhibition zones. [Diagram 3] FIG. 1 shows the biofilm formation effect of Bacillus amyloliquefaciens strain N351. [Figure 4] This figure shows the excellent pest control effect of Bacillus amyloliquefaciens strain N351 based on leaf photographs taken with a hyperspectral camera and a visible light camera, as well as the NDVI index. [Diagram 5]FIG. 1 is a graph showing the excellent control effect of Bacillus amyloliquefaciens N351 strain against soybean stem rot, cucumber bacterial spot, and lettuce soft rot in terms of disease severity and control value. [Figure 6] This figure shows the excellent control effect of a powder formulation of Bacillus amyloliquefaciens N351 strain against rice blast, tomato gray mold, wheat leaf rust, barley powdery mildew, and pepper anthracnose by pot assay. [Figure 7] FIG. 1 shows the control effects of a powder formulation of Bacillus amyloliquefaciens N351 strain and a mixture with valine against soybean stem rot and cucumber bacterial spot by pot assay. [Figure 8] FIG. 1 shows the pest control effects of a powder formulation of Bacillus amyloliquefaciens N351 strain and a valine mixture in a glasshouse test. [Figure 9] FIG. 1 shows the ability of Bacillus amyloliquefaciens N351 strain powder preparation and valine mixture to induce induced resistance, based on pathogen count and GUS activity. [Figure 10] FIG. 1 shows the plant growth-promoting ability of Bacillus amyloliquefaciens N351 strain powder formulation and valine mixture in soybean and cucumber. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] These will be described in detail below. Each description and embodiment disclosed in this application is also applicable to other descriptions and embodiments. That is, all combinations of various elements disclosed in this application are included in this application. In addition, this application is not limited to the following specific description. Furthermore, many papers and patent documents are referenced throughout this specification, and citations are provided. The disclosure contents of the cited papers and patent documents are incorporated in their entirety into this specification by reference, thereby more clearly explaining the state of the art to which this application pertains and the contents of this application.

[0017] One aspect of the present application provides a composition for enhancing resistance to plant diseases, comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

[0018] In this application, the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P is designated N351 CP02-1001, which is synonymous with and may be used interchangeably with Bacillus amyloliquefaciens N351 microorganism, Bacillus amyloliquefaciens N351 strain, N351 microorganism, N351 strain, Bacillus amyloliquefaciens N351 CP02-1001 microorganism, Bacillus amyloliquefaciens N351 CP02-1001 strain, N351 CP02-1001 microorganism and N351 CP02-1001 strain.

[0019] In one embodiment, the N351 microorganism may have the ability to enhance plant disease resistance.

[0020] In one embodiment, the N351 microorganism may have the ability to control plant diseases.

[0021] In one embodiment, the N351 microorganism may have the ability to prevent or treat a plant disease.

[0022] In one embodiment, the N351 microorganism may form a biofilm.

[0023] In one embodiment, the N351 microorganism has a superior plant disease resistance enhancing ability, plant disease control ability and / or biofilm forming ability to control Bacillus amyloliquefaciens, Bacillus velezenesis and Bacillus siamensis, which are taxonomically classified in the Bacillus amyloliquefaciens group, and Bacillus subtilis, but is not limited thereto. The control strain may be Bacillus amyloliquefaciens KACC10116, Bacillus velezensis QST713 and Bacillus siamensis KACC15859, and the Bacillus subtilis may be Bacillus subtilis KACC10854PB3.

[0024] In one embodiment, the N351 microorganism is used for enhancing plant disease resistance, controlling plant diseases, and / or preventing or treating plant diseases in at least one selected from the group consisting of, but not limited to, beans, fruit vegetables, leafy vegetables, root vegetables, and cereals.

[0025] In one embodiment, the N351 microorganism may be Phytophthora sojae, Pseudomonas syringae pv. lachrimans, Pectobacterium carotovorum subsp. Carotovorum, Magnaporthe oryzae, Botrytis cinerea, Puccinia recondite, Blumeria graminis f.sp. hordei, Colletotrichum coccodes, Pseudomonas syringae pv. tomato, Rhizoctonia solani, Pythium ultimum, Colletotrichum truncatum, Colletotrichum acutatum, Sclerotinia sclerotiorum, Phytophthora capsici, Phytophthora infestans, Fusarium oxysporum, and Pseudomonas syringae pv. tabaci, but are not limited thereto.

[0026] By way of example, and not limitation, the N351 microorganism produces a variety of secondary metabolites, is heat stable, and forms endospores that are resistant to adverse environments.

[0027] The culture product in this application includes a medium obtained by culturing a specific microorganism in a culture medium, a culture solution, an extract, a dilution, a concentrate, a filtrate, a dried product, a lysate obtained by collecting and crushing the cells of the microorganism, a fermented product, a purified product, and / or any form of substance that can be formed using them, and the culture solution means one that contains the specific microorganism, and the culture filtrate means one that does not substantially contain the specific microorganism (here, substantially means that the specific microorganism separated by filtration or the like is excluded, and does not mean that the microorganism is completely excluded from the filtrate).

[0028] In the present application, "resistance to plant diseases" refers to a property of preventing the invasion of pathogens, a property of preventing the onset of plant diseases, and / or a property of not becoming ill even if a pathogen invades. The resistance to plant diseases includes, but is not limited to, induced resistance. Plants have an immune system that resists various attacks by pathogens, and this immune system causes disease resistance reactions to appear not only in leaves invaded by pathogens, but also in the entire plant body not invaded by pathogens. Such an immune system is called induced resistance, and by using this, it is possible to overcome the shortcomings of biological control agents and maximize their efficacy.

[0029] As an example, the composition for enhancing plant disease resistance of the present application induces induced systemic resistance in a plant, but is not limited thereto.

[0030] As an example, the composition for enhancing plant disease resistance of the present application enhances plant disease resistance without inhibiting plant growth, but is not limited thereto. Specifically, derivatives that induce various resistance reactions, not just to pathogens, are known as substances that induce plant resistance, and some of them (e.g., benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester, benzothiadiazole; BTH) cause serious side effects such as suppressing plant growth and significantly reducing yield. However, the composition for enhancing plant disease resistance of the present application does not suppress plant growth, but rather promotes plant growth.

[0031] As an example, the composition for enhancing plant disease resistance of the present application may be one that prevents or treats a plant disease caused by infection of a plant with a pathogen, or one that induces induced systemic resistance in a plant, but is not limited thereto.

[0032] In the present application, "prevention" refers to any action that suppresses or delays infection by pathogens, the onset of plant diseases, or the progression thereof.

[0033] In this application, "treatment" refers to any action that improves or beneficially alters the symptoms of a plant disease. In this application, treatment includes any action that prevents infection by a pathogen or recurrence of a plant disease, alleviates symptoms, reduces the rate of progression, alleviates the disease state, temporarily or permanently alleviates the disease state, brings about recovery, or improves the prognosis.

[0034] In this application, the term "plant disease" refers to a disease that occurs in plants, and means that abnormalities occur in the original morphology or physiological functions of the plant. The cause of the disease is a disease caused by a pathogen, a nutrient deficiency in the soil, weather conditions, toxic gases, etc. The pathogen is a fungal pathogen or a bacterial pathogen, but is not limited thereto.

[0035] As an example, the plant disease of the present application is at least one selected from the group consisting of Phytophthora stem and root rot, bacterial leaf blight, bacterial soft rot, rice blast, gray mold disease, leaf rust, powdery mildew, anthracnose, bacterial speck, damping off, Sclerotinia rot, Fusarium wilt, and wild fire, but is not limited thereto.

[0036] In this application, "Phytophthora stem and root rot" is a type of mold disease that occurs in potatoes, tomatoes, etc., and the external symptoms vary slightly depending on the level of infection. During the vigorous growth period, the plant body shrinks, the lower leaves take on a reddish-brown color, and the young leaves fade and take on various colors. When the infection is not severe, the external symptoms are often not obvious, and the fruits of the diseased plants are small and the stolon formation is poor. The stem and root rot mainly occurs in potatoes, beans, and fruit vegetables, but can occur in any type of plant. The stem rot is caused by, but is not limited to, pathogenic fungi of the genus Phytophthora belonging to the family Pythiaceae of flagellate fungi, specifically Phytophthora sojae, Phytophthora capsici, and / or Phytophthora infestans.

[0037] In one embodiment, the stem rot disease is, but is not limited to, soybean stem rot disease, pepper stem rot disease, or potato stem rot disease.

[0038] In this application, "bacterial leaf blight" refers to a disease in which spots appear and turn brown. The spots are formed on leaves, stems, pods, petioles, cotyledons, etc. The disease occurs on any type of plant, including soybean, mung bean, and fruit vegetables. The bacterial leaf blight is induced by a microorganism of the genus Pseudomonas, specifically, Pseudomonas syringae pv. lachrymans, but is not limited thereto.

[0039] In one embodiment, the bacterial spot disease is, but is not limited to, cucumber bacterial spot disease.

[0040] In this application, "bacterial soft rot" refers to a disease in which the tissues of a plant are eroded, causing a foul odor, rotting, and soreness. It is particularly prevalent in vegetables with high moisture content, such as lettuce and Chinese cabbage, but can also occur in tomatoes, sweet potatoes, potatoes, and other plants, and can occur in any type of plant. The soft rot is induced by necrotic parasitic bacteria, specifically, but not limited to, Pectobacterium carotovorum subsp. carotovorum.

[0041] In one embodiment, the soft rot disease is lettuce soft rot disease, but is not limited thereto.

[0042] In the present application, "rice blast" is the most important disease of cereals, especially rice, and occurs in all parts of cereals. Lesions form on aboveground parts such as leaves, panicles, rachises, nodes, and rice grains, with blast occurring most frequently on leaves, panicles, and rachises. Infection occurs by conidia, and the general symptoms are that irregular red, gray, black, and / or brown spots spread on the leaves, and the entire leaf turns brown and withers. The head of the panicle and rachises wither and turn light brown, and gray mold develops on the surface when there is a lot of moisture. Dark brown lesions form on the nodes, which become easily broken, and in humid environments the surface is covered with gray mold. The rice blast is induced by, but is not limited to, the genus Magnaporthe microorganism, specifically Magnaporthe oryzae.

[0043] In one embodiment, the blast disease is rice blast disease, but is not limited thereto.

[0044] In this application, "gray mold disease" refers to a disease in which mold fungi invade plants and cause the rotting of leaves, flowers, fruits, branches, etc. The fungi live in the form of mycelium or sclerotium in the soil or in the remains of leaves, flowers, fruits, branches, etc., and then cause the petals and fruits to rot dark brown and form gray-green mycelium and spores. As the disease progresses, fruit drop and branch dieback also occur. Gray mold disease occurs mainly in citrus and tomatoes, but can occur in any type of plant. The gray mold disease is induced by Botrytis genus microorganisms, specifically Botrytis cinerea, but is not limited thereto.

[0045] In one embodiment, the gray mold is tomato gray mold, but is not limited thereto.

[0046] In this application, "leaf rust" refers to a disease in which raised lesions are formed on plants. It occurs not only on leaves but also on other parts such as stems, and reddish brown powder appears on the raised lesions. Although there is a slight difference in color from the lesions, the lesions grow larger, merge, turn yellow, and as the disease progresses, the plant dies. It often occurs when winters are warm and precipitation is abundant, resulting in good growth and luxuriance in spring crops, and the occurrence of the disease also increases when sowing is early and a lot of nitrogen fertilizer is used. The leaf rust disease mainly occurs on wheat, but can occur on any type of plant. The leaf rust disease is induced by microorganisms of the genus Puccinia, specifically Puccinia hordei or Puccinia recondite, but is not limited thereto.

[0047] In one embodiment, the leaf rust disease is wheat leaf rust disease, but is not limited thereto.

[0048] In this application, "powdery mildew" refers to a disease in which a white or grayish-white mold covers the surface of a plant body. The lesions spread from the lower leaves to the leaf sheaths, stems, and rachises, and small black spots appear on the grayish-white lesions in the later stages. The disease occurs more frequently in years when the cultivation period in spring is warm and rainy, and also occurs when the plant is cultivated in a poorly ventilated place or when fertilizer with a high nitrogen content is used. Powdery mildew mainly occurs on barley, wheat, and other wheat-related crops, but can occur on any type of plant. The powdery mildew is induced by microorganisms of the genus Blumeria, specifically Blumeria graminis f.sp. hordei, but is not limited thereto.

[0049] In one embodiment, the powdery mildew is, but is not limited to, barley powdery mildew.

[0050] In this application, "anthracnose" refers to a disease that mainly causes fruit to rot. It increases under hot and humid conditions such as during the rainy season. Anthracnose can also occur on leaves and stems, but mainly on fruits. Light green spots appear on the fruits, and then ulcer symptoms appear. It mainly occurs on various plants such as fruit vegetables, citrus fruits, and legumes, but can occur on any type of plant. The anthracnose is induced by microorganisms of the genus Colletotrichum, specifically, Colletotrichum coccodes, Colletotrichum truncatum, and / or Colletotrichum acutatum, but is not limited thereto.

[0051] In one embodiment, the anthracnose is, but is not limited to, pepper anthracnose.

[0052] In the present application, the term "leaf spot disease" refers to a disease in which large and small spots (spots or specks) appear on the stems, fruits, and leaves of a plant body. The spots are black, light brown, or brown. The spot disease occurs mainly in fruit vegetables, but can occur in any type of plant. The spot disease is caused by various types of fungi or bacteria, and is induced by, but not limited to, living parasites, specifically microorganisms of the genus Pseudomonas, more specifically Pseudomonas syringae pv. tomato, and even more specifically Pseudomonas syringae pv. tomato DC3000.

[0053] In this application, "damping off" refers to a disease that causes a constricting symptom. It induces constricting symptoms, germination disorders due to seed rot, wilting of young seedlings after germination, withering symptoms, and / or rot of roots and stems. The damping off mainly occurs in fruit vegetables or root vegetables such as chili peppers, tomatoes, eggplants, watermelons, ginseng, beans, cucumbers, and onions, but can occur in any type of plant. The damping off is induced by microorganisms of the genus Rhizoctonia or the genus Pythium, specifically Rhizoctonia solani or Pythium ultimum, but is not limited thereto.

[0054] In this application, "Sclerotinia rot" refers to a disease in which a pathogen spends its time in the plant body and / or soil as sclerotia or hyphae, and then gradually rots the plant. As hyphae grow at the infected site, they soften and rot, forming sclerotia. As the disease progresses, the lesions expand and the entire plant rots. It mainly occurs on fruit vegetables or root vegetables such as cucumber, garlic, and carrot, but can occur on any type of plant. The Sclerotinia rot is induced by a microorganism of the genus Sclerotina, specifically, Sclerotinia sclerotiorum, but is not limited thereto.

[0055] In this application, "Fusarium wilt" refers to a disease that causes wilt symptoms. When the wilt develops, the growth of the plant is inhibited, the leaves turn yellow, and the xylem of the roots turn brown. The wilt mainly develops in fruit vegetables such as strawberries, but can develop in any type of plant. The wilt is induced by a microorganism of the genus Fusarium, specifically Fusarium oxysporum, but is not limited thereto.

[0056] In this application, "wild fire" refers to a disease that causes plants to die as if they were on fire. As the disease progresses, spots appear and the plant becomes necrotic, turns brown or black, and develops yellow lesions on the plant. Wild fire primarily occurs in tobacco and legume plants, but can occur in any type of plant. Wild fire is induced by a microorganism of the genus Pseudomonas, specifically, Pseudomonas syringae pv. tabaci, but is not limited thereto.

[0057] In one embodiment, the plant disease may be caused by Phytophthora sojae, Pseudomonas syringae pv. lachrimans, Pectobacterium carotovorum subsp. Carotovorum, Magnaporthe oryzae, Botrytis cinerea, Puccinia recondite, Blumeria graminis f.sp. hordei, Colletotrichum coccodes, Pseudomonas syringae pv. tomato, Pseudomonas syringae pv. tomato, Rhizoctonia solani, Pythium ultimum, Colletotrichum truncatum, Colletotrichum acutatum, Sclerotinia sclerotiorum, Phytophthora capsici, Phytophthora infestans, Fusarium oxysporum, and Pseudomonas syringae pv. tabaci, but is not limited thereto.

[0058] In one embodiment, the plant disease may be phytophthora sojae, phytophthora capsici, and / or phytophthora infestans, stem rot caused by Pseudomonas syringae pv. lachrymans, soft rot caused by Pectobacterium carotovorum subsp. carotovorum, rice blast caused by Magnaporthe oryzae, gray mold caused by Botrytis cinerea, Puccinia recondita, and the like. recondite, powdery mildew caused by Blumeria graminis f.sp. hordei, anthracnose caused by Colletotrichum coccodes, Colletotrichum truncatum and / or Colletotrichum acutatum, leaf spot caused by Pseudomonas syringae pv. tomato, damping off caused by Rhizoctonia solani and / or Pythium ultimum, Sclerotinia sclerotiorum sclerotiorum, Fusarium wilt caused by Fusarium oxysporum, and wildfire caused by Pseudomonas syringae pv. tabaci, but is not limited thereto.

[0059] The plant disease may occur in any type of plant, but in one embodiment, the plant may be selected from the group consisting of beans, fruit vegetables, leafy vegetables, root vegetables, and grains.

[0060] In the present application, "legumes" refers to legumes (Fabaceae) that are used as food or feed. Examples include soybean, wild bean, kidney bean, black bean, pea, adzuki bean, peanut, cowpea, etc. Specifically, the legume is soybean, but is not limited thereto.

[0061] In the present application, "fruit vegetables" refers to vegetables intended for use as fruits when classified according to the part of the vegetable that is used. Examples include chili peppers, cucumbers, tomatoes, watermelons, strawberries, and Japanese cantaloupe. Specifically, the fruit vegetables are chili peppers, cucumbers, and tomatoes, but are not limited thereto.

[0062] In the present application, "leafy vegetables" refers to vegetables intended for use as leaves when classified according to the part of the vegetable that is used. Examples include Chinese cabbage, lettuce, perilla, spinach, etc. Specifically, the leafy vegetables are Chinese cabbage and lettuce, but are not limited thereto.

[0063] In the present application, the term "root vegetables" refers to vegetables that are intended to be used for their roots when classified according to the part of the vegetable that is used. Examples of root vegetables include, but are not limited to, garlic and ginseng.

[0064] In the present application, "cereals" refers to cultivated plants of the Gramineae family whose seeds are used for food. Examples include rice, wheat, barley, corn, etc. Specifically, the cereals are rice, wheat, or barley, but are not limited thereto.

[0065] In one embodiment, the composition for enhancing plant disease resistance may further include valine.

[0066] Valine induces the plant immune system and improves induced resistance. In the present application, the composition for enhancing plant disease resistance containing the N351 microorganism further contains valine, thereby further enhancing plant disease resistance and promoting induced resistance and growth, but is not limited thereto.

[0067] As an example, the valine of the present application is contained in an amount of 0.0001 to 10%, 0.0001 to 5%, 0.0001 to 1%, 0.005 to 10%, 0.005 to 1%, 0.005 to 0.5%, 0.001 to 10%, 0.001 to 1%, 0.001 to 0.5%, 0.01 to 10%, 0.01 to 1%, 0.01 to 0.5%, 0.05 to 10%, 0.05 to 1%, 0.05 to 0.5%, 0.1 to 10%, 0.1 to 1%, or 0.1 to 0.5% (w / w) relative to the entire composition for enhancing plant disease resistance or the entire composition for controlling plant disease, but the amount is not limited thereto.

[0068] The composition for enhancing resistance to plant diseases of the present application may be any formulation or product necessary for enhancing resistance to plant diseases, such as an agricultural chemical formulation, a fertilizer, or a control agent.

[0069] In this application, the term "pesticides" refers to chemicals used for the protection and growth of agricultural crops. Specifically, this term includes not only preparations used to control diseases, pests, weeds, pathogens, nematodes, mites, and the like that occur during the cultivation and storage of agricultural crops, but also growth regulators used to enhance or suppress the physiological functions of agricultural crops and adjuvants that enhance the efficacy of drugs.

[0070] The term "fertilizer" in this application means a nutrient material that enriches the soil and promotes the growth of plants. The fertilizer containing the composition of this application can be easily used by mixing it with the soil before sowing, and can be applied together with the application of pesticides and the like.

[0071] The composition for enhancing plant disease resistance of the present application may further contain substances that are commonly used for enhancing disease resistance, and may further contain, in addition to the N351 microorganism and valine, a pharma- ceutically acceptable solid carrier, liquid carrier, liquid diluent, liquefied gaseous diluent, solid diluent or other auxiliary agent as an excipient, such as a surfactant such as an emulsifier, dispersant, or foaming agent.

[0072] The composition for enhancing plant disease resistance of the present application can be formulated into an agricultural composition by mixing the active ingredient with the excipient, and the method for formulating the agricultural composition may be any method commonly used in the art.

[0073] In addition, the composition for enhancing plant disease resistance of the present application may be provided in the form of a wettable powder, granule, dust, emulsion, spray, smoke, capsule, or gel, or may be provided in a contact agent in a doughnut-shaped form for buoyancy of the composition, but is not limited thereto.

[0074] Another aspect of the present application provides a composition for controlling plant diseases, comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

[0075] The Bacillus amyloliquefaciens microorganism, culture, plant disease, and the like deposited under the accession number KCCM13138P are as described above.

[0076] In one embodiment, the composition for controlling a plant disease may further contain valine.

[0077] In the present application, "control" means preventing damage to plants such as crops by pests, removing pathogens that have appeared, and preventing the spread of pathogens. The control is a series of actions for reducing damage caused by pests, and includes, but is not limited to, physical (mechanical) control methods, cultural control methods, chemical control methods, biological control methods, and environmentally friendly control methods. The control includes not only chemical control using fungicides or insecticides that are toxic to pests, but also methods of creating an environment that pathogens do not like by cultivating resistant plants, crop rotation, etc., methods of removing pathogens by heat treatment of plants or steam treatment of soil, and methods of attracting and killing pests using attractants or sex pheromones.

[0078] The composition for controlling plant diseases of the present application may be any formulation or product necessary for controlling plant diseases, such as an agricultural chemical formulation, a fertilizer, or a control agent.

[0079] The composition for controlling plant diseases of the present application may further contain substances commonly used for control, and may further contain, in addition to the Bacillus amyloliquefaciens microorganism deposited under Accession No. KCCM13138P and valine, a pharma- ceutically acceptable solid carrier, liquid carrier, liquid diluent, liquefied gaseous diluent, solid diluent or other auxiliary agent as an excipient, for example, a surfactant such as an emulsifier, dispersant, or foaming agent.

[0080] The plant disease control composition of the present application can be used by mixing the active ingredient with the excipient and formulating it into an agricultural composition, and the method for formulating it into an agricultural composition may be any method commonly used in the art.

[0081] In addition, the plant disease control composition of the present application may be provided in the form of a wettable powder, granule, dust, emulsion, spray, smoke, capsule, or gel, or may be provided in a contact agent in a doughnut-shaped form to provide buoyancy for the composition.

[0082] Yet another aspect of the present application provides a kit comprising a composition according to the present application (i.e., the composition for enhancing plant disease resistance and / or the composition for controlling plant disease) and valine.

[0083] The composition according to the present application is as described above.

[0084] The kit is intended for, but is not limited to, the control, prevention, or treatment of plant diseases. The kit is provided so that the composition and valine are stored in separate containers and are mixed together when used, but is not limited to this.

[0085] The kit of the present application also includes, but is not limited to, one or more other component compositions, solutions, or devices for preventing or treating plant diseases, enhancing plant disease resistance, or controlling plant diseases. Examples of the other components include, but are not limited to, suitable carriers, solubilizers, buffers, stabilizers, etc. Carriers include soluble carriers and insoluble carriers, and may be, for example, physiologically acceptable buffer solutions known in the art (e.g., PBS), but are not limited to these.

[0086] The kit of the present application may further include a user manual describing optimal reaction conditions. The manual is a printed material that explains how to use the kit, such as reaction conditions. The manual includes information booklets in the form of pamphlets or flyers, labels attached to the kit, and instructions on the surface of the packaging containing the kit. The manual also includes information published or provided via electronic media such as the Internet.

[0087] Yet another aspect of the present application provides a method for enhancing resistance to a plant disease, comprising treating a plant body with a composition for enhancing resistance to a plant disease, the composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

[0088] The Bacillus amyloliquefaciens microorganism deposited under the accession number KCCM13138P, the culture, the plant disease, the plant disease resistance, the composition for enhancing plant disease resistance, and the like are as described above.

[0089] In one embodiment, the composition for enhancing plant disease resistance further includes, but is not limited to, valine.

[0090] In one embodiment, the method for enhancing plant disease resistance also promotes plant growth, but is not limited thereto.

[0091] The plant body may be any part of a plant, such as a root, a stem, a leaf, or a combination thereof, and may be any type of plant. Specifically, the plant body may be, but is not limited to, a plant individual that is at risk of being infected with a plant disease or is infected with a pathogen.

[0092] The treatment with the composition for enhancing plant disease resistance is carried out by spraying, injecting, pouring, mixing, or mixing the composition into soil, etc., of the plant body, but is not limited thereto, as long as the inside or outside of the plant body comes into direct or indirect contact with the composition. The treatment may be carried out simultaneously or at a different time when a pathogen invades the plant, or before or after the pathogen invades the plant, but is not limited thereto.

[0093] In one embodiment, the composition for enhancing plant disease resistance is applied once or twice or more times, but is not limited thereto. When applied twice or more times, the application is performed at intervals of 3 to 15 days, 3 to 10 days, 3 to 7 days, 4 to 7 days, 5 to 7 days, or 6 days, but is not limited thereto.

[0094] In one embodiment, the composition for enhancing plant disease resistance is 5 CFU / ml or more, 1×10 6 CFU / ml or more, 1×10 5 CFU / ml ~ 1 x 10 10CFU / ml, 1×10 5 CFU / ml ~ 1 x 10 9 CFU / ml, 1×10 6 CFU / ml ~ 1 x 10 10 CFU / ml, 1×10 6 CFU / ml ~ 1 x 10 9 CFU / ml, or 1 x 10 3 CFU / ml ~ 1 x 10 12 The treatment is carried out at a concentration of, but not limited to, CFU / ml.

[0095] As one embodiment, the composition for enhancing plant disease resistance is applied to a plant body in an amount of 1 ml to 50 ml, 2 ml to 50 ml, 3 ml to 50 ml, 4 ml to 50 ml, 5 ml to 50 ml, 1 ml to 30 ml, 2 ml to 30 ml, 3 ml to 30 ml, 4 ml to 30 ml, 5 ml to 30 ml, 1 ml to 20 ml, 2 ml to 20 ml, 3 ml to 20 ml, 4 ml to 20 ml, 5 ml to 20 ml, 1 ml to 10 ml, 2 ml to 10 ml, 3 ml to 10 ml, 4 ml to 10 ml, 5 ml to 10 ml, or 5 ml, but is not limited thereto.

[0096] In one embodiment, the plant is at least one selected from the group consisting of beans, fruit vegetables, leafy vegetables, and grains, but is not limited thereto.

[0097] The beans, fruit vegetables, leafy vegetables, grains, etc. are as described above.

[0098] Yet another aspect of the present application provides a method for preventing or treating a plant disease, the method comprising treating a plant body with a composition for enhancing resistance to plant disease, the composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

[0099] The Bacillus amyloliquefaciens microorganism deposited under the accession number KCCM13138P, the plant disease, the plant disease resistance, the composition for enhancing resistance to plant diseases, the treatment with the composition for enhancing resistance to plant diseases, the plant body, prevention, treatment, etc. are as described above.

[0100] Yet another aspect of the present application provides a method for controlling a plant disease, comprising treating a plant body with a composition for controlling a plant disease, the composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under Accession No. KCCM13138P and a culture thereof. Specifically, the method for controlling a plant disease may also promote plant growth.

[0101] The Bacillus amyloliquefaciens microorganism deposited under the accession number KCCM13138P, the culture, the plant, the plant body, the plant disease, the control, the composition for controlling a plant disease, the treatment with the composition for controlling a plant disease, and the like are as described above.

[0102] The treatment with the composition for controlling plant diseases is carried out by spraying, injecting, pouring, mixing, or mixing the composition into soil, etc., of the plant body, as long as the inside or outside of the plant body comes into contact with the composition, but is not limited to these. The treatment may be carried out simultaneously or at a different time when the pathogen invades the plant, or before or after the pathogen invades the plant, but is not limited to these.

[0103] In one embodiment, the composition for controlling plant diseases is applied once or twice or more times, but is not limited thereto. When the composition is applied twice or more times, the application is performed at intervals of 3 to 15 days, 3 to 10 days, 3 to 7 days, 4 to 7 days, 5 to 7 days, or 6 days, but is not limited thereto.

[0104] In one embodiment, the plant protection composition is administered to 1×10 5 CFU / ml or more, 1×10 6 CFU / ml or more, 1×10 5 CFU / ml ~ 1 x 10 10 CFU / ml, 1×10 5 CFU / ml ~ 1 x 10 9 CFU / ml, 1×10 6 CFU / ml ~ 1 x 10 10 CFU / ml, 1×10 6CFU / ml ~ 1 x 10 9 CFU / ml, or 1 x 10 3 CFU / ml ~ 1 x 10 12 The treatment is carried out at a concentration of, but not limited to, CFU / ml.

[0105] In one embodiment, the plant control composition is applied to a plant body in an amount of 1 ml to 50 ml, 2 ml to 50 ml, 3 ml to 50 ml, 4 ml to 50 ml, 5 ml to 50 ml, 1 ml to 30 ml, 2 ml to 30 ml, 3 ml to 30 ml, 4 ml to 30 ml, 5 ml to 30 ml, 1 ml to 20 ml, 2 ml to 20 ml, 3 ml to 20 ml, 4 ml to 20 ml, 5 ml to 20 ml, 1 ml to 10 ml, 2 ml to 10 ml, 3 ml to 10 ml, 4 ml to 10 ml, 5 ml to 10 ml, or 5 ml, but is not limited thereto.

[0106] Yet another aspect of the present application provides a Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P.

[0107] The Bacillus amyloliquefaciens microorganism deposited under the accession number KCCM13138P is as described above.

[0108] Still other aspects of the present application provide uses of the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P for enhancing plant disease resistance, for controlling plant diseases, and for preventing or treating plant diseases.

[0109] Still other aspects of the present application provide uses of a composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under deposit number KCCM13138P and a culture thereof and valine for enhancing plant disease resistance, for controlling plant diseases, and for preventing or treating plant diseases.

[0110] The Bacillus amyloliquefaciens microorganism, culture, plant disease, plant disease resistance, control, prevention, treatment, etc. deposited under the accession number KCCM13138P are as described above. EXAMPLES

[0111] The present application will be described in more detail below with reference to examples. However, these examples are merely preferred embodiments illustrating the present application, and the present application is not limited thereto. Note that technical matters not described in this specification are fully understood and easily implemented by a skilled engineer in the technical field of the present application or a similar technical field. EXAMPLES

[0112] Verification of the control effect of Bacillus strains against plant diseases To verify the control effect of 16 Bacillus strains, soybeans were used and Phytophthora sojae (Ps), which causes soybean stem rot, was used as the pathogen.

[0113] To grow soybeans, seeds were surface sterilized by immersion in 3% sodium hypochlorite (NaOCl) for 5 min. They were then rinsed five times with sterilized water and then sown in bed soil (Field One, Farm Hannon). Four weeks after sowing, six leaves isolated from soybeans were placed on a square dish (125mmX125mm, SPL) covered with filter paper (110 mm, Watman). A Ps pathogen disk (diameter 5mm) was placed on each leaf, and 16 species of Bacillus spp. strains (N474, N477, N481, N484, N485, N489, N490, N491, N494, N495, N311, N339, N343, N351, N352, and N429) were inoculated at 1 × 10 8 500 μl was sprayed onto each leaf at a concentration of cfu / ml.

[0114] The 16 Bacillus strains were cultured in Luria-Bertani broth (LB) solid medium at 30°C for 24 hours. The Ps pathogen was cultured in V8 juice medium (V8-juice, 200 mL; CaCO3, 1 g; agar, 17 g / L) for 7 days. Seven days after treatment with the Bacillus strains, the length of the symptoms on each of the six leaves was measured, and those with statistically different symptoms compared to the product control group, Serenade (Bayer), were selected.

[0115] As shown in Figure 1, when the control effects of the untreated control group and the product control group were compared with the 16 strain-treated groups, the N351 strain-treated group showed the best control effect compared with the other 15 strain-treated groups. Figure 1(a) is a photograph of a leaf, and Figure 1(b) shows the results of measuring the length of the disease symptoms.

[0116] In the untreated group, product control group, and Bacillus-treated group, symptoms of browning caused by Ps appeared on most of the six leaves, but in the N351-treated group, no symptoms appeared even seven days after Ps inoculation. The average length of symptoms in the Serenade-treated group (product control group) was 3.9 cm. Four groups, N494, N339, N351, and N429-treated groups, had shorter symptoms than this, but of these, only the N351-treated group showed statistical significance compared to the Serenade-treated group (product control group).

[0117] As mentioned above, the new strain with the best control effect was named Bacillus amyloliquefaciens N351 and was internationally deposited at the Korea Microorganism Collection, an international depository institution under the Budapest Treaty, on March 2, 2022 under deposit number KCCM13138P. EXAMPLES

[0118] Confirmation of the bactericidal effect (plant disease control effect) of Bacillus amyloliquefaciens N351 strain against various plant pathogens Four paper discs were placed on 8.5 cm diameter Petri dishes containing potato dextrose agar (PDA), V8 juice, Luria-Bertani broth (LB), and King's B (KB) medium, respectively, and the standard strain Bacillus siamensis (B. siamensis T : B. siamensis KACC15859; B.si), the standard strain B. amyloliquefaciens T : B. amyloliquefaciens KACC 10116; Ba), Bacillus subtilis (B. subtilis T Three types of strains were inoculated: B. subtilis KACC 10854PB3 (Bs), Serenade (identified as B. velezensis QST713), and N351. The remaining paper disk was used as an untreated control.

[0119] For fungal pathogens, a colony of the pathogen was placed in the center of a petri dish using a cork borer with a diameter of 5 mm.

[0120] Bacterial pathogens (OD 600 In the case of the OD = 1), 1000 μl of each was spread onto a petri dish, and then the strain to be examined for bactericidal effect was inoculated onto the paper disk. The strain to be examined was cultured in Luria-Bertani broth (LB) solid medium at 30°C for 24 hours, and the strain (OD 600 =1) 50 μl was inoculated onto a paper disc.

[0121] Plant pathogens Rhizoctonia solani (Rs), Pythium ultimum (Pu), Colletotrichum truncatum (Ct), Colletotrichum acutatum (Ca), Sclerotinia sclerotiorum (Ss), Phytophthora sojae (Ps), Phytophthora capsici (Pc), Phytophthora infestans (Pi), Fusarium oxysporum(Fo), Botrytis cinerea(Bc), Pectobacterium carotovorum subsp. Carotovorum(Pcc), Pseudomonas syringae pv. tabaci(Pst) and Pseudomonas syringae pv. lachrimans (Psl), Rs, Pu, Ct, Ca, Ss, Fo and Bc were cultured in PDA medium, Ps, Pc and Pi in V8 juice medium at 28°C for 5 to 7 days before use, Pcc was cultured in LB medium, and Pst and Psl strains were cultured in KB medium at 28°C for 2 days before use.

[0122] Then, each pathogen was cultured in an incubator at 25°C and 20°C for 7 to 20 days, and the diameter of the zone of inhibition was examined.

[0123] As a result, as shown in FIG. 2, the control effect of the N351 strain against various plant pathogens was confirmed. EXAMPLES

[0124] Verification of the biofilm of N351 strain Normally, microorganisms live in communities to survive in their environment, and form biofilms (surface membranes) to do so. The better the biofilm formation, the better the microorganisms will settle in all areas of the plant, so this is used as one of the criteria for selecting useful microorganisms.

[0125] In this example, to measure the level of biofilm formation, a 96-well microplate made of polyvinyl chloride was used, and 100 ul of medium per well was inoculated with microorganisms in an amount that would result in an absorbance of 0.05 at a wavelength of 600 nm, and cultured for 2 days at 28° C. To quantify the amount of biofilm, after culturing the microorganisms, the supernatant was removed, and the microplate was exposed to 100 μl of a 1% crystal violet solution for 15 minutes, after which the remaining crystal violet was removed with water, and 100 μl of 95% ethanol was added to elute the crystal violet that was not removed by the biofilm, and then the absorbance was measured at a wavelength of 595 nm using a microplate reader.

[0126] As a result, as shown in FIG. 3, the N351 strain produced more biofilm than any of the four Bacillus type strains evaluated.

[0127] The N351 strain showed an absorbance value six times higher than that of the untreated control. Ba and Bs showed the same absorbance as the untreated control, while Serenade and B.si showed an absorbance of 1.6, and only a small amount of biofilm was formed. The N351 strain formed 2.3, 5, 2.3, and 4 times more biofilm than serenade, Ba, B.si, and Bs, respectively, clearly indicating its difference from representative strains belonging to the Bacillus amyloliquefaciens group (Bacillus amyloliquefaciens, Bacillus siamensis, and Bacillus berezensis) in microbial taxonomy, and once again confirming that it is a strain with high potential for use as a biological control agent. EXAMPLES

[0128] Verification of the control effect of N351 strain and representative Bacillus strains To compare and verify the control effects of representative strains that belong to the same taxonomic group as the N351 strain, Ps pathogen disks were placed on soybean leaves and each strain was sprayed on them in the same manner as in Example 1. The severity of the disease symptoms was then measured using a hyperspectral camera and a visible light camera.

[0129] Seven days after the pathogen treatment, hyperspectral photographs were taken in the wavelength band of 400 to 1000 nm for each Bacillus treatment group (Figure 4a). Here, normal leaves and diseased leaves have different wavelength bands, which can be displayed by color. Even when it is difficult to distinguish the difference in disease severity with the naked eye, it can be easily distinguished using a hyperspectral camera. The Normalized Difference Vegetation Index (NDVI) was used to quantify the degree of diseased leaves in the total leaf area, and the number of pixels was used to indicate the degree. Generally, a value of 0.7 or higher is considered to be a healthy leaf.

[0130] As a result, even in the visible light range, the untreated control group was observed to have a lighter yellow-green color and faster progression of the disease compared to all of the strain-treated groups (Figure 4a, RGB photo).When photographed with a hyperspectral camera, it was observed that the leaves were photographed in different colors depending on the severity of the disease symptoms, ranging from normal leaves (green) to dead leaves (brown) (Figure 4a, NDVI photo).

[0131] The percentage of healthy leaves in the N351-treated group was 91%, which was 56% higher than the 35% in the untreated control group. In contrast, the percentage of healthy leaves in the other strain-treated groups was only 52% for serenade, 56% for Ba, 73% for B.si, and 56% for Bs. The hyperspectral camera also showed that N351 had excellent soybean stem rot control effects (Figure 4b). EXAMPLES

[0132] Pot test of the control effect of N351 strain The plant disease control effect of the N351 strain was examined on three kinds of crops grown in pots (square pot. 5x5 cm).

[0133] Each crop was grown in a pot in a plant incubator (12 hours of light incubation, 28°C, vision) for 3 weeks, and then each Bacillus strain (OD 600 5 ml of the above mixture (=1) was sprayed onto each plant. The Bacillus strain was cultured in LB solid medium at 30°C for 24 hours before use. Seven days after the first spraying, a second spraying was performed in the same manner as the first, and one day later, the pathogen was inoculated onto each crop.

[0134] The target diseases were stem rot (Phytophthora sojae, Ps) for soybeans, bacterial spot (Pseudomonas syringae pv. lachrymans, Psl) for cucumbers, and soft rot (Pectobacterium carotovorum pv. carotovora, Pcc) for lettuce.

[0135] The Ps strain, which is the causative agent of soybean stem rot, was inoculated into V8 medium (V8 juice, 1.8% agar) and cultured in an incubator at 25°C for 2 weeks. The formed spores were harvested in sterilized distilled water and the spore suspension of a given concentration (5.0 × 10 5 Soybeans inoculated with the spore suspension were allowed to develop the disease for 3 days in a temperature- and humidity-controlled room at 26°C, and the disease severity was assessed on a scale of 0 to 5 according to the following criteria 7 days after the pathogen treatment. 0 = no symptoms, 1 = one leaf withers, 2 = two leaves withers, 3 = three leaves withers, 4 = four leaves withers, 5 = entire plant withers

[0136] The Psl strain, which is the causative agent of bacterial spot disease of cucumber, was cultured on KB (King's B) solid medium at 28°C for 48 hours. The causative agent was cultured at an OD of 1 (10 9 The pathogen-inoculated cucumbers were allowed to develop the disease for three days in a temperature- and humidity-controlled room at 26°C, and the severity of the disease was assessed on a scale of 0 to 5 according to the following criteria seven days after the pathogen treatment. 0=no symptoms, 1=slight disease symptoms, 2=30% disease symptoms, 3=disease and small areas of necrosis, 4=necrosis, 5=large areas of necrosis and wilting symptoms.

[0137] The Pcc strain, which is the pathogen of bacterial soft rot of lettuce, was cultured in LB (Luria-Bertani broth) solid medium at 28°C for 48 hours. The pathogen reached an OD of 1 (10 9 The lettuce plants inoculated with the pathogen were allowed to develop a disease in a temperature- and humidity-controlled room at 26°C for one day, and the severity of the disease was observed 48 hours after the pathogen treatment, and the severity of the disease was rated on a scale of 0 to 5 according to the following criteria. 0 = no symptoms, 1 = mild soft rot symptoms at the inoculation site, 2 = soft rot symptoms at the inoculation site, 3 = severe soft rot symptoms at the inoculation site, 4 = soft rot symptoms around the inoculation site, 5 = soft rot symptoms on the entire leaf

[0138] For soybean stem rot, cucumber bacterial spot and lettuce soft rot, the control effect (control value) of the N351 strain-treated group compared to the untreated control group was calculated from the disease severity as follows.

[0139]

number

[0140] In soybean stem rot, the N351-treated group had a disease severity of 1.5, compared to 3.5 in the water control group. The control value was 57% of the untreated control group. The N351-treated group also showed a better disease control effect than the serenade-treated group, which had a control value of 23% (Figure 5a).

[0141] In the case of bacterial spot disease on cucumbers, the disease severity in the N351-treated group was 1.6, which was reduced from 3.3 in the water control group. The control value was 52% of the untreated control group. The N351-treated group showed a better disease control effect than the serenade-treated group, which had a control value of 18% (Figure 5b).

[0142] In the case of lettuce soft rot, the N351-treated group had a disease severity of 1.3, a significant reduction compared to the water control group, which had a disease severity of 4. The control value was 68% of the untreated control group. The N351-treated group showed excellent disease control effect, similar to that of the serenade-treated group, which had a control value of 60% (Figure 5c). EXAMPLES

[0143] Pot test of the control effect of powder formulation of N351 strain In order to observe whether the N351 strain alone or in powder form would have a superior control effect against plant diseases, the N351 strain was made into a powder form.

[0144] For this purpose, the N351 strain was cultured in LB solid medium at 28°C for 24 hours, and one colony was inoculated into LB liquid medium. The liquid culture was carried out at 28°C and 200 rpm for 48 hours. The culture solution was centrifuged (4,000 rpm, 20 minutes) to separate the bacteria and the culture filtrate. Diatomaceous earth of the same weight as the bacteria was added, and the mixture was dried by heating at 30°C for 3 days to obtain a powder.

[0145] The prepared N351 powder formulation (1 × 10 9 The 100-fold diluted CFU / g was sprayed on seedlings of each plant that had been prepared and grown in a greenhouse, and one day later, each plant was inoculated with one of five plant pathogens.

[0146] Five pathogens on five different crops were targeted: rice blast (Magnaporthe oryzae), tomato grey mold (Botrytis cinerea), wheat leaf rust (Puccinia recondite), barley powdery mildew (Blumeria graminis f. sp. hordei), and pepper anthracnose (Colletotrichum coccodes).

[0147] For rice blast disease (RCB), the pathogen Magnaporthe oryzae KI-1113a strain was inoculated onto rice bran agar medium and cultured in an incubator at 25°C for 2 weeks. The formed spores were harvested in sterilized distilled water and the spore suspension of a given concentration (5.0 × 10 5 The rice plants inoculated with the spore suspension were left in a moist room in the dark for 24 hours, and then allowed to develop disease in a constant temperature and humidity room at a relative humidity of 80% and a temperature of 25°C for 4 days, and the lesion area rate (%) was examined.

[0148] For tomato gray mold (TGM), the pathogenic fungus Botrytis cinerea was inoculated onto potato agar medium and incubated in an incubator at 20°C (in the dark). The spores formed were used as the inoculum. To inoculate the pathogen, the spores were harvested and the spore concentration was measured using a hemocytometer to determine the concentration of 5.0 × 10 5 The sample was diluted to spores / ml and then sprayed onto tomato seedlings (at the 2-3 leaf stage) that had been treated with the sample. The inoculated tomato seedlings were left in a moist room at 20°C (relative humidity 95% or higher) for three days to induce disease, and the lesion area rate (%) was then investigated.

[0149] In the case of wheat leaf rust (WLR), the pathogen Puccinia recondita is a living parasite, so spores (urediniospores) formed on wheat leaves obtained by directly subculturing plants in the laboratory were used as the inoculum. For the efficacy study, five wheat seeds (Keumgang wheat) were sown in disposable pots (diameter: 4.5 cm) and the one-leaf stage seedlings grown in a greenhouse for eight days were treated with the sample, and one day later, the inoculum (0.67 g / L spores) was sprayed for inoculation. The inoculated wheat seedlings were treated in a moist room at 20°C for one day, and then transferred to a constant temperature and humidity room at 20°C with a relative humidity of 70% to induce disease development, and the lesion area rate (%) was investigated seven days after inoculation.

[0150] In the case of barley powdery mildew (BPM), the pathogen Blumeria graminis f. sp. hordei is a parasitic fungus, so barley seedlings were subcultured in the laboratory and the spores formed on the barley leaves were used as the inoculum source. For the efficacy study, five barley seeds ('Hanyoung barley') were sown in disposable pots (diameter: 4.5 cm) and the seedlings at the one-leaf stage were grown in a greenhouse for eight days. The samples were sprayed on the seedlings and air-dried in the greenhouse. The barley treated with the samples was then inoculated by sprinkling powdery mildew spores on them. The inoculated barley seedlings were left in a constant temperature and humidity room at 20°C and a relative humidity of about 50% for seven days to allow the disease to develop, and the lesion area rate (%) was investigated.

[0151] For pepper anthracnose (PAN), the pathogen Colletotrichum coccodes was inoculated into oatmeal medium and cultured at 25°C for 10 days. The spores formed were harvested and the spore concentration was 5 × 10 per ml. 5 A spore suspension was prepared by adjusting the amount of spores so that the spores would form. The prepared spore suspension was sprayed onto pepper seedlings (3-4 leaf stage) treated with the sample, which were then left in a moist chamber (25°C) for 2 days, after which the disease was allowed to develop in a constant temperature and humidity chamber (25°C, 80% RH). Three days after inoculation, the lesion area rate (%) formed on the pepper leaves was investigated. The lesion area rate from the disease investigation and the control value using it were calculated using the following formula.

[0152]

number

[0153]

number

[0154] As a result, as shown in Figure 6, when the control effect of the N351 powder formulation was examined against five types of pathogenic bacteria, it was observed that it had a superior control effect against all five types of diseases compared to the product control group.

[0155] For rice blast disease (Figure 6a), the group treated with the N351 powder formulation showed a higher control value (78%) than the untreated control group (0%) and a control value 13 times higher than that of the product control group (6%).

[0156] For tomato gray mold (Figure 6b), the group treated with the N351 powder formulation showed a higher control value (63%) than the untreated control group (0%) and a control value 3.6 times higher than the product control group (18%).

[0157] In wheat leaf rust (Fig. 6c), the N351 powder formulation treatment group showed a higher control value (67%) than the untreated control group (0%), whereas the product control group had no control effect.

[0158] In barley powdery mildew (Fig. 6d), the N351 powder formulation treatment group showed a higher control value (42%) than the untreated control group (0%), whereas the product control group had no control effect, as in wheat leaf rust.

[0159] For pepper anthracnose (Figure 6e), the group treated with N351 powder formulation showed a higher control value (38%) than the untreated control group (0%) and a five-fold higher control value than the product control group (8%).

[0160] From the above results, it was confirmed that the N351 strain powder formulation showed 3.6 times or more higher control value against five types of diseases compared to the product control group. EXAMPLES

[0161] Pot test of the control effect of N351 powder formulation and valine mixed treatment group Valine was also used to further enhance the plant disease control effect of N351 powder formulation, which has a superior control effect compared to the product control group. N351 powder formulation was mixed with valine to kill pathogens and increase the plant's immunity, maximizing the overall control effect.

[0162] For this purpose, the control effect of N351 powder formulation and valine mixed treatment group was verified for three kinds of plants cultivated in pots (square pot. 5x5 cm). When mixed with N351 powder formulation, valine was used at a concentration of 0.05%. The target was stem rot (Phytophthora sojae) for soybean, and bacterial spot (Pseudomonas syringae pv. lachrymans) for cucumber. Preparation of soybean and cucumber plants, preparation of Bacillus strains, preparation of soybean stem rot and cucumber bacterial spot bacteria, and induction of disease were performed in the same manner as in Example 5. The pathogens were cultured at OD=1 (10 9 The inoculated lettuce was placed in a temperature and humidity controlled room at 26°C for one day to allow the disease to develop, after which the severity of the disease was observed. The control value was calculated as follows:

[0163]

number

[0164] As a result, as shown in Figure 7, when the control effects were investigated for soybean stem rot and cucumber bacterial spot, the N351 powder formulation and valine mixed treatment group were observed to have improved control effects compared to the single treatment group.

[0165] The control value (%) of soybean stem rot was 11% for serenade, 36% for valine, and 51% for the N351 powder formulation group compared to the untreated group, but the control value of the N351 powder formulation and valine mixture treatment group was 64%, which was 13% higher than the N351 powder formulation treatment group. The control value of the N351 powder formulation and valine mixture treatment group was statistically significantly higher than each single treatment group. Compared to the product control group, the N351 powder formulation treatment group had a control value 40% higher, but even more so, the N351 powder formulation and valine mixture treatment group had a control value 53% higher (Figure 7a).

[0166] The control value (%) of cucumber bacterial spot disease was 10% for serenade, 38% for valine, and 60% for the N351 powder formulation treatment group compared to the untreated group, but the control value of the N351 powder formulation and valine mixture treatment group was 69%, which was 10% higher than the N351 powder formulation treatment group. Compared to the product control group, the N351 powder formulation treatment group had a control value 50% higher, but even more so, the N351 powder formulation and valine mixture treatment group had a control value 59% higher (Figure 7b). EXAMPLES

[0167] Glasshouse test of the control efficacy of N351 powder formulation and valine mixture treatments In the pot verification of soybeans and cucumbers in a plant culture vessel, it was observed that the N351 powder formulation and valine mixed treatment group had a superior plant disease control effect compared to the single treatment group, so it was verified whether the disease control effect could be reproduced in a glass greenhouse environment. The subject of the experiment was soybeans. The plant preparation, N351 powder formulation and valine treatment, and soybean stem rot treatment methods were the same as in Example 7. After the stem rot treatment, the disease was allowed to develop for 2 days in a constant temperature and humidity room at 28°C in a glass greenhouse, and the disease severity was then observed. The control value was calculated as follows.

[0168]

number

[0169] As a result, as shown in Figure 8, when the control effect of the N351 powder formulation and valine mixed treatment group against soybean stem rot was investigated, it was observed even in a glass greenhouse that the control effect was improved compared to the single treatment group.

[0170] The control value (%) for soybean stem rot was 7.3% for serenade, 24% for valine, and 44% for the N351 powder formulation treatment group compared to the untreated group, but the control value for the N351 powder formulation and valine mixture treatment group was 76%, indicating that the control value was higher in the mixture treatment than in the single treatment. The control values ​​of the N351 powder formulation and valine mixture treatment groups were statistically significantly higher than those of each single treatment group. EXAMPLES

[0171] Verification of induced resistance in N351 powder formulation and valine-mixed treatment groups The effect of mixing N351 powder formulation with valine on inducing induced resistance was examined.

[0172] For this purpose, Arabidopsis thaliana was used as a model plant, and Pseudomonas syringae pv. tomato DC3000 (hereafter referred to as Pst), which causes spot disease, was used as the pathogen. To grow the plants, Arabidopsis seeds were surface-sterilized by immersing them in 3% sodium hypochlorite (NaOCl) for 5 minutes. They were then rinsed five times with sterilized water and then sown in pots. The plants were grown in a plant incubator (12 hours light and 23°C temperature) for 3 weeks, after which the roots were irrigated with 50 ml each of BTH (benzothiadiazole), serenade, N351 strain, valine, and a mixture of N351 strain and valine. Seven days after the treatment, the pathogen was inoculated, and the number of pathogens was investigated at 0 and 3 days, respectively. For pathogen counts, leaves were cut into 1 cm discs using a cork borer and then placed in e-tubes containing 1 ml of sterile water and beads. Leaves were homogenized at 6000 rpm for 2 min using a homogenizer (Bertin) and the pathogen counts in the juice were examined.

[0173] As a result, as shown in Figure 9a, Arabidopsis plants were treated with Pst pathogen and the number of pathogens was measured 3 days later. The number of pathogens in the N351 powder formulation and valine mixed treatment group was statistically significantly reduced the most compared to the untreated control group. The N351 powder formulation and valine mixed treatment group showed a superior pathogen suppression effect of 7.3 LogCFU / disc compared to 7.5 LogCFU / disc for N351 powder formulation and 8 LogCFU / disc for valine, and it was observed that the N351 strain, valine and their combination not only had antibacterial activity but also induced resistance.

[0174] Furthermore, to confirm the mechanism by which N351 powder formulation and valine mixed treatment induce induced resistance, the expression of disease resistance-related genes was investigated. Arabidopsis thaliana containing the PR (Pathogenesis-related protein) 1a promoter GUS fusion construct, a disease resistance marker gene, was used, and the preparation of Arabidopsis thaliana, N351 powder formulation and valine treatment, and pathogen treatment were performed in the same manner as above. Plant leaf samples to observe whether PR1a was expressed were prepared 0, 1, 2, and 3 days after plant disease treatment.

[0175] As a result, as shown in FIG. 9b, GUS activity was observed in the leaves from 1 to 3 days after the disease treatment in the N351 powder formulation and valine mixed treatment group. GUS expression was also observed from 1 to 3 days after the disease treatment in the N351 powder formulation alone treatment group, but the expression was stronger in the mixed treatment group. In the valine alone treatment group, GUS expression was most strongly induced 1 day after the disease treatment, and expression weakened as time passed. When the N351 powder formulation and valine mixed treatment were performed, strong GUS expression was observed to occur from the early stage with valine, and the persistence was maintained with the N351 powder formulation. EXAMPLES

[0176] Examination of growth promoting ability of N351 powder formulation and valine mixed treatment group The growth of soybean and cucumber was observed after mixing N351 powder formulation with valine.

[0177] One week after sowing in the pots, each plant was drenched with 5 ml of BTH, valine, N351 powder formulation, and a mixture of N351 powder formulation and valine. BTH, valine, N351 powder formulation, and a mixture of N351 powder formulation and valine were drenched twice at one-week intervals. One week after the last treatment, the stems of each treatment group were cut and weighed.

[0178] As a result, in soybeans, as shown in Figure 10a, the growth promotion was 9.1g, 9.3g, and 10.1g on average for N351 powder formulation, valine, and the mixture of N351 powder formulation and valine, respectively, which was 0.9g, 1.1g, and 1.9g higher than the untreated group (8.2g). In contrast, the growth of the BTH-treated group, which is the control group for the induced resistance product, was rather suppressed compared to the untreated control group. It was confirmed that all groups except the BTH-treated group showed statistically significant growth promotion compared to the untreated control group.

[0179] In cucumbers, as shown in Figure 10b, the average growth promotion was 15.5g, 14g, and 15.6g for N351 powder formulation, valine, and N531 powder formulation and valine mixture, respectively, which was 2.9g, 1.4g, and 3g higher than the untreated group (12.6g). As in soybeans, the growth promotion effect was the best in the mixed treatment group, but the strain was thought to have a greater effect than valine. The BTH treatment group also showed growth inhibition compared to the untreated control group in cucumbers. The N351 powder formulation and valine mixed treatment group induced statistically significant growth promotion.

[0180] Benzo(1,2,3)-thiadiazile-7-carbothioic acid S-methyl ester, benzothiadiazole (BTH) is known to be the most effective derivative that induces a pathogen resistance response in plants. However, treatment with BTH has serious problems in that it inhibits plant growth and significantly reduces yields.

[0181] From the above results, it was confirmed that N351 and the mixed treatment group of N351 and valine did not suppress plant growth but rather promoted plant growth.

[0182] From the above description, a person skilled in the art to which the present application pertains will understand that the present application can be implemented in other specific forms without changing its technical ideas or essential features. It should be understood that the above examples are merely illustrative and not limiting. This application should be interpreted as including all modifications or alterations derived from the meaning and scope of the claims and their equivalent concepts, rather than the specification.

[0183] JPEG2025511087000007.jpg190145

Claims

1. A composition for enhancing resistance to plant diseases, comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

2. 2. The composition for enhancing plant disease resistance according to claim 1, wherein the plant disease is at least one selected from the group consisting of Phytophthora stem and root rot, bacterial leaf blight, bacterial soft rot, rice blast, gray mold disease, leaf rust, powdery mildew, anthracnose, bacterial speck, damping off, Sclerotinia rot, Fusarium wilt, and wild fire.

3. The composition for enhancing plant disease resistance according to claim 1 , wherein the composition is for at least one selected from the group consisting of beans, fruit vegetables, leafy vegetables, root vegetables, and grains.

4. The plant diseases include Phytophthora sojae, Pseudomonas syringae pv. lachrimans, Pectobacterium carotovorum subsp. Carotovorum, Magnaporthe oryzae, Botrytis cinerea, Puccinia recondite, Blumeria graminis f. sp. hordei, Colletotrichum coccodes, Pseudomonas syringae pv. The composition for enhancing plant disease resistance according to claim 1, which is induced by at least one fungus selected from the group consisting of Pseudomonas syringae pv. tomato, Rhizoctonia solani, Pythium ultimum, Colletotrichum truncatum, Colletotrichum acutatum, Sclerotinia sclerotiorum, Phytophthora capsici, Phytophthora infestans, Fusarium oxysporum, and Pseudomonas syringae pv. tabaci.

5. The composition for enhancing resistance to plant diseases according to claim 1 , which is used to prevent or treat a plant disease.

6. The composition for enhancing plant disease resistance according to claim 1 , further comprising valine.

7. The composition for enhancing plant disease resistance according to claim 1 , which induces induced systemic resistance in a plant.

8. The composition for enhancing plant disease resistance according to claim 1 , which induces promotion of plant growth.

9. A composition for controlling plant diseases comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof.

10. The composition for controlling plant diseases according to claim 9 , further comprising valine.

11. A method for enhancing plant disease resistance, comprising the step of treating a plant body with the composition for enhancing plant disease resistance according to claim 1.

12. The method for enhancing plant disease resistance according to claim 11, wherein the plant is at least one selected from the group consisting of beans, fruit vegetables, leafy vegetables, root vegetables, and cereals.

13. A method for preventing or treating a plant disease, comprising the step of treating a plant body with the composition for enhancing plant disease resistance according to claim 1.

14. A method for controlling plant diseases, comprising the step of treating a plant body with the composition for controlling plant diseases according to claim 9.

15. Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P.

16. The microorganism according to claim 15, which has at least one of an ability to enhance resistance to a plant disease and an ability to control a plant disease.

17. Use of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P for enhancing resistance to plant diseases.

18. Use of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P for controlling plant diseases.

19. Use of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P for preventing or treating a plant disease.

20. Use of a composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof and valine for enhancing plant disease resistance.

21. Use of a composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof and valine for controlling plant diseases.

22. Use of a composition comprising at least one of the Bacillus amyloliquefaciens microorganism deposited under accession number KCCM13138P and a culture thereof, and valine, for preventing or treating a plant disease.

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