Strains belonging to the genus Bacillus and microbial pesticides using said strains

Abstract

Abstract:　Problem: To provide a soil-transmissible plant disease control agent or similar that can safely be used as a biological control agent.　Solution: A soil-transmissible plant disease control agent or similar, such as a vegetable bacterial wilt control agent, can be provided by using, as an active ingredient, live bacterial cells of a novel strain of a bacterium of the Bacillus or Fictibacillus genus with a heretofore unknown soil-transmissible plant disease controlling action, or a cultured product including said live bacterial cells. This control agent is characterized by being able, through a foliage application treatment, to control bacterial wilt and / or bacterial soft rot caused by the Ralstonia solanacearum species complex, which are soil-borne bacteria.　Selected drawing: none

Description

[Technical Field]

[0001] The present invention relates to a soil-borne plant disease control agent that is effective against bacterial wilt and other diseases caused by root infection when sprayed above ground, and / or a bacterial strain of the genus Bacillus or the like that can be cultured in a shorter time than conventional strains. [Background technology]

[0002] Controlling agricultural crop diseases (plant diseases) caused by pathogenic bacteria is a very important issue for agricultural workers around the world, and measures to address this issue include cultural control and the use of disinfectants, etc. However, there are some bacterial plant diseases that are difficult to control even with such cultural control and disinfectants.

[0003] For example, vegetable bacterial wilt is a soil-borne disease caused by the Ralstonia solanacearum species complex, a bacterium that lives in the soil. When tomatoes, eggplants, peppers, potatoes, etc. are infected with this bacterial wilt bacterium, the entire plant rapidly wilts and eventually dies, significantly affecting crop productivity. Furthermore, this bacterium is distributed throughout the world, mainly in tropical, subtropical, and temperate regions, and more than 200 species of crops, mainly the solanaceae plants mentioned above, are infected and damaged, making bacterial wilt a serious and important problem in agriculture.

[0004] Many methods for controlling bacterial wilt have been devised to date, including the use of chemical pesticides, soil disinfection using sunlight or reduction disinfection, and cultivating resistant rootstocks. However, even if bacteria on the soil surface are killed by soil disinfection, the bacteria can survive for long periods of time deep in the soil (approximately 50 cm to 1 m). Since there are no soil fungicides that can reach such depths, it is difficult to completely eliminate the bacteria from the soil. Furthermore, resistance is not complete for resistant varieties, and depending on environmental conditions, the effectiveness may be insufficient. Furthermore, although bacterial wilt resistance inducers (Patent Document 1) have been proposed, these methods are not sufficiently effective.

[0005] For this reason, in recent years, methods using biological pesticides have been proposed as a control method for suppressing disease in plants through antagonistic effects under conditions where pathogens are present in the soil. Regarding bacterial wilt disease, microbial pesticides using strains of Pseudomonas and Bacillus have been investigated for control (Non-Patent Document 1), but these biological pesticides have not been sufficiently effective. Attempts have also been made to control bacterial wilt disease by adding compost containing multiple microorganisms, but their effectiveness is unclear and control has often failed. While these methods have been shown to be effective by applying them to the soil, these methods require the establishment of antagonistic bacteria in the soil, rhizosphere, and plant body. However, consistently achieving stable efficacy in soil under highly variable conditions is often difficult. Furthermore, the establishment and proliferation of non-pathogenic bacterial strains in plant body is often difficult, which is one of the reasons why control methods using biological pesticides have not been established.

[0006] In this technical background, there has been a strong demand in the art for the development and commercialization of soil-borne disease control agents and the like which not only have sufficient basic activity but also exert sufficient (practical and stable) control effects against soil-borne plant diseases that have been difficult to control in the past, such as bacterial wilt, and which can be used safely as agricultural chemicals, using microorganisms that can establish and grow in soil or plant bodies without exhibiting pathogenicity within plant bodies. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Japanese Patent Application Laid-Open No. 2012-211124 [Non-patent literature]

[0008] [Non-Patent Document 1] Chemistry and Biology Vol.51(2013)No.8 pp.541~547 Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention has been made for the purpose of providing a non-phytopathogenic strain that can be safely used as a biological pesticide and that exhibits stable soil-borne plant disease control effects in actual crop production sites, as well as a soil-borne plant disease control agent and the like that uses the strain. In addition, because bacterial wilt is a disease caused by bacteria living in the soil, soil treatments such as irrigation and soil fumigation are usually used to treat it, but as mentioned above, soil treatments have several disadvantages. The present invention has been made with the objective of providing a new and effective processing method and constituent elements to overcome this problem. [Means for solving the problem]

[0010] In order to achieve the above-mentioned object, the present inventors have conducted extensive research and have focused on microbial treatment among various treatments. As a result of further intensive research, they have discovered for the first time that a novel strain of the genus Bacillus, which was previously unknown, can be used to control bacterial wilt, a soil-borne disease, sufficiently effectively by treating the above-ground parts of crops, for example, by treating the stems and leaves, rather than by soil treatment, or by combining this with soil treatment, while applying the strain indirectly, so to speak, to the application site. In other words, to achieve the above-mentioned object, the present inventors conducted extensive research and discovered that a strain belonging to the genus Bacillus, which was previously unknown, can be sprayed onto plants to exert practical effects in controlling soil-borne plant diseases without direct contact, and that the strain can be safely used in actual crop production sites. Furthermore, the present inventors discovered that the strain can be cultured in a shorter time than conventional plant disease control agents containing Bacillus bacteria, and that a control agent containing the strain as an active ingredient can be rapidly produced in a shorter time, leading to the completion of the present invention.

[0011] That is, the embodiments of the present invention are as follows. (1) A new bacterial strain, Bacillus sp. G4L1 (NITE BP-03204). (2) A novel bacterial strain, Bacillus timonensis G5S1 strain (NITE BP-03206). (3) A novel bacterial strain, Fictibacillus solisalsi G5L2 strain (NITE BP-03205). (4) The strain according to any one of (1) to (3), characterized in that it can control plant diseases by applying it to at least one of plant seeds, roots of plants, stems and leaves of plants, cultivation carriers, nutrient solutions, and soil. (5) The strain according to any one of (1) to (3), characterized by a fast growth rate. (6) A plant disease control agent characterized by containing, as an active ingredient, one or more strains according to any one of (1) to (3) or a culture of said strains. (7) The agent according to (6), characterized in that it can control plant diseases by applying it to at least one of the seeds of a plant, the roots of a plant, the stems and leaves of a plant, a cultivation carrier, a nutrient solution, and soil. (8) The agent according to (6) or (7), characterized in that it can control at least one disease of vegetables, fruit trees, rice, and wheat. (9) The agent according to any one of (6) to (8), which is capable of controlling diseases of Solanaceae plants and / or Brassicaceae plants. (10) The agent according to any one of (7) to (9), which can control soil-borne plant diseases by applying it to the stems and leaves of a plant body. (11) The agent according to (10), which is capable of controlling bacterial wilt and / or soft rot of vegetables. (12) A method for controlling plant diseases, comprising the step of contacting or mixing live cells of one or more strains according to any one of (1) to (3) or a culture containing the live cells with or into plant seeds, roots or above-ground parts of the plant, a cultivation carrier, a nutrient solution, or soil. (13) The method according to (12), characterized in that at least one disease of vegetables, fruit trees, rice, and wheat is controlled. (14) The method according to (12) or (13), characterized in that it controls diseases of Solanaceae plants and / or Brassicaceae plants. (15) The method according to any one of (12) to (14), characterized in that a soil-borne plant disease is controlled by applying the composition to the stems and leaves of a plant body. (16) The method according to any one of (12) to (15), characterized in that it controls vegetable bacterial wilt and / or vegetable soft rot.

[0012] Another embodiment of the present invention is as follows. (17) A novel bacterial strain, Bacillus sp. G4L1 strain (NITE BP-03204). (18) A novel bacterial strain, Bacillus timonensis G5S1 strain (NITE BP-03206). (19) A novel bacterial strain, Fictibacillus solisalsi G5L2 strain (NITE BP-03205). (20) The strain according to any one of (17) to (19), which can control soil-borne plant diseases by applying it to the stems and leaves of a plant, and which has a fast growth rate. (21) A plant disease control agent comprising one or more strains according to (17) to (20) or a culture of said strains as an active ingredient. (22) The agent according to (21), which is a soil-borne plant disease control agent. (23) The agent according to (22), which is an agent for controlling bacterial wilt of vegetables. (24) The agent according to any one of (21) to (23), which is an agent for controlling diseases of solanaceous plants. (25) A method for controlling plant diseases, comprising the step of contacting a plant body and / or soil (particularly rhizosphere soil) with live cells of one or more strains according to (17) to (20) or a culture containing the live cells. (26) The method according to (25), characterized in that it controls soil-borne plant diseases. (27) The method according to (26), characterized in that it controls bacterial wilt of vegetables. (28) The method according to any one of (25) to (27), characterized in that it is used to control diseases of solanaceous plants. (29) A method for controlling vegetable bacterial wilt, which is a soil-borne plant disease, characterized by achieving a control effect simply by spraying the vegetable bacterial wilt control agent according to (23) on the above-ground parts of a plant body. (30) The strain according to any one of (17) to (20), characterized in that it has a fast growth rate, and the culture time required to obtain an absorbance measurement value at 600 nm of 3.00 to 3.50 is (approximately) 10 hours for the strain, while it takes (approximately) 15 hours or more for conventional Bacillus bacteria, and thus grows in a short time. (31) The strain according to (30), characterized in that the culture time required to obtain an absorbance measurement value at 600 nm of 3.00 to 3.50 is 9 to 12 hours, specifically 9 to 11 hours, and even more specifically 9 to 10 hours, whereas conventional Bacillus bacteria require 14 to 16 hours. (32) A method for rapidly producing the control agent according to (21) or (22), characterized by culturing a fast-growing strain according to any one of (17) to (20), (30), and (31), obtaining the strain and / or a culture thereof in a short period of time, and producing a control agent containing the strain or the culture thereof as an active ingredient in a short period of time. [Effects of the Invention]

[0013] According to the present invention, it is possible to provide a soil-borne plant disease control agent that, when sprayed onto aboveground parts, shows a remarkable effect (practical and stable effect) in controlling soil-borne plant diseases that have been difficult to control in the past, such as bacterial wilt, and that can be safely used in actual crop production sites and can be cultured in a shorter time than conventional soil-borne plant disease control agents containing the genus Bacillus. [Brief explanation of the drawings]

[0014] [Figure 1] 1 shows a photograph of the plot treated with the G4L1 strain 14 days after inoculation with the tomato bacterial wilt pathogen in the test to confirm the effect of controlling bacterial wilt of tomato in Example 1. The test was carried out in five replicate pots to compare individual differences. [Figure 2] 1 shows a photograph of an untreated plot taken 14 days after inoculation with the tomato bacterial wilt pathogen in the test for confirming the effect of controlling bacterial wilt of tomato in Example 1. [Figure 3] The inoculation method of Example 2 is shown below. [Figure 4] The growth curves of the G4L1 strain and four other Bacillus strains are shown. DETAILED DESCRIPTION OF THE INVENTION

[0015] To achieve the above-mentioned object, the present inventors conducted extensive research and eventually focused on microbial treatment, searching extensively for microorganisms. As a result, they discovered for the first time that the bacterial strain they successfully isolated was a previously unknown novel strain, and that it had a novel and useful effect of antagonizing the causative bacterium of bacterial wilt disease. Furthermore, as a result of further research into application methods, they unexpectedly discovered that bacterial wilt disease can be controlled indirectly by spraying the soil bacterium, R. wilt, on the above-ground parts of plants, such as the stems and leaves, rather than by soil treatment.

[0016] That is, the present invention was finally completed as a result of further research into the isolation of a new bacterial strain, the use of said strain to control bacterial wilt disease, and the discovery of new conditions for control by spraying said strain on stems and leaves, and further organically combining these.

[0017] First, in the present invention, a strain belonging to the genus Bacillus or Fictibacillus is used as an active ingredient in a soil-borne plant disease control agent or the like. In the present invention, "soil-borne plant disease" refers to damage caused by pathogens infecting the underground parts of useful plants. This damage is mainly caused by pathogens that live in the soil, but is not limited to this. In this specification, although the control of bacterial wilt will be mainly described below, the present invention also has an effective effect on soil-borne plant diseases such as soft rot disease.

[0018] In particular, all of them are strains isolated from tomato leaves at 1-1 Yanido, Gifu City, Gifu Prefecture (within the campus of Gifu University). When identified by bacteriological properties and phylogenetic analysis based on the genomic sequence of the 16S rRNA gene, it was revealed that they are new strains belonging to Bacillus and Fictibacillus, which are non-pathogenic to plants. It is more preferable to use the Bacillus sp. G4L1 strain, Bacillus timonensis G5S1 strain, Fictibacillus solisalsi G5L2 strain, or a mutant of any of these strains that retains the same properties as the strain. From the tests conducted using the API20NE kit (manufactured by Sysmex Biomerieux Co., Ltd.), it has been revealed that the G4L1 strain, G5S1 strain, and G5L2 strain have the following bacteriological properties, respectively.

[0019] <G4L1 strain> The mycological properties of the G4L1 strain are as follows.

[0020] (A) Morphological properties Morphology: Bacillus Size: 0.9 - 1.0 μm × 2.0 - 8.0 μm Motility: None (B) Cultural properties Colony color: Light yellow to light greenish yellow Colony morphology: Irregular, elevation: Flat, periphery: Wavy (C) Physiological properties Gram stainability: Positive Optimum growth pH: Neutral range (pH 6.5 - 8.0) Optimum growth temperature: 35 °C Reduction of nitrate: + Indole production (tryptophan): - Glucose fermentation: - Arginine dihydrolase:+ Urease:- Hydrolysis (β-glucosidase):+ Hydrolysis (protease):+ Anabolic (glucose):+ Assimilation (arabinose):+ Assimilation (Mannose):+ Anabolic (mannitol):+ Anabolic (N-acetyl-glucosamine):+ Assimilation (Maltose):+ Assimilation (potassium gluconate):+ Anabolic (Capric Acid):- Assimilation (adipic acid): + Assimilation (Malate):+ Assimilation (trisodium citrate):+ Anabolic (phenyl acetate):+ This strain not only has an excellent bacterial wilt control activity, but also has the unique ability to control soil-borne plant diseases such as bacterial wilt and soft rot simply by treating the above-ground parts of plants.

[0021] Furthermore, the G4L1 strain was subjected to genome analysis for species identification. The genome analysis method was as follows.

[0022] DNA extracted from the G4L1 strain was barcoded with 400-base pairs. A library was created using the IonPGM™ Hi-Q™ ViewOT2 kit-400 (Thermo Fisher Scientific), and templates were prepared using the Ion OneTouch™ 2 System (Thermo Fisher Scientific), a preprocessing device for next-generation sequencers. The resulting templates were sequenced using the Ion PGM™ next-generation sequencer with the Ion PGM Sequencing Hi-Q View kit (Thermo Fisher Scientific) and the Ion 318 Chip version 2 (Thermo Fisher Scientific). After confirming the quality of the resulting sequences, adapters were removed. De novo assembly was performed using the analysis software CLC Genomics Workbench version 12 (QIAGEN), and the genome sequence was decoded. The obtained sequences were identified by ANI values ​​based on whole-genome comparison with closely related species using the TrueBac ID system version 1.92 (Chunlab).

[0023] In addition, digital DNA-DNA hybridization was performed with closely related reference strains and type strains using the BLAST+ method (Camacho et al. 2009) on the Genome-to-Genome Distance Calculator ver. 2.1 web server (Leibniz Institute DSMZ), and GGDC (formula 2) values ​​were calculated.

[0024] The genomic analysis results are shown in Table 1. The ANI values were all 95% or less compared with any of the related reference strains and the type strain, and the GGDC (formula 2 values) with the related reference strains and the type strain were also 27.3 - 56.3%. According to Reference 1 (Proc. Natl. Acad. Sci. USA November 10, 2009 106(45) 19126 - 19131), those with ANI values of 95% or less are regarded as new species, and according to Reference 2 (Stand Genomic Sci February 28, 2010, 2(1) 117 - 134), Reference 3 (Int J Syst Evol Microbiol January 1, 2007, 57(1) 81 - 91), those with GGDC (formula 2 values) of 70% or less are regarded as new species. Therefore, the G4L1 strain is considered to be a new species.

[0025]

Table 1

[0026] <G5S1 strain> The mycological properties of the G5S1 strain are as follows.

[0027] (A) Morphological properties Morphology: Bacillus Size: 0.8 - 1.3 μm × 2.5 - 5.5 μm Motility: Present (B) Cultural properties Colony color: Light reddish yellow - light yellow (translucent) Colony morphology: Circular, elevation: Convex circular, margin: entire (C) Physiological properties Gram stainability: Positive Optimum growth pH: Neutral range (pH 6.5 - 7.5) Optimum growth temperature: 40°C Reduction of nitrate: + Indole production (tryptophan): - Glucose fermentation: - Arginine dihydrolase: - Urease: - Hydrolysis (β-glucosidase): + Hydrolysis (protease): - Assimilation (glucose): + Assimilation (arabinose): - Assimilation (mannose): - Assimilation (mannitol): - Assimilation (N-acetyl-glucosamine): + Assimilation (maltose): + Assimilation (potassium gluconate): - Assimilation (capric acid): - Assimilation (adipic acid): - Assimilation (malate): - Assimilation (trisodium citrate): - Assimilation (phenyl acetate): - This strain not only has an excellent effect on controlling bacterial wilt, but also has a unique effect of being able to control bacterial wilt and soft rot, which are soil-borne plant diseases, by only treating the above-ground part of the plant.

[0028] <G5L2 strain> The mycological properties of the G5L2 strain are as follows.

[0029] (A) Morphological properties Morphology: Bacillus Size: 0.8 - 1.3 μm × 3.4 - 9.0 μm Motility: Present (B) Cultural properties Colony color: Pale greenish yellow Colony morphology: Circular, raised: Convex in the center, margin: entire (C) Physiological properties Gram stainability: Positive Optimum growth pH: Neutral range (around pH 6.5) Optimum growth temperature: 40 °C Reduction of nitrate: - Indole production (tryptophan): - Glucose fermentation: - Arginine dihydrolase: - Urease: - Hydrolysis (β-glucosidase):+ Hydrolysis (protease):+ Anabolic (glucose):+ Assimilation (arabinose):+ Assimilation (Mannose):+ Anabolic (mannitol):+ Anabolic (N-acetyl-glucosamine):+ Assimilation (Maltose):+ Assimilation (potassium gluconate):+ Anabolic (Capric Acid):- Assimilation (adipic acid): + Assimilation (Malate):+ Assimilation (trisodium citrate):- Anabolic (phenyl acetate):+ This strain not only has an excellent bacterial wilt control activity, but also has the unique ability to control soil-borne plant diseases such as bacterial wilt and soft rot simply by treating the above-ground parts of plants.

[0030] The G4L1, G5S1, and G5L2 strains were all internationally deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) on April 20, 2020 (Reiwa 2), and their deposit numbers are NITE BP-03204, NITE BP-03206, and NITE BP-03205, respectively.

[0031] Any medium can be used to culture strains belonging to the genus Bacillus or Fictibacillus, as long as the strain can grow in that medium. Examples include common media such as bouillon medium, as well as media containing glucose, peptone, and yeast extract. In addition to liquid media, solid media such as agar-containing slants and plates may also be used.

[0032] Any carbon source that can be assimilated by strains belonging to the genus Bacillus or Fictibacillus can be used as the carbon source for the medium. Specific examples include various synthetic or natural carbon sources that can be utilized by strains belonging to the genus Bacillus, such as glucose, arabinose, mannose, starch hydrolysates, and molasses. Similarly, various synthetic or natural sources that can be utilized by the strains can be used as the nitrogen source for the medium, including organic nitrogen-containing substances such as peptone, meat extract, yeast extract, and soybean meal. Furthermore, trace nutrient sources such as inorganic salts (e.g., salt and phosphate), metal salts (e.g., calcium, magnesium, and iron), vitamins, amino acids, and nucleic acid-related substances can be added as needed, in accordance with conventional methods for microbial culture. Furthermore, various additives, such as antifoaming agents, can also be added as needed.

[0033] Strains belonging to the genus Bacillus or Fictibacillus can be cultured under aerobic conditions, such as by shaking culture or aeration culture. The culture conditions are not limited to these, but preferably include a temperature of 20 to 45°C, preferably 30 to 40°C, a pH of 5 to 8, preferably 6 to 7, and a culture period of 0.5 to 2 days, preferably 1 to 1.5 days.

[0034] One of the important characteristics of this strain is its rapid growth rate. The above culture period is set as a general guideline to ensure that a sufficient bacterial cell concentration is obtained, and is set at 0.5 days (i.e., 12 hours) or more. However, (depending on the culture scale, etc.) a practically acceptable bacterial cell concentration can also be obtained with a shorter culture period, for example, (approximately) 7 to 10 hours, so this short culture period is not excluded. The fact that the time required for culturing the active ingredient microorganism is short means that the microbial control agent of the present invention can be produced quickly. Therefore, the present invention also contributes to a rapid production method of the microbial control agent. Furthermore, the risk of contamination during the culture can be significantly reduced.

[0035] The strains belonging to the genera Bacillus and / or Fictibacillus cultivated as described above can be used as active ingredients in soil-borne plant disease control agents and the like in the form of a culture containing viable cells without cell isolation. Alternatively, viable cells can be separated from the culture by conventional methods, such as membrane separation or centrifugation, and washed as necessary. The cultured and isolated cells themselves or a processed product thereof (e.g., a mixture of cultured and isolated cells with other components) can be used as an active ingredient. Furthermore, the culture or isolated viable cells can be used in the form of a dried product dried by methods such as freeze-drying or spray-drying, or in the form of a liquid or solid dilution. They can also be used in the form of various formulations containing various additives, according to conventional methods for agricultural chemical formulations. Examples of such formulations include granules, emulsifiable concentrates, wettable powders, and flowables.

[0036] The concentration of live bacterial cells contained in the soil-borne plant disease control agent according to the present invention is not particularly limited as long as it exerts the desired effect. However, if the bacterial concentration is too low, sufficient results will not be obtained, and conversely, if the bacterial concentration is too high, the bacteria will be wasted. Therefore, when prepared and used as a liquid formulation, for example, a concentration of 1 × 10 5 ~1×10 11 The concentration can be adjusted appropriately within the range of cfu / ml, preferably 1 × 10 6 ~1×10 10 In the present invention, "cfu" means colony forming unit. Even when a culture is used, it may be appropriately designed according to the above-mentioned viable cell concentration. The bacterial concentration can be expressed in "cfu", but it can also be expressed by the absorbance at 600 nm (OD 600 ) can also be displayed as measured values. For example, OD 600 The measured value of 1.0 was 2.3 × 10 for Bacillus sp. G4L1 strain when cultured in NB medium at 30°C with shaking at 200 rpm for 24 hours. 7 cfu / ml, and Bacillus zimonensis G5S1 strain was 1.4 × 10 7 cfu / ml, and the F. solisarci G5L2 strain was 1.6 × 10 8This roughly corresponds to cfu / ml. However, this value may not always be reached depending on the state of the bacterial strain and culture conditions.

[0037] The present invention makes it possible to control plant diseases, such as bacterial wilt, which is caused by bacteria belonging to the plant pathogenic Ralstonia solanacearum species complex and is found in Solanaceae plants (e.g., eggplant, tomato, bell pepper, paprika, potato, etc.) and Cucurbitaceae plants (e.g., cucumber, bitter melon, etc.). In the present invention, "control" means avoiding plant diseases by preventing useful plants from becoming infected with the target soil-borne plant pathogen.

[0038] The plant disease control agent according to the present invention can be applied directly or diluted with water or the like before application. The method of application as a chemical is not particularly limited, and examples include direct application to crops or seeds by spraying or immersion, application to soil, addition to water or fertilizer added to crops or soil, and application to agricultural equipment. Of these, direct application to crops is more preferred. In this way, various soil-borne plant diseases can be suppressed by having the soil-borne plant disease control agent according to the present invention present on plant bodies such as roots, stems, leaves, and seeds, or in the soil where the plant is cultivated.

[0039] The present invention is unique in that it can control soil bacteria not by soil spraying but by foliar spraying, i.e., by treating the top of the crop. Soil treatment requires burdensome work such as irrigation and large amounts of chemicals, which poses the risk of causing pollution such as contaminating water and the surrounding environment, but foliar spraying significantly reduces such risks, making the present invention extremely superior. Since bacterial wilt is caused by soil bacteria, it is generally thought that it can be controlled by soil treatment. However, contrary to this common technical knowledge, the present invention has great technical value in that it can achieve sufficient bacterial wilt control effects by only applying a minimally invasive treatment to the above-ground parts of the plant, and economic success can be fully expected.

[0040] The application amount of the pesticide of the present invention varies depending on the target crop, target disease, application method, occurrence tendency, degree of damage, environmental conditions, and formulation used, and is therefore preferably adjusted appropriately and cannot be generally specified. However, an example is a method in which 30 ml to 1 L, preferably 50 ml to 1 L, of liquid formulation is applied per crop plant. The application time also varies depending on the target disease and formulation, but it is preferably applied appropriately between two weeks before and after planting. Furthermore, because the present invention is a microbial pesticide that does not involve concerns about the emergence of resistant bacteria, it can be used continuously for several days or for successive crops.

[0041] Furthermore, the soil-borne plant disease control agent and the like according to the present invention can be used, if necessary, in combination with other fertilizers and agricultural chemicals, such as fungicides, antiviral agents, insecticides, miticides, nematicides, synergists, attractants, herbicides, plant growth regulators, etc. In this case, they may be applied as a mixture under conditions that do not significantly affect the strain of bacteria that is the active ingredient, or they may be applied separately at intervals or both at the same time.

[0042] Next, examples of known fungicides (fungicidal active ingredients) or disease control compounds that may be mixed or used in combination are shown below, but are not limited to these examples.

[0043] Fungicidal active ingredient or disease control compound: Agrobacterium radiobacter, azaconazole, acibenzolar-S-methyl, azoxystrobin, anilazine, amisulbrom, aminopyrifen, ametoctradin, aldimorph, isotianil, isopyrazam, isofetamide, isofetamid, isoflucypram, isoprothiolane, ibuprofen, thiazolinone, thiazolinone, thiazolinone, thiazolinone Puconazole, ipflufenoquin, ipfentrifluconazole, iprodione, iprovalicarb, iprobenfos, imazalil, iminoctadine-albesilate, iminoctadine-triacetate, imibenconazole, inpyrfluxam, imprimatin A A), imprimatin B, edifenphos, etaconazole, ethaboxam, ethirimol, ethoxyquin, etridiazole, enestroburin, enoxastrobin, epoxiconazole, organic oiloils, oxadixyl, oxazinylazole, oxathiapiprolin, oxycarboxin, oxine-copper, oxytetracycline, oxpoconazole-fumarate, oxolinic acid, copper octanoate, octhilinone, ofurace, orysastrobin, o-phenylphenol, kasugamycin, captafol, carpropamid, carbendazim, carboxin, carvone, Candida oleophila oleophila, Candida saitoana, quinoxyfen, quinofumelin, chinomethionat, captan, quinconazole, quintozene, guazatine, cufraneb, coumethoxystrobin, coumoxystrobin, Gliocladium catenulatum, Cryptococcus albidus, kresoxim-methyl, clozylacon, Clonostachys rosea rosea), chlozolinate, chloroinconazide, chlorothalonil, chloroneb, Chaetomium cupreum, ConioscilliumConiothyrium minitans, cyazofamid, diethofencarb, diclocymet, dichlofluanid, dichlobentiazox, diclomezine, dicloran, dichlorophen, dithianon, diniconazole, diniconazole-M , zineb, dinocap, dipymetitrone, diphenylamine, difenoconazole, cyflufenamid, diflumetorim, cyproconazole, cyprodinil, simeconazole, dimethirimol, dimethyl disulfide disulfide, dimethomorph, cymoxanil, dimoxystrobin, Pseudozyma flocculosa, Pseudomonas aureofaciens, Pseudomonas chlororaphis, Pseudomonas syringae, Pseudomonas flurorescens, Pseudomonas rhodesiae, ziram, silthiofam, Zucchini yellow mosaic virus attenuated strain, streptomycin, Streptomyces griseoviridis griseoviridis), StreptomycesStreptomyces lygicus, spiroxamine, sedaxane, seboctylamine, zoxamide, solatenol, dazomet, Talaromyces flavus flavus, tiadinil, thiabendazole, thiram, thiophanate, thiophanate-methyl, thifluzamide, thiram, tecnazene, tecloftalam, tetraconazole, debacarb, tebuconazole, tebufloquin, terbinafine, dodine, dodemorph, triadimenol, triadimefon, triazoxide, trichlamide, triclopyricarb, Trichoderma asperellum asperellum, Trichoderma atroviride, Trichoderma gamsii, Trichoderma stromaticum, Trichoderma harzianum, Trichoderma viride, Trichoderma virens, Trichoderma polysporum, Trichoderma lignorumlignorum, tricyclazole, triticonazole, tridemorph, triflumizole, trifloxystrobin, triforine, tolylfluanid, tolclofos-methyl, tolnifanide, tolprocarb, nabam, natamycin, naftifine, nitrapyrin, nitrothal-isopropyl, nuarimol, copper nonyl phenol sulphonate, Paenibacillus polymyxa, Burkholderia Barkholderia cepacia, Bacillus amyloliquefaciens, Bacillus simplex, Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, harpin protein, Variovorax paradoxus, validamycin, valifenalate, Pantoea agglomerans, picarbutrazox, bixafen, picoxystrobin, Pythium oligandrum oligandrum, pydiflumetofen, bitertanol, binapacryl, hinokitiol, non-pathogenic Erwinia carotovoracarotovora, non-pathogenic Rhizobium vitis, biphenyl, piperalin, hymexazol, pyraoxystrobin, pyraclostrobin, pyraziflumid, pyrazophos, pyrapropoyne, pyrametostrobin, pyriophenone, pyrisoxazole, pyridaclomethyl, pyrifenox, pyributicarb, pyribencarb carb), pyrimethanil, pyroquilon, vinclozolin, ferbam, famoxadone, phenazine oxide oxide, fenamidone, phenaminestrobin, fenarimol, fenoxanil, ferimzone, fenpiclonil, fenpicoxamid, fenpyrazamine, fenbuconazole, fenfuram, fenpropidin, fenpropimorph, fenhexamid, folpet, phthalide, Fusarium oxysporum oxysporum, bupirimate, fuberidazole, blasticidin-S, furametpyr, furalaxyl, furancarboxylic acid, fluazinam, fluindapyr, fluoxastrobin, fluoxapiprolin, fluoxytioconazole, fluopicolide, fluopimomide, fluopyram, fluoroimide, fluxapyroxad, fluquinconazole, fluconazole, fluconazole-cis-cis), fludioxonil, flusilazole, flusulfamide, flutianil, flutolanil, flutriafol, flufenoxadiazam, flufenoxystrobin, flubeneteram, flumethylsulforim, flumetover, flumorph, Phlebiopsis giganteagigantea, proquinazid, prochloraz, procymidone, prothiocarb, prothioconazole, bronopol, propamocarb hydrochloride, propiconazole, propineb, probenazole, bromuconazole, flometoquin, florylpicoxamid, hexaconazole, benalaxyl, benalaxyl M (benalaxyl-M), benodanil, benomyl, pefurazoate, penconazole, pencycuron, benzovindiflupyr, benthiazole, benthiavalicarb-isopropyl, penthiopyrad, penflufen, boscalid, fosetyl (including salts of aluminum, calcium, sodium, etc.), polyoxin, polycarbamate, Bordeaux mixture mixture), mancopper, mancozeb, mandipropamid, mandestrobin, maneb, myclobutanil, Mitsuaria chitosanitabida, mineral oiloils, mildiomycin, methasulfocarb, metam, metalaxyl, metalaxyl-M, metarylpicoxamid, metiram, methyltetraprole, metconazole, metominostrobin, metrafenone, mepanipyrim, mefentrifluconazole, meptyldinocap, mepronil, iodocarb, laminarin, phosphorous acid and salts, basic copper chloride oxychloride, silver, copper(II) acetate, cuprous oxide, copper hydroxide, potassium bicarbonate, sodium bicarbonate, sulfur, oxyquinoline sulfate, copper sulfate, (3,4-dichloroisothiazol-5-yl)methyl 4-(tert-butyl)benzoate (chemical name, CAS registration number: 1231214-23-5), UK-2A (code number), dodecylbenzenesulfonate bis(ethylenediamine)copper complex salt [II] (DBEDC), triphenyltin acetate (TPTA), triphenyltin chloride (TPTC), triphenyltin hydroxide (TPTH).

[0044] Next, examples of known insecticides (insecticidal active ingredients), acaricides (acaricidal active ingredients), nematicides (nematicidal active ingredients), and synergist compounds (synergist active ingredients) that may be mixed or used in combination are shown below, but they are not limited to these examples.

[0045] Insecticidal, acaricidal, nematicidal and synergistic active ingredients: Acrinathrin, azadirachtin, azamethiphos, acinonapyr, azinphos-ethyl, azinphos-methyl, acequinocyl, acetamiprid, acetoprole, acephate, azo Azocyclotin, abamectin, afidopyropen, afoxolaner, amidoflumet, amitraz, alanycarb, aldicarb, aldoxycarb, allethrin (including d-cis-trans- and d-trans-isomers) [mu], isazophos, isamidofos, isocarbophos, isoxathion, isocycloseram, isofenphos-methyl, isoprocarb, epsilon-metofluthrin, epsilon-momfluorothrin omfluorothrin, ivermectin, imicyafos, imidacloprid, imiprothrin, indoxacarb, esfenvalerate, ethifencarb, ethion, ethiprole, ethylene dibromide, etoxazole, etofenprox, ethoprophos, etrimfos, emamectin, emamectin benzoatebenzoate, endosulfan, empenthrin, oxazosulfyl, oxamyl, oxydemeton-methyl, oxydeprofos, omethoate, nuclear polyhedrosis virus, cadusafos, kappa-tefluthrin, kappa-bifenthrin, karanjin, cartap, Granulosis virus virus), carbaryl, carbosulfan, carbofuran, gamma-BHC, xylylcarb, quinalphos, kinoprene, chinomethionate, Entero virus, coumaphos, cryolite, clothianidin, clofentezine, chromafenozide, chlorantraniliprole, chlorethoxyfos, chlordane, chloropicrin, chlorpyrifos, chlorpyrifos-methyl, chlorfenapyr, chlorfenvinphos, chlorfluazuron, chlormephos, chloroprallethrin, Entomopoxi virus, Irido virusvirus), cyazypyr, cyanophos, diafenthiuron, diamidaphos, cyantraniliprole, cyetpyrafen, dienochlor, cyenopyrafen, dioxabenzofos, diofenolan, Sigma virusvirus), cyclaniliprole, cycloxaprid, dicrotophos, diclofenthion, cyclobutrifluram, cycloprothrin, dichlorvos, dichloromezotiaz, dicofol, dicyclanil, disulfoton, dinotefuran, dinobuton, cyhalodiamide, cyhalothrin [including gamma- and lambda-forms], cyphenothrin [(1R)-trans- α-, β ... acid), methyl jasmonate, silafluofen, cyromazine, Steinernema carpocapsae, Steinernema kushidai, Steinernema glaceraiglaseri, spidoxamat, spinetoram, spinosad, spirodiclofen, spirotetramat, spiropidione, spiromesifen, sulcofuron sodium, sulfluramid, sulfoxaflor, sulfote sulfotep, diazinon, thiacloprid, thiamethoxam, tioxazafen, thiodicarb, thiocyclam, thiosultap, thionazin, thiofanox, thiometon, tyclopyrazoflor, tetracycline, Chlorantraniliprole, tetrachlorvinphos, tetradifon, tetraniliprole, tetramethylfluthrin, tetramethrin, tebupirimfos, tebufenozide, tebufenpyrad, te Tefluthrin, teflubenzuron, demeton-S-methyl, temephos, deltamethrin, terbufos, tralomethrin, transfluthrin, triazamate, triazophos, trichlorfon, Trichoderma asperellum, TrichodermaTrichoderma harzianum, triflumuron, triflumezopyrim, trimethacarb, tolfenpyrad, naled, nicotine, nicofluprole, nitenpyram, nemadectin, Denso virus, novaluron, noviflumuron, Paecilomyces lilacinus, Burkholderia cepacia, Burkholderia rinojensis, Verticillium lecanii, hydroprene, Pasteuria Pasteuria nishizawae, Pasteuria penetrans, Bacillus thuringiensis, insect toxins produced by Bacillus thuringiensis, Bacillus thuringiensis subsp. Aizawai, Bacillus thuringiensis subsp. Israelensis, Bacillus thuringiensis subsp. Kurstaki, Bacillus thuringiensis subsp. Tenebrionis, Bacillus popilliae, Bacillus licheniformislicheniformis, vamidothion, parathion, parathion-methyl, halfenprox nprox, halofenozide, bioallethrin, bioallethrin S-cyclopentenyl, bioresmethrin, bis-(2-chloro-1-methylethyl) ether (DCIP), bistrifluron, hydramethylnon, bifenazate, bifenthrin, pyflubumide, piperonyl butoxide butoxide, pymetrozine, pyraclofos, pyrafluprole, pyridaphenthion, pyridaben, pyridalyl, pyrifluquinazon, pyriprole, pyriproxyfen, pirimicarb, pyrimidifen, pyriminostrobin, pirimiphos-methyl, pyrethrins thrine, famphur, fipronil, fenazaquin, fenamiphos, fenitrothion, fenoxycarb, fenothiocarb, phenothrin [including the (1R)-trans-isomer], fenobucarb, fenthion, phenthoate, fenvalerate, fenpyroximate, fenbutatin oxide, fenpropathrin, fonofos, sulfuryl fluoridefluoride, butocarboxim, butoxycarboxim, buprofezin, furathiocarb, prallethrin, fluacrypyrim, fluazaindolizine, fluazuron, fluensulfone, fluopyram, fluoroacetic acid sodium saltfluoroacetate, fluxametamide, flucycloxuron, flucythrinate, flusulfamide, fluthrin, fluvalinate [including tau-form], flupyradifurone, flupyrazofos, flupirimin, flufiprole, flufenerim, flufenoxystrobin, flufenoxuron, fluhexafon, flubendiamide, flupentiophenox enox, flumethrin, fluralanal, flurimfen, prothiofos, protrifenbute, flonicamid, propaphos, propargite, prohydrojasmon, profenofos, broflanilide, profluthrin, propetamphos, propoxur, flometoquin, bromopropylate, hexythiazox, hexaflumuron, and pekiromyces Paecilomyces tenuipes, Paecilomyces fumosoroceus, Paecilomyces lilacinuslilacinus, heptafluthrin, heptenophos, permethrin, benclothiaz, benzpyrimoxan, bensultap, benzoximate, bendiocarb, benfuracarb, Pochonia chlamydosporia, Beauveria tenella, Beauveria bassiana, Beauveria brongniartii brongniartii, phoxim, phosalone, fosthiazate, fostietan, phosphamidon, phosmet, polynactins, formetanate, phorate, machine oil, malathion, milbemectin, mecarbam, mesulfenphos, methomyl, metaldehyde, metaflumizone, methamidophos, metham, methiocarb, methidathion, methyl isothiocyanate, methyl bromide bromide, methoxychlor, methoxyfenozide, methotrin, metofluthrin, methoprene, metolcarb, mevinphos, meperfluthrin, Monacrosporium phymatopagumphymatophagum, Monacrosporium phymatophagum, monocrotophos, momfluorothrin, Trichoderma harzianum, litlure-A, litlure-B, aluminum phosphide, zinc phosphide, hydrogen phosphine, lufenuron, rescale, resmethrin, lepimectin, rotenone, cytoplasmic polyhedrosis virus, fenbutatin oxide, calcium nitrogen cyanide, organotins, nicotine sulfate, (Z)-11-tetradecenyl acetate, (Z)-11-hexadecenal, (Z)-11-hexadecenyl acetate, (Z)-9,12-tetradecadienyl acetate, (Z)-9-tetradecen-1-ol, (Z,E)-9,11-tetradecadienyl acetate, (Z,E)-9,12-tetradecadienyl acetate, 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT), 1,3-dichloropropene, 2,4-dichloro-5-{2-[4-(trifluoromethyl)phenyl]ethoxy}phenyl 2,2,2-trifluoroethyl Sulfoxide (chemical name, CAS registration number: 1472052-11-1), 2,4-dimethyl-5-[6-(trifluoromethylthio)hexyloxy]phenyl-2,2,2-trifluoroethylSulfoxide (chemical name, CAS registry number: 1472050-34-2), 2-{2-fluoro-4-methyl-5-[(2,2,2-trifluoroethyl)sulfinyl]phenoxy}-5-(trifluoromethyl)pyridine (chemical name, CAS registry number: 1448758-62-0), 3-chloro-2-{2-fluoro-4-methyl-5-[(2,2,2-trifluoroethyl)sulfinyl]phenoxy}-5-(trifluoromethyl)pyridine (chemical name, CAS registry number: 1448761-28-1), 4,6-dinitro-o-cresol (DNOC), 4-fluoro-2-methyl-5-(5,5-dimethylhexyloxy)phenyl 2,2,2-trifluoroethyl Sulfoxide (chemical name, CAS registration number: 1472047-71-4), Bt proteins (Cry1Ab, Cry1Ac, Cry1Fa, Cry2Ab, mCry3A, Cry3Ab, Cry3Bb, Cry34 / 35Ab1), methyleugenol, 4-(p-acetoxyphenyl)-2-butanone, (Z)-10-tetradecenyl acetate, (E,Z)-4,10-tetradecadinyl acetate, (Z)-8-dodecenyl acetate, (Z)-11-tetradecenyl acetate, (Z)-13- Icosen-10-one, 14-methyl-1-octadecene, AKD-1193 (code number), BCS-AA10147 (code number), CL900167 (code number), O,O-diethyl-O-[4-(dimethylsulfamoyl)phenyl]-phosphorothioate (DSP), O-ethyl-O-4-(nitrophenyl)phenylphosphonothioate (EPN), RU15525 (code number), XMC (XMC), Z-13-icosen-10-one, ZXI8901 (code number), F4260 (code number).

[0046] Next, known herbicidal compounds, herbicidal active ingredients and plant growth regulator compounds that may be mixed or used in combination will be exemplified below, but the present invention is not limited to these examples.

[0047] Herbicide compounds or herbicidal active ingredients: Ioxynil (including its lithium salt, sodium salt, and salt with octanoic acid, etc.), aclonifen, acrolein, azafenidin, acifluorfen (including its sodium salt, etc.), azimsulfuron, asulam, acetochlor, atrazine, anisiflupurin, anilofos, amicarbazone, amidosulfuron, amitrole, aminocyclopyrachlor, aminopyralid, amiprofos-methyl, ametryn, Araujia mosaic virus Virus), alachlor, Alternaria destruensdestruens, alloxydim (including salts with sodium etc.), ancymidol, isouron, isoxachlortole, isoxaflutole, isoxaben, isodecyl alcohol ethoxylate, isoproturon, ipfencarbazone arbazone, imazaquin, imazapic (including salts with amines, etc.), imazapyr (including salts with isopropylamine, etc.), imazamethabenz, imazamethabenz-methyl, imazamox, imazethapyr, imazosulfuron, indaziflam ), indanofan, eglinazine-ethyl, esprocarb, ethametsulfuron-methyl, ethalfluralin, ethidimuron, ethoxysulfuron, ethoxyfen, ethoxyfen-ethyl, etof Ethofumesate, etobenzanid, epirifenacil, endothal disodium, oxadiazon, oxadiargyl, oxaziclomefone, oxasulfuron, oxyfluorfen, oryzalin, Obuda Pepper VirusVirus), orthosulfamuron, orbencarb, oleic acid, cafenstrole, caprylic acid, capric acid, carfentrazone-ethyl, karbutilate, carbetamide, quizalofop, quizalofop-ethyl, quizalofop-P-ethyl, quizalofop-P-tefuryl, Xanthomonas campestris, quinoclamine, quinclorac, quinmerac, citric acidacid, cumyluron, clacyfos, glyphosate (including salts such as sodium, potassium, amine, propylamine, isopropylamine, ammonium, isopropylammonium, guanidine, monoethanolamine, choline, BAPMA (N,N-bis-(aminopropyl)methylamine), dimethylamine, or trimesium), glufosinate (including salts such as amine or sodium), glufosinate-P, glufosinate-P-sodium, clethodim, clodinafop, clodinafop-propargyl, clopyralid (including monoethanolamine salt), clomazone, clomethoxyfen oxyfen, clomeprop, cloransulam-methyl, chloramben, chloridazon, chlorimuron, chlorimuron-ethyl, chlorsulfuron, chlorthal-dimethyl, chlorthiamid, chlorphthalim, chlorflurenol-methyl, chlorpropham, chlorbromuron, chloroxuron, chlorotoluron, ketospiradox (including salts such as sodium, calcium, or ammonia), colletotricham Colletotrichum orbiculare, Colletotrichum gloeosporioides, Colletotrichum truncateumtruncatum, Chondrostercum purpureum, saflufenacil, sarmentine, cyanazine, cyanamide, diuron, diethathyl-ethyl, dioxopyritrione, dicamba (amines, diethylamine, isopropylamine, diglycolamine, dimethylammonium, diolamine, isopropylammonium, olamine, potassium, trolamine, BAPMA (N,N-bis-(aminopropyl)methylamine), choline, salts such as sodium or lithium, or esters such as methyl esters), cycloate, cycloxydim , diclosulam, cyclosulfamuron, cyclopyranil, cyclopyrimorate, dichlobenil, diclofop, diclofop-P-methyl, diclofop-methyl, dichlorprop, dichlorprop-P (including dimethylammonium, potassium, sodium, choline salts, etc., or esters such as butotyl ester, 2-ethylhexyl ester, isoctyl ester, methyl ester), diquat, diquat dibromidedibromide, dithiopyr, siduron, dinitramine, cinidon-ethyl, cinosulfuron, dinozeb (including acetate), dinoterb, cyhalofop, cyhalofop-butyl, cypyrafluone, diphenamid, difenzoquat zoquat, diflufenican, diflufenzopyr, simazine, dimesulfazet, dimethachlor, dimethametryn, dimethenamid, dimethenamid-P, simetryn, dimepiperate, dimefuron, Pseudomonas Pseudomonas fluorescens, cinmethylin, swep, sulcotrione, sulfentrazone, sulfosate, sulfosulfuron, sulfometuron-methyl, sethoxydim, Scelerothinia minor, terbacil, daimuron, thaxtomin A, Tobacco Mild Green Mosaic Tobamovirus, Tobacco Rattle VirusVirus), dalapon, thiazopyr, thiafenacil, thiencarbazone (including sodium salt, methyl ester, etc.), thiocarbazil, thiobencarb, thidiazimin, thidiazuron, thifensulfuron, thifensulfuron-methyl, desmedipham, desmetryne, tetflupyrolim et), thenylchlor, tebutam, tebuthiuron, tepraloxydim, tefuryltrione, terbuthylazine, terbutryn, terbumeton, tembotrione, topramezone, tralkoxydim, triaziflam, triasulfuron, triafamone, triallate, tri Trietazine, triclopyr, triclopyr-butotyl, triclopyr-triethylammonium, tritosulfuron, tripyrasulfone, trifludimoxazin, triflusulfuron-methyl, trifluralin, trifloxysulfuron (including salts with sodium etc.), tribenuron-methyl, tolpyralate, naptalam (including salts with sodium etc.), naproanilide, napropamide, napropamide-M, nicosulfuron, lactic acid acid), neburon, norflurazon, Burkholderia rinojensis, vernolate, paraquat, paraquat dichloride dichloride, halauxifen, halauxifen-benzyl, halauxifen-methyl, haloxyfop, haloxyfop-P, haloxyfop-etotyl, haloxyfop-P-methyl, halosafen, halosulfuron-methyl, bixlozone, picloram (including salts with dichloroammonium, trollamine, etc.), picolinafen, bicyclopyrone,Bispyribac-sodium, pinoxaden, bipyrazone, bifenox, piperophos, pyraclonil, pyrasulfotole, pyrazoxyfen, pyrazosulfuron-ethyl, pyrazolynate, bilanafos, pyraflufen, pyrazolynate Flufen-ethyl, pyridafol, pyrithiobac-sodium, pyridate, pyriftalid, pyributicarb, pyribenzoxim, pyrimisulfan, pyriminobac-methyl, pyroxasulfone, pyroxsulam, Phytopsora Phytophthora palmivora, phenisopham, fenuron, fenoxasulfone, fenoxaprop (including methyl, ethyl, and isopropyl esters), fenoxaprop-P (including methyl, ethyl, and isopropyl esters), fenquinotrione, fenthiaprop-ethyl, fentrazamide, fenpyrazone, phenmedipham, Phoma chenopodicola, Phoma herbarum, Phoma macrostoma macrostoma), butachlor, butafenacil, butamifos,Butyrate, Puccinia canaliculata, Puccinia thraspeos thlaspeos, butenachlor, butralin, butroxydim, flazasulfuron, flamprop (including methyl, ethyl, and isopropyl esters), flamprop-M (including methyl, ethyl, and isopropyl esters), primisulfuron, primisulfuron-methyl, fluazifop-butyl, fluazifop-P, fluazifop-P-butyl, fluazolate, fluometuron, fluoroglycofen-ethyl, flucarbazone-sodium, fluchloralin lin), flucetosulfuron, fluthiacet-methyl, flupyrsulfuron-methyl (including salts such as sodium, calcium or ammonia), flufenacet, flufenpyr-ethyl, flupropanate (including sodium salt), flupoxame, flumioxazin, flumiclorac-pentyl, flumetsulam, fluridone, flurtamone, fluroxypyr (including esters such as butomethyl, meptyl, or salts such as sodium, calcium or ammonia), flurochloridone, pretilachlor,Procarbazone (including salts with sodium etc.), prodiamine, prosulfuron, prosulfocarb, propaquizafop, propachlor, propazine, propanil, propyzamide, propisochlor, propyrisulfuron, propham, profluazol, prohexadione-calcium, propoxycarbazone, propoxycarbazone-sodium salt e-sodium), profoxydim, bromacil, brompyrazone, prometryn, prometon, bromoxynil (including esters of butyric acid, octanoic acid, or heptanoic acid, etc.), bromofenoxim, bromobutide, florasulam, florpyrauxifen, florpyrauxifen-benzyl, hexazinone, pethoxamid, benazolin, benazolin-ethyl, penoxsulam, pepino Pepino Mosaic Virus, heptamaloxyloglucan, beflubutamid, beflubutamid-M, pebulate, pelargonic acid, bencarbazone, benquitrione, benzfendizone,bensulide, bensulfuron, bensulfuron-methyl, benzobicyclon, benzofenap, bentazone, pentanochlor, pendimethalin, pentoxazone, benfluralin, benfuresate, fosamine, fomesafen, foramsulfuron, forchlorfenuron, mecoprop (including salts of sodium, potassium, isopropylamine, triethanolamine, dimethylamine, diolamine, trollamine, choline, etc., or esters such as etadyl ester, 2-ethylhexyl ester, isoctyl ester, methyl ester, etc.), mecoprop Prop-P-potassium salt (mecoprop-P-potassium), mesosulfuron (including esters such as methyl), mesotrione, metazachlor, metazosulfuron, methabenzthiazuron, metamitron, metamifop, metam (including salts such as sodium), methane Disodium arsonic acid (DSMA), methiozolin, methyldymuron, metoxuron, metosulam, metsulfuron-methyl, metobromuron, metobenzuron, metolachlor, metribuzin, mepiquat chloride, mefenacet, monosulfuron (including methyl, ethyl, and isopropyl esters),Monolinuron, molinate, iodosulfuron, iodosulfuron-methyl-sodium, iofensulfuron, iofensulfuron-sodium, lactofen, en), lancotrione, linuron, rimisoxafen, rimsulfuron, lenacil, 2,2,2-trichloroacetic acid (TCA) (including salts such as sodium, calcium, or ammonia), 2,3,6-trichlorobenzoic acid (2,3,6-TBA), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 2,4-dichlorophenoxyacetic acid (2,4-D) (amines, diethylamine, triethanolamine, isopropyl alcohol, methyl methyl amine ... propylamine, dimethylammonium, diolamine, dodecylammonium, heptylammonium, tetradecylammonium, triethylammonium, tris(2-hydroxypropyl)ammonium, trolamine, choline, sodium or lithium salts, or esters such as butotyl ester, 2-butoxypropyl ester, 2-ethylhexyl ester, methyl ester, ethyl ester, butyl ester, isobutyl ester, octyl ester, pentyl ester, propyl ester, isoctyl ester, isopropyl ester, meptyl ester, tefuryl ester, etc.), 2,4-dichlorophenoxybutyric acid (2,4-DB) (amine, diethylamine, triethanolamine, isopropylamine, dimethylammonium, choline, sodium or lithium salts, or esters such as isoctyl ester), 2-amino-3-chloro-1,4-naphthoquinone (ACN), 2-methyl-4-chlorophenoxyacetic acid (MCPA) (sodium, dimethylammonium, choline salts, or 2-ethylhexyl ester, isoctyl ester, ethyl esters, etc.), 2-methyl-4-chlorophenoxybutyric acid (MCPB) (including sodium salt, ethyl ester, etc.), 4-(2,4-dichlorophenoxy)butyric acid (2,4-DB), 4,6-dinitro-O-cresol (DNOC) (including amine or sodium salts, etc.), (5S)-3-(3,5-difluorophenyl)-N-[rel-(3R,5R)-5-(trifluoromethylsulfonylcarbamoyl)tetrahydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamide ((5S)-3-(3,5-difluorophenyl)-N-[rel-(3R,5R)-5-(trifluoromethylsulfonylcarbamoyl)tetrahydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamid) (chemical name, CAS registration number: 2266183-40-6) (WO 2018 / 228986, WO 2020 / 114934), N4-(2,6-difluorophenyl)-6-(1-fluoro-1-methyl-ethyl)-1,3,5-triazine-2, 4-Diamine (N4-(2,6-difluorophenyl)-6-(1-fluoro-1-methyl-ethyl)-1,3,5-triazine-2,4-diamine) (chemical name, CAS registration number: 1606999-43-2) (WO 2014 / 064094, WO 2015 / 162164), (5S)-3-(3,5-difluorophenyl)-N-[(3R)-5-(methylsulfonylcarbamoyl)-2,3-dihydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamine (5S)-3-(3,5-difluorophenyl)-N-[(3R)-5-(methylsulfonylcarbamoyl)-2,3-dihydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamid) (chemical name, CAS registration number: 2266190-06-9) (WO 2018 / 228986, WO 2020 / 114934), (5R)-3-(3,5-difluorophenyl)-5-methyl-N-[rel-(3R,5R)-5-(methylsulfonylcarbamoyl)-2,3-dihydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamid (5R)-3-(3,5-difluorophenyl)-5-methyl-N-[rel-(3R,5R)-5-(methylsulfonylcarbamoyl)tetrahydrofuran-3-yl]-4H-isoxazole-5-carboxamid (chemical name, CAS registration number: 2266164-36-5) (WO 2018 / 228986, WO 2020 / 114934), (5R)-3-(3,(5R)-3-(3,5-difluorophenyl)-N-[(3R)-5-(methoxycarbamoyl)-2,3-dihydrofuran-3-yl]-5-methyl-4H-isoxazole-5-carboxamid (chemical name, CAS registration number: 2266170-31-2) (WO 2018 / 228986, WO 2018 / 228986, WO 2018 / 228986) 2020 / 114934), 2-[2-(3,4-dimethoxyphenyl)-6-methyl-3-oxo-pyridazine-4-carbonyl]cyclohexane-1,3-dione (chemical name, CAS registration number: 2138855-12-4) (WO 2017 / 178582, WO 2018 / 015476), 4-hydroxy-1-methyl-3- [4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one (4-hydroxy-1-methyl-3-[4-(trifluoromethyl)-2-pyridyl]imidazolidin-2-one) (chemical name, CAS registration number: 1708087-22-2) (WO 2015 / 059262, WO 2018 / 015476), 6-(1-fluorocyclopentyl)-N4-(2,3,5,6-tetrafluorophenyl)-1,3,5-triazine-2,4-diamine (6-(1-fluoro cyclopentyl)-N4-(2,3,5,6-tetrafluorophenyl)-1,3,5-triazine-2,4-diamine) (chemical name, CAS registration number: 1820807-75-7) (WO 2015 / 162164), 6-(1-fluoro-1-methyl-ethyl)-N4-(2,3,5,6-tetrafluorophenyl)-1,3,5-triazine-2,4-diamine5-triazine-2,4-diamine) (chemical name, CAS registration number: 1606999-21-6) (WO 2014 / 064094, WO 2015 / 162164), (5S)-3-(3-fluoro-5-methyl-phenyl)-N-[rel-(3R,5R)-5-(methoxycarbamoyl)tetrahydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamide -5-methyl-phenyl)-N-[rel-(3R,5R)-5-(methoxycarbamoyl)tetrahydrofuran-3-yl]-5-vinyl-4H-isoxazole-5-carboxamid) (chemical name, CAS registration number: 2266292-43-5) (WO 2018 / 228986, WO 2020 / 114934), 6-(1-methylcyclobutyl)-N4-(2,3,5,6-tetrahydrofuran-3-yl)- 6-(1-methylcyclobutyl)-N4-(2,3,5,6-tetrafluorophenyl)-1,3,5-triazine-4,4-diamine (chemical name, CAS registration number: 1607001-97-7) (WO 2014 / 064094, WO 2015 / 162164), AE-F-150944 (code number), F9960 (code number) Code number), IR-6396 (code number), MCPA-thioethyl, NC-656 (code number), SYP-298 (code number), SYP-300 (code number), S-ethyldipropylthiocarbamate (EPTC), S-metolachlor, S-9750 (code number), MSMA (MSMA), HW-02 (code number), S-523 (code number), SL-1201 (code number).

[0048] Plant growth regulators: 1-naphthylacetamide, 1-methylcyclopropene, 1,3-diphenylurea, 2,3,5-triiodobenzoic acid acid), 2-methyl-4-chlorophenoxybutyric acid (MCPB) [including sodium salt, ethyl ester, etc.], 2-(naphthalene-1-yl)acetamide, 2,6-diisopropylnaphthalene, 3-[(6-chloro-4-phenylquinazolin-2-yl)amino]propane-1-ol, 4-oxo-4-(2-phenylethyl)aminobutyric acid (chemical name, CAS Registry Number: 1083-55-2), 4-chlorophenoxyacetic acid (4-CPA), 5-aminolevulinic acid hydrochloride, methyl 5-(trifluoromethyl)benzo[b]thiophene-2-carboxylate 5-(trifluoromethyl)benzo[b]thiofen-2-carboxylate, AVG (aminoethoxyvinylglycine), n-decyl alcohol (n-decanol), anisiflupurin, aviglycine, ancymidol, abscisic acid, isoprothiolane, inabenfide, indole acetic acid, indole butyric acid, uniconazole, uniconazole-P, Ecolyst, ethychlozate, ethephon, epocholeone, calcium chloride, choline chloridechloride, oxine sulfate, opabactin, kinetin, calcium peroxide, carvone, quinabactin, calcium formate, cloxyfonac, cloxyfonac potassium, cloprop, chlormequat, chlormequat chloride, chlorpropham, choline, cytokinins, oxidized glutathione, cyanamide, sodium cyanate cyanate, cyclanilide, dichlorprop (including salts such as dimethylammonium, potassium, sodium, and choline, or esters such as butotyl ester, 2-ethylhexyl ester, isoctyl ester, and methyl ester), dichlorprop-P (including salts such as sodium, potassium, and dimethylammonium, or 2-ethylhexyl ester), diquat, diquat dibromide, dikegulac, gibberellic acid, gibberellin A4, gibberellin A7, dimethipin, sintophen, jasmone, cis-jasmone, jasmonic acid, and methyl jasmonate jasmonate, streptomycin, calcium sulfur (calcium polysulfide), daminozide, calcium carbonate (calciumcarbonate, thidiazuron, decan-1-ol, triacontanol, triapenthenol, trinexapac-ethyl, tribufos, paclobutrazol, paraffin, bispyribac-sodium, hymexazol, butralin, fluthiacet-methyl, pyraflufen-ethyl ethyl, flumetralin, flurprimidol, flurenol, pronitridine, prohydrojasmon, prohexadione calcium, heptamaloxyloglucan, 6-benzylaminopurine, pendimethalin, forchlorfenuron, formononetin, maleic hydrazide, mepiquat chloride, mefluidide, lipochitooligosaccharides (e.g., lipochitooligosaccharides SP104), calcium sulfate.

[0049] Next, known safeners that may be mixed or used in combination are exemplified below, but are not limited to these examples.

[0050] Safener compounds: Isoxadifen, isoxadifen-ethyl, oxabetrinil, octane-1,8-diamine, cloquintocet, cloquintcet-mexyl, dietholate, cyometrinil, dichlormid, dicyclonone, cyprosulfamide, daimuron, naphthalic anhydride Anhydride, fenchlorazole, fenchlorazole-O-ethyl, fenclorim, furilazole, fluxofenim, flurazole, benoxacor, metcamifen, mephenate, mefenpyr, mefenpi Mefenpyr-ethyl, mefenpyr-diethyl, lower alkyl substituted benzoic acids, 2,2-dichloro-N-(1,3-dioxolan-2-ylmethyl)-N-(2-propenyl)acetamide (PPG-1292), 2-dichloromethyl-2-methyl-1,3-dioxane (MG-191), 3-dichloroacetyl-2,2,5-trimethyl-1,3-oxazolidine (R-29148), 4-dichloroacetyl-1-oxa-4-azaspiro[4.5] Decane (AD-67), 4-carboxy-3,4-dihydro-2H-1-benzopyran-4-acetic acid (CL-304415, code number), MON4660 (code number), metcamifen, N1,N2-diallyl-N2-dichloroacetylglycinamide (DKA-24, code number), 1-bromo-4-[(chloromethyl)sulfonyl]benzene (CSB), 2-propenyl 1-oxa-4-azaspiro[4,5]decane-4-carbodithioate (MG-838, code number), 3-(dichloroacetyl)-2,2-dimethyl-1,3-oxazolidine (R-28725, code number), R-29148 (code number), 1-(dichloroacetyl)azepane (TI-35, code number).

[0051] Next, known biopesticides that may be mixed or used in combination are exemplified below, but are not limited to these examples.

[0052] Biopesticides: Haplothrips brevitubus, Franklinothrips vespiformis, Diglyphus isaea, Encarsia formosa, Amblyseius cucumeris, Pseudaphycus malinus, Amblyseius womersleyi, Aphidius colemani, Eretmocerus eremicus, Aphidoletes aphidimyza, Amblyseius swirskii, Orius strigiformes strigicollis, Phytoseiulus persimilis, Amblyseius degenerans, Phytoseiulus persimilis, Orius sauteri, Dacnusa sibirica, Amblyseius californicus, Chrysoperla nipponensis, and Anicetus beneficus.

[0053] Next, known agricultural materials that may be mixed or used in combination are exemplified below, but are not limited to these examples.

[0054] Agricultural supplies: Ethylene, hypochlorous acid water (limited to that obtained by electrolyzing hydrochloric acid or potassium chloride aqueous solution), baking soda, vinegar, humus, humic acid, fulvic acid, seaweed extract, polysaccharides, amino acids, microbial materials, functional ingredients derived from plants and animals, microbial metabolites, microbial activating materials, soil spreading agents, soil permeability adjusting materials, soil water retention materials, etc., and biostimulants.

[0055] Next, known agricultural fertilizer components that may be mixed or used in combination are exemplified below, but are not limited to these examples.

[0056] Fertilizers include inorganic fertilizers and organic fertilizers, and examples include ammonium chloride, ammonium sulfate, ammonium nitrate, ammonium dihydrogen phosphate, ammonium urea nitrate, urea, lime nitrogen, potassium nitrate, calcium superphosphate, calcium triple superphosphate, potassium dihydrogen phosphate, potassium chloride, potassium sulfate, potassium carbonate, potassium silicate, oil cake, fish meal, rice bran, bat guano, and fermented chicken manure.

[0057] Furthermore, the soil-borne plant disease control agent of the present invention can also be used for plants that have acquired characteristics such as pest resistance, disease resistance, herbicide resistance, etc. through new breeding techniques such as genetic recombination and genome editing, artificial crossbreeding, etc.

[0058] In this way, by using live cells of a Bacillus bacterial strain, such as a Bacillus G4L1 strain, a Bacillus zimonensis G5S1 strain, and / or a Fictibacillus solisarci G5L2 strain, or a culture containing said live cells, as an active ingredient, it is possible to provide a drug that is significantly effective in controlling soil-borne plant diseases such as vegetable bacterial wilt and that can be used safely in actual crop production sites.

[0059] Hereinafter, examples of the present invention will be described, but the present invention is not limited to these examples, and various modifications are possible within the technical concept of the present invention. [Example]

[0060] (Test to confirm effectiveness in preventing bacterial wilt of tomato) Tomato plants (cultivar: Ponderosa) were grown in pots (9 cm x 9 cm) filled with horticultural soil until they reached the four-compound leaf stage. Then, 50 ml of Bacillus sp. G4L1, Bacillus zimonensis G5S1, or Fictibacillus solisarci G5L2 suspensions were hand-sprayed onto the stems and leaves, and the plants were stored in a greenhouse at 35°C for 7 days. Then, 50 ml of the bacterial wilt pathogen suspension was inoculated via bottom watering. For comparison, a control group was also treated with only the bacterial wilt pathogen suspension via bottom watering. The G4L1, G5S1, and G5L2 strain suspensions were cultured in NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.5%, pH 7.0) at 30°C, 200 rpm for 24 hours with shaking, then centrifuged (4500 x g, 15 minutes), suspended in sterilized water, and centrifuged twice for cell collection. The absorbance at 600 nm (OD 600 The tomato bacterial wilt bacteria suspension was prepared by shaking in YP medium (0.5% yeast, 1.0% peptone, pH 6.8-7.2) for 24 hours, then centrifuged to collect the bacteria, which were then diluted with distilled water and measured at 600 nm absorbance (OD 600 ) is 0.02(9.2×10 6 The culture was prepared so that the concentration of the culture medium was 1000 cfu / ml.

[0061] Twenty days after inoculation with the tomato bacterial wilt fungus, the severity of tomato bacterial wilt was indexed according to the following criteria, and the disease severity and control value were calculated. Photographs were also taken of five plants each. The results are shown in Figures 1 and 2, which are used as drawing photographs.

[0062] <Disease index> 0: No disease 1: Some leaflets are wilted 2: Less than half of the compound leaves are wilted 3: More than half of the compound leaves are wilted 4: Withering

[0063] <Disease severity and control value> Severity of disease = Σ (disease index × number of affected strains) / (number of investigated strains × 4) × 100 Control value = 100 - (disease incidence in treated area / disease incidence in untreated area) x 100

[0064] The test results are shown in Table 2 below and Figure 1 (a photograph substituted for a drawing). In the untreated plot, tomato bacterial wilt developed on two-thirds of the leaves, whereas the G4L1 strain-treated, G5S1 strain-treated, and G5L2 strain-treated plots showed a significant decrease in the rate of diseased leaves and a significant decrease in the average disease severity. The control titers were also high, demonstrating that the strains exert a high control effect against tomato bacterial wilt. Figure 2 is a photograph substituted for a drawing showing the results for the untreated plot.

[0065] [Table 2] [Example]

[0066] (Comparative test of the effectiveness of existing Bacillus agents in controlling bacterial wilt of tomato) Tomato plants (cv. Ponderosa) were grown in pots (9 cm × 9 cm) filled with horticultural soil until the four-leaf stage. They were treated with 50 ml of hand sprays of Bacillus sp. G4L1, Bacillus zimonensis G5S1, or Fictibacillus solisarci G5L2 strain suspensions, as well as the conventional Bacillus formulations Impression Clear (registered trademark, manufactured by SDS Biotech), Agrocare Wettable Powder (registered trademark, manufactured by Nippon Soda Co., Ltd.), and Ecoshot (registered trademark, manufactured by Kumiai Chemical Industry Co., Ltd.), and the antibiotic Validacin Liquid 5 (registered trademark, manufactured by Sumitomo Chemical Co., Ltd.). The plants were then inoculated with 50 ml of the bacterial wilt pathogen suspension via submerged infusion. For comparison, a control group was also treated with the bacterial wilt pathogen suspension alone via submerged infusion. The G4L1, G5S1, and G5L2 strain suspensions were cultured in NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.5%, pH 7.0) at 30°C, 200 rpm for 24 hours with shaking, then centrifuged (4500 x g, 15 minutes), suspended in sterilized water, and centrifuged twice for cell collection. The absorbance at 600 nm (OD600 As shown in Table 3 below, the viable cell count in the suspension was 2.3 x 10 for the G4L1 strain suspension. 7 cfu / ml, G5S1 strain suspension was 1.6 × 10 8 cfu / ml, G5L2 strain suspension was 1.4 × 10 7 The bacterial suspension of tomato bacterial wilt was cultured in YP medium (0.5% yeast, 1.0% peptone, pH 6.8-7.2) for 24 hours with shaking, then centrifuged and diluted with distilled water. The absorbance at 600 nm (OD 600 ) is 0.02(9.2×10 6 The viable cell count of the comparative agent was adjusted to 1.6 × 10 cfu / ml. 8 cfu / ml), Agrocare wettable powder (1.6 x 10 8 cfu / ml), EcoShot (1.6 x 10 8 cfu / ml), and an 800-fold diluted solution of Validacin Solution 5 was used in the test.

[0067] Then, 11 days after inoculation with the tomato bacterial wilt fungus, the degree of tomato bacterial wilt disease was investigated using the disease index of Example 1, and the disease degree and control value were calculated.

[0068] The active ingredients of the above-mentioned commercially available control agents are as follows: (A) Impression Clear: Bacillus amyloliquefaciens AT-332 (product of SDS Biotech) (B) Agrocare Wettable Powder: Bacillus subtilis HAI-0404 (Nippon Soda Co., Ltd. product) (C) Ecoshot: Bacillus subtilis D747 strain (Kumiai Chemical Industry Co., Ltd.) (D) Validacin Liquid 5: Validamycin A (Sumitomo Chemical Co., Ltd.)

[0069] The test results are shown in Table 3 below. Compared to the existing bacillus preparations Impression Clear, Agrocare wettable powder, and Ecoshot treatments, the Bacillus sp. G4L1 strain suspension, Bacillus timonensis G5S1 strain suspension, and Fictibacillus solisarci G5L2 strain suspension treatments showed clearly superior control effects against tomato bacterial wilt at equivalent viable cell counts or lower. They also showed clearly superior effects to Validacin Liquid 5, an antibiotic known to be effective against tomato bacterial wilt when sprayed on the stem and leaves.

[0070] [Table 3] [Example]

[0071] (Test to confirm the effectiveness of treating only the aboveground parts of tomatoes against bacterial wilt) To confirm that the bacterial wilt pathogen was indirectly effective against bacterial wilt disease when sprayed onto stems and leaves, tomato plants (Momotaro 8) were grown to the four-compound leaf stage as in Example 1 and then completely covered with aluminum foil to prevent dripping of the bacterial wilt pathogen into the soil (Figure 3). The stems and leaves were then treated with 50 ml of Bacillus sp. G4L1 suspension by hand spray. After complete air drying, the aluminum foil was removed and the plants were stored in a greenhouse at 35°C for 7 days. Subsequently, 50 ml of the tomato bacterial wilt pathogen suspension was inoculated by bottom watering. For comparison, a control group was also treated with only the tomato bacterial wilt pathogen suspension by bottom watering. The G4L1 strain suspension was cultured in NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.5%, pH 7.0) at 30°C, 200 rpm for 24 hours, and then centrifuged (4500 x g, 15 minutes). The cells were then suspended in sterilized water and centrifuged twice for collection. The absorbance at 600 nm (OD 600 The tomato bacterial wilt bacteria suspension was prepared by shaking culture in YP medium for 24 hours, then centrifuged to collect the bacteria, which were then diluted with distilled water and measured at 600 nm absorbance (OD 600 ) is 0.02(9.2×10 6 The culture was prepared so that the concentration of the culture medium was 1000 cfu / ml.

[0072] Then, for each of the 10 treated tomatoes, the degree of bacterial wilt disease was investigated using the disease index of Example 1 13 days after inoculation with the tomato bacterial wilt pathogen, and the disease degree and control value were calculated.

[0073] The test results are shown in Table 4 below. While nearly 60% of the leaves in the untreated plot developed tomato bacterial wilt, the disease severity was clearly reduced in the plot treated with the G4L1 strain, demonstrating that the strain exerts a high indirect control effect against tomato bacterial wilt, a soil-borne disease, when sprayed on stems and leaves.

[0074] [Table 4] [Example]

[0075] (Preparation of spore suspension of Bacillus sp. G4L1) Bacillus sp. G4L1 was inoculated into NA medium and cultured at 30°C for 24 hours. The resulting colonies were cultured in NB medium at 200 rpm at 35°C for 4 days, and then allowed to stand at 4°C for 2 days to promote lysis. Silicone KS-66 (registered trademark: Shin-Etsu Chemical Co., Ltd.) was added as an antifoaming agent each time.

[0076] (Test to confirm efficacy in controlling bacterial soft rot of Chinese cabbage) The Chinese cabbage soft rot fungus (Erwinia carotovora subsp. carotovora) was inoculated and allowed to develop in a glass moist chamber at 28°C for 48 hours. The soft rot-infected field was prepared by spraying and plowing into the field 9 days before transplanting the infected Chinese cabbage. Chinese cabbage (variety: Musou) seedlings were grown for 41 days in 36-well cell trays filled with Yosaku (registered trademark: manufactured by J-Cam Agri Co., Ltd.) soil and transplanted to the soft rot-infected field at intervals of 30 cm between plants and 40 cm between rows. The spore suspension was fluidized and granulated into the Bacillus sp. G4L1 bacterial preparation (5.7 × 10 9The Bacillus sp. G4L1 formulation was not added to the dilution solution, but rather to the wetting agent Kumiai Kumiten (registered trademark, manufactured by Kumiai Chemical Industry Co., Ltd.) at a volume of 1 / 5000. The diluted solution was sprayed twice, 26 and 33 days after transplantation, at a volume of 200 L / 10 a using a gas sprayer. For comparison, a control group was sprayed with a dilution solution of the wetting agent alone, without the Bacillus sp. G4L1 formulation (untreated), and a control group was sprayed with a 500-fold diluted solution of Z Bordeaux Wettable Powder (registered trademark), an inorganic copper agent commonly used to control soft rot in vegetables, to which the same spreading agent had been added.

[0077] 41 days after transplanting, 26 Chinese cabbage plants were harvested from each plant, and the degree of Chinese cabbage soft rot disease was investigated according to the following criteria, and the disease degree and control value were calculated.

[0078] <Disease index> 0: No disease 1: Only some of the outer leaves are affected 2: Disease occurs on some outer leaves and head leaves 3: Most of the head leaves are infected or more severely damaged

[0079] <Disease severity and control value> Severity of disease = Σ (disease index x number of affected plants) x 100 / (number of surveyed plants x 3) Control value = 100 - (disease incidence in treated area / disease incidence in untreated area) x 100

[0080] The results are shown in Table 5. Compared to the disease severity of 23.1 in the untreated area, the Chinese cabbage treated with the G4L1 formulation showed a lower disease severity and a higher control value than Z-Bordeaux WD. Thus, Bacillus sp. G4L1 exerts a high control effect against Chinese cabbage soft rot.

[0081] [Table 5]

[0082] The active ingredients of the above-mentioned commercially available control agents are as follows: (A)Z Bordeaux Wettable Powder: Basic Copper Sulfate (Nippon Nohyaku Co., Ltd. product) [Example]

[0083] (Proliferation confirmation test) The G4L1 strain and commercially available Bacillus disease control agents Impression Clear (registered trademark), Botkiller (registered trademark) wettable powder, and Ecoshot (registered trademark) were inoculated into NA medium (0.5% meat extract, 1.5% peptone, 0.5% sodium chloride, 1.5% agar, pH 7.0) and cultured at 30°C for 24 hours. Single colonies that appeared after 24 hours were picked and cultured in NB medium (0.5% meat extract, 1.5% peptone, 0.5% sodium chloride, 0.5% potassium dihydrogen phosphate, pH 7.0) at 30°C for 24 hours with shaking at 200 rpm. The bacteria were then collected by centrifugation (4500 × g, 15 minutes), suspended in sterilized water, and centrifuged twice. The bacteria were then used. The absorbance at 600 nm (OD ) was then measured. 600 ) was adjusted to 0.1. 100 μl of the prepared bacterial solution was added to an L-shaped test tube filled with 5 ml of NB medium. The culture was performed with shaking at 30°C and 70 rpm in a small shaking culture device, and the absorbance at 600 nm (OD 600 ) was measured to obtain a growth curve.

[0084] The active ingredients of the above-mentioned commercially available control agents are as follows: (A) Impression Clear: Bacillus amyloliquefaciens AT-332 (product of SDS Biotech) (B) Botokilla Hydrate: Bacillus subtilis (Idemitsu Kosan Co., Ltd. product) (C) Ecoshot: Bacillus subtilis D747 (Kumiai Chemical Industry Co., Ltd. product)

[0085] The culture results are shown in Table 6 and Figure 4 (culture curves for the four strains). The G4L1 strain grew at a significantly faster rate than the commercially available Bacillus disease control agents and reached the stationary phase the fastest. Its absorbance was also the highest compared to the other Bacillus disease control agents, demonstrating that the strain grew at a faster rate than conventional strains and could be cultured in a shorter time than conventional strains.

[0086] [Table 6]

[0087] As is clear from the above, the strain of the present invention has a fast growth rate, and as is clear from the absorbance data, it rapidly increases over the 5-8 hour incubation period, and also over the 8-10 hour incubation period. In fact, in the data for the 8-hour incubation period, the absorbance of the present strain is (approximately) twice (or more) the absorbance of the best conventional strain (here, the active ingredient strain in Impression Clear), and a very high absorbance is obtained even after a 10-hour incubation period. In addition, the 600 nm absorbance (OD 600 ) The shortest cultivation time required to obtain a measured absorbance of 3.00 to 3.50 for a conventional Bacillus strain (here, the active ingredient strain in Impression Clear) is 15 hours, whereas this strain reaches this absorbance value after just 10 hours of cultivation, demonstrating its extremely high growth rate.

[0088] Thus, a fast growth rate of the strain used means that the active ingredient can be obtained in a short time, which in turn shortens the production period of the control agent of the present invention. In the production of microbial pesticides, the microbial culture and growth process can be said to be the rate-limiting step of the chemical reaction, so the faster this process is completed, the more quickly the control agent can be produced in a short time. At the same time, this is also beneficial in preventing contamination. Furthermore, there are advantages in the formulation process.

[0089] The present invention can be summarized as follows.

[0090] The present invention aims to provide a novel, non-phytopathogenic strain of bacteria that can be safely used as a biological pesticide and that exhibits stable soil-borne plant disease control effects in actual crop production sites, as well as a soil-borne plant disease control agent and the like that uses the novel strain.

[0091] Furthermore, by using as an active ingredient live cells of novel strains of bacteria belonging to the genus Bacillus or Fictibacillus, which were previously unknown to have a control effect on bacterial plant diseases or soil-borne plant diseases, or a culture containing said live cells, it is possible to provide an agent for controlling soil-borne plant diseases such as an agent for controlling bacterial wilt of vegetables. This agent is also characterized in that it can control bacterial wilt disease and / or bacterial soft rot caused by the soil bacterium Ralstonia solanacearum by foliar (spray) treatment. [Accession number]

[0092] The accession numbers of the microorganisms for which deposit procedures are being carried out in the present invention are shown below. (1) Bacillus sp. G4L1 strain (NITE BP-03204). (2) Bacillus timonensis G5S1 strain (NITE BP-03206). (3) Fictibacillus solisalsi G5L2 strain (NITE BP-03205).

Claims

1. A novel bacterial strain, Bacillus sp. G4L1 strain (NITE BP-03204).

2. A novel bacterial strain, Bacillus timonensis G5S1 strain (NITE BP-03206).

3. A novel bacterial strain, Fictibacillus solisalsi G5L2 strain (NITE BP-03205).

4. The strain according to any one of claims 1 to 3, which is capable of controlling plant diseases by applying it to at least one of plant seeds, roots of plants, stems and leaves of plants, cultivation carriers, nutrient solutions, and soil.

5. A plant disturbance control agent comprising one or more strains or cultures of said strains according to any one of claims 1 to 3 as an active ingredient.

6. 6. The agent according to claim 5, which can control plant diseases by applying it to at least one of plant seeds, roots of plants, stems and leaves of plants, cultivation carriers, nutrient solutions, and soil.

7. 7. The agent according to claim 5 or 6, which is capable of controlling at least one disease of vegetables, fruit trees, rice, and wheat.

8. The agent according to any one of claims 5 to 7, which is capable of controlling diseases of solanaceous plants and / or cruciferous plants.

9. The agent according to any one of claims 6 to 8, which is capable of controlling soil-borne plant diseases by applying it to the stems and leaves of a plant.

10. The agent according to claim 9, which is capable of controlling bacterial wilt and / or soft rot of vegetables.

11. A method for controlling plant diseases, comprising the step of contacting or mixing viable cells of one or more strains according to any one of claims 1 to 3, or a culture containing said viable cells, with or with plant seeds, roots or above-ground parts of plants, cultivation carriers, nutrient solution, or soil.

12. The method according to claim 11, characterized in that it controls at least one disease of vegetables, fruit trees, rice, and wheat.

13. 13. The method according to claim 11 or 12, characterized in that it is used to control diseases of solanaceous plants and / or cruciferous plants.

14. The method according to any one of claims 11 to 13, wherein the method is applied to the stems and leaves of a plant body to control soil-borne plant diseases.

15. The method according to any one of claims 11 to 14, characterized in that it controls vegetable bacterial wilt and / or vegetable soft rot.

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