KR20240001924A - Bacillus amyloliquefaciens SG4-1 strain and uses thereof - Google Patents

Abstract

The present invention relates to the strain of Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) for controlling phytopathogenic fungi. The strain of the present invention is directed against the pathogen ( Boryosphaeria dothidea ) that causes stone fruit resinosis. As an antagonistic microorganism, it has the effect of directly inhibiting the growth of the pathogen, thereby suppressing the occurrence of resinosis in stone fruits (peaches, plums, etc.), which is a plant disease caused by the pathogen, and thus the strain and its The culture medium can be useful as a microbial agent for controlling plant diseases or plant pathogenic fungi.

Description

Bacillus amyloliquefaciens SG4-1 strain and uses thereof {Bacillus amyloliquefaciens SG4-1 strain and uses thereof}

The present invention relates to the strain Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) for controlling phytopathogenic fungi, and also to the use of the strain, the culture medium of the strain, or a mixture thereof as an active ingredient. It relates to a microbial preparation containing the same and a method for controlling plant diseases or plant pathogenic fungi using the same.

In recent years, when cultivating stone fruits such as peaches and plums, the importance of pesticide-free cultivation methods, including organic farming methods, has been increasingly highlighted. However, in the case of current peaches, if pesticides are not used due to damage from pests and birds, there will be a significant decrease in harvest.

In general, when cultivating peach and other stone fruit trees, resin disease often occurs. Resin disease is a general term for symptoms in which resin comes out from the trunk. If resin disease is severe, the tree's viability is weakened and it can die. do. Peach resin disease usually occurs during the rainy season from May to August. The causes are generally excessive nitrogen, poor drainage, excessive dryness, frostbite, scorching, and peach glass moth damage. It is caused by pathogens invading areas weakened by various factors, such as non-infectious ones such as warts, warts, and tuberculosis, and infectious pathogens such as warts, blight, etc., and the tree's resistance to various pathogens appears.

To prevent this type of resin disease, thorough drainage, proper irrigation to prevent drying out, even arrangement of branches so that direct sunlight does not hit the tree trunk, and appropriate control to prevent damage from pests and diseases are carried out. More effectively, the trees are drawn out. Apply chemicals such as whitewash, concentrated lime solution, and water-based paint to remove them during the growing season.

Even if pesticides are sprayed to prevent and treat resin disease, it takes a lot of time to spread it evenly because it is done by machinery and people, so resin disease occurs in areas where pesticides were not sprayed, making it difficult to be effective and treat. In some cases, cutting off branches or, in more severe cases, uprooting the entire tree can result in economic loss.

Due to the above-mentioned problems, various pesticides have been developed to treat and prevent various diseases from diseases and pests when cultivating stone fruits. However, looking at the conventional technology, it is mostly used to prevent resin disease before it occurs or to promote the growth of trees, and to prevent further occurrence after resin disease of stone fruits occurs and to treat resin disease that has already occurred. has the problem of being difficult.

Meanwhile, biological pesticides are pesticides made using active substances produced by plants, insects, and microorganisms that exist in nature through their cultures or extracts. Compared to conventional chemical pesticides, these biological pesticides have less toxicity and resistance to pests and diseases, and are safe for the environment, so there is a lot of interest in developed countries to replace and supplement chemical pesticides. Consumer interest in environmental conservation and food safety is increasing in Korea, and with the implementation of the Positive List System (PLS), much research is being focused on the development of biological pesticides that can replace chemical pesticides. there is. In particular, the demand for agricultural products produced from natural materials such as organic matter is increasing due to the avoidance of the use of chemical materials such as pesticides and chemical pesticides, and the use of microbial preparations that are beneficial to the ecosystem chain and the environment is an alternative to the use of chemical materials. Demand is increasing.

Republic of Korea Patent No. 10-2248585 Republic of Korea Patent Publication No. 10-2014-0034660

The purpose of the present invention is to provide a novel strain that has an antagonistic effect on phytopathogenic fungi that cause stone fruit resinosis.

Another object of the present invention is to provide a microbial agent for controlling plant diseases containing the strain, the culture medium of the strain, or a mixture thereof as an active ingredient.

Another object of the present invention is to provide an antifungal composition against plant pathogenic fungi containing the above strain, a culture medium of the above strain, or a mixture thereof as an active ingredient.

Another object of the present invention is to use at least one selected from the group consisting of the strain, a culture medium of the strain, a microbial preparation containing the strain, and a microbial preparation containing the culture medium of the strain, to a plant, an area around the plant, or these The goal is to provide a plant disease control method that includes treatment steps.

Another object of the present invention is to use at least one selected from the group consisting of the strain, a culture medium of the strain, a microbial preparation containing the strain, and a microbial preparation containing the culture medium of the strain, to a plant, an area around the plant, or these To provide a method for controlling phytopathogenic fungi, including the step of treating all of them.

In order to achieve the above purpose,

The present invention provides a novel microorganism, Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) strain, which has an antagonistic effect on phytopathogenic fungi.

In one embodiment of the present invention, the strain may include a 16S rRNA gene having the base sequence shown in SEQ ID NO: 1.

In one embodiment of the present invention, the strain may have antifungal activity against the pathogen of stone fruit resinous disease.

In one embodiment of the present invention, the stone fruit may be one or more types selected from the group consisting of peaches, plums, apricots, plums, cherries, and jujubes.

In one embodiment of the present invention, the strain may have antagonistic activity against Botryosphaeria dothidea .

Additionally, the present invention provides a microbial agent for controlling plant diseases comprising the above strain, a culture medium of the above strain, or a mixture thereof as an active ingredient.

Furthermore, the present invention provides an antifungal composition against Botryosphaeria dothidea comprising the above strain, a culture medium of the above strain, or a mixture thereof as an active ingredient.

Furthermore, the present invention relates to at least one selected from the group consisting of the strain, a culture medium of the strain, a microbial preparation containing the strain, and a microbial preparation containing the culture medium of the strain, to a plant, an area surrounding the plant, or all of them. Provides a method for controlling plant diseases or plant pathogenic fungi comprising the step of treating.

In one embodiment of the present invention, the plant disease may be stone fruit resinosis.

In one embodiment of the present invention, the plant may be a stone fruit.

In one embodiment of the present invention, the phytopathogenic fungus may be Botryosphaeria dothidea .

In one embodiment of the present invention, the treatment step may be a step of treating any one or more of the strain, the culture medium of the strain, and the microbial agent by immersion, irrigation, or spraying.

The Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) strain of the present invention is an antagonistic microorganism against the pathogen ( Boryosphaeria dothidea ) that causes stone fruit resinosis, and contains auxin, siderophore, cellulase, etc. It has a productivity of and has the effect of directly inhibiting the growth of the pathogen, thereby suppressing the occurrence of resinosis in stone fruits (peaches, plums, etc.), a plant disease caused by the pathogen.

Figure 1 is a graph showing the effect of inhibiting mycelial growth of resinous pathogens of active candidate microorganisms against stone fruit resinosis.
Figure 2 shows the results of identification of antagonistic microorganisms against selected stone fruit resinous diseases.
Figure 3 is a photograph of the visual form of peach resinous lesions in the early (A), middle (B), and late (C) stages of peach resinous disease development as a standard for investigation of peach resinous disease.
Figure 4 is a photograph of the visual form of the lesions in the early (A), middle (B), and late stages (C) of the development of plumu dizziness as a standard for the investigation of plumu dizziness.

Hereinafter, the present invention will be described in detail through embodiments of the present invention with reference to the attached drawings. However, the following embodiments are provided as examples of the present invention, and if it is judged that a detailed description of a technology or configuration well known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description may be omitted. , the present invention is not limited thereby. The present invention is capable of various modifications and applications within the description of the claims described below and the scope of equivalents interpreted therefrom.

In addition, the terminology used in this specification is a term used to appropriately express preferred embodiments of the present invention, and may vary depending on the intention of the user or operator or the customs of the field to which the present invention belongs. Therefore, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part is said to “include” a certain element, this means that it may further include other elements rather than excluding other elements, unless specifically stated to the contrary.

Throughout this specification, '%' used to indicate the concentration of a specific substance means (w/w) % for solid/solid, (w/v) % for solid/liquid, and Liquid/liquid is (v/v) %.

In one aspect, the present invention relates to a novel microorganism Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) strain, which has an antagonistic effect on phytopathogenic fungi.

In one embodiment of the present invention, the strain may include a 16S rRNA gene having the base sequence shown in SEQ ID NO: 1.

In one embodiment of the present invention, the strain may be a strain derived from a ripened agricultural product, and the ripened agricultural product may be a product of natural fermentation and ripening of peaches of low marketability collected in a barrel and placed at room temperature, but is not limited thereto.

In one embodiment of the present invention, the strain may have antifungal activity against the pathogen of stone fruit resinous disease. The pathogen of stone fruit resinosis may be Botryosphaeria dothidea , and the strain of the present invention may have an antagonistic effect against Botryosphaeria dothidea.

In the present invention, stone fruits may include all fruits that have seeds in the ovary but form a hard core in the middle, such as peaches, plums, apricots, plums, cherries, and jujubes, and whose seed coats form the pulp, and stone fruit resin. Any stone fruit that can cause symptoms may be included. Preferably it may be a peach or a plum.

In the present invention, "stone fruit resin disease", also called "stone fruit resin disease", refers to a symptom in which resin, a sticky viscous fluid, is secreted from the main stem or main branch of stone fruit. At first, the resin is transparent and jelly-like. As it is secreted, it gradually turns dark brown and later hardens to become black brown.

The strain of the present invention has the effect of directly inhibiting the mycelial growth of the pathogen ( Boryosphaeria dothidea ) that causes stone fruit resinosis, and increases the productivity of siderophore and cellulase, which are known as components that interfere with the growth of plant pathogens. and auxin (indole-3-acetic acid, IAA). Specifically, it can inhibit spore germination by producing siderophore to form a complex with iron ions and interfere with the absorption of iron by plant pathogenic fungi, and by producing cellulase, it can decompose the cell wall fiber of plant pathogenic fungi, through which stone fruit It may exhibit direct antibacterial activity that inhibits the growth of bacteria that cause resinous disease. Additionally, the strain of the present invention may possess the antibiotics Bacilysin ( bac D), Iturin A ( itu A), and Surfactin ( srf A) as secondary metabolite genes.

When cultivating for mass production of the strain of the present invention, the culture scale may be 10% to 30% by volume of culture solution based on the volume of the culture vessel, and the culture time may be 20 to 82 hours, and should not exceed 25% of the volume of the culture vessel. It may be preferable that the incubation time is at least 48 hours. If it is outside the above range, the mycelial growth inhibition effect and auxin productivity against microbial stone fruit resinous pathogens may be reduced.

In addition, carbon sources during cultivation for mass production of the strain of the present invention include glucose, maltose, galactose, sucrose, arabinose, molasses, and One or more carbon sources selected from the group consisting of corn starch can be used. Preferably, one or more carbon sources selected from the group consisting of arabinose and molasses can be used. When arabinose and molasses are used as carbon sources, the mycelial growth inhibition effect and auxin productivity against stone fruit resin disease pathogens can be improved, and arabinose and molasses can be used as carbon sources. It may be best to use North.

In addition, nitrogen sources during cultivation for mass production of the strain of the present invention include ammonium sulfate, peptone, potassium nitrate, ammonium dihydrogen phosphate, and ammonium dihydrogen phosphate. One or more nitrogen sources selected from the group consisting of (diammonium hydrogen phosphate) can be used. Preferably, at least one nitrogen source selected from the group consisting of ammonium sulfate and peptone can be used. When ammonium sulfate and peptone are used as nitrogen sources, the mycelial growth inhibition effect and auxin productivity against stone fruit resin disease pathogens This can be improved, and it may be most preferable to use ammonium sulfate.

In one aspect, the present invention provides a microbial agent for controlling plant diseases comprising the Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) strain, a culture medium of the strain, or a mixture thereof as an active ingredient. to provide.

In the microbial agent for controlling plant diseases of the present invention, the plant disease may be stone fruit resinosis.

In the microbial agent for controlling plant diseases of the present invention, the plant may be a stone fruit.

In this specification, “culture medium” refers to the result obtained by inoculating a strain into a medium and culturing it for a certain period of time, including the culture medium itself obtained by culturing the strain according to the present invention in a suitable liquid medium, and the filtrate (filtration) obtained by removing the strain by filtering or centrifuging the culture medium. liquid or centrifuged supernatant), cell lysate obtained by sonicating the culture solution or treating the culture solution with lysozyme, the culture solution, filtrate, concentrate of the cell lysate, etc., etc. It is not limited. In addition, the probiotic or antibacterial composition may contain a culture such as a solid medium culture or a dried product or extract of the culture medium, but is not limited thereto, and further contains a suitable excipient or carrier (if the recessive gene is heterozygous). It may additionally include existing objects).

The microbial preparation of the present invention can be prepared in the form of a liquid fertilizer, and can be used in the form of powder by adding an extender, or can be formulated and granulated. However, there is no particular limitation to the formulation.

The microbial preparation of the present invention may further include any other ingredients used for treating plants in the art to the Bacillus amyloliquefaciens SG4-1 strain, a culture medium of the strain, or a mixture thereof. For example, it may further include one or more selected from the group consisting of plant growth regulators, plant growth stimulants, plant nutrients, herbicides, fungicides, pesticides, and fertilizers. However, it is not limited to this.

The plant growth regulator, plant growth stimulant or plant nutrient may be a chemical or another strain of bacteria or any combination thereof. The pesticide may be Anychung, Smithion, Dimechron, Horimat, etc., and preferably, the microbial preparation may further include one or more of Anychung and Smithion.

The plant growth regulators may include other beneficial microorganisms, microbial active substances and organic fertilizers. In addition, chemical fertilizers may be included. However, it is not limited to this.

In one aspect, the present invention provides Bacillus amyloliquefaciens SG4-1 (accession number: KACC 92428P) strain, a culture medium of the strain, or a mixture thereof as an active ingredient. Provides an antifungal composition against Botryosphaeria dothidea ).

In the antifungal composition of the present invention, the description of the strain and culture medium is as above.

In one aspect, the present invention provides a Bacillus amyloliquefaciens SG4-1 (Accession Number: KACC 92428P) strain, a culture medium of the strain, a microbial preparation containing the strain, and a culture medium of the strain. A method for controlling plant diseases is provided, including the step of treating plants, areas around plants, or both with at least one selected from the group consisting of microbial agents.

In one aspect, the present invention provides a Bacillus amyloliquefaciens SG4-1 (Accession Number: KACC 92428P) strain, a culture medium of the strain, a microbial preparation containing the strain, and a culture medium of the strain. A method for controlling phytopathogenic fungi is provided, which includes treating the plant, the area around the plant, or both with at least one selected from the group consisting of microbial preparations.

In the method for controlling plant diseases or controlling plant pathogenic fungi of the present invention, the plant may be a stone fruit.

In one embodiment of the present invention, the plant disease may be stone fruit resinosis and may be caused by Botryosphaeria dothidea .

In one embodiment of the present invention, the phytopathogenic fungus may be a phytopathogenic fungus that causes stone fruit resinosis, and preferably, it may be Botryosphaeria dothidea .

In the method for controlling plant diseases or phytopathogenic fungi of the present invention, the treating step includes treating at least one of the strain, the culture medium of the strain, and the microbial agent by immersing, drenching, or spraying. It can be.

In the method for controlling plant diseases or controlling plant pathogenic fungi of the present invention, a step of removing resinous lesions generated on plants may be additionally performed prior to the treatment step.

In the case of the immersion method, the culture solution and preparation can be irrigated into the soil around the plant or seeds can be immersed in the culture solution and microbial preparation to inoculate. However, the vaccination method is not limited to the above.

The method for controlling plant diseases or controlling phytopathogenic fungi of the present invention includes applying the microbial preparation at any time during the life cycle of the plant, during one or more stages of the life cycle of the plant, or at regular intervals in the life cycle of the plant. , or involves continuous application throughout the life of the plant. Therefore, microbial preparations can be applied as needed. For example, the microbial agent can be applied to the plant during growth, before and/or during flowering, and/or before and/or during seed development. In one example, the microbial agent may be applied before, during and/or immediately after the plants are transplanted from one location to another location, such as from a greenhouse or hotbed to the field. In another example, each microbial agent can be applied to the plant multiple times at desired intervals.

In the method for controlling plant diseases or the method for controlling plant pathogenic fungi of the present invention, with respect to the soil around the plant, the surrounding area to which the present invention is applied means the range in which the microbial agent of the present invention can exert its effect. This includes both above-ground and underground areas. For example, it may be an area within about 2 m, within about 1 m, within about 70 cm, within about 50 cm, within about 25 cm, within about 10 cm, or within about 5 cm around the plant, plant part, or fruit. However, it is not limited to the above area.

In the method for controlling plant diseases or controlling plant pathogenic fungi of the present invention, the part of the plant to which the present invention is applied is not limited. Therefore, it includes not only the whole plant but also parts of the plant, such as seeds, shoots, stems, roots, leaves, and fruits, as well as the whole of them and parts thereof.

In the method for controlling the plant disease or the method for controlling plant pathogenic fungi of the present invention, the strain, the culture medium of the strain, the microbial preparation containing the strain, or the microbial preparation containing the culture medium of the strain, along with Anichung, Smithion, It may be treated in combination with pesticides such as Dimechron and Horimat, preferably with one or more species selected from the group consisting of Anichung and Smithion, and more preferably, the microbial preparation of the present invention. When processing several times, the treatment timing of annichophytes and smicthion may be varied and mixed with microbial preparations.

Hereinafter, the present invention will be described in more detail through examples. However, these are only for explaining the present invention in more detail and do not limit the scope of the present invention to the examples.

Example 1. Isolation of active microorganisms against stone fruit resinosis

1-1. Investigation of auxin productivity of candidate microorganisms

In the present invention, in order to select antagonistic microorganisms against stone fruit (peach, plum) resinosis, antagonistic microorganisms against stone fruit resinosis were selected through auxin productivity analysis of soil-derived candidate microorganisms (8 species, Table 1).

1 g of sample collected from soil and compost was suspended in 9 ml of sterile saline solution (0.85% NaCl), then the supernatant was gradually diluted (10-4 to 10-6 times) and placed on LB agar (Difco, USA) medium. 100 ul of the sample dilution solution was smeared and incubated at 30°C, and each independent colony formed was separated based on shape, color, and gloss. The isolated independent colonies were inoculated with 1 platinum using the stroke method on the same plate medium, and the bacteria were cultured at 30°C for 1 day and purified.

Auxin productivity of pure isolated microorganisms was performed by modifying the method of Jeong et al. (2006). The pure isolated strain was inoculated into LB agar (Difco, USA) medium and cultured at 28°C, and the formed single colony was inoculated into LB broth (Difco, USA) medium and cultured with agitation at 28°C for 48 hours. After centrifuging the culture medium at 13,000 rpm for 10 minutes, double the volume of Salkowski's solution (50 ml 35% HClO ₄ + 1 ml 0.5 M FeCl ₃ ·6H ₂ O) was added to the supernatant and reacted for 30 minutes at 25°C in dark conditions, followed by colorimetric analysis. The absorbance was measured at 535 nm using , and the total amount of auxin was quantified using IAA (Indole-3-acetic acid, Sigma-Aldrich Co. Ltd) as a standard for auxin productivity.

The auxin productivity of each pure isolated candidate microorganism is shown in Table 1.

candidate microorganism Auxin content
(mM IAA) Microorganism name source DS-1 T1 soil 0.010±0.004 DW'-1 T2 soil 0.043±0.010 DW'-2 T3 soil 0.040±0.003 DW6 T4 soil 0.029±0.004 SG4-1 T5 spoiled agricultural products 0.079±0.013 PNL10-2 T6 soil 0.260±0.010 Seo2020-1 T7 compost 0.061±0.011 D3 T8 soil 0.005±0.001

Among each candidate microorganism, the microorganism with the highest auxin productivity was PNL10-2(T6), followed by SG4-1(T5) and Seo2020-1(T7) with auxin productivity of 0.079±0.013 and 0.061±0.011 mM IAA, respectively. It was found that it had a fairly high auxin productivity.

1-2. Peach resinous disease pathogen mycelial growth inhibition test of candidate microorganisms

In the present invention, in order to select microorganisms with antibacterial activity against stone fruit resinosis, the soil-derived candidate microorganisms are targeted through the growth inhibition ability of pathogens known to cause peach resinosis ( Botryosphaeria dothidea ). Microorganisms antagonistic to stone fruit resinous disease were selected. For this purpose, Botryoshaeria dothidea was distributed and the mycelial growth inhibition ability of each candidate microorganism was investigated using the paper disc method.

Specifically, each candidate microorganism was cultured with agitation in LB broth (Difco, USA) medium, spread on LB agar (Difco, USA) medium, and cultured at 28°C for 48 hours. The colonies formed were cultured again in LB broth (Difco, USA). Co., Ltd., USA) medium was inoculated and cultured with agitation at 28°C for 48 hours, and then the culture was used.

The effect of each candidate microorganism on inhibiting mycelial growth of the resinous pathogen ( Botryosphaeria dothidea ) was evaluated using the following calculation formula, and the results are shown in Table 2 and Figure 1.

[formula]

Mycelial growth inhibition rate (%) = ((total distance - mycelial growth distance) ÷ total distance) × 100

candidate microorganism Mycelial growth inhibition rate (%) Microorganism name source Day 2 of vaccination 3rd day of vaccination DS-1 T1 soil 11.5 0.0 DW'-1 T2 soil 0.0 0.0 DW'-2 T3 soil 4.3 0.0 DW6 T4 soil 15.4 19.2 SG4-1 T5 spoiled agricultural products 51.9 51.9 PNL10-2 T6 soil 8.3 0.0 Seo2020-1 T7 compost 46.2 38.5 D3 T8 soil 19.2 7.7

Two days after inoculation on PDA agar medium, there was no mycelial growth inhibition effect in the case of DW'-1(T2), and there was no inhibitory effect on mycelial growth in DS-1(T1), DW'-2(T3) and PNL10-2(T6). ), the mycelial growth inhibition rates of the resinous pathogen were found to be 11.5%, 4.3%, and 8.3%, respectively.

In the case of PNL10-2(T6), which had the highest auxin productivity among the candidate microorganisms, the mycelial growth inhibition rate against resinous pathogens was found to be quite low.

The inhibition rate of mycelial growth of resinous pathogens of each candidate microorganism was highest at 51.9% on the 2nd day of inoculation in the case of SG4-1 (T5), and the effect was found to be maintained on the 3rd day of inoculation (Table 1, Figure 1). ).

In the case of candidate microorganism SG4-1(T5), auxin productivity was also found to be quite high at 0.079±0.013mM IAA. Accordingly, in the present invention, SG4-1(T5) microorganism was finally selected as a microorganism for controlling stone fruit resinosis and used in subsequent experiments.

1-3. Selection and identification of stone fruit resinous antagonistic microorganisms

The SG4-1(T5) strain selected was identified through the auxin productivity and mycelial growth inhibition tests of the resinous pathogen ( Botryosphaeria dothidea ).

First, for taxonomic identification of the isolated and selected microorganisms, 16S rDNA was amplified by PCR, the sequence was compared with the base sequence registered in NCBI, homology was examined, and final identification was made. The 16S rDNA sequence was identified by extracting the dye DNA of the selected microorganism and performing PCR using a PCR primer. As a result, the SG4-1(T5) strain selected as an antagonistic microorganism of stone fruit resin disease in this research project was Bacillus amylolique. It was finally identified as Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) (Figure 2).

Example 2. Efficacy/effect test of active microorganisms on stone fruit resin symptoms

2-1. Test method and treatment

In order to confirm the efficacy and effect of the selected microorganisms against stone fruit resinosis at the cultivation and packaging level, we requested the Kyungpook National University Agricultural Product Quality and Safety Evaluation Research Institute, a public certification agency, to confirm the efficacy and effects of the selected microorganisms against stone fruit resinosis. An orchard where representative stone fruits, such as peaches and plums, are grown was selected and a field application test was conducted.

The selected antagonistic microorganism, Bacillus amyloliquefaciens , was cultured with agitation in LB broth (Difco, USA) medium, spread on LB agar (Difco, USA) medium, and cultured at 28°C for 48 hours. The colonies formed were then cultured again in LB broth ( Difco, USA) medium was inoculated and cultured with agitation at 28°C for 48 hours, and then the culture was diluted 50 times with water and used as a liquid microbial preparation. The physicochemical properties of the microbial preparations are shown in Table 3 below.

Analysis items (unit) Analysis performance note ingredient Total amount of nitrogen (T-N) 0.15% Tested in accordance with fertilizer quality inspection methods and sample collection standards (Rural Development Administration Notice No. 2020-29, 2020.11.25.) Total amount of phosphoric acid (P ₂ O ₅ ) 0.028% Total amount of potassium (K ₂ O) 0.027% Soluble lime (CaO) 0.0057% Water-soluble kaolin (MgO) 0.0026% Water-soluble silicic acid (SiO ₂ ) Not detected Soluble iron (Fe) Not detected Water-soluble copper (Cu) Not detected Water-soluble boron (B ₂ O ₃ ) 0.064% Water-soluble manganese (MnO) Not detected Water-soluble molybdenum (Mo) 0.0011 Harmful ingredients Arsenic (As) Not detected Nickel (Ni) Not detected Chrome (Cr) Not detected Titanium (Ti) Not detected Cadmium (Cd) Not detected Nitrous acid (NO ₂ ) Not detected Sulfurous acid (SO ₂ ) Not detected

For the field application test, the resinous lesions formed on the trunks of peach or plum trees were scraped off or not, and a 50-fold diluted microbial preparation of the selected antagonistic microorganisms was treated around the lesions using a spray. , it was judged that the efficacy was recognized when the treated resinous lesions were inspected and resin no longer came out. In the case where the microbial agent was treated after removing the resinous lesions, the number of additional occurrences of the resinous lesions was counted and determined. If the microbial agent was treated without removing the resinous lesions, the appearance changes in the resinous lesions were investigated. This was used as the standard for judgment.

In addition, stone fruit resinosis can be broadly divided into physical causes, i.e., resinosis caused by frost damage, scorching, and mechanical wounds, and biological causes, i.e., resinosis caused by pest damage. The efficacy and effectiveness of microbial preparations according to these causes were confirmed. A test was conducted to compare and evaluate changes in lesions when the microbial agent of the present invention was used alone or when a microbial agent and pesticides (pesticide products A and S products) were used together. Descriptions of the untreated group, microbial agent treated group, microbial agent treated group, and pesticide treated group are shown in Table 4 below.

treatment area Ineffectiveness test compare test Spray amount Fertilization method and timing standard amount microbial preparation
treatment area standard amount For each of the resin lesion removal area and the resin lesion non-removal area, a microbial agent (50 times the culture medium of the selected antagonistic microorganisms) was sprayed around the resin lesions using a sprayer so that the chemical solution flows. Standard amount (50 times the culture medium of selected antagonistic microorganisms)
sparge Microbial preparations and pesticide treatment equipment standard amount For each of the resin lesion removal area and the resin lesion non-removal area, a microbial preparation (50 times the culture medium of the selected antagonistic microorganisms) + 2 types of pesticides (A product, S product) were sprayed around the resin lesions using a sprayer so that the chemical solution flows. Non-processed area - -

2-2. 1st packaging test on peaches (microorganism treatment group)

The first peach packaging test was conducted at a peach test packaging over 10 years old located in Hakhari, Gasan-myeon, Chilgok-gun, Gyeongsangbuk-do.

The treatment method of this test is to divide the stem area of the peach tree where peach resinous lesions are observed into a treatment group in which the peach resinous lesions are removed (resin lesion removal group) or a treatment group in which the peach resinous lesions are not removed (resin lesion non-removal group). (50 times the culture medium of the selected antagonistic microorganisms) was treated with a sprayer.

First, in order to confirm the efficacy and effect of the microbial agent of the present invention on peach resinous lesions in the resinous lesion removal area, the number of resinous lesions before treatment was investigated, and then the microbial agent was sprayed and treated with the peach resinous lesions removed. The number of peach resinous lesions in each test group was investigated on days 0, 11, 17, and 67. The number of lesions before treatment in each treatment group varied for each treatment group as shown in Table 5, and the number of lesions in the total treatment group was found to be 69. After examining the number of lesions in each treatment group, the lesions were immediately removed and then treated with a microbial agent. After 11 days of treatment, there were 12 lesions (83% reduction), on the 17th day, 9 lesions (87% reduction), and on the final 67th day, 8 lesions (83% reduction). 88% reduction) was investigated. In other words, the number of resinous lesions was significantly reduced by treatment with the microbial preparation, and the number of lesions was reduced by 61 (approximately 87% reduction) in the final 67 days, showing that the effect was significant.

treatment area
(Less lesion removal tool) Number of digitalis lesions (units) Before processing Day 0 of processing Day 11 of processing Day 17 of processing 67th day of processing KGT-14 19 0 0 0 0 KGT-15 9 0 0 0 0 KGT-16 3 0 2 2 2 KGT-17 2 0 One One 0 KGT-18 5 0 3 2 2 KGT-19 7 0 One 0 3 KGT-20 13 0 2 One One KGT-21 3 0 0 0 0 KGT-22 8 0 3 3 0 Sum 69
(100%) 0 12
(17%) 9 (13%) 8 (12%)

In addition, after examining the peach resinous lesions, the lesions were treated with a microbial agent without removing the lesions, and then the number of lesions in each treatment group was investigated and shown in Table 6. The number of resinous lesions in the entire treatment group before treatment was 197, and after 11 days of treatment with the microbial agent, no lesions were observed in the three treatment groups, and the overall decrease was about 68%. After 17 days, no lesions were observed in 9 treatments, and the number of lesions in other treatments also showed a tendency to rapidly decrease, with an overall decrease of more than 85%.

In particular, on the 67th day of microbial agent treatment, peach resinous lesions were not observed in 18 treatment groups, and the overall reduction was more than 96%, which was judged to have a curative effect. In conclusion, when treating peaches with microbial agents, the effect was relatively higher in the treatment group in which resinous lesions were not removed compared to the treatment group in which the resinous lesions were removed.

treatment area
(Different lesions not removed) Number of digitalis lesions (units) Before processing Day 0 of processing Day 11 of processing Day 17 of processing 67th day of processing KGT-1 One One 0 0 0 KGT-2 15 15 3 2 One KGT-3 3 3 0 0 0 KGT-4 3 3 3 2 One KGT-5 2 2 2 0 0 KGT-6 10 10 3 2 2 KGT-7 2 2 2 One 0 KGT-8 6 6 One 0 0 KGT-9 20 20 0 0 0 KGT-10 9 9 5 2 0 KGT-11 6 6 2 0 0 KGT-12 5 5 2 One 0 KGT-13 5 5 3 2 0 KGT-23 7 7 4 2 0 KGT-24 23 23 2 0 0 KGT-25 7 7 2 2 0 KGT-26 7 7 3 2 0 KGT-27 13 13 7 0 0 KGT-28 10 10 2 0 0 KGT-29 15 15 3 One 0 KGT-30 9 9 6 5 3 KGT-31 5 5 One One 0 KGT-32 14 14 7 4 One Sum 197
(100%) 197
(100%) 63
(32%) 29
(15%) 8
(4%)

Meanwhile, the natural occurrence trend of peach resinous disease by cultivation period in the untreated test field used in this first test is shown in Table 7. On May 21st, at the beginning of peach cultivation, 977 peach resinous lesions were observed, and the number continued to increase somewhat, with 1,057 lesions observed on June 1st and 1,068 lesions on June 7th. The number continued to increase toward the middle of cultivation, and on July 27, 1,191 resinous lesions were observed. In conclusion, in the case of the Chilgok peach test field, the daily natural increase/decrease ratio of peach resinous disease during the cultivation period was found to increase by 3.19 numbers/day.

division Number of lesions by survey date (units) daily nature
increase/decrease ratio
(Number/Day) May 21st June 1st June 7th July 27 gist 910 980 987 1,120 Deputy Governor 67 77 81 71 Sum 977 1,057 1,068 1,191 3.19

2-3. Second packaging test for peaches (microorganism preparations and pesticide treatment equipment)

In this test, it was carried out in a peach field located in Apo-eup, Gimcheon-si, Gyeongsangbuk-do to verify the efficacy, effectiveness, and versatility of the microbial agent of the present invention for peach resinosis caused by wounds caused by borers, among biological causes.

This test packaging was tested by randomly selecting a packaging with severe damage from borers (glass moths, etc.), and the test treatment group was divided into a treatment group in which peach resinous lesions were removed and a treatment group in which peach resinous lesions were not removed, and then the microbial agent of the present invention and the pesticide ( 1st-A product: flubendiamide 20% liquid moisturizer, 2nd-S product: fenitrothion 50% emulsion) were used together for treatment.

First, without removing peach resinous lesions, a mixture of microbial agents and pesticides (1st product A, 2nd product S) was used and the number of lesions in each treatment group was observed. This test was conducted on five treatment groups, and the number of resinous lesions before treatment in each treatment group was as shown in Table 8, and the number of resinous lesions in the entire treatment group before treatment was found to be 255. During the mixed treatment, there were differences for each treatment group, but there was a tendency to decrease. Overall, on the 11th day of treatment, the number of lesions decreased by 69 to 186 (17% decrease) compared to the number of lesions before treatment of 255, and on the 31st day of treatment, the number decreased by 50 to 205. (20% reduction) And on the 64th day of treatment, the number of lesions was observed to be 77 (70% reduction), a decrease of 178.

treatment area
(Different lesions not removed) Number of digitalis lesions (units) Before processing Treatment Day 0 ¹⁾ Day 11 of processing 31st day of processing ²⁾ 64th day of processing KA1-21 27 27 20 17 2 KA1-22 75 75 64 56 30 KA1-23 78 78 42 61 13 KA1-24 42 42 37 30 8 KA1-25 33 33 23 41 24 Sum 255
(100%) 255
(100%) 186
(73%) 205
(80%) 77
(30%) ¹⁾ Pesticide (Product A) mixed treatment, ²⁾ Pesticide (S product) mixed treatment

In addition, after removing the peach resinous lesions, the number of lesions in each treatment group was observed after mixed treatment with microbial agents and pesticides (1st product A, 2nd product S). The number of resinous lesions before treatment in each treatment group was as shown in Table 9, and the number of resinous lesions in the entire treatment group before treatment was found to be 218. On the 11th day of mixed treatment with microbial preparations and pesticides, the number of lesions was 50 (77% decrease), which was 168 less than the number of 218 before treatment, and on the 31st day of treatment, the number was 82, which was 32 more than on the 11th day of treatment, 62% decrease). It increased, but decreased again on the 64th day of treatment, and the number of lesions was observed at 15 (93% reduction).

treatment area
(Less lesion removal tool) Number of digitalis lesions (units) Before processing Treatment Day 0 ¹⁾ Day 11 of processing 31st day of processing ²⁾ 64th day of processing KA1-26 93 0 12 28 0 KA1-27 36 0 7 10 4 KA1-28 31 0 4 One One KA1-29 26 0 19 32 10 KA1-30 32 0 8 11 0 Sum 218
(100%) 0
(0%) 50
(23%) 82
(38%) 15
(7%) ¹⁾ Pesticide (Product A) mixed treatment, ²⁾ Pesticide (S product) mixed treatment

Meanwhile, the natural occurrence trend of peach resinous disease by cultivation period in the untreated test field used in this second test is shown in Table 10. On July 16th, the middle of peach cultivation, 1,262 peach resinous lesions were observed, and the number continued to increase somewhat, with 1,749 peach resinous lesions observed on July 23rd. On August 13th, the second half of cultivation, the number slightly increased from 1,748 to 1,821 on September 2nd, and on October 5th after harvest, the number continued to increase to 1,912. In this survey, peach resinous disease continued to increase from the mid-cultivation period through the late cultivation period to post-harvest. Finally, in the case of Gimcheon packaging, the daily natural increase/decrease ratio of peach resinous disease during the cultivation period was found to increase to 8.02 per day.

division Number of lesions by survey date (units) daily nature
increase/decrease ratio
(Number/Day) July 16th July 23 August 13th September 2nd October 5th gist 1,262 1,749 1,748 1,821 1,912 Deputy Governor 0 0 0 0 0 Sum 1,262 1,749 1,748 1,821 1,912 8.02

2-4. Incidence rate of peach resinous symptoms considering natural increase/decrease changes

Table 11 shows a comparison of the occurrence rates of peach resinous symptoms when considering the natural increase and decrease rate of peach resinous disease and when it did not.

test
packaging
region Processing method Resin symptom occurrence rate (%) daily nature
increase/decrease ratio
(Number/Day) When natural increase/decrease changes are not taken into account When considering natural increase/decrease changes chilgok After removing the lesion
Single-use treatment with microbial agent 11.59 2.82 3.19 Without removing the lesion
Single-use treatment with microbial agent 4.06 1.94 Gimcheon Mixed treatment of microbial agents and pesticides without removing lesions 30.19 10.02 8.02 After removing the lesion, mixed treatment with microbial agent and pesticide 6.88 2.04

First, in the case of Chilgok pavement (1st test), the damage from borers was minimal. In this case, when comparing the incidence of resin symptoms according to whether or not the lesion was removed, the rate of resin symptom occurrence was 11.59% when the lesion was removed and treated with a single microbial preparation without taking into account natural increase or decrease changes. In the case where it was not removed, the rate was 4.06%, so it was better to treat the lesion with a single microbial agent without removing the lesion. When the daily natural increase/decrease rate was taken into account, the incidence rates of resin symptoms were 2.82% and 1.94%, respectively, showing excellent results. In conclusion, in the case of peaches, when damage from borers is small, such as in the Chilgok test field, when treating with only a microbial agent without removing lesions, considering the natural increase or decrease in changes, the incidence of resin symptoms was 1.94%, confirming that the treatment effect was large.

In the case of Gimcheon pavement (2nd test), there is a characteristic of the pavement that there is a lot of damage from borers. When treated with a mixture of microbial agents and pesticides to reduce damage from borers, after removing the lesions, 6.88% of lesions showed signs of resin, which was greater than when they were not removed. This showed a decreasing result. In other words, in the case of fields with significant borer damage, removing lesions when treated with pesticides showed a better effect, and considering natural increase and decrease changes, the resin symptom occurrence rate was 2.04%, which was an excellent result. In conclusion, in the case of the Gimcheon test field, the damage from borers was high, and in this case, after removing the lesions, the mixed treatment with microbial preparations and pesticides was confirmed to have the greatest effect, with a 2.04% rate of resin symptoms taking into account natural increase and decrease changes.

2-5. Primary packaging test for plums (microbiological agent treatment group)

While investigating the Gunwi area in Gyeongbuk on July 2, 2021 to select the field to be used in this test, plum resinosis occurred in an orchard where plum trees over 10 years old were planted, so this test field was selected. In order to confirm the efficacy and effect of the microbial preparation of the present invention on plum resinosis, the plum resinosis lesions were removed and the microbial preparation was treated, and then the number of plum resinosis lesions in each treatment group was observed. This test was conducted on 11 treatment groups. The number of resinous lesions before treatment for each treatment group is shown in Table 12, and the total number of resinous lesions was 59. 12 days after removing the resinous lesions and treating the microbial agent in all 11 treatment groups, 19 resinous lesions (68% reduction) were observed, which is 40 fewer lesions than before treatment. On the 40th day of treatment, 32 lesions were observed. It was found that the number of dogs decreased, resulting in 27 digitalis lesions (54% decrease).

treatment area
(Less lesion removal tool) Number of digitalis lesions (units) Before processing Day 0 of processing Day 12 of processing 40th day of processing KK4-1 4 0 0 0 KK4-2 One 0 0 0 KK4-3 3 0 0 0 KK4-4 2 0 0 0 KK4-5 One 0 One One KK4-6 5 0 0 4 KK4-7 15 0 15 15 KK4-8 5 0 0 0 KK4-9 3 0 One 3 KK4-10 10 0 One 4 KK4-11 10 0 One 0 Sum 59
(100%) 0 19
(32%) 27
(46%)

Meanwhile, the natural change in plum resinosis by cultivation period in the untreated test field used in this first test is shown in Table 13. On July 7th, the middle of plum cultivation, 89 plum resinous lesions were observed, and the number continued to increase somewhat, to 119 on July 14th, and on August 11th, the number increased significantly to 434 lesions. In this investigation, plum resinosis increased significantly from the mid-cultivation period to the late-cultivation period, and it was found that the occurrence was increasing due to damage by borers. The daily natural increase/decrease ratio of this test field was found to be 8.41 per day, and it is judged that the occurrence of resinosis due to borers is continuously increasing.

division Number of lesions by survey date (units) daily nature
increase/decrease ratio
(Number/Day) July 2nd July 14th August 11th gist 89 116 427 Deputy Governor 0 3 7 Sum 89 119 434 8.41

2-6. Secondary field testing on plums (microbiological preparations and/or pesticide treatments)

In this test, it was carried out in a plum cultivation field located in Yeongcheon-si, Gyeongsangbuk-do to verify the efficacy, effectiveness, and versatility of the microbial agent of the present invention against plum resinosis caused by wounds inflicted by pests among biological causes. In order to confirm the efficacy and effect of the microbial preparation for plum resinous disease, the microbial preparation was treated as a single treatment with the lesions removed, and the number of resinous lesions before and after treatment was investigated, and the results are shown in Table 14. The number of resinous lesions in the entire treatment group before treatment was found to be 51, and showed an overall tendency to decrease according to the single-use treatment of the microbial agent.

treatment area
(Single-use treatment for resinous lesion removal tool) Number of digitalis lesions (units) Before processing Day 0 of processing Day 28 of processing 40th day of processing 67th day of processing KY4-1 7 0 One One One KY4-2 12 0 One 2 0 KY4-3 6 0 2 2 One KY4-4 3 0 One 2 2 KY4-5 5 0 3 3 10 KY4-6 3 0 3 2 One KY4-7 9 0 9 4 3 KY4-8 3 0 6 9 9 KY4-9 One 0 0 One 0 KY4-10 2 0 2 2 10 Sum 51
(100%) 0 28
(55%) 28
(55%) 37
(73%)

In addition, in order to confirm the effect of the microbial agent of the present invention on resinosis when mixed with pesticides (Product A: Flubendiamide 20% liquid wettable powder, Product S: Fenitrothion 50% emulsion) to control biological pests, This test was further performed.

To verify the versatility of the microbial agent for peach resinosis, the number of lesions in each treatment group was observed after treating the plum resinous disease lesions with a mixture of microbial agent and pesticide (Product A) without removing the lesions. This test was conducted on five treatments, and the number of resinous lesions before and after treatment for each treatment is shown in Table 15. The number of resinous lesions in all treatment groups before treatment was found to be 141, and there was a difference for each treatment group during mixed treatment, but it showed a tendency to decrease. Overall, there was a tendency to slightly decrease after treatment, and on the 31st day of treatment, 83 lesions were observed, a decrease of approximately 41%.

treatment area
(Mixed treatment for areas where resinous lesions were not removed) Number of digitalis lesions (units) Before processing Treatment Day 0 ¹⁾ Day 12 of processing 20th day of processing 31st day of processing KY4-11 33 33 20 16 10 KY4-12 9 9 7 6 5 KY4-13 20 20 18 18 13 KY4-14 3 3 7 5 3 KY4-15 23 23 20 20 20 KY4-16 5 5 2 2 2 KY4-17 5 5 3 3 3 KY4-18 12 12 8 6 5 KY4-19 16 16 15 15 15 KY4-20 15 15 12 10 7 Sum 141
(100%) 141
(100%) 112
(79%) 101
(72) 83
(59%) ¹⁾ Mixed treatment of microbial agent and pesticide (Product A)

In addition, in order to confirm the efficacy, effectiveness, and versatility of the microbial agent for plum resinosis, the plum resinosis lesions were removed, treated with a mixture of microbial agent and pesticide (S product), and the number of plum resinosis lesions in each treatment group was observed. did. This test was conducted on 10 treatments, and the number of resinous lesions before treatment for each treatment is shown in Table 16. The number of resinous lesions in the entire treatment group before treatment was found to be 113. After 10 days of mixed treatment with microbial agents and pesticides (Product S) in all 10 treatment groups without removing the resinous lesions, 94 resinous lesions were observed, which was 19 less than the number before treatment, and 47 after treatment. It was found that 89 digitalis lesions (21% reduction) occurred in the first day, with a decrease of 24.

treatment area
(Mixed treatment for areas where resinous lesions were not removed) Number of digitalis lesions (units) Before processing Treatment Day 0 ¹⁾ 10th day of processing Day 47 of processing KY5-1 17 17 13 20 KY5-2 18 18 10 6 KY5-3 11 11 10 7 KY5-4 9 9 10 5 KY5-5 12 12 12 15 KY5-6 9 9 9 5 KY5-7 6 6 6 4 KY5-8 9 9 9 9 KY5-9 16 16 12 15 KY5-10 6 6 3 3 Sum 113
(100%) 113
(100%) 94
(83%) 89
(79%)

Meanwhile, the natural occurrence trend of plum resinosis by cultivation period in the untreated test field used in this second test is shown in Table 17. At the beginning of plum cultivation, 264 plum resinous lesions were observed on May 24th and 412 on June 21st. On July 5th, the mid-cultivation period, the number of lesions increased slightly, with 664 lesions observed, and on July 30th, 934 lesions were observed, showing a significant increase.

division Number of lesions by survey date (units) daily nature
increase/decrease ratio
(Number/Day) May 24th June 21st July 5th July 30th gist 260 391 635 934 Deputy Governor 4 21 29 0 Sum 264 412 664 934 10.0

2-7. Prunus Tuberculosis Incidence rate of Tuberculosis symptoms considering natural increase/decrease changes

Table 18 shows a comparison of the incidence rates of plum resinous symptoms when natural increase/decrease changes were taken into account and when it was not.

test
packaging
region Processing method Resin symptom occurrence rate (%) daily nature
increase/decrease ratio
(Number/Day) When natural increase/decrease changes are not taken into account When considering natural increase/decrease changes Gunwi After removing the lesion
Single-use treatment of microbial agent 45.76 6.82 8.41 Yeongcheon Without removing the lesion
Single-use treatment with microbial agent 72.54 5.13 10.0 Mixed treatment of microbial preparations and pesticides (Product A) without removing lesions 58.86 18.40 After removing the lesion, mixed treatment with microbial agent and pesticide (S product) 78.76 15.26

In both Gunwi pavement (first test) and Yeongcheon pavement (second test), the daily natural increase/decrease ratio was found to be large at 8.41 and 10.0 pieces/day, respectively.

The rate of occurrence of resin symptoms in the entire packaging was large when natural changes in increase or decrease were not taken into account, so the treatment effect was judged to be low. However, when taking natural changes in increase or decrease into account, the rate of resin symptoms when treated with a single microbial preparation was 6.82% when lesions were removed and 6.82% when lesions were removed. It was found that when the microbial agent was treated only without removal, it significantly decreased to 5.13%.

In the case of plums, it was found that spraying pesticides in each field had little effect, and it was found that spraying pesticides actually had an effect that hindered the effect of reducing the occurrence of resin symptoms. In conclusion, the incidence of resin symptoms, taking into account natural increase/decrease changes in the treatment of single-use microbial preparations, was 6.82% in Gunwi Pavilion and 5.13% in Yeongcheon Pavement, confirming that the effect was the greatest.

Rural Development Administration National Academy of Agricultural Sciences Microbial Bank (KACC) KACC92428P 20220610

<110> DaewonAgro CO.,Ltd. <120> Bacillus amyloliquefaciens SG4-1 strain and uses it <130> PN2206-313 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1403 <212> RNA <213> Bacillus amyloliquefaciens <400> 1 gcagtcgagc ggacagatgg gagcttgctc cctgatgtta gcggcggacg ggtgagtaac 60 acgtgggtaa cctgcctgta agactgggat aactccggga aaccggggct aataccggat 120 ggttgtctga accgcatggt tcagacataa aaggtggctt cggctaccac ttacagatgg 180 acccgcggcg cattagctag ttggtgaggt aacggctcac caaggcgacg atgcgtagcc 240 gacctgagag ggtgatcggc cacactggga ctgagacacg gcccagactc ctacgggagg 300 cagcagtagg gaatcttccg caatggacga aagtctgacg gagcaacgcc gcgtgagtga 360 tgaaggtttt cggatcgtaa agctctgttg ttagggaaga acaagtgccg ttcaaatagg 420 gcggcacctt gacggtacct aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg 480 taatacgtag gtggcaagcg ttgtccggaa ttattgggcg taaagggctc gcaggcggtt 540 tcttaagtct gatgtgaaag cccccggctc aaccggggag ggtcattgga aactggggaa 600 cttgagtgca gaagaggaga gtggaattcc acgtgtagcg gtgaaatgcg tagagatgtg 660 gaggaacacc agtggcgaag gcgactctct ggtctgtaac tgacgctgag gagcgaaagc 720 gtggggagcg aacaggatta gataccctgg tagtccacgc cgtaaacgat gagtgctaag 780 tgttaggggg tttccgcccc ttagtgctgc agctaacgca ttaagcactc cgcctgggga 840 gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca 900 tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt cttgacatcc tctgacaatc 960 ctagagatag gacgtcccct tcggggggcag agtgacaggt ggtgcatggt tgtcgtcagc 1020 tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aacccttgat cttagttgcc 1080 agcattcagt tgggcactct aaggtgactg ccggtgacaa accggaggaa ggtggggatg 1140 acgtcaaatc atcatgcccc ttatgacctg ggctacacac gtgctacaat ggacagaaca 1200 aagggcagcg aaaccgcgag gttaagccaa tcccacaaat ctgttctcag ttcggatcgc 1260 agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa tcgcggatca gcatgccgcg 1320 gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccacgagagt ttgtaacacc 1380 cgaagtcggt gaggtaacct tta 1403

Claims (14)

Bacillus amyloliquefaciens SG4-1 (Accession number: KACC 92428P) strain, a novel microorganism with antagonistic activity against plant pathogenic fungi. The strain according to claim 1, wherein the strain contains a 16S rRNA gene having the base sequence shown in SEQ ID NO: 1. The strain according to claim 1, wherein the strain has antifungal activity against pathogens of stone fruit resinosis. The strain according to claim 3, wherein the stone fruit is at least one selected from the group consisting of peaches, plums, apricots, plums, cherries, and jujubes. The strain of claim 1, wherein the strain has antagonistic activity against Botryosphaeria dothidea . A microbial preparation for controlling plant diseases comprising the strain according to any one of claims 1 to 5, a culture medium of the strain, or a mixture thereof as an active ingredient. 7. The microbial preparation according to claim 6, wherein the plant disease is stone fruit resinosis. The microbial agent according to claim 6, wherein the plant disease is caused by Botryosphaeria dothidea . An antifungal composition against Botryosphaeria dothidea , comprising the strain according to any one of claims 1 to 5, a culture medium of the strain, or a mixture thereof as an active ingredient. Plants, plants, at least one selected from the group consisting of the strain according to any one of claims 1 to 5, a culture medium of the strain, a microbial preparation containing the strain, and a microbial preparation containing the culture medium of the strain. A method for controlling a plant disease comprising treating the surrounding area, or both. The method of claim 10, wherein the plant is a stone fruit. The method of claim 10, wherein the treating step is treating one or more of the strain, the culture medium of the strain, and the microbial agent by immersing, drenching, or spraying. Plants, plants, at least one selected from the group consisting of the strain according to any one of claims 1 to 5, a culture medium of the strain, a microbial preparation containing the strain, and a microbial preparation containing the culture medium of the strain. A method for controlling phytopathogenic fungi comprising treating the surrounding area, or both. The method of claim 13, wherein the plant pathogenic fungus is Botryosphaeria dothidea .