KR20150093027A - Bacillus amyloliquefaciens KR-PBS6 as a novel strain with antifungal and enzyme activity - Google Patents

Abstract

The present invention relates to a novel bacillus amyloliquefaciens KR-PBS6 strain having antibacterial activity against plant pathogens and a use thereof.

Description

A novel strain of Bacillus amyloliquefaciens KR-PBS {Bacillus amyloliquefaciens KR-PBS6 as a novel strain with antifungal and enzyme activity}

The present invention relates to a novel strain of Bacillus amyloliquefaciens KR-PBS6 which has antibacterial activity against plant pathogenic bacteria and its use.

In order to solve the problem of food security along with the global population increase, the cultivation environment of the crop has also developed remarkably. However, crops such as wild animals, pests, and plant pathogens affect production, resulting in economic losses of up to 220 trillion won worldwide. In Korea, the three sides are surrounded by the sea. 65% of the country is a mountainous region and the land is narrow compared with the population. Recently, the cultivation area of horticultural crops has been gradually increasing by using the cultivation of facilities. , Cucumber, eggplant, pepper, strawberry, melon, tomato, melon, lettuce and leek. Organic synthetic pesticides and biological agents (biopesticides or microbial pesticides) are used as remedies for pests. Of these, organic synthetic pesticides were developed in the early 1900s and can be used to increase crop production, stabilize supply, and reduce labor force in a short period of time. However, due to the adverse effects such as soil pollution, groundwater pollution, accumulation of pesticides on agricultural products, and the emergence of resistant pathogens, the social demands of safety oriented, health oriented, and natural oriented are increasing in advanced countries in recent years, .

Therefore, since the Rio Declaration of the United Nations Conference on Environment and Development, in order to protect the global environment, abuse of chemical pesticides has been restricted and replaced with microorganisms themselves or biologically active pesticides using their functions. Biological pesticides are pesticides that are used to directly control microbes themselves or to control pests by using formulations containing them. These types of pesticides are classified into microbial insecticides, microbial fungicides and microbial herbicides. There are few toxicities, few effects on ecosystems, sustainable effects, and low investment costs. However, the efficacy of the drug is delayed, and the stability of the product and the difficulty in the production method become problems.

The microorganism preparation of the pesticide which is developed at this time shows the biological control effect by the antagonistic microorganism and the soil composting ability according to various enzyme activities. The principle of these microbial preparations has the main mechanism of action such as antibiotic action by antibiotics produced by them, and compaction through separation of residues in soil by various degrading enzymes.

Representative microbes currently registered for use as microbial agents include Bacillus sp., Candida sp., Lactobacillus sp., Lactococcus sp., Rhodobacter sp. Rhodobacter sp., Pseudomonas sp., And the like. As research on useful microorganisms continues, more and more microorganisms are registered as usable microorganisms every year. It is predicted that the annual growth rate will be maintained at 1520%, which is about 2 trillion won market until the early 2000s. Most of the strains that can be developed as a product are Fusarium solani ( Botrytis spp.), Botrytis cinerea (gray mold), Rhizoctonia solani ( constellation ), Stemphylium herbarum (rot, leaf blight), Diaporthe ambigua (stem blight), Colletotrichum acutatum (anthrax), Phytophthora cactorum (plague), Phytophthora capsici (plague), Didymella bryoniae (vine blight disease) and the like, but it has not yet occupied a large market. As mentioned above, there is a tendency to use biopesticides for environmentally friendly agriculture. Among them, various microorganisms having antagonistic action have been studied in order to reduce damage to plant pathogens. However, various pathogenic microorganisms Microorganisms having excellent antagonistic action that can be effectively controlled have not been reported.

Under these circumstances, the present inventors have made intensive efforts to develop microorganisms having an excellent antagonistic action that can effectively control various pathogenic microorganisms causing diseases of various crops. As a result, various pathogens, especially anthrax, The present inventors have developed a strain showing excellent antagonistic activity and protease, cellulase and α-amylase enzyme activity in blight, stem blight, rot fungus, gray mold disease and root rot disease, and completed the present invention Respectively.

One object of the present invention is to provide a process for the preparation of Fusarium solani solani , Botrytis < RTI ID = 0.0 > cinerea , Rhizoctonia solani , Stemphylium herbarum), Deirdre O Bhikkhus podeo cancer (Diaporthe ambigua , Colletotrichum acutatum) and Didier melanoma in Brioni (for one or more pathogens selected from the group consisting of Didymella brtoniae) which is characterized by having antimicrobial activity, the accession No. KCTC18239P Bacillus amyl quinone Lowry Pacific Enschede (Bacillus amyloliquefaciens ).

Another object of the present invention is to provide a microorganism preparation for controlling plant diseases, comprising the strain, the spore of the strain, the culture of the strain, or a mixture thereof as an active ingredient.

It is another object of the present invention to provide a method for controlling plant diseases using the microbial agent.

In some embodiments for achieving the above object, the present invention is Fusarium solani (Fusarium solani , Botrytis < RTI ID = 0.0 > cinerea , Rhizoctonia solani , Stemphylium herbarum), Deirdre O Bhikkhus podeo cancer (Diaporthe ambigua , Colletotrichum acutatum) and Didier melanoma in Brioni (for one or more pathogens selected from the group consisting of Didymella brtoniae) which is characterized by having antimicrobial activity, the accession No. KCTC18239P Bacillus amyl quinone Lowry Pacific Enschede (Bacillus amyloliquefaciens ).

The term "bacillus" in the present invention refers to aerobic or flowable anaerobic gram-positive bacilli widely distributed in nature, and Bacillus amyloliquefaciens as a microorganism belonging to the genus Bacillus. The present inventors have identified a novel strain belonging to Bacillus amyloliquefaciens as a microorganism which can be used as a pesticide because of its excellent antimicrobial activity and enzyme activity.

Specifically, in the present invention, in order to identify the microorganisms which can be used as biological pesticides with excellent antimicrobial activity and enzymatic activity, soil having good compostability was collected from Jeongeupseong-si, Jeonbuk Province and diluted to 10 ⁵ with 0.85% NaCl, Respectively. After that, pure water was separated from the LB medium and stored in a -80 ° C freezer. Then, the cells were cultured for 2 days using LB medium, centrifuged at 10,000 rpm for 15 minutes, and the supernatant was filtered with a 0.25 μm filter to separate the culture medium and cells. Thereafter, a hole was drilled in the PDA medium, and 100 μl of the purified culture was dispensed. After that, 100 μl of the purified culture was dispensed, and then the cells were treated with Fusarium solani, Borotitis cinerea, Riyotonia solani, Stempylium herbarum, Diaphora ambuchea, Riccum aquatum, and Didimella brioni were cultured to isolate strains having excellent antimicrobial activity against all of the strains. Finally, a strain with excellent protease, cellulase, and alpha-amylase activity was finally selected using LB medium using skim milk powder, carboxymethyl cellulose, and Soloblastase. As a result of the identification of this strain, a new strain belonging to Bacillus amyloliquefaciens Respectively. Thus, the present inventor named this Bacillus amyloliquefaciens KR-PBS6 and deposited it on January 30, 2013 in the Gene Bank of Korea Research Institute of Bioscience and Research and received the accession number KCTC18239P.

In accordance with one aspect of the present invention, the Bacillus of the present invention, amyl quinone Lowry Pacific Enschede KR-PBS6 strain is Fusarium solani (Fusarium solani), Botrytis cine LEA (Botrytis cinerea), Rhizoctonia solani (Rhizoctonia solani , Stemphylium herbarum), Deirdre O Bhikkhus podeo cancer (Diaporthe ambigua , Colletotrichum acutatum and Didymella < RTI ID = 0.0 > brtoniae ) strains (Figs. 1 to 7; and Table 2).

The Fusarium solani is a causative organism of mosaicism and root rot, Botrytis cinerea is a causative organism of gray mold, Riyotonia solanii is a causative agent of drowning and root rot, Stempylium herbarum is a causative organism of rot, The Bacillus amyloliquefaciens KR-PBS6 of the present invention is a causative organism for blight, and Diaphora amycigusa is a causative organism of stem blight. Colletoricum acutatum is a causative organism for anthrax, It is useful as a biological control agent against anthracnose, dwarf, pollen, leaf blight, vine blight, stem blight, rot, gray fungus or root rot.

In addition, according to one embodiment of the present invention, the strain of the present invention produces a protease that is a protease, a cellulase that is a fibrinolytic enzyme, and an alpha-amylase that is a starcholytic enzyme, as well as a biosurfactant effect Table 3).

Therefore, the Bacillus amyloliquefaciens KR-PBS6 of the present invention has a biological control effect and a soil compaction effect through enzyme activity, and is a strain that can be usefully used as a pesticide for environmentally friendly agriculture.

Here, the biological pesticide is defined as a commercial product of microorganisms isolated and collected in a natural environment in order to control plant pathogenic bacteria of crops.

On the other hand, peach blight, root rot, gray mold, melancholy, rot, leaf blight, stem blight, anthracnose and vine blight are the major diseases causing crop damage.

Among them, anthracnose of the plant is caused by choletotrichum acutatum and destroys the leaves, stems, fruits, flowers, etc. of the crops. It is one of the most serious diseases in pepper crops, an indispensable seasoning vegetable in Korean food Han KS et al., Res. Plant Dis. (2009) 15 (2), 88-93).

Root rot disease is a symptom in which part or all of the roots of a plant are decomposed or rotted, and its pathogen Rhizoctonia solani and Fusarium solani are damaged not only because the internal structure of the root is destroyed, but also because it is low in the ability of the root to absorb the root, so that the leaf is dropped and the fertilization rate is remarkably lowered (Kim WG, Korean J Plant Pathol. (1996) 12 (1), 21-32, Jeon HM, Isolation and Identification of Sickle Root Rot of Plants (2012) Master's Thesis, Sunchon National University).

Gray mold fungus is caused by Botrytis cinerea . It is a common disease occurring in the cultivation of plants. Generally, fruits and vegetables among crops are often caused by flower part after flowering, resulting in a great loss of fruit (Cheong SS et al., Kor J. Mycol. (2013) 41 (1) 38-41).

Aphrodisiac is a soilborne disease that can be caused by Rhizoctonia solani , in which part of the plant, for example the stem part, is constricted.

It is known to be a disease that can occur in almost all crops, especially in young seedlings that have germinated and have fallen to the root of the trunk of the edible grave. Fusarium solani ( Fusarium solani ) is known as a causative organism.

Stem blight disease is a disease that occurs in the main stem or branch of a tree and gradually becomes irregular with a dark brown lesion on the surface at first. The diseased part is depressed or swollen and the branch is dead. It is called. Staphylococcus is a diaphora ambigua ) are known to cause parasitism.

Vine blight disease is a plant disease caused by acanthosis, and occurs in leaves, stems, fruits, or fruits of watermelons, melons, melons, cucumbers, The leaves initially have small brown spots, and when they develop, they are enlarged to a large brown lesion (1 to 2 cm) of irregular shape. Small white spots appear on the stems and stomachs. When the stems grow, the lesions are enlarged along the stems. Severe stems all over and lesions form small black spots. As a representative pathogenic bacterium of the vine blight disease , Didymella bryoniae is known.

A rot disease is an irregular rotting discoloration of a diseased part, which can occur as parasitism of Stemphylium herbarum .

Leaf blight is a botanical disease that manifests as a wilting symptom of the leaf, and may appear particularly in onion, garlic, and onion by Stemphylium herbarum .

As described above, since the strain of the present invention exhibits a strong antagonistic action against pathogenic bacteria causing various plant diseases including anthracnose and root rot disease, when the strain is used as an environmentally friendly non-pesticide biological pesticide, It can be useful for eliminating toxic problems and can reduce the financial burden on each farm. In addition, due to the nature of the microorganism, it can be sustained and its long-term benefits can be seen in the future.

In another aspect, the present invention provides a microbial agent for controlling plant diseases, which comprises as an active ingredient the above-mentioned Bacillus amyloliquefaciens KR-PBS strain, spore of said strain, culture of said strain, or a mixture thereof do.

The above-mentioned Bacillus amyloliquefaciens KR-PBS6 is as described above, and the strain is selected from the group consisting of Fusarium solani, Borontis cinerea, Riyotonia solani, Stempylium herbarum, Diaphora ambuchea, It exhibits an excellent antimicrobial activity against all of the strains of E. coli, E. coli toricum acutum and D. melanburio, and thus can be used for the control of plant diseases caused by the bacteria. In addition, the microbial agent may be used as an overall microbial agent such as soil improving agent, but is not limited thereto, in addition to being used as a plant disease controlling agent.

In particular, the Bacillus amyloliquefaciens KR-PBS 6 according to the present invention can reduce the damage caused by contamination of plant pathogens that cause problems in cultivation of crops and has various enzyme activities, .

The damage caused by the contamination of the plant pathogenic bacterium is caused by the growth of Fusarium solani, Botrytis cinerea, Raiotonia solanii, Stemphilium herbarum, Diaphora ambuchea, Colletoricum acutum and Didimelabruronii .

In addition, as long as the antibiotic activity against the microorganism is exhibited, not only the microorganism but also the spore of the microorganism, the culture of the microorganism, or a mixture thereof may be included in the microorganism preparation.

In the present invention, the term "spore" refers to a reproductive cell such as a bacterium. For the purpose of the present invention, the spore refers to a reproductive cell of Bacillus amyloliquefaciens KR-PBS6. In the present invention, the spores of the Bacillus amyloliquefaciens KR-PBS 6 are selected from the group consisting of Fusarium solani, Borontis cinerea, Rai tononia solani, Stamphilium herbarum, Diaphora ambuchea, And / or the antimicrobial activity against Didimelebroronii, and can be used in a microbial preparation for controlling plant diseases.

In the present invention, the term "culture product" means a product obtained by culturing the strain, and includes cultured strains or both cultured strains.

On the other hand, the strain of the present invention can be used as a probiotic agent, preferably as a liquid probiotic agent or a live bacterial powder.

The Bacillus amyloliquefaciens KR-PBS 6 strain of the present invention may further contain an excipient and a diluent. The Bacillus amyloliquefaciens KR-PBS 6 strain may be used alone or in combination with other extracts, Can be used with byproducts.

The strain of Bacillus amyloliquefaciens KR-PBS 6 according to the present invention can be formulated in the form of a culture solution or powder according to a conventional method. Examples of excipients and diluents that can be included in the microorganism preparation using the strain include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Magnesium stearate, stevia extract, mineral oil and natural material extracts, and the like can be given as examples of water-soluble polymers such as calcium silicate, cellulose, microcrystalline cellulose, methylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, have.

Such a microbial preparation of the present invention is preferably, but not necessarily, diluted 500 to 1,000 times in water to be used for side spraying and root spraying.

In another aspect, the present invention provides a method for controlling a plant disease using the microbial agent. Specifically, the present invention provides a method of controlling a plant, comprising introducing the microbial agent into a plant or a growth environment of the plant.

The above microbial preparation and control are as described above.

Preferably, the botanical bottles may be selected from the group consisting of peach root rot, root rot, gray mold, rotten rot, rot fungus, leaf blight, stem blight, anthrax or vine blight.

It is preferable that the above-mentioned microorganism preparation is sprayed by using the foliar treatment method and the root of the plant roots at the ground surface of the plant at the time of clearing the soil before the preparation of the crop, 15 days after the planting, or just before the rainy season and the onset of the rainy season.

The Bacillus amyloliquefaciens KR-PBS 6 strain of the present invention has excellent antimicrobial activity and growth on plant pathogens causing anthracnose, constipation, pollinosis, leaf blight, vine blight, stem blight, rot disease, gray mold disease or root rot disease Inhibitory effect, and the enzymatic activity of protease, cellulase, and alpha-amylase is excellent, affecting soil composting ability. Therefore, the strain of the present invention is very useful as an environmentally friendly microbiological agent which can be used as a probiotic agent and a live bacterial powder to prevent destruction of an ecosystem.

Figure 1 shows the antimicrobial activity of the Bacillus amyloliquefaciens KR-PBS6 of the present invention against Fusarium solani .
Figure 2 shows the antimicrobial activity of the Bacillus amyloliquefaciens KR-PBS6 of the present invention against Rhizoctonia solani .
Figure 3 shows the antimicrobial activity of Bacillus amyloliquefaciens KR-PBS6 of the present invention against Botritis cinerea .
Figure 4 shows the antimicrobial activity of the Bacillus amyloliquefaciens KR-PBS6 of the present invention against Stemphylium herbarum .
Figure 5 shows the antimicrobial activity of the Bacillus amyloliquefaciens KR-PBS6 of the present invention against Diaporthe ambigua .
Figure 6 shows the antimicrobial activity of the Bacillus amyloliquefaciens KR-PBS6 of the present invention against Colletotrichum acutatum .
Figure 7 shows the antimicrobial activity of Bacillus amyloliquefaciens KR-PBS6 of the present invention against Didymella bryoniae .
FIG. 8 shows the enzymatic activity of the Bacillus amyloliquefaciens KR-PBS 6 of the present invention on protease, cellulase, and alpha-amylase.

Hereinafter, the present invention will be described in more detail with reference to the following examples. It should be noted, however, that the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example  1: Isolation of strains used

In this Example, as a test strain, Fusarium solani, Borontis cinerea, Raiotonia solani, Stamphilium herbarium, Diaphora ambigua, Colletoricum aquatum and Didymelbroronii were used as test strains and their antibacterial activity Soil samples were collected from Busan, Jeongeup, Jeonbuk Province in order to isolate the microorganisms. Samples were collected at depths of about 15 centimeters from the topsoil, and weeds and dead leaves were removed. This was sealed in a zipper bag and then transported to the laboratory at 0 to 4 ° C. 1 g of the collected sample was diluted to 10 ⁵ times with the diluted agar plate method, and 0.1 mL of the diluted solution was dispensed into the LB medium. The colonies were cultured in a 37 ° C. incubator for 2 days, Transferred to a medium (Table 1) and stored at -80 ° C.

The test strains were Fusarium solani, Bortithis cinerea, Raietonia solanii, Stempylium herbarium, Diaphora ambugua, Colletoricum aquatum and Didymelbroroni, and common to 7 strains Were selected as the best strains showing strong antibacterial activity. The strains of the selected strains formed circular colonies in the LB solid medium and appeared brown.

Medium component Concentration (g / L) Yeast extract 5.0 Tryptone 10.0 NaCl 10.0 Agar 15.0

Example  2: Identification of isolated strains

The nucleotide sequence of 16S ribosomal RNA was analyzed to identify the phylogenetic location of the selected strains. The selected strains were PCR amplified using modified colony PCR method. The primers used for 16S rRNA amplification were 27F (5'-AGAGTTTGATCCTGGCTCAG-3 ', SEQ ID NO: 1), 1492R (5'-GGTTACCTTGTTACGACTT-3', SEQ ID NO: 2), UP-1S (5'-GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA- (SEQ ID NO: 3), UP-2R (5'-AGCAGGGTACGGATGTGCGAGCCRTCNACRTCNGCRTCNGTCAT-3 ', SEQ ID NO: 4), the amplified product was purified using a PCR product purification kit (Qiagen) ) Was used to analyze the base sequence. As a result of analysis of the homology between the 16S rDNA of the final selected strain and the databases of DDBJ / NCBI / Genebank and Ribosomal database project, it was found that the strains belonging to the phylogenetic group including Bacillus sp. Were Bacillus amyloliquefaciens NBRC 15535T (AB255669), and more than 99% of the strains were found to be related.

On the other hand, the gyrB gene is expressed in Bacillus subtilis subtilis ) group, Bacillus subtilis subsp. Subtilis (B. subtilis subsp. subtilis), Bacillus piece you miss Fort (B. licheniformis), Bacillus amyl Lori rake Pacifico Enschede (B. amyloliquefaciens), the Bacillus art hand mouse (B. atrophaeus), Bacillus Belle Zen Systems (B. velezensis), Bacillus balises B. vallismortis , Bacillus subtilis subsp. B. subtilis subsp. The molecular genetic analysis was performed on the basis of the nucleotide sequence of the gyrB gene of the isolated strain, since it is the most useful information in order to grasp the intermediate relation between spizizenii and B. sonorensis . As a result, it was confirmed that it showed a flexible relationship of 99.07% with Bacillus amyloliquefaciens (DQ309294).

In addition, C _{15 : 0} anteiso (40.43%) and C _{15 : 0} iso (24.73%) were the major fatty acids in the analysis of the fatty acid composition of the fungus. Saturated fatty acids such as C _{14 : 0} and C _{16 : 14: 0 iso, C 16:} 0 iso, C 17: 0 iso , and C _17: showed the features containing a branched fatty acid, such as ₀ anteiso, Bacillus amyl Lowry quinolyl Pacifico main cell fatty acids Enschede type strain composition and FIG. It was confirmed to have a similar fungal fatty acid composition.

Thus, the strain of the present invention was finally confirmed to be a novel strain belonging to Bacillus amyloliquefaciens, which was named Bacillus amyloliquefaciens KR-PBS6. It was deposited with the Korea Collection for Type Culture (KCTC) on January 30, 2013 and granted KCTC18239P as a deposit number.

Example  3: Antimicrobial activity measurement against various plant pathogenic bacteria

Plant pathogenic fungi include Rhizoctonia solani AG-2-1 KACC 40123 (gluttony), Botrytis cinerea KACC 40573 (gray mold), Fusarium solani KACC 40384 (rootstock, root rot), Diaporthe ambigua KACC 41219 (Stalks blight), Colletotrichum acutatum KACC 40689 (anthrax), Didymella brtoniae KACC 40669 (vine blight), Stemphylium 7 kinds of test bacteria such as herbarum (rot disease, leaf blight) were distributed from the Korean Agricultural Microbiology Resource Center and the range of antibacterial activity of Bacillus amyloliquefaciens KR-PBS6 was examined.

Specifically, each pathogen was cultured in potato agar medium (PDA, Difco) and used as a seed culture. The culture of the isolate grown in LB medium was centrifuged and filtered with a 0.25 μm filter. Then, a pre-treated culture broth was poured into a hard agar broth with a diameter of 5 mm and humidified, and each pathogenic bacterium was cultured in the medium, and cultured at 25 ° C for 3 to 7 days.

Herein, the effect of the antagonistic microorganism on various pathogens was examined with or without support, and the results are shown in Table 2 and Figs. 1 to 7.

As a result, as shown in Table 2 and Figs. 1 to 7, the strain of the present invention showed excellent antimicrobial activity against all of the tested seven strains, and particularly, Diaporthe ambigua KACC 41219 (Stalks blight), Colletotrichum acutatum KACC 40689 (anthrax), Didymella brtoniae KACC 40669 (vine blight), Stemphylium The antimicrobial activity of the herbarum (rot, leaf blight) was particularly excellent.

Example  4: Isolate  Enzyme activity measurement

The enzymatic activity of the isolated strain was measured by measuring the size of the transparent ring by adding 1% skim milk to the LB medium and measuring the protease activity. Cellulase activity was measured by adding 0.5% of CMC to the LB medium for 24 hours The cells were stained with 0.2% Congo-red for 15 min and discolored with 1 M NaCl to measure the size of the yellow circle. The α-amylase activity was determined by measuring the size of the yellow circle formed by dropping Rugol's solution for 24 hours in a medium containing 1% soluble starch. The results are shown in Table 3 and FIG.

As a result, the strain of the present invention showed excellent results in protease, cellulase, and amylase activity.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Korea Biotechnology Research Institute KCTC18239P 20130130

<110> Korea Research Institute of Bioscience and Biotechnology <120> Bacillus amyloliquefaciens KR-PBS6 as a novel strain with          antifungal and enzyme activity <130> PA131319KR <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 27F Primer <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> 1492R Primer <400> 2 ggttaccttg ttacgactt 19 <210> 3 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> UP-1S Primer <400> 3 gaagtcatca tgaccgttct gcaygcnggn ggnaarttyg a 41 <210> 4 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> UP-2R Primer <400> 4 agcagggtac ggatgtgcga gccrtcnacr tcngcrtcng tcat 44

Claims (8)

Fusarium solani), Botrytis cine LEA (Botrytis cinerea), Rhizoctonia solani (Rhizoctonia solani , Stemphylium herbarum), Deirdre O Bhikkhus podeo cancer (Diaporthe ambigua , Colletotrichum acutatum and Didymella &lt; RTI ID = 0.0 &gt; brtoniae) of, accession No. KCTC18239P, it characterized in that it has an antimicrobial activity against one or more pathogens selected from the group consisting of Bacillus amyl quinone Lowry Pacific Enschede (Bacillus amyloliquefaciens) strain.
The strain according to claim 1, wherein the strain has an anticancer effect against anthrax, melanocytic, melanogaster, leaf blight, vine blight, stem blight, rot, gray fungus or root rot.
The strain of claim 1, wherein the strain exhibits protease, cellulase, and amylase activity.
A microorganism preparation for controlling plant diseases, comprising the strain of any one of claims 1 to 3, the spore of the strain, the culture of the strain, or a mixture thereof as an active ingredient.
The method of claim 4, wherein the microbial agent is Fusarium solani (Fusarium solani , Botrytis &lt; RTI ID = 0.0 &gt; cinerea , Rhizoctonia solani , Stemphylium herbarum), Deirdre O Bhikkhus podeo cancer (Diaporthe ambigua), Colle Thor rikum Aku tatum (Colletotrichum acutatum and Didymella &lt; RTI ID = 0.0 &gt; wherein the microbial agent has a controlling effect on a plant disease caused by at least one pathogenic bacterium selected from the group consisting of bacterium,
The microbial formulation according to claim 5, wherein the plant disease is at least one selected from the group consisting of anthrax, melancholia, pollen, leaf blight, vine blight, stem blight, rot, gray mold and root rot.
The microbial formulation according to claim 4, wherein the microbial agent exhibits a soil compaction effect.
A method of controlling a plant, comprising introducing the microbial agent of claim 4 into the plant or the growth environment of the plant.

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