KR20140064174A - New microorganism paraconiothyrium minitans s134, and microbial agent and biopesticide containing the same - Google Patents

Abstract

The present invention relates to a novel microorganism, Paraconiothyrium minitans S134 and a microbial formulation and microbial pesticide containing same. The novel microorganism, Paraconiothyrium minitans S134 (KACC 93156P) strain of the present invention is represented by sequence number 1 and controls the growth of pathogenic bacteria causing sclerotinia rot or white rot. According to the present invention, provided are a novel microorganism, Paraconiothyrium minitans S134 having an excellent control effect against pathogenic bacteria causing sclerotinia rot or white rot, and a microbial formulation and microbial pesticide containing the same, so that the occurrence of sclerotinia rot or white rot is inhibited and the number of plants can be increased.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel microorganism, paraconium sialium minintans S134, a microorganism preparation containing the microorganism, and a microbial pesticide containing the microorganism,

The present invention relates to a novel microorganism paracoimidium minitans S134, a microorganism preparation containing the microorganism, and a microbial pesticide containing the microorganism, and more particularly, to a method for isolating and isolating a strain inhibiting the growth of sclerotinia or black rot fungi, As an active ingredient, and a microbial pesticide.

Today, agriculture has been devoted to increasing food production through methods such as breeding, improvement of soil fertility, pest control, and weed removal, but persistent pesticide use and abuse are causing serious pollution of soil and river.

In addition, since these pesticides remain in crops, they cause problems such as toxicity, environmental pollution, and toxicity to humans and livestock. Therefore, the use of pesticides is now being strengthened in the world.

As concerns about these pesticides and the quality of life have increased, recently, biological control using microorganisms and plant extracts has been of interest as a new alternative to controlling pests of crops.

In particular, as a biosurfactant using microorganisms, antibiotics or toxins produced from soil microorganisms can be used to protect plants from infection by pathogens by utilizing antagonistic action of microbial pesticides or microorganisms themselves, which control pathogens or insects, And microbial fertilizers that promote microbial growth.

Such biological control can be used in place of chemical pesticides which are harmful to the environment, so that it is possible to provide safe agricultural products to consumers.

Sclerotinia < RTI ID = 0.0 > sclerotiorum ) is a pathogen that infects more than 400 plants and occurs worldwide. Especially, it is the major disease that causes loss of about 15% of yields in the cultivation of lettuce in winter, and sclerotia exists in soil for many years and acts as infectious source.

Therefore, there is a desperate need to develop a strain capable of efficiently acting on a pathogenic bacterium that causes scleroderma or black deciduous sclerotinia, and a method for mass-culturing it, without harming the plant.

As related prior arts, Korean Patent Publication No. 10-2009-0011139 (published on February 02, 2009, entitled "Novel Connie Limiminsansum 2 and Method for Controlling Plant Pathogens Using the same) 10-1148789 (registered on May 16, 2012, name: Microorganism for controlling scleroderma and its use).

DISCLOSURE OF THE INVENTION An object of the present invention is to isolate, isolate and isolate S. pneumoniae S134 strain, which is a novel strain showing antimicrobial activity against pathogenic bacteria of scleroderma or black decaying scleroderma, and to provide microorganism preparations and microbial pesticides containing the strain .

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

In order to achieve the above object, the novel microorganism paraconiosyrium minintans of the present invention minitans ) S134 (KACC 93156P) is represented by SEQ ID NO: 1 and is characterized by inhibiting the growth of pathogens causing scleroderma or black decaying scleroderma.

The new microorganism paraconothyrium minitans S134 (KACC 93156P) is isolated from soil and is used as an active ingredient of any one of microbial pesticide, seed coating agent, soil improving agent, composting homework, leaf fountain spraying agent or circulating spraying agent.

The gyunhaekbyeong occurs pathogens disk Loti California disk as thio volume (Sclerotinia sclerotiorum), disk Loti California US Norte, and any one or more of (Sclerotinia minor), black rot caused gyunhaekbyeong pathogen disk Loti volume sepia Bohrium (Sclerotium cepivorum ) is characterized.

In addition, the microorganism preparation of the present invention is characterized in that the new microorganism paraconothyrium minitans S134 (KACC 93156P) or a culture thereof as an active ingredient.

In addition, the microbial pesticide of the present invention can be used as a new microorganism paraconothyrium minitans S134 (KACC 93156P) or a culture thereof as an active ingredient, characterized in that the microbial pesticide is sprayed on the soil or inhabited in the soil to prevent scleroderma or black rot mycosis.

The present invention provides a novel microorganism paracoordium minitans S134 having an excellent control effect against scleroderma or black rot mycorrhage, a microorganism preparation containing the microorganism and microorganism pesticide containing the same, thereby suppressing the occurrence of scleroderma or black rot mycosis, It is possible to increase the quantity of the water.

Brief Description of the Drawings Fig. 1 is a 16S rDNA base sequence of the novel microorganism paracoordium minimalitus S134 strain of the present invention.
FIG. 2 is a phylogenetic map showing the 16S rDNA sequence of the novel microorganism Paracoonius minimalis S134 strain of the present invention.

The terms, techniques, and the like described in this specification are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified.

The new microorganism Paraconothyrium minitans strain S134 of the present invention is represented by SEQ ID NO: 1 and exhibits antimicrobial activity against pathogenic bacteria causing scleroderma or black decaying scleroderma , isolated from soil, .

That is, in order to select a new strain exhibiting a high antimicrobial activity against pathogenic microorganisms of scleroderma or black decaying scleroderma, the microorganisms isolated from the soil are inoculated into a PDA medium containing aureomycine, The isolates were selected for their ability to inhibit growth of pathogens causing scleroderma or black decay.

Analysis of the 16S rDNA obtained from the selected strain revealed that it was a novel strain of Paraconiothyrium minitans which was transformed into Paraconiothyrium minitans ) strain S134, and deposited this strain on the Agricultural Genetic Support Center of the National Academy of Agricultural Science and received the accession number KACC93156P on September 04, 2012.

The morphological characteristics of the new microorganism Paraconothyrium minitans S134 of the present invention are as follows: the morphological characteristics of the new microorganism Paraconothyrium minitans S134 are spherical or aspheric, with a diameter of 200 to 600 탆, and the pupil has a spherical or elliptical shape of 3.5 to 7.5 × 3.0 to 5.0 μm.

The above Paraconothyrium minitans ) The S134 strain consists of the entire nucleotide sequence shown in SEQ ID NO: 1 or a part thereof and is composed of an open reading frame (ORF) of 596 bp.

The new microorganism Paraconiothyrium minitans strain S134 of the present invention can be industrially used because it can be mass-produced in a short time when it is cultured under the following culture conditions.

The new microorganism grows well on the PDA medium as a culture medium. In particular, the culture medium preferably contains soluble starch as a carbon source, yeast extract powder as a nitrogen source, and salt as an inorganic salt. Preferably, the soluble starch is contained in an amount of 1 to 5% by weight based on the weight of the culture solution, the yeast extract powder is contained in an amount of 3 to 7% by weight of the culture solution, and the salt is contained in an amount of 0.001 to 0.1% The optimal conditions for the growth of the strain include 2% by weight of the soluble starch, 5% by weight of the yeast extract powder, 5% by weight of the culture solution, The salt is contained in an amount of 0.01% by weight based on the weight of the culture solution.

At this time, when the soluble starch is contained in an amount of less than 1% by weight, energy-generating elements are too small to slow the growth rate of the strain. If the soluble starch is contained in an amount exceeding 5% by weight, Thereby slowing the growth rate of the strain.

When the yeast extract powder is contained in an amount of less than 3% by weight, the growth and reproduction of the strain are slowed, and the specificity of the substrate to be selectively catalyzed is lowered. When the yeast extract powder is more than 7% by weight, excessive catalyst is added, There is a possibility that proper mixing is not achieved and at the same time strain of the strain is caused.

When the salt is contained in an amount of less than 0.001% by weight, the effect of the central role in metabolism is deteriorated, so that the growth rate of the strain is lowered. When the salt is contained in an amount exceeding 0.1% by weight, But rather slows the growth rate of the strain.

Also, under the culturing conditions, the strain can grow in the range of 5 to 32 ° C, and is best cultured at 15 ° C in particular.

The pH of the new strain is in the range of 4.0 to 9.0 and the optimum pH is 7.5. In the treatment for each kind of light, the mycelium of the new strain is cultured in the order of cancer treatment, fluorescence near-ultraviolet (NUV) treatment.

The microorganism of the new microorganism paracoimus sialium minantus S134 of the present invention or the culture thereof is used as an effective ingredient of a microorganism preparation for inhibiting scleroderma or black decaying scleroderma or microbial pesticide.

Such microbial agents and microbial pesticides are excellent in antimicrobial microbial pesticides, seed coating agents, microbial nutrients, soil improving agents, composting homework agents, foliar spraying agents, or antimicrobial agents since they are excellent in the survival period, survival germination rate, And may be used as a conventional spraying agent or the like.

The present invention will be described in more detail with reference to the following examples and experimental examples, but the scope of protection is not limited to the following examples and experimental examples.

<Example 1> Separation of novel antagonistic microorganisms of the present invention

Soil was collected from the lettuce rhizosphere of lettuce cultivation field in Yangpyeong-eup, Yangpyung-eup, Gyeonggi-do, Yangpyeong-eup, Gyeonggi-do, and the soil was allowed to settle by soil dilution method. The aureomycine (20mg / And cultured in a PDA medium.

Example 2 Identification of the novel antagonistic microorganism of the present invention

(1) Identification according to morphological characteristics

The size and shape of the isolates were examined and identified.

The results are shown in Table 1 below.

Investigation contents The novel strain of the present invention Punithalingam (1982) Barracks Dark brown, spherical or aspheric
Diameter 200 ~ 600㎛ Dark brown, spherical or aspheric
Diameter 150 ~ 600㎛ A planter Black brown, spherical or oval
3.5 to 7.5 × 3.0 to 5.0 μm Dark brown, spherical or oval,
4.0 to 7.0 占 2.5 to 3.5 占 퐉

(2) Molecular biology identification

In order to investigate the molecular biological properties of the bacterium isolated in Example 1, taxonomic similar species to the strain of the present invention were used, and the rDNA was compared and analyzed by ITS base sequence analysis.

In order to support the strain of the present invention as a novel strain, sequence analysis of the rDNA region of ITS (internal transcribed spacer) was carried out, and the results were as shown in Fig.

For DNA sequence analysis of the ITS region of rDNA, the DNA of the strain of the present invention was extracted and amplified using ITS1 and ITS4 primers. The amplification product was ligated into a vector, transformed into E. coli, and then cultured to extract plasmid DNA from the transformant, and the base sequence was analyzed.

The nucleotide sequences of the inventive strains were first aligned with the nucleotide sequences of other similar strains registered with the Gene Bank using CLUSTAL W software. In order to analyze the relationships between the strains, the ordered data sets were created using the MEGA 3.1 program. The bootstrap value of the branch was investigated through 1,000 resampling.

As a result of the phylogenetic analysis, it was proved that the strain was different from the other similar species as shown in Fig.

As described in the attached deposit certificate, the new microorganism named Paraconiothyrium m initans S134, which is a new soil microorganism, was deposited with the National Agricultural Research Service, And received accession number KACC93156P.

Example 3: Culture of a novel antagonistic microorganism of the present invention

In order to establish the culture conditions of Paraconiosyrium m initans strain S134, the mycelial growth of P. parasiticum Minantans S134 was measured by temperature, pH, light, and medium.

(1) Growth of S. parvum officinalis Min. Th134 strain by temperature

In order to investigate the temperature range of mycelial growth, the culture broth cultured on a potato dextrose agar medium (PDA) was detached with a 5 mm diameter cork roller and transferred to a 88 mm diameter Petri dish with PDA medium, The culture broth was cultured for 10 days in a thermostat controlled at 10, 15, 20, 22, 25, 28, 30, and 32 ℃ for three times.

Temperature
(° C) 5 10 15 20 22 25 28 30 32 Mycelial growth
(mm) 15.2 46.0 72.8 64.8 70.5 23.4 6.8 5.1 5.0

As shown in Table 2, it was confirmed that paracoimidium minitans S134 was able to grow at a temperature ranging from 5 to 32 ° C and best grown at 15 ° C.

(2) Growth of Paraconiolis minitans strain S134 by pH

In order to investigate the pH range at which the mycorrhizal growth of the Paracoidial Minimalans S134 strain was possible, the PDA medium adjusted from pH 4.0 to pH 9.0 at intervals of 9.0 was used to transfer the microorganism in the same manner as in the temperature experiment of the above (1) Then, the diameter of the microflora was measured after incubation in a 20 ° C thermostat for 10 days.

pH 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 Mycelial growth
(mm) 63.7 63.7 60.3 70.0 72.0 74.3 75.0 77.0 73.0 71.3 71.3

As shown in Table 3, it was found that p-conidic acid Minitans S134 was able to grow in the pH range of 4.0 to 9.0, and the optimum pH was 7.5.

(3) Growth of Paraconiolium minitans strain S134 by light type

PDA medium was divided into 88 mm diameter Petri dishes and irradiated with 20W fluorescent light for 12 hours and 20W near UV light (NUV ) Were irradiated at 12-hour intervals and cultured for 3 days at 20 ° C in a thermostat for 10 days to examine the length of mycelial growth

Treatment type Cancer treatment Fluorescent lamp Near-ultraviolet (NUV) Mycelial growth
(mm) 70.3 67.8 51.5

As shown in Table 4, paclitaxelium minitans S134 exhibited mycelial growth in the order of cancer treatment, fluorescent lamp and near ultraviolet (NUV), and it was found that mycelial growth was the best in cancer treatment conditions.

(4) Growth of S. parvum officinalis Minorans strain S134 by the medium

In order to determine the mycelial growth of each strain of Paraconianum minithans strain S134, each of the strains was divided into a 88 mm diameter Petri dish containing six kinds of mediums as shown in Table 5, The length of mycelial growth was examined after incubation for 10 days in a thermostat under dark conditions.

Badge type Medium composition Mycelial growth
(mm) PDA Difco PDA powder 39g, Dextrose 20g, Agar 15g 62.1 V8A
(v8juice agar) v8juice 200, CaCO ₃ 3g, agar 20g 51.8 Czapeck Dox agar ₂ g of NaNo ₃ , ₁ g of K ₂ HPO _4, 1 g of MgSO ₄ 0.5 g of 7H ₂ O, 0.5 g of KCl, 0.01 g of FeSO ₄ , 30 g of sucrose, 20 g of agar 36.1 MEA
(malt extract agar) malt extract 20g, Sucrose 20g Agar 20g 55.0 CMA
(corn meal agar) corn meal 17g, agar 20g 40.2 OMA
(oat meal agar) oat meal 60g, agar 20g 40.9

As shown in Table 5 above, it was found that parakonia minimalum S134 grew best in the PDA medium.

<Example 4> Sclerotinia or black decay Identification of scleroderma by parasitization on sclerotia of the causative bacteria

Para Connie Oh Solarium Mini Tansu to the disk Loti California Norilsk plant pathogens gyunhaekbyeong bacteria that can control the volume using the S134 strain thiosulfate (Sclerotinia sclerotiorum), Norilsk Zloty US California Norte (Sclerotinia minor ) and Sclerotium cepivorum , a black rot fungi, Sclerotium cepivorum , were investigated for the parasitism of S. pneumoniae S134 strain.

Sclerogenesis Pathogen + Sclerotinia sclerotiorum ,
Sclerotinia minor
Sclerotium cepivorum )

As shown in Table 6 above, it was confirmed that the Parconionium minitans strain S134 was parasitized on sclerotia of the above-mentioned plant pathogens and decontaminated sclerotia thereby to reduce the density of the infectious agent and to control scleroderma or black decline scleroderma.

<Example 5> Control effect of garlic black decay keratinocyte on packaging

Garlic (Namdo varieties) were sown in October 2011 to investigate the inhibitory effect of lettuce paracoordium minitans S134 on black rot mycotoxin in garlic cultivation of Taean.

Paraconium sialium minintens S134 was cultivated in oatmeal medium at 22 ° C for 15 days, and the spore suspension was diluted to 5 × 10 ^-6 ml ^-1 and treated once with 100 ml per week immediately after sowing and once in mid February.

The control agent was treated with wetting of Castella wax, and all treatments were repeated three times each.

For the determination of soil fixation and density change in the soil of garlic redistricting pArconium Minimalans S134 strain, CZA medium and Rose bengal agar (KH ₂ PO ₄ 1, MgSO ₄ 7H ₂ O 0.5, peptone 5, glucose 10, rose bengal 0.033, streptomycin sulfate 0.033, agar 20 g / L) and dilution plate culture method.

Processing contents Lee Byungju (%) Control (%) Growth rate (%) Paraconiolium minitans S134 (treated twice) 6.8 65.1 26.7 Castella waxing agent 0.4 97.9 46.9 No treatment 19.5 - -

As shown in Table 7, the control effect against Sclerotium cepivorum was confirmed. As a result, the new strain of Paracoignithium minintans strain S134 of the present invention showed an excellent control effect of 65.1% on the garlic harvester, And 26.7%, respectively.

Thus, S. parasiticum Minitans strain S134 is effective as an antidote to this black rot mycotic disease.

Processing contents The bacterial density (CFU × 10 ⁵ / g soil) 10.4 11.10 12.6 1.16 2.23 3.20 4.18 5.22 S134 2.6 1.2 1.7 29.4 17.5 3.5 4.7 1.5 Castella waxing agent 0 0 0 0 0 0 0 0 No treatment 0 0 0 0 0 0 0 0

As shown in Table 8, the excellent paracoordium minitans strain S134 of garlic black rot fungus control disease showed excellent fixing ability in the soil of the garlic redevelopment field, and the bacterial density was increased from the first treatment after the first treatment, .

Therefore, it is considered that the parakoniaricum minintans S134 of the present invention has a good fixing ability in the soil, so that it is possible to control the black decaying scleroderma only by treating only twice in the soil treatment.

<Example 6> Control effect of lettuce keratinocyte in a greenhouse

Lettuce was cultivated on a 50-tray tray port in the condition of 4.5 leaves, and cultured on oatmeal medium for 12 days. The cultured papillomavirus S. minantans strain S134 was treated with 5 ml per spore suspension concentration on days 7, 11, and 18 After 22 days, Sclerotinia sclerotiorum was inoculated into the soil of the lettuce with 2 barley cultivated at 20 ℃ for 8 days.

Spore suspension
Concentration (/ ml) After treatment with Paracoidus minimalum S134 strain
Rate of occurrence of scleroderma by age (%) 7 days 11th 18th 22nd 1 x 10 ⁶ 0.0 0.0 0.0 1.1 5 × 10 ⁶ 0.0 0.0 0 0 1 x 10 ⁷ 0.0 1.1 1.1 1.1 No treatment 4.4 28.9 52.2 55.5

As shown in Table 9, the spore suspension (5 × 10 ⁶ ml ^-1 ) of the Parconionis Minitans strain S134 exhibited excellent control effect so that no lettuce sclerotinosis was caused even 22 days after the treatment.

Example 7 Confirmation of Mycelial Germination and Decrease of Jangarin Formation in Sclerotia of Sclerotinia Virus of Parconioicum Minitans strain S134

In order to investigate the inhibition of bacterial growth of Paraconicosidium minitans strain S134 against sclerotinia, the culture was incubated for 12 days at 22 ° C in an oatmill medium of paconiconium minitans S134, and spore suspension (5 × 10 ^{6 -1} ), sclerotia of S. sclerotiorum was immersed for 20 minutes, and then 10 pieces of the PDA medium were placed in a 88 mm diameter Petri dish at regular intervals and cultured in a thermostat at 20 DEG C for 8 days , Germination of mycelial germs directly at sclerotium of sclerotinia bacteria and 15 days after treatment The parasitism of sclerotia of Paraconicosidium minitans strain S134 was examined.

In addition, the parakoniaricum minicans strain S134 was immersed in sclerotia ( S. sclerotiorum ) sclerotia for 20 minutes in a spore suspension (5 × 10 ⁶ ml ^-1 ) in the same manner as described above.

The treated sclerotia were pared by 10 per Erlenmeyer flask containing sand medium sterilized at regular intervals and examined for inhibition of the formation of the sacs after 60 days of irradiation with fluorescent lamps at 15 ° C for 12 hours / day.

Scleroderma Treatment concentration Mycelial germination rate (%) Peritoneal flap formation rate (%) Sclerotial infection rate (%) S. sclerotiorum 1 × 10 ⁶ ml ^-1 0 0 100 No treatment 100 100 0

As shown in Table 10, the S. pariquii S. minantus strain S134 infected all sclerotia of S. sclerotiorum and completely inhibited mycelial direct germination and sacral formation to inhibit the sclerotial activity of sclerotia .

Provided is a natural microorganism preparation and a microbial pesticide which can inhibit the occurrence of scleroderma or black decaying scleroderma by providing a new microorganism paraconiosporum minantus S134 having excellent control effect against causative bacteria of scleroderma or black decaying scleroderma as described above So that the number of crops can be increased.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. You can implement the examples. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

National Institute of Agricultural Science KACC93156P 20120904

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> NEW MICROORGANISM PARACONIOTHYRIUM MINITANS S134, AND MICROBIAL          AGENT AND BIOPESTICIDE CONTAINING THE SAME <130> P2012-0166 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 596 <212> DNA <213> Paraconiothyrium Minitans S134 <400> 1 tccgtaggtg aacctgcgga aggatcatta tccatcctta acaggtgcgg tcgcggcccc 60 tgggggctcc cccaggtggt aaggtgaaat accctatcgc gccgcatacc tgcatccttt 120 ttttacgagc acctttcgtt ctccttcggc ggggcaacct gccgctggaa cctgataaac 180 cttttttgca tctagtatta cctgttctga tacaaacaat cgttacaact ttcaacaatg 240 gatctcttgg ctctggcatc gatgaagaac gcagcgaaat gcgataagta gtgtgaattg 300 cagaattcag tgaatcatcg aatctttgaa cgcacattgc gccccttggt attccatggg 360 gcatgcctgt tcgagcgtca tctacaccct caagctctgc ttggtgttgg gcgtctgtcc 420 cgcctttgcg cgcggactcg ccccaaactc attggcagcg gtttttgcct cctctcgcgc 480 agcacaattg cgtctgcgag ggggcgtggc ccgcgtccac gaagcaacat taccgtcttt 540 gacctcggat caggtaggga tacccgctga acttaagcat atcaataagc ggagga 596

Claims (12)

1,
New microorganism Paraconothyrium m initans S134 (KACC 93156P) strain.
The method according to claim 1,
The new microorganism paraconiolum minithans strain S134 is isolated from soil,
New microorganism Paraconothyrium m initans S134 (KACC 93156P) strain.
The method according to claim 1,
The new microorganism paraconium sialium minantus strain S134 is a strain that inhibits the growth of pathogenic microorganisms causing scleroderma or black rot mycosis,
New microorganism Paraconothyrium m initans S134 (KACC 93156P) strain.
The method of claim 3,
The gyunhaekbyeong occurs pathogens thio volume (Sclerotinia sclerotiorum) to disk Loti California disk, disk Loti California US Norte (Sclerotinia minor ) &Lt; / RTI &gt;
New microorganism Paraconothyrium m initans S134 (KACC 93156P) strain.
The method of claim 3,
Wherein the black rot fungus pathogen is a sclerotium cepivorum ,
New microorganism Paraconiothyrium minitans S134 (KACC 93156P) strain.
The method according to claim 1,
The new microorganism Parconionis Minimalans S134 strain is used as an active ingredient of any one of microbial pesticides, seed coating agents, soil conditioners, composting homeworks, foliar spraying agents,
The new microorganism paraconothyrium minitans) S134 (KACC 93156P) strain.
A microorganism preparation for inhibiting the occurrence of scleroderma or black decaying scleroderma comprising the microorganism Paraconiothyrium m initans strain S134 of any one of claims 1 to 6 or a culture thereof as an active ingredient.
8. The method of claim 7,
The culture solution contains a soluble starch as a carbon source in a culture medium of PDA, a yeast extract powder as a nitrogen source, and a salt as an inorganic salt, and a microorganism preparation for inhibiting scleroderma or black decaying scleroderma.
9. The method of claim 8,
Wherein the soluble starch is 1 to 5% by weight based on the weight of the culture, the yeast extract powder is 3 to 7% by weight based on the weight of the culture, and the salt is 0.01 to 0.1%
Microorganism preparation for inhibiting scleroderma or black decaying scleroderma
10. The method of claim 9,
Wherein the culture liquid contains 2% by weight of the soluble starch, 5% by weight of the yeast extract powder, and 0.01% by weight of the salt, based on the weight of the culture liquid.
Microorganism preparation for inhibiting scleroderma or black decaying scleroderma.
A microorganism pesticide for inhibiting the occurrence of scleroderma or black decaying scleroderma comprising the microorganism Paraconiothyrium m initans strain S134 of any one of claims 1 to 6 or a culture thereof as an active ingredient.
A method for controlling scleroderma or black decaying sclerotia by spraying the microorganism pesticide of claim 11 or spreading it in soil.

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