KR101653959B1 - Pseudozyma churashimaensis RGJ1 strain isolated from pepper plant and uses thereof - Google Patents

Abstract

The present invention relates to a strain Pseudozyma churashimaensis RGJ1, which induces induced systemic resistance to a plant pathogen or a plant virus, which is isolated from a pepper plant, a strain of the above strain or a culture thereof as an active ingredient A method for producing a microorganism preparation comprising the step of culturing the strain and a method for producing the microorganism preparation by immersing the strain or a culture thereof in a seedling of a plant, A method for controlling a disease of a plant, comprising the step of foliar treatment.

Description

Pseudozyma churashimensis strain RGJ1 isolated from pepper plants and its use {Pseudozyma churashimaensis RGJ1 strain isolated from pepper plant and uses thereof}

The present invention relates to a Pseudomonas sulcatus RGJ1 strain isolated from a pepper plant and a use thereof, and more particularly, to a method for inducing an induced systemic resistance to a plant pathogen or a plant virus isolated from a pepper plant Pseudozyma < RTI ID = 0.0 > churashimaensis RGJ1), a method for producing a microorganism preparation comprising the above strain or a culture solution thereof as an active ingredient for controlling a plant disease or increasing the yield of a plant, a step of culturing the strain, and a method for producing the microorganism preparation, Which comprises immersing a seedling of a plant, treating the soil, or foliar treatment on the ground surface.

Currently, chemical synthesis pesticides are mainly used for inhibiting the generation and control of plant pathogenic bacteria. However, these synthetic pesticides are destructive to ecosystems, cause human toxicity due to residuals, and are very likely to cause various diseases such as carcinogenesis and anomalies, and their use is limited. Therefore, research on the development of environmentally friendly biological pesticides that can replace synthetic pesticides is being actively conducted. One of the methods that has been tried by many researchers to solve the problems of chemical control method is biological control method using microorganisms. The biological control method using microorganisms has recently been studied as an alternative to the chemical control method for the control of the diseases of the crops as the consumers' awareness of the food safety has increased and the preference for environmentally friendly agricultural products has increased rapidly.

An important disease of pepper is known as a botanical disease which threatens the production of pepper, such as an infectious disease of soil infectious disease, anthracnose, powdery mildew, and bacterial spotty disease which occur in the ground part. Pepper is one of the most frequently treated crops because it is not easy to control because the length of cultivation period is long and the pattern of disease development is very different according to the change of weather. Therefore, it is important to select microorganisms with excellent disease control effect and use them for disease control in order to produce environmentally friendly pepper. Most of the microorganisms developed to control pepper disease have been developed for the control of pepper blight. Shen et al. Reported that Serratia plymuthica was effective in controlling pepper blight (Shen et al., Plant Pathol. J., 21, 64-67, 2005). However, there are few reports of microorganisms that have excellent control effect against the topical diseases of pepper.

Korean Patent Laid-Open Publication No. 2009-0105726 discloses' Bacillus megaterium 22-5 strain for controlling bacterial spotty disease and anthracnose of pepper ', Korean Patent Publication No. 2012-0075936 discloses' Soil-derived Bacillus genus PB25 strain and Quot; Pseudosyma ( RGJ1 ) ", which induces induced systemic resistance to plant pathogens or plant viruses isolated from pepper plants as in the present invention, churashimaensis RGJ1) strain.

The present invention has been made in view of the above-mentioned needs, and an object of the present invention is to provide a Pseudosyma ( RGJ1) churashimaensis RGJ1) induces an induced systemic resistance to a bacterial pathogenic bacterium or plant virus of a pepper plant and has a plant disease control effect, thus completing the present invention.

In order to solve the above problems, the present invention relates to a method for inducing induced systemic resistance to plant bacterial pathogens or plant viruses isolated from pepper plants by using Pseudozyma ( RGJ1) churashimaensis RGJ1).

The present invention also provides a microorganism preparation for controlling a plant disease comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a microorganism preparation for increasing the yield of a plant comprising the strain or a culture thereof as an active ingredient.

In addition, the present invention provides a method for producing a microorganism preparation comprising culturing the strain.

In addition, the present invention provides a method for controlling a plant disease, comprising immersing the strain or a culture thereof in a seedling of a plant, treating the soil, or foliar treatment on the ground.

The strain Pseudozyma churashimaensis RGJ1 isolated from the pepper plant of the present invention induces an induced systemic resistance to a bacterial pathogen or a plant virus of a pepper plant to cause a plant disease control effect Respectively. The strain of the present invention can be used as a bio-pesticide that is environmentally friendly and free from the risk of human toxicity due to environmental pollution and pesticide residues caused by side effects of pesticides, and can be used for controlling pesticidal fungi of pepper plants and increasing plant disease resistance Therefore, it can be used industrially.

Figure 1 shows the localized sites sampled to separate live yeast from pepper and the yeast species isolated at that location.
Fig. 2 shows the results of primary analysis of the inducibility of yeast strains surviving in red pepper. RGJ1: Pseudozyma churashimaensis), RGJ5: Cryptosporidium Caucus Magnus (Cryptococcus magnus ), GS5: Pseudozyma aphidis ), GS6: Pseudozyma tsukubaensis )
FIG. 3 shows a secondary analysis result of induction resistance to RGJ1, a yeast strain isolated from red pepper. BTH (benzothiadiazole): inducible resistance control, Kanamycin: direct antiseptic control antibiotic
FIG. 4 shows the result of confirming the induction resistance of the red pepper treated with the yeast RGJ1 of the present invention to the bacterial spotty pathogen in the field.
Fig. 5 shows the result of confirming the virus-induced resistance of red pepper treated with the yeast RGJ1 of the present invention in the field.
FIG. 6 shows the result of confirming the virus-induced resistance of the red pepper treated with the yeast RGJ1 of the present invention in the field by quantifying the virus through qPCR. CMV: Cucumber Mosaic Virus, BBWV: Broad Bean Wilt Virus,
Fig. 7 shows the result of confirming the virus-induced resistance of red pepper treated with the yeast RGJ1 of the present invention in the field by quantification of virus through qPCR. PepMoV: Pepper mottle virus, PMMoV: Pepper mild mottle virus,
FIG. 8 shows the results of analysis of the expression levels of PR4 and PR5, genes involved in inducing resistance upon pathogen entry into red pepper treated with yeast RGJ1 of the present invention.
Figure 9 shows the increase in red pepper yield according to the treatment of the yeast RGJ1 of the present invention in the field.
Fig. 10 shows the results of confirming the leaf fixing ability of the yeast strain RGJ1 in the greenhouse.

In order to accomplish the object of the present invention, the present invention provides a method for inducing induced systemic resistance to a plant pathogen or a plant virus, which is isolated from a red pepper plant, wherein the Pseudozyma ( RGJ1) churashimaensis RGJ1). The above-mentioned Pseudomonas rhizomesis RGJ1 was deposited at the Korea Biotechnology Research Institute on Nov. 6, 2014 (Accession No .: KCTC18338P).

In a strain according to an embodiment of the present invention, the plant pathogenic bacteria or plant viruses may be pathogens or viruses for various plants, preferably, but not limited to, pepper plant pathogens or pepper plant viruses. The plant pathogenic bacterium may be a homogeneous bacterial pathogen, preferably homogeneous bacterial spotty bacterium, more preferably bacterial spotty pathogen ( Xanthomonas axonopodis pv. vesicatoria). < / RTI > The pepper plant viruses are selected from the group consisting of cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), bovine spongiform encephalopathy virus (BBWV), chrysanthemum virus virus (CMMV), sweet potato Y virus (PVY) Viruses such as Cucumber Mosaic Virus (CMV), Broad Bean Wilt Virus (BBWV), pepper sting virus (PepMoV), pepper (PepMoV) Light stain virus (PMMoV), and the like.

The present invention also provides a microorganism preparation for controlling a plant disease comprising the strain or a culture thereof as an active ingredient. The plant disease may be caused by a plant pathogen or a plant virus, preferably, but not limited to, caused by a pepper plant pathogen or a pepper plant virus. The pepper plant pathogen or the pepper plant virus is as described above.

The microbial agent may be Pseudozyma ( RGJ1 ), which induces induced systemic resistance to plant pathogens or plant viruses isolated from the pepper plants of the present invention, churashimaensis RGJ1) strain or a culture thereof may be contained as an active ingredient. The microbial formulation according to the present invention may be prepared in the form of a liquid fertilizer, and may be added in the form of a powdered powder or may be granulated by formulating it into a powdery fertilizer. However, the formulation is not particularly limited. In other words, it can be formulated as biological fertilizer to overcome this in eco - friendly organic agriculture with limited supply of chemical fertilizer.

The present invention also provides a microorganism preparation for increasing the yield of a plant comprising the strain or a culture thereof as an active ingredient. The plant may be, but is not limited to, a pepper plant. When the strain of the present invention was treated with pepper plants, the yield of the red pepper fruit was increased by about 30% as compared with the control.

In addition, the present invention provides a method for producing a microorganism preparation comprising culturing the strain. The yeast strain may be cultured by any method known in the art and is not particularly limited to a specific method.

In addition, the present invention provides a method for controlling a plant disease, comprising immersing the strain or a culture thereof in a seedling of a plant, treating the soil, or foliar treatment on the ground. The plant disease may be caused by a plant pathogen or a plant virus, preferably, but not limited to, caused by a pepper plant pathogen or a pepper plant virus. The pepper plant pathogen or the pepper plant virus is as described above.

The method for controlling the disease of the plant can be carried out by immersing the culture medium in which the microorganism is cultured and the microorganism preparation using the microorganism into the seedling of the plant, treating the soil, or treating the plant with foliage. In the case of immersion, the culture medium and the microbial agent may be poured into the soil around the plant or the seed may be immersed in the culture medium and the preparation.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1. Isolation of surviving yeast strains from red pepper

Leaves were collected from red pepper fields in Cholla province and Cholla province in order to isolate the yeasts that play a useful role in red pepper. From September to November, 2013, the leaves of the middle part of the red pepper were harvested in the areas of Geoje, Goseong, Hamyang, Jinan, Namhae and Suncheon. A 10-mm-diameter cork ball was cut into a circular disk, and three pepper blade discs were placed in a 1.5-ml tube containing sterilized water (1 ml), and zirconia beads (small beads with a diameter of 1 mm) So that they were better separated and vortexed in sufficient time. The leaf discs were diluted with 1/10, 1/100, and 100 μl each were plated on agar plate medium. In order to isolate only the yeast strains, the used agar medium was prepared by adding rifampicin (rifampin capsules; Yuhan), which is an antibiotic to prevent bacteria from growing, to a final concentration of 100 μg / ml in YPD medium Respectively. Yeast colonies from the diluted and plated plates were separately subcultured and grown into pure single cells and subjected to direct ITS sequencing (Genotech) analysis. The sequences of the ITS sequencing primers used are: ITS1 (forward): 5'-TCC GTA GGT GAA CCT TGC GG-3 '(SEQ ID NO: 1), ITS4 (reverse direction): 5'-TCC TCC GCT TAT TGA TAT GC-3 '(SEQ ID NO: 2)).

After that, the leaves were collected from the pepper field in Jeolla Province (Jean - Cheonan - Gosung - Tongyoung - Geoje - Namhae - Suncheon) region and identified through sequencing of isolated yeast. Various yeasts survived in the leaves of pepper. Among them, yeasts of Pseudozyma were slightly dominant, and a large amount of them survived (FIG. 1).

Example  2. Primary resistance analysis of inducible resistance to live yeast strains in pepper

In order to confirm the ability of the isolated yeasts to induce resistance to red pepper, a primary strain selection experiment was conducted in the greenhouse. Three days after planting, 7 days old red pepper (new PR, red pepper) was transferred to a 50 - port pot. After 3 weeks, except for cotyledon, the yeast strains were sprayed on the top part of the red pepper seedlings. At this time, the treated microorganisms were suspended in sterilized water for 3 days in a solid medium and sprayed with 50 ml per one hot pepper seedlings at OD600 = 1 (10 ⁶ CFU / ml). After 7 days from the treatment of the microorganism, the bacterial spotty pathogen ( Xanthomonas axonopodis pv. Vesicatoria) of pepper was inoculated into the back of the leaf of the pepper with a syringe by direct injection by setting OD600 = 0.01. Three pathogens were inoculated on each leaf of the pepper seedlings, and after 5 ~ 7 days of symptoms began to appear, the pathogen was examined and the resistance against pathogenic bacteria was confirmed. The disease was investigated by randomly dividing disease severity from 0 to 5 (0: no symptoms were observed, 1: leaves were slightly cloudy greenish than the original color) , 2: yellowish yellowing of the leaf, 3: black part of the yellow part, 4: more than 1/3 of the leaf turned black, 5: all of the leaf turned black, and fainting). Four leaves per pepper seedlings were investigated separately, and the number of symptoms was examined through a total of 5 repetitions.

7 days after the inoculation of the pathogens, the results of the investigation revealed that the yeast isolated from Geojedo Island, Pseudozyma ( RGJ1) churashimaensis RGJ1) yeast showed the best disease resistance effect. Compared with water treatment, 70% symptom decreased, which showed the same significance as the treatment of chemical BTH positive control (Fig. 2).

Example  3. Second-order analysis of the induction resistance of yeast strains isolated from red pepper

Among the yeast isolates used in the first-order induction-resistance assay, the Pseudozyma strain churashimaensis RGJ1) were investigated to determine the difference of induction resistance according to the treatment method. When the cultured broth of yeast RGJ1 was adjusted to OD600 = 1 and sprayed on the upper leaves of the pepper under the same conditions as the first induction resistance test, the difference in the culture conditions when the culture was directly applied to the soil was compared, BTH, a chemical that causes resistance, and water treatment in the greenhouse as a control. Further, in order to ensure that the direct bactericidal activity by the yeast RGJ1, after the Xav. Of pepper bacterial spot patterns pathogens on plate with a diameter of 90mm pre-plated according to OD600 = 0.5, placed topped that of above 10mm diameter paper disk, (Fig. 3), in which a sterilization effect was exhibited by culturing the cells at 30 DEG C by treating the same amount of yeast RGJ1 (20 microliters adjusted to OD600 = 1), BTH (1 mM), antibiotic kanamycin 25, .

Similar to the results of the primary resistance test, disease resistance was induced, and no difference was observed between spraying on the leaves and direct treatment on the soil. In addition, as a result of the confluent culture, it was confirmed that no transparent ring was formed around the yeast RGJ1, so that the bactericidal effect directly killing the pathogenic bacteria was not found, and it was confirmed to induce resistance (FIG. 3).

Example  4. In the field, RGJ1 Of red pepper treated Bacterial spotty disease Identification of Inducible Resistance to Bacteria

The ability of induction resistance in pepper field packaging was examined for the selected yeast strains in the laboratory and in the greenhouse. (36 ° 8 '50.81''N, 127 ° 29' 29.20 '' E) in Geumsan-gun, Chungnam Province. In June 2014, yeast RGJ1 broth was sprayed onto the ground surface of a month-old red pepper at a concentration of OD600 = 1 (10 ⁷ CFU / ml) under the same conditions as in the greenhouse experiment. As a positive control, BTH with inducible resistance was sprayed at a concentration of 1 mM and treated with water as a control. The length of the treatment line in the randomly divided treatment area was 8 meters, and the distance between the red pepper and the red pepper was 40 cm, which included 20 weeks of pepper per treatment. The culture medium and the control material per one treatment were treated in 2 liters, and the treatment was repeated four times at random. Ten days after the treatment, the bacterial spotty pathogen ( Xanthomonas axonopodis pv. Vesicatoria) was diluted to OD600 = 0.01 and inoculated with a syringe pierced on the back side of the leaf so as to enter the leaf of the red pepper to a half. Five pepper leaves were inoculated on each leaf, and 10 seedlings were inoculated per treatment. After 7 days, the symptoms were observed. The degree of the disease was divided into 0-5 by the same method as the confirmation experiment in the greenhouse in front of the disease investigation.

As a result of the field test, resistance was induced by the spray treatment of yeast in the same manner as in the indoor experiment, and the symptom of the bacterial spotty pathogen was reduced by 50% (FIG. 4).

Example  In the field, RGJ1 Induced Resistance of Virus-Treated Pepper to Virus

In the field experiment, 5 fresh leaves of red pepper that had passed 60 days after treatment with yeast were harvested in liquid nitrogen, rapidly frozen, ground into a mortar, and extracted with an RNA extraction kit (RNeasy kit ; QIAGEN). The extracted RNA was synthesized with cDNA and RT-qPCR, which is a method of quantifying the amount of virus with qPCR (CFX connect Real-Time System; BIO-RAD), was performed using the synthesized cDNA. The extent of infection was quantified using specific primers for naturally occurring cucumber mosaic virus (CMV) and broad bean virus (BBWV). The conditions of qPCR and the primers used are as follows. CMV coat protein forward: 5'-CGTTGCCGCTATCTCTGCTAT-3 '(SEQ ID NO: 3), CMV coat protein reverse direction: 5'-GGATGCTGCATACTGACAAACA-3' (SEQ ID NO: 4), BBWV forward: 5'- AATGAAGTGGTGCTCAACTACACA- 3 '(SEQ ID NO: 5), BBWV reverse direction: 5'-TTTTGGAGCATTCAACCATTTGGA-3' (SEQ ID NO: 6). In addition, the degree of infection was quantified using specific primers of peppermint virus (PepMoV, Pepper mottle virus) and pepper mild mottle virus (PMMoV, Pepper mild mottle virus). The conditions of qPCR and the primers used are as follows. (SEQ ID NO: 7), PepMoV reverse direction: 5'-CAAGCAAGGGTATGCATGT-3 '(SEQ ID NO: 8), PMMoV forward direction: 5'-ACAGTTTCCAGTGCCAATCA-3' (SEQ ID NO: 9), PMMoV Reverse direction: PepMoV forward: 5'-AAGATCAGACACATGGA- : 5'-AAGCGTCTCGGCAGTTG-3 '(SEQ ID NO: 10).

In the virus disease investigation, the fresh leaves of the pepper were compared with each other, and it was confirmed that the symptom was vigorous in the water treatment, whereas the symptom was weak in the yeast treatment (Fig. 5). In addition, both of the representative viruses such as CMV and BBWV showed a decrease in the amount of virus by about 10 times as compared with that of water treatment, as a result of the quantification of virus through qPCR (Fig. 6) . In addition, both PepMoV and PMMoV viruses were found to be infected with viruses by yeast treatment (Fig. 7).

Example  6. Yeast of the present invention RGJ1 Analysis of Expression Levels of Inducible Resistance-Related Gene in Pathogen-Entering Pepper Treated with

In order to investigate the mechanism of action of yeast, in order to investigate the amount of gene expression for the study of bacterial spotty pathogen in field experiment, ① before the inoculation of the pathogen ② at the time of inoculation of the pathogen ③ after the inoculation of the pathogen, , And the RNA was extracted by the same method as the virus quantitation method, and the expression amount of the resistance-related genes of the red pepper was examined through cDNA synthesis and qPCR. Expression of PR gene as a representative induction resistance gene of pepper was confirmed. The conditions of qPCR are the same as virus quantification and the primer information used is as follows. CaPR4 forward: 5'-AACTGGGATTTGAGAACTGCCAGC-3 '(SEQ ID NO: 11), reverse: 5'-ATCCAAGGTACATATAGAGCTTCC-3' (SEQ ID NO: 12), CaPR5 forward: 5'-CTCCACAAGAAACAAGGCA-3 '(SEQ ID NO: 13), reverse: 5 '-GTACGAAGCACGCACACAA-3' (SEQ ID NO: 14).

The expression levels of PR4 and PR5, which are resistance genes, increased significantly at 6 hours in the yeast-treated area, indicating that the priming effect was faster in response to pathogens invasion (Fig. 8).

Example  7. Yeast of the present invention RGJ1 By treatment Field pepper yield survey

After confirming the induction resistance in the field, the yield of pepper was examined in 20 individuals of each treatments and the number was confirmed. A total of four iterations were performed. It was confirmed that the yield of the yeast RGJ1 was increased by about 30% as compared with the control (Fig. 9).

Example  8. Yeast in the greenhouse RGJ1 Identification of leaf fixation ability

In order to confirm the ability of the yeast strain Pseudomonas sylvestris RGJ1 isolated from red pepper leaves to fix the leaves, experiments were conducted on the red pepper seedlings in the greenhouse. The yeast strain, which had been germinated 3 weeks after the germination, was seeded with 1 and 2 OD ₆₀₀ values of the yeast strain. The leaf suspension was sprayed to the top part of the pepper in an amount of 50 ml, and the leaf was sampled. Three pieces were placed in a 1.5 ml tube containing 1 ml of sterilized water and vortexed sufficiently. This was diluted 1/10 with the stock solution, and CFU value was measured by counting the number of colonies plated by 100 microliters on a YPD agar plate as a medium for yeast culture. And was sampled at intervals of 10 days from the spraying (Day 0) (FIG. 10). It was confirmed that there was not much difference according to the OD ₆₀₀ value sprayed at the beginning, and it was confirmed that the yeast strains which were settled on the leaf disk of 10 mm were maintained at 10 ⁴ or more from the 20th day to the 30th day. These results suggest that the isolated yeast strains live well on the leaf of the red pepper and their density is about 10 ^4, which is the most suitable growth concentration of yeast. In addition, it was analyzed that this excellent leaf coverage ability resulted in long - term induction resistance of crops to survive well on the leaves during spraying on the crops, so that they do not easily catch bacterial or viral diseases.

Korea Biotechnology Research Institute KCTC18338P 20141106

<110> Korea Research Institute of Bioscience and Biotechnology <120> Pseudozyma churashimaensis RGJ1 strain isolated from pepper plant          and uses <130> PN14326 <160> 14 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 tccgtaggtg aaccttgcgg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 tcctccgctt attgatatgc 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 cgttgccgct atctctgcta t 21 <210> 4 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ggatgctgca tactgacaaa ca 22 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 aatgaagtgg tgctcaacta caca 24 <210> 6 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 ttttggagca ttcaaccatt tgga 24 <210> 7 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 7 aagatcagac acatgga 17 <210> 8 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 8 caagcaaggg tatgcatgt 19 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 9 acagtttcca gtgccaatca 20 <210> 10 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 10 aagcgtctcg gcagttg 17 <210> 11 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 11 aactgggatt tgagaactgc cagc 24 <210> 12 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 12 atccaaggta catatagagc ttcc 24 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 13 ctccacaaga aacaaggca 19 <210> 14 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 14 gtacgaagca cgcacacaa 19

Claims (10)

( Xanthomonas axonopodis pv. Vesicatoria), cucumber mosaic virus (CMV), bovine spongiform encephalopathy virus (BBWV), pepper stain virus (PepMoV) or pepper pale stinging virus (PMMoV) isolated from pepper plants. Pseudozyma churashimaensis RGJ1 strain (KCTC18338P) inducing induced systemic resistance. delete delete A method for screening a bacteriostatic pathogenic bacterium ( Xanthomonas axonopodis pv. Vesicatoria), cucumber mosaic virus (CMV), bovine spongiform encephalitis virus (BBWV), pepper stain virus (PepMoV) or pepper plants Microorganism preparation for controlling disease caused by spot virus (PMMoV). delete A microorganism preparation for increasing the yield of a pepper plant comprising the strain of claim 1 or a culture thereof as an active ingredient. delete A method for producing a microorganism preparation comprising culturing the strain of claim 1. A method for the treatment of bacterial flora ( Xanthomonas axonopodis pv. Vesicatoria), cucumber mosaic virus (CMV), cucumber mosaic virus (CMV), and cucumber mosaic virus in a pepper plant comprising immersing the strain of claim 1 or a culture thereof in a seedling of a pepper plant, A method for controlling a disease caused by a bean curd virus (BBWV), a red pepper stain virus (PepMoV), or a red pepper stain virus (PMMoV). delete

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