KR102574692B1 - New microorganism Bacillus velezensis CMML21-47 or microbial agent comprising the same - Google Patents

Abstract

The present invention relates to the novel microorganism Bacillus velezensis CMML21-47 (KCTC19037P). The strain has excellent control efficacy against diseases of crops such as potatoes and sweet potatoes, and has the ability to produce indole-3-acetic acid, which enhances plant growth, so that it can be easily used as a variety of plant disease control agents, a plant fertilizer, etc.

Description

New microorganism Bacillus velezensis CMML21-47 or a microbial agent containing the same {New microorganism Bacillus velezensis CMML21-47 or microbial agent comprising the same}

The present invention relates to a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a microbial preparation containing the same.

In eco-friendly agriculture, useful microorganisms or antagonistic microorganisms play a large role as a possible alternative to chemical pesticides. Today's agriculture has been immersed in increasing food production by means of breed improvement, soil fertility enhancement, pest control and weed control, but the continuous use and abuse of pesticides is causing serious pollution of soil and rivers. In addition, since these pesticides remain in crops, causing problems such as toxicity, environmental pollution, and toxicity to humans and livestock, the use of pesticides is currently being strengthened against them worldwide. As concerns about such pesticides and interest in quality of life grow, recently, as a new alternative, biological control using microorganisms or plant extracts for pest control of crops has become a subject of interest.

Microbial preparations refer to products made of bacteria, fungi, viruses, etc. that are effective in controlling plant diseases or pests. In addition to pest control, areas that can be used in agriculture include soil improvement, promotion of decomposition of organic matter, promotion of nutrient/moisture absorption, and growth promotion. and herbicidal action. Biofertilizer mechanisms such as phosphate solubilization, nitrogen fixation, and production of plant growth-promoting hormones are known as ways in which useful microorganisms help plants grow. It indirectly helps plant growth by methods known as biological control mechanisms, such as inducing disease resistance. Reported relatively high populations in healthy soil and crop rhizospheres, fluorescent Pseudomonas sp. , Bacillus sp., Streptomyces sp. etc. play a role as a medium that promotes interaction with plants, and microorganisms living in the rhizosphere of crops are highly likely to be used to increase crop yield, such as effective biological control of plant diseases and pests in the process of forming a relationship with host plants. It is expected that the agricultural utilization of

Accordingly, while the inventors were conducting research on plant disease control agents in paper crops such as sweet potatoes, the new microbial Bacillus genus strain was found to be effective in controlling sweet potato major crop diseases such as creeper and black blotch, thereby completing the present invention. It became.

Republic of Korea Patent Registration No. 10-0868901 (Name of invention: Bacillus amyloliquefaciens strain and formulation containing the same, Applicant: Dong-A University Industry-University Cooperation Foundation, registration date: November 10, 2008) Republic of Korea Patent Registration No. 10-1073932 (Title of Invention: Bacillus bellegensis PB3 strain and plant disease control method using the same, Applicant: Korea Research Institute of Chemical Technology, registration date: October 10, 2011) Republic of Korea Patent Registration No. 10-0893725 (Name of invention: Bacillus thringiensis EnB2 KCTC11394BP, culture medium and microbial preparation containing the same, Applicant: ENB Co., Ltd., registration date: April 9, 2009)

An object of the present invention is to provide a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a microbial preparation containing the same.

The present invention relates to a novel microorganism Bacillus bellegensis CMML21-47 strain characterized by having a plant disease control effect.

The strain is selected from the group consisting of Fusarium oxysporum , Ceratocystis fimbriata , Rhizoctonia solani, and Sclerotinia sclerotiorum It is characterized in that it has a control effect against one or more types of pathogens.

The strain is characterized in that it has a control effect of sweet potato storage disease. The control value for the sweet potato storage disease may be 70 to 90% when stored at 25 ° C for 30 days or more.

The plant disease is characterized in that it is a disease of sweet potato.

The strain may be capable of producing proteolytic enzymes, preferably capable of producing casein-degrading enzymes.

The strain may be capable of producing siderophores.

The strain may have a plant growth promoting effect.

The strain is characterized in that it has the ability to produce indole-3-acetic acid.

The Bacillus bellegensis CMML21-47 strain includes a 16s rRNA gene represented by the nucleotide sequence of SEQ ID NO: 1 and a gyr B gene represented by the nucleotide sequence of SEQ ID NO: 2.

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The present invention provides a composition for controlling plant diseases or a microbial preparation for promoting plant growth comprising the strain or a culture medium thereof.

It is characterized in that 14 to 16 μM of indole-3-acetic acid is produced in the culture medium, preferably in the culture medium of the CMML21-47 strain.

Accordingly, the present invention relates to a microbial pesticide or microbial fertilizer comprising the strain or a culture solution thereof.

Hereinafter, the present invention will be described in more detail.

The microbial pesticide or microbial fertilizer of the present invention may be provided in a formulation selected from the group consisting of drenching agents.

In addition, the present invention comprises the steps of diluting the microbial preparation by 10 to 1000 times to obtain a diluted solution; And, it provides a method for controlling plant pathogens comprising the step of drenching the diluted solution to the control target plants.

In another aspect, the present invention comprises the steps of diluting the microbial preparation 10 to 1000 times to obtain a diluted solution; And, it is possible to provide a method for promoting plant growth comprising the step of drenching the diluted solution to the control target plants.

The medium applicable to the culture of the strain of the present invention is not particularly limited, and preferably, LB agar (Luria Bertani Agar), LB (Lysogeny Broth or Luria Bertani) TSA (Tryptic Soy Agar), TSB (Tryptic Soy Broth), It may be selected from the group consisting of NA (Nutrient Agar), NB (Nutrient Broth), NYDA (Nutrient Yeast Dextros Agar), NYDB (Nutrient Yeast Dextros Broth), KB (Kings B medium) and KA (Kings Agar B).

It provides a plant disease control agent or plant growth promoter containing the strain or its culture medium of the present invention, the strain or its culture medium may be appropriately diluted with water or a culture medium before culturing the strain according to the purpose of use.

In the present invention, the form of the microbial preparation is not particularly limited, but it is provided as a microbial pesticide or microbial fertilizer, and more preferably, it may be provided in the form of a drenching spray.

As the microbial pesticide or microbial fertilizer, the drenching agent may use a carrier or excipient used in conventional plant pesticides. Bentonite, surfactant, talc, zeolite, etc. may be preferably used as the carrier or excipient.

The strain culture solution of the present invention may be one in which the Bacillus bellegensis CMML21-47 strain was cultured at pH 4.0 to 9.0 and 15 to 45 ° C. under aerobic conditions. More preferably, it is better to culture at pH 7.0 ~ 8.0 and 30 ~ 40 ℃ under aerobic conditions. At this time, the appropriate culture time is preferably 24 to 48 hours, and the culture method may be selected from either liquid shaking culture or solid culture.

In addition, the present invention comprises the steps of diluting the microorganism preparation 10 to 1000 times (1 × 10 ^{6 ~ 8} CFU / ㎖) to obtain a diluted solution; It provides a plant pathogen control method or a plant growth promotion method comprising the step of drenching the diluted solution to the control target plant. During the irrigation spraying, the spraying amount of each dilution solution can be appropriately adjusted according to soil distribution or plant condition.

Control target plants or growth-promoting target plants of the strain of the present invention or a microbial agent containing a culture medium thereof are not particularly limited, but may preferably be sweet potatoes.

The present invention relates to a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain, wherein the strain is selected from the group consisting of Fusarium oxysporum and black spot bacteria ( Ceratocystis fimbriata ) It has a control effect against one or more pathogens and has the ability to produce indole-3-acetic acid that enhances the growth of plants, so it can be easily used as a variety of plant disease control agents and plant fertilizers. .

Figure 1 is a phylogenetic tree showing the genetic position of the selected CMML 21-47 strain, identified by 16s rRNA gene sequence (left) and gyrB gene sequence (right), and CMML21-47 is identified as Bacillus velezensis .
Figure 2 is the result of confirming the mycelial growth inhibitory effect against plant pathogens of Bacillus velezensis CMML 21-47 strain.
Figure 3 is a graph showing the optimum growth temperature of the Bacillus velezensis CMML21-47 strain.
Figure 4 is a graph showing the optimal growth pH of the Bacillus velezensis CMML21-47 strain.
Figure 5 is the result of confirming the proteolytic enzyme activity by culturing the Bacillus velezensis CMML21-47 strain on a casein plate medium.
Figure 6 is the result of confirming the sideropore production by culturing the Bacillus velezensis CMML21-47 strain in CAS blue agar medium.
7 is a result of confirming the ability of Bacillus velezensis CMML 21-47 strain to produce indole-3-acetic acid.
8 shows the analysis results of lipopeptides, which are antibacterial substances produced by the Bacillus velezensis CMML21-47 strain.
9 shows the results of an experiment in which storage disease of sweet potatoes was controlled by spraying a culture solution of the Bacillus velezensis CMML21-47 strain.
10 shows the results of an experiment in which storage disease of sweet potatoes was controlled using a volatile substance of the selected strain using a medium in which the Bacillus velezensis CMML21-47 strain was cultured.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided to sufficiently convey the spirit of the invention to those skilled in the art, so that the disclosure herein will be thorough and complete.

<Example 1. Confirmation of genetic characteristics of antagonistic microorganisms>

Antagonistic microorganisms were selected from the rhizosphere soil of healthy sweet potatoes (Naju-si, Jeollanam-do) to control major crop diseases such as creeper and black blotch on sweet potato, which is a paper crop. For the obtained rhizosphere microorganisms, antagonistic bacteria with excellent antibacterial activity were selected through replacement culture with sweet potato disease pathogens, Fusarium oxysporum and black blotch ( Ceratocystis fimbriata ). The selected antagonistic bacteria were identified by 16s rRNA sequence analysis to identify the genus name, and phylogenetic analysis was performed after classifying the species name through comparative analysis of gene sequences. In particular, the antagonistic bacteria selected in the present invention were identified by the nucleotide sequence of the gyrB gene.

As a result of the confirmation, the antagonist strain CMML21-47 was identified as Bacillus velezensis (CMML21-47, KCTC19037P) having genes of SEQ ID NO: 1 and SEQ ID NO: 2, respectively, by analyzing the 16s rRNA and gyrB genetic sequences, and on October 12, 2022 It was deposited as Bacillus velezensis CMML21-47 with accession number KCTC19037P at the Center for Biological Resources of the Korea Research Institute of Bioscience and Biotechnology. The phylogenetic classification of these strains is shown in Figure 1, and the gene sequences of SEQ ID NOs: 1 and 2 are disclosed in Tables 1 and 2.

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<Example 2. Confirmation of antibacterial activity of antagonistic microorganisms>

Selected paper crop disease control microorganism Bacillus velezensis CMML21-47, sweet potato disease Fusarium oxysporum and black blotch disease ( Ceratocystis fimbriata ), potato disease Rhizoctonia solani ( Rhizoctonia solani ) , Skelotinia skello Thiorum ( Sclerotinia sclerotiorum ) The antibacterial activity of the selected strain was verified by replacement culture.

Fusarium oxysporum, Rhizoctonia solani, Sclerotinia sclerotiorum , and Ceratocystis fimbriata strains at 25° C. for 5 days in PDA medium cultured in.

Using a cork borer with a diameter of 5 mm, the flora of pathogens was placed in the center of a disposable petri dish dispensed with PDA medium. After shaking culture of Bacillus velezensis CMM21-47 strain in TSB liquid medium, the total number of bacteria was 1Χ 10 ⁷ CFU / ml diluted uniformly with 10 μl each of each strain was applied to three parts of a disposable petri dish dispensed and cultured for 7 to 10 days in a thermostat at 25 ° C for each pathogen, and then the degree of pathogen growth was measured. Compared with the culture of Bacillus velezensis CMML21-47 together with the medium in which only the pathogen was cultured, the degree of inhibition of the growth of the pathogen was expressed as a percentage, and the experimental results are shown in FIG.

Referring to the results of Table 3 and FIG. 2 below, all of the antagonistic microorganisms have excellent antibacterial activity against the four pathogens.

pathogen Antagonist of microbial CMML21-47
Block rate (%) plant disease Fusarium oxysporum
( Fusarium oxysporum ) 56.35±0.23 creeper
(wilt disease) Raizoxtonia Solani AG-3
( Rhizoctonia solani AG-3 ) 52.76±0.18 black rot skelotinia skelotiorum
( Sclerotinia sclerotiorum ) 56.64±0.20 various sclerotia Seratocystis fimbriata
( Ceratocystis fimbriata ) 63.60±0.16 black blotch
(various black spots)

<Example 3. Confirmation of growth conditions of antagonistic microorganisms>

Next, growth conditions of microorganisms were confirmed. The Bacillus velezensis CMML21-47 strain grew well by culturing at 30 to 40 ° C for 24 to 48 hours under agitation at 100 to 200 rpm. The optimal culture temperature was 37 ° C and the optimal culture time was 48 hours. The highest incubation temperature was 45°C, and these results are shown in FIG. 3 . The minimum incubation temperature was 15°C, although not shown in the graph.

In addition, the Bacillus velezensis CMML21-47 strain grew well in the pH range of 5.0 to 9.0, preferably pH 7.0 to 8.0, most preferably pH 7.0, and cultured at least at least pH 5.0 or higher and maximum pH 9.0 or lower. it was possible 4 shows the results.

<Example 4. Enzyme production ability of antagonistic microorganisms>

In order to confirm the enzyme production ability of the Bacillus velezensis CMML21-47 strain, the strain was pre-cultured in 5 ml of TSB for 18 hours or more, and then cultured by placing 10 μl of the culture medium on each medium. For protease activity assay, casein plate medium (Casein agar; skim mik 100 g/L, agar 20 g/L) was used.

As can be seen in Figure 5, it can be confirmed that the Bacillus velezensis CMML21-47 strain has become transparent in the medium around the strain in the casein plate medium, and excellent proteolytic enzyme secretion ability can be confirmed.

<Example 5. Confirmation of siderophore production ability of antagonistic microorganisms>

Siderophores are bacterial secretions that selectively and strongly adsorb iron (Fe ³⁺ ), a limiting nutrient in soil, and have been found to inhibit the growth of pathogens by interfering with the absorption of iron by plant pathogens in the soil. . In order to confirm the siderophore-producing ability, Bacillus velezensis CMML21-47 strain was pre-cultured in 5 ml of TSB for more than 18 hours, and then 10 μl of the culture medium was added to CAS blue agae, a minimum medium containing CAS (chrome azurol S). After inoculation and incubation at 30 ° C. for 6 days, the formation of a yellow halo zone was observed.

As a result, as shown in FIG. 6, it was confirmed that the zona pellucida was formed in the Bacillus velezensis CMML21-47 strain to produce siderophores.

<Example 6. Plant growth promoter Indole-3-acetic acid (IAA) production ability of antagonistic microorganisms>

Next, it was tested whether the Bacillus velezensis CMML21-47 strain forms Indole-3-acetic Acid (IAA), a plant growth promoter necessary to determine whether it is suitable for use as a plant fertilizer, in addition to its microbial control effect.

To this end, the strain was cultured in Nutrient broth for one day, and the strain was cultured in NB medium containing L-tryptophan for 24, 48, and 72 hours. Absorbance was measured for color change. The experimental results for this are shown in FIG. 7 .

As confirmed in Figure 7, it can be seen that the selected strain, Bacillus velezensis CMML21-47 strain, produces Indole-3-acetic Acid (IAA), a factor that promotes plant growth.

<Example 7. Confirmation of antibacterial substance producing ability of antagonistic microorganisms>

It was confirmed whether the selected strain produced an antibacterial substance. To this end, analysis was performed using an ACQUITY UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm; Waters) in Acquity UPLC I-Class PLUS (Waters) using the strain culture medium as a sample.

Samples were analyzed by the gradient method at a flow rate of 0.4 ml/min using mobile phase solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile). In addition, mass spectrometry was performed using Xevo-G2- XS QTOF LC-MS (Waters) was used in positive mode in the range of 100-1600 Da. For this purpose, a capillary voltage of 3.0 kV, a source temperature of 150 ° C, and Ar were set as the collision gas MassLynx software (Waters Corporation, Milford , United States) was used to detect mass and molecular formula, and iturin, bacillomycin, fengycine, and surfactin (sigma, USA) were used as standard substances for antimicrobial substances. Based on the molecular formula, the compounds in the culture medium of each strain were identified through online and literature database searches.The analysis results are shown in FIG.

Referring to Figure 8, the main peak of the lipopeptide extract of the selected antagonistic microorganism (CMML 21-47) was detected at 7.5-13.5 minutes. Retention times of the peaks at 7.5-9.5 min, 9.5-10.5 min, and 12-13.5 min were consistent with the iturin, fengycin, and surfactin standards.

This was confirmed through MS spectrum analysis as well, that the strains contained substances of the bacillomycin, fengycin, and surfactin series, and were verified through comparison with standard substances, which were 3 types of bacillomycin, 5 types of fengycin, and 5 types of surfactin, respectively.

<Example 8. Confirmation of efficacy as a microbial control agent in field-grown plants>

Bacillus velezensis CMML21-47 culture medium and azoxystrobin, a pesticide used for actual control, were treated with sweet potatoes (Beniharuka variety) growing in the field (located in Gwangju), and then vine-pecking disease ( Fusarium oxysporum ) The control ability of was confirmed.

Specifically, the Bacillus velezensis CMML21-47 strain was added to an industrial medium (yeast extract 10 g/L, glucose 8 g/L, K ₂ HPO ₄ 2.5 g/L, NaCl 1.5 g/L, NaCO ₃ 0.5 g/L, MgSO ₄ 1 g/L, MnSO ₄ 0.3 g/L) at 30°C and 150 rpm for 24 hours with shaking, a culture diluent having a concentration of 1X10 ⁷ cfu/ml was prepared and treated individually. As a positive control group, 21.7 μg/ml of azoxystrobin, a chemical pesticide, was prepared and used. Sweet potato seedlings one month after planting were drenched twice with 100 mL each of 100 mL of culture diluent and azoxystrobin to the stem and soil of Beniharuka cultivar at 10-day intervals. After 10 days of the second treatment, Fusarium oxysporum , a vine-pecking pathogen, and Ceratocystis fimbriata , a black blotch pathogen, cultured for 7 days in a PDB medium, were developed by covering the stem and soil treated with the culture solution, and cultured 10 days after inoculation. The diluted solution was treated with irrigation in the same manner as above, and the results are shown in Table 4.

Control (%) creeper
( Fusarium oxysporum ) black blotch
( Ceratocystis fimbriata ) untreated - - Azoxystrobin 66.0±2.1 51.7±8.1 CMML21-47 strain 78.5±2.3 68.2±6.8

As can be seen in Table 4, the control value for vine pecking disease was 78.5% in the Bacillus velezensis CMML21-47 strain treatment group compared to the untreated group. The control value for black blotch was 68.2%.

Through the results of Table 4, it can be seen that the Bacillus velezensis CMML21-47 strain has a higher disease control rate than azoxystrobin used as a commercially available pesticide.

<Example 9. Confirmation of sweet potato storage disease control ability as a microorganism control agent>

Bacillus velezensis CMML21-47 strain culture solution is directly sprayed or smeared on chestnut sweet potato (Haenam) in a box of 1.5 kg, which can be purchased on the market, and stored together with a plate medium to prevent diseases occurring during storage of sweet potato and maintain quality I checked if it could be done. Sweet potatoes were stored at 25 ° C for 30 days. All experiments were conducted in triplicate.

Example 9-1. Spray treatment of culture medium

Bacillus velezensis CMML21-47 strain was cultured in TSB liquid medium at 30° C. with shaking for 18 hours or more, and then diluted to 1X10 ⁸ CFU/ml. The culture solution containing the Bacillus velezensis CMML21-47 strain was sprayed individually and evenly on the sweet potatoes with a small sprayer at 1 ml each. After treatment with the culture medium, it was put in a paper box that was included at the time of purchase, and put into a sterilized plastic box and stored at 25 ° C. for 30 days, respectively.

The results of this experiment are shown in Table 5 and FIG. 9, and compared to the control, the Bacillus velezensis CMML21-47 strain inhibited the development and progression of sweet potato storage disease by 87.9% or more. In addition, it was confirmed that the quality of the sweet potato was maintained, such as the fact that the volume reduction hardly occurred and the color change was remarkably small.

Microbial broth Control (%) untreated - CMML21-47 strain 87.9±3.1

Example 9-2. Treatment of volatiles using solid culture

Bacillus velezensis CMML21-47 was cultured in TSB liquid medium at 30 ° C. for one day with shaking, and then 200 μl was spread on TSB solid medium and cultured at 30 ° C. for more than 24 hours.

The lids of 30 petri dishes in which the strain was solid-cultured in a sterilized plastic box were opened and stored together with sweet potatoes at 25 ° C. for 30 days.

The results of the experiment are shown in Table 6 and FIG. 10.

volatile Control (%) untreated - CMML21-47 strain 72.6±6.7

As a result of confirmation, all experimental groups showed more than 70% of control value, compared to the soft and decayed ones due to the occurrence of various storage diseases in the untreated group.

As such, it was confirmed that the occurrence and progression of pathogens occurring during sweet potato storage can be inhibited only by storing the solid culture medium together.

Korea Research Institute of Bioscience and Biotechnology Biological Resources Center (KCTC) KCTC19037P 20221012

Claims (11)

As a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain having plant disease control effect of potato or sweet potato,
The plant disease is Fusarium oxysporum ( Fusarium oxysporum ) Caused by creeping vines, Seratocystis fimbriata ( Ceratocystis fimbriata ) Caused by black blotch, Rhizoctonia solani ( Rhizoctonia solani ) Caused A strain characterized by being selected from the group consisting of black spot rot and sclerotinia caused by Sclerotinia sclerotiorum . delete delete According to claim 1,
The strain is characterized in that the ability to produce proteolytic enzymes. According to claim 4,
The strain, characterized in that the protein is casein. According to claim 1,
The strain is a strain characterized in that there is a sidero pore production ability. According to claim 1,
The strain is characterized in that the growth promoting effect of potatoes or sweet potatoes. A composition for controlling plant diseases of potatoes or sweet potatoes, comprising a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a culture solution thereof,
The plant disease of the potato or sweet potato, Fusarium oxysporum ( Fusarium oxysporum ) Caused by vine splinter disease, Seratocystis fimbriata ( Ceratocystis fimbriata ) Caused by black spot disease, Rhizoctonia A composition for controlling plant diseases, characterized in that it is selected from the group consisting of black spot rot caused by solani and scleroderma caused by Sclerotinia sclerotiorum . As a microbial agent for promoting the growth of sweet potatoes or potatoes, comprising a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a culture solution thereof,
The strain is Fusarium oxysporum ( Fusarium oxysporum ) Caused by creeping vines, Seratocystis fimbriata ( Ceratocystis fimbriata ) Caused by black blotch, Rhizoctonia solani ( Rhizoctonia solani ) Caused A microbial agent characterized in that it has a control effect against plant diseases selected from the group consisting of scleroderma caused by black spot rot and sclerotinia sclerotiorum . As a microbial pesticide for potatoes or sweet potatoes, comprising a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a culture solution thereof,
The strain is Fusarium oxysporum ( Fusarium oxysporum ) Caused by creeping vines, Seratocystis fimbriata ( Ceratocystis fimbriata ) Caused by black blotch, Rhizoctonia solani ( Rhizoctonia solani ) Caused A microbial pesticide characterized in that it has a control effect against plant diseases selected from the group consisting of sclerotinia caused by black rot and sclerotinia sclerotiorum . As a microbial fertilizer for promoting the growth of potatoes or sweet potatoes, comprising a novel microorganism Bacillus velezensis CMML21-47 ( Bacillus velezensis CMML21-47, KCTC19037P) strain or a culture solution thereof,
The strain is Fusarium oxysporum ( Fusarium oxysporum ) Caused by creeping vines, Seratocystis fimbriata ( Ceratocystis fimbriata ) Caused by black blotch, Rhizoctonia solani ( Rhizoctonia solani ) Caused Microbial fertilizer characterized in that it has a control effect on plant diseases selected from the group consisting of sclerotinia caused by black rot and sclerotinia sclerotiorum .