KR20040068794A - Antifungal bacillus lentimorbus WJ5, biopesticide containing the same and mutant strain of the microorganism - Google Patents

Abstract

PURPOSE: An antifungal microorganism Bacillus lentimorbus WJ5, biological pesticide containing the same microorganism and a mutant strain of the same microorganism are provided, which microorganism has wide antifungal spectrum, so that it can be useful for control plant diseases by various fungi. CONSTITUTION: The antifungal microorganism Bacillus lentimorbus WJ5(KFCC-11316) is provided, wherein Bacillus lentimorbus WJ5(KFCC-11316) is isolated from waste net in the seashore. The biological pesticide for control of pathogenic fungi contains Bacillus lentimorbus WJ5(KFCC-11316) or Bacillus lentimorbus WJ5(KFCC-11316) cultured medium, wherein the pathogenic fungi is selected from Alternaria alternata, Alternaria solani, Botrytis cinerea, Botryoshaeria dothidea, Colletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia grisea, Phythophthora capsici, Fusarium oxysporum, Sclerotinia sclerotiorum, Mycospharella melonis and Candida albicans. A mutant strain of Bacillus lentimorbus WJ5-m12(KFCC-11317) having no antifungal activity is provided.

Description

Bacillus lentimovs ＷＪ5 having antifungal activity, microbial pesticides containing the same and mutant strains of the microorganisms {Antifungal bacillus lentimorbus WJ5, biopesticide containing the same and mutant strain of the microorganism}

The present invention relates to a strain of the genus Bacillus excellent in antifungal activity, a microbial pesticide for controlling pathogenic fungi containing the same, and a mutant strain in which antifungal activity is lost by irradiating the strain.

Organic synthetic pesticides, which are indispensable to effectively control plant diseases, are mainly manufactured by chemical synthetic methods, which not only cause extreme environmental pollution, but also have serious adverse effects on end-users by the food chain of the ecosystem. have. As an alternative to organic synthetic pesticides, the development of environmentally friendly microbial pesticides is urgently required, and the research and development of such microbial pesticides is actively progressing.

Microbial pesticides are products produced by microorganisms separated or selected from nature or modified for use in order to control pests, weeds and pathogenic microorganisms of crops, or compounds secreted by microorganisms. An essential element in the development of microbial pesticides is to select or modify useful microorganisms having excellent antagonism, and to mass produce the selected microorganisms or drugs secreted by the microorganisms and to efficiently deliver them to the crops.

Microbial metabolites involved in disease suppression through biological control methods include antifungal agents, siderophores, NH₃Enzymes, hydrogen cyanide, etc., which have antagonistic activity. Microorganisms utilized to control biologically reported fungi so far PseudomonasPseudomonas), Bacillus (Bacillus), Streptomyces (Streptomyces), Agrobacterium (Agrobacterium), Burcholderia (BurkholderiaBacteria and trichoderma ()Trichoderma), Ziroclidium (Gilocladium), Fusarium (FusariumFungi such as (Fravel, D. R., W. J. Connick, and J. A. Lewis.,Kluwer Academic Pubishers, Dordrecht, London, 1998, 187-202).

Korean Patent Registration No. 6238 (new Bacillus subtilis subspecies and antifungal substance KRF-001 complexes produced therefrom) contains Bacillus subtilis subsplices that produce antifungal oligopeptide-based substances. Krictiensis ), but the antifungal activity spectrum has a narrow problem. In addition, Korean Patent Application No. 193-22037 (Method for producing microbial insecticide using bioencapsulation), a natural gel matrix rich in water is mixed with water to prepare a protein-rich natural gel matrix, and autoclaved to Bacillus terringiensis spores There is known a method for preparing a bioencapsulated microbial insecticide by mixing and drying the same, but there is a problem that the antifungal activity spectrum is narrow and the components and composition of the biomatrix are complicated. Korean Patent Application No. 1998-27468 (Improved Coating Pesticide Matrix, Preparation Method and Composition Containing It) is applied to a coated pesticide matrix comprising a pesticide, a polymer, a plasticizer, a sunscreen, an activity enhancer, and a suspending agent. Although the contents are known, there are problems such as narrow antifungal activity spectrum and limited effects.

In addition, until now, many developments have been attempted to directly use the isolated functional strains or directly use the production products of the strains, but attempts to isolate and apply mutants whose functions have been changed from these isolates, in particular, to prepare mutants using gamma rays. There is almost no situation.

Accordingly, the inventors of the present invention confirmed that Bacillus lentimovs, which is isolated from the pulmonary net, has excellent activity against various pathogenic fungi and has resistance to pesticides while developing new microbial pesticides having excellent antifungal activity. The present invention was completed by investigating gamma rays to the strains to prepare a mutant strain that lost antifungal activity and confirming that it can be used as a functional strain.

An object of the present invention is Bacillus lentimovs WJ5 having antifungal activity against pathogenic fungi, microbial pesticides for controlling pathogenic fungi containing the same, mutant strains that lose antifungal activity by irradiating the strain and producing the mutant strains To provide a way.

1 is a photograph showing the antifungal activity spectrum of Bacillus lentimovs WJ5,

A: Alternaria Alternata, B: Alternaria Solani,

C: Botrytis Ceria, D: Botryoseria Tochdea,

E: choletotribone gloospoidoids, F: lysatonia solani,

G: Calcium Ultimum, H: Pyrricularia griacia,

I: Pytoptora capsaish, J: Fusarium oxysporium,

K: sclerotinia sclerotirium, L: mycosparella melonis

2 is a photograph showing antifungal activity against Candida albicans of Bacillus lentimovs WJ5,

control: a paper disk containing no strain,

WJ5: Bacillus Lentimovs WJ5,

3 is a graph showing the sensitivity according to gamma dose irradiated to Bacillus lentimovs WJ5.

In order to achieve the above object, the present invention provides Bacillus lentimovs WJ5 having antifungal activity against pathogenic fungi.

The present invention also provides a microbial pesticide for controlling pathogenic fungi containing the microorganism.

In addition, the present invention provides a mutant strain in which antifungal activity is lost by irradiating the microorganisms with radiation.

In addition, the present invention comprises the steps of 1) recovering the cells survived after irradiation with Bacillus lentimovs WJ5 showing a mortality of about 95%;

2) selecting a strain having reduced antifungal activity against pathogenic fungi among the cells surviving in step 1;

3) recovering the surviving cells after irradiating the strain selected in step 2 again with radiation showing a mortality of about 95%;

4) selecting a strain that has lost antifungal activity against pathogenic fungi from the cells recovered in step 3;

5) It provides a method for producing the mutant strain comprising the step of screening the mutant strain does not occur return mutation after subcultured strain selected in step 4.

Hereinafter, the present invention will be described in detail.

The present invention provides Bacillus lentimorbus WJ5 having antifungal activity against pathogenic fungi.

The microorganisms of the present invention are microorganisms isolated from the pulmonary nets of the coast and have antifungal activity specifically against pathogenic fungi, and have excellent resistance to pesticides such as fungicides, insecticides and herbicides.

The microorganism of the present invention was cultivated Gram stain by pure culture of the selected bacteria, was identified using the fatty acid composition analysis and database of cells, and the biochemical characteristics were identified to complement the identification results ( Table 1 and Table 1 ). 2 ). As a result, the present inventors determined that the microorganism was a Bacillus lentimovs strain and named it "Bacillus lentimovs WJ5", and deposited it on Korea microbial conservation center on November 19, 2002 (Accession No .: KFCC-11316). ).

The present invention also provides a microbial pesticide for controlling pathogenic fungi comprising the microorganism.

The microbial pesticide of the present invention is characterized by containing Bacillus lentimovs WJ5 and / or its culture, and may further include other microorganisms having antifungal activity and resistance to pesticides.

Pathogenic fungi to which the microbial pesticides of the present invention can be applied are Alternaria alternata , Alternaria solani , Botrytis cinerea , Botryotinia torch , which are plant pathogenic fungi. Botryoshaeria dothidea , Colletotrichum gloeosporioides , Rhizoctonia solani , Pythium ultimum , Pyricularia grisea , Pytopia Phythophthora capsici , Fusarium oxysporum , Sclerotinia sclerotiorum and Mycospharella melonis and Candida albicans, animal pathogenic fungi ( Candida albicans ) is preferably at least one selected from the group consisting of.

In addition, Bacillus lentimovs WJ5 contained in the microbial pesticide of the present invention has resistance to pesticides currently used. Pesticides resistant to the microorganism of the present invention is a bactericide beomyl, flusilazole, oxadixyl, edifenphos, mancozeb, azocystrobin, Isoprothiorane and iprobenphos, insecticides chlorfenapyr, imidacloprid, carbofuran, fenpyroximate, chlorpyrifos and chlorpyrifos and It is preferably at least one selected from the group consisting of diazinon, butachlor, the herbicide.

The microbial pesticide of the present invention may be prepared by mixing a microbial carrier with Bacillus lentimovs WJ5, and the microbial carrier may be selected from the group consisting of plant resource polymer materials, microbial origin biopolymer materials, nutrients, auxiliary materials and additives. . Plant-based polymer materials include potato flour, soy flour and starch powder, microbial origin biopolymers include mucopolysaccharides, poly-betahydroxy-alkanoates and flocculants, and nutrients such as glucose, peptone and inorganic salts, and auxiliary materials. Is vermiculite and rice husk powder, additives include extenders, UV blockers and surfactants.

The microbial pesticide of the present invention is concentrated by culturing Bacillus lentimovs WJ5 and then filmed with mucopolysaccharide, and then sterilized and cooled by mixing the components contained in the microbial carrier, and then encapsulated Bacillus lentimovs WJ5 Mixed with and dried to prepare a powder of the appropriate size. Bacillus lentimovs WJ5 mixed with the microbial carrier is preferably 10 ⁸ -10 ⁹ cells per gram of the microbial carrier.

The microbial pesticide of the present invention is an environmentally friendly multifunctional antifungal agent which is useful for environmental conservation and purification because of its excellent antifungal activity and without the use of chemicals. It can be usefully used as a composting agent, foliar spray or irrigation spray.

In addition, the present invention provides a mutant strain in which antifungal activity is lost by irradiating the microorganisms with radiation.

The mutant strain of the present invention was selected as a strain having a different antifungal activity from the parental strain Bacillus lentimovs WJ5 among the surviving strain by irradiation with 95% lethal radiation to the Bacillus lentimovs WJ5 strain.

In the above, the radiation is preferably selected from the group consisting of gamma rays, ultraviolet rays, X-rays and E-beams, and more preferably gamma rays. In a preferred embodiment of the present invention, ⁶⁰ Co was used as the gamma source.

The mutant strain of the present invention is characterized by the loss of antifungal activity and resistance to microbial pesticides as compared to the parent strain Bacillus lentimovs WJ5 strain. The present inventors named the microorganism Bacillus lentimovs WJ5-m12 and deposited it at Korea Microorganism Conservation Center on November 19, 2002 (Accession No .: KFCC-11317).

The mutant strain of the present invention has lost antifungal activity compared to Bacillus lentimovs WJ5 strain, and since the gene is mutated, it can be usefully used for the search for antifungal activity related gene sources and substances.

The present invention also provides a method for preparing the mutant strain.

Method for preparing a mutant strain of the present invention

1) irradiating Bacillus lentimovs WJ5 with radiation having a mortality of about 95% and recovering the viable cells;

2) selecting a strain having reduced antifungal activity against pathogenic fungi among the cells surviving in step 1;

3) recovering the surviving cells after irradiating the strain selected in step 2 again with radiation showing a mortality of about 95%;

4) selecting a strain that has lost antifungal activity against pathogenic fungi from the cells recovered in step 3;

5) selecting the mutant strain that does not cause a reverse mutation after passage of the strain selected in step 4.

In the above, the radiation is preferably selected from the group consisting of gamma rays, ultraviolet rays, X-rays and E-beams, more preferably gamma rays. It is preferable to use ⁶⁰ Co as a gamma source, but it is not necessarily limited to this.

The above pathogenic fungi include Alternaria alternata, Alternaria solani, Botrytis ceria, Botryoseria dochdeea, Colletotribe gloosporoid, Lysatonia solanie, Ps. choose from wool timeom flute Temecula Liao draw Asia, Xi Shi Pai Top Torah cap, Fu saryum oxy Spokane volume, switch Klee Loti Nias Klee as thiosulfate volumes and Maiko spa Relais Melo Nice and animal pathogenic fungus Candida albicans throw the group consisting of It is desirable to be. In a preferred embodiment of the present invention, Candida albicans was used as a screening strain in step 2.

Hereinafter, it demonstrates in detail by an Example.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Isolation and Identification of Bacillus lentimovs WJ5 Strain, an Antifungal Active Bacteria

To find new microorganisms with antifungal activity, samples were collected from waste nets, and the samples were collected by dilution plate method using NA medium (nutrient agar medium, Difco) and PDA medium (potato dextrose agar medium, Difco). Strains were isolated, and among them, strains excellent in antifungal activity were isolated. Active strains were selected first through the activity test against Candida albicans using the isolated strains, and the final strain was selected by antifungal activity test against various phytopathogenic bacteria using the selected strains.

Gram staining was performed by pure culture of the selected bacteria, and identified using fatty acid methyl esters (FAME) analysis and a database using a MIDI system. Applied to API 50 CHB system), the identification results were supplemented.

Cell Fatty Acid Analysis fatty acid content(%) C13: 0 ISO 0.28 C14: 0 ISO 1.24 C14: 0 1.47 C15: 0 ISO 21.92 C15: 0 ANTEISO 38.20 C15: 0 0.68 C16: 0 ISO 3.24 C16: 1 w11c 0.86 C16: 0 11.67 ISO C17: 1 w10c 0.38 C17: 0 ISO 9.82 C17: 0 ANTEISO 9.07 C18: 3 w6c (6,9,12) 0.82 C18: 0 0.35

Carbohydrate Fermentation Analysis carbohydrate Bacillus lentibs WJ5 Bacillus lentimovs WJ5-m12 carbohydrate Bacillus lentibs WJ5 Bacillus lentimovs WJ5-m12 Control - - Esculandrolissis - - Glycerol + + Salinity + + Erythritol - - Cellobiose + + D-Arabinose - - Maltose + + L-Arabinose + + Lactose + + Ribose + - Melibiose + + D-Xylose + + Sucrose + + L-Xylose - - Trehalose + + Adonitol - - Inulin - - β-methyl-D-giloside - - Melezitos - - Galactose - + Raffinos + + Glucose + + Starch + + Fructose + + Glycogen + + Mannos + + Xylitol - - Sobos - - Genthiobios - - Rhamnos - - D-Turanos - - Dulsitol - - D-Rizos - - Inositol + + D-tagatose - - Mannitol + + D-fucose - - Sorbitol - - L-fucose - - α-methyl-D-mannoside - - D-Arabitol - - α-methyl-D-glucoside + + L-Arabitol - - N-acetyl-glucosamine - - Gluconate - - Amigdalin - - 2-keto-gluconate - - Arbutin + - 5-keto-gluconate - -

In the above +: fermentation,-: non-fermentation

Through cellular fatty acid composition analysis ( Table 1 ) and carbohydrate fermentation analysis ( Table 2 ), the microorganism of the present invention was identified as Bacillus lentimovs, the microorganism was named as Bacillus lentimovs WJ5 strain and as of November 19, 2002 Deposited to Korea Microbial Conservation Center (Accession No .: KFCC-11316).

Example 2 Antifungal Activity Spectrum of Bacillus lentimovs WJ5

The antifungal activity of Bacillus lentimovs WJ5 of the present invention was analyzed. Specifically, in order to analyze the antifungal activity spectrum, pathogenic fungi are inoculated in the center of the PDA medium in the form of agar plugs, and then the culture medium of the precultured antifungal bacteria is antagonized in vivo by the method of disk inoculation. The test was performed. The relative antifungal activity was investigated by measuring the length from the edge of the disk to the inhibition zone. The antifungal activity of the veterinary pathogenic fungus Candida albicans was prepared by mixing Candida albicans preculture with NB medium supplemented with 0.7% agar to make top agar in NA medium. 50 μl of the antifungal bacterial culture was inoculated and incubated at 37 ° C., and the antifungal activity was determined by measuring the size of the transparent inhibitor.

As a result, the Bacillus lentimovs WJ5 of the present invention is a plant pathogenic fungi Alternaria alternata , Alternaria solani , Botrytis cinerea , Botryoseria invert Botryoshaeria dothidea , Colletotrichum gloeosporioides , Rhizoctonia solani , Pythium ultimum , Pyricularia grisea , Pytopia Phythophthora capsici , Fusarium oxysporum , Sclerotinia sclerotiorum and Mycospharella melonis ( FIG. 1 ) and animal pathogenic fungi Growth inhibitory activity was shown for the Candida albicans ( FIG. 2 ).

Example 3 Preparation of a Mutant Strain of Bacillus lentimovs WJ5

Mutant strains were prepared by irradiating gamma rays to the Bacillus lentimovs WJ5 strain. Specifically, 1 mL of Bacillus lentimovs WJ5 strain, which confirmed the excellent antifungal activity in Examples 1 and 2, was placed in a microcentrifuge tube and irradiated with gamma rays of 0.05-25 kGy, followed by NA medium. The plate was incubated for 24 hours at 37 ℃. The susceptibility was investigated by the number of viable cells of each gamma ray dose, and the cells survived at a radiation dose of 13.5 kGy with a mortality of 95%. The surviving cells were inoculated in NA medium previously coated with Candida albicans and cultured for 8 hours, and then, the characteristic strains showing changes in morphological characteristics and antifungal activity were selected as mutation candidate strains. The primary use of Candida albicans in the selection of strains showing changes in antifungal activity is the rapid growth of these strains. Generally, antifungal activity against them results in activity against one or more phytopathogenic fungi. Therefore, it was used. After incubating the selected strains in liquid, strain cultures were inoculated into discs (paper disc, Advantec, Toyo) at 50 μl each and incubated at 37 ° C. for 1 day. After the incubation, the gamma ray of 13.5 kGy, which is 95% lethal dose, was irradiated, and the viable cells were cultured in NA medium previously coated with Candida albicans, and the antifungal activity against Candida albicans was compared with that of the parent strain Bacillus lentimovs WJ5. Mutant strains that were not passaged back were selected as mutant strains by passing the missing strains four times.

As a result, when the Bacillus lentimovs WJ5 of the present invention was irradiated with radiation, the D ₁₀ value, which is an indicator of radiation sensitivity, was found to be 2.57 kGy ( FIG. 3 ), and the mutant strain prepared therefrom was the parent strain Bacillus. Compared to lentimovs WJ5, the activity against most fungi except Mycosparella melonis was lost ( Table 3 ).

Phytopathogenic fungi Bacillus lentimovs WJ5 Bacillus lentimovs WJ5-m12 Alternaria Alternata + - Alternaria Solani + - Botrytis Cinema + - Botryoseria Tochdea + - Colletotribu Glueosepoid + - Raiatonia Solani + - Calcium Ultimum + + Ficiraria Grisia + - Pytoptora capsaishi + - Fusarium Oxysporium + - Sclerotinia sclerotirium + - Mycosparella Melonis + -

-: Inactive, +: active

The present inventors named the mutant strain Bacillus lentimovs WJ5-m12 and deposited it with Korea Microorganism Conservation Center on November 19, 2002 (Accession No .: KFCC-11317).

Example 4 Analysis of Resistance and Growth Capacity of Commercial Bacillus Bacillus Lentimovs WJ5 and Bacillus Lentimovs WJ5-m12

The resistance and growth capacity of Bacillus lentimovs WJ5 and Bacillus lentimovs WJ5-m12 of the present invention against commercial pesticides contaminated with existing cropland were investigated. Specifically, by investigating pesticides used most frequently in arable land, we selected 20 pesticides including 11 fungicides, 7 pesticides, and 2 herbicides. It was used according to the distinction. The general names and uses of these pesticides are shown in Table 4 , and the resistance to pesticides was determined by inoculating 50 μl of the bacterial culture cultured in the NA medium to which the concentration of the pesticide used in the farmland was added. The growth was measured by culturing daily.

Type of pesticide Common name water(%) Concentration (ppm) menstruum Fungicide Benomyl 95.0 250 Acetone Flusilazole 98.6 20 Methanol Tebuconazole 98.0 250 Isopropanol Oxadixyl 98.0 250 Methanol Edifenphos 95.0 300 Methanol Mancozeb 83.0 1,000 H ₂ O Pencycuron 97.0 250 Isopropanol Azoxystrobin 98.4 100 Acetone Copper hydroxide 89.0 800 ammonia Isoprothiorane 96.0 400 H ₂ O Iprobenphos 97.0 50 Acetone Insecticide Chlorfenapyr 93.2 50 Acetone Imidacloprid 95.0 50 Isopropanol Carbofuran 75.0 300 Isopropanol Penpyroximate 98.0 25 Methanol Chlorpyrifos 93.0 200 Methanol BPMC 93.0 500 Acetone Diazinon 95.0 30 Acetone Herbicide Butachlor 87.9 50 Acetone Glyphosate 98.0 50 H ₂ O

As a result, Bacillus lentimovs WJ5 is Bennoyl, Flusilazole, Oxadixyl, Edifenphos, Mancozeb, Azoxystrobin, Isoprothio Isoprothiorane, Iprobenphos, Chlorfenapyr, Imidacloprid, Carbofuran, Penpyroximate, Chlorpyrifos, Diazinon And it showed excellent resistance to butachlor (Butachlor) showed high growth in the pesticide-added solid medium, mutant strain Bacillus lentimovs WJ5-m12 also showed the same resistance to pesticides as the parent strain ( Table 5 ).

Type of pesticide Bacillus lentimovs WJ5 Bacillus lentimovs WJ5-m12 Benomyl + + Flusilazole + + Tebuconazole - - Oxadixyl + + Edifenphos + + Mancozeb + + Pencycuron - - Azoxystrobin + + Copper hydroxide - - Isoprothiorane + + Iprobenphos + + Chlorfenapyr + + Imidacloprid + + Carbofuran + + Penpyroximate + + Chlorpyrifos + + BPMC - - Diazinon + + Butachlor + +

-: No resistance; +: Resistant.

Example 5 Analysis of Organic Resolution of Bacillus lentimovs WJ5 and Bacillus lentimovs WJ5-m12

The degradation ability of the Bacillus lentimovs WJ5 and Bacillus lentimovs WJ5-m12 of the present invention to fatty acids, starch, cellulose and lignin were analyzed. Specifically, the lipidolytic activity is described in Sierra, G., 1957, A simple method for the detection of lipolytic activity of microoganisms and some observations on the influence of the contact between cells and fatty substrates, Antonie van Leeuwenhoek, 23, 15-22.) Was strained to inoculate the strain into a solid medium containing tween-80 (10 g of peptone, 0.1 g of CaCl ₂ , 15 g of agar, 1 ml / l of tween-80). After incubation at 25 ° C. for 3 days, when opaque crystals appeared around the cells, they were considered to have fatty acid degradation capacity. Amylase activity was determined by medium containing soluble starch (starch 5 g, peptone 10 g, CaCl ₂ 0.5 g, MgCl ₂ 5 g, MgSO ₄ 2 g, KCl 1 g, FeSO ₄ 0.001 g, Agar was incubated at 25 ° C. for 1.5 days at 1.5% / L), and then gram iodine (Gram's iodine) solution was added to the cells to observe the formation of a transparent inhibitor around the cells. Cellulolytic activity was measured at 25 ° C. in carboxymethylcellulose (CMC) medium (CMC 5 g, MgSO ₄ 2 g, CaCl ₂ 0.5 g, KCl 1 g, FeSO ₄ 0.001 g, agar 1.5% / l). After incubation for 5 days, 0.5% Congo-red (Congo-red) solution was measured by treating the cells with the presence of a zona pellucida. Ligninase activity was determined by adding poly-R (poly R-478 dye, Sigma) to medium (KH ₂ PO ₄ 0.6 g, MgSO ₄ 7H ₂ O 0.5 g, K ₂ HPO ₄ 0.4 g, (NH ₄ ) ₂ tartrate 0.22 g, sorbose 4 g, poly-R 0.2 g, mineral solution (mineral soln: CaCl ₂ 2H ₂ O 7.4 g, ferric citrate 1.2 g, ZnSO ₄ 7H ₂ O 0.7 g , MnSO ₄ 5H ₂ O, CoCl ₂ 6H ₂ O 0.1 g, Thiamine-HCl 10 mg / L) 10 ml, agar 1.5%, pH 4.5) incubated at 25 ℃ for 5 days, the presence of a zona pellucida around the cells It was measured by observing (Glenn, JK and Gold, MH 1983, Appl. Environ. Microbiol ., 45, 1741-1747).

As a result, Bacillus lentimovs WJ5 and Bacillus lentimovs WJ5-m12 showed no activity on lipolysis, fibrinolytic, and lignin, but showed starch degrading activity and organic matter degrading ability. Table 6 ).

Strain Enzyme activity ^a Lipolytic enzymes Starch degrading enzyme Cellulose degrading enzyme Lignin degrading enzyme Bacillus lentimovs WJ5 - + - - Bacillus lentimovs WJ5-m12 - + - -

^a enzyme activity; -: No enzyme activity, +: enzyme activity.

As described above, the Bacillus lentimovs WJ5 of the present invention exhibits a broad antifungal spectrum and is effective in controlling plant diseases caused by various fungi, as well as having high resistance and viability to commercial chemical pesticides. It can be used as, and also has the ability to decompose organic matter can be used as a multifunctional environmentally friendly soil improver, composting agent, environmental purifier. In addition, Bacillus lentimovs WJ5-m12 which has lost antifungal activity by gamma irradiation to the microorganism can be useful for the search for genes and substances related to antifungal activity.

Claims (11)

Bacillus lentimorbus WJ5 having antifungal activity against pathogenic fungi (Accession No .: KFCC-11316). A microbial pesticide for controlling pathogenic fungi, comprising the microorganism of claim 1 and / or a culture solution thereof. 3. The pathogenic fungus according to claim 2, wherein the pathogenic fungi are Alternaria alternata, Alternaria solani, Botrytis cieria, Botryoseria dochdeea, Choletotribe gloospoidoid, Lysatonia solani, Psium wool timeom, that at least one of which flutes Temecula Liao draw Asia, pies saw Fu saryum, at between Torah cap oxy Spokane volume, switch Klee Loti Nias Klee to choose from Tio volume, Maiko spa Relais Melo Nice and Candida albicans throw the group consisting of Characterized by microbial pesticides. 3. The microbial pesticide of claim 2, further comprising a microbial carrier in the microorganism and / or culture medium of claim 1. 5. The microbial pesticide of claim 4, wherein the microbial carrier is selected from the group consisting of plant resource polymeric materials, microbial origin biopolymer materials, nutrients, auxiliary materials and additives. The Bacillus lentimovs mutant strain of which antifungal activity is lost by irradiating the strain of claim 1. 7. The Bacillus lentimovs WJ5-m12 mutant strain (Accession No .: KFCC-11317) according to claim 6, wherein the antifungal activity against Candida albicans has been lost. 1) recovering the viable cells after irradiating the strain of claim 1 with a radiation rate of about 95%; 2) selecting a strain having reduced antifungal activity against pathogenic fungi in the viable cells in step 1; 3) recovering the surviving cells after irradiating the strain selected in step 2 again with radiation showing a mortality of about 95%; 4) selecting a strain that has lost antifungal activity against pathogenic fungi from the cells recovered in step 3; And 5) A method for preparing the strain of claim 6, comprising the step of selecting the mutant strain that does not occur a reverse mutation after passage of the strain selected in step 4. The method of claim 8, wherein the radiation is selected from the group consisting of gamma rays, ultraviolet rays, X-rays, and E-beams. 10. The method of claim 9, wherein the radiation is gamma rays. The method of claim 8, wherein the pathogenic fungi are plant pathogenic fungi Alternaria alternata, Alternaria solani, Botrytis ceria, Botryoseria dochdeea, Choletotribe gloeosporoid, Lysatonia solani, pisyum wool timeom flute Temecula Liao draw Asia, pies Top Tora Fu saryum, when between the cap oxy Spokane volume, switch Klee Loti Nias Klee as thiosulfate volumes and Maiko spa Relais Melo Nice and animal pathogenic fungus Candida albicans throw And selected from the group consisting of: