KR101977173B1 - Chromobacterium sp. JH7 strain having antifungal activity against plant pathogen and uses thereof - Google Patents

Abstract

The present invention relates to a JH7 strain in the genus Chromobacterium having antifungal activities against causative pathogens of plant diseases, and a usage thereof. The JH7 (KACC92211P) strain in the genus Chromobacterium having antifungal activities against causative pathogens of 11 plant diseases including Cylindrocarpon destructans, a causative pathogen of ginseng root rot disease is harmless to a human body, does not cause environmental pollution and has excellent control activities on causative pathogens of various plant diseases. Accordingly, the present invention is a biological pesticide for controlling plant diseases to be able to replace a chemical pesticide, and is able to be provided as a breakthrough biological control agent to prevent problems caused by environmental pollution.

Description

Chromobacterium genus JH7 strain having antifungal activity against plant pathogenic bacteria and its use {Chromobacterium sp. JH7 strain having antifungal activity against plant pathogen and uses thereof.

The present invention relates to a strain of Chromobacterium genus JH7 having an antifungal activity against plant pathogenic bacteria and its use.

Panax ginseng is generally used in the treatment of a variety of diseases, including traditional medicine and cancer and diabetes. Ginseng is a perennial medicinal plant that grows in the shade. It takes 4 to 6 years to produce mature roots, which are also used as raw materials for producing red ginseng. Ginseng is damaged by various soil microorganisms including fungi, bacteria and nematodes by long-term cultivation and shade cultivation environment. Among these microorganisms, they are seriously damaged by plant pathogenic fungi during cultivation of ginseng. For example, Colletotrichum gloeosporioides is an anthrax, Alternaria panax is Alteraria wilt , Ascochyta sp., And Macrophoma sp. ), Botrytis cinerea causing gray fungal disease ( Botrytis cinerea ), Cladosporium spp. is a dry rot disease, Phytophthora ( Phytophthora sp. cactorum ) is a phytophthora blight, Pythium ultimum and Rhizoctonia solani cause damping off and Sclerotinia panacicola causes sclerotinia rot. Among the phytopathogenic fungal pathogens affecting ginseng, Cylindrocarpon destructans = Nectria radicicola ) caused by root rot rot disease. Soil pathogenic microorganism, Shirindrocapone desatuntense, causes root rot in ginseng causing economic loss of 40 ~ 60% production of ginseng. In addition, since it can survive in soil for more than 10 years after ginseng harvest, it is highly likely to recur if soil management is insufficient.

The use of chemical disinfectants to control ginseng root rot disease improves the productivity of ginseng, but it is accompanied by toxic side effects such as residual toxicity, overuse, and destruction of ecosystem. Recently, it has been demanded to develop environmentally friendly control technology. Typically, it is a method using secondary metabolites using microorganisms that are easily degraded in nature and have selectivity for pathogens. Biological control mechanisms that control plant diseases through microorganisms fall into five categories: 1) lytic action that destroys the cell walls of phytopathogenic fungi; 2) antimicrobial substances produced in the genus Bacillus, penicillin, Pseudomonas and Streptomyces; 3) competitive antagonism that inhibits the growth and propagation of pathogenic bacteria by competing with the parasitic action of fungal pathogens in plants; 4) inhibition of fungal pathogenicity of plants and depletion of elements necessary for growth, such as nutrients in the rhizosphere; Inducible resistance which induces tolerance to diseases by activating plant immune function through lipopolysaccharide, salicylic acid, hydrogen cyanide and 2,3-butanediol produced by microorganisms.

Natural antibiotics derived from microorganisms provide important materials for the development of new disinfectants, and various antimicrobial active ingredients are of great value in antibiotic research. Studies on biological control using antagonistic microorganisms in ginseng root rot disease have been carried out by plant growth promoting rhizobacteria such as Bacillus spp. And Streptomyces spp. In the present invention, microorganisms that exhibit antagonistic action against Shirindokone death tracts in the ginseng rhizosphere soil were isolated and identified. And we investigated the production of siderophores, including cellulase, protease, and chitinase, and the ability of insoluble phosphoric acid to decompose by microbial secondary metabolite activity. In addition, mycelial growth inhibition, spore and germination inhibition, and suppression of thick membrane spore formation were investigated in selected microorganisms.

Korean Patent No. 0757300 discloses a strain C-61 strain of the genus Chromobacterium having plant disease-controlling activity and a plant herbal preparation using the same. In Korean Patent No. 1717137, Bacillus subtilis SCHB1433 strain having an ability to secrete an extracellular enzyme and having an antifungal activity against plant pathogenic bacteria, and its use ", as disclosed in the present invention, ≪ RTI ID = 0.0 > JH7 < / RTI >

The present invention has been made in view of the above-mentioned needs. In the present invention, the JH7 strain having excellent antimicrobial activity against Cylindrocarpon destructans , which is a germ rot root rot germ, was isolated from soil, It was confirmed to be a bacterium-like strain. In addition, it was confirmed that the above-mentioned Chromobacterium genus JH7 has antifungal activity against eleven kinds of fungi including Shilindrocarpane desaturnton. In addition, it has been confirmed that chitinanase and protease releasing ability, Phosphoric acid decomposition ability, spore formation inhibiting ability, spore germination inhibiting ability and thick membrane spore formation inhibiting ability, and thus it can be used as a biocide pesticide harmless to the human body.

In order to solve the above problems, the present invention provides a strain of Chromobacterium genus JH7 having an antifungal activity against plant pathogenic bacteria.

The present invention also provides a plant disease controlling composition comprising the strain, a culture solution thereof or a culture solution extract thereof as an active ingredient.

The present invention also provides a method of controlling a plant disease comprising the step of treating an effective amount of the plant disease control composition to a plant part, soil or seed.

The chromosuberitic JH7 strain (KACC92211P), which has antifungal activity against eleven plant pathogenic bacteria including Cylindrocarpon destructans , which is the causative agent of ginseng root rot of the present invention, is harmless to the human body and causes environmental pollution And it was confirmed that it exhibited excellent controlling activity against various plant pathogenic bacteria. Therefore, the present invention can provide a bio-pesticide for plant disease control, which can replace chemical pesticides and can be used as a novel biological control agent to prevent problems caused by environmental pollution.

Figure 1 shows the result of investigation of the antibacterial activity of JH7 strain. (A) 10 days after the inoculation, the cells were cultured in a PDA medium with Cylindrocarpon destructans ); Bacteria treated (left), control (right). (B) Colony type (24 hpi) of JH7 strain cultured in LB medium.
Fig. 2 shows the results of the systematic similarity analysis based on the 16S rRNA nucleotide sequence of the JH7 strain having antibacterial activity.
FIG. 3 shows the enzyme activity, cytidophor production and insoluble phosphoric acid solubilization of JH7 strain.
FIG. 4 shows the results of inhibition of growth of Cylindrocarpon destructans by the strain JH7 of the genus Chromobacterium. (A) Shirindrocapone death spore production, (B) assay for germination inhibition effect, (C) inhibition of thick membrane spore formation assay

In order to achieve the object of the present invention, the present invention provides a Chromobacterium sp. JH7 strain having an antifungal activity against plant pathogenic bacteria.

In the present invention, a JH7 strain having excellent antimicrobial activity against Cylindrocarpon destructans , which is a genus of ginseng root rot disease, was isolated from the soil and identified as Chromobacterium sp. And deposited on November 16, 2017 (National Institute of Agricultural Science and Technology) as the strain JH7 of chromobacterium (Accession No .: KACC92211P)

In addition, it was confirmed that the above-mentioned Chromobacterium genus JH7 has antifungal activity against eleven kinds of fungi including Shilindrocarpane desaturnton. In addition, it has been confirmed that chitinanase and protease releasing ability, Phosphoric acid degradation ability, spore formation inhibitory ability, spore germination inhibitory ability and chlamydospore formation inhibitory ability were confirmed.

In a strain according to an embodiment of the present invention, the fungus is selected from the group consisting of Dothideomycetes , Agaricomycetes , Leotiomycetes and Sordariomycetes And preferably a Cylindrocarpon ( Cylindrocarpon) destructans , Cladosporium cucumerinum , Exserohilum turcicum , Cochliobolus heterostrophus , Rhizoctonia solani , Borotitis cineum , Botrytis cinerea ), Sclerotinia sclerotiorum , Magnaporthe oryzae), Collet sat tree glutamicum Aqua tatum (Colletotrichum acutatum), Fusarium oxy sports rooms (Fusarium oxysporum and Fusarium solani , and most preferably Cylindrocarpon destructans , but is not limited thereto.

The strain according to one embodiment of the present invention may have a kitsinase and protease secretion ability, a seaward pore production ability, an insoluble phosphate decomposition ability, a spore formation inhibiting ability, a spore germination inhibiting ability and a thick spore formation inhibiting ability.

The spore is preferably selected from the group consisting of Cylindrocarpon destructans , Cladosporium < RTI ID = 0.0 > cucumerinum , Exoserohilum turcicum , Cochliobolus heterostrophus , Rhizoctonia solani , Botrytis spp. cinerea ), Sclerotinia sclerotiorum , Magnaporthe oryzae , Colletotrichum acutatum), Fusarium oxy Spokane Room (Fusarium oxysporum ) or spores produced by Fusarium solani , but are not limited thereto.

The present invention also provides a plant disease controlling composition comprising the strain, a culture solution thereof or a culture solution extract thereof as an active ingredient.

In the composition according to an embodiment of the present invention, the plant disease may be a fungal disease-causing plant disease, preferably a Cylindrocarpon destructans , Cladosporium cucumerinum , Exserohilum turcicum , Cochliobolus heterostrophus , Rhizoctonia solani , Borotitis cineum , For example, Botrytis cinerea , Sclerotinia sclerotiorum , Magnaporthe oryzae , Colletotrichum acutatum , Fusarium oxysporum or It may be a plant disease caused by Fusarium solani , and most preferably it may be, but is not limited to, ginseng roots rot caused by Shirindo carpal death tractant.

Preferably, the plant disease controlling composition may be, but not limited to, a liquid wettable powder (SC), a liquid suspension (SM), a adsorbent granule (adsorptive GR), a granular granule (prefabricated GR) or a powdery wettable powder .

The composition for controlling plant diseases includes a culture solution prepared by culturing the strain of the present invention, and can be used for an antimicrobial microbial pesticide, a seed coating agent, a microbial nutrient, a soil improving agent, a composting homework, a leaf spreading agent,

In the composition for controlling a plant disease of the present invention, the above-mentioned Chromobacterium genus JH7 strain or its culture can be used in various forms by known methods used in the art as liquid and powdered forms, It is preferable that the culture broth or concentrate of the strain JH7 of chromobacterium is adsorbed and dried on a carrier such as starch, crude protein or rock fractions. The carrier which can be used by mixing with the culture broth or concentrate of the strain of the present invention may be any carrier used in the art. Specific examples of the carrier that can be used in the present invention include seeds such as rice, wheat, corn, barley, soybeans, sorghum, millet, millet and the like, cereals such as cereals and potatoes, rootstocks such as sweet potatoes and cassava, (For example, powder), starches derived therefrom and derivatives thereof, and the like. In addition, diatomaceous substances such as agar, gelatin, pectate (polygalacturononate), chitosan, carboxymethylcellulose and derivatives thereof, clay minerals such as gelite, natural wax, natural gum, kaolin and bentonite, . These various carriers may be used individually, or a carrier of improved physical properties may be obtained by mixing two or more carriers in a suitable ratio. When such carriers are used, they can be metabolized by microorganisms and become a nutrient source. These carriers are highly viscous and thus increase the adhesiveness to the surface of plants.

The present invention can also include a dry product obtained by a drying method and a biological pesticide containing the strain or strain culture solution. The dried product can be prepared into a formulation selected from the group consisting of a wetting agent (WP), a solid phase (GM), a granular wettable agent (WG), a granule (GR), a powdered (DP) have. These bio-pesticide formulations are superior to conventional liquid formulations in terms of stability and physico-chemical properties and can be used for plant disease control.

A wetting agent in a biocidal pesticide refers to a formulation in which the pesticide is hydrated when diluted in water as a powder and the solid phase is a formulation in which a microorganism culture fluid is mixed with or adsorbed on a solid phase material and is not classified as a powder, granule or wettable powder. The granular wettable powders may be formulated in the form of a pesticide used as a granular phase by diluting with water, a granular form of the pesticide used as a granule used as a granule, a granule of a pesticide used as a granule, Means a formulation of pesticide used as a suspension by hydration before treatment with seed as a powder.

In order to control the plant disease using the microorganism preparation of the present invention, the microorganism preparation may be uniformly diluted with water and then sprayed onto crops, seeds of crops or cultivated land using an appropriate spraying device such as a power sprayer. When diluting the formulation of the present invention the water concentration, but can effective component adjusted to 10 ³ to 10 ⁵ cfu / ml, preferably 10 ⁴ cfu / ml and out to be a valid range biologically, but is not limited thereto.

The present invention also provides a method of controlling a plant disease comprising the step of treating an effective amount of the plant disease control composition to a plant part, soil or seed.

As a method for controlling the plant disease, a culture solution in which the strain of the present invention has been cultured, or a microorganism preparation using the strain may be immersed in the seeds of crops or crops, or may be carried out by spraying or spraying. In the case of the immersion method, the culture medium and the preparation can be poured into the soil around the plant or the seed can be immersed in the culture medium and the preparation. Plants that can be applied to the method of the present invention are not particularly limited.

In the method according to one embodiment of the present invention, the plant bottle may be, but is not limited to, ginseng roots rot caused by Shirindokapon death truck.

In addition, the present invention provides a method for producing a microbial preparation for controlling a plant disease comprising culturing the strain. Any of the methods known in the art can be used for the culture method of the above chromobacterium genus JH7 and the method for producing the microorganism preparation, and the method is not particularly limited.

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. Antibacterial activity of antagonistic bacteria against ginseng roots rot fungi

132 germs were isolated from ginseng cultivation soil in order to isolate germs with excellent antimicrobial activity against ginseng root rot disease. These bacteria were tested for the antimicrobial activity through confluent culture. The ginseng roots rot fungus was cultivated on a PDA (potato dextrose agar) medium at 21 ℃ for 10 days after using Cylindrocarpon destructans (KACC41077). A piece of cell with a diameter of 4 mm was placed on the center of a 9 cm PDA medium. Each bacterium was cultured in LB liquid medium at 28 ° C for 16 hours with shaking at 200 rpm. The prepared ginseng roots rot fungus was inoculated with PDE medium inoculated with 20 ㎕ of each bacterium by 4 mm holes at 3 cm intervals in the third row. After 15 days, mycelial growth length was measured. As a result, six out of 132 bacteria were cultured in confluent culture And mycelial growth was strongly inhibited. Among them, the JH7 strain showed a high mycelial growth inhibition ability of 50% or more (Table 1 and Fig. 1A). The JH7 strain exhibited a mucinous colony characteristic of a round yellow cream color in the LB solid medium (Fig. 1B).

^a Inhibition rate = [mycelial growth (control) - mycelial growth (experiment)] / mycelial growth (control) x 100

^b value is the mean ± standard deviation, which was calculated from three independent experiments. The other letters represent significant differences based on Duncan's multi-range test (p = 0.05).

Example 2. Identification of 16S rRNA Sequence Analysis of Selective Antagonistic Bacterium JH7

JH7 strain, which showed excellent antimicrobial activity through confluent culture, was selected for further biological control studies. Genomic DNA was extracted with Wizard genomic DNA purification kit (Promega, USA) for identification of JH7 strain. 1.4 kb of 16S rRNA gene was amplified by Thermal Cycler Thermal Controller 2720 (Applied Biosystems, USA) with universal primer 27F (5'-AGAGTTTGATCMTGGCTCAG-3 ': SEQ ID NO: 2) and 1492R (5'- TACGGYTACCTTGTTACGACTT- And sequenced to obtain a nucleotide sequence (SEQ ID NO: 1). The 16S rRNA sequence was subjected to BLAST searches at NCBI and EZbiocloud (https://www.ezbiocloud.net/). Each DNA sequence was aligned using ClustalW of MEGA 7, and the phylogenetic relationship was determined by the neighbor-joining method of 1,000 bootstrap. As a result, the JH7 strain was found to contain chromosomal bacterium KJ619639 (99%), C. violaceum ( HM449690, 99%), chromobacterium genus AB426118 (99% tikum was found (C. haemolyticum, KJ845679, 99% ), chromotherapy tumefaciens aqua tikum (C. aquaticum, JQ582944, 99% ) chromotherapy tumefaciens sp Im (E value of 0.0), including the (2 ).

Example 3. Physiological and biochemical analysis of the selected antagonistic bacterium JH7 using API 20NE

Biochemical and biochemical analyzes of JH7 strains identified as Chromobacterium sp. Strains were performed using API20NE (BioMérieux, Marcy l'Etoile, France) as a result of 16S rRNA sequencing. As a result, it has been found that d-glucose, 1-arabinose, d-mannitol, N-acetylglucosamine, potassium gluconate, capric acid, malic acid, trisodium citrate, Phenylacetic acid showed metabolic activity, while d-mannose, maltose, and adipic acid showed metabolic inactivation. In addition, although positive reactions were observed with potassium gluconate, d-glucose fermentation, arginine dihydrolase, urease and gelatin hydrolysis, tryptophan diamine, esculin hydrolysis esculin hydrolysis, and p-nitrophenyl-bD-galactopyranoside showed negative reaction. These results suggest that the differences in the five strains (urease, gelatin, 1-arabinose, d-mannitol and phenylacetic acid) and the strains of C. violaceum ( HM449690) And finally named as JH7 strain of chromobacterium.

Physiological and biochemical analysis of JH7 strain Substrate JH7 C. violaceum Potassium nitrate ^{+ a} + Tryptophan - - Glucose w + Arginine + + Urea + - Esculin - - Gelatin w - p-Nitrophenyl- [beta] -D-galactopyranoside - - Glucose + + Arabinose w - Mannose - - Mannitol + - N-acetyl-glucosamine + + Maltose - - Gluconate + + Caprate + + Adipate - - Malate + + Citrate + + Phenyl-acetate + - API 20 NE no. 5355557 5140555

^a +, full metabolism; -, not metabolized; w, weak metabolism.

Example 4. Analysis of antimicrobial activity of selected antagonistic bacterium JH7

JH7 strains (chitinase, protease, and cellulase). The chitinase activity was determined by colloid chitin agar medium supplemented with colloidal chitin (0.7 g / l K ₂ HPO ₄ , 0.3 g / l KH ₂ PO ₄ , 0.5 g / l MgSO ₄ .5H ₂ O, 0.01 g / l FeSO _{4 · 7H 2 O, 0.001 g} / l ZnSO 4, was used to make _{0.001 g / l MnCl 2, 15} g / l agar and 0.5 g of 1 dissolving the colloidal chitin l distilled water). The strain was inoculated with 20 ml of an antagonistic microorganism culture solution having an OD value of 1.0, and then cultured at 25 ° C for 3 days. The culture broth was corked with a cork borer. In order to examine cellulase activity, CMC agar medium was used in Sazci et al. (1986) (Sazci et al., 1986 Journal of Applied Bacteriology 61: 559-562) and inoculated in the same manner as above, And the cells were stained in a 0.1% Congo red solution for 15-20 minutes. Then, the cells were stained with 1 M NaCl solution for 15-20 minutes to decolorize the cells. In order to investigate the proteolytic activity, LB medium supplemented with 3% skim milk powder was inoculated with the bacterium in the same manner as described above. After culturing for 3 days, clearance zone formation was observed to examine the protease activity. A simplified modified CAS (Chrome Azurol Sulfonate) assay was used to study Siderophore activity. CAS (Sigma, USA) dye solution was prepared by dissolving 60.5 mg of CAS in 50 ml of distilled water and dissolving 72.9 mg of HDTMA in 40 ml of distilled water to prevent light from entering. Then, 35 g of LB agar was dissolved in 900 ml of distilled water, and the three solutions were mixed and autoclaved. After that, 1 mM FeCl ₃ .6H ₂ O was dissolved in 10 ml of HCl solution (10 mM) to adjust the pH to 6.8 and autoclaved separately. The two sterilized high pressure sterilized solutions were cooled to 50 ° C, mixed slowly to prevent foaming, and placed in Petri dishes to make CAS flat plate culture medium. Thereafter, the bacteria were inoculated in the above manner and cultured at 25 ° C for 5 days. Then, the presence or absence of clear zone formation was determined to determine the production of siderophores. In order to investigate whether antagonistic microorganisms can degrade weakly soluble phosphate, activity assays were carried out using Pikovskaya's agar medium containing insoluble phosphoric acid. This medium was prepared by the method of Gaind and Gaur (1991) (Gaind & Gaur 1991 Plant Soil 133: 141-149) and inoculated with the bacterial culture solution as described above. After incubation at 25 ° C for 15 days, the activity of phosphoric acid-solubilization was determined with or without clear zone formation.

As a result, JH7 decomposed chitin but did not decompose cellulose. And strong protein resolution. It was found that the halo region was produced from the CAS medium to produce sea sylophor. In addition, the ability to decompose insoluble phosphoric acid was also observed.

Example 5: Production of spores and inhibition of spore formation and spore formation by selective antagonistic bacterium JH7 treatment

Spores play an important role in causing disease in most plant pathogenic fungi. JH7 was treated to determine the effect on spore production (Fig. 4A). As a result, spores of 1.7, 3.7, and 2.7 × 10 ⁴ / ml were present at 3, 6, and 9 days after treatment at 1% concentration, respectively. The inhibition rate of spore formation was 97.4%, 97.6% and 98.7%, respectively. In the 0.1% and 0.01% treatment groups, the inhibition rates were 76.4% and 71.2% at 3 days, respectively, and 82.9% and 87.8% at 6 days and 9 days, respectively. As a result, when JH7 was treated at a concentration of 1%, spore formation was almost inhibited, and a suppression rate of spore formation of 80% or more was obtained at a concentration of less than 0.1%.

The spore germination was tested at the early stage of the onset to determine whether treatment with JH7 could inhibit spore germination (FIG. 4B). In the control, the spores of ginseng root rot fungus were germinated at 94% for 2 hours. However, in the JH7 treated with O.D value of 1.0, only 1% germinated for 2 hours and showed 98.9% germination inhibition rate. O.D 0.1 and 0.01, respectively, showed 80.9% and 41.5% germination inhibition rate in JH7 treated plants, respectively. As a result, it was found that treatment of JH7 with O.D 1.0 can suppress spore germination almost, and that treatment with less than 0.1 slows or suppresses germination.

Thick spore was tested as a primary source of ginseng root rot fungus, inhibiting formation of thick spores by treatment with JH7 (FIG. 4C). When JH7 was treated at a concentration of 1%, spore formation was inhibited, so that it was inoculated at a concentration of 1% to observe whether thick membrane spores were formed. As a result, it was observed that thick membrane spore was formed in the untreated zone, but JH7 was also shown to suppress the formation of thick membrane spore because no thick membrane spore was observed in JH7 treatment.

Example 6. Antimicrobial activity spectrum of the selective antagonistic bacterium JH7

Studies on the antimicrobial activity of JH7 strain, which has excellent antimicrobial activity against ginseng root rot fungus, against other phytopathogenic fungi were carried out. As a result, the JH7 strain showed antibacterial activity against all 10 test pathogens. Among them climb belonging to Dottie for Oh My three tests (Dothideomycetes) also sports Leeum Cuckoo Mary num strongest antibacterial in (Cladosporium cucumerinum), x vertical hilrum Tour sikum (Exserohilum turcicum), nose Clio bolus hetero Our Perth (Cochliobolus heterostrophus) Lt; / RTI > Rhizoctonia solani , Botrytis cinerea and Magnaporthe oryzae showed moderate antimicrobial activity and choletto tree belonging to Sordariomycetes , Colletotrichum acutatum , Fusarium oxysporum and Fusarium solani showed weak antimicrobial activity.

Taxonomy JH7 Class Genus, Species Dothideomycetes Cladosporium cucumerinum ^{++++ a} " Exserohilum turcicum ++++ " Cochliobolus heterostrophus ++++ Agaricomycetes Rhizoctonia solani +++ Leotiomycetes Botrytis cinerea +++ " Sclerotinia sclerotiorum ++ Sordariomycetes Magnaporthe oryzae +++ " Colletotrichum acutatum ++ " Fusarium oxysporum ++ " Fusarium solani +

^a Determine the average diameter of the inhibition clear zone and determine +, <5 mm; ++, 6-10 mm; +++, 11-15 mm; ++++, > 15 mm.

National Center for Agricultural Genetic Resources KACC92211P 20171116

<110> KNU-Industry Cooperation Foundation <120> Chromobacterium sp. JH7 strain having antifungal activity against          plant pathogens and uses thereof <130> PN17510 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 1342 <212> DNA <213> Chromobacterium sp. <400> 1 ctcagattga acgctggcgg catgctttac acatgcaagt cgaacggtaa cagggtgctt 60 gcaccgctga cgagtggcga acgggtgagt aatgcatcgg aatgtaccgt gtaatggggg 120 atagctcggc gaaagccgga ttaataccgc atacgccctg agggggaaag tgggggaccg 180 taaggcctca cgttatacga gcagccgatg tctgattagc tagttggtgg ggtaaaggct 240 caccaaggct tcgatcagta gcgggtctga gaggatgatc cgccacactg ggactgagac 300 acggcccaga ctcctacggg aggcagcagt ggggaatttt ggacaatggg cgcaagcctg 360 atccagccat gccgcgtgtc tgaagaaggc cttcgggttg taaaggactt ttgtccggga 420 gcaaatccca gtggttaata cctactgggg ctgagagtac cggaagaata agcaccggct 480 aactacgtgc cagcagccgc ggtaatacgt agggtgcaag cgttaatcgg aattactggg 540 cgtaaagcgt gcgcaggcgg ttgtgcaagt ctgatgtgaa agccccgggc ttaacctggg 600 aacggcattg gagactgcat gactagagtg cgtcagaggg gggtagaatt ccgcgtgtag 660 cgtgaaatg cgtagagatg cggaggaata ccgatggcga aggcagcccc ctgggatgac 720 actgacgctc atgcacgaaa gcgtggggag caaacaggat tagataccct ggtagtccac 780 gccctaaacg atgtcaacta gctgttgggg gtttgaatcc ttggtagcga agctaacgcg 840 agaagttgac cgcctgggga gtacggccgc aaggttaaaa ctcaaaggaa ttgacgggga 900 cccgcacaag cggtggatga tgtggattaa ttcgatgcaa cgcgaaaaac cttacctggt 960 cttgacatgt aacgaacgcc gcagagatgt ggtggtgccc gaaagggagc gttaacacag 1020 gtgctgcatg gctgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080 gcaacccttg ccattagttg ccatcattaa gttgggcact ctaatgggac tgccggtgac 1140 aaaccggagg aaggtgggga tgacgtcaag tcctcatggc ccttatgacc agggcttcac 1200 acgtcataca atggtcggta cagagggtcg cgaagccgcg aggtggagcc aatctcataa 1260 aaccgatcgt agtccggatc gcactctgca actcgagtgc gtgaagtcgg aatcgctagt 1320 aatcgcagat cagcatgctg cg 1342 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 agagtttgat cmtggctcag 20 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 tacggytacc ttgttacgac tt 22

Claims (10)

A Chromobacterium sp. JH7 strain having an antifungal activity against a plant pathogenic agent and having a deposit number of KACC92211P. delete The strain according to claim 1, wherein the fungus is Dothideomycetes , Agaricomycetes , Leotiomycetes and Sordariomycetes . . The method according to claim 1, wherein the fungi are selected from the group consisting of Cylindrocarpon destructans , Cladosporium cucumerinum , Exserohilum turcicum , Cochliobolus heterostrophus , Rhizoctonia solani , Botrytis cinerea , Sclerotinia sclerotiorum , Magnaporthe oryzae , Choletotrichum , Aqua tatum (Colletotrichum acutatum), Fusarium oxy sports rooms (Fusarium oxysporum and Fusarium solani . The strain according to claim 4, wherein the strain has chitinase and protease releasing ability, sea flavor producing ability, insoluble phosphate decomposing ability, spore formation inhibiting ability, spore germination inhibiting ability and chlamydospore generation inhibiting ability. A plant disease controlling composition comprising the strain according to any one of claims 1 to 7, a culture solution thereof or a culture medium extract as an active ingredient. The plant according to claim 6, wherein the plant is selected from the group consisting of Cylindrocarpon destructans , Cladosporium cucumerinum , Exserohilum turcicum ), Cochliobolus heterostrophus heterostrophus , Rhizoctonia solani , Botrytis &lt; RTI ID = 0.0 &gt; cinerea ), Sclerotinia sclerotiorum , Magnaporthe oryzae), Collet sat tree glutamicum Aqua tatum (Colletotrichum acutatum), Fusarium oxy sports rooms (Fusarium wherein the plant disease is caused by a plant disease caused by Fusarium solani or Fusarium solani . [Claim 7] The composition according to claim 6, wherein the plant disease is ginseng root rot disease. A method for controlling a plant disease comprising the step of treating an effective amount of the plant disease control composition of claim 6 to a plant part, soil or seed. 10. The method according to claim 9, wherein the plant disease is ginseng root rot disease.

Images