KR100760526B1 - 119 119 fusarium oxysporum ef119 strain producing bikaverin microorganism formulation for controlling plant diseases containing bikaverin or ef119 strain and method for controlling plant diseases using same - Google Patents

Abstract

A novel Fusarium oxysporum EF119 strain and bikaverin produced by the same are provided to control the plant diseases environment-friendly by inhibiting the growth of various plant pathogens in vivo and in vitro, thereby capable of producing organic agricultural products. A non-pathogenic Fusarium oxysporum EF119 strain producing bikaverin is deposited as a deposition number of KCTC 10926BP. The microorganism preparation for controlling plant diseases such as a plant disease caused by Oomycetes, Colletotrichum crassipes, anthracnose, Plasmodiophora brassicae, and Streptomyces scabies comprises the Fusarium oxysporum EF119 strain, a culture solution thereof, a hypha thereof or a pore thereof as an effective ingredient. To control the plant diseases, a plant is treated with the microorganism preparation.

Description

Microbial preparation for plant disease control, including Fuzarium oxysporum EF119 strain, vicarin or EF119 strain, which produces vicaberine, and method for controlling plant disease using the same DISEASES CONTAINING BIKAVERIN OR EF119 STRAIN AND METHOD FOR CONTROLLING PLANT DISEASES USING SAME}

Figure 1 is a photograph of the Fusarium oxysporum EF119 strain growing on potato dextrose agar medium.

Figure 2 compares the nucleotide sequences of the Fussium oxysporum EF119 strain and the Fussium oxysporum Fo47 strain.

Figure 3 is a photograph of the EF119 strain inhibits mycelial growth of cucumber mozzarella bacteria.

Figure 4 is a photograph showing the control effect of bicaberin tomato late blight. The plant on the left is treated with the control chlorothalonil (50 μg / ml), and the middle three plants are treated with vicaberine at levels 33.3, 100 and 300 μg / ml from the left. Plants are not treated with any drugs.

The present invention provides a microbial agent comprising a non-pathogenic Fusarium oxysporum EF119 strain, a non-caberine or an EF119 strain, which produces the anti-cababerine, and a method for biologically controlling plant diseases using the microbial agent. It is about.

Pesticide use is inevitable to increase yields in crop cultivation, but misuse of synthetic pesticides, which make up the majority of pesticides, raises various problems such as soil, water and agricultural pollution, toxicity, disturbance of ecosystems, and emergence of resistant strains. . In recent years, research on the development of biopesticides using antagonistic microorganisms, which is one of the methods for solving these problems, has been actively conducted worldwide.

On the other hand, Huzirum oxysporum strains are known as phytopathogens that cause vascular wilt, a soil infectious disease in various plants. Recently, however, it has been applied as a biopesticide, in particular the Fuzium oxysporum Fo47 strain has been developed as a microbial fungicide and is currently present in the Fussium oxysporum , Fusarium moniliforme , and Gibberella fujikuroi. It is used to control the wilting disease caused by (Copping, LG (ed), The BioPesticide Manual, 3rd Edition (2004)). Three mechanisms for controlling wilted ill disease by non-pathogenic fusidium oxysporum have been reported: competition with pathogens in the roots, competition with pathogens in the root surface, and resistance to induction to plants. It has also been reported that the non-pathogenic fusidium oxysporum Fo47 strain also controls cucumber mozzarlock disease caused by Pythium ultimum strains (Benhamou, N. et al. , Appl. Environ.Microbiol . 68: 4044-4060). (2002)). Activity against this plant disease is reported to be due to a combination of antimicrobial activity, parasitic and induced resistance of non-pathogenic strains against pathogens. Benhamou et al. Have reported on the action of antimicrobial agents on Pisium Ultimum bacillus, but it is not known which ones.

Bicaberine is described in Cornforth, JW et al. , J. Chem. Soc. 2786-2788 (1971); de Boer, JJ et al. , J. Chem. Soc. 2788-2791 (1971); Brewer, D. Et al., J. Antibiotic. 26: 778-781 (1973); and Bell, AA et al . , Pest. Manag. Sci. 590: 736-747 (2003)) and Gibberella Fujikuroi (Balan, J. et al., Folia) Microbiol. 15: 479-484 (1969); Kjaer, D. et al. , J. Chem. Soc. 2792-2797 (1971); Giordano, W. et al. , FEMS Microbiol. Lett. 173: 389-393 (1999); And Escamilla-Silva, E. et al., World J. Microbiol. Biotechnol. 17: 469-474 (2001)). To date, vicarberin has shown strong activity against Leishmania brasiliensis protists (Balan, J. et al . , Folia Microbiol. 15: 479-484 (1969)), and in vitro against various cancer cells. It is known to be toxic (Fuska, J. et al., Neoplasma 22: 335-338 (1975)). Bicaberin is also known to promote intracellular vesicles, a symptom of aging, in the mycelial tip of Aspergillus niger (Cornforth, JW et al. , J. Chem. Sco. 2786-2788 (1971). )).

Among the endophytes isolated from various plants, the team selected fungi that showed excellent antagonism against various plant diseases including tomato late blight. It has been found to show high activity against cucumber anthrax and Chinese cabbage root gall disease. In addition, the antimicrobial substance produced by this strain is bicaberine, which inhibits the growth of various plant pathogens in vitro, and invented the present invention by inventing for the first time the high activity against tomato late blight in vivo. Completed.

It is an object of the present invention to provide a novel Fuzium oxysporum strain that produces vicaberine.

Another object of the present invention to provide a microbial agent for controlling plant diseases comprising the strain or vicarberine.

Still another object of the present invention is to provide a method for controlling plant diseases using the microbial agent.

In accordance with the above object, the present invention provides a viumum oxysporum EF119 strain (KCTC 10926BP) after producing the bicarberine of formula (I):

In order to achieve the above another object, the present invention provides a microbial agent for controlling plant diseases containing the strain or a non-caberine derived therefrom as an active ingredient.

In order to achieve the above another object, the present invention provides a method for controlling plant diseases using the microbial agent.

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

Huzium oxysporum EF119 strain of the present invention has the following morphological and biochemical characteristics.

Fuzium oxysporum EF119 strain produces a lot of white aerial hyphae on potato dextrose agar (PDA) medium, and the color of the back side of the medium becomes purple over time (FIG. 1). In addition, long-term incubation in potato dextrose agar medium produces microconidia, macroroconidia and chlamydospore. The Fussium oxysporum EF119 strain was identified with GeneBank's Fusarium oxysporum f. Sponi melonis in the nucleotide sequence (SEQ ID NO: 1) of ITS1- 5.8S-ITS2 . Sequence homology of% is shown.

The strains identified in the present invention were isolated from the red pepper roots and synthesized the morphological characteristics and ITS1-5.8S-ITS2 sequence data of the strains, and were determined to be Fussium oxysporum. It was named Room EF119 and deposited on March 22, 2006 as a deposit No. KCTC 10926BP with the Korea Biotechnology Research Institute Gene Bank.

Fuzilium oxysporum EF119 strains include tomato phytophthora infestans , pepper phytophthora capsici , cucumber moss ultimum , cucumber fungi ( Pseudoperonospora cubensis ), and grape vine fungi ( Plasmopara viticola ). As well as plant diseases caused by fungal fungi, cucumbers such as Colletotrichum lagenarium , red pepper anthrax ( C. gloeosporioides, C. acutatum ), Chinese cabbage root ( Plasmodiophora brassicae ) and potato flour ( Spogospora subterranea ) To control plant diseases. In addition to the above pathogens, vicarberin isolated from the Fussium oxysporum EF119 strain is a variety of germs, such as Magnaporthe grisea , Alternaria mali , and Tomato Blight fungus ( Botrytis cinerea ). Control plant diseases by

Huzurium oxysporum EF119 strain of the present invention is a microorganism preparation for plant disease control by formulating it as a powder, pellets, granules or solution by mixing the carrier itself, the culture solution, or extract of the strain itself, or mycelia and spores alone It can be used as, the carrier may be used, such as water, white carbon, kaolin. In addition, it is also possible to prepare a microbial preparation for controlling plant diseases by formulating only the vicarberin as described above.

Formulated microbial agents, on the other hand, can be treated on the soil in which the plant is growing or on the surface of the growing plant to control the growth of the plant caused by plant diseases and thereby kill the dead.

Huzalium oxysporum EF119 strain of the present invention can be treated to the target plant as a formulation of 1 × 10 ³ propagules / ml to 1 × 10 ⁸ propagules / ml concentration. In addition, vicarinine can be treated to plants as a preparation at a concentration of 10 μg / ml to 500 μg / ml.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1: Isolation of Huzium oxysporum EF119 Strain

In May 2001, three healthy pepper plants per package were taken from one pepper sample from a number of pepper packages grown in farms around Chungcheongbuk-do. After taking samples from a total of eight packages, each sample was divided into three parts, that is, leaves, stems, and roots, and then two surface sterilizations were performed on 24 samples to separate endophytic fungi. The law was enforced. The leaves were soaked in 2% sodium hypochlorite (NaOCl) for 10 seconds, washed twice with sterile water, and then crushed into small pieces. Corn flour-malt extract-yeast extract agar added with 50 ㎍ / ml chloramphenicol (Sigma, USA) The medium (CMA medium: cornmeal malt extract agar medium: 17.0 g corn flour, 20.0 g malt extract, 2.0 g yeast extract, 1.0 liter distilled water) was dentified. Stem and root tissues were briefly immersed in ethanol and then flame sterilized and then wound on CMA medium to which chloramphenicol (50 ㎍ / ml) was added. The bacteria were isolated while incubated under 25 ° C constant temperature. The isolates were again subjected to single mycelial separation on PDA medium. A total of 89 fungi were isolated, and for the long term storage of the strain, the tip of the strain cultured in PDA medium for 7 days was removed with a cork borer (8 mm in diameter) and sterilized distilled water and 6% dimethylsulfoxide (DMSO). The solution was added and stored at room temperature and -70 ° C, respectively.

Example 2: Incubation and in vivo of isolated strains in vivo Antimicrobial Activity

The control activities of four plant diseases including tomato late blight, cucumber anthrax, tomato ash fungus and barley powdery mildew were investigated as follows.

Sterilized 200 ml potato dextrose broth (PDB) was inoculated with four pieces of the cork-cutter off the tip of the strain growing on the PDA medium and incubated for 10 days at 25 ℃, 150 rpm. The culture solution was centrifuged at 10,000 × g for 15 minutes, and only the supernatant was taken as a liquid culture sample.

In order to select strains with antimicrobial activity, in vivo antimicrobial activity assays of liquid culture samples of 89 endogenous fungi were carried out on the four plant diseases. After liquid culture was ground with a homogenizer, Tween 20 was added at a concentration of 250 μg / ml to 30 ml of this culture stock solution or its 3-fold or 9-fold dilution, followed by 24 hours prior to inoculation of pathogens. It was. Cucumis ( Cucumis sativus varieties), Tomato (Lycopersicon esculentum varieties: Western light) and Barley ( Hordeum sativum varieties: Eastern barley) seeds are sown in 4.5 cm diameter plastic pots containing about 70% of horticultural soil and grown in greenhouses (25 ± 5 ° C.) for 1 to 3 weeks.

Cucumber anthrax was sprayed with conidia spore suspension (10 ⁶ spores / ml) of cucumber anthrax ( colletotrichum obiculare ; obtained by Korea Research Institute of Chemical Technology) on three-leaf cucumber seedlings, and then kept in a 25 ℃ constant temperature chamber for 2 days in a 25 ℃ constant temperature and humidity chamber. Induced onset for 3 days. The tomato late blight was inoculated on a two-layered tomato seedling in a 20 ° C wet room after spraying with a distilled suspension suspension extracted from the rotiferous sap suspension (3 × 10 ⁴ sagittal sac / ml) of Phytophthora infestans (obtained at Gangneung University). Induced. On the other hand, tomato gray mold disease was developed in a wet room after treatment with spore suspension (5 × 10 ⁵ spores / ml) of two-stage tomato seedlings treated with Botrytis cinerea (obtained from Korea Research Institute of Chemical Technology). Barley powdery mildew was inoculated with the spores of the barley powdery mildew ( Blumeria graminis f. Sp. Hordei ; source: Korea Research Institute of Chemical Technology) passaged in host plants at the first stage of barley seedlings. As a control, 30 ml of Tween 20 solution (250 μg / ml) added with 1% DMSO was used. Cucumber anthrax was investigated 5 days after inoculation, tomato gray mold and tomato late blight 3 days after inoculation, and barley powdery disease after 7 days after inoculation.

As a result of screening, the most highly active strains for tomato late blight and cucumber anthrax were separated and named as EF119, and their control values (%) are shown in Table 1 below.

Example 3: Identification of EF119 Strain

(1) Culture and Morphological Characteristics

The EF119 strain cultured in PDA medium had a white mycelium color as shown in FIG. 1 and the color of the back side of the medium changed to purple over time. After incubation time, microconidia, macroconidia and chlamydospore were produced on the cells.

(2) ITS1-5.8S-ITS2 sequencing

For accurate identification of the strain, EF119 strain was inoculated on a cellophane-plated PDA medium and incubated at 25 ° C. for 4 days, and DNA was extracted from the mycelium using AccuPrep Genomic DNA Extraction Kit (obtained by Bioneer Korea). From the extracted DNA, ITS4 and 5 primers (White, TJ et al., PCR protocols: a guide to methods and application , pp.315-322 (1990)) were used to prepare ITS1-5.8S-ITS2 of nuclear ribosomal DNA (nuclear rDNA). After amplification, the purity was checked, and then the sequence was determined using an ABI3700 automated DNA sequencer (obtained from Applied Biosystems, USA). Nucleotide sequence analysis of ITS1-5.8S-ITS2 showed 99% sequence homology with Geneusa's Fusarium oxysporum f. Sp. Melonis AY188919 strain . Was named Huzium oxysporum EF119. The ITS1-5.8S-ITS2 nucleotide sequences (SEQ ID NOs: 1 and 2) of the Furizium oxysporum EF119 strain and the Furizium oxysporum Fo47 strain, which are used as non-pathogenic strains, are shown in FIG. 2.

According to the above results, the EF119 strain was identified as Fuzarium oxysporum, and it was deposited with KCTC 10926BP as of March 22, 2006 to the Korea Biotechnology Research Institute Gene Bank.

Example 4 In Vitro of the E.F. in vitro Antibacterial activity

To investigate the in vitro antimicrobial activity of E. coli strains against fungal mycobacteria, tomato phytophthora infestans (Gangneung University), phytophthora capsici (Chungnam National University) and Pythium ultimum ; : Korea Research Institute of Chemical Technology) was incubated with EF119 strain in agar medium. That is, inoculated with agar plugs (agar plug) of two strains at intervals of about 4 cm in agar medium plate 9 cm in diameter and incubated. Tomato blight was carried out on V-8 juice agar medium (V-8 juice 200 ml, CaCO ₃ 4.6 g, agar 20 g, distilled water 800 ml), and pepper blight and cucumber mozolox bacterium were carried out on PDA medium. It was.

As a result, the EF119 strain showed antagonistic activity in tomato agar, pepper blight and cucumber mozolox bacteria in agar medium. Figure 3 shows the mycelial growth of cucumber mozzarella bacteria inhibited by the EF119 strain.

Example 5: In Vivo Antimicrobial Activity of Chinese Cabbage Root-knock Disease and Red Pepper Plague Disease of EF119 Culture

To test the effect of EF119 culture on Chinese cabbage root gall, cabbages were sown in horticultural soil and grown for 2 weeks before transplanting into plastic pots (7.0 cm in diameter) containing field soil inoculated with cabbage root gall bacteria. Chinese cabbage root inoculation was inoculated by mixing 100 g of the diseased tissue collected from the package in 10 liters of field soil and mixing well. After transplanting, 30 ml of the solution in which Tween 20 was added at a level of 250 µg / ml was added to the culture stock prepared as in Example 2, and then irrigated in a pot. Untreated groups were treated with tween 20 million at the 250 μg / ml level. Treated plants were grown for 4 weeks in a constant temperature and humidity room (20 ° C), and the incidence in roots was measured.

As a result, the culture solution of EF119 showed a control effect of 50%.

On the other hand, in order to test the effect of EF119 culture on the pepper blight, the culture medium containing 250 ㎍ / ㎖ of Tween 20 at 10 ml per pot in the pepper plant of the primary basin grown in a plastic pot with a diameter of 7.0 cm Stock solutions and 10-fold dilutions were irrigated. One day later, phytopathogens were inoculated by irrigation of 1 × 10 ⁴ sacscapillary suspension (1 × 10 ⁴ sacs / ml) in each port by irrigation. After the inoculation, the incidence was evaluated after 8 days with the bottom irrigation in the greenhouse.

As a result, the culture stock and 10-fold dilution showed 56% and 22% control effects, respectively.

Example 6: Antibacterial Activity of Mycelia and Culture Filtrate of EF119 Strain

In order to investigate whether the antimicrobial material is present in the culture medium or the culture filtrate in EF119 strain, the tip of the mycelium growing in PDA medium in 200 ml PDB medium in 500 ml Erlenmeyer flask was removed with a cork cutter. After inoculating the four pieces were shaken and incubated for 14 days at 25 ℃, 150 rpm. After incubation, 30 ml of the culture solution was taken, and divided into mycelia and culture filtrate using filter paper. Mycelia were diluted again with 30 ml of distilled water. The culture, filtrate and mycelium samples were diluted 10-fold and 50-fold with distilled water, and then Tween 20 was added at a level of 250 ㎍ / ml, respectively, and the control effect against tomato late blight was investigated in the same manner as in Example 2. Is shown in Table 2 below.

As a result of the above experiment it can be seen that the antimicrobial material is present in both the culture filtrate and mycelium.

Example 7: Isolation, Purification and Structure Determination of Antimicrobials

Four pieces of the tip of the strain growing in the PDA medium with a cork cutter were inoculated into 200 mL of PDB medium, followed by incubation at 25 ° C. for 150 days at 150 rpm. Cultures were taken and inoculated in 3 L PDB medium in 5 L jar fermentor at a concentration of 1% (v / v). After inoculation, the cells were shaken at 25 ° C. for 5 days at 150 rpm. After filtration of the culture, the filtrate was partitioned three times with the same amount of chloroform, and the organic solvent layer was concentrated under reduced pressure. The concentrated sample was redissolved again with a small amount of chloroform, and then a small amount of diethyl ether was added to precipitate. The precipitate was recrystallized with chloroform again to obtain about 250 mg of pure material. To investigate the purity of this material, chloroform-acetone-formic acid (84: 15: 1, v / v / v) was used as a developing solvent and TLC (20 × 20 cm, thickness 0.25 mm, Kiesel gel 60 F254; Merck analysis showed a single substance.

Purely separated and purified one active material was analyzed by mass spectrometry (JEOL JMS-DX303; obtained by Nippon Electronics Co., Ltd.). As a result, the molecular weight was 382 Daltons and the mass spectrum was found to be the same as bicaberin by a Wiley database. Also, to confirm the structure, 5 mg of the material was dissolved in 0.5 ml of chloroform, placed in a 5 mm NMR tube, and subjected to NMR analysis [Bruker-AMX 500 (500 MHz)], and ¹ H-NMR was measured at 500 MHz. As a result, one methyl group (3H, 2.86, s), two methoxy groups [3H × 2, 3.93 (s), 3.95 (s)] and three aromatic rings of hydrogen [1H × 3, 6.77 (s), 6.92 (s), 6.35 (s)] peaks were consistent with the data of Bicaverin, as revealed by Kjaer et al. ( J. Chem. Soc. , 2792-2797 (1971)).

Example 8 In Vivo Antimicrobial Activity of Bicaberin

In order to investigate the control activities of various plant diseases of vicarberin isolated from the Fuzium oxysporum EF119 strain, experiments were conducted on plant diseases such as rice blast, rice leaf blight and tomato late blight. First, bicaberine was dissolved in acetone and diluted with 250 μg / ml Tween 20 solution to prepare final concentrations of 300 μg / ml, 100 μg / ml and 33.3 μg / ml. At this time, the final concentration of acetone was to be 50% level. At this time, a solution in which 15 ml of acetone (final concentration 50%) and 7.5 mg of Tween 20 (final concentration 250 µg / ml) was added to 15 ml of distilled water was used as a negative control. The non-caberine samples and the control samples thus prepared were sprayed onto the leaf surface of the plant seedlings one day before the inoculation of the pathogen, and then dried at room temperature for 24 hours. Activity test for tomato late blight was carried out as in Example 2.

The rice used in the experiment for rice blast and rice leaf blight ( Oryza sativa varieties: Nakdong) is filled with a soil pot or horticultural soil in a plastic pot of 4.5 cm in diameter about 70%, and then sowing seeds and greenhouses at 25 ± 5 ° C. Cultivation for 1-4 weeks. Rice blasts are sprayed with 3-4 leaf seedlings and sprayed with a spore suspension (5 × 10 ⁵ spores / ml) of Magnaporthe grisea (Korea Research Institute of Chemical Research), which is the causative agent of blasting disease, After the wet treatment, the incubation was induced by incubation for 5 days in a constant temperature room of 25 ℃. Rice leaf blight is a cultured product obtained by culturing 7 days of cultivation of Thanatephorus cucumeris , a causative agent of leaf blight, in medium (90 g of wheat bran, 15 g of chaff, 100 ml of distilled water). The seedlings were inoculated to 5-leaf seedlings and wet-treated for 4 days on 25 ° C., and then incubated in a constant temperature room at 25 ° C. for 4 days to induce the onset.

As a control drug, chlorothalonil, a representative agent of tomato late blight, was used, and the area ratio of rice blast was 5 days after inoculation, 7 days after inoculation of rice leaf blight, and 3 days after inoculation of tomato late blight. From the control value (%) to calculate the results are shown in Table 3 below.

As shown in Table 3 above, vicarberin showed a particularly high activity against tomato late blight.

Example 9 In Vitro Antimicrobial Activity of Bicaberin

Magnaporthe grisea (obtained from: Korea Research Institute of Chemical Research) on a solid medium containing non-caberine in order to measure the in vitro antimicrobial activity against the non-caberine, which has been found to have an excellent effect against tomato late blight in the in vivo experiment. , Phytophthora capsici (obtained from Chungnam National University), Colletotrichum gloeosporioides (obtained from Korea Research Institute of Chemical Technology), tomato tolerant bacterium ( Phytophthora infestans ; obtained from Gangneung National University), apple tree spotted deciduous bacterium ( Mycelia for seven plant pathogens, including Alternaria mali , Kyung-Nong Co., Ltd., Botrytis cinerea ; KIM, Pythium ultimum , and Korea Research Institute of Chemical Research. Growth inhibition effect was investigated. Tomato blight was used for V-8 juice agar, and the other six phytopathogens were potato agar. After sterilization of each medium, bicaberine dissolved in dioxane (dioxane) was added to the medium to solidify to 9 μg / ml before solidification. 1% of dioxane was added to the untreated section. After killing the mycelia of each bacteria in the middle of the medium, tomato late blight and tomato gray fungus were incubated at 20 ℃, the remaining bacteria were incubated at 25 ℃. After incubating for 2 to 6 days, the mycelial growth was measured, and the mycelial growth inhibition rate was examined based on the degree of mycelial growth in the non-treated group, and the results are shown in Table 4 below.

As shown in Table 4, vicarberine showed mycelial growth inhibitory activity against various phytopathogens, and in particular, it can be seen that the inhibitory activity against pepper anthrax was excellent.

On the other hand, the effect of vicarinine tomato late blight on the germination of juvenile sac was investigated. The spermatozoon was harvested using sterile water from the culture of the pathogen cultured in oatmeal agar medium for 12 days, and then filtered with gauze, and then the concentration was adjusted to 10 ⁴ spermatozoa / ml. 10 µl of the vicarinine solution was added to 990 µl of the capsular bag suspension, and then 50 µl was added to the hole-slide glass. Thus, the dicarbane dissolved in dioxane was treated to final concentrations of 9, 3, and 1 μg / ml (final concentration of dioxane = 1%), and in the control group, only 1% of dioxane was treated. The treated hole-slide glass was placed in a sealed container, in which a paper containing water was placed on the bottom to maintain 100% humidity. After incubating the sealed vessel for 6 hours at 20 ° C., germination of the capsular bag was observed under an optical microscope. The treatment was repeated three times for each treatment, and 100 drip sacs were observed for each treatment. The germination rate of the zygote capsule germination rate in the non-caberine experimental group was compared with that of the control group, and the germination inhibition rate (%) was calculated, and the results are shown in Table 5 below.

As shown in Table 5 above, vicarinin inhibited germination of tomato plaques by about 37% even at very low concentrations of 1 μg / ml.

Example 10 Pathogenicity Study of Various Plants of EF119 Strain

To investigate the pathogenicity of the EF119 strain, tomatoes ( Lycopersicon esculantum varieties: Seogwang), red peppers ( Capsicum annum varieties: Bugang), Chinese cabbage ( Brassica campestris varieties: Sambokgut), cucumbers ( Cucumis sativus varieties: whitening tea), radish ( Raphanus) Pathogenicity experiments were carried out on eight plants, including sativus varieties: Heil radish), watermelon ( Citrullus vulgaris varieties: beetroot ), beans ( Glycine max varieties: golden soybean), and sesame seeds ( Sesamum indicum varieties: remaining). Each plant was sown in horticultural soil and grown in a greenhouse until one leaf of the main leaf emerged. When the first leaf came out, the roots were washed with tap water, and the ends of the roots were cut and cut out. After culturing the EF119 strain for 7 days in PDA medium, spores were harvested and inoculated by immersing the wounded plant roots for about 10 minutes in a spore solution suspended in distilled water at a concentration of 10 ⁶ spores / ml. Inoculated seedlings were transplanted into pots containing fresh soil (7.0 cm in diameter) and grown for 28 days and examined for incidence every 7 days.

As a result, the EF119 strain was found to be no pathogenic to the tested plants.

As described above, the Huzurium oxysporum EF119 strain of the present invention and the vicarinine produced by the strain inhibit the growth of various plant pathogens in vitro and in vivo, and thus use the culture medium, mycelium or spores of the strain, or By using environmentally friendly plant disease control can produce high value-added organic products.

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Claims (9)

Nonpathogenic Huzium oxysporum EF119 strain (KCTC 10926BP) that produces vicaberin. Huzurium oxysporum EF119 strain (KCTC 10926BP), or its culture, mycelium or spores as an active ingredient, a plant disease control agent. The method of claim 2, Plant disease, characterized in that the plant disease caused by Oomycetes (cucumber anthrax, pepper anthrax, cabbage root gall disease or potato flour beetle disease). The method of claim 2, Huzurium oxysporum EF119 strain comprising a concentration of 1 × 10 ³ propagules / ml to 1 × 10 ⁸ propagules / ml. delete delete The method of claim 3, The plant disease caused by the bacterium fungi is tomato late blight, pepper late blight, cucumber mozalok disease, cucumber downy mildew or vine downy mildew. delete Huzurium oxysporum EF119 strain (KCTC 10926BP), its culture, mycelium or spores to the plant, characterized in that the plant treatment method.

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