KR101287141B1 - Streptomyces griseus BIG105 for controlling plant diseases and uses thereof - Google Patents

Abstract

The present invention is Ralstonia Solanacea Rum solanacearum ), Phytophthora capsici ), Sclerotium ( Sclerotium) cepivorum , Rhizoctonia solani ), Fusarium oxysporum or Sclerotinia sclerotinia sclerotiorum) Streptomyces draw with the control effect against a three-house (Streptomyces griseus ) It relates to a BIG105 strain, a plant disease control composition comprising the strain as an active ingredient, a plant disease control method comprising the step of treating the composition to the plant and a plant disease control agent comprising the composition.

Description

Streptomyces griseus BIG105 for controlling plant diseases and uses approximately

The present invention is streptomyces griesus having a plant disease control effect Regarding the strain VIII105 and its use, more specifically Ralstonia solanacearum ), Phytophthora capsici ), Sclerotium ( Sclerotium) cepivorum ), Rhizoctonia solani , Fusarium oxysporum) or scale Klee Loti Nias Cle to room thio (Sclerotinia sclerotiorum) Streptomyces draw with antibacterial activity against a three-house (Streptomyces griseus ) It relates to a BIG105 strain, a plant disease control composition comprising the strain as an active ingredient, a plant disease control method comprising the step of treating the composition to the plant and a plant disease control agent comprising the composition.

Blue-green disease is caused by the bacterial soil pathogen Ralstonia solanacearum and is the most damaging pathogen in tomato farms in Korea. Pathogens are motile and can survive independently for 3 to 5 years in the soil, have a high rate of disease spread at high temperatures, and when infected with a pathogen, plants wither within a short period of time. Blue-green disease is a disease that is transmitted by soil. Pathogens that invade through the roots spread through the catheter and spread throughout the catheter and damage the wood, killing the plant. In winter or when there is no host plant, in plant residues or weeds, It survives effectively and overwinters, causing illness. The path of disease spreads every year by the introduction of pathogens through soil or dust attached to farm machinery used by affected farms or through contaminated rivers, rivers and waterways. Infected fields are rapidly spread to nearby plants due to surface water flow due to irrigation or rain, or the flow of water between the irrigation materials and the soil in the facility house, which is more severe than other soil diseases.

In farms with dementia, there is no control method, so chemical chemicals are irrigated in the soil, pesticides registered in fruit trees or rice are frequently irritated, and misunderstandings are caused by local diseases, increasing the number of pesticide spraying. . In diseased soils, it is necessary to do fallow for a certain period of time or to plant crops that are not susceptible to blue-green germs. This situation is difficult to practice a lot. Currently, there are 1 item of fumigation treatment, but the drug registered in Korea for the control of cyanosis disease takes about 1 month, and when it is sprayed in a house with poor ventilation, there is a concern about human risks, vinyl coating, treatment time to secure the temperature. It is difficult to use in farms due to the limitation of the. As a result, almost all farmhouses have been burdened with production costs by purchasing seedlings by combining grafted seedlings with resistant seedlings due to concerns about cyanosis.

The research on the control of cyanosis has studied the methods of controlling chemical pesticides, antimicrobial substances and soil fumigants, but effective control methods are not known. Because chemical pesticide control is not effective, the amount of chemicals adsorbed on organic matter or clay in soil is limited to contact with pathogens, and microorganisms and drug-resistant strains are released by using processed chemicals as nutrients. It is known that the main cause is the half-life of the drug. As a research field that can replace the limitation of soil pest control of chemical pesticides, biological control methods using antagonistic microorganisms are being studied. Antagonist microorganism refers to a species that has a function of inhibiting pathogen function while inhabiting soil. Soils with abundant and abundant distribution of antagonist microorganisms do not cause disease even if they grow the same crop or artificially inoculate pathogens. There the mass cultivation of these microorganisms antagonistic be sprayed on the soil by giving control soil disease in the developed countries in agricultural soil bacillus for controlling fungal disease (Bacillus spp.), Pseudomonas species (Pseudomonas spp.), Trichoderma spp., non-pathogenic strains and yeasts are formulated and sold as microbial pesticide products. In order to develop microbial pesticides for soil disease control, it is necessary to control the pathogens to be controlled, and to control them when treated in soil or rhizosphere, and to secure antagonistic microorganisms that can survive without killing during the formulation process.

Korean Patent No. 10-0634866 discloses a strain of Ulrocradium atrum CNU 9055 having a plant fungal control effect.

The present invention is derived from the above demands, and the present inventors have found that Streptomyces griesus is isolated from soil. The present invention was completed by confirming that the BIG105 strain has inhibitory activity against various phytopathogenic bacteria.

In order to solve the above problems, the present invention Ralstonia solanacearum ( Ralstonia solanacearum ), phytophthora capsaish ( Phytophthora capsici ), Sclerotium ( Sclerotium) cepivorum , Rhizoctonia solani ), Fusarium oxysporum oxysporum ) or sclerotinia sclerotinia sclerotiorum) Streptomyces draw with antibacterial activity against three mouse (Streptomyces griseus ) Provide strains.

In addition, the present invention provides a composition for controlling plant diseases comprising the strain as an active ingredient.

The present invention also provides a plant disease control method comprising the step of treating the composition to the plant.

The present invention also provides a plant disease control agent comprising the composition.

Streptomyces griseus of the present invention BIG105 strains are expected to be useful in controlling plant diseases because they have inhibitory activity against various plant disease-causing bacteria. Since the plant disease control agent of the present invention is derived from microorganisms, it is environmentally friendly and has high safety.

1 shows Streptomyces gliseus BIG105 granular water hydrate (blue coal) showed mycelial growth inhibitory effect against red pepper blight.
2 is Streptomyces gliseus BIG105 granular water hydrant (blue coal) showed mycelial growth inhibitory effect against garlic black rot bacillus.
3 shows Streptomyces gliseus BIG105 granular watering agent (blue coal) showed mycelial growth inhibitory effect against grass lysatonia blight.
4 shows Streptomyces gliseus BIG105 granular water hydrate (blue coal) showed mycelial growth inhibitory effect against tomato wilted germ.
5 shows Streptomyces gliseus BIG105 granular water hydrant (blue coal) shows the inhibitory effect on mycelial growth against ginseng bacteria.

In order to achieve the object of the present invention, the present invention is Ralstonia solanacearum ), Phytophthora capsici ), Sclerotium ( Sclerotium) cepivorum ), Rhizoctonia solani , Fusarium oxysporum) or scale Klee Loti Nias Cle to room thio (Sclerotinia sclerotiorum) Streptomyces draw with antibacterial activity against a three-house (Streptomyces griseus ) Provides a BIG105 strain.

Streptomyces the Streptomyces griseus ) BIG105 strain is one of the bacteria isolated from soil Ralstonia solanacearum ), Phytophthora capsici ), Sclerotium cepivorum , Rhizoctonia solani ), Fusarium oxysporum oxysporum ) or sclerotinia sclerotiorum ( Sclerotinia sclerotiorum ) is a bacterium with inhibitory activity selected. Streptomyces the Streptomyces griseus ) BIG105 was deposited with Korea Research Institute of Bioscience and Biotechnology on May 27, 2010 (Accession No .: KCTC 11703BP).

In addition, the present invention provides a composition for controlling plant diseases comprising the strain as an active ingredient. Preferably, the plant disease may be, but is not limited to, red pepper disease, pepper blight, garlic black rot fungal disease, grass lysonia blight, tomato wilted disease or ginseng fungal disease.

The composition for controlling plant diseases according to the invention is prepared, for example, in the form of directly sprayable solutions, powders and suspensions or in highly concentrated aqueous, oily or other suspensions, dispersions, emulsions, oily dispersions, pastes, dusts, spraying substances or granules. It may be, but is not limited thereto.

The composition for controlling plant diseases of the present invention can be formulated in various forms. The preparation may be prepared, for example, by adding a solvent and / or a carrier. Often, inert additives and surface-active materials, such as emulsifiers or dispersants, are mixed into the formulation. Suitable surface-active materials include, but are not limited to, aromatic sulfonic acids (e.g., lignosulfonic acid, phenol-sulfonic acid, naphthalene- and dibutylnaphthalenesulfonic acid), fatty acids, alkyl- and alkylarylsulfonates, alkyl lauryl ethers, A salt of a fatty alcohol glycol ether, a sulfonate naphthalene and derivatives thereof, a condensate of formaldehyde, a condensate of naphthalene or naphthalene sulfonic acid, a phenol and a formaldehyde Condensates, polyoxyethylene octylphenol ether, ethoxylated isooctyl-, octyl- or nonylphenol, alkylphenyl or tributylphenyl polyglycol ether, alkylaryl polyether alcohol, isotridecyl alcohol, fatty alcohol / ethylene oxide condensate , Ethoxylated castor oil, polyoxyethylene alkyl ether or polyoxypropylene, lauryl alcohol Polyglycol ether acetate, sorbitol esters, lignin-sulfite waste liquid, or may be a cellulose, are not limited.

Suitable solid carrier materials are in principle all porous and agriculturally acceptable carriers, for example mineral earth (such as silica, silica gel, silicates, talc, kaolin, limestone, lime, chalk, bowls, loess, clays, Dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate, magnesium oxide, ground synthetic materials, fertilizers (such as ammonium sulfate, ammonium phosphate, ammonium nitrate, urea), vegetable products (such as grain flour, bark flour, wood meal) And nut shell powder) or cellulose powder, but is not limited thereto. The solid carrier may be used alone or in combination of two or more.

Plant disease control composition of the present invention can be used for watering, foliar spraying, seed disinfection or farm equipment disinfection, but is not limited thereto.

The composition for controlling plant diseases of the present invention is Streptomyces grossus ( Streptomyces) as an active ingredient griseus ) BIG105 strains may be used alone or in combination with two or more other antifungal, antibacterial or antiviral substances.

The composition for controlling plant diseases of the present invention may be mixed with a diffusing agent, a penetrating agent, or a surfactant to enhance crop absorption and effect.

The present invention also provides a plant disease control method comprising the step of treating the composition to the plant. Preferably, the plant disease may be, but is not limited to, red pepper disease, pepper blight, garlic black rot fungal disease, grass lysonia blight, tomato wilted disease or ginseng fungal disease. The plant disease control composition may be used by dipping, irrigation, spraying, spraying, spraying, spraying or pouring. Preferably, the composition can be treated in parallel with the immersion treatment and irrigation treatment in the plant. The mode of use depends on the intended purpose, and in all cases the distribution of the composition according to the invention should be as fine and uniform as possible.

The present invention also provides a plant disease control agent comprising the composition. Preferably, the plant disease may be, but is not limited to, red pepper disease, pepper blight, garlic black rot fungal disease, grass lysonia blight, tomato wilted disease or ginseng fungal disease.

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

Example  1: Isolation and Identification of Antagonistic Strains

Streptomyces (Streptomyces) was formed from the aerial hyphae lock kudeu chitin agar (Lockood's chitin agar) medium and preserved by the separation of pure strains that have chitin resolution. S. griseus , whose antagonism has been confirmed against cyanogen bacillus BIG105 strains are actinomycetes that form aerial mycelia and form spores on solid media, and have investigated genetic flexibilities with previously reported strains. In order to analyze the 16S DNA sequence, DNA was extracted, 16S DNA PCR amplification was performed, and the amplified PCR product was purified to analyze the base sequence. The analyzed sequencing (1,213 bp) was a strain belonging to a phylogenetic group including the species of the genus Streptomyces grise as a result of homology search in the databases of DDBJ / NCBI / Genebank and Ribosomal Database Project II. Streptomyces griseus ) was identified as Streptomyces griseus with a 99.83% flexibility relationship with ATCC 25497 ^T (D63872).

Example  2: Streptomyces Gryeus BIG105 Antimicrobial substances Production capacity

Streptomyces Griseus BIG105 had low antimicrobial production in nutrient media such as PD broth and nutrition broth, but produced antimicrobial activity against blue coliform bacteria when cultured in medium containing inorganic nutrients. Streptomyces Griseus The antibacterial material produced by BIG105 was not purified by organic solvent, but was adsorbed on Amberite IRC-50 and extracted with sulfuric acid solution. Purified antimicrobial showed similar antimicrobial activity to oxolinic acid.

Example  3: Streptomyces Gryeus BIG105  By treatment Blue and white bacteria  Density change

The soil collected was planted in mid-May and diseased at the end of June, and the soil from which the tomatoes were extracted was collected at a high concentration of 100 cfu per 1g of soil. As a result of investigating the density of pathogens during the port experiments, soil density was increased over time, and after 26 days, the density increased to 1,000,000 to 1,500,000 cfu / g. In the Streptomyces glisus BIG105 treated group, the pathogen density initially increased, but the density decreased with time, and after 12 days, the density decreased, and after 26 days, it was lower than the initial density (Table 1).

Density Changes of Blue Coliform Bacteria in Soils Caused by Antagonist Treatment (log cfu / g.Soil) Processing contents Before inoculation of antagonists 5 days after treatment 12 days after treatment 19 days after treatment 26 days after treatment Streptomyces Griseus BIG105 1.63 3.75 3.13 3.00 1.25 No treatment 2.00 2.88 4.38 4.63 6.41

Example  4: Streptomyces Gryeus BIG105  Investigation of control effect by cell treatment

Test farm packaging selection and packaging management

Greenhouse disease control farm packaging was selected from two farms that gave up farming due to severe disease in 2005 and each house (150 pyeong) was selected for antagonistic bacteria (Streptomyces griesus). BIG 105) was carried out to test the disease control effect. Fertilizer management was carried out after spraying 800 kg compost per 300 pyeong, fertilizer was sprayed with the amount of ingredients (N-20.4 kg, P-10.3 kg, K-12.2 kg), boron 2 kg, slaked lime 120 kg.

Formal and Streptomyces Gryeus BIG105  Cell treatment

The antagonistic treatment was managed so that the seedlings and grafted seedlings purchased from the nursery were allowed to soak in the antagonistic suspension when the seedlings were soaked by reducing the irrigation volume at least two days before the formulation. Streptomyces griseus in a container for seedlings BIG105 cells were prepared by diluting 1,000-fold and then immersed. The irrigation treatment was performed Streptomyces griesus diluted 1,000-fold at intervals of 7 days after the 10th day. Dilutions of BIG105 cells and chitin were irrigated three times at 5-day intervals, 150 mL per aeration.

Blue and white disease control effect

Streptomyces greece The effect of BIG105 cell treatment was tested. The untreated group could not be harvested at all by killing within 20 days of its establishment, but Streptomyces grieseus In the BIG105 cell treatment group, the control effect was different according to the number of treatments and the formulation method. The test group, which was immersed once in the regular season, was soaked with 12.5% of disease occurrence in the 4-stage harvester. In the test group immersed at the formal and irrigation at 5-day intervals, the disease was soaked with 7.5% of the 5-stage harvester. The test group irrigated with chitin at the time of initial irrigation was able to harvest tomatoes by 2.3% until the end of harvest (Table 2). The grafted seedlings, which were exposed to the resistant tree, were soaked with a 25.7% incidence of disease during the three-stage harvesting period.

On-site field trials of disease-damaged farmers Processing contents Onset time (harvest number) ^a Incidence (%) Streptomyces Griseus BIG105 dipping 4 stage 12.5 Streptomyces Griseus BIG105 3 times irrigation 5 stage 7.6 Streptomyces Griseus BIG105 + chitin (three times irrigation) 6 steps 2.3 No treatment 0 stage 100

^a point of investigation: when to harvest

On-site field trials of disease-damaged farmers (grafted to mid-July) Processing contents Onset time (harvest number) ^a Incidence (%) Streptomyces Griseus BIG105 Immersion 7-speed 3.5 Streptomyces Griseus BIG105 3 times irrigation 7-speed 0 Streptomyces Griseus BIG105 + chitin (three halls) 7-speed 0.2 No treatment 3rd stage 25.7

^a point of investigation: when to harvest

This farmhouse is cultivated in winter with triple warm house, water film and hot air facilities. In 2004, the damage of blue-green disease was severe, and in 2005, the incidence of blue-green disease was reduced by streptomyces glisus BIG105 cell treatment. Was 0.2%.

Example  5: soil treatment Granulation , For watering treatment Granular Water Wetting Agent  System treatment effect

The microbial agent for controlling blue-green disease developed in the present invention has excellent antagonism against blue-green bacteria. BIG105 was used as a starting material, and two types of products were commercialized. Granule Streptomyces Griseus Formulated with nutrients that help BIG105 settle well in soil, Streptomyces griesus in soil Antagonism of BIG105 lowers the density of cyanogen bacteria in soil. Granular moisturizer is Streptomyces griesus BIG105 is formulated with a granular watering agent to prevent clogging of filters and drip tapes. When irrigated during tomato growing, it setstles in the rhizosphere and prevents the invasion of cyanosis. Therefore, in order to control the dementia disease, it is necessary to spray the granules to the front during tillage and to periodically irrigate the granular moisturizer during the cultivation period. In the system treatment method, spray the granules 30kg per 10a to the whole field when composting. If the field is too dry, sprinkle the granules and irrigation with a fountain lake to supply water. During the planting period, the granular water hydrant is suspended in water by 300g per 10a and watered with a drop tape. The number of irrigation was performed three times at 10-day intervals to prevent pathogens from penetrating into the soft areas formed when the shoots emerged from the surface layer. Streptomyces Griseus In the treatment group treated with BIG105 granules, 87% of the dementia showed control effect, and Streptomyces griesius BIG105 granular watering agent confirmed about 78% of the dementia control effect (Tables 4 and 5).

Streptomyces Griseus Effect of BIG105 granulation treatment on blue and white diseases (after 112 days) Processing contents Byeong stock rate (%) The control
(%) repeat repeat repeat Average Streptomyces Griseus BIG105 formulation 16.0 4.0 12.0 10.7 87.7 No treatment 100 74.0 88.0 87.3 -

Streptomyces Griseus Effect of BIG105 granular wetting agent on the control of dementia (after 146 days) Processing contents Byeong stock rate (%) The control
(%) repeat repeat repeat Average Streptomyces Griseus BIG105
Granular Water Wetting Agent 15.0 12.0 13.0 13.3 77.9 No treatment 76.0 57.0 48.0 60.3 -

Example  6: Streptomyces Gryeus BIG105 Granular watering agent  Antimicrobial Activity against Phytopathogenic Fungi

Antimicrobial Activity Assay for Plant Pathogens

Streptomyces Griseus In order to examine the antimicrobial activity of the phytopathogenic fungi of the granular water soluble agent formulation based on BIG105, phytopathogenic fungi were inoculated on the PDA (patato dextrose agar) medium containing the formulation to check the mycelial growth. Assay bacteria used in the present invention were distributed at the Korea Agricultural Microbial Resources Center and described in Table 6. To compare the antimicrobial activity with conventional chemical pesticides, the antimicrobial activity against phytopathogenic fungi was compared by measuring the length of mycelial growth in the mixed medium and the control medium mixed with the control agent described in Table 6 It was.

Phytopathogenic fungi assays and controls Plant disease Test bacteria Reference drug Pepper Plague Phytophthora ( Phytophthora) capsici ) KACC 40157 Daisen M 45 Garlic black rot bacteria Sclerotium Sclerotium cepivorum ) KACC40582 Horikour Grass Raiztonia Blight Rhizoctonia solani ) AG2-2 (Ⅳ) KACC 40132 Montserlen Tomato withering bottle Fusarium oxysporum ) KACC41088 Daisen M45 Ginseng Sclerotinia sclerotiorum BIG30917 Polyoxins

In pepper blight  Antimicrobial Activity Assay

Streptomyces griseus developed in the present invention Inhibition of mycelial growth of red pepper germ bacterium was investigated in the mixed medium containing BIG105-based granular water hydrant (blue solid charcoal). Mycelial growth of pepper blight bacteria was suppressed 100% in mixed medium diluted with blue-green coal at 1,000-fold concentration, and mycelial growth inhibition effect was better than that of Daisen M45, a control pesticide specialized in pepper blight bacteria. The difference was also significant (Figure 1 and Table 7).

To garlic black rot bacteria  Antimicrobial Activity Assay

Streptomyces griseus developed in the present invention Inhibition of mycelial growth of garlic black rot bacillus bacteria in mixed medium containing BIG105-based granular water hydrate (blue coal) was significantly suppressed in the culture of garlic black rot bacteria. . Mycelial growth of garlic black rot bacteria was suppressed 100% in the mixed medium diluted chunggotan to 4,000-fold concentration, and mycelial growth was suppressed against garlic black rot bacteria than the control drug Horicure, a specialized pesticide of garlic black rot bacteria. The effect was excellent and the difference was also remarkable (FIG. 2 and Table 7).

Grass Raiztonia On blight  Antimicrobial Activity Assay

Streptomyces griseus developed in the present invention As a result of investigating the mycelial growth inhibition of the same type of grassworm fungi as grasshopper fungus, mixed medium mixed with granular water hydrate (blue coal) based on BIG105, grass lysatonia blight in medium containing blue coal Mycelial growth of bacteria was significantly suppressed. Mycelial growth of turf Laytononia blight bacteria was suppressed by 89% in mixed medium diluted to 2,000-fold concentration of blue algae. Inhibitory effect was excellent, the difference was also remarkable (Fig. 3 and Table 7).

Tomato withering  Antimicrobial Activity Assay

Streptomyces griseus developed in the present invention As a result of investigating the mycelial growth inhibition of tomato rot fungus in a mixed medium containing BIG105 based granular water hydrate (blue coal), mycelial growth of tomato wilted fungus was remarkably suppressed in the medium mixed with blue ethanol. Mycelial growth of tomato wilted pathogens was suppressed 100% in mixed medium diluted to 1000-fold concentration, and mycelial growth inhibitory effect on tomato wilted pathogens was better than that of Daisen M45, a control agent of tomato wilted pathogens. The difference was also noticeable (Figure 4 and Table 7).

In ginseng nucleus  Antimicrobial Activity Assay

Streptomyces griseus developed in the present invention As a result of investigating the mycelial growth inhibition rate of ginseng microbial bacteria in mixed medium containing BIG105-based granular water hydrant (blue coal), the mycelial growth of ginseng microbial bacteria was significantly suppressed in the medium mixed with bluecotan. Mycelial growth of ginseng bacteria was inhibited by 83.1% in mixed medium diluted to 1000-fold concentration of blue solid, and mycobacterial growth inhibition effect was better than that of polyoxin, a reference pesticide specialized in ginseng bacteria. The difference was also remarkable (FIG. 5 and Table 7).

Comparison of Mycelial Growth of Phytopathogenic Fungal Test Bacteria in Granular Water-Soluble (Blue High Coal) Mixed Medium Test bacteria Cheonggotan mixed medium Comparator mixed medium Mycelial growth length (mm) % Inhibition Mycelial growth length (mm) % Inhibition Phytophthora capsici KACC 40157 0.0 100 40.0 0 Sclerotium cepivorum KACC40582 0.0 100 37.0 9.3 Rhizoctonia solani ) AG2-2 (Ⅳ) KACC 40132 6.8 89.0 37.0 9.3 Fusarium oxysporum KACC41088 0.0 100 35.40 8.3 Sclerotinia sclerotiorum BIG30917 6.75 83.1 38.25 4.4

Korea Biotechnology Research Institute KCTC11703BP 20100527

Claims (2)

delete Streptomyces griseus (Streptomyces griseus) BIG105 strain (Accession Number: KCTC 11703BP) comprising a pepper blight or garlic black rot fungal nuclear disease control composition comprising as an active ingredient.

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