KR101695916B1 - Streptomyces badius SF7B6 strain having antimicrobial activity against pathogen for plant fungal disease isolated from flower of strawberry and uses thereof - Google Patents

Abstract

The present invention relates to a Streptomyces Vadius SF7B6 strain having an antimicrobial activity against a fungal pathogenic fungus isolated from a strawberry flower, and its use. In the present invention, a Streptomyces buddusus SF7B6 strain isolated from a strawberry flower is a bacterium, Thysseneria (Botrytis cinerea) Or Cladosporium cladosporioides (Cladosporium cladosporioides), And it was confirmed that the occurrence of gray mold and strawberry flower fungi was reduced. Therefore, the present invention is not limited to the Streptomyces bodyus (Streptomyces badius) The SF7B6 strain can be used as an environmentally friendly biological pesticide which does not have the danger of human toxicity due to environmental pollution and pesticide residues caused by side effects of pesticides and has an effect of controlling plant fungal diseases, There will be.

Description

Streptomyces badus SF7B6 strain having antimicrobial activity against a fungal pathogenic fungus isolated from a strawberry flower and a use thereof United States Patent Application 20060135899 Kind Code:

The present invention relates to a Streptomyces Vadius SF7B6 strain having antimicrobial activity against a fungal pathogenic fungus isolated from a strawberry flower and a use thereof, and more particularly, to a Streptomyces bodyus SF7B6 strain having an antimicrobial activity against a plant fungal pathogen, Streptomyces The present invention relates to a method for producing a plant fungal disease controlling composition comprising the step of cultivating the above strain and a method for producing the same, To a plant or soil, comprising the step of treating a plant fungal disease.

Crop disease caused by phytopathogenic fungi is a very limiting factor in the stable production of crops, along with pests and weeds. Among the many phytopathogenic fungi, Botrytis cinerea forms sclerotia and is a soil-infectious disease causing serious damage to various crops. These phytopathogenic fungi exist in sclerotial state and are able to survive in harsh environments such as high temperature and low temperature, and it is known that sclerotia is the primary infectious agent for plants every year. These sclerotiies can survive in diseased plant tissues or in poor environmental conditions in the soil, and are resistant to degradation and inhibition by antagonistic microorganisms, so it is difficult to expect effective control. Methods for controlling these pathogenic fungi include physical methods to improve the cultivation environment, chemical methods using microbicides, and microbiological methods using microorganisms. However, physical control alone can not be expected to have a great effect. Chemical control has recently led to the advent of drug-resistant pathogens, resulting in a significant deterioration in the pest control effect, and serious environmental pollution caused by large-scale application of chemical pesticides. In addition, the use of chemical synthetic pesticides is increasingly restricted due to toxicity to humans and animals, residual toxicity in soil and crops, environmental pollution caused by packaging containers, etc. In advanced countries, And the use of pesticides is being restricted by strict regulations.

Gray mold fungus is a global botanical disease that occurs worldwide and causes enormous economic damage before and after harvesting of various crops including fruit and vegetables. The causative bacteria of gray mold ( Botrytis Cinerea occurs in all parts of crops such as leaves, stems, flowers, fruits and roots, and is particularly damaging to crops grown in institutional houses. The causative agent of gray mold causes a secondary infection in aged tissues, tissues damaged by wounds or other diseases due to the nature of pathogen and secondary life. The causative bacteria of gray mold are able to survive in chemical pesticides and various unsuitable environments by forming sclerotial sclerotia, and can maintain pathogenicity for many years in soil and dead plant debris. They can grow at a temperature as low as 4 ° C. If they can not ventilate due to the temperature drop at night in a facility house in winter, they will be exposed to low temperature and high humidity conditions, resulting in gray mold fungi. It is relatively easy to obtain a resistance to a drug and thus it is difficult to control the drug.

In addition, strawberry flower fungus disease, once it occurs, is spread rapidly due to wind or bees, and it is difficult to control, and it causes anomalies by interfering with flower modification. In particular, the causative bacteria of Staphylococcus aureus ( Cladosporium cladosporioides ) is 20-25 ° C. It tends to occur in a house with high humidity and condensation, or in an environment with low sunshine, and can also grow in dead plants, soil, and organic matter. At present, the method of controlling the strawberry flower fungus disease is to reduce the humidity in the house and to prevent the environment-friendly scion registered in strawberry powdery mildew disease or anthrax disease before the blooming period, and the medicines with little influence on the germination of the pollen. Therefore, there is a need to develop an eco-friendly pesticide that does not cause residual toxicity and environmental pollution.

Korean Patent No. 1481897 discloses Streptomyces sp. BS063 and plant disease control using the same. Korea Patent No. 1403304 discloses Streptomyces strains JS (II) -10 strain, A microorganism preparation for controlling a plant gray mold disease and a method for controlling a gray mold disease have been disclosed, but Staphylosmiths bodyfus SF7B6 strain having antimicrobial activity against a plant fungal pathogen isolated from a strawberry flower of the present invention and its use Have not been disclosed.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-described needs, and it is an object of the present invention to provide a Streptomyces budesus SF7B6 strain isolated from the strawberry flower of the present invention is Botrytis cinerea ) or Cladosporium cladosporioides to reduce the occurrence of gray mold and strawberry flower fungi. Thus, the present invention has been accomplished by confirming the control effect of Streptomyces albus SF7B6 on plants, in particular, strawberry fungal diseases.

In order to accomplish the above object, the present invention provides a method of treating Streptomyces vesicularis badius ) SF7B6 strain.

In addition, the present invention is the Streptomyces mouse body (Streptomyces badius) provides a composition for controlling plant fungal disease control, including SF7B6 strain or culture thereof as an active ingredient.

In addition, the present invention is the Streptomyces mouse body (Streptomyces badius) provides a method for preparing a composition for controlling plant fungus disease which comprises the step of culturing the strain SF7B6.

In addition, the present invention is the Streptomyces mouse body (Streptomyces treating a plant fungal disease comprising treating a plant or soil with an effective amount of a badius SF7B6 strain or a culture thereof.

In the present invention, the Streptomyces bodyus SF7B6 strain isolated from the strawberry flower is a Botrytis < RTI ID = 0.0 > cinerea ) or Cladosporium cladosporioides to reduce the occurrence of gray mold and strawberry flower fungi. Thus, Streptomyces process of the present invention my mouse body (Streptomyces badius ) SF7B6 strain can be used as environmentally friendly biological pesticide which does not have the danger of human toxicity due to environmental pollution and residual pesticide caused by the side effect of pesticide and has a controlling effect against plant fungal disease, It will be possible.

Figure 1 shows the date of strawberry flower collection (A) and the incidence of strawberry gray mold (B) in the strawberry vinyl house plantation.
Figure 2 shows the result of analysis of the metagenome from strawberry flowers.
Fig. 3 shows the results of 16S rRNA sequences of Streptomyces badius strain SF7B6 isolated from strawberry flowers.
Fig. 4 shows the antimicrobial activity of the strain Streptomyces badius SF7B6 isolated from strawberry flowers against the causative bacteria of strawberry gray mold. B. cinerea is a causative agent of Staphylococcus aureus fungi, and SF7B6 is Streptomyces bodyus SF7B6 strain.
FIG. 5 shows the antimicrobial activity of the Streptomyces badius strain SF7B6 isolated from strawberry flowers against the causative bacteria of Staphylococcus aureus. C. cladosporioides is a causative organism of Staphylococcus aureus and SF7B6 is a Streptomyces Vadius SF7B6 strain.

In order to achieve the present invention, the present invention is Streptomyces mouse body (Streptomyces with antibacterial activity against plant fungal disease pathogens badius ) SF7B6 strain.

The Streptomyces mouse body (Streptomyces badius ) SF7B6 isolates were isolated from strawberry flowers, and strawberry fungal pathogen pathogens (Staphylococcus aureus, Botrytis cinerea ) and Strawberry flower mold causative bacteria ( Cladosporium cladosporioides ) as a strain with excellent antimicrobial activity. The Streptomyces mouse body (Streptomyces badius ) SF7B6 strain was deposited at the Korean Culture Center of Microorganisms (KCCM) on June 3, 2015 (KCCM 11702P).

In the strain according to an embodiment of the present invention, the plant pathogenic fungus pathogen may be a causative agent of a gray mold or a flower mold fungus of Staphylococcus aureus , and preferably Botrytis cinerea cinerea , or Cladosporium cladosporioides . The term " cladosporium cladosporioides "

In addition, the present invention is the Streptomyces mouse body (Streptomyces badius) provides a composition for controlling plant fungal disease control, including SF7B6 strain or culture thereof as an active ingredient. The composition for controlling plant fungal diseases may be used as a microbial pesticide.

In the composition according to an embodiment of the present invention, the plant fungal disease may be a gray mold or a strawberry flower fungal disease, and the gray mold disease may be caused by a disease such as cucumber, strawberry, grape, pepper, lettuce, tomato, peach, It may be a gray mold, preferably a strawberry gray mold, but is not limited thereto.

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

The composition for controlling plant fungal diseases of the present invention can be formulated into various forms. The preparation can 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 ethers, ethoxylated isooctyl-, octyl- or nonylphenol, alkylphenyl or tributylphenyl polyglycol ethers, alkylaryl polyether alcohols, isotridecyl alcohol, fatty alcohol / ethylene oxide condensates , 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 comprise an agriculturally acceptable carrier such as mineral earths such as silica, silica gel, silicate, talc, kaolin, limestone, limestone, chalk, Fertilizers such as ammonium sulfate, ammonium phosphate, ammonium nitrate, urea, vegetable products such as cereal flour, bark powder, wood meal, And nut shell powder), or cellulose powder. The solid carrier may be used alone or in combination of two or more.

For fungal plant disease controlling composition of the present invention as an active ingredient Streptomyces mouse body (Streptomyces badius ) SF7B6 strain alone or in combination with two or more different antifungal, antibacterial or antiviral substances.

The plant fungal disease controlling composition of the present invention can be mixed with a dispersing agent, a penetrating agent, or a surfactant to enhance the absorption and effect of plants.

In addition, the present invention is the Streptomyces mouse body (Streptomyces badius) provides a method for preparing a composition for controlling plant fungus disease which comprises the step of culturing the strain SF7B6.

Any method known in the art can be used for culturing the strain, and the method is not particularly limited.

In addition, the present invention is the Streptomyces mouse body (Streptomyces treating a plant fungal disease comprising treating a plant or soil with an effective amount of a badius SF7B6 strain or a culture thereof.

As a way to control the fungal plant disease is the Streptomyces mouse body (Streptomyces badius ) SF7B6 strain in a plant or soil, but is not limited thereto.

The " effective amount " of the present invention means that the control composition is uniformly diluted with water to control the plant fungal disease, in an amount sufficient to produce beneficial or desired results, and then sprayed onto the plant and arable land using a suitable spraying device such as a power sprayer can do. When the wettable powder or liquid wettable powder of the present invention is diluted in water, the concentration of the wettable powder or the liquid wettable powder is preferably in the range of 10 ⁵ to 10 ¹⁰ cfu / ml, preferably 10 ⁸ cfu / ml or less so that the effective ingredient may be in a biologically effective range , More preferably 10 < ⁶ > cfu / ml, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Identification of the incidence of strawberry gray mold disease in strawberry vinyl house plantations

In order to confirm the incidence of gray mold of strawberry, the incidence of strawberry gray mold disease was checked every two weeks from November 14, 2013 to April 30, 2014, at strawberry vinyl house planting site in Suok- 18 straws were randomly collected in the strawberry vinyl house (FIG. 1B). The incidence of strawberry gray mold disease was investigated by infecting the gray mold fungus infected with 100 strawberry. As a result, it was confirmed that the strawberry gray mold disease occurred from the eighth time as shown in Fig. 1B.

Example  2. From strawberry flowers Meta genome ( metagenome ) analysis

In order to analyze the metagenome from strawberry flowers, the strawberry flowers collected in thirteen times were put into PBS buffer and sonicated. Then, genomic DNA was extracted using Solgent DNA prep kit (Solgent, Korea). To perform pyrosequencing, the extracted genomic DNA was stored at -80 ° C. The portion of the 16S rRNA of the bacteria was amplified and subjected to Pyrosequencing analysis in Macrogen (Korea). All base sequences were identified using the EzTaxon database (http://eztaxon-e.ezbiocloud.net/). As a result, since the density of Actinobacteria phylum rapidly drops from the 7th flower (Flower 7) where the gray mold disease starts to occur as shown in Fig. 2, the Actinobacteria gate phylum) was closely related to the occurrence of gray mold.

Example  3. Streptomyces Bodyus ( Streptomyces badius ) SF7B6  Isolation and Identification of Strain

The collected strawberry flowers were diluted with PBS buffer, sonicated, and diluted with TSA medium to isolate microorganisms. Genomic DNA was isolated to analyze the 16S rRNA sequence (SEQ ID NO: 1) in order to carry out the molecular biology identification of isolated microorganisms. Streptomyces mouse body (Streptomyces genomic DNA of the badius SF7B6 strain was extracted using CTAB (Cetyl trimethyl ammonium bromide) technique. For the sequencing analysis, the primers shown in the following Table 1, which targets the regions V1 to V3 of 16S rRNA, were used. As a result, as shown in FIG. 3, the 16S rRNA nucleotide sequence was identified as Streptomyces badius through BLAST search of NCBI, and the strain was named Streptomyces Vadius SF7B6 strain.

The primers used in the present invention primer
name SEQ ID NO: The forward direction (5'-3 ')
Primer base sequence primer
name SEQ ID NO: The reverse direction (5'-3 ')
Primer base sequence 27F 2 AGAGTTTGATCMTGGCTCAG 1492R 3 GGYTACCTTGTTACGACTT

Example  4. Streptomyces Bodyus ( Streptomyces badius ) SF7B6  Staphylococcus aureus fungal disease Botrytis cinerea ) For Road drag  Confirm

Example 1 to 3 and separated from one Streptomyces body mouse (identified Streptomyces To confirm the antimicrobial activity of the badius SF7B6 strain against the causative agents of strawberry gray mold, Botrytis cinerea , a causative organism of strawberry gray mold, was used. To verify the antimicrobial activity, strains of Streptomyces Vadius SF7B6 and Staphylococcus aureus ( Botrytis spp.) Were cultivated on a PDA (potato texturus 10 g, peptone 10 g, agar 20 g) cinerea ). At this time, incubation temperature of 27 ° C, which is the growth temperature of various pathogens, was observed in the inhibition zone. As a result, as shown in FIG. 4, the Streptomyces budesus SF7B6 strain of the present invention was found to exhibit a strong antimicrobial activity against the causative bacteria of Staphylococcus aureus.

Example  5. Streptomyces Bodyus ( Streptomyces badius ) SF7B6  Staphylococcus aureus Staphylococcus aureus Cladosporium cladosporioides ) For Road drag  Confirm

Example 1 to 3 and separated from one Streptomyces body mouse (identified Streptomyces Cladosporium cladosporioides , a causative organism of Staphylococcus aureus , was used to identify the antimicrobial activity of the badius SF7B6 strain against the causative bacteria of Staphylococcus aureus . In order to verify the antimicrobial activity, Streptomyces Vadius SF7B6 and Cladosporium spp. Were cultivated on a PDA (potato texturus 10 g, peptone 10 g, agar 20 g) cladosporioides ). At this time, incubation zone was observed after culturing at 20 ° C according to characteristics of cold bacteria. As a result, as shown in FIG. 5, it was confirmed that the Streptomyces budesus SF7B6 strain of the present invention showed a strong antibacterial activity against the causative bacteria of Staphylococcus aureus.

Korea Microorganism Conservation Center (overseas) KCCM11702P 20150603

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> Streptomyces badius SF7B6 strain having antimicrobial activity          against pathogen for plant          strawberry and uses thereof <130> PN15143 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 941 <212> DNA <213> Streptomyces badius <400> 1 ctgcccttca gagctatctg cagtcgaacg atgagccctt cngggtggat tagnggcgaa 60 cgggtgagta acacgtgggc aatctgccct tcactctggg acaagccctg gaaacggggt 120 ctaataccgg ataacactct gtcccgcatg ggacggggtt gaaagctccg gcggtgaagg 180 atgagcccgc ggcctatcag cttgttggtg gggtaatggc ctaccaaggc gacgacgggt 240 agccggcctg agagggcgac cggncacact gggactgaga cacggcccan actcctacgg 300 gaggcagcag tggggaatat tgcacaatgg gcgaaagcct gatgcagcga cgccgcgtga 360 gggatgacgg ccttcgggtt gtaaacctct ttcagcaggg aagaagcgaa agtgacggta 420 cctgcagaag aagcgccggc taactacgtg ccagcagccg cggtaatacg tagggcgcaa 480 gcgttgtccg gaattattgg gcgtaaagag ctcgtaggcg gcttgtcacg tcggatgtga 540 aagcccgggg cttaaccccg ggtctgcatt cgatacgggc tagctagagt gtggtagggg 600 agatcggaat tcctggtgta gcggtgaaat gcgcagatat caggaggaac accggtggcg 660 aaggcggatc tctgggccat tactgacgct gaggagcgaa agcgtgggga gcgaacagga 720 ttagataccc tggtagtcca cgccgtaaac gttgggaact aggtgttggc gacattccac 780 gtcgtcggtg ccgcagctaa cgcattaagt ncccgcctgg ggagtacggc cgcaaggcta 840 aactcaagga ttgacggggg cccgcacaag cagcgnagca tgtggnttat tcgacgcacg 900 cgaaaacctt accaggcttg gagcatgtgg cttaattcga c 941 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 agagtttgat cmtggctcag 20 <210> 3 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ggytaccttg ttacgactt 19

Claims (9)

Streptomyces badius strain SF7B6 (KCCM 11702P) having antibacterial activity against Botrytis cinerea and Cladosporium cladosporioides . delete delete A method for producing Streptomyces badius strain SF7B6 according to claim 1 or a method for producing the same by culturing Botrytis cinerea or Cladosporium cladosporioides A composition for controlling plant fungal diseases. delete delete A method for the treatment of plant fungi by Botrytis cinerea or Cladosporium cladosporioides , comprising the step of culturing Streptomyces badius strain SF7B6 according to claim 1, A method for producing a composition for preventing disease control. A method for the treatment of Botrytis cinerea or Cladosporium cladosporioi comprising the step of spraying an effective amount of a Streptomyces badius strain SF7B6 according to claim 1 or a culture thereof to a plant or soil, A method for controlling plant fungal diseases by Cladosporium cladosporioides . delete

Images