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KR-102962349-B1 - Bacteriophage mixture FireFighter-A for controlling fire blight or black blight

KR102962349B1KR 102962349 B1KR102962349 B1KR 102962349B1KR-102962349-B1

Abstract

One aspect relates to FireFighter-A, a bacteriophage mixture for controlling fire blight or black rot of eggplant. According to one aspect, when a cocktail (mixture) of four types of bacteriophages is applied, it was confirmed to have potent lytic activity specific to Erwinia amylovora and/or Erwinia pyrifoliae and to be stable under temperature, pH, and UV light. In addition, it was confirmed that when a cocktail (mixture) of four types of bacteriophages is applied, compared to treatment with a single bacteriophage, it prevents the development of strains resistant to a single bacteriophage, thereby being effective in controlling fire blight and/or black rot of eggplant, and thus can be utilized for the prevention or control of fire blight and/or black rot of eggplant.

Inventors

  • 노은정
  • 김벼리
  • 송수진
  • 박세창
  • 김상근

Assignees

  • 대한민국(농촌진흥청장)
  • 서울대학교산학협력단

Dates

Publication Date
20260508
Application Date
20220616

Claims (15)

  1. A bacteriophage mixture FireFighter-A (accession number: KACC 97042P) comprising Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47 having lytic activity against one or more microorganisms selected from the group consisting of Erwinia amylovora and Erwinia pyrifoliae .
  2. The bacteriophage mixture of claim 1, wherein Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47 are of the Myoviridae family.
  3. A bacteriophage mixture according to claim 1, wherein one or more bacteriophages selected from the group consisting of Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47 have lytic activity at 30°C to 50°C.
  4. A bacteriophage mixture according to claim 1, wherein one or more bacteriophages selected from the group consisting of Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47 have lytic activity at pH 4 to 9.
  5. A bacteriophage mixture according to claim 1, wherein one or more bacteriophages selected from the group consisting of Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47 have lytic activity after exposure to sunlight of 1 SUN (100 mW/ cm² ) for 3 to 9 hours.
  6. A composition for controlling one or more microorganisms selected from the group consisting of Erwinia amylovora and Erwinia pyrifoliae, comprising the mixture of claim 1.
  7. A control composition according to claim 6, wherein the composition has lytic activity against fruit tree bacterial diseases.
  8. A pest control composition according to claim 6, wherein the composition is treated at a concentration of 0.01 to 100 MOI.
  9. A control composition according to claim 7, wherein the fruit tree bacterial disease is one or more selected from the group consisting of fire blight and black rot of branches.
  10. A pest control composition according to claim 6, wherein the composition further comprises an excipient.
  11. A pest control composition according to claim 6, wherein the composition is one or more formulations selected from the group consisting of wettable powders, granular wettable powders, liquid wettable powders, liquids, aqueous solutions, water-soluble granules, and encapsulating agents.
  12. A pest control composition according to claim 6, wherein the composition is one or more selected from the group consisting of a trunk injection composition, a biological agent composition, a fertilizer composition, a nutrient composition, a plant strengthening composition, a plant protection composition, a soil conditioner composition, and a yield enhancer composition.
  13. A method for controlling fruit tree bacterial diseases, comprising the step of treating a plant or soil on which the plant is planted with the control composition of claim 6.
  14. A method according to claim 13, wherein the fruit tree bacterial disease is one or more selected from the group consisting of fire blight and black rot of branches.
  15. The method of claim 13, wherein the plant is one or more plants selected from the group consisting of apple, pear, rose, loquat, quince, apricot, peach, plum, apricot, aronia, cherry, cherry, and rose.

Description

Bacteriophage mixture FireFighter-A for controlling fire blight or black blight This relates to FireFighter-A, a bacteriophage mixture for controlling fire blight or black rot. Fire blight is one of the representative bacterial plant quarantine diseases, and * Erwinia amylovora * is known to be the causative agent. Fire blight causes the flowers, branches, and fruits of trees such as pear or apple to turn black, resulting in symptoms where they dry up and die as if burned. *Erwinia amylovora* is highly contagious because it is transmitted by insects or wind and rain and invades the host plant through flowers, nectaries, stomata, lenticels, and wounds. It is characterized by its ability to infect about 70 species of plants, including pear, apple, and apricot trees, leading to damage caused by fire blight worldwide. Erwinia pyrifoliae is the pathogen of black blight, and it causes severe necrosis in the Asian pear tree * Pyrus pyrifolia *. In 1995, black or brown streaks along the central veins and necrosis of the leaf petioles were observed on the leaves of the Asian pear variety *Shingo* in an orchard near Chuncheon. These symptoms spread to the entire branches and affected flowers and small fruits, resulting in a significant decrease in fruit yield. As a response to fire blight and black rot, methods to control the infection and spread of fire blight and black rot by reducing soil moisture and maintaining a balance of fertilizer nutrients, and methods to treat fire blight and black rot by removing infected fruit trees or blackened branches in winter when the pathogens are dormant are known, but they cannot eliminate the essential occurrence of the disease. Accordingly, the inventors studied phage therapy, a control (treatment) method using bacteriophages as a method for controlling Erwinia amylovora and/or Erwinia pyrifolia, and discovered that a novel bacteriophage or a bacteriophage cocktail (mixture) containing it exhibits strong lytic activity against Erwinia amylovora and/or Erwinia pyrifolia. Figure 1 is a figure showing the results of isolating four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47) and confirming plaques using Erwinia amylovora YKB14808, Figure 1a is a figure confirming the plaque of Erwinia phage Fifi451, Figure 1b is a figure confirming the plaque of Erwinia phage Fifi318, Figure 1c is a figure confirming the plaque of Erwinia phage pEa_SNUABM_27, and Figure 1d is a figure confirming the plaque of Erwinia phage pEa_SNUABM_47. Figure 2 is a figure showing the results of morphological analysis of four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47), Figure 2a is a figure analyzing the morphology of Erwinia phage Fifi451, Figure 2b is a figure analyzing the morphology of Erwinia phage Fifi318, Figure 2c is a figure analyzing the morphology of Erwinia phage pEa_SNUABM_27, and Figure 2d is a figure analyzing the morphology of Erwinia phage pEa_SNUABM_47. Figure 3 is a figure showing the results of confirming the temperature stability of four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47), Figure 3a is a figure confirming the temperature stability of Erwinia phage Fifi451, Figure 3b is a figure confirming the temperature stability of Erwinia phage Fifi318, Figure 3c is a figure confirming the temperature stability of Erwinia phage pEa_SNUABM_27, and Figure 3d is a figure confirming the temperature stability of Erwinia phage pEa_SNUABM_47. Figure 4 shows the results of confirming the pH stability of four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47), Figure 4a shows the pH stability of Erwinia phage Fifi451, Figure 4b shows the pH stability of Erwinia phage Fifi318, Figure 4c shows the pH stability of Erwinia phage pEa_SNUABM_27, and Figure 4d shows the pH stability of Erwinia phage pEa_SNUABM_47. Figure 5 is a figure showing the results of confirming the UV stability of four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47), Figure 5a is a figure confirming the UV stability of Erwinia phage Fifi451, Figure 5b is a figure confirming the UV stability of Erwinia phage Fifi318, Figure 5c is a figure confirming the UV stability of Erwinia phage pEa_SNUABM_27, and Figure 5d is a figure confirming the UV stability of Erwinia phage pEa_SNUABM_47. Figure 6 is a phylogenetic tree showing four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi318, Erwinia phage pEa_SNUABM_27 and Erwinia phage pEa_SNUABM_47). Figure 7 is a figure showing the results of comparing the growth inhibition effects of four types of bacteriophages ( Erwinia phage Fifi451, Erwinia phage Fifi