Composition containing a chemical substance having antibacterial activity and use thereof

Abstract

The present invention relates to a composition for inhibiting the growth of fire blight bacteria, comprising cyclo(-Met-Pro) as an active ingredient.

Description

Technology Field

[0001] The present invention relates to a composition comprising a chemical substance having antibacterial activity. Background Technology

[0002] Fire blight first broke out in Korea in 2015 at an apple farm in Anseong, Gyeonggi-do, spreading to 43 farms and resulting in the burial of 68 farms. In 2016, it spread to 17 farms and resulted in the burial of 32 farms, in 2017 to 33 farms and 55 farms, and in 2018 to 67 farms and 135 farms. The number of farms affected by fire blight is increasing rapidly every year.

[0003] Overseas, it first occurred in the 1780s in apple and pear orchards in the Hudson Valley, New York, USA. Over time, as of April 2021, the fire blight strain has spread throughout the United States and Canada, affecting at least 60 countries worldwide. In Europe, the fire blight pathogen is designated as a quarantine disease under the A2 regulations of the European Plant Protection Agency.

[0004] To date, no clear cure for fire blight has been developed, and as it is a disease that causes significant human and / or economic losses by burying all host plants within a certain radius with just a single infection, it is urgent to devise measures to reduce damage to farms and national losses.

[0005] As antibiotics such as streptomycin, which are conventionally used to control fire blight, have problems such as toxicity, residue issues, and the acquisition of resistance by pathogens, there is a trend toward researching biological control methods. The problem to be solved

[0006] The objective of the present invention is to provide a composition comprising a chemical substance that inhibits the growth of plant pathogens. means of solving the problem

[0007] The present invention relates to a composition for inhibiting the growth of fire blight bacteria, comprising cyclo(-Met-Pro) as an active ingredient.

[0008] In one embodiment of the present invention, the cyclo(-Met-Pro) may be included at a concentration of 100 μg / mL to 5,000 μg / mL.

[0009] In one embodiment of the present invention, the cyclo(-Met-Pro) may have the effect of inhibiting the growth of at least one of fire blight bacteria, black rot bacteria, bacterial wilt bacteria, ulcer bacteria, or soft rot bacteria.

[0010] The present invention relates to a method for inhibiting the growth of fire blight bacteria, wherein a composition for inhibiting the growth of fire blight bacteria according to one embodiment of the present invention is used with Erwinia amylovora ( Erwinia amylovora The method includes the step of treating at least one of the surrounding area of ​​the Erwinia amylovora, the host plant of the Erwinia amylovora, or the surrounding area of ​​the host plant of the Erwinia amylovora.

[0011] The present invention relates to a composition for inhibiting fire blight, comprising Cyclo(-Met-Pro) as an active ingredient, and Erwinia amylovora ( Erwinia amylovora It has a growth inhibitory effect on ).

[0012] The present invention relates to a composition for inhibiting the growth of plant pathogens, comprising Cyclo(-Met-Pro) as an active ingredient, and Erwinia amylovora ( Erwinia amylovora ), Erwinia Piripolie ( Erwinia pyrifoliae ), Ralstonia solanacearum( Ralstonia solanacearum ), Clavibacter miciganensis ( Clavibacter michiganensis ), or Pectobacterium carotoborum ( Pectobacterium carotovorum It has a growth inhibitory effect on at least one of ).

[0013] The present invention relates to a composition for inhibiting the growth of eggplant black blight fungi, comprising cyclo(-Met-Pro) as an active ingredient.

[0014] The present invention relates to a composition for inhibiting the growth of bacterial wilt pathogens, comprising cyclo(-Met-Pro) as an active ingredient.

[0015] The present invention relates to a composition for inhibiting the growth of ulcer pathogens, comprising cyclo(-Met-Pro) as an active ingredient.

[0016] The present invention relates to a composition for inhibiting the growth of soft rot pathogens, comprising cyclo(-Met-Pro) as an active ingredient. Effects of the invention

[0017] A composition for inhibiting the growth of plant pathogens according to one embodiment of the present invention can inhibit the growth of various plant pathogens. Brief explanation of the drawing

[0018] Figure 1 is a photograph showing the results of an experiment on the growth inhibition ability of fire blight pathogens using the culture filtrate of the Cerratia rubidea AO4-P6 strain according to one embodiment of the present invention. FIG. 2 is a graph showing the degree of inhibition of fire wilt pathogen growth for each of the fractions separated according to one embodiment of the present invention. FIG. 3 is a graph showing the chromatographic results of some of the fractions according to one embodiment of the present invention. Figure 4 is a graph showing the degree of growth inhibition of fire blight pathogens by a candidate substance for inhibiting fire blight pathogens isolated according to one embodiment of the present invention. Figure 5 is a graph showing the degree of growth inhibition of black blight pathogens by a fire blight pathogen inhibitory substance isolated according to one embodiment of the present invention. Figure 6 is a graph showing the degree of growth inhibition of bacterial blight pathogens by a fire blight pathogen inhibitory substance isolated according to one embodiment of the present invention. Figure 7 is a graph showing the degree of inhibition of ulcer disease growth by a fire blight inhibitory substance isolated according to one embodiment of the present invention. Figure 8 is a graph showing the degree of growth inhibition of soft rot bacteria by a fire blight inhibitory substance isolated according to one embodiment of the present invention. Specific details for implementing the invention

[0019] Hereinafter, a composition comprising a chemical substance having antibacterial activity according to the present invention and its uses will be described in detail with reference to the attached drawings.

[0020] Prior to this, the terms used in this specification and claims shall not be interpreted as being limited to their dictionary meanings; rather, based on the principle that the inventor may appropriately define the concepts of the terms to best describe their invention, they shall be interpreted in a meaning and concept consistent with the technical spirit of the invention.

[0021] The exemplary embodiments described in the detailed description, drawings, and claims are not intended to be limiting, and other embodiments may be used, and other modifications are possible without departing from the spirit or scope of the technology disclosed herein.

[0022] Therefore, it should be understood that the embodiments described in this specification and the configurations illustrated in the drawings are merely preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention, and that various equivalents and modifications that can replace them may exist at the time of filing this application.

[0023] Those skilled in the art will readily understand that the components of this disclosure, namely those generally described herein and those depicted in the drawings, can be arranged, configured, combined, and designed in various different configurations, all of which are obviously devised and form part of this disclosure.

[0024] The present invention is characterized by providing a composition for inhibiting the growth of fire blight bacteria, a composition for suppressing fire blight, and a method for inhibiting the growth of fire blight bacteria by treating the composition for inhibiting the growth of fire blight bacteria or the composition for suppressing fire blight.

[0025] Fruit blight (or fire blight) primarily affects fruit trees belonging to the 39 genera and over 180 species of the genus Rosa, mainly Erwinia amylovora ( Erwinia amylovora It is a disease caused by the strain. The aforementioned Erwinia amylovora strain primarily overwinters in the tissues of tree trunks or branch edges, becomes active starting in the spring when flowering approaches, and transmits bacterial mucus to other plants through pollinators such as insects like bees, butterflies, and flies, birds, natural elements including wind and rain, pruning tools, and / or humans. Because it spreads easily through a wide variety of routes, it exhibits rapid transmissibility to host plants in nearby areas after the onset of the disease.

[0026] If the aforementioned strain invades and proliferates within a tree, it can infect the entire plant, including flowers, leaves, fruits, and branches, ultimately causing the entire tree to wither. In plants infected with fire blight, the leaves initially turn black along the main vein or leaf veins, and as the disease progresses, the entire leaf dries out and turns brown or black. The flowers of infected plants initially appear wet and turn a grayish-white green, turning black and drying out as the disease progresses. The fruits of infected plants begin to turn black, and white, orange, or brown bacterial exudate forms on the surface. The bark of infected branches appears wet, the surface sinks, and ulcers form; young branches rapidly wilt, turn black, and their tips bend like hooks.

[0027] The present invention provides a composition for inhibiting the growth of fire blight pathogens comprising cyclo(-Met-Pro) as an active ingredient.

[0028] The concentration of Cyclo(-Met-Pro) included as an active ingredient in the composition for inhibiting the growth of fire blight bacteria according to the present invention can be adjusted. For example, the concentration of Cyclo(-Met-Pro) may be 1 μg / mL to 10,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 10 μg / mL to 7,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 100 μg / mL to 5,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 200 μg / mL to 5,000 μg / mL.

[0029] The composition for inhibiting the growth of fire blight bacteria according to the present invention may additionally include any other component or microorganism used in the field for inhibiting the growth of fire blight bacteria in addition to Cyclo(-Met-Pro).

[0030] Furthermore, since the composition for inhibiting the growth of fire blight bacteria according to one embodiment of the present invention comprises a cyclo(-Met-Pro) that inhibits fire blight bacteria, the composition of the present invention may be used for purposes other than the composition for inhibiting the growth of fire blight bacteria disclosed above, and its name may also be appropriately changed. However, there may be differences in degree depending on the terms used, and adjustments may be made according to the details specified in the examples.

[0031] For example, the composition of the present invention comprising cyclo(-Met-Pro) as an active ingredient is a composition for inhibiting the growth of plant pathogens, a composition for suppressing plant pathogens, a composition for controlling plant pathogens, a composition for preventing plant pathogens, a plant pathogen growth inhibitor, a plant pathogen suppressor, a plant pathogen control agent, a plant pathogen preventive agent, a composition for suppressing plant diseases, a composition for controlling plant diseases, a composition for preventing plant diseases, a plant disease suppressor, a plant disease control agent, a plant disease preventive agent, a composition for inhibiting the growth of fire blight pathogens, a composition for controlling fire blight pathogens, a composition for preventing fire blight pathogens, a fire blight pathogen growth inhibitor, a fire blight pathogen suppressor, a fire blight pathogen control agent, a fire blight preventive agent, a composition for suppressing fire blight, a composition for controlling fire blight, a composition for preventing fire blight, a fire blight suppressor, a fire blight control agent, a fire blight preventive agent, Erwinia amylovora ( Erwinia amylovora It may be named a composition for inhibiting growth, a composition for inhibiting Erwinia amylovora, a composition for controlling Erwinia amylovora, a composition for preventing Erwinia amylovora, a growth inhibitor, an Erwinia amylovora inhibitor, an Erwinia amylovora control agent, an Erwinia amylovora preventive agent, etc.

[0032] In one embodiment of the present invention, the growth of fire blight bacteria can be inhibited by treating at least one of the fire blight bacteria, the surrounding area of ​​the fire blight bacteria, the host plant of the fire blight bacteria, the surrounding area of ​​the host plant of the fire blight bacteria, or the soil with the composition for inhibiting the growth of fire blight bacteria.

[0033] The host plants of the fire blight pathogen according to one embodiment of the present invention include fruit trees belonging to the 39 genera and 180 species of the Rosaceae family. Examples include apple trees, pear trees, quince trees, apricot trees, peach trees, plum trees, apricot trees, aronia, raspberry, spirea, roses, rowan trees, quince trees, cherry trees, strawberries, etc.

[0034] The present invention relates to a composition for inhibiting the growth of fire blight bacteria comprising cyclo(-Met-Pro) as an active ingredient, wherein the fire blight bacteria (Erwinia amylovara ( Erwinia amylovora The present invention provides a method for inhibiting the growth of fire blight bacteria, comprising the step of treating at least one of the surrounding area of ​​the fire blight bacteria, the host plant of the fire blight bacteria, or the surrounding area of ​​the host plant of the fire blight bacteria. Referring to the prior art, the method for inhibiting the growth of fire blight bacteria according to one embodiment of the present invention may be named a method for inhibiting fire blight bacteria, a method for inhibiting the growth of plant pathogens, a method for inhibiting fire blight, a method for inhibiting plant diseases, etc., but is not limited thereto.

[0035] The term “surrounding area” refers to the range in which the composition of the present invention can exert its effects, and may include the vicinity of the fire blight bacteria to which the composition is treated, the vicinity of the host plant of the fire blight bacteria, and both the above-ground and underground areas near the host plant.

[0036] For example, the above surrounding area may be an area within about 2 m, about 1 m, about 70 cm, about 50 cm, about 25 cm, about 10 cm, or about 5 cm around the fire blight bacteria, the host plant of the fire blight bacteria, a part of the host plant, or an area around the fire blight bacteria and the host plant.

[0037] According to one embodiment of the present invention, the part of the host plant is not limited and may include at least one of the regions recognized as part of the plant, such as the leaves of the host plant, the stem of the host plant, the roots of the host plant, the fruits of the host plant, and the flowers of the host plant.

[0038] A method for inhibiting the growth of fire blight bacteria according to one embodiment of the present invention may be applied under various conditions. For example, a method for inhibiting the growth of fire blight bacteria may use the composition for inhibiting the growth of fire blight bacteria of the present invention against fire blight bacteria (Erwinia amylovora ( Erwinia amylovora It can be provided to at least one of the surrounding area of ​​the fire blight pathogen, the host plant of the fire blight pathogen, or the surrounding area of ​​the host plant of the fire blight pathogen through various methods such as inoculation, spraying, immersion, drenching, or spraying.

[0039] Cyclo(-Met-Pro) according to one embodiment of the present invention may have the effect of inhibiting the growth of at least one of black rot bacteria, bacterial wilt bacteria, ulcer bacteria, or soft rot bacteria in addition to fire blight bacteria.

[0040] Accordingly, the present invention further provides at least one of a composition for inhibiting the growth of black rot pathogens, a composition for inhibiting the growth of bacterial wilt pathogens, a composition for inhibiting the growth of ulcer pathogens, or a composition for inhibiting the growth of soft rot pathogens, comprising cyclo(-Met-Pro) as an active ingredient.

[0041] The concentration of Cyclo(-Met-Pro) included as an active ingredient in the composition for inhibiting the growth of black wilt disease pathogens according to the present invention can be adjusted. For example, the concentration of Cyclo(-Met-Pro) may be 1 μg / mL to 10,000 μg / mL. In addition, for example, the concentration of Cyclo(-Met-Pro) may be 10 μg / mL to 7,000 μg / mL. In addition, for example, the concentration of Cyclo(-Met-Pro) may be 50 μg / mL to 5,000 μg / mL.

[0042] The concentration of Cyclo(-Met-Pro) included as an active ingredient in the composition for inhibiting the growth of bacterial wilt pathogens according to the present invention can be adjusted. For example, the concentration of Cyclo(-Met-Pro) may be 1 μg / mL to 10,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 10 μg / mL to 7,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 50 μg / mL to 5,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 2,000 μg / mL to 5,000 μg / mL.

[0043] The concentration of Cyclo(-Met-Pro) included as an active ingredient in the composition for inhibiting the growth of ulcer pathogens according to the present invention may be controlled. For example, the concentration of Cyclo(-Met-Pro) may be 1 μg / mL to 10,000 μg / mL. In addition, for example, the concentration of Cyclo(-Met-Pro) may be 10 μg / mL to 7,000 μg / mL. In addition, for example, the concentration of Cyclo(-Met-Pro) may be 1,000 μg / mL to 5,000 μg / mL. In addition, for example, the concentration of Cyclo(-Met-Pro) may be 2,000 μg / mL to 5,000 μg / mL.

[0044] The concentration of Cyclo(-Met-Pro) included as an active ingredient in the composition for inhibiting the growth of soft rot pathogens according to the present invention can be adjusted. For example, the concentration of Cyclo(-Met-Pro) may be 1 μg / mL to 10,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 10 μg / mL to 7,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 50 μg / mL to 5,000 μg / mL. Also, for example, the concentration of Cyclo(-Met-Pro) may be 2,000 μg / mL to 5,000 μg / mL.

[0045] The present invention will be described in more detail through examples. However, these examples are intended only to explain the invention in more detail and do not limit the scope of the invention.

[0047] Example 1: Growth inhibitory ability of fire blight control microorganisms against fire blight pathogens

[0048] In order to confirm the growth inhibitory ability of the fire blight control microorganisms in the present invention, the fire blight control microorganism Cerratia rubidea ( Serratia rubidaea ) The AO4-P6 strain was prepared. Cerratia rubidea ( Serratia rubidaea Since it has been confirmed through experiments that the cell suspension of the AO4-P6 strain inhibits fire blight pathogens, the present invention prepared an experiment to inhibit fire blight pathogens using the culture supernatant of the AO4-P6 strain in order to confirm whether substances produced by the AO4-P6 strain other than the cell suspension inhibit fire blight pathogens.

[0049] In one embodiment, Cerachia rubidea ( Serratia rubidaea The AO4-P6 strain was cultured in TSB (Tryptic Soy Broth) at 28°C for 24 hours, and the cells of the AO4-P6 strain were filtered to remove them. Cerratia rubidea ( Serratia rubidaea The culture supernatant of the AO4-P6 strain was inoculated at 1% in 1 / 5 TSB with Erwinia amylovora ( Erwinia amylovora )(OD 600 It was mixed at 10% (v / v) with (=0.01). In this case, a resazurin solution was mixed together as a cell growth indicator. Subsequently, color changes were observed after static culture at 28°C for 24 hours. In one example, Comparative Example 1-1 (Blank; Only 0.2 TSB), Comparative Example 1-2 (Negative control; Only E. amylovora Experimental Example 1-1 (Positive control 1; Streptomycin (µg / mL) 5 / 0.5 / 0.05), Experimental Example 1-2 (Positive control 2; Kanamycin (µg / mL) 5 / 0.5 / 0.05 / 0.005), and Experimental Example 1-3 (Supernatant of AO4-P6) were established.

[0050] Figure 1 is a photograph showing the results of an experiment on the growth inhibition ability of fire blight pathogens using the culture filtrate of the Cerratia rubidea AO4-P6 strain according to one embodiment of the present invention.

[0051] Referring to FIG. 1, the growth inhibitory ability of the culture supernatant of the Cerratia rubidea AO4-P6 strain according to one embodiment of the present invention against fire blight pathogens can be confirmed. Comparative Example 1-1 did not contain fire blight pathogens, so the color of the resazurin did not change, thereby confirming the color of the resazurin when the growth of the fire blight pathogens was inhibited. Comparative Example 1-2 contained fire blight pathogens and grew, causing the color of the resazurin to change, thereby confirming the color of the resazurin when the growth of the fire blight pathogens was not inhibited. Based on this, when examining Experimental Examples 1-1 to 1-3, the color of the resazurin in Experimental Examples 1-1 and 1-2 differed depending on the concentration of antibiotics (streptomycin, kanamycin), while the color of the resazurin in Experimental Example 1-3 did not change.

[0052] More specifically, in Experimental Example 1-1, when the streptomycin concentration was 5 μg / mL, the fire blight pathogen did not grow, and the color of the resazurin did not change as in Comparative Example 1-1. In Experimental Example 1-1, when the streptomycin concentration was 0.5 μg / mL or less (0.5 μg / mL, 0.05 μg / mL), the fire blight pathogen grew and the color of the resazurin changed, and the lower the concentration, the more similar the color was to that of Comparative Example 1-2.

[0053] In Experimental Example 1-2, when the kanamycin concentration was 0.05 μg / mL or higher (0.05 μg / mL, 0.5 μg / mL, 5 μg / mL), the fire blight pathogen could not grow, and the color of the resazurin did not change as in Comparative Example 1-1. In Experimental Example 1-2, when the kanamycin concentration was 0.005 μg / mL, the fire blight pathogen grew and the color of the resazurin changed.

[0054] In other words, in the experimental examples using antibiotics, when the antibiotic concentration was above the Minimum Inhibitory Concentration (MIC), the growth of the fire blight pathogen was inhibited, and no change in the color of resazurin was observed.

[0055] In Experimental Examples 1-3, when the culture filtrate of the AO4-P6 strain according to one embodiment of the present invention was treated, the fire blight pathogens could not grow, and the color of the resazurin did not change as in Comparative Example 1-1. Accordingly, it was confirmed that the culture filtrate of the AO4-P6 strain has growth inhibitory ability against fire blight pathogens.

[0057] Example 2: Isolation of Metabolites from AO4-P6 Strain and Search for Candidate Inhibitors Against Fire Blight Bacteria

[0058] In order to search for candidate substances to inhibit fire blight among the metabolites of the AO4-P6 strain, the metabolites of the AO4-P6 strain were separated and the growth inhibitory ability of each fraction of the fire blight strain was tested.

[0059] More specifically, the AO4-P6 strain was cultured on Tryptic Soy Agar (TSA) medium for 2 days, and after removing the cells, metabolites were extracted from the agar using methanol. The methanol extracts were concentrated under reduced pressure and fractionated using ODS or LH-20 MPLC; to test the growth inhibitory activity against fire blight pathogens, the fractions were tested in a 96-well system on Erwinia amylovora inoculated at 1% in 1 / 5 TSB ( Erwinia amylovora )(OD 600 =0.01) and 10% (v / v) were mixed. After static incubating the mixtures at 28°C for 24 hours, the fractions exhibiting growth inhibitory activity against fire blight pathogens were identified by measuring the OD values ​​of each mixture. In one example, Comparative Example 2-1 (Blank; Only 0.2 TSB), Comparative Example 2-2 (Negative control; MeOH, Only E. amylovoraExperimental Example 2-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 2-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Example 2-3 (Fractions) were established. Although Experimental Example 2-3 refers to multiple fractions, it was written as a single experimental example for brevity in this section, and fractions with significant differences in effect were described by their respective numbers.

[0060] FIG. 2 is a graph showing the degree of inhibition of fire wilt pathogen growth for each of the fractions separated according to one embodiment of the present invention.

[0061] Referring to FIG. 2, the inhibition of fire blight growth by each of the fractions isolated from the metabolites produced by the AO4-P6 strain according to one embodiment of the present invention (Experimental Examples 2-3) can be confirmed. More specifically, in the first isolation (ODS MPLC), growth inhibitory activity of fire blight was confirmed in sections 58 to 64 (Fraction A58 to Fraction A64), and subsequent isolation was carried out as it was determined that this section contained useful components as it was the 50% elution section during MPLC fractionation. In the second isolation (LH-20 MPLC), fire blight inhibitory activity was confirmed in sections 33 to 39 (Fraction 33 to Fraction 39).

[0062] Subsequently, chromatography was performed on some of the fractions (Fractions 30, 33, 36, 39, 41) according to the final fractionation to identify the peaks of each substance contained in some of the fractions.

[0063] FIG. 3 is a graph showing the chromatographic results of some of the fractions according to one embodiment of the present invention.

[0064] Referring to FIG. 3, the respective peaks formed by the substances included in the fractions according to one embodiment of the present invention can be observed. As previously discussed, it was confirmed that fraction 36 had the highest growth inhibitory activity against fire blight pathogens among fractions 30, 33, 36, 39, and 41. Accordingly, among the substances in fraction 36, substances with significantly higher peaks than those in other fractions were searched for, and as a result of the search, the peak formed at 11.7 min was selected as the candidate substance for inhibiting fire blight pathogens (Compound A).

[0065] Subsequently, structural analysis of the substance was performed using various spectra (Mass spectrum NMR spectrum) to determine the chemical structure of the active candidate substance. As a result of the structural analysis, the final structure of the fire blight inhibitory candidate substance of the present invention was identified as Cyclo(-Met-Pro), a cyclopeptide-based compound, and the detailed structure is as shown in Chemical Formula 1 below.

[0066]

[0068] Example 3: Inhibitory effect of candidate substance on fire blight bacteria

[0069] In the present invention, the growth inhibitory ability of the fire blight pathogen was tested on a candidate substance (Cyclo(-Met-Pro)) isolated through the previous example.

[0070] To test the growth inhibitory activity of the above substance against fire blight pathogens, Erwinia amylovora inoculated at 1% in 1 / 5 TSB with Cyclo(-Met-Pro) dilutions set at different concentrations (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) in a 96-well system Erwinia amylovora )(OD 600=0.01) and 10% (v / v) were mixed. The inhibitory effect on the growth of fire blight pathogens was confirmed by measuring the OD values ​​of each mixture after static incubation at 28 ℃ for 24 hours. In one example, Comparative Example 3-1 (Blank; Only 0.2 TSB), Comparative Example 3-2 (Negative control; MeOH, Only E. amylovora Experimental Example 3-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 3-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Examples 3-3 to 3-9 (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were established.

[0071] Figure 4 is a graph showing the degree of growth inhibition of fire blight pathogens by a candidate substance for inhibiting fire blight pathogens isolated according to one embodiment of the present invention.

[0072] Referring to FIG. 4, the inhibition of fire blight pathogen growth by a candidate substance for inhibiting fire blight pathogens according to a concentration set according to one embodiment of the present invention can be confirmed.

[0073] When the concentration of the fire blight inhibitor candidate substance (Cyclo(-Met-Pro)) was 100 μg / mL (Experimental Examples 3-3 and 3-4), the OD value decreased slightly compared to Comparative Example 3-1, and through this, it was confirmed that the fire blight inhibitor candidate substance (Cyclo(-Met-Pro)) with a concentration of 100 μg / mL or higher has an inhibitory effect on fire blight.

[0074] When the concentration of the candidate substance for inhibiting fire blight (Cyclo(-Met-Pro)) was 500 μg / mL to 5,000 μg / mL (Experimental Examples 3-5 to 3-9), the OD value decreased significantly compared to Comparative Example 3-1, and through this, it was confirmed that the candidate substance for inhibiting fire blight (Cyclo(-Met-Pro)) with a concentration of 500 μg / mL or higher has a significantly high inhibitory effect on fire blight. Accordingly, the candidate substance for inhibiting fire blight (Cyclo(-Met-Pro)) is described below as the fire blight inhibitor (Cyclo(-Met-Pro)).

[0076] Example 4: Inhibitory effect of a fire blight pathogen inhibitor on fire blight pathogens

[0077] In the present invention, the growth inhibitory ability against various plant pathogens was tested using a fire blight inhibitory substance (Cyclo(-Met-Pro)) isolated through the previous example.

[0078] To test the growth inhibitory activity of the above substance against various plant pathogens (black rot, bacterial wilt, canker, soft rot), Cyclo(-Met-Pro) dilutions set at different concentrations (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were inoculated at 1% in 1 / 5 TSB of Erwinia pyrifolia in a 96-well system Erwinia pyrifoliae ) (OD 600 =0.01), Ralstonia solanacearum( Ralstonia solanacearum )(OD 600 =0.01), Clavibacter miciganensis( Clavibacter michiganensis ) (OD 600 =0.01), or Pectobacterium carotoborum ( Pectobacterium carotovorum )(OD 600 =0.01) and 10% (v / v), respectively. After static incubating the mixtures at 28°C for 24 hours, the growth inhibitory effect on each plant pathogen was confirmed by measuring the OD value of each mixture.

[0079] In one embodiment, Comparative Example 4-1 (Blank; Only 0.2 TSB), Comparative Example 4-2 (Negative control; MeOH, Only E. pyrifoliae Experimental Example 4-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 4-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Examples 4-3 to 4-9 (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were established.

[0080] In addition, in one embodiment, Comparative Example 5-1 (Blank; Only 0.2 TSB), Comparative Example 5-2 (Negative control; MeOH, Only R. solanacearum Experimental Example 5-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 5-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Examples 5-3 to 5-9 (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were established.

[0081] In addition, in one embodiment, Comparative Example 6-1 (Blank; Only 0.2 TSB), Comparative Example 6-2 (Negative control; MeOH, Only C. michiganensis Experimental Example 6-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 6-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Examples 6-3 to 6-9 (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were established.

[0082] In addition, in one embodiment, Comparative Example 7-1 (Blank; Only 0.2 TSB), Comparative Example 7-2 (Negative control; MeOH, Only P. carotovorumExperimental Example 7-1 (Positive control 1; AO4-P6 supernatant), Experimental Example 7-2 (Positive control 2; Streptomycin or Kanamycin 5 μg / mL), and Experimental Examples 7-3 to 7-9 (100 μg / mL, 500 μg / mL, 1,000 μg / mL, 5,000 μg / mL) were established.

[0083] Figure 5 is a graph showing the degree of growth inhibition of black blight pathogens by a fire blight pathogen inhibitory substance isolated according to one embodiment of the present invention.

[0084] Referring to FIG. 5, the inhibition of the growth of black blight pathogens by a fire blight pathogen inhibitory substance according to a concentration set according to one embodiment of the present invention can be confirmed.

[0085] When the concentration of the fire blight inhibitory substance (Cyclo(-Met-Pro)) was 100 μg / mL or higher (Experimental Examples 4-3 to 4-9), the OD value decreased compared to Comparative Example 4-1, and through this, it was confirmed that the fire blight inhibitory substance (Cyclo(-Met-Pro)) at a concentration of 100 μg / mL or higher has an inhibitory effect on the black blight pathogen.

[0086] Figure 6 is a graph showing the degree of growth inhibition of bacterial blight pathogens by a fire blight pathogen inhibitory substance isolated according to one embodiment of the present invention.

[0087] Referring to FIG. 6, the inhibition of the growth of bacterial blight pathogens by a fire blight pathogen inhibitory substance according to a concentration set according to one embodiment of the present invention can be confirmed.

[0088] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 100 μg / mL to 1,000 μg / mL (Experimental Examples 5-3 and 5-7), the OD value decreased compared to Comparative Example 5-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) at a concentration of 100 μg / mL or higher has an inhibitory effect on bacterial wilt.

[0089] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 5,000 μg / mL (Experimental Examples 5-8 and 5-9), the OD value decreased significantly compared to Comparative Example 5-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) at a concentration of 2,000 μg / mL or higher has a significantly high inhibitory effect on the bacterial wilt disease.

[0090] Figure 7 is a graph showing the degree of inhibition of ulcer disease growth by a fire blight inhibitory substance isolated according to one embodiment of the present invention.

[0091] Referring to FIG. 7, the inhibition of ulcer disease growth by a fire blight inhibitory substance according to a concentration set according to one embodiment of the present invention can be confirmed.

[0092] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 1,000 μg / mL (Experimental Examples 6-7 and 6-8), the OD value decreased compared to Comparative Example 6-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) at a concentration of 1,000 μg / mL or higher has an inhibitory effect on ulcer pathogens.

[0093] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 5,000 μg / mL (Experimental Example 6-9), the OD value decreased significantly compared to Comparative Example 6-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) at a concentration of 5,000 μg / mL or higher has a significantly high inhibitory effect on ulcer pathogens.

[0094] Figure 8 is a graph showing the degree of growth inhibition of soft rot bacteria by a fire blight inhibitory substance isolated according to one embodiment of the present invention.

[0095] Referring to FIG. 8, the inhibition of soft rot growth by a fire blight inhibitory substance according to a concentration set according to one embodiment of the present invention can be confirmed.

[0096] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 100 μg / mL to 500 μg / mL (Experimental Examples 7-3 and 7-6), the OD value decreased compared to Comparative Example 7-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) at a concentration of 100 μg / mL or higher has an inhibitory effect on soft rot.

[0097] When the concentration of the fire blight inhibitor (Cyclo(-Met-Pro)) was 1,000 μg / mL to 5,000 μg / mL (Experimental Examples 7-7 to 7-9), the OD value decreased significantly compared to Comparative Example 7-1, and through this, it was confirmed that the fire blight inhibitor (Cyclo(-Met-Pro)) with a concentration of 1,000 μg / mL or higher has a significantly high inhibitory effect on soft rot.

[0098] Accordingly, it was confirmed that the fire blight fungus inhibitory substance (Cyclo(-Met-Pro)) according to one embodiment of the present invention has a growth inhibitory effect against fire blight fungus, as well as against black rot fungus, bacterial wilt fungus, ulcer fungus, and soft rot fungus. That is, the Cyclo(-Met-Pro) substance has a growth inhibitory effect against fire blight fungus, black rot fungus, bacterial wilt fungus, ulcer fungus, and soft rot fungus.

[0099] From the foregoing, those skilled in the art will understand that various changes and modifications are possible within the scope of the technical concept of the present invention. Accordingly, the technical scope of the present invention is not limited to the contents described in the detailed description of the specification, but must be determined by the claims.

Claims

Claim 1 A composition for inhibiting the growth of fire blight pathogens, comprising cyclo(-Met-Pro) as an active ingredient. Claim 2 A composition for inhibiting the growth of fire blight pathogens according to claim 1, wherein the cyclo(-Met-Pro) is included at a concentration of 100 μg / mL to 5,000 μg / mL. Claim 3 A composition for inhibiting the growth of fire blight pathogens according to claim 1, wherein the cyclo(-Met-Pro) has the effect of inhibiting the growth of at least one of fire blight pathogens, black rot pathogens, bacterial wilt pathogens, ulcer pathogens, or soft rot pathogens. Claim 4 The composition for inhibiting the growth of fire blight bacteria of claim 1 is Erwinia amylovora ( Erwinia amylovora A method for inhibiting the growth of fire blight pathogens, comprising the step of treating at least one of the surrounding area of ​​the Erwinia amylovora, the host plant of the Erwinia amylovora, or the surrounding area of ​​the host plant of the Erwinia amylovora. Claim 5 Erwinia Amylovora containing Cyclo(-Met-Pro) as an active ingredient ( Erwinia amylovora A composition for inhibiting fire blight, having a growth inhibitory effect on ). Claim 6 delete Claim 7 A composition for inhibiting the growth of black rot pathogens, comprising cyclo(-Met-Pro) as an active ingredient. Claim 8 A composition for inhibiting the growth of bacterial wilt pathogens, comprising cyclo(-Met-Pro) as an active ingredient. Claim 9 A composition for inhibiting the growth of ulcer pathogens, comprising Cyclo(-Met-Pro) as an active ingredient. Claim 10 delete

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