KR102389559B1 - Composition for controlling plant diseases comprising a compound having fungicidal activity derived from Pterocarya tonkinensis and method of controlling plant diseases using the same - Google Patents

Abstract

The present invention provides a composition for controlling plant diseases comprising at least one selected from the compounds represented by Formulas 1 to 8.
Pterocarya tonkinensis of the present invention　 tonkinensis ) The composition for controlling plant diseases containing a compound derived from it is harmless to the human body as a natural product, and is effective in controlling plant diseases without causing environmental pollution because it is biodegradable in nature and can be developed as an environmentally friendly biological pesticide. It can be usefully used in the production of high value-added organic products.

Description

Composition for controlling plant diseases comprising a compound having fungicidal activity derived from Pterocarya tonkinensis and a method for controlling plant diseases using the composition and method of controlling plant diseases using the same}

Pterocarya tonkinensis 　 tonkinensis ) It relates to a composition for controlling plant diseases comprising a bactericidal active compound isolated from an extract and a fraction thereof, and a method for controlling plant diseases using the composition.

Plant diseases are one of the biological hazard factors that hinder the healthy growth of plants, and affect agricultural production by impairing crop yield and quality. If a large-scale plant disease occurs in a large-scale cultivation area, not only will the crop yield be greatly reduced, resulting in a huge economic loss, but if there is a disruption in the main grain production, it will be a great threat in terms of food security. Serious social problems can arise.

In modern agriculture, chemical pesticides have been used as convenient means for controlling various plant diseases and minimizing losses by effectively managing plant diseases within an economic threshold range. However, as a result of increasing agricultural production by reliance on chemical pesticides for the past several decades, drug-resistant pathogens and pests appeared, concerns about human risk to livestock toxicity, and ecological disturbance due to residual toxicity occurred. There is a need to develop a new plant disease control means to reduce the use of and replace them.

A biopesticide is an eco-friendly plant disease control means used to control pathogens, pests and weeds using natural substances derived from nature, and plant extracts and microorganisms are used as main materials.

Plants biosynthesize various secondary metabolites such as alkaloids, terpenoids, flavonoids, glycosides, quinones, coumarins, and phenolic compounds in the body. Some of these substances act as antibiotics or phytoalexin to prevent invasion of pathogens or inhibit the feeding of pests. Rosemary oil (trade name: Milsana), rosemary oil (trade name: Sporan), and tea tree oil (trade name: Timorex) are commercially available as bio crop protection agents. A number of studies have been reported.

Pterocarya tonkinensis (Franch.) Dode belonging to the genus Pterocarya of the Juglandaceae family grows up to 30 m in height and is distributed in China, Vietnam, and Laos, and is distributed during the tropical monsoon. ) grows wild around rivers and streams in forests. Chinese Ginger ( Pterocarya ) stenoptera C. DC.) and subspecies ( Pterocarya ) are closely related ecologically and morphologically. stenoptera C. DC. var. tonkinensis Franch.). Locally, it is harvested and used as wood, and there is no report on its effectiveness in controlling plant diseases.

The present inventors as an environmentally friendly plant disease control means Pterocarya tonkinensis ( Pterocarya 　 tonkinensis ) plant extract was used.

Chinese Patent Publication 104961719 (2015. 10. 07.)

Natural Product Leters Volume 15(1), pp. 75-79 (2001)

One object of the present invention is to provide a composition for controlling plant diseases comprising at least one selected from the compounds represented by Formulas 1 to 8.

Another object of the present invention is to provide a method for controlling a plant disease comprising the step of treating the composition for controlling a plant disease on a plant, its seed or its habitat.

The present inventors were separated from the Pterocarya tonkinensis extract while investigating the control effect of various plant extracts on various plant diseases in order to develop an eco-friendly plant disease control agent. It has been found that the compounds used have an excellent plant disease control effect.

The terminology used in the present application is only used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly dictates otherwise. In the present application, terms such as “comprise” or “have” are intended to designate that a feature, step, structure, or a combination thereof described in the name exists, but one or more other features, steps, structure, or these It should be understood that it does not preclude the possibility of the existence or addition of combinations.

Unless defined otherwise, all terms used herein, including technical and scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present application. does not

In the present invention, “Cx-Cy” means having x to y carbon atoms.

In the present invention, “alkyl” refers to a linear (or straight-chain, linear) saturated hydrocarbon group or a branched (or branched, branched) saturated hydrocarbon group, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl , sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl and the like.

pterokarya tonkinensis ( Pterocarya tonkinensis ) from derived compound containing plant disease Controlling composition and method for preparing the same

In one aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising at least one selected from compounds represented by the following Chemical Formulas 1 to 8.

[Formula 1]

;

;

[Formula 2]

In the above formula,

R ₁ to R ₃ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl;

[Formula 3]

;

;

[Formula 4]

;

;

[Formula 5]

R ₄ to R ₆ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl;

[Formula 6]

;

;

[Formula 7]

In the above formula,

R ₇ and R ₈ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —OC ₁ -C ₆ alkyl,

R ₉ and R ₁₀ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —C ₁ -C ₆ alkyl-OH; and

[Formula 8]

.

.

In one aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising at least one selected from compounds represented by the following Chemical Formulas 1 to 8.

[Formula 1]

;

;

[Formula 2-1]

;

;

[Formula 2-2]

;

;

[Formula 2-3]

;

;

[Formula 3]

;

;

[Formula 4]

;

;

[Formula 5-1]

;

;

[Formula 5-2]

;

;

[Formula 6]

;

;

[Formula 7-1]

;

;

[Formula 7-2]

;

;

[Formula 7-3]

; 및

; and

[Formula 8]

.

.

In the present invention, the compound of Formula 1 is 1,7-(4"-hydroxy-3"-methoxy phenyl)-1-(4'-hydroxyphenyl)-4E-hepten-3-one (1 ,7-(4"-hydroxy-3"-methoxyphenyl)-1-(4'-hydroxyphenyl)-4E-hepten-3-one); The compound of Formula 2-1 is a pterocarine (pterocarine); The compound of Formula 2-2 is galeon; The compound of formula 2-3 is 3', 4"-epoxy-1-(4'-hydroxylphenyl)-7-(3"-methoxy-phenyl)-heptan-2-hydro-3-one (3' ,4"-epoxy-1-(4'-hydroxylphenyl)-7-(3"-methoxyl-phenyl)-heptane-2-hydro-3-one); The compound of formula 3 is (8R-cis)-6,7,8,8a-tetrahydro-5-hydroxy-8-(1-methylethyl)-2H-naphtho[1,8-bc]furan-2 -one ((8R-cis)-6,7,8,8a-tetrahydro-5-hydroxy-8-(1-methylethyl)-2H-naphtho[1,8-bc]furan-2-one); The compound of formula 4 is 9-hydroxymegastigm-7-en-3-one; The compound of formula 5-1 is saleenone (xylarenone); The compound of Formula 5-2 is 5,8-dihydroxy-4-methoxy-α-tetralone (5,8-dihydroxy-4-methoxy-α-tetralone); The compound of formula 6 is (E)-4-hydroxycinnamic acid methyl ester ((E)-4-hydroxycinnamic acid methyl ester); The compound of Formula 7-1 is an asiatic acid; The compound of Formula 7-2 is arjunolic acid; The compound of Formula 7-3 is maslinic acid; The compound of Formula 8 is called loranthol.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (1).

[Formula 1]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (2).

[Formula 2]

In the above formula,

R ₁ to R ₃ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl.

Preferably, R ₁ to R ₃ are each independently a hydrogen atom, -OH or -OC ₁ -C ₆ alkyl, more preferably R ₁ and R ₂ are each independently, -OH or -OC ₁ -C ₆ alkyl, and R ₃ is a hydrogen atom or -OH.

The compound represented by Formula 2 may most preferably be any one of compounds represented by the following Formulas 2-1 to 2-3, and the composition of the present invention is represented by the following Formulas 2-1 to 2-3 It may include one or more of the compounds used.

[Formula 2-1]

;

;

[Formula 2-2]

; 및

; and

[Formula 2-3]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (3).

[Formula 3]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (4).

[Formula 4]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (5).

[Formula 5]

R ₄ to R ₆ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl.

Preferably, R ₄ to R ₆ may each independently be a hydrogen atom, -OH or -OC ₁ -C ₆ alkyl.

Most preferably, the compound represented by Formula 5 may be any one of compounds represented by Formulas 5-1 and 5-2, and the composition of the present invention is a compound represented by Formulas 5-1 and 5-2 It may include one or more of

[Formula 5-1]

; 및

; and

[Formula 5-2]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (6).

[Formula 6]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (7).

[Formula 7]

In the above formula,

R ₇ and R ₈ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —OC ₁ -C ₆ alkyl,

R ₉ and R ₁₀ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —C ₁ -C ₆ alkyl-OH.

Preferably, R ₇ and R ₈ are each independently a hydrogen atom or C ₁ -C ₆ alkyl, R ₉ and R ₁₀ are each independently C ₁ -C ₆ alkyl or —C ₁ -C ₆ alkyl-OH am.

More preferably, the compound represented by Chemical Formula 7 may be a compound represented by the following Chemical Formula 7a or 7b.

[Formula 7a]

[Formula 7b]

In Formulas 7a and 7b, the descriptions of R ₇ , R ₈ , R ₉ and R ₁₀ are the same as described above.

Most preferably, the compound represented by Formula 7 may be any one of compounds represented by Formulas 7-1 to 7-3, and the composition of the present invention is a compound represented by Formulas 7-1 to 7-3 It may include one or more of

[Formula 7-1]

;

;

[Formula 7-2]

; 및

; and

[Formula 7-3]

.

.

In another aspect of the present invention, the present invention provides a composition for controlling plant diseases comprising a compound represented by the following formula (8).

[Formula 8]

.

.

In the present invention, the compound may have a bactericidal activity, and the sterilization may mean physically or chemically destroying all types of microorganisms such as molds, viruses, and bacteria, and may also be sterilization.

In the present specification, the term "control" is intended to be used in the meaning of preventing and avoiding diseases or pests, as well as removing and killing them. However, in the control of plant diseases, the control effect by prophylactic treatment is the main control mechanism, so the investigation of the control effect of the compounds represented by Chemical Formulas 1 to 8 on plant diseases was tested as a preventive treatment.

In the present invention, all of the compounds that can be included in the composition are derived from natural products, are harmless to the human body, and do not cause environmental pollution, while Alternaria Brassica ( Alternaria ) brassicicola ; Brassica crops black-spotted fungus), Botrytis cinerea (Grey fungus), Colletotrichum coccodes ; Chili anthrax), Fusarium oxysporum (wilt), Magnaporthe oryzae ; Rice Blast Bacteria), Puccinia triticina (Wheat Red Rust Bacteria) and Phytophthora Phytophthora infestans ; Potato / tomato late blight), such as phytopathogenic fungi or Abene subsp. Cattray ( Acidovorax) avenae subsp. cattleyae ; Phalaenopsis orchid bacterial brown spot bacterium), Agrobacterium tumefaciens ( Agrobacterium tumefaciens ; fruit tree root-knot), Clavibacter misiganensis subsp. Clavibacter michiganensis subsp. michiganensis ; It exhibits an excellent control effect on plant diseases, which are important in crop production, caused by phytopathogenic bacteria such as pepper ulcer disease), and factors causing plant diseases are not limited thereto.

In the present invention, the compounds represented by Formulas 1 to 8 are Pterocarya tonkinensis ( Pterocarya )　 tonkinensis may be derived from, preferably Pterocarya tonkinensis ( Pterocarya 　 tonkinensis ) may be isolated from an extract or a fraction thereof.

In the present invention, the extract is water, C ₁ to C ₄ lower alcohols (eg, methanol, ethanol, propanol, isopropanol, butanol), hexane, chloroform, methylene chloride, ethyl acetate, acetone, acetonitrile and The extraction may be performed using a combination thereof as a solvent, but the extraction solvent is not limited thereto.

In the present invention, the fraction of the extract may be fractionated using hexane, chloroform, ethyl acetate, methylene chloride, methanol, ethanol, butanol, water, and combinations thereof as a solvent, but the fractionation solvent is not limited thereto .

In the present invention, the extract may be a leaf, stem or root extract of Pterocarya tonkinensis , and the site for extracting the Pterocarya tonkinensis extract is not limited thereto.

In the present invention, by using the fraction of the extract, the compounds were isolated and identified, and 13 new compounds having an effect of controlling plant diseases were obtained.

In the present invention, the compound may be included in the composition for controlling plant diseases at a concentration of 100 to 5000 μg/ml. The concentration represents the concentration contained in the composition when treating plants, seeds or habitats thereof, in which plant diseases have occurred, and when the concentration is less than 100 μg / ml, the concentration of the compound is too low, the control effect of plant diseases is There is a problem of falling, and when the concentration is more than 5000 μg/ml, the concentration of the compound is too high than necessary, which is uneconomical and may have a negative impact on the environment. However, the amount of the composition for controlling plant diseases is not limited to the above concentration range, and may be appropriately adjusted in consideration of the type of plant pathogen or phytopathogenic bacteria, the degree of occurrence, the environment, and the like.

The composition of the present invention may be a mixture further comprising an inert carrier, and a surfactant and It is prepared by adding other auxiliaries as needed. The above-mentioned composition for controlling plant diseases by itself or by adding other inactive ingredients, the present invention can also be used as a seed treatment agent.

In the present invention, examples of the liquid carrier that can be used in the formulation include water; alcohols such as methanol and ethanol; ketones such as acetone and methyl ethyl ketone; aromatic hydrocarbons such as benzene, toluene, xylene, ethylbenzene and methylnaphthalene; aliphatic hydrocarbons such as hexane, cyclohexane, kerosine and light oil; esters such as ethyl acetate and butyl acetate; nitriles such as acetonitrile and isobutyrnitrile; ethers such as diisopropylether and dioxane; acid amides such as N,N-dimethyl formamide and N,N-dimethylacetamide; halogenated hydrocarbons such as dichloromethane, trichloroethane and carbon tetrachloride; dimethyl sulfoxide; and vegetable oils such as soybean oil and cottonseed oil.

Further, examples of solid carriers that can be used in the formulation include fine powders or granules such as minerals such as kaolin clay, attapulgite clay, bentonite, montmorillonite, acid white clay, pyrophyllite, talc, diatomaceous earth and talcite; natural organic substances such as corn leaf stalk powder and walnut hull powder; synthetic organic materials such as urea; salts such as calcium carbonate and ammonium sulfate; synthetic inorganic materials such as synthetic hydrated silicon oxide; As liquid carriers, aromatic hydrocarbons such as xylene, alkylbenzene and methylnaphthalene; alcohols such as 2-propanol, ethylene glycol, propylene glycol and ethylene glycol monoethyl ether; ketones such as acetone, cyclohexanone and isophorone; vegetable oils such as soybean oil and cottonseed oil; petroleum aliphatic hydrocarbons, esters, dimethylsulfoxide, acetonitrile and water.

Further, examples of surfactants include anionic surfactants such as alkyl sulfate ester salts, alkylaryl sulfonate salts, dialkylsulfosuccinate salts, polyoxyethylene alkylaryl ether phosphate ester salts, lignosulfonate salts and naphthalene sulfo nate formaldehyde polycondensate; and nonionic surfactants such as polyoxyethylene alkyl aryl ethers, polyoxyethylene alkylpolyoxypropylene block copolymers and sorbitan fatty acid esters and cationic surfactants such as alkyltrimethylammonium salts.

Examples of other formulation aids include water-soluble polymers such as polyvinyl alcohol and polyvinylpyrrolidone, polysaccharides such as gum arabic, alginic acid and salts thereof, CMC (carboxymethyl-cellulose), xanthan gum, inorganic substances such as aluminum magnesium silicate and alumina sol ( alumina sol), preservatives, colorants and stabilizers such as PAP (acid phosphate isopropyl) and BHT (butylhydroxyltoluene).

In one embodiment of the present invention, it was confirmed through Examples that the compound isolated from the Pterocaya tonkinensis extract and its fractions had a control effect on plant pathogens, and from this, the compound isolated from the extract or fraction It can be seen that the composition for controlling plant diseases comprising a plant disease control effect.

The Pterokarya tonkinensis extract or fraction may be prepared by a manufacturing method comprising the following steps, and the identification of the compound therefrom may undergo the following additional steps, but is not limited thereto:

The method for preparing the Pterokarya tonkinensis fraction may exemplarily pass through the following steps:

1) extracting by adding an extraction solvent to Pterokarya tonkinensis;

2) filtering the extract of step 1);

3) Concentrating the filtered extract of step 2) under reduced pressure and drying it to prepare an extract of Pterocarya tonkinensis; and

4) preparing a Pterokarya tonkinensis fraction by additionally extracting the Pterokarya tonkinensis extract of step 3) with an organic solvent.

In the above method, the Pterokarya tonkinensis of step 1) may be used without limitation, such as cultivated or commercially available ones. The Pterokarya tonkinensis may have leaves, stems or roots, but is not limited thereto.

The extraction solvent of step 1) may be any one selected from the group consisting of water, C ₁ to C ₄ lower alcohol, hexane, chloroform, methylene chloride, ethyl acetate, acetone and acetonitrile, or a mixed solution thereof, However, the present invention is not limited thereto. The lower alcohol may be methanol, ethanol, propanol, isopropanol or butanol.

As the extraction method, shaking extraction, Soxhlet extraction, or reflux extraction may be used, but the present invention is not limited thereto. The extraction solvent can be extracted by adding 1 to 10 times the amount of dried Pterokarya tonkinensis, and can be extracted by adding 2 to 3 times. The extraction temperature may be in the range of 20°C to 100°C, and may be in the range of 20°C to 40°C, and may be room temperature, but is not limited thereto. In addition, the extraction time may be 10 to 48 hours, may be 15 to 30 hours, may be 24 hours, but is not limited thereto. Furthermore, the number of times of extraction may be 1 to 5 times, may be repeated extraction 3 to 4 times, and may be 3 times, but is not limited thereto.

In the method, the vacuum concentration in step 3) may be performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

In the method, the organic solvent of step 4) may be any one selected from the group consisting of hexane, chloroform, ethyl acetate, methylene chloride, methanol, ethanol, butanol and water or a mixture thereof, but is not limited thereto. The fraction may be any one of a solvent fraction obtained by suspending the Pterokarya tonkinensis extract in water and fractionating it with hexane, ethyl acetate, butanol or water, and may be an ethyl acetate fraction or a butanol fraction, but is limited thereto it's not going to be The fraction may be obtained by repeating the fractionation process 1 to 5 times or 3 times from the Pterokarya tonkinensis extract, and may be concentrated under reduced pressure after fractionation, but is not limited thereto.

The separation and identification method of the compound from the Pterokarya tonkinensis fraction can be exemplarily performed in the following manner:

First, the method for preparing the Pterokarya tonkinensis fraction may be exemplarily performed in the following steps:

1) extracting by adding an extraction solvent to Pterokarya tonkinensis;

2) filtering the extract of step 1);

3) Concentrating the filtered extract of step 2) under reduced pressure and drying it to prepare an extract of Pterocarya tonkinensis;

4) additionally extracting the Pterocarya tonkinensis extract of step 3) with an organic solvent to prepare a Pterokarya tonkinensis fraction; and

5) separating the fraction of step 4) by at least one selected from the group consisting of column chromatography, medium pressure liquid chromatography, and high pressure liquid chromatography to obtain any one or more of the compounds represented by Formulas 1 to 8 .

In the above method, step 5) is a step of further separating and purifying the fraction using a conventional chromatography method to provide a single compound. Specifically, a fraction is prepared by eluting under an appropriate elution solvent condition through silica gel column chromatography, medium pressure liquid chromatography, etc., and it is further separated by medium pressure liquid chromatography, high pressure liquid chromatography, etc. under concentration gradient conditions, as mentioned above One compound of Formulas 1 to 8 can be obtained.

In this regard, a schematic diagram of a specific manufacturing process is shown in FIG. 1 .

That is, in the present invention, a solvent fraction is prepared from the Pterokarya tonkinensis extract, and the fractions are fractionated by liquid chromatography to separate the compounds represented by Chemical Formulas 1 to 8.

The liquid chromatography technique refers to a chromatography technique in which a mobile phase is a liquid, and is performed in a column filled with a stationary phase or a plane to which a stationary phase is attached. The pterokarya tonkinensis fraction is not limited as long as it is a solvent fraction, but may be a hexane fraction, a chloroform fraction, or an ethyl acetate fraction, but is not limited thereto. As the mobile phase, organic solvents such as water, hexane, methanol, ethanol, acetonitrile, acetone, chloroform, dichloromethane, and ethyl acetate may be used alone or in combination, and as the stationary phase, silica gel, Diaion HP-20 , RP-18 or Sephadex LH-20 may be used, but is not limited thereto.

The compounds represented by Formulas 1 to 8 are isolated from a fraction thereof of the Pterocaya tonkinensis extract, and may have bactericidal activity.

The compound represented by any one of Formulas 1 to 8 of the present invention includes not only salts thereof, but also solvates, optical isomers, hydrates, and the like, which can be prepared therefrom by a conventional method.

In the present invention, as the “salt” of the compound represented by any one of Formulas 1 to 8, an acid addition salt formed by a free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc., and organic acids such as acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride. , acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, Fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, methoxybenzoate, Phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, βhydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

In addition, the acid addition salt can be prepared by a conventional method, for example, at least one of the compounds represented by Formulas 1 to 8 is dissolved in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. It can be prepared by filtering and drying the precipitate generated by adding an organic or inorganic acid, or by distilling the solvent and excess acid under reduced pressure and drying it to crystallize it in an organic solvent.

Also, bases can be used to make metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

In the present invention, “isomer” includes “stereoisomer” or “enantiomer”, but stereoisomer may include each or mixture of diastereomers, and optical isomers include Each of the enantiomers as well as mixtures of enantiomers and racemates are included.

The “hydrate” of the present invention refers to a compound represented by any one of Formulas 1 to 8 to which water is non-covalently bound by intermolecular force, and may include a stoichiometric or non-stoichiometric amount of water. Specifically, the hydrate may contain water in a ratio of about 0.25 to about 10 moles based on 1 mole of the active ingredient, and more specifically, about 0.5 moles, about 1.5 moles, about 2 moles, about 2.5 moles, about It may include 3 moles, etc., but is not limited thereto.

The “solvate” of the present invention is one in which the compound represented by any one of Formulas 1 to 8 is bound to a solvent other than water by an intermolecular force, and may include a stoichiometric or non-stoichiometric amount of water. . Specifically, the hydrate may contain water in a ratio of about 0.25 to about 10 moles based on 1 mole of the active ingredient, and more specifically, about 0.5 moles, about 1.5 moles, about 2 moles, about 2.5 moles, about It may include 3 moles, about 5 moles, etc., but is not limited thereto.

pterokarya tonkinensis ( Pterocarya tonkinensis ) from derived compound containing plant disease using the control composition plant disease control method

In another aspect of the present invention, a plant disease control method comprising; treating a plant, its seed or its habitat with a plant disease control composition comprising at least one of the compounds represented by Formulas 1 to 8 provides

The treatment may be indirect spraying of spraying the composition directly on the plant, spraying on the soil in which the plant is growing, or spraying on the medium for culturing the plant.

The control method of the present invention includes treatment of stems and leaves of plants, treatment of places where plants grow (eg soil), treatment of seeds such as seed sterilization/seed coating and treatment of roots.

As treatment of stems and leaves with the control method of the present invention, application on the surface of plants, such as, for example, spraying on stems and leaves, in particular can be included. As the treatment of the soil with the control method of the present invention, for example, spraying onto the soil, mixing with the soil, spraying the liquid treatment agent into the soil (irrigation of the liquid treatment agent, injection into the soil, dripping of the liquid treatment agent) can be included. Examples of places to be treated include planting holes, furrows, around replanting, around planting furrows, the entire surface of the growth site, the area between soil and plants, the area between roots, and the area under the stem of a plant. , main furrow, growing soil, nail bed. Includes a seedling box, a seedling tray, and a seedbed. The treatment may be performed before spraying, during spraying, immediately after spraying, during the simulated cultivation period, before cultivation settlement, at the time of cultivation settlement, and during the growth period after cultivation settlement. In the above-mentioned soil treatment, the active ingredient may be simultaneously applied to the plant, or a solid fertilizer such as a paste fertilizer containing the active ingredient may be applied to the soil. The active ingredient may be mixed in an irrigation liquid, for example injected into an irrigation plant (irrigation tube, irrigation pipe, sprinkler, etc.), mixed in a liquid overflowing between furrows, or mixed into a water culture medium. can Alternatively, the irrigation liquid and active ingredient may be mixed beforehand and used for treatment by suitable irrigation methods, including, for example, the irrigation methods mentioned above and other methods such as spraying and flooding.

The volatilization treatment method as the control method of the present invention is, for example, the soil in which the plant is cultured with the composition for controlling plant diseases of the present invention, a hydroponic medium for culturing plants, and the volatilization of the sprayed composition. It is a method of protecting the plant from pests and diseases through the use of a method, and in addition, the composition can be placed around the plant to expose the plant to the composition in the volatilized gaseous state.

The seed treatment method in the control method of the present invention is, for example, a method of treating seeds so as to be protected from pests with the composition for controlling plant diseases of the present invention, and a specific example thereof is atomization of a suspension of the composition for controlling plant diseases of the present invention. and a spray treatment method of spraying on the seed surface; a spraying treatment method of applying the wettable powder, emulsion, glidant, etc. of the composition for controlling plant diseases of the present invention by itself or by adding a small amount of water to the seed surface; Impregnation treatment method of impregnating the seeds in the solution of the composition for controlling plant diseases of the present invention for a specific period; including film coating treatment and pellet coating treatment.

When a plant or soil for plant growth is treated with the compound according to the present invention, the treatment amount may vary depending on the type of plant to be treated, the type and frequency of occurrence of the pest to be controlled, the formulation form, the treatment period, climatic conditions, and the like.

Emulsions, wettable powders, glidants, etc. are usually diluted with water and then applied for treatment. In this case, the concentration of the active ingredient is usually in the range of 0.0001 to 3% by weight, preferably 0.0005 to 1% by weight. Powders, granules, and the like are usually used for treatment without dilution.

The control method of the present invention can be used in cultivated land or non-arable land such as paddy fields.

Pterocarya tonkinensis 　 tonkinensis ) The composition for controlling plant diseases containing a compound derived from it is harmless to the human body as a natural product, and is effective in controlling plant diseases without causing environmental pollution because it is biodegradable in nature and can be developed as an environmentally friendly biological pesticide. It can be usefully used in the production of high value-added organic products.

1 is Pterocarya tonkinensis ( Pterocarya 　 tonkinensis ) is a process diagram for separating compounds having antibacterial activity from the fraction of the extract.

Hereinafter, it will be described in detail by way of Examples.

However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

< Example 1> pterokarya tonkinensis containing extracts plant disease of the control composition plant disease Control effectiveness evaluation

Step 1: pterokarya tonkinensis Preparation of extracts

In order to prepare a composition for controlling plant diseases including the Pterokarya tonkinensis extract, first, the Pterokarya tonkinensis extract was prepared. Specifically, 3.8 kg of a leaf/stem sample of Pterocarya tonkinensis was cut into small pieces, methanol was added, extracted at room temperature for 24 hours, filtered through a filter paper, and the filtrate was concentrated under reduced pressure to 190 g of methanol extract of Pterocarya tonkinensis. got

Step 2: pterokarya tonkinensis containing extracts plant disease Preparation of the control composition

For the evaluation experiment of the plant disease control effect of the composition for controlling plant diseases containing the extract of Pterokarya tonkinensis, 2 ml of the extract sample of step 1 dissolved in methanol at a concentration of 60 mg/ml was diluted with 38 ml of distilled water, and spreader Tween 20 ( Tween 20) was added to a concentration of 0.025% (v/v) to prepare 40ml of a composition solution for controlling plant diseases containing a Pterocarya tonkinensis extract at a concentration of 3000μg/ml.

Step 3: pterokarya tonkinensis containing extracts plant disease of the control composition plant disease Control activity investigation

Rice blast disease of the control composition prepared in step 2 (pathogen: Magnaporthe oryzae ), tomato gray mold ( Pathogen: Botrytis cinerea ), Tomato late blight ( Pathogen: Phytophthora ) infestans ), wheat red rust (pathogen: Puccinia ) triticina ) and red pepper anthracnose (pathogen: Colletotrichum coccodes ) were tested for disease control activity against five plant diseases in greenhouse conditions as follows. In this case, as a control, distilled water containing 5% (v/v) methanol and 0.0025% (v/v) Tween 20 was used. The experimental results are shown in Table 1.

Specifically, three ports were used for each plant pathogen. After spraying the control composition solution prepared in step 2 on the leaf surface (foliage), it was air-dried for 24 hours, and then each plant pathogen was inoculated. For the rice, tomato, barley, pepper and wheat plants used in the experiment, fill a plastic pot with a diameter of 4.5 cm with about 70% of water supply or horticultural top soil, and then sow the seeds in a greenhouse at 25±5℃ for 1 to 4 weeks. cultivated.

Rice blast is the causative bacterium, Magnaporthe , in seedlings of the 3-4 leaf stage. oryzae KACC 46552)) was spray inoculated with a spore suspension (5×10 ⁵ spores/ml), treated for one day in a humid room at 25°C, and cultivated for 4 days in a constant temperature and humidity room (25°C, relative humidity 80%). was induced.

Tomato blight is caused by the causative bacterium Phytophthora ( Phytophthora ) in tomato seedlings in the 3-4 leaf stage. infestans KACC 48738), the sporangia suspension exposed from the sporangia (5×10 ⁴ sporangia/ml) was spray-inoculated, treated for 2 days in a humid room at 20°C, and cultured for 1 day in a constant temperature and humidity room at 20°C to induce disease.

Tomato gray mold disease is caused by Botrytis cineria , the causative agent of gray mold disease, on seedlings in the 3-4 leaf stage of tomatoes. cinerea KACC 48736) was inoculated with a spore suspension (5×10 ⁵ spores/ml), and then cultured for 3 days in a humid room at 20° C. to induce disease.

Wheat red rust is caused by Puccinia triticina, the causative bacterium of rust known as live parasite, on first-leaf seedlings. triticina (Korea Research Institute of Chemical Technology) in 250 μg/ml Tween 20 solution, 0.67 g of rust spores per liter, shake suspended, spray inoculated, and moistened in a humid room at 20 ° C for one day, and then in a constant temperature and humidity room at 20 ° C. It was transferred and cultured for 6 days to induce disease.

Capsicum anthrax is the causative agent of chilli anthrax, Colletotrichum coccodes KACC 48737) of spore suspension (4×10 ⁵ spores/ml) was spray inoculated, and after 2 days of moist treatment in a humid room at 25°C, it was moved to a constant temperature and humidity room (25°C, relative humidity 80%) and cultured for 1 day. was induced.

Tomato gray mold, tomato late blight and red pepper anthrax were investigated 3 days after inoculation, rice blast 5 days after inoculation, and wheat red rust and barley powdery mildew 7 days after inoculation, the lesion area (%) formed on the leaves was investigated.

The control value (%) was calculated according to Equation 1 below using the lesion area ratio (%) obtained from the above.

[Equation 1]

Control value (%) = (1-Less area ratio of treated group / lesion area ratio of untreated group) X 100

sample Concentration (μg/ml) Control cost (%) RCB TGM TLB WLR PAN Pterokarya tonkinensis extract 3000 85 21 93 67 92

(RCB: rice blast disease, TGM: tomato gray mold disease, TLB: tomato late blight, WLR: wheat red rust, PAN: red pepper anthrax)

As shown in Table 1, when the extract obtained with methanol from the leaves and stems of Pterokarya tonkinensis was treated at a concentration of 3000 μg/ml, the control effect on plant diseases was exhibited. In particular, it showed an excellent control effect of more than 92% against tomato late blight and red pepper anthrax, and showed a high control effect of 85% against rice blast disease. In the past, the plant disease control effect of the extract of Pterokarya tonkinensis has not been reported.

< Example 2> pterokarya tonkinensis of extract solvent fraction containing plant disease of the control composition plant disease Control effectiveness evaluation

Step 1: pterokarya tonkinensis the solvent of the extract fraction of Produce

In order to prepare a composition for controlling plant diseases comprising a solvent fraction of the Pterocarya tonkinensis extract, a methanol extract of Pterokarya tonkinensis was prepared by performing the same method as in Step 1 of Example 1, Solvent fractions were prepared using the methanol extract.

Specifically, 190 g of the methanol extract of Pterocarya tonkinensis leaves and stems was dissolved in 2L of distilled water and extracted twice with the same amount of hexane to obtain a hexane layer and an aqueous solution layer. The aqueous layer was again extracted twice with equal amounts of chloroform and ethyl acetate, respectively, to obtain a chloroform layer, an ethyl acetate layer and an aqueous solution layer. Then, these were concentrated under reduced pressure to obtain a dried hexane fraction (36 g), a chloroform fraction (33 g), and an ethyl acetate fraction (14 g).

Step 2: pterokarya tonkinensis the solvent of the extract fraction containing plant disease Preparation of the control composition

For the evaluation experiment of the plant disease control effect of the composition for controlling plant diseases containing the solvent fraction of the Pterokarya tonkinensis extract, 2 ml of each sample dissolved in methanol at a concentration of 40 mg/ml was diluted with 38 ml of distilled water, and the surfactant Tween 20 was added to a concentration of 0.025% (v/v), and 40 ml each of a composition solution for controlling plant diseases containing a solvent fraction of Pterocarya tonkinensis extract at a concentration of 2000 μg/ml was prepared.

Step 3: pterokarya tonkinensis containing extract plant disease of the control composition plant disease Check control activity

Rice blast disease of the control composition prepared in step 2 (pathogen: Magnaporthe oryzae ), tomato gray mold ( Pathogen: Botrytis cinerea ), Tomato late blight ( Pathogen: Phytophthora ) infestans ), wheat red rust (causative bacterium: Puccinia ) triticina ) and red pepper anthrax (pathogen: Colletotrichum coccodes ) were tested in the same manner as in step 3 of Example 1 in greenhouse conditions for disease control activity against plant diseases. In this case, as a control, distilled water containing 5% (v/v) methanol and 0.025% (v/v) Tween 20 was used. The experimental results are shown in Table 2.

process density
(μg/ml) Disease Control Value (%) RCB TGM TLB WLR PAN hexane fraction 2000 79 91 98 83 94 chloroform fraction 2000 79 36 88 83 92 ethyl acetate fraction 2000 79 21 94 60 70

(RCB: rice blast disease, TGM: tomato gray mold disease, TLB: tomato late blight, WLR: wheat red rust, PAN: red pepper anthrax)

As shown in Table 2, the hexane fraction showed an excellent control effect of 91% to 98% against tomato gray mold, tomato late blight, and pepper anthrax at a concentration of 2000 μg/ml, and 79% against rice blast and wheat red rust. It showed a control value of to 83%.

At the same concentration, the chloroform fraction showed an excellent control effect of 92% against red pepper anthrax, and showed a control value of 79% to 88% against rice blast, tomato late blight, and wheat red rust.

At the same concentration, the ethyl acetate fraction showed a high control value of 94% against tomato late blight, and showed a control value of 60% to 79% against rice blast, wheat red rust, and red pepper anthrax.

From the above results, it was confirmed that the antibacterial active compound of the Pterokarya tonkinensis extract was a non-polar material, not a water-soluble material. The hexane fraction, chloroform fraction and ethyl acetate fraction showed the highest overall control activity against several plant diseases, and were used for separation and purification of bactericidal active ingredients.

< Example 3> plant disease Isolation of compounds with control activity

Silica gel (Kiesel gel 60; 70-230mesh; Merck) column chromatography, MCI gel CHP 20/P120 (Misubishi Chemical Industries) column chromatography, Biotage Isolera One Separation was performed using a combination of a mid-pressure liquid chromatography (MPLC) system and a Shimadzu Prominence high pressure liquid chromatography (HPLC) system.

Specifically, compound 2-1, compound 3, compound 4, compound 7-3, and compound 2-1, compound 3, compound 4, compound 7-3 and Compound 8 was isolated.

In more detail, the hexane fraction (36 g) was adsorbed on silica gel, and then a hexane-ethyl acetate solution mixed in a volume ratio of 0:100 to 100:0 was sequentially eluted to elute 10 fractions (H1, H2, H3, H4, H5, H6, H7, H8, H9 and H10). After adding the active fraction H8 (886.0 mg) to a Biotage SNAP KP-Sil (50g) column, dichloromethane-ethyl acetate solution was eluted under a gradient of 0%-50% (v/v) to 7 fractions (H8A, H8B, H8C, H8D, H8E, H8F and H8G). After the active fraction H8D (455.9 mg) was added to a Biotage SNAP Ultra (30 g) column, an aqueous methanol solution was eluted under a gradient condition of 60%-100% (v/v), and compound 2-1 (24.3 mg), compound 3 (22.8) mg), and compound 4 (6.5 mg) was isolated. After adding the active fraction H9 (2.9 g) to a Biotage SNAP KP-Sil (50 g) column, dichloromethane-acetone solution was eluted with a gradient of 0%-50% (v/v) to 10 fractions (H9A, H9B). , H9C, H9D, H9E, H9F, H9G, H9H, H9I and H9J). After the active fraction H9I (189.5 mg) was added to a Biotage SNAP Ultra (30 g) column, an aqueous methanol solution was eluted under a gradient condition of 70%-100% (v/v), and compound 7-3 (59.7 mg) and compound 8 (9.3 mg) was isolated.

In order to separate the bactericidal active compound from the chloroform fraction (33 g) obtained in step 1 of Example 2, compound 1, compound 2-1, compound 2-2, compound 2-3 by the same process as shown in FIG. , compound 5-1, compound 5-2 and compound 6 were isolated.

In more detail, the chloroform fraction (33 g) was adsorbed on silica gel, followed by 100:0, 98:2, 95:5, 92:8, 88:12, 80:20, 60:40 and 25:75 The dichloromethane-ethyl acetate solution mixed in a volume ratio was sequentially eluted and divided into 8 fractions (C1, C2, C3, C4, C5, C6, C7 and C8). After adding the active fraction C3 (1.5g) to a Biotage SNAP Ultra (30g) column, eluting with an aqueous methanol solution under a gradient condition of 40%-100% (v/v) 9 fractions (C31, C32, C33, C34, C35) , C36, C37, C38 and C39), and the active fraction C32 was named compound 2-2 (67.8 mg). The active fraction C31 (179.9 mg) was loaded onto a YMC-Pack ODS-A column (20×250 mm, 5 μm) connected to a Shimadzu Prominence HPLC system, and eluted with a 67% aqueous methanol solution at a flow rate of 5 ml/min to obtain compound 5-1 (54.7). mg), compound 5-2 (28.7 mg) and compound 6 (4.9 mg) were isolated. The active fraction C4 (3.1 g) was adsorbed on MCI gel CHP 20/P120 and then eluted with an acetonitrile aqueous solution under a concentration gradient of 20%-100% (v/v) to obtain 16 fractions (C41, C42, C43, C44). , C45, C46, C47, C48, C49, C410, C411, C412, C413, C414, C415 and C416), and the active fraction C49 was designated as compound 2-3 (16.6 mg). The active fraction C410 (74.5 mg) was loaded on a YMC-Pack ODS-A column (20 × 250 mm, 5 μm) connected to a Shimadzu Prominence HPLC system, and 62% aqueous methanol solution (containing 0.1% formic acid) was eluted at a flow rate of 5 ml/min. Compound 1 (20.0 mg) and compound 2-1 (40.0 mg) were isolated.

In order to separate the bactericidal active compound from the ethyl acetate fraction (14 g) obtained in step 1 of Example 2, compounds 7-1 and 7-2 were separated by the same process as shown in FIG. 1 .

In more detail, the ethyl acetate fraction (14 g) was adsorbed on silica gel, and then a hexane-ethyl acetate-water solution mixed in a volume ratio of 100:0:2 to 0:100:2 was sequentially eluted to obtain 8 fractions ( E1, E2, E3, E4, E5, E6, E7 and E8). After adding the active fraction E4 (1.3 g) to a Biotage SNAP KP-Sil (50 g) column, dichloromethane-acetone solution was eluted under gradient conditions of 0%-50% (v/v), and 10 fractions (E4A, E4B) , E4C, E4D, E4E, E4F, E4G, E4H, E4I and E4J). The active fraction E4I (223.6 mg) was loaded onto a YMC-Pack ODS-A column (20×250 mm, 5 μm) connected to a Shimadzu Prominence HPLC system, and eluted with a 67% aqueous methanol solution at a flow rate of 5 ml/min to obtain compound 7-1 (17.2). mg) and compound 7-2 (27.1 mg) were isolated.

< Example 4> Identification of the chemical structure of bactericidal active compounds

In order to identify the chemical structure of the bactericidal active compound isolated in Example 3, the following experiment was performed.

Specifically, nuclear magnetic resonance (NMR) and mass spectroscopy (MS) were mainly used to elucidate the chemical structure of the compound isolated from the organic solvent fraction of Pterokarya tonkinensis. After dissolving the sample in CDCl ₃ , CD ₃ OD, or C ₅ D ₅ N (Cambridge Isotope Laboratories), ¹ H-NMR, ¹³ C-NMR and 2D-NMR spectra were obtained on a Bruker AMX-400 or AMX-500 instrument. Based on the peak of tetramethylsilane (TMS) added to the NMR solvent, a chemical shift value was expressed as δ ( ppm). The chemical positions of the peaks in the spectrum were determined by combining the NMR data shown in the literature and the experimental data of ¹ H- ¹ H COSY, HSQC, HMBC and DEPT. The mass of the isolated compound was measured using ESI-MS. The chemical formula of the isolated compound was completed using high-resolution mass spectroscopy (Synapt G2 HDMS Q-TOF). In addition, the specific optical rotation of the separated compound was measured with a polarimeter (Auto Digital Polarieter, Rudolph Research Analytical) and an infrared analyzer (FT-IR spectroscopy; Identify IR, Smiths).

<4-1> Identification of the chemical structure of compound 1

As a result of the analysis, in compound 1, a cation of m/z 349 [M + Na] ⁺ in ESI-MS in cation mode was detected as an anion of m/z 325 [M - H] ^- in ESI-MS in anion mode, was determined as C ₂₀ H ₂₂ O ₄ . The NMR analysis results are shown in Table 3 below.

As shown in Table 3 below, 20 carbon signals appeared in the ¹³ C-NMR spectrum, of which 19 constitute diarylheptanoid, and the remainder is an O-methyl group. ¹ H-NMR and ¹³ C-NMR data of compound 1 were compared with the literature (Journal of Natural Products 2002 65:1707-1708), and the isolated compound 1 was prepared as a diarylheptanoid compound 7-(4) It was identified as "-hydroxy-3"-methoxyphenyl)-1-(4'-hydroxyphenyl)-4E-hepten-3-one.

[Formula 1]

Position δ _C ^a , type δ _H , multiple ( J ) One 30.7, CH ₂ 2.79, m 2 42.7, CH ₂ 2.74, m 3 202.8, C - 4 131.6, CH 6.05, dt (15.9, 1.5) 5 149.2, CH 6.87, dt (15.9, 6.9) 6 35.7, CH ₂ 2.48, m 7 35.0, CH ₂ 2.66, t (7.5) One' 133.2, C 2', 6' 130.3, (CH) ₂ 6.96, d (8.5) 3', 5' 116.1, (CH) ₂ 6.67, d (8.4) 4' 156.6, C - One" 133.8, C - 2" 113.1, CH 6.74, d (2.0) 3” 148.9, C - 4" 145.8, C - 5” 116.1, CH 6.69, d (8.0) 6” 121.8, CH 6.60, dd (8.0, 2.0) 3”-OCH ₃ 56.3, CH ₃ 3.79, s

( ^a Recorded in CDCl ₃ at 125 and 500 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-2> Identification of the chemical structure of compound 2-1

As a result of the analysis, compound 2-1 was detected as an anion of m/z 311 [M - H] ^- by ESI-MS in anion mode, and the chemical formula was determined as C ₁₉ H ₂₀ O ₄ . The NMR analysis results are shown in Table 4 below.

As shown in Table 4 below, 19 carbon signals constituting diarylheptanoid were shown in the ¹³ C-NMR spectrum. ¹ H-NMR and ¹³ C-NMR data of compound 2-1 were compared with the literature (Chin. Chem. Lett. 2005, 16:215-218) to prepare the isolated compound 2-1 with diarylhep of the following formula 2-1 The diarylheptanoid compound was identified as pterocarine.

[Formula 2-1]

Position δ _C ^a , type δ _H , multiple ( J ) One 121.5, CH 6.80, dd (8.1, 2.0) 2 123.8, CH 6.87, d (8.2) 3 141.5, C - 4 149.9, C - 5 118.4, CH 6.88, d (1.5) 6 139.8, C - 7 35.2, CH ₂ 2.68, m 8 26.8, CH ₂ 1.69, m 9 18.9, CH ₂ 1.55, m 10 46.0, CH ₂ 1.93, m 11 211.7, C - 12 40.9, CH ₂ 2.33, m 13 26.9, CH ₂ 2.80, m 14 133.0, C - 15 121.3, CH 6.57, dd (8.1, 2.2) 16 115.6, CH 6.74, d (8.1) 17 143.4, C - 18 148.0, C - 19 112.5, CH 5.61, d (2.0)

( ^a Recorded in CDCl ₃ at 100 and 400 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-3> Identification of the chemical structure of compound 2-2

As a result of the analysis, compound 2-2 was detected as a cation of m/z 349 [M + Na] ⁺ in ESI-MS in cation mode, and the chemical formula was determined as C ₂₀ H ₂₄ O ₅ . The NMR analysis results are shown in Table 5 below.

As shown in Table 5 below, 19 carbon signals and one O-methyl carbon signal constituting diarylheptanoid were shown in the ¹³ C-NMR spectrum. By comparing the hydrogen and ¹³ C-NMR data of compound 2-2 with the literature (Tetrahedron Letters 1976, 17:3069-3072), the isolated compound 2-2 was converted into a diarylheptanoid compound galeon of the following formula 2-2 sympathized.

[Formula 2-2]

Position δ _C ^a , type δ _H , multiple ( J ) One 27.5, CH ₂ 2.96, dd (16.2, 11.1); 2.70, dd (16.2, 8.4) 2 41.5, CH ₂ 2.35, dd (16.6, 10.2); 2.24, dd (16.5, 8.5) 3 210.4, C - 4 46.5, CH ₂ 1.95, m; 1.55, m 5 19.2, CH ₂ 1.63, m; 1.55, m 6 27.5, CH ₂ 1.77, dd (13.6, 6.9); 1.55, m 7 36.1, CH ₂ 2.82, dd (12.2, 5.6); 2.63, m One' 133.4, C - 2' 112.4, CH 5.55, s 3' 147.4, C - 4' 143.3, C - 5' 115.1, CH 6.82, d (8.1) 6' 122.1, CH 6.60, d (8.0) One" 140.2, C - 2" 115.2, CH 6.86, overlapped 3” 152.3, C 4" 143.0, C - 5” 124.2, CH 7.00, d (8.5) 6” 122.2, CH 6.86, overlapped 3”-OCH ₃ 56.2, CH ₃ 3.71, s

( ^a Recorded in CDCl ₃ at 125 and 500 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-4> Identification of the chemical structure of compound 2-3

As a result of the analysis, compound 2-3 was detected as a cation of m/z 365 [M + Na] ⁺ in ESI-MS in cation mode, and the chemical formula was determined as C ₂₀ H ₂₄ O ₆ . The NMR analysis results are shown in Table 6 below.

As shown in Table 6 below, 19 carbon signals and one O-methyl carbon signal constituting diarylheptanoid were shown in the ¹³ C-NMR spectrum. Hydrogen and ¹³ C-NMR data and nonlinear measurement data of compound 2-3 [α] _D ^25.4 = -123.0 (c = 0.1, CHCI ₃ ) were obtained from the literature (Chem. Pharm. Bull. 2003, 51:262-264) and The comparatively isolated compound 2-3 is a diarylheptanoid compound of Formula 2-3 below 3',4"-epoxy-1-(4'-hydroxylphenyl)-7-(3"-methoxyl-phenyl)- It was identified as heptane-2-hydro-3-one.

[Formula 2-3]

Position δ _C ^a , type δ _H , multiple ( J ) One 37.9, CH ₂ 2.91, d (4.2) 2 77.4, CH 4.08, d (4.6) 3 211.6, C - 4 40.0, CH ₂ 1.94, ddd (17.4, 11.8, 5.6); 1.16, m 5 20.8, CH ₂ 1.70, m; 1.50, m 6 28.5, CH ₂ 1.86, m; 1.43, m 7 36.6, CH ₂ 2.86, m;2.57, m One' 126.5, C - 2' 117.5, CH 5.82, d (2.1) 3' 147.8, C - 4' 145.6, C - 5' 115.4, CH 6.83, d (8.1) 6' 125.2, CH 6.71, dd (8.1, 2.1) One" 140.4, C - 2" 115.3, CH 6.83, overlapped 3” 151.6, C - 4" 145.9, C - 5” 124.9, CH 7.19, d (8.5) 6” 120.6, CH 6.83, overlapped 3”-OCH ₃ 55.9, CH ₃ 3.79, s

( ^a Recorded in CDCl ₃ at 125 and 500 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-5> Identification of the chemical structure of compound 3

As a result of the analysis, compound 3 was detected as a cation of m/z 255.1000 [M + Na] ⁺ in HR-ESI-MS in cation mode, and the chemical formula was determined as C ₁₄ H ₁₆ O ₃ (Calc. m/z 255.0997). . The NMR analysis results are shown in Table 7 below.

As shown in Table 7 below, 14 carbon signals constituting norsesquiterpene lactone were shown in the ¹³ C-NMR spectrum. In the hydrogen NMR spectrum, δ _H 7.48 (d, H-8), 6.87 (d, H-7) ppm of two sp ² methyl hydrogens, δ _H 5.10 (d, H-1), 2.87 (dd, H-4), 2.63 ( 6 sp ³ methyl hydrogens at dd, H-4), 2.09 (m, H-3), 2.03 (td, H-11), 1.71 (m, H-3), 1.47 (td, H-2) ppm , δ _H 1.10 (d, H-12) and 1.07 (d, H-13) ppm of two sp ³ methyl hydrogen signals were shown. ¹³ In the C-NMR spectrum, δ _C 1 carboxyl carbon at 171.8 ppm, δ _C 160.3, 152.7, 119.5, 114.5 ppm quaternary sp ² 4 carbons, δ _C 124.1, 116.2 ppm of sp ² methyl carbon 2 carbons, δ _C 3 sp ³ methyl carbons at 80.4, 44.7, 29.3 ppm, δ _C 22.4, 21.5 ppm sp ³ methylene carbon 2, δ _C Two sp ³ methyl carbons of 18.7 and 17.9 ppm were found.

¹ H-NMR, ¹³ C-NMR, ¹ H- ¹³ C HMBC data and nonlinear measurement data of compound 3 [α] _D ^24.0 = -92.0 (c = 0.1, CH ₃ OH) were described in the literature (Phytochemistry 1986, 25:1923). -1926) and the norsesquiterpene lactone compound (8R-cis)-6,7,8,8a-tetrahydro-5-hydroxy-8-(1- It was identified as methylethyl)-2H-naphtho[1,8-bc]furan-2-one.

[Formula 3]

Position δ _C ^a , type δ _H , multiple ( J ) One 80.4, CH 5.10, d (11.1) 2 44.7, CH 1.47, td (12.1, 3.5) 3 22.4, CH ₂ 2.09, m; 1.71, m 4 21.5, CH ₂ 2.87, dd (18.1, 7.6); 2.63, dt (18.0, 8.8) 5 119.5, C - 6 160.3, C - 7 116.2, CH 6.87, d (8.2) 8 124.1, CH 7.48, d (8.2) 9 114.5, C - 10 152.7, C - 11 29.3, CH 2.03, td (7.0, 4.0) 12 18.7, CH ₃ 1.10, d (6.9) 13 17.9, CH ₃ 1.07, d (6.9) 14 171.8, C

( ^a Recorded in CD ₃ OD at 100 and 400 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-6> Identification of the chemical structure of compound 4

The results of NMR analysis of Compound 4 are shown in Table 8 below.

As shown in Table 8 below, 13 carbon signals constituting dinorsesquiterpene were shown in the ¹³ C-NMR spectrum. By comparing the NMR data of compound 4 with the literature (J. Agric. Food Chem. 1990, 38:2045-2049), the dinorsesquiterpene compound of the following formula 4 was 9-hydroxymegastigm-7-en-3- identified as one.

[Formula 4]

Position δ _C ^a , type δ _H , multiple ( J ) One 38.3, C - 2 56.0, CH ₂ 2.31, d (13.4); 2.14, dd (13.3, 2.4) 3 210.9, C - 4 49.4, CH ₂ 2.07, t (5.8); 2.04, t (6.4) 5 33.4, CH 1.91, m 6 56.9, CH 1.88, m 7 129.1, CH 5.39, dd (15.4, 8.4) 8 138.3, CH 5.64, dd (15.4, 6.4) 9 68.8, CH 4.37, pd (6.4, 1.1) 10 23.7, CH ₃ 1.32 d (6.4) 5-CH ₃ 21.3, CH ₃ 0.94, d (5.9) 1 _ax -CH ₃ 21.1, CH ₃ 0.84, s 1 _eq -CH ₃ 30.6, CH ₃ 1.00, s

( ^a Recorded in CDCl ₃ at 100 and 400 MHz for ¹³ C and ¹ H NMR and; δ in ppm and J in Hz)

<4-7> Identification of the chemical structure of compound 5-1

The results of NMR analysis of compound 5-1 are summarized below.

compound 5-1; ¹ H-NMR (500 MHz, CDCl ₃ ) 8.59(OH, br s), 7.58(1H, d, 7.7), 7.29(1H, td, 7.9, 1.6), 7.09(1H, d, 8.0), 4.99( 1H, dd, 9.3, 4.1), 3.57 (3H, d, 1.7), 2.87 (1H, m), 2.53 (2H, m), 2.22 (1H, m).

By comparing the hydrogen NMR data of compound 5-1 with the literature (Chem. Pharm. Bull. 2007 55:1316-1318), the separated compound 5-1 is bicyclic aromatic hydrocarbon of the following formula 5-1 ) was identified as the compound xylarenone, and it was confirmed that it was a racemic mixture based on nonlinear measurement data [α] _D ^26.7 = 0 (c = 0.1, CHCl ₃ ).

[Formula 5-1]

<4-8> Identification of the chemical structure of compound 5-2

The results of NMR analysis of compound 5-2 are summarized below.

compound 5-2; ¹ H-NMR (500 MHz, CDCl ₃ ) 12.10 (OH, br s), 7.08 (1H, d, 9.0), 6.86 (1H, d, 9.0), 4.93 (1H, dd, 9.9, 4.2), 3.57 ( 3H, s), 2.88 (1H, dt, 17.2, 4.8), 2.60 (1H, m), 2.47 (1H, dt, 13.4, 5.2), 2.20 (1H, m).

By comparing the hydrogen NMR data of compound 5-2 with the literature (Phytochemistry 1996, 42:173-176), the separated compound 5-2 is a bicyclic aromatic hydrocarbon compound 5 of the following formula 5-2, It was identified as 8-dihydroxy-4-methoxy-α-tetralone, and it was confirmed that it was a racemic mixture based on nonlinear measurement data [α] _D ^27.3 = 0 (c = 0.1, CHCl ₃ ).

[Formula 5-2]

<4-9> Identification of the chemical structure of compound 6

The results of NMR analysis of compound 6 are summarized below.

compound 6; ¹ H-NMR (500 MHz, CDCl ₃ ) 7.64(1H, d, 15.8), 7.42(2H, d, 8.2), 6.85(2H, d, 8.2), 6.30(1H, d, 15.9), 3.80(3H) , s); ¹³ C-NMR (125 MHz, CDCl ₃ ) 168.0, 157.9, 144.7, 130.0, 127.1, 115.9, 115.1, 51.7.

By comparing the hydrogen NMR data of compound 6 with the literature (Journal of Natural Products 1988, 51:588-590), the separated compound 6 is an aromatic hydrocarbon compound of the following formula 6 ( E )-4-hydroxycinnamic acid methyl Identified as ester.

[Formula 6]

<4-10> Identification of the chemical structure of compound 7-1

The results of NMR analysis of compound 7-1 are summarized below.

compound 7-1; ¹ H-NMR (C ₅ D ₅ N, 500 MHz): δ _H 5.48 (1H, t, J = 3.6 Hz; H-12), 4.22(3H, overlapped; H-2, H-3, and H- 23), 3.74 (1H, d, J = 10.5 Hz; H-23), 1.16 (3H, s; H-27), 1.08 (6H, s; H-24, H-25, and H-26), 0.98 (3H, d, J = 6.4 Hz; H-29), 0.94 (3H, d, J = 6.4 Hz; H-30); ¹³ C-NMR (C ₅ D ₅ N, 125 MHz), 179.7 (C-28), 139.1 (C-13), 125.3 (C-12), 78.0 (C-3), 68.6 (C-2), 66.3(C-23), 53.3(C-18), 47.9(C-5), 47.8(C-17), 47.7(overlapped; C-1 and C-9), 43.4(C-4), 42.2( C-14), 39.8 (C-8), 39.2 (overlapped; C-19 and C-20), 38.1 (C-10), 37.2 (C-22), 33.0 (C-7), 30.8 (C- 21), 28.4 (C-15), 24.7 (C-16), 23.7 (C-27), 23.6 (C-11), 21.1 (C-30), 18.3 (C-6), 17.3 (overlapped; C) -25, C-26, and C-29), 14.2 (C-24).

¹ H-NMR and ¹³ C-NMR data of compound 7-1 were compared with the literature (Molecules 2008, 13:2717-2728), and the isolated compound 7-1 was prepared with ursane triterpene of the following formula 7-1 triterpene) was identified as the compound asiatic acid.

[Formula 7-1]

<4-11> Identification of chemical structure of compound 7-2

The results of NMR analysis of compound 7-2 are summarized below.

compound 7-2; ¹ H-NMR (C ₅ D ₅ N, 500 MHz): δ _H 5.45 (1H, t, J = 3.5 Hz; H-12), 4.21(2H, overlapped; H-2 and H-3), 4.18( 1H, overlapped; H-23), 3.70(1H, d, J = 10.4 Hz; H-23), 1.18(3H, s; H-27), 1.06(3H, s; H-25), 1.05(3H) , s; H-24), 1.02 (3H, s; H-26), 0.97 (3H, s; H-29), 0.90 (3H, s; H-30); ¹³ C-NMR (C ₅ D ₅ N, 125 MHz) 180.1 (C-28), 144.7 (C-13), 122.2 (C-12), 78.0 (C-3), 68.6 (C-2), 66.3 (C-23), 48.0 (C-9), 47.7 (C-5), 47.5 (C-1), 46.4 (C-17), 46.2 (C-19), 43.4 (C-4), 42.0 ( C-14), 41.8 (C-18), 39.6 (C-8), 38.2 (C-10), 34.1 (C-21), 33.0 (overlapped; C-22 and C-29), 32.7 (C- 7), 30.7 (C-20), 28.1 (C-15), 25.9 (C-27), 23.7 (C-16), 23.5 (overlapped; C-11 and C-30), 18.3 (C-6) , 17.3 (C-26), 17.1 (C-25), 14.1 (C-24).

¹ H-NMR and ¹³ C-NMR data of compound 7-2 were compared with the literature (Molecules 2008, 13:2717-2728), and the isolated compound 7-2 was prepared with oleanane of the following formula 7-2. triterpene) was identified as the compound arjunolic acid.

[Formula 7-2]

<4-12> Identification of the chemical structure of compound 7-3

The results of NMR analysis of compound 7-3 are summarized below.

compound 7-3; 1H-NMR (CD ₃ OD, 400 MHz) δ _H 5.27 (1H, t, J = 3.5 Hz, H-13), 3.62 (1H, m, H-2), 2.91 (1H, d, J = 10.0 Hz) ; H-3), 1.17 (3H, s; H-23), 1.02 (3H, s; H-30), 1.01 (3H, s; H-24), 0.95 (3H, s; H-30), 0.91 (3H, s; H-25), 0.81 (3H, s; H-29).

By comparing the hydrogen NMR data of compound 7-3 with the literature (Chem. Pharm. Bull. 1978 26:3075-3079), the isolated compound 7-3 is an oleanane triterpene compound of the following formula 7-3 It was identified with maslinic acid.

[Formula 7-3]

<4-13> Identification of the chemical structure of compound 8

The results of NMR analysis of compound 8 are summarized below.

compound 8; ¹ H-NMR (CDCl ₃ , 400 MHz): δ _H 4.67 (1H, d, J = 2.4 Hz, H-29), 4.55 (1H, dd, J = 2.6, 1.5 Hz, H-29), 3.42 (1H) , dd, J = 11.3, 4.6 Hz, H-7), 3.24 (1H, dd, J = 12.1, 4.4 Hz, H-3), 1.67 (3H, s, H-30), 1.04 (3H, s, H-25), 0.95 (3H, s, H-28), 0.94 (3H, s, H-23), 0.90 (3H, s, H-26), 0.79 (3H, s, H-27), 0.75 (3H, s, H-24).

The hydrogen NMR data of compound 8 was compared with the literature (Phytochemistry 1973, 12:3004-3006), and the isolated compound 8 was identified as a lupane triterpene compound loranthol of Formula 8 below.

[Formula 8]

< Example 5> containing the isolated compound plant disease Evaluation of antibacterial activity against phytopathogenic fungi of the control composition

In order to investigate the bactericidal activity spectrum of the compounds 1 to 8 isolated in Example 3 against plant pathogens, the following experiment was performed, and the results are shown in Table 9.

Specifically, the plant pathogen Alternaria brassicola (Alternaria brassicicola), Coletotricum cocodes (Colletotrichum coccodes), Fusarium oxysporum (Fusarium oxysporum), Magnaporte Orize (Magnaporthe oryzae) and Phytophthora Infestans (Phytophthora infestans) was evaluated by the broth micro-dilution method using a 96-well plate to determine the minimum inhibitory concentration (MIC) value. Alternaria brassicola (Alternaria brassicicola) and Fusarium Oxyforum (Fusarium oxysporum) was inoculated on potato agar medium, inoculated on potato agar medium, and spores formed by culturing at 25°C for 10 days were used for the experiment. Phytophthora Infestance (Phytophthora infestans) was inoculated on oatmeal agar medium and then cultured for 10 days at 20°C. The spores formed were used for the experiment. Coletotricum cocodes (Colletotrichum coccodes) and Magnaporte Orize (Magnaporthe oryzae) was inoculated on an oatmeal agar medium and cultured at 25° C. for 7 days, and then mycelia were removed. Then, spore formation was induced by light treatment for 2 days in a constant temperature and humidity room at 25°C and 80% relative humidity. For all strains used in the experiment, spores were harvested using a potato liquid medium, and a spore suspension was prepared by filtering it with a 4-fold gauze.

Finally, the spores of plant pathogens were included in 100 μl wells at a concentration of 1×10 ⁴ spores/ml, and compounds 1 to 8 were 1, 2, 4, 8, 16, 32, 63, 125, 250, 500 μg/ It was dispensed to be included at a concentration of ml, and the content of methanol at this time did not exceed 1%. The addition of 1% methanol was used as a control, and the experiment was repeated 3 times for each concentration. After culturing them at 20 °C or 25 °C for 2 to 3 days, the concentration at which the growth of mold was completely inhibited was determined as the minimum inhibitory concentration (MIC).

plant pathogens MIC (μg/ml) compound 1 compound 2-1 compound 2-2 compound 2-3 compound 3 compound 4 compound 5-1 compound 5-2 compound 6 compound 7-1 compound 7-2 compound 7-3 compound 8 Alternaria brassicicola 125 - - - - - - - 500 125 - - - Colletotrichum coccodes 125 - 500 - - - - 500 250 500 - - - Fusarium oxysporum 500 - - - 500 - - - 500 500 - - - Magnaporthe oryzae - - 250 - 500 250 - 125 250 63 16 500 500 Phytophthora infestans 31 125 500 500 - - 500 500 250 - - - -

As shown in Table 9 above, Compound 1 was Phytophthora at a concentration of 31 μg/ml ( Phytophthora ). infestans ; Potato / tomato blight) showed excellent activity to completely inhibit the growth, and at a concentration of 125 μg/ml, Alternaria brassicicola ( Alternaria brassicicola ; Brassica black blight) and Coletotrichum coccodes ; The growth of pepper anthrax) was completely inhibited, and Fusarium oxysporum ( Fusarium ) at a concentration of 500 μg/ml oxysporum ; It showed bactericidal activity to completely inhibit the growth of wilt bacteria).

Compound 2-1 is Phytophthora infestans ( Phytophthora at a concentration of 125 μg / ml) infestans ; Potato / tomato late blight) showed excellent activity to completely inhibit the growth, and Compound 2-2 at a concentration of 250-500 μg / ml Colletotrichum coccodes (Capsicum anthrax), Magnaforte olease ( Magnaporthe oryzae ; Rice blast bacillus) and Phytophthora infestans ( Phytophthora infestans ; potato / tomato late blight) were completely inhibited, and Compound 2-3 was Phytophthora infestans ( Phytophthora ) at a concentration of 500 μg/ml. infestans ; Potato/tomato late blight) showed bactericidal activity to completely inhibit the growth.

In addition, compound 3 is Fusarium oxysporum at a concentration of 500 μg / ml oxysporum ; It showed bactericidal activity against wilting bacteria) and Magnaporthe oryzae (rice blast bacteria), and compound 4 completely inhibited the growth of Magnaporthe oryzae (rice blast bacteria) at a concentration of 250 μg/ml, Compound 5-1 exhibited bactericidal activity to completely inhibit the growth of Phytophthora infestans (potato/tomato late blight) at a concentration of 500 μg/ml.

In addition, compound 5-2 at a concentration of 125 μg /ml oryzae ; It exhibited a strong bactericidal activity to completely inhibit the growth of rice blast bacillus), and at a concentration of 500 μg/ml, Colletotrichum coccodes ; chilli anthrax) and Phytophthora infestans ; Potato/tomato late blight) was completely inhibited.

Compound 6 can be used at a concentration of 250-500 μg/ml in Alternaria brassicola. brassicicola ; Brassica crops black-spotted fungus), Colletotrichum coccodes ; Chili anthrax), Fusarium oxysporum oxysporum ; wilting bacteria), Magnaporthe oryzae ; Rice Blast Bacteria) and Phytophthora infestans ; Potato/tomato late blight) was completely inhibited.

Compound 7-1 is Magnaporthe olase ( Magnaporthe ) at a concentration of 63-125 μg/ml oryzae ; It exhibited strong bactericidal activity to completely inhibit the growth of rice blast bacillus) and Alternaria brassicicola (Brassicaceae black spot blight), and at a concentration of 500 μg/ml, Colletotrichum coccodes ; chilli anthrax) and Fusarium oxysporum oxysporum ; It showed bactericidal activity to completely inhibit the growth of wilt bacteria).

Compound 7-2 at a concentration of 16 μg / ml Magnaporthe olase ( Magnaporthe oryzae ; It showed strong bactericidal activity against rice blast bacteria), and compounds 7-3 and 8 were Magnaporthe at a concentration of 500 μg/ml. oryzae ; It showed bactericidal activity to completely inhibit the growth of rice blast bacteria).

< Example 6> containing the isolated compound plant disease Control composition against phytopathogenic bacteria antibacterial activity evaluation

In order to investigate the bactericidal activity spectrum of the compounds 1 to 8 isolated in Example 3 against phytopathogenic bacteria, the following experiment was performed, and the results are shown in Table 10.

Specifically, the activity of inhibiting the growth of plant bacterial pathogenic bacteria was evaluated using 13 types of compounds isolated from the Pterokarya tonkinensis extract. Each of the 13 separated compounds was dissolved in methanol at a concentration of 100 mg/ml, and then diluted in methanol by a double dilution method to prepare stock solutions for each concentration. Using a 96-well plate, each isolated compound was finally added to 99 μl of tryptic soy broth (TSB) medium at 1, 2, 4, 8, 16, 32, 63, 125, 250, 500 μg/ml per well. 1 μl of the stock solution for each concentration was dispensed to obtain a concentration, and the content of methanol did not exceed 1% at this time. 10 phytobacterial pathogenic bacterium Exidoborax avene subsp. Cattle ( Acidovorax avenae ) subsp. cattleyae ; Phalaenopsis orchid bacterial brown spot disease), Agrobacterium tumefaciens ( Agrobacterium tumefaciens ; fruit tree root-knot), Clavibacter misiganensis subsp. Mishiganensis ( Clavibacter michiganensis subsp. michiganensis ; The antibacterial activity against red pepper ulcer disease) was evaluated by obtaining the minimum inhibitory concentration (MIC) value, which is the minimum inhibitory concentration, by the broth micro-dilution method using a 96-well plate. After inoculation in TSB medium, the culture was shaken at 30° C. for 2 days at 150 rpm, and then diluted with TSB medium to obtain a bacterial concentration of 1×10 ⁷ CFU/ml. The prepared bacterial culture solution was inoculated, and finally 1×10 ⁵ CFU/ml was made. A non-treated group with only 1% methanol was used, and the concentration at which bacterial growth was completely inhibited (MIC, minimum inhibitory concentration) was determined after culturing for 2 to 3 days.

plant pathogens MIC (μg/ml) compound 1 compound 2-1 compound 2-2 compound 2-3 compound 3 compound 4 compound 5-1 compound 5-2 compound 6 compound 7-1 compound 7-2 compound 7-3 compound 8 Acidovorax avenae subsp. cattleyae 63 125 500 500 - - - 500 500 8 16 - - Agrobacterium tumefaciens 500 500 - - - - - - 500 - - - - Clavibacter michiganensis subsp. michiganensis 500 500 - - - - - - 500 - - - -

As shown in Table 10, Compound 1 and Compound 2-1 were obtained from Exidovorax avene subsp. at a concentration of 63-125 μg/ml. Cattle ( Acidovorax avenae ) subsp. cattleyae ; It exhibited a strong activity to completely inhibit the growth of Phalaenopsis orchid bacteria, and Agrobacterium tumefaciens ( Agrobacterium tumefaciens) at a concentration of 500 μg/ml tumefaciens ; fruit tree root-knot) and Clavibacter misiganensis subsp. Mishiganensis ( Clavibacter michiganensis subsp. michiganensis ; It exhibited bactericidal activity that completely inhibited the growth of red pepper ulcer disease).

Compound 7-1 and compound 7-2 were obtained from Exidovorax avene subsp. at a concentration of 8-16 μg/ml. Cattle showed a strong activity to completely inhibit the growth of ( Acidovorax avenae subsp. cattleyae ; Phalaenopsis bacterial brown spot disease), and Compound 2-2, Compound 2-3 and Compound 5-2 were exci at a concentration of 500 μg/ml. Boolax Abene subsp. Cattray ( Acidovorax) avenae subsp. cattleyae ; Phalaenopsis orchid showed bactericidal activity to completely inhibit the growth of the bacterial brown spot bacterium).

Compound 6 at a concentration of 500 μg / ml Exidovorax avene subsp. Cattray ( Acidovorax) avenae subsp. cattleyae ; Phalaenopsis orchid bacterial brown spot bacterium), Agrobacterium tumefaciens ( Agrobacterium tumefaciens ; fruit tree root-knot) and Clavibacter misiganensis subsp. Mishiganensis ( Clavibacter michiganensis subsp. michiganensis ; It exhibited bactericidal activity that completely inhibited the growth of red pepper ulcer disease).

< Example 7> containing the isolated compound plant disease of the control composition plant disease Control effectiveness evaluation

In vivo plant diseases of compounds 1, 2-1 to 2-3, and 3, compounds 5-1 and 5-2, and compounds 7-1 to 7-3 among the 13 isolated compounds obtained in Example 3 In order to investigate the control activity, the following experiment was performed. The results are shown in Table 11 below.

Specifically, rice blast, tomato late blight, wheat red of compounds 1, 2-1 to 2-3, and 3, compound 5-2 and compounds 7-1 to 7-3 isolated from the Pterocarya tonkinensis extract. The control effect on rust and pepper anthracnose was investigated under greenhouse conditions. After dissolving the active ingredient in methanol, a surfactant Tween 20 solution (250 μg/ml) was added to adjust the final concentration of the drug to 100 μg/ml, 500 μg/ml or 1000 μg/ml, and the final methanol concentration of all samples was 5% adjusted to At this time, as a control, a solution containing 5% methanol and 250 μg/ml of Tween 20 was used. Four pots were used for each plant bottle, and after spraying the active ingredient sample on the leaf surface, it was air-dried for 24 hours, and then each plant pathogen was inoculated. Control activity against these four plant diseases was investigated according to the method described in Example 1.

sample Concentration (μg/ml) Control Value (%) RCB TGM TLB PAN compound 1 500 25 95 67 35 100 25 0 20 0 compound 2-1 1000 25 100 33 20 500 0 100 0 8 compound 2-2 1000 88 15 0 0 500 25 0 0 0 compound 2-3 500 0 76 0 0 compound 3 500 88 52 7 0 100 63 27 0 0 compound 5-2 500 94 15 0 0 100 56 0 0 0 compound 7-1 500 88 91 0 0 100 38 33 0 0 compound 7-2 500 88 92 0 0 100 38 91 0 0 compound 7-3 1000 63 45 0 0 500 25 52 0 0

(RCB: rice blast disease, TGM: tomato gray mold disease, TLB: tomato late blight, PAN: red pepper anthrax)

As shown in Table 11, when plants were treated with diarylheptanoid compounds 1, 2-1 to 2-3, various effects were shown in controlling rice blast disease, tomato gray mold disease, and tomato late blight. When Compound 1 was treated with plants at a concentration of 500 μg/ml, it showed an effect of controlling 95% and 67% of tomato gray mold and tomato late blight, respectively, and Compound 2-1 was treated with plants at a concentration of 500 μg/ml or more When the plant was treated with compound 2-2 at a concentration of 1000 μg/ml, it exhibited the effect of inhibiting the occurrence of rice blast by 88%. When the compound 2-3 was treated with plants at a concentration of 500 μg/ml, it showed an effect of inhibiting tomato gray mold disease by 76%.

In addition, when plants were treated with norsesquiterpene lactone compound 3 at a concentration of 500 μg/ml, rice blast and tomato gray mold disease were respectively 88% and 52% effective in controlling, bicyclic When plants were treated with aromatic hydrocarbon compound 5-2 at a concentration of 500 μg/ml, the effect of inhibiting the occurrence of rice blast disease by 94% was shown.

When plants were treated with ursane triterpene compound 7-1 and olenane triterpene compound 7-2 at a concentration of 500 μg/ml, the occurrence of rice blast and tomato gray mold disease was reduced. 88% to 92% inhibitory effect was shown. When plants were treated with another olenane triterpene compound 7-3 at a concentration of 1000 μg/ml, the occurrence of rice blast disease and tomato gray mold disease was inhibited by 63% and 45%, respectively. It was similar to the control spectrum shown when the Pterocarya tonkinensis extract was treated, and showed a higher level of control activity.

Although the above has been described with reference to the preferred embodiments of the present invention, those skilled in the art can variously modify and change the present invention without departing from the spirit and scope of the present invention as set forth in the claims below. You will understand that you can.

Claims (16)

A composition for controlling plant diseases comprising at least one selected from the compounds represented by the following formulas 1 to 8:
[Formula 1]

;
[Formula 2]

In the above formula,
R ₁ to R ₃ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl;
[Formula 3]

;
[Formula 4]

;
[Formula 5]

R ₄ to R ₆ are each independently a hydrogen atom, —OH, —OC ₁ -C ₆ alkyl, or C ₁ -C ₆ alkyl;
[Formula 6]

;
[Formula 7]

In the above formula,
R ₇ and R ₈ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —OC ₁ -C ₆ alkyl,
R ₉ and R ₁₀ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, OH or —C ₁ -C ₆ alkyl-OH; and
[Formula 8]

. The composition for controlling plant diseases according to claim 1, comprising at least one selected from compounds represented by the following formulas 1 to 8:
[Formula 1]

;
[Formula 2-1]

;
[Formula 2-2]

;
[Formula 2-3]

;
[Formula 3]

;
[Formula 4]

;
[Formula 5-1]

;
[Formula 5-2]

;
[Formula 6]

;
[Formula 7-1]

;
[Formula 7-2]

;
[Formula 7-3]

; and
[Formula 8]

. According to claim 1,
The compound is a composition for controlling a plant disease having a bactericidal activity. According to claim 1,
The plant disease is a plant disease control composition that is caused by a phytopathogenic fungus or phytopathogenic bacteria. 5. The method of claim 4,
The phytopathogenic fungus is Alternaria brassicola . brassicicola ; Brassica crops black spot fungus), Botrytis cineria ( Botrytis ) cinerea ; gray mold fungus), Colletotrichum coccodes ; Chili anthrax), Fusarium oxysporum (wilt), Magnaporthe oryzae ; Rice Blast Bacteria), Puccinia triticina (Wheat Red Rust Bacteria) and Phytophthora Phytophthora infestans ; Potato / tomato late blight) is selected from the group consisting of a plant disease control composition. 5. The method of claim 4,
The phytopathogenic bacteria are Exidoborax avene subsp. Cattray ( Acidovorax) avenae subsp. cattleyae ; Phalaenopsis orchid bacterial brown spot bacterium), Agrobacterium tumefaciens ( Agrobacterium tumefaciens ; fruit tree root-knot), Clavibacter misiganensis subsp. Clavibacter michiganensis subsp. michiganensis ; A composition for controlling plant diseases that is selected from the group consisting of red pepper ulcer disease). According to claim 1,
The compounds represented by Formulas 1 to 8 are Pterocarya tonkinensis ( Pterocarya 　 tonkinensis ), which is derived from, a composition for controlling plant diseases. 8. The method of claim 7,
The compounds represented by Formulas 1 to 8 are Pterocarya tonkinensis ( Pterocarya 　 tonkinensis ) of an extract or a fraction thereof, a composition for controlling plant diseases. 9. The method of claim 8,
The extract is selected from the group consisting of water, C ₁ to C ₄ lower alcohol, hexane, chloroform, methylene chloride, ethyl acetate, acetone, acetonitrile, and combinations thereof. Composition for controlling plant diseases. 9. The method of claim 8,
The fraction of the extract is any one selected from the group consisting of hexane, chloroform, ethyl acetate, methylene chloride, methanol, ethanol, butanol, water, and combinations thereof or a mixture thereof as a solvent. Plant disease fractionated using Control composition. The composition for controlling plant diseases according to claim 1, wherein the compound is included in the composition for controlling plant diseases at a concentration of 100 to 5000 μg/ml. A plant disease control method comprising; treating a plant, its seed, or its habitat with the composition for controlling plant disease of claim 1 . The method according to claim 12, wherein the control method is carried out by a volatilization treatment method or a seed treatment method. The method according to claim 12, wherein the control method is to treat the composition for controlling plant diseases on the surface of the stems and leaves of plants, sterilizing seeds and coating, or in the habitat in which the plants grow. 13. The method of claim 12, wherein the habitat is a planting hole, a furrow, a perimeter of a replantation, a planting furrow, the entire surface of a growth site, a part between the soil and a plant, a part between the roots, a part under the stem of a plant. A control method that is selected from the group consisting of site, main furrow, growing soil, nail bed, box for seedling, tray for seedling, or seedbed. The control method according to claim 12, wherein the treatment is performed during the simulated cultivation period of the composition, before cultivation settlement, at the time of cultivation settlement, and during the growth period after cultivation settlement.

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