KR20190009901A - Composition for controlling plant diseases comprising pharbitin isolated from Pharbitis nil as effective component and method for controlling plant diseases using the same - Google Patents

Abstract

The present invention relates to a composition for controlling plant diseases, containing pharbitin isolated from pharbitidis semen as an active ingredient and a method for controlling plant diseases using the same. According to the present invention of the pharbitin, the composition is harmless to the human body and exhibits excellent control activity against a Xanthomonas bacterium and a plant disease caused by a Ralstonia solanacearum bacterium without causing environmental pollution, thereby being useful in the development of biological pesticides, organic agricultural materials, and production of environmentally friendly grains.

Description

[0001] The present invention relates to a composition for controlling plant diseases, which contains parvitin isolated from black shrubs as an active ingredient, and a method for controlling plant diseases using the same.

The present invention relates to a plant disease control composition containing parvitin isolated from black shafts as an active ingredient, and a method for controlling plant diseases using the same.

Among plant pathogens, germs are a major cause of plant diseases and cause plant diseases in crops worldwide, resulting in reduced yields. In addition, warming due to climate change has been accompanied by an ongoing increase in the incidence of bacterial botanical diseases, and thus ten major phytopathogenic bacteria have been reported based on economics and science (Mansfield et al . 2012, Mol. Plant Pathol 13: 614-629). (1) Pseudomonas ache dogs lost bars (Pseudomonas syringae pathovars), (2) Central Stony Oh Solana Seah room (Ralstonia solanacearum), (3) Agrobacterium tyumeo Pacific Enschede (Agrobacterium tumefaciens , (4) Xanthomonas oryzae pv. oryzae , (5) Xanthomonas campestris pathovars, (6) Zanthomonas axonopodisops bars Xanthomonas axonopodis pathovars), (7) Erwinia amilova amylovora ), (8) Xylella fastidiosa , (9) Dickeya , and (10) Pectobacterium carotovora ). Among the 10 major phytopathogenic bacteria, the three species belonging to the genus Zanthomonas are the major phytopathogenic bacteria. Especially the Zanthomonas Arbor Ricola PV. Rooney program (Xanthomonas arboricola pv. pruni, (Xap)) is a major invasive plant pathogenic bacteria has a wide host of stone fruit including peach, nectarine, plum, plums and the like. In 2010, damage caused by Xap in plum orchards in northern Italy was 11,200 euros per hectare, and the loss of plum, peach and nectar yields in Japan was up to 10,000 euros per hectare. Domestically, peach is the top 5 income source of farm households, and production and cultivation area are increasing every year, so Xap is highly demanded.

Ralstonia solanacearum ) is a soil and waterborne pathogenic bacterium that causes bacterial foot blight in over 200 plants, including tomatoes, potatoes, peppers, peanuts, tobacco, branches and bananas, greatly affecting global crop production. It is also difficult to control bacterial foot rot, and thus the use of crops resistant to pathogens is the most effective method. So far, many commercial antibiotics, copper compounds and resistant insecticides have been used to control phytopathogenic bacteria, but repeated use for long periods has caused environmental pollution, residual toxicity, appearance of resistant bacteria. Therefore, it is necessary to develop natural bio-pesticides that use plant and microbial-derived materials that can replace synthetic bactericides as a new leading material.

On the other hand, the black shark (Pharbitidis Semen) is also called as a mudflapper, and the medicinal medicine name is Pharbitidis Semen and its scientific name is Pharbitis nil or Ipomoea nil , and it has been used in civilization and in the past since it has been used in sub-Saharan Africa, large insecticide, aqua exorcism, diarrhea, wet heat accumulation, secretion of stool, alluvial abdomen, swelling, abdominal distension and constipation. It is a seed of Pharbitis nil Choisy which grows up to about 3m from the stem to the left and has a downward hairs on the stem. In July ~ August, the flowers bloom in various colors such as reddish purple, white, and red. One to three flowers hang on the axilla. The corolla is 10 ~ 23mm in diameter with funnel shape, and the bud has the wrinkle which is like the tip of the brush and dries to the right. There are also five stamens and one pistil. The fruit has two seeds in each of three chambers and ripens in September and October.

Korean Patent Laid-Open No. 2009-0110401 discloses a pharmaceutical composition for preventing or treating periodontal disease, which comprises an insect extract. Korean Patent No. 1478410 discloses a pharmaceutical composition for preventing or treating periodontal disease, Inhibiting composition " However, the composition for controlling plant diseases containing parvitin isolated from the black axes of the present invention as an active ingredient, and the method for controlling plant diseases using the same, have not yet been mentioned.

The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to isolate an active substance from an extract of black shark having high antibacterial activity and identify it as pharbitin, The antimicrobial spectrum was confirmed, and it was found that only the bacterium belonging to Xanthomonas and Ralstonia solanacearum showed high specific activity. Thus, it was confirmed that the extract of black shark and parvitin were effective for the control of plant pathogenic bacteria When developing a bacterial agent to use as a pesticide lead substance, The present invention has been accomplished by anticipating that it will be possible to effectively control diseases caused by various phytopathogenic bacteria, including Rhizoctonia solanaceae.

In order to solve the above-mentioned problems, the present invention provides a plant disease controlling composition comprising pharbitin isolated from the black axis (Pharbitidis Semen) as an active ingredient.

The present invention also provides a method of controlling a plant disease comprising the step of treating an effective amount of the plant disease control composition to a plant part, soil or seed.

The present invention also provides an antimicrobial composition for phytopathogenic fungi which contains pharbitin isolated from the black axis (Pharbitidis Semen) as an active ingredient.

Parvitin according to the present invention has excellent control activity against botanical diseases caused by bacterium of Xanthomonas and Ralstonia solanacearum bacteria without harming to the human body and causing environmental pollution originating from natural products It can be useful for the development of biological pesticides, organic farming materials and environmentally friendly grain production.

1 shows a process of separating and purifying an antibacterial active material from a butanol extract of black shrinkage.
FIG. 2 shows TLC analysis results of the antifungal active compound F211 and the pure compound 1 substance isolated from black shafts.
FIG. 3 is a graph showing the results of a high-resolution-electron atomization-mass spectrometry (HR-ESI-MS) cation (a) and anion mode (a) b) shows the results of the analysis.
Figure 4 shows the chemical structure of parvitin (substance 2-8) and parvitic acid C (substance 1).
FIG. 5 is a photograph (b) showing the effect of controlling the peach bacterial perforation in a detached leaf assay of the parvitin and black-and-white butanol extracts (a) and the results thereof.
Fig. 6 is a photograph (b) of the control effect (a) of the parvitin and the black bean extract of tomato against tomato blight in tomato seedlings and the results.

In order to achieve the object of the present invention, the present invention provides a plant disease controlling composition comprising pharbitin as an active ingredient.

The plant disease control composition according to the present invention is isolated from a natural product, Pharbitidis Semen, and is harmless to the human body, does not cause environmental pollution, and exhibits high controlling activity against various plant diseases.

The parvitin may be a mixture of compounds represented by 2 to 8 among the compounds represented by the following general formula (I), but is not limited thereto.

In the composition according to an embodiment of the present invention, the plant bottle may be a tomato blight, a peach bacterial pore, a citrus peel, a red pepper bloom, or a blight of rice blast fungus, Tomato foot blight by Ralstonia solanacearum , Zanthomonas arboricola P. v. Xanthomonas arboricola pv. Peach Bacterial Hole Disease by Pruni , Zanthomonas aconopodis VIV . Xanthomonas axonopodis pv. citri ), Citrus mellitus disease caused by Xanthomonas euvesicatoria ) or pepper bacillus flecks disease caused by. It may be, but is not limited to, blight of rice blight caused by duckwood ( Xanthomonas oryzae pv. Oryzae ).

The present invention also provides a method of controlling a plant disease comprising the step of treating an effective amount of the plant disease control composition to a plant part, soil or seed.

The composition for controlling plant diseases according to the present invention may further contain substances contained in insecticides or bactericides commonly used in the art in addition to the above-mentioned active ingredients. In addition to the active ingredients, pharmaceutically acceptable solid carriers, Liquid carriers, liquid diluents, liquefied gas diluents, solid diluents, or other suitable adjuvants such as emulsifiers, dispersants or foams. The composition for controlling a mixture of the active ingredient and the above excipient may be formulated into various formulations known in the field of agrochemicals. For formulation, any of the formulation methods commonly used in the field of agrochemicals may be used.

The plant disease controlling composition of the present invention can be formulated into a formulation preferably in the form of a wettable powder, a granule, a powder, an oil, a spray, a film, a capsule or a gel, It is preferably provided as a contact agent.

By treating the composition for control of the present invention, which has been formulated as described above, on a plant or a region requiring control, it is possible to control tomato foot rot disease, peach bacterial perforation, and the like. A preferred method is to apply the control composition to direct contact with plant pathogenic bacteria to control the soil, and it is most preferable to mix the soil or directly spray the crop.

The present invention also provides an antimicrobial composition for phytopathogenic fungi containing pharbitin as an active ingredient.

The parvitin can be isolated from, but is not limited to, seeds of Pharbitidis Semen.

In the antimicrobial composition of the present invention, the plant pathogenic bacterium may be Ralstonia solanacearum or Xanthomonas sp., And preferably the Zanthomonas sp. Ricola P.V. Pruni (Xanthomonas arboricola pv. Pruni ), Zanthomonas axonopodis P. v. Lee Sheets (Xanthomonas axonopodis pv. citri , Xanthomonas < RTI ID = 0.0 > euvesicatoria ) or Jantou Monas Duck as P. v. Duck ash ( Xanthomonas oryzae pv. oryzae strains, but are not limited thereto.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example  One. Black axis  Isolation of Active Substance from Butanol Extract

After 150 g of black seeds were crushed, 2 L of methanol was added, and the mixture was shaken at room temperature for 24 hours. Then, the mixture was filtered through a filter paper, and 2 L of methanol was added to the residue. After shaking for 24 hours, The mixture was concentrated under reduced pressure. 6,171 mg of methanol extract was obtained and dissolved in 500 mL of distilled water. The aqueous solution was then transferred to the follicular phase. Then, 500 mL of ethyl acetate was added to the circulating flask containing the methanol extract, and the residue remaining in the flask was dissolved, and then transferred to a fractionation oven containing the aqueous solution. The fractions containing the aqueous solution and the ethyl acetate solution were shaken for 1 minute, allowed to stand, and then allowed to stand at room temperature until the two layers were separated. The ethyl acetate layer was transferred to an Erlenmeyer flask, and water was removed with anhydrous sodium sulfate. This procedure was repeated twice. The ethyl acetate solution was combined and concentrated under reduced pressure to obtain 1,610 mg of the extract. The aqueous layer was further fractionated twice with butanol and concentrated under reduced pressure to obtain 2,160 mg of butanol extract and 1,700 mg of water extract.

The butanol extract of black beans (2,160 mg) was added to a silica gel column (4 cm x 60 cm, Kiesel gel 60, 150 g, 70-230 mesh) and eluted with an organic solvent of chloroform: methanol: water (14: 6: After the Zanthomonas Arbo Ricola VV. Pruny ( Xanthomonas arboricola pv. investigated the antibacterial activity of the pruni) results, it showed that there is six fractions (F2 ~ F7) activity, the combined fractions 6 to give the F21 fraction (1,571 mg). F21 fraction (1,571 mg) was added to a Sephadex LH-20 column (3 cm x 80 cm, 50 g) and eluted with 100% methanol to obtain four fractions (F211 to F214). F212 (572.7 mg) was repeatedly fractionated by Sephadex LH-20 column chromatography, and the two fractions F211 and F2121 were collected to isolate the active substance F211 (644.5 mg) purely.

The alkaline hydrolysis reaction was carried out to separate the pure compound from the F211 (644.5 mg) compound. To the F211 fraction, 10 ml of 1% aqueous potassium carbonate was added and the mixture was heated at 95 ° C for 2 hours. The mixture was adjusted to pH 4 with 1N hydrochloric acid, and the mixture was partitioned with 10 ml of ethyl acetate. The aqueous layer was concentrated at 60 ° C, ≪ / RTI > The aqueous layer was chromatographed on a Sephadex LH-20 column [3.5 cm x 60 cm, 100 g, 18-111 [mu] m (GE Healthcare, Sweden) and eluted with 100% MeOH to isolate pure Compound 1 (Fig. The obtained compounds F211 and 1 were developed under a solvent condition of chloroform: methanol: water: acetic acid (55:36: 8: 1), observed under ultraviolet rays (UV) at 254 nm and 365 nm, and the TLC development plate was treated with p-anisaldehyde After spraying and heat treatment at 120 캜, F211 showed a light yellow color and pure Compound 1 showed a light blue color (Fig. 2).

Example  2. Determination of the structure of the separated material

As a result of ¹ H-NMR analysis of F211 isolated from black shafts, the chemical structure was estimated to be pharbitin. Parvitine is a hydroxyl fatty acid with multiple sugars and is known as a complex compound consisting of seven substances. It is impossible to separate into a single substance without forming a derivative at F211 separated from the black axis. Therefore, alkali hydrolysis was carried out to find out the exact chemical structure. 470 mg of F211 was dissolved in 1% K ₂ CO ₃ (10 ml) and heated at 90 ° C for two and a half hours. The pH was lowered to 4.0 with 1N HCl and extracted with ethyl acetate (10 ml). After the organic solvent extract was washed with distilled water, the water was completely removed from the organic solvent layer using Na ₂ SO ₄ and concentrated under reduced pressure. Then, a Sephadex LH-20 column (3.5-cm inner diameter and 60-cm length, 100 g , Bead size 18-111 [mu] m, GE Healthcare, Sweden). Methanol / water / acetic acid (55:36: 8: 1, v / v / v / v) was added to the solution, which was then eluted with methanol and then subjected to TLC analysis. ) To obtain pure substance 1 (85.3 mg).

The pure substance 1 was analyzed by high resolution-electron atomization-mass spectrometry (HR-MS) using a Synapt G2 HDMS quadrupole time-of-flight (QTOF) mass spectrometer equipped with an electrospray ion source (Waters, Premier, UK) (Bruker Biospin GmbH, Germany) was used for the determination of ¹ H (ESI-MS) data and was also dissolved in pyridine d ₅ (Cambridge Isotope Laboratories, Inc., Andover, Mass.) Using a Bruker Avance III HD 500 MHz nuclear magnetic resonance analyzer -NMR data was obtained.

HR-ESI-MS analysis showed that the molecular ion [MH] appeared at m / z 1021.4698 in anion mode and the molecular ion [M + Na] ⁺ at m / z 1045.4694 in cation mode (FIG. Also under ¹ H-NMR spectrum exhibited the five anomeric protons (δ 6.26, 5.89, 5.40, 5.21 and 4.87) and three methyls, one primary methyl (δ 0.95), resulting substance 1 summarizes all the data is (Fig. 4), which is consistent with the pharbitic acid C reported by Ono et al. (1990) as shown in Table 1. Therefore, F211 was identified as parvitin as the alkaline degradation product of F211 was identified as parvitic acid C.

Pure compound 1 ¹ H-NMR analysis of the fraction (pharbitic acid C) Carbon Pure compound 1 pharbitic acid C
(Ono et al ., 1990, Chem. Pharm. Bull 38, 1892-1897) Glc-1 4.87 (d, 7.5) 4.92 (d, 7.6) Glc-2 4.32 (dd, 2.5, 5.0) 4.37 (dd, 7.6, 9.0) Glc-3 4.55 (dd, 6.5, 5.5) 4.57 (dd, 9.0, 9.0) Glc-4 3.89 (dd, 3.0, 5.5) 3.94 (dd, 9.0, 9.0) Glc-5 3.90 * 4.01 * Glc-6 3.91 * 4.02 * 4.54 * 4.55 * Glc'-1 5.89 (d, 6.7) 5.94 (d, 7.0) Glc'-2 4.14 * 4.25 * Glc'-3 4.10 (dd, 9.5, 8.5) 4.24 (dd, 9.0, 9.0) Glc'-4 4.02 (dd, 7.5, 7.5) 4.10 (dd, 9.0, 9.0) Glc'-5 3.82 (dd, 3.5, 5.5) 3.88 (ddd, 3.0, 6.0, 9.0) Glc'-6 4.19 (dd, 8.0, 9.5) 4.29 (dd, 6.0, 11.0) 4.39 * 4.46 * Rha-1 5.4 (d, 1.5) 5.40 (d, 1.5) Rha-2 4.53 * 4.54 * Rha-3 4.50 * 4.53 * Rha-4 4.15 (dd, 9.0, 9.0) 4.26 (dd, 9.0, 9.0) Rha-5 4.26 (dq, 5.5, 9.0) 4.36 (dq, 9.0, 6.1) Rha-6 1.63 (d, 6.0) 1.65 (d, 6.1) Rha'-1 6.26 s 6.36 (d, 1.5) Rha'-2 4.75 s 4.72 (dd, 1.5, 3.3) Rha'-3 4.81 (d, 2.0, 9.0) 4.81 (dd, 3.3, 9.6) Rha'-4 4.39 s 4.46 (dq, 9.6, 9.6) Rha'-5 5.09 s 5.09 (dq, 9.6, 6.1) Rha'-6 1.86 (d, 6.0) 1.92 (d, 6.1) Qui-1 5.21 (d, 7.4) 5.27 (d, 7.3) Qui-2 3.98 * 4.09 * Qui-3 3.98 * 4.09 * Qui-4 3.67 (dd, 9.0, 9.0) 3.69 (dd, 9.0, 9.0) Qui-5 3.58 (dd, 6.5, 7.5) 3.59 (dq, 9.0, 6.1) Qui-6 1.52 (d, 6.0) 1.55 (d, 6.1) Ag-2 2.72 (t, 11.7) 2.87 (dd, 15.0, 5.5) 2.87 (d, 14.4) 2.91 (dd, 15.0, 8.0) Ag-3 4.38 * 4.45 * Ag-11 3.86 * 3.90 * Ag-14 0.95 (t, 6.9) 0.97 (t, 7.0)

Example  3. Of parvitin (F211)  For various plant hospital bacteria Antimicrobial activity

The F211 fraction and butanol extract obtained from Example 2, identified as parvitin, were tested for their antibacterial activity against various phytopathogenic bacteria. Gram-negative and Gram-positive strains were serially diluted to a maximum of 500 ㎍ / ㎖ sequentially, and the minimum growth inhibitory concentration (MIC) was determined by 96-well microplate bioassay to 6.25 ㎍ / Respectively. Streptomycin sulfate was used as a positive control and diluted to a maximum of 6.25 μg / ㎖ sequentially from 100 ㎍ / ㎖ up to 2 times. 14 plants suspected of being stored at -80 ° C in a 20% glycerol solution were inoculated into a solid medium of tryptic soy agar (TSA) and cultured in a stationary condition. 10 ml of a tryptic soy broth (TSB) was placed in a test tube, the inlet was closed with sterilization, sterilized, and one of the colonies of the strains which had been subjected to the stationary culture was scraped with a stripper and placed in a sterilized liquid medium. Respectively. The liquid cultured plant pathogenic bacteria were adjusted to an OD value of 0.1 using a UV spectrophotometer, diluted with sterilized water, and inoculated into a 96-well plate at 10 ⁶ CFU / ml. Butanol extract and Streptomycin sulfate, which is a positive control, were treated with the cultures of plant pathogenic bacteria. After incubation for 1 ~ 3 days at the optimal temperature condition, the minimum growth inhibitory concentration (MIC) was investigated. Growth of plant pathogenic bacteria was measured with an absorbance of 595 nm in a microplate reader and was performed with 2 replicates per treatment.

Optimal Growth Conditions of 13 Various Plant Pathogens Used for Antibacterial Activity Assay Plant pathogen Culture conditions badge Temperature (℃) Day of cultivation Ashibo Lovers Abe My Subtotal Catlia
( Acidovorax avenae subsp. cattlyae ) TSA, TSB 30 One Ashido Lovers Konjasi
( Acidovorax conjugate ) Agrobacterium tumefaciens
( Agrobacterium tumefaciens ) Burkholdeldia gluma
( Burkholderia glumae ) Pectobacterium carotovora subspecies carotovora
( Pectobacterium carotovora subsp. carotovora ) Peptobacterium chrysanthemi
( Pectobacterium chrysanthemi ) Pseudomonas sylin G. P. V. Actini
( Pseudomonas syringae pv. actinidae ) 25 Pseudomonas sylin G. P. V. Lacrimans
( Pseudomonas syringae pv. lachrymans ) 30 2 Ralstonia Solana Seorum
( Ralstonia solanacearum ) 28 Zanthomonas Arvor Ricola PV. Pruny
( Xanthomonas arboricola pv. pruni ) Zanthomonas evonopodispev. Citi
( Xanthomonas axonopodis pv. citri ) Jantou Monas Yubecikatoria
( Xanthomonas euvesicatoria ) Zanthomonas duck asp. V. Duck
( Xanthomonas oryzae pv. oryzae ) 30 3 Clavi Better microorganisms
Clavibacter michiganensis subsp. michiganensis ) 30 2

The minimum inhibitory concentration (MIC) for the various plant pathogenic pathogens of parvitin and black bean extracts is shown in Table 3 below. As with the black bean extract, parvitin showed antibacterial activity only against the bacteria of the genus Zanthomonas and Ralstonia solanaceae among the target plant pathogenic bacteria. Particularly, it has been reported that xanthomonas oryzae pv. oryzae) and tomato was a minimum inhibitory concentration of 31.25 ㎍ / ㎖ for LAL Stony Oh Solana Seah column (R. solanacearum), causing the foot blight, bacterial pathogens jeommunuibyeong (Xanthomonas euvesicatoria), peach bacterial hole disease causing bacteria (Xanthomonas arboricola pv. The minimum growth inhibitory concentration was 102 ㎍ / ㎖ for citrus juvenile ( Xanthomonas axonopodis pv. citri ).

The minimum growth inhibition concentration (MIC) value of 14 plant pathogenic pathogens of parvitin and black bean extracts Plant pathogen Minimum growth inhibitory concentration (MIC, 占 퐂 / ml) Butanol
extract Pharbitin Streptomycin sulfate Ashibo Lovers Abe My Subtotal Catlia
( Acidovorax avenae subsp. cattlyae ) - - - Ashido Lovers Konjasi
( Acidovorax conjugate ) - - 6.25 Agrobacterium tumefaciens
( Agrobacterium tumefaciens ) - - 12.5 Burkholdeldia gluma
( Burkholderia glumae ) - - 12.5 Clavi Better microorganisms
Clavibacter michiganensis subsp. michiganensis ) - - 25 Pectobacterium carotovora subspecies carotovora
( Pectobacterium carotovora subsp. carotovora ) - - 6.25 Peptobacterium chrysanthemi
( Pectobacterium chrysanthemi ) - - - Pseudomonas sylin G. P. V. Actini
( Pseudomonas syringae pv. actinidae ) - - 12.5 Pseudomonas sylin G. P. V. Lacrimans
( Pseudomonas syringae pv. lachrymans ) - - 12.5 Ralstonia Solana Seorum
( Ralstonia solanacearum ) 125 31.25 6.25 Zanthomonas Arvor Ricola PV. Pruny
( Xanthomonas arboricola pv. pruni ) 125 62.5 12.5 Zanthomonas evonopodispev. Citi
( Xanthomonas axonopodis pv. citri ) 500 125 50 Jantou Monas Yubecikatoria
( Xanthomonas euvesicatoria ) 125 62.5 12.5 Zanthomonas duck asp. V. Duck
( Xanthomonas oryzae pv. oryzae ) 125 31.25 6.25

-: No activity

Example  4. Peach leaves Parvitin Bacterial hole disease  Control active black

In order to investigate the effect of the bacterial shot holes on the bacterial shot holes, 30 ℓ pots mixed with 1: 1 ratio of soil and sand were exposed to 3 - year - old peach trees of Aozora Musume at 28-32 ℃, % The leaves (6-9 cm) of the young branches of the peach grown in a greenhouse were washed three times with sterile distilled water, placed on a sterilized paper towel and dried. The extracts of parvitin and butanol were dissolved in methanol to a concentration of 50, 25 and 12.5 mg / ml, and the respective stock solutions were diluted 100 times with distilled water containing 250 μg / ml of Tween-20, Ml. ≪ / RTI > Tween-20 (250 占 퐂 / ml) solution containing 1% methanol was used as the control, and Tween-20 (250 占 퐂 / ml) containing 200 占 퐂 / ml of streptomycin sulfate was used as a positive control. Peach leaves were soaked in parvitin and butanol extract, Tween-20, and streptomycin sulfate solution for 10 seconds, and placed on a 0.5% agar agar. After the passage of time, the cells were incubated in a test tube at 28 ° C and 150 rpm for 36-48 hours Shake cultivated peach Bacterial pest pathogenic agent Bacterium Monas Arbo Ricola PB. Rooney program taking the (X. arboricola pv. Pruni) a UV spectrophotometer 6 wound the OD value with the meter to a peach bacterial germs hole buried on the forceps according to 0.1 (10 ⁸ CFU / ㎖) peach leaf pathogens Respectively. And sealed for 8 hours under light-free conditions at 28 DEG C for 16 hours, and the change in leaves for 5 days was recorded. Two leaves were used in one petri dish per treatment and repeated twice. The incidence rate was 50% when six out of twelve pathogens were inoculated, and disease incidence and control were calculated using the following formula.

% Disease incidence (DI) = 100 × [Number of affected cases / Number of all injuries]

% Disease control = 100 - [100 × incidence of treatment / incidence of non-treatment]

As a result, as shown in Fig. 5, the parvitin and butanol extracts decreased the incidence of peach bacterial pores in a concentration-dependent manner, while parvitin decreased the peach bacterial pore size at 50, 70.8 , 87.5%, and the butanol layer also inhibited 29.2, 54.2 and 75.0% at the concentrations of 125, 250 and 500 ㎍ / ㎖. Therefore, the anti-bacterial substance parvitine isolated from the black axis of the present invention is an anti-bacterial substance, It was confirmed that X. arboricola pv. Pruni had excellent antibacterial activity.

Example  5. Tomatoes From seedlings Parvitin  Activity control of bacterial foot blight disease control

In order to investigate the antiseptic activity of parvitin and black bean extracts on the tomato root rot caused by tomatoes, the seeds of Suhwang (Palm Han - Nong) tomato juice (Soju Cup) filled with commercial horticultural soil And after 24 days 4-5 leaves of tomato were transferred to a drinking cup (diameter 7 cm). The butanol extracts were dissolved in methanol at concentrations of 50, 25 and 12.5 mg / ml with methanol at the concentrations of 200, 100 and 50 mg / ml, and the respective stock solutions were mixed with distilled water containing 250 μg / ml of Tween- And the butanol extract was diluted to the levels of 2000, 1000 and 500 / / ml. Twenty milliliters of the parvitin and butanol extracts prepared in the tomato that had passed 24 hours after the passage was evenly distributed in the soil . The OD value was adjusted to 0.1 (10 < ⁸ > CFU / ml) using a UV spectrophotometer, and the bacterial counts of parvitin and paclitaxel Six hours after the butanol extract treatment, 20 ml of the Pseudomonas aeruginosa suspension was evenly distributed in the soil to inoculate the pathogens. Tween-20 (250 占 퐂 / ml) solution containing 1% methanol was used as the control, and Tween-20 (250 占 퐂 / ml) containing 200 占 퐂 / ml of streptomycin sulfate was used as a positive control. Five tomato plants were used for each treatment, and they were repeated three times. Tomato plants were observed at 30 ± 2 ° C and relative humidity of 70-80%.

As shown in FIG. 6, tomatoes treated with parvitin 125, 250 and 500 / / ml after 7 days of treatment with parvitine and butanol extracts were controlled to 79.5, 97.4 and 100% The treated tomatoes treated with 500, 1000 and 2000 ㎍ / ㎖ of butanol extract were also controlled to 76.9, 100, 100%. However, on the 2nd day from the treatment with 2000 ㎍ / ㎖ of butanol extract, the leaf was developed as a yellow lesion. After 14 days of treatment with parvitine and butanol extracts, the control effect of foot wilt disease decreased compared with that after 7 days. Therefore, it was confirmed that the antibiotic parvitin isolated from the black axis of the present invention has excellent antibacterial activity against R. solanacearum , a causative agent of tomato foot rot.

Claims (8)

A composition for controlling plant diseases comprising pharbitin as an active ingredient. The plant disease controlling composition according to claim 1, wherein the parvitin is isolated from a seed of Pharbitidis Semen. [3] The plant disease control composition according to claim 1, wherein the plant disease is at least one selected from the group consisting of peach bacterial pericarditis, citrus peptic ulcer disease, red pepper bloom spot blight disease and rice blight blight disease. A method for controlling a plant disease comprising treating a plant part, soil or seed with an effective amount of the plant disease controlling composition according to any one of claims 1 to 3. An antimicrobial composition for phytopathogenic fungi containing pharbitin as an active ingredient. 6. The antimicrobial composition according to claim 5, wherein the parvitin is isolated from a seed of Pharbitidis Semen. The antimicrobial composition according to claim 5, wherein the plant pathogenic bacterium is a strain of Ralstonia solanacearum or Xanthomonas sp. 8. The method according to claim 7, wherein the Zanthomonas sp. Strain is a Zanthomonas arboricola var. Rooney program (Xanthomonas arboricola pv. pruni ), Zanthomonas aconopodis P v. Li sheet (Xanthomonas axonopodis pv. Citri), Pseudomonas janto Juve Brassica thoria (Xanthomonas euvesicatoria) or Pseudomonas janto duck material P V. (Xanthomonas oryzae pv. Oryzae ). ≪ / RTI >

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