KR100422883B1 - Fungicide composition for Control of Phytophthora Blight of Pepper containing Antifungal Compounds Phenylacetic Acid and Sodium phenylacetate - Google Patents

Abstract

The present invention relates to an environmentally friendly natural plant disease control fungicide composition that can be naturally decomposed by other microorganisms in nature as well as to control plant diseases by containing natural substances derived from microorganisms, natural substances derived from actinomycetes Phosphorus antibacterial agent of the present invention which contains the phosphorus antibacterial substance phenylacetic acid and sodium phenylacetate as an active ingredient, Phytophthora capsici , choletotricum obiculus It is effective in controlling plant diseases caused by major phytopathogens such as Colletotrichum orbiculare , Rhizoctonia solani , and Pythium ultimum .

Description

Fungicide composition for Control of Phytophthora Blight of Pepper containing Antifungal Compounds Phenylacetic Acid and Sodium phenylacetate}

The present invention relates to a bactericidal composition for controlling red pepper blight, which comprises the antimicrobial substance phenylacetic acid and sodium phenylacetate as active ingredients, and more specifically, Phytophthora capsici , Colletotricum obiculle Orbiculare ), Rhizoctonia solani ( Rhizoctonia solani ), Pythium ultimum ( Fythium ultimum ) and the like fungi (fungal) component effective for the treatment of fungal diseases and a fungicide composition containing the same as an active ingredient.

Modern crop production systems continue to evolve to protect crops from the constant threats of harmful organisms that hinder productivity gains. In modern agriculture, efficient control of plant diseases is not conceivable except for fungicides. Reliance on fungicides, especially in developed countries, is much higher, with 39% of the world's disinfectant production used in Western Europe, 28% in the Far East and 12% in the United States, with only 21% used in the rest of the world. .

Control of plant diseases by fungicides Bordeaux Mixture, developed by Alexis Millardet in the 1850s, was used to control vine fungi caused by Plasmopara viticola . Since a large number of organic fungicides were developed in the mid-1900s and started to control various plant diseases, more than 200 fungicides with 113 active ingredients have been used.

In general, pepper blight is economically very important in our country and has become a big factor in pepper production. In order to control this, cultivation of resistant varieties, rotation, biological control using antagonists, etc. have been carried out in various ways, but until now, the most effective control method is known to be chemical control by disinfectant spraying.

Metalaxyl, fosetyl-Al, oxadixyl, propamocarb, copper oxychloride, chlorothalonil, dithianon, etc. Currently, it is used as a fungicide for controlling red pepper disease in Korea, and metalaccil-caperoxychloride, metalaccil-dithianon, oxadisil-chlorotalonyl, and the like are also used as a mixing agent. However, despite the excellent efficacy of these drugs, the compound having a new structure, or a new compound has been recently confirmed the interest due to the emergence of drug-resistant strains.

In order to control the pepper disease, approximately 1,000 M / T (1993) of organic synthetic fungicide is sprayed on 83,000 red pepper plantations, causing severe adverse effects on the environment caused by pesticides and the possibility of drug resistant strains. Doing. Therefore, from this point of view, it can be said that there is an urgent need for development of an eco-friendly fungicide that does not destroy the environment as well as low toxicity for the control of the pepper blight.

Phenylacetic acid, which is enzymatically derived from the amino acid phenylalanine, is known to act as a natural growth regulator that promotes the growth of plants such as corn, barley, tobacco, or tomatoes. It has also been reported that the fungus Penicillium chrysogenum uses phenylacetic acid as a precursor when biosynthesizing the antibiotic penicillin G. The pharmacological action of phenylacetic acid, which has been known to date, has been shown to have an inhibitory effect on some nematodes that actually affect wood, but phenylacetic acid and sodium phenylacetate itself have excellent effects as a pollution-free natural fungicide. Nothing has been reported.

The present inventors have been conducting researches to develop antifungal antibiotics from soil microorganisms since 1990 in the Department of Agricultural Biology, Korea University, and have discovered a number of antagonistic fungi that produce antifungal antibiotics showing excellent antimicrobial activity against phytopathogenic fungi. It was. As a result of screening the efficacy of several antimicrobial substances separated and purified as a single component from the antagonist, it was confirmed that two of the antimicrobial substances have an excellent antimicrobial activity against phytopathogenic fungi, and the molecular structure of the antimicrobial substances was identified. As a result, it was confirmed that the phenylacetic acid and sodium phenylacetate to complete the present invention.

Accordingly, it is an object of the present invention to provide a fungicide composition for controlling plant diseases using phenyl acetic acid and sodium phenyl acetate. Another object of the present invention is to provide a use of the phenyl acetic acid and sodium phenyl acetate as an antifungal agent.

The purpose of the present invention is to contain natural substances in plant fungicides for controlling plant diseases, specifically, by containing antimicrobial substances phenylacetic acid and sodium phenyl acetate derived from actinomycetes as an active ingredient, as well as other effects in the natural environment. Manufacture of biodegradable biodegradable fungicides that are naturally decomposed by microorganisms, and compare the inhibitory activity against phytopathogenic fungi, the antibacterial activity against red pepper bacillus and the control against late blight in existing greenhouses Achieved by evaluation.

1 is a graph showing the germination rate and mycelial growth rate of the red pepper bacterium according to the concentration of the antimicrobial material of the present invention.

Figure 2 is a graph showing the results of comparing the control effect of the present invention antibacterial substance against red pepper blight in pot experiments of greenhouse conditions with metalluxyl (metalaxyl) and fosetyl aluminum (phosphateyl-Al), phosphoric acid as a commercial fungicide .

The present invention comprises the steps of selecting the actinomycetes strain to isolate and purify the antimicrobial active substance phenylacetic acid (phenylacetic acid) and sodium phenylacetate; Pna capsici , including Magnaporthe grisea , Alternaria mali , Colletotrichum orbuculare , Cylindrocarpon destructans ), Fusarium moniliforme , Fusarium oxysporum f.sp. Phenylacetic acid and sodium phenyl for several plant fungal pathogens such as Fusarium oxysporum f.sp.cucumerinum , Pythium ultimum and Rhizoctonia solani Examining the inhibitory activity of acetate (sodium phenylacetate); Measuring the antimicrobial activity of phenylacetic acid and sodium phenylacetate against red pepper bacterium; This study consists of comparing the effects of phenylacetic acid and sodium phenylacetate and the late blight control agents metalaccil, pocetyl aluminum and phosphoric acid on greenhouse seedlings.

When the phenylacetic acid isolated and purified in the present invention has been used as an antibiotic until now, the activity as an nematicide has been reported, but the effect as a fungicide for controlling pollution of natural plant diseases has not been disclosed as in the present invention. Moreover, sodium phenylacetate has not been reported for any pharmacological action.

The present inventors conducted an inhibition test for phytopathogenic fungi in order to confirm the control effect of the antimicrobial substance phenylacetic acid and sodium phenylacetate represented by the following formula (I), the control group that is a conventional organic synthetic fungicide Compared with the results, it was confirmed that there was an equivalent level of control effect. In particular, the antimicrobial substances phenylacetic acid and sodium phenylacetate used in the present invention are not only the first to be found in actinomycetes of Streptomyces genus, but also to choltotricum obiuleur ( Phytophthora capsici ) Colletotrichum orbiculare ), Rhizoctonia solani , and Pythium ultimum also have specific antimicrobial activity and mechanism of action.

(In the above formula, R = H in phenylacetic acid and R = Na in sodium phenylacetate.)

The antibiotic of the present invention may be used alone or in combination with a filler commonly used in the art to which the present invention pertains. That is, the composition which uses the antibiotic of this invention as an active ingredient can be manufactured and used.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Isolation and Purification of Antimicrobial Phenylacetic Acid and Sodium Phenylacetate

The present inventors separated about 100 actinomycetes from soil samples in various forests and coastal areas of Korea, and tested the control effect on agar plate and plants under greenhouse conditions. The antimicrobial phenylacetic acid (phenylacetic acid) and sodium phenylacetate (sodium phenylacetate) were isolated and purified, and the structure thereof was determined.

Actinomycetes strain S5-55 identified as Streptomyces humidus was cultured in starch broth for 14 days at 28 ° C, and then the filtrate was separated from actinomycetes mycelium by centrifugation. The separated culture filtrate was extracted with butanol, and the extract was concentrated under reduced pressure, followed by C18 reversed phase chromatography. Each fraction was developed by mixing water and methanol in a predetermined ratio, and the active fractions were collected and preparative TLC (Thin-Layer Chromatography) was performed. The active fraction shown on the TLC was collected using a UV detector, extracted with methanol, and subjected to gel filtration chromatography at a flow rate of 0.05 mL / min using Sepadex-LH 20 resin. C18 reverse-phase HPLC (High-Performance Liquid Chromatography) was performed by collecting the active fractions of the two groups from the fractions obtained through gel filtration chromatography using preparative conditions using a flow rate of 2 ml / min, UV detection 210 nm, and a solvent 10% to 100% aqueous acetonitrile. Antibacterial phenylacetic acid and sodium phenylacetate were isolated and purified.

Example 2: Inhibition test of various phytopathogenic fungi of antimicrobial substance phenylacetic acid and sodium phenylacetate

P. capsici , a bacteriophage bacterium, Magnaporthe grisea , Alternaria mali , Colletotrichum orbuculare , and cylindrocapon destruct Cylindrocarpon destructans , Fusarium moniliforme , Fusarium oxysporum f.sp. Phytopathogenic fungi , such as Fusarium oxysporum f.sp.cucumerinum , Pythium ultimum , and Rhizoctonia solani , were cultured in potato dextrose agar medium. .

The phenylacetic acid and sodium phenylacetate separated and purified in Example 1 were mixed with 1% potato agar liquid medium in a micro-well plate and contained in a series of concentrations (1 to 1,000 ㎍ / ml). Afterwards, each fungal spore or citron suspension was placed in each well at a concentration of 10 ⁴ spores / ml. The inoculated microwell plate was incubated at 28 ° C. for 2 to 4 days, and the lowest concentration among the fungi without mycelial growth was evaluated as minimum inhibitory concentration (MIC), and the results are shown in the following table. 1 is shown.

Minimum Inhibitory Concentration (MIC) of Phenylacetic Acid and Sodium Phenyl Acetate on Various Plant Pathogens Phytopathogenic fungi Minimum Inhibition Concentration (μg / mL) Phenylacetic acid Sodium Phenyl Acetate Alternaria Malicoletotricum Obiculure Cylinder Locuson Dispostanum Pulinium Monoliform Fussium Oxyporum 500 100 500 1,000 1,000 100 50 1050 1,000501,0001,0001,0001,000501050

The phenylacetic acid showed high levels of antimicrobial activity against P. capsici , Pythium ultimum and R. solani , , Sodium phenyl acetate also showed antimicrobial activity against the three fungi and Colletotrichum orbiicular ( colletotrichum orbiculare ). However, it did not show antimicrobial activity against the genus Alternaria, Cylindrocarpon destructans, and Fuzarium.

Example 2 Antimicrobial Activity of the Antibacterial Substance Phenyl Acetate and Sodium Phenyl Acetate

Pepper disease is a phytopathogenic bacterium that forms a yuji in the actual pepper packaging, causing a sudden spread of the disease, and the first half of the yuji soon leads to the development of the disease. For this reason, it is important to check the antimicrobial activity against the juniper in the control of the red pepper disease. Antimicrobial activity of the antimicrobial substance phenylacetic acid and sodium phenylacetate against red pepper bacterium strains was confirmed under an optical microscope.

Specifically, phenylacetic acid and sodium phenylacetate diluted with a series of concentrations (1-500 μg / ml) were prepared by making a yeast suspension (2 × 10 ⁷ sachets / ml) of the caybean germ bacterium, and meta as a control agent. After 4 hours of mixing in a tube containing lacxyl, pocetyl aluminum, and phosphoric acid, the presence or absence of germination of germs was observed under an optical microscope. In addition, the growth inhibition of the hyphae produced from germinated strains was also observed.

As a result, as shown in Fig. 1, it was confirmed that phenylacetic acid and sodium phenylacetate completely inhibited germinal germination at a concentration of 50 µg / ml, and 50 µg / l of phenylacetic acid for mycelial growth. At least 50% inhibition was observed at the concentration of ml. However, sodium phenyl acetate was found to be somewhat inferior to the mycelial growth inhibitory effect compared to phenylacetic acid.

Example 3 Comparison of the Plague Control Effects of Phenylacetic Acid and Sodium Phenyl Acetate in the Greenhouse Seedlings with Metalaccil, Pocetyl Aluminum, and Phosphoric Acid

Phenylacetic acid, sodium phenylacetate, metalaxyl, pocetyl aluminum, and phosphoric acid were treated at concentrations one day before inoculation of the bacterium to reduce the disease severity in each treatment and untreated treatment. Evaluated. In other words, the degree of disease damage for evaluating the control effect of red pepper disease was evaluated in units of 0 to 5 as follows according to the degree of disease damage in the red pepper seedlings treated based on when the untreated red pepper seedlings were fully ill. Grade 0 is visible when no symptoms of visible disease are seen, and grade 5 (0: no outbreak, 1: mildly wilted and necrotic lesions) when pepper seedlings are fully diseased and die. , 2: 30-50% of the whole plant, 3: 50-80% of the whole plant, 4: 80-90% of the whole plant, 5: the death of the plant).

As a result, brown plaques appeared on the stems of untreated red pepper plants from day 4 after inoculation, but no symptom was observed in the pepper plants treated with various control drugs, phenylacetic acid and sodium phenylacetate. Did. At 6 days after inoculation, lesions were also observed in red pepper plants of 10 μg / ml and 100 μg / ml phenylacetic acid and sodium phenylacetate. Some withering symptoms were also observed in red pepper plants treated with metalaccil 10 μg / ml. Pepper plants that were not treated with the drug were completely killed at 9 days after inoculation, but the pepper plants treated with the antimicrobial phenylacetic acid 1,000μg / ml were inoculated with the back pathogen (Fig. 2 A) and soil irrigation. In all cases (Fig. 2B), the incidence of late blight was markedly reduced, and sodium phenylacetate was also reduced at 1,000 μg / ml of drug treatment (Fig. 2A). There was no late blight on the treatment of metalaccil over 100μg / ml.

In view of the test results as described above, the phenylacetic acid used in the present invention has a strong antibacterial activity and antimicrobial effect, although somewhat inferior to the antimicrobial effect of the organic synthetic fungicide component, and is naturally derived from a natural substance. It can be said that it is a substance that can be easily degraded by microorganisms in the environment.

As described above, the present invention includes the antimicrobial substance phenylacetic acid and sodium phenylacetate, which are natural substances derived from actinomycetes, as active ingredients, while having a high control effect at the level of organic synthetic pesticides in a natural environment. It is effective to provide biodegradable biodegradable biocidal composition of microorganisms, Phytophthora capsici , Colletotrichum orbiculare , Rhizoctonia solani , Pythium ultimum ( Pythium ultimum ) is a very useful invention in the plant disease control industry because it has an excellent effect on the control of plant diseases caused by major phytopathogens .

Claims (4)

Phytophthora capsici , Colletotrichum orbiculare , characterized by containing the antibacterial phenylacetic acid or sodium phenylacetate as an active ingredient A fungicide composition for controlling red pepper disease caused by a pathogen of any one of Rhizoctonia solani and Pythium ultimum . delete delete delete