KR101614426B1 - Composition for controlling plant disease containing Galla chinensis extract as effective component and uses thereof - Google Patents

Abstract

The present invention is a composition for controlling plant diseases comprising the nut gall extract as an active ingredient, and relates to the use thereof, and more particularly, gallnut (Galla chinensis extract as an active ingredient, and a method for controlling a plant disease comprising the step of treating the plant or seed with a composition for controlling the plant disease.

Description

TECHNICAL FIELD The present invention relates to a composition for controlling plant diseases, which contains an extract of Opuntia ficus as an active ingredient, and a use thereof.

The present invention is a composition for controlling plant diseases comprising the nut gall extract as an active ingredient, and relates to the use thereof, and more particularly, gallnut (Galla chinensis extract as an active ingredient, and a method for controlling a plant disease comprising the step of treating the plant or seed with a composition for controlling the plant disease.

Acidovorax , a pathogenic rot fungus of crops and crops, avenae subsp. citrulli ) was first reported in the US watermelon plantation in 1965 and caused major damage in the United States from 1989 to 1995. This disease is mainly caused by seed infection, and it is expected that the damage caused by contaminated seeds will occur.

Previously, the bacteria of the genus Acidovorax were Pseudomonas , . However, according to the development of taxonomy, it was newly classified by Willems et al. In 1992. It is classified into 8 species and 3 subspecies by pathogenicity, physiological biochemical characteristics, DNA base composition and 16S rRNA sequence analysis. 4 kinds of these three sub-species (A. anthurii, A. avenae subsp. Avenae, A. avenae subsp. Cattleyae, A. avenae subsp. Citrulli, A. konjaci, A. valerianellae) is a plant pathogenic bacterium four kinds (A. defluvii , A. delafieldii , A. facilis , A. temperans ) are non-pathogenic bacteria.

In particular, Lux ah ah you know a plant pathogenic bacterium walk Bene Bene subspecies (A. avenae subsp. Avenae) and you know walking Rocks Ah Venetian subspecies sheet rulri (A. avenae subsp. Citrulli) are distributed worldwide each millet and watermelon Which is causing great damage. Korea has been a two-kind three subspecies (A. avenae subsp. Avenae, A. avenae subsp. Cattleyae, A. avenae subsp. Citrulli, A. konjaci) reports, etc. millet, corn, Phalaenopsis, watermelon, sesame seeds, lettuce and know walk A. avenae subsp. Avenae and A. avenae subsp. Citrulli , Only a few taxonomic characteristics of bacteria are known.

In general, Acidovorax The genus Bacillus is an important seed of the millet or watermelon. The infected seed is the primary infectious agent. Since it is difficult to control even chemical synthetic pesticides as in other bacterial diseases, it is known to be an effective control method to remove infectious seeds or seedlings by early testing to remove infectious seeds or seedlings.

Bacterial fruit blotch (BFB) is predominantly infected by seeds. Poultry and vegetable seeds are known to survive for two years, and foreign countries are reported to survive for 40 years. Due to the grafting operation, it can be transmitted to a wide range of secondary infections, and even secondary infections can be secondary infections. This disease often occurs in humid conditions of 20 ° C or higher, especially in relatively hot and humid environments of 30 to 35 ° C. Since watermelon fruit rot disease is a bacterial disease, the pathogen has a strong pathogenicity against various foliage and vegetables, so care should be taken not to spread the damage to watermelon and melon as well as other foliage and vegetables.

In Japan, care was taken to prevent pathogens from invading Japan through the Harmful Animal and Plant Quarantine Act, which requires cultivated area inspections for importer countries based on the Enforcement Regulation of the Plant Protection Act. However, watermelons were reported to occur in Akita Prefecture, Yamagata Prefecture, Tottori Prefecture, Nagano Prefecture, and Tokushima Prefecture in 1998, 2001 and 2009, respectively. In Korea, fruit rot caused by watermelon occurred intermittently after it was first reported in Gochang, Jeonbuk in 1991, and it has occurred in 2005 ~ 2006 and 2009 in watermelon packaging and nursery of Gangnam area . This disease is a bacterial disease. Domestic damage is the biggest problem in watermelon, but it has a strong pathogenicity to various kinds of foil and vegetables such as cucumber, pak, melon and melon. Measures need to be established.

Although synthetic pesticides are able to secure stable food all over the world, many misuse / abuse in the control of crops and health hygiene pests cause various problems such as toxicity and pollution of the manure. As a result, the use of synthetic pesticides is being reduced internationally, and OECD member countries are advised to cut their raw material production by 40% by 2013 by 2004. In Japan, agrochemicals based on gasamycin and copper hydrate are registered to prevent fruit rot, but there are no pesticides registered in Korea until now, and it is industrialized as environment friendly organic material which can control environmentally friendly fruit rot disease There is no product.

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Korean Patent No. 1008318 discloses a composition for controlling plant diseases, which contains a white algae extract as an active ingredient, and Korean Patent No. 1104863 discloses a composition for controlling plant diseases, which contains an extract as an active ingredient However, it is different from the composition for controlling plant diseases, which contains the extract of Opuntia ficus-indica according to the present invention as an active ingredient.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a method for controlling botanical disease caused by natural products, which minimizes toxicity to human body while minimizing environmental damage, Rot know the cause of (bacterial fruit blotch) and completed the present invention by confirming that the excellent antibacterial activity against the foot flux Oh Venetian subspecies sheet rulri (Acidovorax avenae subsp. citrulli).

In order to solve the above problems, the present invention gallnut (Galla chinensis ) extract as an active ingredient.

In addition, the present invention provides a method for controlling a plant disease comprising the step of treating the plant disease controlling composition with a plant or seed.

According to the present invention, a composition containing an extract of Rhododendron japonica as an active ingredient can effectively control a bacterial fruit blotch in an environmentally friendly manner, so that the export of a bacterial pathogen (fruit-rot disease) , Which can lead to increased farm income, which will contribute to sustainable agriculture.

Figure 1 shows the growth inhibition of the pathogen after the inoculation of A. avenae subsp. Citrulli pathogen in the medium.
Fig. 2 is a growth inhibition curve showing the degree of growth of pathogens after inoculation of A. avenae subsp. Citrulli pathogen into the medium containing the contrast agent Kasugamycin.
FIG. 3 is a growth inhibition curve showing the growth of pathogens after inoculation of A. avenae subsp. Citrulli pathogen into a medium containing 1,000 ppm of Galla chinensis extract.
Fig. 4 shows a liquid formulation BFB-3 having a high antibacterial activity containing a rhizome extract.
FIG. 5 is a photograph showing bacterial fruit rot of watermelon after 30 days of foliar treatment of A. avenae subsp. Citrulli pathogens in Watermelon seedlings.

According to an aspect of the invention, the invention is gallnut (Galla chinensis ) extract as an active ingredient.

In the composition for controlling plant diseases according to the present invention, the plant disease may be fruit and vegetable disease of a plant and a crop, and the fruit and vegetable fruit rot of the plant may be a bacterial fruit blotch, It does not. The bacterial watermelon fruit rot (Bacterial fruit blotch) organisms are know walk flux (Acidovorax) species, and preferably walk flux Oh Venetian subspecies sheet rulri know of a (Acidovorax avenae subsp. Citrulli).

Further, in for plant disease controlling composition of the present invention, the nut gall (Galla chinensis ) extract can be prepared by extraction with an organic solvent. As the organic solvent for extraction in the present invention, a lower alcohol such as methanol, ethanol, propanol, butanol or an aqueous solution thereof, hexane, ethyl acetate, acetone, methylene chloride or the like may be used alone or in combination. For example, the rhizome extract may be prepared by extracting crushed dwarf with 1 to 3 times of 90 to 100% (v / v) methanol, and then concentrating the mixture at a reduced pressure at 35 to 45 ° C. Preferably, (v / v) methanol and then concentrated under reduced pressure at 40 ° C. The solvent used in the fractionation process may be n-hexane, chloroform (CHCl ₃ ), ethyl acetate (EtOAc) or n-butanol (BuOH ), But the present invention is not limited thereto.

In addition, the composition for controlling plant diseases of the present invention may further contain substances contained in commonly used insecticides or bactericides. In addition to the above-mentioned extracts of Opuntia series, there may be mentioned pesticides such as solid carriers, liquid carriers, liquid diluents, A liquid diluent, a liquid diluent, a solid diluent, or other suitable adjuvant such as an emulsifying agent, a dispersing agent or a foaming agent. The composition for controlling plant diseases mixed with the extract of Opium phytotifinus and the above excipients 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 can 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, But are not limited thereto.

For example, a liquid formulation can be prepared by mixing a variety of supplementary ingredients with a crude extract to lower the susceptibility to weakness and increase the effect of the active ingredient. The supplementary ingredients may further include glycerol, ethanol, and water. Specifically, the liquid formulation may be prepared by mixing Galla chinensis extract, glycerol, ethanol and water in a volume ratio of 30: 40: 4 to 6: 30: 40: 20 to 30, more specifically, Galla chinensis extract, glycerol, ethanol and water at a ratio of 35: 5: 30 to 40: 20 to 30 by volume. The formulations prepared with the materials and the blending ratio has excellent advantages antibacterial activity and physical stability for the know walk flux Oh Venetian subspecies sheet rulri (Acidovorax avenae subsp. Citrulli) compared to the formulations prepared with the addition of other auxiliary materials or other ratio .

In addition, in the composition for controlling plant diseases according to the present invention, it is possible to dilute the rhizome extract to a concentration of 500 to 10,000 ppm, preferably 1,000 to 5,000 ppm in order to effectively control the plant disease, The degree of growth, the cultivation environment, the degree of plant disease incidence, and the like.

The present invention also provides a method for controlling a plant disease comprising the step of treating the plant with a plant disease controlling composition. The plant disease is as described above.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1: Park and crops In fruit rot  Selection of natural substances derived from plant-active antibacterial plants

Plant-know to target the material that exhibits the activity of bacterial pathogens from natural substances foot flux Oh Venetian subspecies sheet rulri (A. avenae subsp citrulli.) Flux of hospital bacteria-only medium AacSM (know foot ah Venetian selective medium: sodium phosphate 2.5 g 0.5 g of potassium phosphate, 2.0 g of ammonium sulfate, 0.029 g of magnesium sulfate, 0.067 g of calcium chloride, 0.025 g of sodium molybdate, 0.010 g of yeast extract, 10 g of ammonium adipate, 0.0125 g of BTB solution (bromothymol blue) 1 L of distilled water) was mixed with plant-derived natural substances to inoculate the pathogenic bacteria to confirm the growth of the pathogenic bacteria. The origin of more than 20 species of plants After a review of the antibacterial activity of natural substances, gallnut (Galla chinensis ) was selected as a basic substance of the present invention by exhibiting antibacterial activity through the result that the bacterium of Ashidoborax Abene Hospital could not grow at a concentration of 1,000 ppm (Figs. 1 to 3).

Example  Two: Echinacea  Antibacterial activity according to the input ratio

Galla chinensis ) and Echinacea purpurea were tested for antimicrobial activity against A. avenae subsp. citrulli hospital bacteria at various mixing ratios (Table 1). As a result, it was confirmed that the addition of dwarf alone to the mixture of dwarf and echinacea was superior to the antibacterial activity.

Antibacterial activity according to the input ratio of dwarf and echinacea number Raw input ratio (%) Acidoborax Abeta species Citrulli Antibacterial activity 2000 ppm 1000 ppm 500 ppm One 100 +++ ++ + 2 90 + + + 3 80 ++ + + 4 70 ++ + + 5 60 + + - 6 50 + - - 7 40 + - - 8 30 - - - 9 20 - - - 10 10 - - - 11 0 - - -

+: Ashidobolox Abeta species Citrus growth inhibition 1mm or less

++: Ashidobolox Abeta species Citrus growth inhibition 1 ~ 3mm

+++: Ashidobolax Abeta species Citrus growth inhibition 3mm or more

-: Ashidobolax Avetacea Species No growth inhibition of citrus fruit

Example  3: Antibacterial activity of the formulation mainly composed of obtusa

( G. chinensis ), which exhibits strong antibacterial activity against fruit rot disease, is mixed with various auxiliary materials such as surfactants, extenders and emulsifiers which are low in the effect of chemical reaction, Respectively. The crude extract was prepared by pulverizing 10 kg of dwarf and then extracting twice with 100% methanol. The extract was concentrated under reduced pressure at 40 ℃.

A. avenae subsp. Citrulli Hospital sterilized or inhibited bacteria contaminated with soil disinfection, contamination of soil disinfection, thermal insulation, and tubing materials in potting facilities of seedling plants and seedling plants, and cultivation of crops Liquid formulations for controlling juice transmission were formulated with additives to prevent coagulation and freezing (Table 2).

Composition of Liquid Formulation Based on Rhizoma Extract Formulation
number Obovate extract
Input ratio Glycerol ethanol PEG-400 CO-40 GMO-350 NP-10 water BFB-1 30 30 - - - - - 40 BFB-2 30 - - - - - - 70 BFB-3 35 5 40 - - - - 20 BFB-4 35 5 30 - - - - 30 BFB-5 35 - - 10 - - - 55 BFB-6 35 - - - 10 - - 55 BFB-7 35 - - - - 10 - 44 BFB-8 35 5 - 10 - - - 50 BFB-9 35 - 40 - - - 5 20 BFB-10 35 - 40 - - - - 25

To confirm the effect of the antimicrobial activity on the liquid formulation of A. avenae subsp. Citrulli , the test agent diluted with 500 ppm, 1,000 ppm and 2,000 ppm of each formulation in a medium was applied to a disk paper (Φ8 ㎜), 150 μl of each was inoculated. As a result, a liquid formulation BFB-3 having excellent antibacterial activity was selected (Table 3).

Antibacterial activity assay of formulations based on rhizome extract Formulation Number Acidoborax Abenet antibacterial activity 2,000 ppm 1,000 ppm 500 ppm BFB-1 ++ + + BFB-2 + + + BFB-3 +++ ++ + BFB-4 +++ ++ + BFB-5 ++ + + BFB-6 ++ + + BFB-7 ++ + + BFB-8 +++ + + BFB-9 ++ ++ + BFB-10 ++ + +

+: Ashidobolox Abeta species Citrus growth inhibition 1mm or less

++: Ashidobolox Abeta species Citrus growth inhibition 1 ~ 3mm

+++: Ashidobolax Abeta species Citrus growth inhibition 3mm or more

-: Ashidobolax Avetacea Species No growth inhibition of citrus fruit

Example  4: Physical stability of formulations based on obovate

The physical stability of liquid formulation BFB-3, which exhibited strong antimicrobial activity against asidobolax avenae, was investigated. The pH, specific gravity, and stability of the liquid formulation BFB-3 (FIG. 4) having a high antibacterial activity were examined. 1 g of the liquid formulation was added to a beaker containing 100 ml of distilled water, and the pH was measured using a pH meter. The specific gravity was measured by filling the liquid formulation into a 100 ml measuring cylinder and measuring the specific gravity. Water solubility dropped from 10 cm from the surface of the water when there was no flow of liquid form BFB-3 in the beaker containing 100 ml of distilled water. At this time, wetting time was measured from the dropping time to confirm the hydration ability. In addition, the stability of the liquid formulation was investigated by observing changes in the material by standing in an oven at 50 캜 for 4 weeks.

As a result, the physical stability of the liquid formulation BFB-3, which is the major component of the extract, was confirmed. The physical stability was confirmed, and it was found stable at 50 ° C without any substance change (Table 4).

Physical Stability of Liquid Formulation BFB-3 with Larval Extract as the Main Component Survey item Liquid formulation BFB-3 pH 2.97 importance 1.08 Hydration ability 5 seconds stability No material change

Example  5: In the greenhouse In fruit rot  Anti-bacterial activity test

Ashido Lovers Abene In order to investigate the antibacterial activity of liquid formulations based on the extracts of Rhus verniciflua and Rhus verniciflua, which have strong anti - bacterial activity against Rhizoctonia spp.

(1) A. avenae subsp. Citrulli Establishment of disease condition of bacterial fruit rot in watermelon of hospital bacterium

A. avenae subsp. Citrulli In the greenhouse of the hospital bacterium Bacterial fruit rot disease To determine the onset condition, watermelon (seedling) seeds were dipped in hospital bacterium and seeded, The bacterial culture was sprayed with the leaves to maintain the humidity of 70% at 30 ° C, and the occurrence of the diseases was compared. As a result of examining the disease occurrence conditions, the symptoms of both seed immersion and foliar spraying were confirmed. However, in the seed dipping method, the watermelon seedlings were found to appear after 45 days at 30 ° C. When the seedlings were treated with foliage, A symptom was confirmed after 30 days (Fig. 5).

(2) Effect of controlling watermelon bacterial fruit rot in greenhouse

The pot test was carried out on the seedlings of pine and cucumber watermelon (honeysuckle). The extract of G. chinensis , a plant-derived natural substance with high antibacterial activity in watermelon seedlings, 5,000 ppm, 2,000 ppm and 1,000 ppm, foliar treatment of 10 ml per week, and dilution of the liquid formulation BFB-3 to 2,000 ppm and 1,000 ppm, respectively. The bacterium was inoculated with 10 ml of the culture broth of A. avenae subsp. Citrulli hospital (10 ⁸ cfu / ml) with 10 ml of leaf spray per day. Then, the bacteria were cultured at 30 ° C for 30 days to observe bacterial fruit rot disease symptoms, and the antibacterial activity against watermelon seedlings was examined. In the control, the distilled water was sprayed with distilled water instead of dwarf, and the antibacterial activity was compared by treatment of golden bell product, which is a chemical synthetic pesticide, Kasugamycin as a positive control.

The antibacterial activity against the bacterial fruit rot rot fungi was examined at 30 days after the treatment with the extract of G. chinensis in the seedlings of Pak and Cucumber watermelon. (Table 5). In addition, antibacterial activity against bacterial fruit rot rot fungi was examined 30 days after the liquid formulation BFB-3 was treated in the seedlings of Pak and crops watermelon. As a result, 69.2% of the antibacterial activity was observed at 2,000 ppm of the liquid formulation 6).

Of G. chinensis extract against bacterial fruit rot disease Antibacterial effect Test agent Treatment concentration Number of sickling seeds
(15 per week) Antibacterial activity (%) Obovate extract 5,000 ppm 2 83.3 2,000 ppm 3 75 1,000 ppm 3 75 Kasugamycin
(Kasugamycin) 250 ppm 10 25 Control (distilled water treatment) - 12 -

Antibacterial effect of liquid formulation BFB-3 on bacterial fruit rot disease Test agent Treatment concentration Number of sickling seeds
(15 per week) Antibacterial activity (%) Liquid formulation BFB-3 2,000 ppm 4 69.2 1,000 ppm 6 53 Kasugamycin
(Kasugamycin) 250 ppm 10 23.07 Control (distilled water treatment) - 13 -

Example  6: Analysis of active ingredient substances derived from obtusa extract

The antimicrobial active substance was isolated from the rhizome extract and the structure of the compound was determined. The crude extract was prepared by pulverizing 10 kg of dwarf and then extracting twice with 100% methanol. The extract was concentrated under reduced pressure at 40 ℃. The obtained methanol extract of dicotyls was suspended in 1 L of distilled water and subjected to solvent fractionation successively using n-hexane, chloroform (CHCl ₃ ), ethyl acetate (EtOAc) and n-butanol (BuOH). A. avenae subsp. Citrulli The substances that inhibit the growth of the bacteria were selected and analyzed by HPLC. The results were compared with those of the methyl gallate standard This antimicrobial active substance was identified as methyl gallate.

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Claims (10)

Bacterial fruit blotch containing a mixture of methanol extract of Galla chinensis , glycerol, ethanol and water in an amount of 30 to 40: 4 to 6:30 to 40: 20 to 30 volume ratio as an active ingredient, (2). delete delete delete Claim for according to item 1, wherein the watermelon bacterial fruit rot (Bacterial fruit blotch) organisms are know walk flux Oh Venetian subspecies sheet rulri (Acidovorax avenae subsp. Citrulli) of watermelon bacterial fruit rot (Bacterial fruit blotch), characterized in that the control Composition. delete delete delete delete A method for controlling a bacterial fruit blotch, comprising the step of treating a bacterial fruit blotch controlling composition of claim 1 or 5 to a foil, a crop or a seed.

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