KR101892470B1 - Natural Disinfectant - Google Patents

Abstract

Disclosed is a natural disinfectant comprising a natural microorganism as an effective ingredient and an effect enhancing additive capable of enhancing the effect of the microorganism, thereby maximizing sterilizing power by the microorganism. The present invention relates to a method for producing Streptomyces sp . ) Or a culture medium of Bacillus sp. As an active ingredient and has an active activity against cucumber powdery mildew or strawberry gray mold disease.

Description

Natural Disinfectant

The present invention relates to a natural disinfectant, and more particularly, to a natural disinfectant containing natural microorganisms as an effective ingredient and including an effect enhancing additive capable of improving the effect of the microorganism, will be.

Recently, environmental problems are directly related to the survival of human beings, especially the food problem, and there is a growing interest in the world.

Domestic pesticide use amounted to 1,205kg / km2 in the mid-1990s, second only to Japan among OECD member countries, 4.6 times that of OECD average usage amount of 263kg, 48 times that of New Zealand (25kg) In order to solve these problems, the '92 Rio Environmental Conference has decided to reduce the use of organic synthetic pesticides by 50% by 2004, In 1997, in the United States, 46% of the use of organic synthetic insecticides, in particular 68% in vegetable and fruit growing, was converted to alternativepesticides. In the United States, 14% of fruit and vegetable fields and 6 % Are conducting pest control ('98, USDA-NASS) using insect pathogenic fungi and nematode insects. This biological control is environmentally friendly Since the pesticide concentration can be continuously suppressed to the agricultural ecosystem as a means of control, many studies are being carried out at a national level in foreign countries, and the biological development of the pest is completed by a single active ingredient. The proportion of the total sales is expected to increase gradually.

In the past two decades, it has become possible to reduce environmental risks due to the development of biological control technology in the world. However, since the abuse of chemical pesticides in Korea has seriously damaged agricultural crops, (Agricultural Research and Technology Institute, 2006). In order to improve this, it is necessary to comply with the safe use standards of pesticides at individual farms It is important to develop and distribute products such as eco-friendly crop protection products and technologies using them.

In addition, as a free trade agreement (FTA) between Chile, Korea, and an EU FTA has been concluded, it is clear that production is necessary to effectively cope with such low-cost imported agricultural products. In order to produce such eco-friendly agricultural products continuously, it is necessary to improve income of environment-friendly farmers' growers and secure stable profitability. To this end, technologies for producing environmentally friendly agricultural products should be developed and widely distributed.

In terms of economic and industrial aspects, the development cost of environmentally friendly microbial pesticides may be relatively more economical than that of chemical pesticides, because toxicity and residue related tests, which make up the bulk of chemical pesticide development costs, are relatively palliative, It is believed that replacing existing chemical pesticides, which depend on imports, will greatly contribute to the recovery of food sovereignty as well as the import substitution effect.

However, there are many problems to be solved in the development of microbial pesticides because the pesticides have no merits and they have disadvantages. Especially, there are problems such as a short circulation period and a long period of time, a narrow application range, a problem affecting only a specific object, a delay in the occurrence of the effect, and low efficacy.

Korean Patent No. 13255599 discloses an environmentally friendly composition for controlling cucumber powdery mildew and cucumber fungus, and a method for producing the same. According to the present invention, an eco-friendly antiseptic agent produced by mixing rosin oil and eucalyptus oil exhibits a sterilizing effect, but has a problem that the sterilizing power is weak due to the nature of the oil and the duration is short.

Korean Patent No. 0862807 discloses a method for producing an agricultural fungicide for agricultural use using propolis. In the present invention, the propolis powder is used for the agricultural sterilizing agent, but the sterilizing power is weak and the duration is short.

In addition, when the environment-friendly crop protection agent and fertilizer are developed, it can be applied to the commercialization of many useful microorganisms that can be used in the past, and it can be a basis for securing various microorganism pesticide raw materials and products which are competitive in the domestic market. There is an advantage that the product can be distributed.

The development of environmentally friendly crop protection and effective production processes is a key area in the next generation biotechnology industry. The development of a 21st century crop protection system is a very important task in Korea at the present time, which should secure international competitiveness and become a developed country of science and technology. And to be able to integrate into the export market.

From the technical point of view, the technology of formulation of environmentally friendly microbial pesticide / fertilizer is not beyond the basic level in the domestic as well as the world. In particular, in formulation technology, selection and manufacturing process of additives, microorganisms and crops with insecticidal / It is very difficult for microorganisms to maintain their effectiveness and stability until stable microorganisms can be stably sprayed on the field, so that if the technology of this research is developed, it will be recognized as a world-class technology. The effect of the effect on the actual environment is weak.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a natural disinfectant which can have a higher effect than existing natural disinfectants, including natural microorganisms having sterilizing power and additives for enhancing effects of the microorganisms I want to.

In order to solve the above-mentioned problems, the present invention relates to a method for producing Streptomyces sp . ) Or a culture medium of Bacillus sp . As an active ingredient and has an active activity against cucumber powdery mildew or strawberry gray mold disease.

Oregano oil, polyalkylene oxide modified heptamethyltrisiloxane (PMH2), and castor oil; as additives for enhancing the effect; Characterized in that the formulation point further comprises a zero surfactant.

The natural disinfectant according to claim 1, wherein the natural disinfectant comprises 30-50% of actinomycetes or bacterium culture liquid, 10-30% of oregano oil, 1-10% of PMH2, 10-30% of castor oil and 1-10% of surfactant Lt; / RTI >

And further comprising kaolin gum as a thickening agent, a surfactant as a dispersing agent, and kaolin as a fixing agent.

Also, the natural bactericides include a culture broth of actinomycetes or bacillus bacteria containing 60 to 90% of xanthan gum at 0.01 to 1%, surfactant at 1 to 10%, and kaolin at 10 to 30%.

According to the present invention, since the natural microorganism having sterilizing power and the effect enhancing additive for further enhancing the effect of the microorganism are contained, the duration of the sterilizing effect is long, and the effect is higher than that of the existing natural microbicide, It is useful for growing agricultural products.

1, JA (II) -10, B, JR-45, C, KJA-35,
FIG. 2 is a graph showing the results of the minimum inhibitory concentration test for strawberry gray mold,
FIG. 3 shows the results of the minimum inhibitory concentration test for JA (II) -10, wherein A, B, and C were 103 cells / ml, 104 cells /
FIG. 4 is a photograph of the antifungal activity test of antibiotic-resistant strain JA (II) -10 in an antagonistic microorganism of Staphylococcus aureus.
FIG. 5 is a photograph showing the strawberry plant decay of JA (II) -10 strain,
FIG. 6 is a photograph showing the appearance of the strawberry gray mold pathogen in the treatment distinction,
FIG. 7 shows the phylogenetic position of the strain JA (II) -10, which is an antifungal activity-resistant strain against Staphylococcus aureus
FIG. 8 is a schematic diagram showing a method for selecting a strain of antifungal activity against cucumber powdery mildew fungus,
Fig. 9 is a photograph showing a strain of antibiotically active cucumber powdery mildew in cucumber by an in-
FIG. 10 is a graph showing the results of the in vitro fertilization sp . Comparison of antimicrobial activity of GG95 on cucumber powdery mildew
FIG. 11 is a graph showing the effect of controlling antibiotic activity of a cucumber powdery mildew by antimicrobial activity,
FIG. 12 is a schematic diagram of an experimental method for evaluating the antimicrobial activity of a cucumber powdery mildew fungus,
FIG. 13 is a photograph showing the result of the antibacterial activity microorganism test for each treatment,
Fig. 14 is a graph showing the incidence of cucumber powdery mildew of yeast bacteria by antimicrobial activity treatment (packing test)
Fig. 15 is a graph showing the results of measurement by Bacillus sp . GG95, Cucumber Powdery Mildew, Antimicrobial Activity,
Fig. 16 is a graph showing the degree of onset of cucumber powdery mildew disease (antimicrobial activity)
FIG. 17 is a graph sp . Antimicrobial activity test of cucumber powdery mildew fungus of GG95 strain.
18 shows the phylogenetic location of the strain JH-19, which has excellent antimicrobial activity against cucumber powdery mildew,
Fig. 19 shows the phylogenetic location of the strain GG95, which is an antimicrobial active strain against cucumber powdery mildew,
Fig. 20 shows the results of the antimicrobial activity test (environment horn method) of the environmentally-friendly extract against the field strain,
21 shows the results of the antibacterial activity test for the gray mold fungus Botrytis cinerea according to the concentration of Oregano oil
22 is a graph showing the results of the incidence of cucumber powdery mildew by microbial treatment concentration,
FIG. 23 is a graph showing the effect of microbial and natural oil mixture treatment on cucumber powdery mildew,
24 is a graph showing the effect of microbial mixture treatment on cucumber powdery mildew in a facility house
25 is a graph showing the effect of the antibacterial microorganism treatment on strawberry gray mold disease.

Hereinafter, preferred embodiments of the present invention will be described in detail. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. Throughout the specification, when an element is referred to as " including " an element, it means that it can include other elements, not excluding other elements, unless specifically stated otherwise.

The inventors have in natural fungicide, was a conventional fungicide effect of the short duration simple, and also noted that the sterilizing power is not high, and results of the working example In order to solve this problem, Streptomyces (Streptomyces sp . ) Or Bacillus sp . ) Culture broth as an active ingredient, it is possible to solve such a problem, leading to the present invention.

Therefore, the present invention relates to a method for producing Streptomyces sp . ) Or Bacillus sp . ) Culture medium as an active ingredient and having an active activity against cucumber powdery mildew or strawberry gray mold disease.

The actinomycetes or bacillus strains which are effective for cucumber powdery mildew or strawberry gray mold disease used in the present invention can be used without limitation, but preferably Streptomyces acidiscabies (JA (II) -10), Streptomyces sp . JR-45, Streptomyces sp. KJA-35, Streptomyces sp . KJA-2, Streptomyces sp . KJA-30, Streptomyces sp . JH-19 or Bacillus sp . GG95. More preferably Streptomyces acidiscabies (JA (II) -10), or Bacillus sp . GG95.

Also, in the present invention, Oregano oil, polyalkylene oxide modified heptamethyltrisiloxane (PMH2) and castor oil are used as an effect enhancing additive; And may further be characterized in that the formulation point further comprises a zero surfactant. Preferably, the additive is added in an amount of 30 to 50% of actinomycetes or bacterium culture liquid, 10 to 30% of oregano oil, 1 to 10% of PMH2, 10 to 30% of castor oil, 1 to 10 of surfactant %. ≪ / RTI > More preferably, it may contain 40 to 50% of actinomycetes or bacterium culture liquid, 23 to 27% of oregano oil, 3 to 7% of PMH2, 15 to 25% of castor oil and 3 to 7% of surfactant.

Further, the present invention may further comprise kaolin gum as a thickening agent, a surfactant as a dispersing agent, and kaolin as a fixing agent. The additive may be added to make a liquid agent, and it may preferably contain 0.01 to 1% of 60% to 90% of xanthan gum of a actinomycetes or a bacterium culture liquid, 1 to 10% of surfactant, and 10 to 30% of kaolin. More preferably, the culture broth of actinomycetes or Bacillus bacteria may contain 70 to 80% of xanthan gum 0.05 to 0.2%, surfactant 3 to 7%, and kaolin 15 to 25%.

In addition, the present invention may further comprise guano as an effective adjuvant, and molasses, a corrosive acid and a surfactant as a molding aid. Preferably, the additive is added in an amount of 5 to 15% of actinomycetes or bacillus culture media, 80 to 95% of granular guano, 0.1 to 2% of molasses, 1 to 5% of corrosive acid, 0.01 to 0.05% of surfactant, . More preferably, it may contain 7 to 10% of actinomycetes or bacterium culture liquid, 85 to 92% of granular guano, 0.8 to 1.5% of molasses, 1 to 3% of corrosive acid, and 0.01 to 0.05% of surfactant.

In addition, the present invention may further comprise a guano powder as an effect supplement, xanthan gum as a thickening agent, a surfactant as a dispersing agent, and citric acid as a stabilizer. Preferably, the additive is added to make a liquid wettable powder. Preferably, the additive is added in an amount of 50 to 70% of actinomycetes or bacterium culture liquid, 10 to 50% of guano powder, 0.01 to 0.5% of xanthan gum, 1 to 10% of surfactant, 5%. ≪ / RTI > More preferably, it may contain 60 to 70% of actinomycetes or bacterium culture liquid, 25 to 35% of guano powder, 0.05 to 0.2% of xanthan gum, 3 to 7% of surfactant, and 1 to 3% of citric acid.

Hereinafter, a specific embodiment of the present invention will be described.

○ Strawberry gray fungus pathogen ( Botrytis cinerea ) On the antimicrobial activity Outstanding strain  Selection

A total of 300 strains were collected from 50 bamboo forest litter samples, 50 strains from rhizosphere soil, 50 strains from livestock manure compost and 150 strains from heavy metal contaminated soil. The antibacterial activity of 300 strains of actinomycetes was investigated against strawberry gray mold pathogen ( Bortrytis cinerea ). Of the 50 strains isolated from the litter samples, 30 strains showed antimicrobial activity against strawberry gray mold pathogens. A total of 145 strains showed antimicrobial activity against 44 strains in rhizosphere soil, 36 strains in compost, and 35 strains in soil contaminated with heavy metals (Table 1). Three strains of actinomycetes (JA (II) -10, JR-45, and KJA-35), which have excellent antimicrobial activity against Staphylococcus aureus fungi, were selected as excellent strains among the actinomycetes showing these antibacterial activities (FIG. 1).

Sample Total isolates Antibacterial isolates Bamboo litter 50 30 Bamboo rhizosphere soil 50 44 Compost 50 36 Chrome polluted soil 100 20 Mercury polluted soil 50 15 Total 300 145

○ Strawberry gray mold pathogen  Minimum inhibition concentration test ( Minimum Inhibitory  Concentration, MIC )

Strawberry gray mold pathogens ( Botrytis In order to investigate the minimum inhibitory concentrations of JA (II) -10, JR-45 and KJA-35 strains, which showed excellent antimicrobial activity against T. cinerea , the concentrations of strains were 103, 104, 105 and 106 cells / broth, TSB), and seeded on an 8 mm paper disc. The growth inhibition effect was examined by culturing the strain in an optimum condition of a strawberry gray mold pathogen. In the case of JA (II) -10 strain, which had the highest antimicrobial activity, the inhibitory effect was more than 12 mm at a concentration of 103 ~ 106 cells / ㎖. KJA-35 strain was inhibited at a concentration of 105 ~ 106 cells / And the JR-45 strain showed an inhibitory effect of 3 mm at a concentration of 106 cells / ml (Fig. 2). As a result, a JA (II) -10 strain excellent in antibacterial activity even at a concentration of 103 cells / ml was finally selected (FIG. 3).

○ Strawberry gray fungus pathogen  Antimicrobial activity Outstanding strain JA (II) -10 Strain (In-Flight Test)

In order to induce the natural development of the staphylococcus aureus fungi, strawberry was placed in a transparent case prepared by repeating 3 repeats 10 times without washing process. In the treatment of JA (II) -10 strain, it was sprayed at a concentration of 106 CFU / ㎖ and cultured for one week in a culture room adjusted to 23 ℃. The effect was evaluated by counting the infected fruits infected with the gray mold pathogens.

Strawberry fruit fungus infected with Staphylococcus aureus fungus was observed in each treatment on the 2nd day of the inoculation. The infection rate of each treatments on the 4th day of inoculation was 63% in the untreated area, whereas in the JA (II) And the rate of disease progression was decreased. As a result, JA (II) -10 strain showed a decrease in onset of strawberry gray mold pathogens and delayed the onset of the disease (FIG. 4).

○ Strawberry gray fungus pathogen  Antimicrobial activity Outstanding strain JA (II) -10 < / RTI > Port test )

Strawberry plants were inoculated with sterilized water and JA (II) -10 strains for each treatments, and the degree of weakness was evaluated by examining symptoms on the leaves. The results of counting abnormalities such as spots, atrophy, sulphurisation, and ogal appearing on the leaf for 2 days after the inoculation were evaluated for the degree of the weakness of each leaf in the untreated and JA (II) -10 strain treatments, (II) -10 strains were selected as the antimicrobial active strains of the strawberry gray mold pathogens (FIG. 5).

○ Strawberry gray fungus pathogen  Antimicrobial activity Outstanding strain JA (II) -10 Strain (Packing test)

For the application test of JA (II) -10 strain, which showed the most effective antimicrobial activity against strawberry gray mold in the in vitro drug test, strawberry gray mold pathogens were inoculated to the growing strawberry plant in the test specimen of the strawberry test center of Chungnam Agricultural Research Institute. .

The treatments were divided into treatments (sterilized water), JA (II) -10, and control (ecojet) treatments, and 30 juveniles of strawberry plants per treatment were examined to examine the effect of controlling the infection of strawberry gray mold pathogens Respectively. As a result, the inhibition rate of gray mold fungi to strawberry on JA (II) -10 strain was 19.9% in control, whereas it was 8.3% in JA (II) -10 strain. (Table 2). In the case of the control, it was 58.3 as compared to 33.3 of the control. As a result, it was found that the AUDPC was significantly reduced to 144 compared to 391 for the control and 234 for the control. JA (II ) -10 strain was found to be superior to the strawberry fungus pathogen-inhibiting effect (Fig. 6).

Treatment Disease rate (%) The control AUDPC ² Control 19.9a ¹ - 391 JA (II) -10 8.3b 58.3 144 Contrast agent 13.3ab 33.3 234

○ Strawberry gray mold pathogen  Antimicrobial activity against Outstanding strain JA (II) -10 strain classification

The 16S rRNA gene sequence (1,422 bp) of the strain JA (II) -10, which is an antimicrobial activity against strawberry gray mold, was determined and EzbioCloud (http://www.ezbiocloud.net/eztaxon) Location. Each nucleotide sequence alignment using the CLUSTAL W algorithm, and represented a MEGA 5.0 results, Streptomyces acidiscabies ATCC 49003T and 99.7% homology to create the phylogenetic tree using a program based on the neighboring coupling method Streptomyces acidiscabies (Fig. 7).

○ Cucumber Powdery mildew  Selection of Antimicrobial Active Microorganisms ( Seedling assay )

A total of 25 strains of the IMER microorganism ecosystem research center of Mokwon University were selected as the antifungal active strains of the powdery mildew fungus according to the seedling growth method of cucumber powdery mildew (Table 3). The test strain was cultured in TSB liquid medium at 25 ° C with shaking at 150 rpm for 3 days, adjusted to 106 CFU / ml, and the infection index was examined after 7 days in a thermostat at 25 ° C (FIG. 8) . Streptomyces showing an onset index of 40% or less sp . KJA-2, Streptomyces sp . KJA-30, Streptomyces sp . JH-19 and Bacillus sp . GG95 strain was selected as a strain of antimicrobial activity against cucumber powdery mildew (Fig. 9, Fig. 10)

NO. IMER carrying strains Strain name Cucumber powdery mildew
Incidence (%) One Streptomyces tsukionensis JA (II) -45 86.7 2 Streptomyces sp. JA (II) -50 88.3 3 Saccharopolyspora sp. MF142-21 91.2 4 Streptomyces sp. KJA (Y) -5 90.1 5 Amycolatopsis sp. MF156-22 84.5 6 Streptomyces sp. MM5-23 86.4 7 Streptomyces flavotricini MF65-13a 88.9 8 Kribbella karoonensis MF142-3 93.2 9 Strptomyces ciscaucasicus MF142-8 94.2 10 Streptomyces sp. MF67-6 81.6 11 Streptomyces flavotricini MF65-13b 88.9 12 Streptomyces sp. KJA-2 39.4 13 Streptomyces sp. KJA-30 38.6 14 Streptomyces sp. MF31-4 87.6 15 Lentzea kentuckyensis MF156-40 80.8 16 Streptomyces sp. KJA-35 92.4 17 Streptomyces sp. KJA-26 90.6 18 Streptomyces acidiscabies JA (II) -10 90.8 19 Streptomyces sp. JH-4 83.7 20 Streptomyces sp. JH-11 81.0 21 Streptomyces sp. JH-19 29.8 22 Streptomyces olivochromogenes JH (II) -9 78.6 23 Streptomyces olivochromogenes JR-36 83.3 24 Streptomyces sp. JR-39 76.9 25 Bacillus sp. GG95 20.1

○ Cucumber powdery mildew Antifungal activity microorganism test (pot test method)

Selected Streptomyces sp . KJA-2, KJA-30, JH-19 and Bacillus sp . The antimicrobial activity of the cucumber seedlings with cucumber powdery mildew was investigated in 3, 7, and 10 days after GG95.

Antimicrobial activity strain KJA-2 showed 39.8% morbidity after 10 days and the control value was 54.2%. The strain KJA-30 showed 30.9% of disease and the control showed 64.4%. The strain JH-19 showed 25.8% after 10 days and the strain GG95 showed 20.1% after 10 days. 80.9%, showing the most excellent control effect (FIG. 11).

○ Cucumber powdery mildew Antimicrobial active microorganism test (Outer packing test method)

To evaluate the antimicrobial activity of cucumber powdery mildew, antimicrobial activity of cucumber powdery mildew was tested in 10 treatments of 5 treatments in a cucumber house farmhouse in Iin-myeon, Ginju, Gyeonggi-do, where the cucumber powdery mildew was developed. The test strain was cultured in TSB liquid medium for 3 days at 25 rpm at 150 rpm After culturing the microbial broth at 106 CFU / ml, the cucumber powdery mildew of cucumber powdery mildew was sprayed three times at intervals of 7 days, and the onset index of the mildew powdery mildew was divided into the control, the culture medium and the antimicrobial active microbial treatment (Fig. 12).

In the control group, the disease progression rate of cucumber powdery mildew disease bacterium was 90.1%, whereas in the test group treated with the antimicrobial activity strain, the progress of the disease was delayed to 21.9 to 53.9% (FIG. 13). In particular, the JH-19 strain showed a control value of 66.8%, indicating that it was the most effective antimicrobial active strain for Cucumber powdery mildew (Fig. 14).

Cucumber powdery mildew Bacterium Bacillus sp . For the evaluation of the antimicrobial activity of the cucumber powdery mildew of the cucumber powdery mildew (GG95) strain, the cucumber powdery mildew cucumber was tested for 10 treatments in 5 treatments at a cucumber house farmhouse in Geumnam-myeon, Sejong City (Fig. 15).

The test strain was cultured in TSB liquid medium at 25 ° C for 3 days with shaking at 150 rpm. After adjusting the microbial culture to 106 CFU / ml, the cucumber powdery mildew of cucumber powdery mildew was sprayed 4 times at intervals of 7 days, The onset index was evaluated by dividing into the non-treatment area, the culture medium and the microbial treatment area with antibacterial activity.

On the other hand, the incidence of cucumber powdery mildew was 96.2%, while the antimicrobial activity strain Bacillus sp . In the treated group treated with GG95, 74.6% of the control group was selected, and the strain was finally selected as a strain excellent in antibacterial activity against cucumber powdery mildew (FIG. 16).

Selected Streptomyces sp . KJA-2, KJA-30, and JH-19 strains were treated with antimicrobial activity against the cucumber leaf which had developed cucumber powdery mildew, and 21.9 ~ 53.9% of them were found.

Cucumber Powdery Mildew Bacillus sp . As a result of the evaluation of the efficacy of cucumber powdery mildew fungus on GG95, it was selected as the most effective antimicrobial active strain for Cucumber powdery mildew disease (Table 4, Fig. 17).

The control Non-treatment Treatment KJA-2 KJA-30 JH-19 GG95 - 40.1ab ¹ 47.5ab 66.8a 27.9b

¹ Treatment means followed by different letters differ significantly (Fisher's protected LSD, P = 0.05)

○ Cucumber Powdery mildew  Systematic analysis of antimicrobial active microorganisms

The 16S rRNA gene sequence of JH-195 was determined and the homology of NCBI / RDP / GenBank was detected. JH-19 showed 100% homology with Streptomyces xanthocidicus NBRC 13469 ^T (AB184427) and was identified as Streptomyces xanthocidicus (Fig. 18).

When the 16S rRNA gene sequence of GG95 was determined and the homology of NCBI / RDP / GenBank was determined, Bacillus subtilis subsp. was identified as Bacillus subtilis with 99.9% homology to spizizenii NRRL B-23049 ^T (CP002905) (Fig. 19).

○ Formulation  Development of technology

In the case of microbiological probiotics, the stability is drastically lowered under general cultivation conditions and there is a marked difference in effect depending on various environmental conditions. Therefore, the addition of stabilizers and adjuvants through the formulation process has been aimed to enhance the effect to the extent that it can be commercialized.

A) Selection of auxiliaries for efficacy improvement

The purpose of this study was to investigate the effect of natural materials which can be used as organic agricultural material for the control of cucumber powdery mildew and strawberry gray mold in order to secure the stability of the microorganism preparation in the field and to improve the rapid effect.

(1) Selection of eco-friendly extracts for controlling powdery mildew

- Test sample: 80% Eucalyptus oil, 65% Clove oil, 60% Oregano oil, clove extract (0.3% Eugenol)

- Target crops: Cucumber (whitefish)

- Treatment content: Soak seedlings so that the whole seedlings can flow with 1000 times diluted solution

- Investigations: 7 days after the treatment,

- Results: As shown in Table 5, most of the eco-friendly extracts showed a high control effect of over 80%. Especially, Oregano oil and Clove oil showed higher control effect than other commercialized extracts, and were used as samples for determination of the first selection and optimum concentration.

Treatment Treatment concentration Cucumber powdery mildew prevention effect DMRT Outbreak rate (%) Control (%) No treatment - 55.0 + - 5.0 - e 80% Eucalyptus oil 1000 times 8.8 ± 1.3 84.1 CD 65% Clove oil 1000 times 6.3 ± 1.3 88.6 ab 60% Oregano oil 1000 times 5.0 ± 1.5 90.9 ab 0.3% Eugenol 1000 times 10.0 ± 2.5 81.1 CD

(2) Determination of effective concentration of eco-friendly extract against powdery mildew

The first selected plant extracts against cucumber powdery mildew were further tested for cucumber powdery mildew in each diluted solution to determine the economical effective concentration.

- Test sample: 60% Clove oil, 60% Oregano oil, 60% sulfur WP (control agent)

- Target crops: Cucumber whitefish

- Treatment content: dilution of each treatment lot is diluted 1000 times, 2000 times, 4000 times,

- Investigations: 7 days after the treatment,

- Results: As shown in Table 6, the control effect of Oregano oil against powdery mildew was higher than that of Clove oil. In case of 60% Oregano oil, the control effect was better than 2000 times. Therefore, the formulation test was conducted based on the effective concentration of 300 ppm or higher.

Treatment Treatment concentration Cucumber powdery mildew prevention effect Outbreak rate (%) Control (%) No treatment - 52.9 ± 9.6 - 65% Clove oil 1000 times 13.3 ± 2.7 74.9 2000 times 15.2 ± 3.3 71.3 4,000 times 24.1 ± 5.0 54.4 60% Oregano oil 1000 times 9.8 ± 1.9 84.3 2000 times 10.6 ± 2.8 80.0 4,000 times 15.1 ± 6.2 71.5 * 60% sulfur (contrast agent) 2000 times 5.8 ± 1.0 89.0

* The effect was verified by using [Huang Star] as the reference drug.

(3) Selection of eco-friendly extracts for gray mold disease

The extracts with indoor antimicrobial activity against the gray mold were firstly selected and used as a supplement for the microbial effect.

- Test sample: 80% Eucalyptus oil, 65% Clove oil, 60% Oregano oil, clove extract (0.3% Eugenol)

- Test method: Evaluation of antibacterial activity against representative horticultural diseases such as Botrytis cinerea

- Treatment content: After sterilization of PDA (+; antibiotic added) culture medium, each sample is diluted 100 times, and inoculated into each culture medium with Botrytis sp.

- Investigations: After 5 days of incubation at 28 ° C, mycelial growth of mycorrhizal fungi was investigated after inoculation with the field cultures.

Results: As shown in FIG. 20, the antimicrobial activity of 60% Oregano oil 100-fold dilution was the best, and ORE>CLO>EU> EUC showed the best effect.

(4) Determination of Effective Concentration of Environmentally-friendly Extracts for Gray Mold Fungi

To evaluate the efficacy of Oregano oil, the most effective antimicrobial activity against the gray mold, we evaluated the antimicrobial activity by concentration.

- Test sample: 60% Oregano oil

- Test method: Evaluation of antimicrobial activity against Botrytis cinerea and virulent strain

- Treatment content: PDA (+; antibiotic added) 60% Oregano oil was diluted 500 ~ 4000 times in sterilized medium to prepare antimicrobial medium

- Investigation: Culture for 5 days after the inoculation of the pathogenic bacteria

Results: As shown in FIG. 21, the antimicrobial activity of 60% Oregano oil was shown up to 4000 times and the antimicrobial activity was exhibited even at a low concentration gradient of 1000 to 4000 times. In addition, it does not show a large difference according to the concentration from 1000 times dilution to 4000 times dilution. Therefore, it can be applied even at a low concentration of 4000 times or more. Therefore, formulation of 150 ppm or more was ensured at an effective concentration.

B) Oil-in-water suspensions,

(1) Emulsifier selection

Oregano oil, which has the best effect on cucumber powdery mildew and gray mold disease, should be prepared so that raw materials can be used with water and diluted with a certain emulsifier. Therefore, the process of selecting surfactants optimized for Oregano oil was studied in advance.

- Emulsifier selection of Oregano oil

: EPA List used for agriculture 4A, 4B grade safe emulsifier is applied within 10%, 20%

- Selected emulsifiers are listed in Table 7 below.

division CAS No. EPA List Degree of emulsification Remarks 10% 20% AG-PLG 4B 4B ○ ◎ Emulsifying power Methyl oleate
(SA-BD100) 11-62-9 4B △ ○ In case of incorporation treatment
Emulsifying power rise Castor oil, ethoxylated
(LC50D) 4B 4B △ ○ In case of incorporation treatment
Emulsifying power rise polyoxyethylene dodecyl mono ether (LE-5) 9002-92-0 4B × △ Weak emulsifying power polyoxyethylene methyloleate
(OA-1019) 9004-96-0 4B × × Weak emulsifying power Sorbitan monooleate 1338-43-8 4B × △ Weak emulsifying power Methyl searate 112-61-8 3 △ ○ EPA Level 3

- Results: AG-PLG showed the highest emulsifying power of Oregano oil, and showed excellent emulsifying power with addition of 20% or more. Also, it showed constant emulsifying power even when adding more than 20% of ethoxylated and methyl oleate with Castor oil. As a result of the emulsification with the ratio shown in Table 8, the emulsifying performance was also excellent even in the dilution use.

division Emulsifier Input ratio (%) 60% oil blending Oregano oil 60 AG-PLG 10-20 Castor oil, ethoxylated 5 to 10 Methyl oleate 5 to 10

(2) Diffusing agent selection and diffusivity evaluation under various acidity conditions

- Two types of polyalkyleneoxide modified heptamethyltrisiloxane, which has been most widely used in agriculture and proved effective through previous researches, were preferentially selected. As the pH of the selected effect enhancer and microorganism mixture is about 3.5, the contact and diffusibility may be deteriorated depending on the acidity, so that the selection of the dispersing agent is preferred under various acidity conditions.

- Test sample: Microbial culture solution

- Treatment Solution: The above mixed solution was adjusted with citric acid, NaOH added pH 3, pH 7, pH 10

Addition of 0.05% concentration of PMH1 / PMH2 in each pH adjustment liquid

- Test method: Contacting / diffusivity check after injection of 20 ㎕ of 2000 times dilution of each treatments into cucumber and pathogen mycelium

- Test results: As shown in Table 9, although the effect of PMH2 (effective in the range of pH 2 to 10) was negligible, the effect was maintained in a wide range of pH, and PMH2 was selected. 0.0125% or more was added to the pathogenic mycelium to have good diffusion and contact effects. Therefore, it was decided to add more than 6% of the dispersant in the product based on the economic treatment concentration of 500 times or more.

division EPA List 0.05% pH3 pH7 pH 10 contact diffusion contact diffusion contact diffusion polyalkyleneoxide modified heptamethyltrisiloxane (PMH1) 4B + + +++ +++ + + polyalkyleneoxide modified heptamethyltrisiloxane (PMH2) 4B ++ ++ ++ ++ ++ ++

(3) Selection of liquid formulation stabilizer Ⅰ

In the case of microbial liquids using the culture medium, it is necessary to add a certain stabilizer because there is a possibility of degeneration of effective microorganisms due to lack of stability during distribution and deodorization such as odor due to external pollution. Therefore, stability was confirmed by using a stable food additive which can be used as a microbial stabilizer.

- Test sample: Microbial culture solution 3.2 * 107 cfu / ml

- Treatment: 2% of citric acid, 0.1% of sodium silicate, 0.1% of potassium silicate, 0.1% of sodium benzoate, 10% of glucose,

- Investigation: stability after 4 weeks after treatment with stabilizer in culture (microbial density, contamination rate, odor,

Results: As shown in Table 10, microbial safety and physical properties were good in the concentration range of citric acid 2% and Na-benzoate 0.1%. In the case of cultivated dyes, the amount of the two treatments was small, but it increased with time, necessitating the examination of dispersant and suspending agent. Thus, stabilizers were determined to contain 2% citric acid and 0.1% Na-benzoate in the product.

Microbial stability Input ratio
(%) Microbial safety
(cfu / ml) Contamination rate Physical safety Remarks Presence of odor Tint
Occur pH adjustment Citric acid 2 1.0 * 10 ⁷ No No Little Sodium silicate 0.1 1.4 * 10 ⁷ No Yes Many food
Preservative P-sorbate 0.1 0.9 * 10 ⁷ No Yes Little Na-benzoate 0.1 1.7 * 10 ⁷ No No Little Osmotic pressure Glucose 10 2.0 * 10 ⁶ Yes Yes Many

(4) Selection of liquid formulation stabilizer II

In general, culture tiles are fine grains, and they have a specific gravity higher than that of water. Therefore, a stable suspension, a dispersant (surfactant) and a viscosity increasing agent (xanthan gum, kaolin etc.) were added to ensure layer separation and stability.

- Test sample: Microbial culture solution 3.2 * 107 cfu / ml

- Treatment: culture + kaolin 5% + NK-XG 0.5%

Culture + Kaolin 5% + NK-XG 0.5% + UT-100PX 1% or UT-34TDP 2%

Culture medium + Kaolin 5% + NK-XG 0.5% + UT-100PX 1% + SA-SC500 5%

- Investigation: Observation of sedimentation and denaturation after 1 week of manufacturing

- Test results: In the case of kaolin and thickener (NK-XG) treated to inhibit the sedimentation of the tile, sedimentation started from 1 week and the addition of dispersants (UT-100PX and UT-34TDP) The sedimentation rate was slightly slower, but separation and sedimentation occurred gradually. In the case of SA-SC500 additive, which is a liquid suspending agent, no layer separation phenomenon was observed and physical stability was secured, and the final formulation was determined as shown in Table 11 below.

Emulsifier Emulsifier Input ratio (%) Remarks Active ingredient Effective microbial culture 75 to 90 - Thickener NK-XG (xanthan gum) 0.1 to 0.25 Floating tile
fixation Electrodeposition / diffusion UT-100PX 1-3 diffusion UT-34TDP 1-3 diffusion Fixing agent china clay 5-20 Floating tile
fixation Suspending agent SA-SC500 5 Floating tile Total 100

C) Hydration agent

(1) Selection of extender for moisture absorption

- Sample: Zeolite, kaolin, starch, diatomaceous earth

- As shown in Table 12 below, zeolite had high particle size and specific gravity, so it was easily separated from the recovered spores upon dilution. In the case of starch, the specific gravity was too low to float and it was not mixed with water.

- Kaolin was finally selected and used as an extender.

Moisture absorber Hygroscopic Particle Size / Specific Gravity Spore separation phenomenon during hydration Zeolite Inadequate 300mesh / 2.2 Yes china clay Good 325mesh / 2.6 No Starch Good 300mesh / 1.65 Decrease of dispersion power Diatomaceous earth Good 200mesh / 2.1 No

(2) Selection of powdery water-borne surfactants

- 3% of SA-SLS, UT-100PX and UT-425DW, which are widely used for agricultural purposes, were mixed with 5% each of kaolin powder and recovered RDH 1578 spore powder obtained from the above results, Respectively. As shown in Table 13, UT-100PX and UT-425DW were more easily precipitated because of their high specific gravity, and their dispersibility at 3% It is possible to apply it to

Surfactants Dispersibility Remarks SA-SLS 5% Good Easily settled due to high weight UT-100PX 5% Good - UT-425DW 5% Good - UT-100PX (3%) + UT-425DW (3%) Great Good dispersibility even at high concentration dilution

Example  1 Suspension )

Firstly, 20% of AG-PLG, 10% of castor oil, 10% of ethoxylated and 10% of methyl oleate were mixed with 60% of oregano oil at a W / W ratio for 30 minutes to prepare a first emulsion. % Of Caster oil, 2.5% of SA-SC500, 2.5% of SA-BD100, and a mixture of water and water. And 5% of polyalkyleneoxide modified heptamethyltrisiloxane were mixed and stirred for 1 hour to prepare an oily suspension. The ratio of the materials used is shown in Table 15 below.

division Raw material  persons input ratio(%) Active ingredient Effective microbial culture 45% For enhancing effect
additive 60% Oregano oil 25% polyalkyleneoxide modified
heptamethyltrisiloxane (PMH2) 5% Castor oil * 20% For formulation
additive SA-SC500 2.5% SA-BD100 2.5% total

Example  2( Liquid )

NK-XG (Zanthan gum) was prepared by mixing and stirring firstly in 2% of UT-34TDP, a liquid electrodeposition spreader, because it did not easily melt in water at room temperature and had a lot of aggregation phenomenon. Kaolin, which is a sticking extender, and UT-100PX, a powdery surfactant, were mixed and stirred in 75% of the effective microbial culture solution for 30 minutes to prepare a hydrated liquid. Then, NK-XG was mixed with UT-34TDP in small amounts while stirring. 2% of SA-SC500 was finally added and mixed and stirred for 1 hour to prepare a prototype solution. The ratios of the materials used are shown in Table 16 below.

division Raw material name Input ratio (%) Remarks Active ingredient Effective microbial culture 75 - Thickener NK-XG (xanthan gum) 0.1 Floating tile
fixation Electrodeposition / diffusion UT-100PX One diffusion UT-34TDP 2 diffusion Fixing agent china clay 20 Floating tile
fixation Suspending agent SA-SC500 2 Floating tile Total 100

Experimental Example  One

In order to investigate the microbial control effect of each microorganism against cucumber powdery mildew, cucumber (varieties; The microorganisms of Table 17 were mixed as in Example 1 and sprayed three times at 7-day intervals to sufficiently wet the plants. 5 days after the final drug treatment, the diseased area was calculated and the disease severity was calculated. (BX1) + (CX2) + (CX2) +: C, 20.140%: D, and> 40.1% DX3) + (EX4)] / irradiated leaf number X4] X100. Based on the degree of disease, the control group was obtained.

Processing contents Number of microorganisms (cfu / mL) Treatment concentration Natural oil Processing interval and frequency Streptomyces sp. KJA-2 2.0 x 10 ⁸ 250, 500, 1,000 times oregano oil 7 days, 3 times Streptomyces acidiscabies KJA-30 1.2 × 10 ⁸ 〃 〃 〃 Streptomyces tsukionensis JH-19 3.5 × 10 ⁸ 〃 〃 〃 Bacillus subtilis 1.0 × 10 ⁹ 〃 〃 Control drug (product name: Top seed) 200 times 〃 Untreated (water) 〃

As shown in FIG. 22, the higher the treatment concentration, the lower the incidence of the disease, as compared to the four microorganisms such as Streptomyces sp. And Bacillus subtilis which are effective against cucumber powdery mildew. Disease incidence was less than that of the control agent at the treatment concentration of 500 times and 250 times.

As shown in FIG. 23, when the concentration of the microorganism was 500 times, the microorganism alone or the mixture of the microorganism and the natural oil was treated, and the degree of disease was significantly lower in the microorganism and natural oil mixture than the microorganism alone treatment.

As shown in FIG. 24, microorganisms or microbial and natural oil mixtures were treated with dilution of the microorganism or natural oil mixture at the facility house packaging, respectively. As a result, as shown in FIG. 24, .

Experimental Example  2

In order to investigate the effect of microbial control on cucumber powdery mildew, two cucumbers were cultivated (Table 18). In the first packaging test, the microbial agent prepared by the method of Example 1 was sprayed from January 8, and the lesion area was examined 5 days after the final spraying. The secondary packaging test was performed on February 12, and the area of the lesion was examined 5 days after the final spraying. The microorganism of Example 1 selected in the first year was sprayed 4 times, 3 times, and 2 times at 5, 7, and 10 days intervals, respectively, so as to sufficiently wet the plant by adjusting the treatment concentration as shown in Table 19. The disease severity was calculated by examining the lesion area ratio. (B × 1) + (C × 1), and the incidence of disease was calculated as follows: A, 0.15%: B, 5.120% 2) + (D × 3) + (E × 4)] / irradiated leaf × 4] × 100. Based on the degree of disease, the control group was obtained.

Experimental packaging was designed with 3 repetition methods in the facility house (2 packages) in Naju City, Jeollanam - do, and cucumber was treated for 20 weeks or more per repetition. Statistical analysis was performed using ANOVA analysis and Duncan's multiple range test using the R Project program version 3.0.1 (2013).

Crop (breed) How to grow Planting distance Regular machine Fertility (kg / 10a) Remarks Cucumber Autumn Cultivation 120 x 18 cm 10. 5. N-P-K = 19.7? 10.3? 12.2 Primary
(Naju) Cucumber Autumn Cultivation 120 x 18 cm 11. 6 N-P-K = 19.7? 10.3? 12.2 Secondary
(Naju)

Processing contents Number of microorganisms (cfu / mL) Treatment concentration Supplements Processing interval (number of times) Microbial agent
( Bacillus subtilis And
Supplements) 2.3 × 10 ⁹ 250, 500, 1000 times oregano oil, 15% 5 (4), 7 (3), 10 days (twice) Control drug (product name: Top seed) 200 times - 7 days (3 times) Untreated (water)

In the first packaging test, the microbial agent (Bacillus subtilis and adjuvant) of Example 1, which had the effect of controlling cucumber powdery mildew disease, was treated at different treatment concentrations (Table 20). Treatment with 500 and 250 dilution treatments at 5 and 7 days showed significantly higher control than 10 days treatment.

When the concentration of microbial treatment was 500 times and treated at intervals of 7 days, the control effect was 86.2%, which is superior to the control drug.

division Dilution factor Processing interval
(time) Outbreak rate (%) valence
(DMRT) The control
(%) Ⅰ Ⅱ Ⅲ Average Microbial agent 250 times 5 3.0 5.0 5.0 4.3 a 93.2 7 3.0 6.3 4.0 4.4 a 93.0 10 13.0 21.0 14.0 16.0 b 74.6 500 times 5 7.0 9.0 8.0 8.0 a 87.3 7 6.0 11.0 9.0 8.7 a 86.2 10 20.0 25.0 21.0 22.0 c 65.1 1000 times 5 16.0 26.0 20.0 20.7 bc 67.1 7 20.0 32.0 25.0 25.7 c 59.2 10 30.0 41.0 36.0 35.7 d 43.3 Contrast agent 200 times 7 28.0 42.0 34.0 34.7 d 44.9 No treatment water 7 55.0 75.0 59.0 63.0 e

In the second packaging test, the microbial agent (Bacillus subtilis and adjuvant) of Example 1, which had the effect of controlling cucumber powdery mildew disease, was treated at different treatment concentrations (Table 21). The higher the microbial treatment concentration, the higher the control effect. There was a significant difference in the control effect between treatments at 250 and 500 times dilution treatments at 5 days and 7 days intervals and at 500 times 10 days intervals. Compared with the control drug, the treatments were effective at 500 - fold intervals at 7 - day intervals.

division Dilution factor Processing interval
(time) Outbreak rate (%) valence
(DMRT) The control
(%) Ⅰ Ⅱ Ⅲ Average Microbial agent 250 times 5 20.0 12.5 16.7 16.4 a 77.3 7 21.7 18.3 20.0 20.0 ab 72.3 10 35.0 25.0 15.0 25.0 b 65.4 500 times 5 23.3 18.3 21.7 21.1 ab 70.8 7 35.0 23.3 23.3 27.2 b 62.3 10 46.7 33.3 28.3 36.1 c 50.0 1000 times 5 41.7 31.7 31.7 35.0 c 51.5 7 43.3 40.0 31.7 38.3 c 46.9 10 40.0 38.3 41.7 40.0 c 44.6 Contrast agent 200 times 7 43.3 35.0 35.0 37.8 c 47.7 No treatment water 7 80.0 63.3 73.3 72.2 d -

Experimental Example  3

To investigate the effect of microbial treatment on strawberry gray mold disease, strawberry (variety: The microbial agents were identified as Streptomyces acidiscabies were mixed according to the method of Example 1, and then sprayed to the plants three times at 5-day intervals with a dilution factor of 500 times. Disease severity was assessed at 5 days after the final treatment.

Experimental packaging was designed with 3 repetition method in the facility house of Naju city, Jeollanam - do, and strawberry was processed for more than 20 weeks.

Processing contents Number of microorganisms (cfu / mL) Treatment concentration Natural oil Processing interval and frequency Streptomyces acidiscabies 3.8 × 10 ⁶ 500 times oregano oil Every 5 days, 3 times Streptomyces tsukionensis 1.1 × 10 ⁷ 〃 〃 〃 Control drug (product name: Serenade) 500 times 〃 Untreated (water) 〃

Streptomyces with effect on strawberry gray mold After a treatment with different concentrations for 2 acidiscabies microbial species (Fig. 25), a low tendency onset gwayul compared to non-treatment of the selected microorganism or contrast agent and a significant difference was not observed.

Experimental Example  4

To investigate the microbial control effect on strawberry gray mold disease, a test was conducted on strawberries (varieties: Sullivan) for two times (once: two fireflies, twice: three fireflies) ). The microbial agents prepared by the method of Example 1 selected in the first year were sprayed 5 times, 3 times and 2 times at intervals of 3, 5 and 7 days, respectively, so as to sufficiently wet the plants by adjusting the treatment concentration in Table 24. The control effect was investigated at 5 days after the final treatment. The control was calculated as [(onset rate of untreated onset rate - incidence rate of treatments) / onset rate of untreated onset] × 100.

Experimental packaging was designed with 3 repetition method in the facility house of Naju city, Jeollanam - do, and strawberry was processed for more than 20 weeks. Statistical analysis was performed using ANOVA analysis and Duncan's multiple range test using the R Project program version 3.0.1 (2013).

Crop (breed) How to grow Planting distance Full day Fertility (kg / 10a) Remarks Strawberry
(Shuli) Facility cultivation 120 x 15 cm 10. 26 N-P-K: 9.6? 4.9? 7.4 Naju 2 Painting Room
(March 4) Strawberry
(Shuli) Facility cultivation 120 x 15 cm 10. 26 N-P-K: 9.6? 4.9? 7.4 Naju
3 air conditioner
(March 28)

Processing contents Number of microorganisms
(cfu / mL) Treatment concentration Supplements Processing interval and frequency Microbiology ( Streptomyces
acidiscabies And adjuvants) 1.0 × 10 ⁸ 250, 500 times oregano oil, 15% 3 (5), 5 (3), 7 days (twice) Control drug (product name: Serenade) 500 times 7 days (twice) No treatment water

In order to investigate the effect of the microbial agent on the control of the strawberry gray mold of the first year of the first packaging test in Example 1, the microbial treatment concentration was varied at the early stage of March of the strawberry 2 festival (Table 25) And the incidence of disease was low. The treatments with 250 times of treatment concentration at 35 days interval and 500 times of 3 days interval treatment showed the same control effect as the control drug.

division Dilution factor Processing interval
(time) Onset rate (%) valence
(DMRT) The control
(%) Ⅰ Ⅱ Ⅲ Average Microbial agent 250 3 42.1 28.7 16.4 29.1 ab 53.7 5 32.0 30.4 27.0 29.8 ab 52.5 7 39.6 35.6 27.6 34.3 bc 45.4 500 3 32.7 29.3 21.6 27.9 ab 55.6 5 41.9 37.8 34.6 38.1 c 39.3 7 47.0 42.7 31.5 40.4 c 35.6 Contrast agent 500 7 28.6 20.0 18.9 22.5 a 64.2 No treatment water 7 66.9 61.0 60.3 62.8 d -

For the second packaging test, the microbial agent of Example 1 of the first year was treated at the end of March, which is the strawberry 3 flower, in the same manner as the first packaging test (Table 26). . The treatments with 250 times of 35 - day intervals showed the same level of control as the control. Therefore, it is recommended that the microbial agent be diluted 250 times and treated at intervals of 5 days in order to show the same effect as the control agent.

division Dilution factor Processing interval
(time) Onset rate (%) valence
(DMRT) The control
(%) Ⅰ Ⅱ Ⅲ Average Microbial agent 250 3 31.1 23.4 28.3 27.6 a 54.2 5 32.9 22.3 29.0 28.1 ab 53.3 7 37.3 24.6 32.4 31.5 c 47.7 500 3 34.4 26.4 31.6 30.8 bc 48.8 5 38.4 29.9 36.9 35.1 d 41.6 7 39.9 33.6 37.4 37.0 d 38.5 Contrast agent 500 7 30.7 22.2 23.2 25.4 a 57.8 No treatment water 7 62.1 58.4 59.8 60.1 e -

The preferred embodiments of the present invention have been described in detail above. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Accordingly, the scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning, range, and equivalence of the claims are included in the scope of the present invention Should be interpreted.

Claims (5)

Streptomyces xanthocidicus Natural herbicide containing the culture medium of JH-19 as an active ingredient and having an active activity against cucumber powdery mildew.
The method according to claim 1,
Oregano oil, polyalkylene oxide modified heptamethyltrisiloxane (PMH2) and castor oil were added as additives for improving the effect.
A natural disinfectant characterized by further comprising a surfactant as an additive for formulation.
delete The method according to claim 1,
A surfactant as a dispersing agent, and a kaolin as a fixing agent.
delete

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