WO2014193162A1

WO2014193162A1 - Preparation method for biopesticide using paecilomyces sp. strain

Info

Publication number: WO2014193162A1
Application number: PCT/KR2014/004766
Authority: WO
Inventors: 박호용; 손광희; 전기; 신동하; 오현우; 조한영
Original assignee: 한국생명공학연구원
Priority date: 2013-05-31
Filing date: 2014-05-28
Publication date: 2014-12-04
Also published as: JP2016526043A; KR20140141857A; JP6387438B2; JP2017197550A

Abstract

The present invention relates to a preparation method for an environment-friendly organic material, or a biopesticide using a Paecilomyces sp. strain. More particularly, a water-dispersible powder, emulsifiable concentrate or oil agent composition for pest control, which are prepared by mixing, in an optimum condition, an active spore produced from the Paecilomyces sp. strain with an excipient, emulsifier, etc., is easily dispersed, suspended or blended into water or an organic solvent, maintains a unique insect prevention effect, increases stability, and thus can be effectively used for pest control.

Description

【Specification】

[Name of invention]

Manufacturing method of biological pesticide using strain of genus Pacciomyces 【Technical field】

The present invention relates to a method for preparing a pesticide biopesticide using active spores of the strains of the genus Pacylomyces (/ ¾ec / AM? Yces).

Background Art

Biopesticides, also called biocontrol agents or green pesticides, use pathogens, pests, and pests that cause damage to crop production and preservation using materials that exist in nature and materials derived therefrom. Crop supplements that control weeds. The US Environmental Protection Agency (US EPA) classifies biochemical pesticides derived from natural products, microbial pesticides using pest control microorganisms, and piant-incorporated protectants (PIPs). EU refers to pesticides using microorganisms or natural products. In Korea, the same or similar concept as biopesticides is used to control crop pests without artificial chemical treatment of the crop protection function that natural biological resources (plants, microorganisms, insects, etc.) are unique. It is classified as a natural crop protection material.

Crop protection materials are classified into biopesticides and chemical pesticides, and biopesticides are classified into biological pesticides, including microbiological pesticides and biochemical pesticides, and others that use natural enemies, ships, and parasites. Among these biopesticides, using microorganisms, asbestos, etc., are classified as organic materials.

Of agriculture that has been around for centuries. The method was developed with the aim of mass production and consumption. In addition to well-being demands, food safety is emphasized above all. Therefore, the demand for organic produce is rapidly increasing, and the growing area is continuously increasing. In particular, since the food safety problem is linked to the primary safety of the crops produced, the government is developing a plan to promote eco-friendly agriculture, while increasing the share of eco-friendly agricultural products and continuously reducing the use of chemical pesticides and fertilizers at the current level. I'm working on it.

In order to secure the international competitiveness of China's agricultural industry, the production of environment-friendly and safe agricultural products is inevitable, and for this purpose, the development and dissemination of excellent biological pesticides that can replace chemical pesticides is essential. This is essential for the 10% increase in the production of eco-friendly agricultural products and the 40% reduction in the use of chemical pesticides by 2013.

Currently, there are about 370 biopesticides registered around the world. Among them, 270 species of pesticides are used, accounting for about 75% of the total, and the development of biopesticides is based on relatively small companies. Development of microbial pesticides based on microorganisms is a major feature. Looking at the types of microorganisms registered and used as microbial pesticides, there are the most bears and bacteria, followed by viruses used for pests. Pest control using entomopathogenic fungi is an environmentally friendly control means, selectively acting only on target pests, inhibiting the emergence of pest resistance, enabling continuous control, and not at all in the environment, humans and plants. It has many advantages, such as the ability to produce high-quality agricultural products without harm and no residual toxicity. Therefore, many agricultural developed countries including the United States have separated, mass-produced and formulated insect pathogenic bears suitable for their environment on a national level. In practical use It is true.

Even in Korea, the research and development to date is limited to the development of insect pathogenic fungi probiotics, making it difficult to secure industrial performance. In order to apply probiotics to agricultural sites, it is necessary to ensure the stability of the formulation through formulation to maintain their activity. However, due to the lack of technology related to it, it is difficult to secure the economic efficiency of the developed product and the excellent effect in the field, and these problems are a significant cause for the sluggish market expansion. Therefore, the present inventors endeavored to develop a biopesticide that has a pest repellent effect and stability of the formulation through the formulation, as a result of mixing the active spores produced from the strains of the genus Paciromomyces in the optimal conditions with excipients, emulsions, etc. Or as a result of preparation in oil form, wherein the wetting agent, emulsion or oil agent is easily dispersed, suspended or mixed in water or an organic solvent, the inherent pest insect repellent effect is maintained, and the stability is increased, thereby confirming the composition. The present invention has been completed by confirming that it can be usefully used for pest control.

[Detailed Description of the Invention]

[Technical problem]

An object of the present invention is pasilomyase

It is to provide a pesticide biopesticide and a method for producing the same using the active spores of the strain.

Technical Solution

In order to achieve the above object ，

The present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Paciromomyses (/ ¾ec / ¾ /? Yces), 30 to 50 parts by weight of corn starch or potato starch, excipients 30 to Hydrating composition for pest control comprising 50 parts by weight and 30 to 50 parts by weight of texturing to provide.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Pasilomyses, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifier 2 to 5 weights based on the total weight of the composition It provides a pesticide emulsion composition for controlling pests.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, emulsifier 2 to 5 parts by weight of the culture medium of the strain of the genus Pacillomyces relative to the total weight of the oil composition The agent composition is provided.

In addition, the present invention

1) culturing the strain of genus Pacillomyces to obtain a culture solution or active spores of the genus of Paciromomyces; ―

2) 5 to 10 parts by weight of the culture solution or activated spores of the genus of Paciromomyces obtained in step 1) with respect to the total amount of the hydrating agent composition, 30 to 50 parts by weight of corn starch or potato starch, 30 to 50 parts by weight of excipient And it provides a method for producing a pesticide wetting agent composition comprising the step of mixing 30 to 50 parts by weight of texturing.

In addition, the present invention

1) culturing the strain of genus Pacillomyces to obtain a culture solution or active spores of the genus of Paciromomyces;

2) based on the total weight of the emulsion composition, 5 to 10 parts by weight of the culture or active spores of the genus of Paciromomyces obtained in ¾ type 1), 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifier It provides a pest control emulsion preparation method comprising the step of mixing 2 to 5 parts by weight. In addition, the present invention

2) with respect to the total weight of the oil composition, obtained in step 1) Provided is a method for preparing an oil composition for controlling sea layer comprising the step of mixing 5 to 10 parts by weight of the culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, and 2 to 5 parts by weight of an emulsifier. .

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Pasilomyces (/ ec / ¾? Yces), 30 to 50 parts by weight of corn starch or potato starch, relative to the total weight of the hydration composition Provided is a composition for use as a wetting agent for controlling pests comprising a composition comprising 30 to 50 parts by weight and 30 to 50 parts by weight of dextrin.

In addition, the present invention emulsions ^{compositions'} total weight losses indeed My broth or its activated spores process sp 5 to 10 with respect to the weight parts of formula physical properties five days 50 to 60 parts by weight, 30 to 40 parts by weight of water, emulsifier 2-5 It provides a use for using a composition comprising parts by weight as an emulsion for controlling pests.

In addition, the present invention pest control composition comprising the culture solution of the strain of the genus Pacillomyces or 5 to 10 parts by weight of its active spores, 90 to 98 parts by weight of vegetable oil, 2 to 5 parts by weight of the emulsifier relative to the total weight of the oil composition It is intended to be used as a solvent oil.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Paciromomyses (/ ¾ec / / z? Yces), 30 to 50 parts by weight of corn starch or potato starch, Provided is a pest control method comprising the step of treating a plant, soil or seed with a pesticide wetting agent composition comprising 30 to 50 parts by weight of excipients and 30 to 50 parts by weight of dextrin.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsified 쩨 2 to 5 parts by weight of the total amount of emulsion composition Provides a pest control method comprising the step of treating the pesticide composition for pest control comprising a part to plants, soil or seeds.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Pacillomyces relative to the total weight of the oil composition, vegetable oil 90 to 98 It provides a pest control method comprising the step of treating the oil-based composition for controlling the sea layer comprising parts by weight, emulsifier 2 to 5 parts by weight to plants, soil or seeds.

Advantageous Effects

The present invention relates to an insecticide, emulsion, or oil composition for controlling pests prepared by mixing active spores produced from the genus of Pasylomyces (/ ¾ec / ¾7 ces) with an excipient, an emulsifier, and the like under optimum conditions. Easily dispersed, suspended or mixed in water or organic solvents, the inherent pest insect repelling effect is maintained, and stability is increased, so it can be usefully used for pest control.

[Brief Description of Drawings]

1 is a diagram confirming the stability of the excipients for the dispersion, suspension or mixing of the active spores of the Pasylomyces lysinus (/ ¾ec / A¾¾ ces J γ acinus) HY-4 strain. Figure 2 is a diagram showing the spore survival rate (%) of the active spores of the active spores of the Pacillomyces lysinus HY-4 strain containing various emulsions (days).

Figure 3 is a diagram confirming the stability of the active spores of the Pacillomyces lysinus HY-4 strain against the emulsifier.

4 is a diagram confirming the improvement of the stability of the active spores of the Pacillomyces lysinus HY-4 strain against the antioxidant.

Figure 5 is a diagram confirming the pesticidal effect of the hydrating agent and emulsifying agent containing the active spores of the Pacillomyces lysinus HY-4 strain.

[Best form for implementation of the invention]

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail. The present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Paciromomyces (/ ¾ec // OTyces), 30 to 50 parts by weight of corn starch or potato starch, 30 to 50 parts by weight of the excipient composition part, And it provides a pesticide wetting agent composition comprising 30 to 50 parts by weight of dextrin.

The strain of the genus of Pacillomyces is preferably Pasilomyces lysinus ¾ «: / 7iM7yces lilacinus) HY-4 strain deposited with accession number KCTC 0395BP, but is not limited thereto.

The excipient is not toxic to microorganisms, it is preferable to use what is acceptable as an agricultural soil treatment agent, there is no clogging of the nozzle of the sprayer, it is more preferable to be able to control the water content, the most preferred is bentonite, kaolin or zeolite Preferred but not limited to this.

The hydration composition for controlling the marine layer is preferably one which improves pest control activity and stability, but is not limited thereto.

The hydrating agent composition may further include a known adjuvant as necessary. The adjuvant may include, but is not limited to, a stabilizer, an antioxidant, a thickener, and an antifoaming agent.

The culture solution S of the strain of the genus Paciromomyses (/ ¾ec / (¾i ces), when its active spores contain 5 parts by weight or less based on the total weight of the hydrated composition, since the pest control effect is significantly reduced, The culture solution of the genus Seth or its active spores preferably contain 5 to 10 parts by weight based on the total weight of the hydrating agent composition, but is not limited thereto.

When the culture solution or the active spores of the strain of the genus Pacylomyces (/ ¾ec7? Yces) contains 10 parts by weight or more with respect to the total amount of the hydrating agent composition, the contamination is caused by the bears floating in the air, Since the viability of the culture medium of the strains of the genus Myes or the viability of the active spores is significantly reduced, the pest control effect is significantly reduced, so the culture or the active spores of the strains of the genus Paciromomyces is 5 to the total weight of the hydrating agent composition It is preferably one to 10 parts by weight but is not limited thereto.

When the corn starch or potato starch contains 30 parts by weight or less or 50 parts by weight or more with respect to the total weight of the hydration composition, the hydration composition is in water It is preferable to contain 30 to 50 parts by weight based on the total weight of the corn starch or potato starch since the activity of the sea layer control is significantly reduced because it is not significantly mixed or suspended.

The hydrous composition should be specifically designed to have a suitable powder, dispersing power, and water content so as to be in suspension when mixed with water, but when it contains an excipient of 30 parts by weight or less or 50 parts by weight or more based on the total weight of the hydrous composition, Since the germination rate and susceptibility of the activated spores are markedly reduced, the pest control activity is reduced, so that the excipient preferably contains 30 to 50 parts by weight based on the total weight. In a specific embodiment of the present invention, the present inventors separated and identified a microorganism having an insecticidal effect on the sea layer from the soil sample and named it as Pasylomyces lyrecinus HY-4, and deposited it in the Korea Biotechnology Research Institute Gene Bank. (Accession No. KCTC 0395BP).

In addition, the present inventors, after confirming the stability of the excipient for the dispersion, suspension and mixing of the strain active spores of the genus Pacillomyces, after mixing the active spores and excipients of the strains of the genus Pacillomyces, spore germination viable count by storage period As a result, the active spores mixed with bentonite, kaoli or zeolite were confirmed that the germination rate of the active spores was maintained at 70% or more even after storing for 6 months or more (see Fig. 1).

In addition, the present inventors mixed the excipients in the active spores to confirm the suspension of the active spores of the strains of the genus Pacillomyces using the excipients, and measured the susceptibility of each concentration, the total weight of the hydrating agent composition When it contains 30 parts by weight to 50 parts by weight of the excipient, it was confirmed that the suspension properties are remarkably excellent.

In addition, the present inventors, based on the total weight of the hydrating agent composition, 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Pasilomyces (¾ec; 7o / »yces), corn starch or potato starch ₃₀ to 50 parts by weight, and excipients After mixing 30 to 50 parts by weight to prepare a hydration composition, and confirmed the insecticidal effect, Purified activated spores did not show a significant killing effect, while the hydrating composition of the present invention was confirmed to exhibit a pesticidal effect of 50 to 100% (see Fig. 5 and Table 1).

Therefore, the hydrating agent composition of the present invention facilitates the dispersion, mixing and suspending the active spores produced from the strains of the genus Pacillomyces, and can maintain and stabilize the intrinsic pest control activity, useful for controlling pests. Can be used. In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifiers 2 to 5 parts by weight based on the total amount of the emulsifier composition Provided is an emulsion composition for controlling pests.

The strain of the genus of Pacillomyces is preferably a Pasylomyces lysinus (¾ec / / o / »yces lilacinus) HY-4 strain deposited with accession number KCTC 0395BP, but is not limited thereto.

The vegetable oil is preferably at least one selected from the group consisting of corn oil, soybean oil, grape seed oil, canola oil, olive oil, kerosene and animal oil, but is not limited thereto.

The emulsifier is preferably at least one selected from the group consisting of Tween 20, Tween 80, Span 80, Span 85, and Triton X-100, but is not limited thereto.

The pesticide emulsion composition for controlling pests is preferably, but not limited to, improved seaweed control activity and ¾ quality.

The emulsion composition may further include a known adjuvant as necessary. The adjuvant may include, in particular, stabilizers, antioxidants, thickeners and antifoaming agents, but is not limited thereto. The antioxidant is tertiary butyl hydroquinone (TBHQ) 'butyl hydroxyl toluene (BHT), Butylated hydroxy 1 anisole (BHA), propyl gallate (PG), vitamin C vitamin C, Vt.C), vitamin E vitamin E, Vt.E), lecithin or iron oxide ( Fe ₂ 0 ₃ ) is preferably, but not limited to.

The culture solution or the active spores of the strains of the genus Paciromomyces ¾ec / ¾7 ces) containing less than 5 parts by weight relative to the total weight of the emulsion composition, since the pest control effect is significantly reduced, The culture solution of or the active spores thereof is preferably contained 5 to 10 parts by weight based on the total weight of the emulsion composition, but is not limited thereto.

When the culture solution or the active spores of the strain of the genus Pacylomyces (/ ¾ec / / ces) contains 10 parts by weight or more based on the total weight of the emulsion composition, contamination is caused by the fungus suspended in the air, Since the viability of the culture medium or the active spores of the strains of the genus Myes is significantly reduced and the effect of controlling pests is significantly reduced, the culture medium or its active spores of the strains of the genus Paciromomyces is 5 to the total weight of the emulsion composition. It is preferably one to 10 parts by weight but is not limited thereto.

The emulsifier preferably contains 2 to 5 parts by weight in the total weight of the emulsion composition, and when it contains 2 parts by weight or less, the emulsion composition of the present invention is not mixed or suspended in the solvent, and the pest control effect is significantly reduced. In a specific embodiment of the present invention, the present inventors suspended the active spores of the present invention in corn oil, soybean oil, etc., in order to examine the stability of the active spores for various emulsions, and then check the germination of spores to measure the survival rate. As a result, it was confirmed that the emulsion such as corn oil does not affect the germination of spores (see Fig. 2).

In addition, the present inventors, in order to examine the stability of the active spores for various emulsifiers, adding the active spores of the present invention to soybean oil, suspended by the addition of emulsifiers and then confirmed the germination of the spores, and as a result of measuring the survival rate, The emulsifier It was confirmed that there was no significant effect on the germination of spores until 6 months (see FIG. 2).

In addition, the present inventors confirmed the stability improvement effect of the active spores of the present invention using various antioxidants, it was confirmed that the active spores with the addition of antioxidants maintained the spore survival rate of 88% or more even after 210 days (see Fig. 4) .

Therefore, the emulsion composition of the present invention facilitates dispersing, mixing and suspending the active spores produced from the strains of the genus Pacillomyces in water and solvents, and can maintain and stabilize the intrinsic pest control activity. It can be usefully used. In addition, the present invention is a pest control oil comprising a culture solution of the genus of Pacillomyces strain or 5 to 10 parts by weight of its active spores, 90 to 98 parts by weight of vegetable oil, 2 to 5 parts by weight of an emulsifier based on the total weight of the oil composition The agent composition is provided.

The strain of the genus of Pacillomyces is preferably Pasilomyces lysinus 0¾ec / Myces lilacinus) HY-4 strain deposited with accession number KCTC 0395BP, but is not limited thereto.

The vegetable oil is preferably at least one selected from the group consisting of corn oil, soybean oil, grape seed oil, canola oil, olive oil kerosene and animal oil, but is not limited thereto.

The oil-based composition for controlling the sea layer is preferably an improved sea layer control activity and stability, but is not limited thereto.

The oil composition may further include a known adjuvant as necessary. The adjuvant may include, but is not limited to, stabilizers, antioxidants, thickeners and antifoams. The antioxidant Tertiary-butylhydroquinone (TBHQ), butyl hydroxy 1 toluene (BHT), butylated hydroxy 1 anisole (BHA), propyl gal late (PG), Vitamin C (vitamin C, Vt.C), vitamin EOitamin E, Vt.E), lecithin (lecithin) or iron oxide (Fe ₂ 0 ₃ ) is preferably, but not limited to.

The culture solution or the active spores of the strain of the genus Paciromomyses (/ ¾ec / / o¾rces) contains 5 parts by weight or less based on the total weight of the oil composition, so the pest control effect is significantly reduced, Culture medium of the genus strain or active spores thereof is preferably contained 5 to 10 parts by weight based on the total weight of the emulsion composition, but is not limited thereto.

When the culture solution or the active spores of the strain of the genus Paciromomyses (/ ¾ec / / o / ¾ ces) contains 10 parts by weight or more based on the total weight of the emulsion composition, contamination is caused by the bears floating in the air ， The viability of the culture of the strain of the genus Paciromius or its active spores are significantly reduced, so that the pest control effect is significantly reduced, the culture of the strain of the genus of Paciromomyces or its active spores total emulsion composition It is preferable to contain 5 to 10 parts by weight based on the weight, but is not limited thereto.

The emulsifier preferably contains 2 to 5 parts by weight of the total weight of the oil composition, and when it contains 2 parts by weight or less, the oil composition of the present invention is not mixed or suspended in the solvent, and the pest control effect is significantly reduced. .

Therefore, the oil-based composition of the present invention facilitates dispersing, mixing and suspending the active spores produced from the strains of the genus Pacillomyces in the solvent, and can be useful for controlling pests, since they can maintain and stabilize their own marine control activities. Can be used. In addition, the present invention provides a method for preparing a hydrated emulsion emulsion or oil composition for controlling the sea layer using a culture solution or active spores of the genus of Pacillomyces to provide.

The hydrating agent composition is preferably prepared by the following method, but is not limited thereto:

2) 5 to 10 parts by weight of the culture solution or active spores of the _Pacillomyces strain obtained in step 1), 30 to 50 parts by weight of corn starch or potato starch, 30 to 50 parts by weight of excipient, Parts, and 30-50 parts by weight of dextrin.

The emulsion composition is preferably prepared by the following method, but is not limited thereto:

1) culturing the strain of genus Pacillomyces to obtain a culture medium or activated spores of the genus of Paciromomyces;

2) Regarding the total weight of the emulsion composition, 5 to 10 parts by weight of the culture solution or activated spores of the strain of genus Pacillomyces obtained in step 1), 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifier 2 Mixing 5 parts by weight to 5 parts by weight.

The oil composition is preferably prepared by the following method, but is not limited thereto:

2) 5 to 10 parts by weight of the culture medium or active spores thereof, 9 to 98 parts by weight of vegetable oil, 2 to 5 parts by weight of emulsifier Step of combining. The strain of the genus of Pacillomyces is preferably one of the Pasylomyces lysinus (/ ¾ec / gs Ul acinus) HY-4 strain deposited with accession number KCTC 0395BP, but is not limited thereto.

The composition prepared in step 2) is a known adjuvant as needed It may further include. The adjuvant may include, in particular, stabilizers, antioxidants, thickeners and antifoaming agents, but is not limited thereto.

Therefore, the hydrate, emulsion or oil composition prepared by the method of the present invention facilitates the dispersion, mixing and suspending the active spores produced from the strains of the genus Pacylomyces in water or solvents, and provides unique pest control activity. Since it can maintain and stabilize, it can be usefully used for pest control. In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores of the strain of the genus Paciromomyces (/ ¾ecJo / z7yces), corn starch or potato starch 30 to 50 parts by weight, excipient 30 to 50 relative to the total weight of the hydrating agent composition A composition comprising 30 parts by weight and 30-50 parts by weight of texturing is provided for use as a wetting agent for controlling pests.

In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, and 2 to 5 parts by weight of an emulsifier based on the total amount of the emulsion composition. It provides a use for using the composition comprising as an emulsion for controlling pests.

In addition, the present invention is a pest control composition containing 5 to 10 parts by weight of the culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, 2 to 5 parts by weight of the emulsifier based on the total weight of the oil composition Provided for use as a solvent oil.

Accordingly, the hydrating composition, emulsion composition and oil composition of the present invention facilitate the dispersion, mixing and suspending the active spores produced from the strains of the genus Pacillomyces in water, and can maintain and stabilize the unique pest control activity. Therefore, it can be usefully used for the purpose of using as a pesticide wetting agent, emulsion and oil agent. In addition, the present invention is based on the total weight of the hydrating agent composition of the culture solution of the strains of the genus Paciromomyses (/ ¾ec / o / ¾ ces) or its active spores 5 to 10 30 to 50 parts by weight of corn starch or potato starch, 30 to 50 parts by weight of excipients and 30 to 50 parts by weight of dextrin 30 to 50 parts by weight of the pesticide control method comprising the step of treating the plant, soil or seed to the hydration control composition To provide. In addition, the present invention is 5 to 10 parts by weight of the culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifiers 2 to 5 parts by weight based on the total weight of the emulsion composition It provides a pest control method comprising the step of treating the pesticide emulsion composition for controlling plants, soil or seeds.

In addition, the present invention is a seawater control oil comprising a culture solution of the strain of the genus Pacillomyces or 5 to 10 parts by weight of its active spores, 90 to 98 parts by weight of vegetable oil, 2 to 5 parts by weight of an emulsifier based on the total weight of the oil composition Provided by a pest control method comprising the step of treating the composition to plants, soil or seeds. The composition is preferably used in the liquid state irrigation to the plant, immersed or sprayed on the seed of the crop or coated on the seed, but is not limited thereto.

The control method using the above composition for controlling plant pathogens is generally performed in general, that is, spraying (for example, spraying, misting, atomizing, powder spraying, granulating spraying, sleeping, permanent use, etc.), soil application (for example, For example, it may be performed by shaking, irrigation, etc.), surface use (for example, coating, smearing, coating, etc.), dipping, and smoking. The amount can be appropriately determined according to the damage situation, the method of application, and the place of application. Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely illustrative of the present invention, the contents of the present invention is not limited to the following examples.

Example 1 Pre-cultivation of the Pacillomyces lysinus HY-4 Strain

Pacillomyces lysinus HY-4 strain (Accession No. KCTC 0395BP; PDA (Potato Dextrose Agar) medium was used for the spawn culture for storage and culture of the Republic of Korea registration number: 10-0475131. Spawn culture was incubated in Petri dish (87 χ 15 mm) in PDA medium at 28 ⁰ C, humidity 65% in dark conditions for 7 days.

Example 2 Cultivation of Pacillomyces lysinus HY-4 Strain

The maximum mycelium was made in the culture medium through the liquid culture as the primary culture for the culture of the Pacillomyces lysinus HY-4 strain, and then the secondary culture was performed as a secondary culture for the preparation of the pesticide. Specifically, the primary liquid culture for mycelium culture was incubated for 7 days at 28 ° C, 180 rpm stirring conditions in PBD medium. After that, to Petri dish (150 χ 25 隨)

25 Czapek medium conditions were made, and the primary culture solution formed with 2 ιι mycelium was dispensed and spread evenly with a spreader, and solid culture was performed for 20 days.

<Example 3> Spore recovery of strain Pacillomyces lysinus HY-4

In the Petri dish of <Example 2> it was dried for 10 days so that the medium is completely dried in Humb hood (Hume hood) for spore recovery of the Pacillomyces lysinus HY-4 strain formed through 7 days solid culture. Spores were recovered through a scraper in the dried Petri dish culture medium. Example 4 Confirmation of stability of excipients for dispersion, suspension and mixing of active spores

The stability of the excipients was confirmed using the active spores of the Pacillomyces Lara sinus HY-4 strain recovered in Example 3.

Specifically, the number of spore germination microorganisms was measured using the minerals including bentonite, kaolin, zeolite, etc., which are mainly used as excipients of bio pesticides or eco-friendly organic farming materials, and polysaccharides such as dextrin, corn starch and rice bran. . 100 g of spores and excipients are mixed in l: 9 (w: w) The survival rate of the spores was measured every 30 days while storing 250 bottles (Amber bottle, SPL) containing the mixture at room temperature. The survival rate of spores was 0.5 g of the sample suspended in 4.5 saline, and diluted in steps of 1/10 of each dilution solution.

0.1 was evenly spread on PDA flat broth, incubated at 28 ° C. for 2 days, and colonies were measured.

As a result, as shown in FIG. 1, it was confirmed that the spore germination rate of each excipient changed from the storage period of 4 months. In particular, after 6 months, the spore germination of the sample containing no excipients was reduced to 50% or less, while the active spores containing the excipients were stored up to 11 months, but the germination rate of spores was maintained at 50% or more (Fig. One).

<Example 5> Suspension of activated spores using excipients

The susceptibility of the active spores of the Pacillomyces lysinus HY-4 strain recovered in <Example 3> for the excipient was confirmed by the wetting time. Specifically, minerals such as bentonite, kaolin, and zeolite, and polysaccharides such as dextrin, corn starch, rice bran, and the like were mixed to measure suspension at each concentration. The ratio of spores and excipients is mixed from l: 9 (w: w) to 10: 0 (w: w), respectively, and weighed by mixing the LG composition in a conical tube (SPL) containing 50πι water. After standing at room temperature for 30 minutes, the supernatant of 40 was precisely removed with a pipette, and the remaining suspension was dried to measure a dry weight (W2).

Suspension (%, w / w) was calculated by the following equation.二 / ^ 、 M

W \ 9

As a result, a composition comprising 10% (w / w) spores and 9 (w / w) minerals had a susceptibility of 85-92¾> (w / w), 10% (w / w) spores, 90% (w). / w) compositions comprising polysaccharides contain 25-35% (w / w) suspension, 10% (w / w) spores, 45% polysaccharides, 45% (w / w) minerals In the composition, a suspension of 70 to 75% (w / w) was confirmed.

Example 6 Stability of Active Spores for Various Oils

The stability of various emulsions using the active spores of the Pacillomyces lysinus HY-4 strain recovered in <Example 3> was confirmed.

Specifically, the active spores (w / v) of the Pacillomyces lysinus HY-4 strains were suspended in various vegetable oils such as corn oil, soybean oil, grape seed oil, canola oil, and olive oil, and then stored at room temperature. Each storage sample was checked for the survival of spores every 30 days for 180 days. The viability of the spores was determined by suspending 0.5 g of the sample in 4.5 saline, diluting stepwise in 1/10 units, and uniformly diluting 0.1 m of the diluent in the PDA plate medium for 2 days at 28 ° C. After incubation, colonies were measured.

As a result, as shown in Fig. 2, the spore survival rate was significantly decreased in the case of the emulsion without an emulsion starting from 120 days after storage, maintaining 5 survival rates after 180 days, while the storage sample used as an emulsion was continuously Spore viability of at least 90% was maintained (FIG. 2).

Example 7 Stability of Active Spores for Various Emulsifiers

The stability of various emulsifiers using the active spores of the Pacillomyces lysinus HY-4 strain recovered in <Example 3> was confirmed.

Specifically, 5% (w / v) of the active spores of the strain of Pacillomyces lysinus HY-4 in soybean oil, Tween 20, Tween 80, Span 80, Span 85 and Triton

X-100 (Sigma-Aldrich Co., LLC.) Each emulsifier was suspended with the addition of 2.5% (w / v) and then stored at silver. Each storage sample was checked for survival by checking the germination of spores every 30 days for 240 days. The survival rate of the spores was 0.5 g of the sample suspended in 4.5 m physiological saline, diluted in steps of 1/10 each, 0.1 dilution was evenly spread on a PDA plate medium, incubated for 2 days at 28 ⁰ C One Next colonies were measured.

As a result, as shown in Figure 3, it was confirmed that the active spores containing a variety of emulsifiers did not show a significant difference in the spore survival rate compared to the control group not treated with the emulsifier (Fig. 3).

<Example 8> Stability improvement of activated spores against various antioxidants using antioxidants

It was confirmed that the stability of various antioxidants using the active spores of the Pacillomyces lysinus HY-4 strain recovered in <Example 3> suspended or coated in vegetable oil was confirmed.

Specifically, after suspending l% (w / v) of the active spores of the strains of Pacillomyces lysinus HY-4 in soybean oil, tertiary-butylhydroquinone (TBHQ) and butyl hydroxyluene (butyl hydroxy 1 toluene, BHT), butylated hydroxy 1 anisole (BHA), propyl gallate (PG), vitamin C vitamin C, Vt.C), vitamin E (vitamin E, Vt.E), lecithin (lecithin) and 0.02% (w / v) of antioxidants such as iron oxide (Fe ₂ O ₃ ) were added and stored at room temperature. Each stored sample was checked for survival for 210 days at 30 day intervals.

As a result, as shown in FIG. 4, the survival rate of spores stored for 180 days was reduced to 50%, while the storage samples to which antioxidants except vitamin C and vitamin E were added maintained 80 or more spore survival rates. In particular, it was confirmed that after 210 days, the storage sample to which butylhydroxyluene was added among the storage samples maintained the spore viability and had the highest stability against spores (FIG. 4). Example 9 Preparation of Insect Repellent Agent Using Pacylomyces spores

For the purpose of the stable formulation of the active spores of the Pacillomyces lysinus HY-4 strain was prepared a hydrating agent used by mixing or suspending in water.

Specifically, as in <Example 4> and <Example 5>, pasilomyces 5-10% (w / v) active spores of lirasinus HY-4 strain, 5-10% (w / v) cornstarch, 30-50% (w / v) bentonite, 30-50% (w / v) dextrin v) was combined to prepare a hydrating composition. The bentonite may be replaced with minerals such as kaolin and zeolite. Corn starch can also be substituted for potato starch.

Example 10 Preparation of Pesticide Emulsifying Agent Using Pacylomyces spores In order to stably formulate active spores of the strain of Pacylomyces lysinus HY-4, an emulsion used by mixing or suspending it in a solvent and water is used. Prepared. Specifically, as in <Example 4> and <Example 6>, 5 to 10% (w / v) of active spores of the strains of Pacillomyces larasinus HY— 4, and 50 to 60 (w / v) of vegetable oils. An emulsion composition was prepared by mixing water, 30-4 C »(w / v), and emulsifier 2-5% (w / v). The vegetable oil may be substituted for kerosene or animal oil which is widely used for agriculture. Emulsifiers can vary the emulsifier component according to polarity such as Tween 20, Tween 80, Span 80, Span 85, Triton X-100.

<Example 11> Preparation of oil for anti-layer control using active spores of the genus of Paciromomyces Oil agent used by mixing or suspending in a solvent for the purpose of stably formulating the active spores of the strains of Pacillomyces lysinus HY-4 Was prepared.

Specifically, as in <Example 4> and <Example 6>, 5 to 10% O / v of spores of the Pacillomyces lysinus HY-4 strain and 90 to 98% (w / v) of vegetable oil , Emulsifier 2-5% (w / v) was used. The vegetable oil may be substituted for kerosene or animal oil which is widely used for agriculture. The emulsifier may vary the emulsifier component according to polarity such as Tween 20, Tween 80, Span 80, Span 85, Triton X-100.

<Example 12> Confirming the insecticidal effect on the sea layer of the pest control formulation using the active spores of the genus Pacilomis

Insecticidal effects of the compositions prepared in <Example 9> to <Example 11> Confirmed.

Specifically, Figure 5a to 5 (1) harvested strawberry leaves infected with aphids at the strawberry farm site using the facility house as shown in 1 and accumulate 102 aphid larvae per each leaf surface in the laboratory, lx 10 for each test formulation ⁶ to ΙχΙΟ ⁸ concentrations of activated spores were adjusted to 10-fold and sprayed for 6 days.

As a result, as shown in Table 1, the active spores of the purified Pacillomyces lysinus HY-4 strain, which did not complete the formulation, did not achieve the insecticidal effect as an indicator of efficacy, but stabilized such as hydrating agents and emulsions. The formulation was confirmed to show a pesticidal effect of 50-100% (number of dead aphids / number of living aphids). In particular, emulsions containing active spores at concentrations of Ι ^χ ΙΟ ⁷ and Ι ^χ ΙΟ ⁸ after 6 days of spraying were found to exhibit 100% fleshing effect. Therefore, it was confirmed that the formulation using active spores of the Pacillomyces lysinus HY-4 strain improves the stability of the active spores (Table 1).

Table 1

Insecticidal activity against silkworm larvae

Industrial Applicability

As described above, the present invention relates to a method for preparing a friendly organic organic material using a strain of genus Pasylomyces 0¾ec // o7 or ces), or a biopesticide manufacturing method, and the active spores produced from the strains of genus Pacylomyces. The pesticide-controlled wetting emulsion or oil composition prepared by mixing with an excipient, an emulsifier and the like under optimum conditions is easily dispersed, suspended or mixed in water or an organic solvent, maintains its own marine insect repellent effect, and increases stability. It can be usefully used for controlling sea layers.

Claims

[Range of request]

[Claim 1]

5 to 10 parts by weight of the culture solution of the strain of genus aec // o / Byces or its active spores, 30 to 50 parts by weight of corn starch or potato starch, and 30 to 50 parts by weight of the excipient composition And 30 to 50 parts by weight of dextrin.

[Claim 2]

The insecticide composition for controlling pests according to claim 1, wherein the genus Pacylomyces strain is a Pacylomyces lysinus HY-4 strain deposited with accession number KCTC 0395BP.

[Claim 3]

According to claim 1, wherein the excipient is bentonite, kaolin or zeolite hydrating composition for pest control, characterized in that.

【Claim ⁴ 】

According to claim 1, wherein the composition is a pest control hydration composition, characterized in that the pest control activity and stability is improved.

[Claim 5]

Pest control, including 5 to 10 parts by weight of the culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, and 2 to 5 parts by weight of an emulsifier, based on the total weight of the emulsion composition. Emulsion composition for use.

[Claim 6]

The method of claim 5, wherein the vegetable oil is any selected from the group consisting of corn oil, soybean oil, grape seed oil, canola oil, olive oil, kerosene and animal oil Emulsion composition for controlling pests, characterized in that at least one.

[Claim 7]

According to claim 5, wherein the emulsifier Tween 20, Tween 80, Span 80, Span 85 and Triton X-100, pest control emulsion composition for controlling pests, characterized in that at least one selected from the group consisting of.

[Claim 8]

Oil composition A pesticide control oil composition comprising 5 to 10 parts by weight of culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, and 2 to 5 parts by weight of an emulsifier, based on the total weight of the oil composition.

[Claim 9]

The method of claim 8, wherein the vegetable oil is any one or more selected from the group consisting of corn oil, soybean oil, grape seed oil, canola oil, olive oil, kerosene and animal oil composition for controlling pests.

[Claim 10]

According to claim 8, wherein the emulsifier Tween 20, Tween 80, Span 80, Span 85 and Triton X-100 is an oil composition for controlling pests, characterized in that at least one selected from the group consisting of.

[Claim 11]

2) 5 to 10 parts by weight of the culture solution or activated spores of the genus of Pacillomyces obtained in the step 1), 30 to 50 parts by weight of corn starch or potato starch, and 30 to 50 parts by weight of the excipient composition Mixed wealth Method for producing a water-absorbing agent composition for controlling pests comprising the step

[Claim 12]

2) To the total weight of the emulsion composition, 5 to 10 parts by weight of the culture medium or activated spores of the genus of Pacillomyces obtained in step 1), 50 to 60 parts by weight of vegetable oil 30 to 40 parts by weight of water, emulsifier 2 to A method for producing an emulsion composition for controlling sea layer comprising the step of mixing 5 parts by weight.

[Claim 13]

One) . Culturing the strain of genus Pacylomyces to obtain a culture solution or active spores of the genus Pacylomyces strain;

2) 5 to 10 parts by weight of the culture medium or the active spores thereof, 90 to 98 parts by weight of vegetable oil, and 2 to 5 parts by weight of an emulsifier, based on the total weight of the oil composition, the culture solution of the genus of Pacillomyces obtained in step 1) Method for producing an oil composition for controlling pests comprising the step of mixing.

[Claim 14]

5 to 10 parts by weight of the culture medium or active spores thereof, 30 to 50 parts by weight of corn starch or potato starch, and 30 to 50 parts by weight of excipient, based on the total weight of the hydrating agent composition And 30 to 50 parts by weight of dextrin for use as a wetting agent for controlling pests.

[Claim 15]

A composition comprising 5 to 10 parts by weight of a culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, and 2 to 5 parts by weight of an emulsifier, based on the total weight of the emulsion composition. As emulsion for controlling sea layer Intended use.

[Claim 16]

Using a composition containing 5 to 10 parts by weight of culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, and 2 to 5 parts by weight of an emulsifier, based on the total weight of the oil composition, as an oil for controlling pests Use for.

[Claim 17]

5 to 10 parts by weight of the culture medium or active spores thereof, 30 to 50 parts by weight of corn starch or potato starch, 30 to 50 parts by weight of excipient, and 30 to 50 parts by weight, based on the total weight of the hydrating agent composition. A pest control method comprising the step of treating a plant, soil or seed with a hydration composition for pest control comprising a weight part.

[Claim 18]

For pest control comprising 5 to 10 parts by weight of culture medium or active spores thereof, 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, and emulsifier 2 to 5 parts by weight, based on the total weight of the emulsion composition. A method for controlling sea layer comprising treating an emulsion composition to a plant, soil or seed.

[Claim 19]

To the pesticide control oil composition comprising 5 to 10 parts by weight of the culture medium or active spores thereof, 90 to 98 parts by weight of vegetable oil, and 2 to 5 parts by weight of the emulsifier, based on the total weight of the oil composition. , Pest control method comprising the step of treating the soil or seeds.

[Claim 20]

The composition of claim 17, wherein the composition is a liquid. Immune to plants, immersed or sprayed on the seeds of the crop in the state or pest control method characterized in that the coating to the seed used.