KR101661566B1 - Manufacturing method of Biopesticide using Paecilomyces sp - Google Patents

Abstract

The present invention in that, the optimum conditions for the active spore production specifically Indeed the L from my process in the strain as an excipient, an emulsifying agent relating to affinity light organic material, or a biological pesticide manufacturing method using the my process (Paecilomyces) sp chamber L The emulsifier, emulsion or oil composition for pest control prepared by mixing easily disperses, suspends or mixes in water or an organic solvent, maintains a unique pest insecticide effect and increases stability, and thus can be usefully used for pest control .

Description

{Manufacturing method of Biopesticide using Paecilomyces sp.

The present invention relates to a hand thread debugging method for biopesticide with an activation of the spores My process (Paecilomyces) sp.

Biopesticide is also known as a biocontrol agent or a green pesticide and can be used to treat pathogens, pests and other organisms that damage the production and preservation of crops using substances in the natural environment and materials derived therefrom. Weeds and other crops. The US EPA classifies biochemical pesticides derived from natural products, microbial pesticides using microorganisms controlling insects, and plant-incorporated protectants (PIPs). In the EU, it refers to pesticides using microorganisms or natural products. In Korea, the concept of the same or similar to biological pesticides is to protect the crop protection function component inherently possessed by natural biological resources (plants, microorganisms, insects, etc.) from the natural or harmful substances or products produced for the purpose of crop pest control without the artificial chemical treatment process. It is classified as crop protection material.

Crop protective materials are classified as biological pesticides and chemical pesticides, and biological pesticides are classified as biological pesticides including microbial pesticides, biochemical pesticides, and others using natural enemies, nematodes, and organisms. Among these pesticides, microorganisms and natural materials are used to classify organic farming materials as organic materials.

For centuries, agriculture has developed with the aim of mass production and consumption, and domestic and overseas agriculture and industry in the 21st century emphasized the safety of food as well as the need for stable food and well-being according to the abnormal climate. . Therefore, the demand for organic agricultural products is increasing rapidly, and the cultivation area is continuously increasing. In particular, the issue of safe food is related to the primary safety of the crops produced. Therefore, the government is establishing a plan to foster environmentally-friendly agriculture, expanding the production proportion of environmentally friendly agricultural products, and continuously reducing the use of chemical pesticides and fertilizers .

In order to secure the international competitiveness of agriculture and industry in Korea, the production of environmentally safe agricultural products has become inevitable, and the development and dissemination of excellent biocidal pesticides that can replace chemical pesticides is essential. This is an indispensable factor for the government to increase the proportion of production of environment-friendly agricultural products by 10% and to reduce the amount of chemical pesticides by 40% by 2013 in the 5-year plan for the second eco-

Currently, there are more than 370 registered biologically active pesticides in the world, of which about 270% are pesticides, and the development of bio-pesticides is mainly carried out by small companies. The development of microbial pesticides based on microorganisms is a major feature. Microbial pesticides are the most commonly used microbes registered as fungi and bacteria, followed by pesticides.

Insect Pests Pest control using fungi is an environmentally friendly means of controlling, selectively acting only on target insects, inhibiting the appearance of resistance to pests, allowing continuous control, and no toxicity to the environment, It is possible to produce insect pathogenic fungi suitable for the domestic environment in a wide range of countries. Therefore, it is a practical practice to isolate, mass-produce and formulate fungi suitable for the domestic environment.

In Korea, research and development to date have been limited to the development of insect pathogenic fungi probiotics, making it difficult to achieve industrial achievement. In order to apply the probiotics to the agricultural field, the stability of the formulation should be ensured through the formulation to maintain its activity. However, it is difficult to secure the economical efficiency of the actual developed products and the excellent effects in the field due to the lack of technology related to the related parts. These problems are an important factor in the sluggish market expansion.

The present inventors have wettable powder by mixing the efforts result, L Indeed excipients the active spores produced from my process in the strain, the optimal condition as the emulsifier to develop the biopesticide stability is ensured pest repellent effect and material through the formulation, Emulsion, oil or emulsion is easily dispersed, suspended or mixed in water or an organic solvent, the inherent pest insecticidal effect is maintained, and the stability is improved. As a result, the composition Can be usefully used for pest control, thereby completing the present invention.

An object of the present invention to provide a biological activity for controlling pests using the spores of my process (Paecilomyces) strain indeed L pesticides and their preparation.

In order to achieve the above object,

The present invention is a wettable powder composition total weight losses indeed My three's (Paecilomyces) sp culture medium or its activated spores of 5 to 10 parts by weight with respect to, corn starch or potato starch 30 to 50 parts by weight of excipient 30-50 parts by weight of dextrin And 30 to 50 parts by weight of the composition.

In addition, the present invention emulsions compositions culture or its activity spores L indeed My process in strain with respect to the total weight of 5 to 10 parts by weight, 50 to 60 parts by weight of vegetable oil unit, 30 to 40 parts by weight of water, an emulsifying agent from 2 to 5 parts by weight The present invention provides a pest controlling emulsion composition containing the same.

In addition, the present invention oil for controlling pests comprising the oil agent composition of the culture or its activated spores of my process in strain indeed L, based on the total weight of 5 to 10 parts by weight of a vegetable oil 90 to 98 parts by weight of an emulsifier from 2 to 5 parts by weight / RTI >

In addition,

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of the culture broth or active spore of the Rosaceae Iscense strain obtained in the step 1), 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 of dextrin are mixed with each other.

In addition,

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of a culture solution or active spore of the plant belonging to the genus Lactobacillus strain obtained in the step 1), 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifier 2 To 5 parts by weight of the emulsion-controlling emulsion composition.

In addition,

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of the culture solution of the Rosaceae Iscense strain obtained in the above step 1) or its active spore, 90 to 98 parts by weight of the vegetable oil and 2 to 5 parts by weight of the emulsifier with respect to the total weight of the oil composition And an oil-in-water emulsion composition.

The present invention relates to L indeed My process (Paecilomyces) prepared by mixing at the best condition as an excipient, an emulsifying agent for the activated spores produced from sp wettable powder for controlling pests, emulsions or oil compositions, the composition is water or an organic solvent And the insect pest insecticide effect is maintained, and the stability is increased, so that it can be usefully used for pest control.

Figure 1 is a confirm the stability of the excipient for the dispersion, a suspension or a mixture of activated spores of chamber L my process will Sinners (Paecilomyces lilacinus) HY-4 strain.
2 is a diagram showing a storage period (days) spore viability (%) of activated spores of Sinners HY-4 Strain L will indeed My process, including a variety of emulsions.
Figure 3 is a process chamber will my hand for checking the stability of the emulsions of the active spore Sinners HY-4 strain.
Figure 4 is a check to improve stability of the active spores Sinners HY-4 Strain L will indeed My process for the antioxidant.
Figure 5 is a confirm the insecticidal effect of the wettable powders and emulsions, which will My process chamber L containing active spores Sinners HY-4 strain.

Hereinafter, the present invention will be described in detail.

The present invention is a wettable powder composition total weight losses indeed My three's (Paecilomyces) sp culture medium or its activated spores of 5 to 10 parts by weight with respect to, corn starch or potato starch 30 to 50 parts by weight of excipient 30 to 50 parts by weight, and And 30 to 50 parts by weight of dextrin.

Indeed, the hand my process in strain will My process chamber to deposit the L accession No. KCTC 0395BP Sinners (Paecilomyces lilacinus) which is one preferred HY-4 strain, but not limited thereto.

It is preferred that the excipient is free of toxicity to microorganisms and that it is acceptable to use as agricultural soil treatment agent. It is more preferable that the sprayer is not clogged with a nozzle and can control the moisture content, and most preferably bentonite, kaolin or zeolite But is not limited thereto.

The above-described wettable powder for insect controlling composition preferably has improved pest control activity and stability, but is not limited thereto.

The above-described wettable powder composition may further include known auxiliary agents as needed. The adjuvants may include, but are not limited to, stabilizers, antioxidants, thickeners, defoamers, and the like.

Culture broth or its activated spores of the L indeed My process (Paecilomyces) in strains of wettable powder compositions, if containing less than 5 parts by weight based on total weight, since the pest control effect significantly reduced, indeed the L culture of my process spp or The active spore thereof is preferably contained in an amount of 5 to 10 parts by weight based on the total weight of the composition, but is not limited thereto.

Culture broth or its activated spores of the L indeed My process (Paecilomyces) in strains of wettable powder compositions, if containing at least 10 parts by weight based on the total weight, is contaminated by floating fungus in the air is generated, my process spp Indeed the L of the culture medium or so thereof, a phenomenon that viability is remarkably reduced in the active spore is generated pest control effect is significantly reduced, the L indeed culture of my process in strain or its activated spores wettable powder composition of 5 to 10 with respect to the total weight of parts by weight But is not limited thereto.

When the corn starch or potato starch contains not more than 30 parts by weight or not less than 50 parts by weight based on the total weight of the wettable powder composition, the wettable powder composition is not significantly mixed or suspended in water, The corn starch or potato starch is preferably contained in an amount of 30 to 50 parts by weight based on the total weight.

The above-described wettable powder composition should be designed to have appropriate powderiness, dispersibility and moisture content so as to be in a state of suspension when mixed with water. When the wettable powder composition contains not more than 30 parts by weight or not less than 50 parts by weight of the excipient, The germination rate and the suspension property of the spore are remarkably decreased and the pest control activity is decreased, so that the excipient is preferably contained in an amount of 30 to 50 parts by weight based on the total weight.

In a particular embodiment of the present invention, the inventors have deposited with the after separating a microorganism having an insecticidal effect on pests from the soil, identified and will My process chamber L named Sinners HY-4, and Korea Research Institute of Bioscience and Biotechnology Gene Bank (Accession No. KCTC 0395BP).

In addition, the present inventors have then to ensure the stability of the vehicle for dispersing, suspending and mixing of my process spp active spore indeed L, mixing the L indeed My process spp activated spores and excipients, storage period spore germination number of viable cells , It was confirmed that the germination rate of active spores was maintained at 70% or more even when the active spore mixed with bentonite, kaolin or zeolite was stored for 6 months or more (see FIG. 1).

In addition, the present inventors on, then a solution of the excipients for the active spores, a result of measuring the respective concentrations by suspension of the castle, wettable powder composition total weight to chamber L by the vehicle to determine the suspension of the My process spp activated spores Was contained in an amount of 30 parts by weight to 50 parts by weight based on 100 parts by weight of the composition.

In addition, the present inventors have wettable powder composition total weight losses indeed My three's (Paecilomyces) sp culture medium or from 5 to 10 parts by weight, its activity spores with respect to, corn starch or potato starch 30 to 50 parts by weight, and excipients from 30 to 50 wt. , And the insecticidal effect was confirmed. As a result, it was confirmed that the purified active spore did not exhibit a significant insecticidal effect, while the wetter composition of the present invention showed an insecticidal effect of 50 to 100% 5 and Table 1).

Thus, since the wettable powder composition of the invention is to facilitate the dispersion and mixing and suspending the active spore production chamber from my process spp L in water, and to maintain and stabilize the unique pest control activity, useful in pest control Can be used.

In addition, the present invention L Indeed MY culture medium of the process in strain or its activated spores of 5 to 10 with respect to the total weight of the emulsifier composition parts by weight, 50 to 60 parts by weight of vegetable oil unit, 30 to 40 parts by weight of water, an emulsifying agent from 2 to 5 parts by weight The present invention provides a pest controlling emulsion composition containing the same.

Indeed, the hand my process in strain will My process chamber to deposit the L accession No. KCTC 0395BP Sinners (Paecilomyces lilacinus) which is one preferred HY-4 strain, but 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 emulsion controlling emulsion composition preferably has improved pest control activity and stability but is not limited thereto.

The emulsion composition may further include known auxiliaries, if necessary. The adjuvants may include, but are not limited to, stabilizers, antioxidants, thickeners, defoamers, and the like. The antioxidant may be selected from the group consisting of tertiary-butylhydroquinone (TBHQ), butylhydroxyl toluene (BHT), butylated hydroxyl anisole (BHA), propyl gallate But is not limited to, vitamin C (vitamin C, Vt.C), vitamin E (vitamin E, Vt.E), lecithin or iron oxide (Fe ₂ O ₃ ).

Culture broth or its activated spores of the L indeed My process (Paecilomyces) in strains emulsions composition, if containing less than 5 parts by weight based on total weight, since the pest control effect significantly reduced, indeed the L culture of my process spp or The active spore thereof is preferably 5 to 10 parts by weight based on the total weight of the emulsion composition, but is not limited thereto.

The L indeed My process (Paecilomyces) culture medium or active spores thereof in strain emulsions composition, if containing at least 10 parts by weight based on the total weight, is contaminated by floating fungus in the air is generated, my process spp Indeed the L of the culture medium or so thereof, a phenomenon that viability is remarkably reduced in the active spore is generated pest control effect is greatly reduced, indeed the L culture of my process in strain or its activated spores is 5 to 10 with respect to the total weight of the emulsions compositions parts by weight But is not limited thereto.

The emulsifier preferably contains 2 to 5 parts by weight based on the total weight of the emulsion composition, and when the emulsion composition contains 2 parts by weight or less, the emulsion composition of the present invention is not mixed or suspended in a solvent.

In a specific example of the present invention, in order to examine the stability of active spores with respect to various tanning agents, the present inventors suspended active spores of the present invention in corn oil, soybean oil, etc., and then confirmed the germination of spores and measured the survival rate As a result, it was confirmed that the tanning agent such as corn oil had no effect on germination of spores (see Fig. 2).

Further, in order to examine the stability of active spores with respect to various emulsifiers, the inventors of the present invention measured the survival rate by confirming the germination of spores after adding active spores of the present invention to soybean oil, adding an emulsifier, It was confirmed that the emulsifier did not significantly affect germination of spores until 6 months (see FIG. 2).

In addition, the inventors of the present invention confirmed that the active spores added with an antioxidant maintained the spore survival rate of 88% or more even after 210 days (see FIG. 4) .

Accordingly, the emulsions the compositions of the present invention is the active spores produced from my process spp chamber L to facilitate the dispersion, mixing and suspended in water and the solvent, it is possible to maintain and stabilize the unique pest control active, the debugging Can be usefully used.

In addition, the present invention oil for controlling pests comprising the oil agent composition of the culture or its activated spores of my process in strain indeed L, based on the total weight of 5 to 10 parts by weight of a vegetable oil 90 to 98 parts by weight of an emulsifier from 2 to 5 parts by weight / RTI >

Indeed, the hand my process in strain will My process chamber to deposit the L accession No. KCTC 0395BP Sinners (Paecilomyces lilacinus) which is one preferred HY-4 strain, but 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 above-mentioned oil composition for controlling pests is preferably, but not limited to, improved pest control activity and stability.

The oil composition may further include known auxiliaries, if necessary. The adjuvants may include, but are not limited to, stabilizers, antioxidants, thickeners, defoamers, and the like. The antioxidant may be selected from the group consisting of tertiary-butylhydroquinone (TBHQ), butylhydroxyl toluene (BHT), butylated hydroxyl anisole (BHA), propyl gallate But is not limited to, vitamin C (vitamin C, Vt.C), vitamin E (vitamin E, Vt.E), lecithin or iron oxide (Fe ₂ O ₃ ).

When the culture or its activated spores of the L indeed My process (Paecilomyces) in strains containing 5 parts by weight or less with respect to the oil agent composition total weight, since the pest control effect is significantly reduced, the culture medium of my process spp Indeed the L Or active spore thereof is preferably 5 to 10 parts by weight based on the total weight of the emulsion composition, but is not limited thereto.

The L indeed My process (Paecilomyces) culture medium or active spores thereof in strain emulsions composition, if containing at least 10 parts by weight based on the total weight, is contaminated by floating fungus in the air is generated, my process spp Indeed the L of the culture medium or so thereof, a phenomenon that viability is remarkably reduced in the active spore is generated pest control effect is greatly reduced, indeed the L culture of my process in strain or its activated spores is 5 to 10 with respect to the total weight of the emulsions compositions parts by weight But is not limited thereto.

The emulsifier preferably contains 2 to 5 parts by weight based on the total weight of the oil composition, and when the oil composition contains 2 parts by weight or less, the oil composition of the present invention is not mixed or suspended in a solvent, .

Thus, the oil agent composition of the present invention and the active spore production chamber from my process spp L to facilitate the dispersion, mixing and suspension in a solvent, it is possible to maintain and stabilize the unique pest control activity, useful in pest control Lt; / RTI >

The present invention also provides a process for the preparation of pest control wettable powder, emulsions or oil agent composition using a liquid culture or spores of active My process sp chamber tiles.

The above-described wettable powder composition is preferably prepared by the following method, but is not limited thereto:

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of the culture broth or active spore of the Rosaceae Iscense strain obtained in the step 1), 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 of dextrin.

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

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of a culture solution or active spore of the plant belonging to the genus Lactobacillus strain obtained in the step 1), 50 to 60 parts by weight of vegetable oil, 30 to 40 parts by weight of water, emulsifier 2 ≪ / RTI > to 5 parts by weight.

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

1), to chamber L culture process in the MY strain comprising: L indeed give a culture or spores of active My process spp;

2) 5 to 10 parts by weight of the culture solution of the Rosaceae Iscense strain obtained in the above step 1) or its active spore, 90 to 98 parts by weight of the vegetable oil and 2 to 5 parts by weight of the emulsifier with respect to the total weight of the oil composition .

Indeed, the hand my process in strain will My process chamber to deposit the L accession No. KCTC 0395BP Sinners (Paecilomyces lilacinus) which is one preferred HY-4 strain, but not limited thereto.

The composition prepared in the step 2) may further include known auxiliary agents as necessary. The adjuvants may include, but are not limited to, stabilizers, antioxidants, thickeners, defoamers, and the like.

Therefore, the present the wettable powder, emulsions or oil agent composition prepared by the method of the invention is to facilitate the dispersion and mixing and suspending the active spore production chamber from my process spp L in water or in a solvent, a unique pest control active And can be used for pest control.

Hereinafter, the present invention will be described in detail with reference to examples.

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

≪ Example 1 > Facilomyces Lira sinus  Preculture of strain HY-4

A PDA (Potato Dextrose Agar) medium was used for cultivation of the seedlings for storage and cultivation of the Fasylomyces lysineus HY-4 strain (Accession No. KCTC 0395BP; Korean Registration Number: 10-0475131). The seed culture was cultured in petri dish (87 × 15 mm) at 28 ° C. and 65% humidity in a PDA medium for 7 days in a dark room.

≪ Example 2 > Facilomyces Lira sinus  Culture of strain HY-4

For the cultivation of Fasylomyces lysinus HY-4 strain, the mycelia were maximized in the culture medium by liquid culture in the primary culture, and then solid culture, which is a secondary culture, was carried out for preparing insecticides.

Specifically, for the mycelial culture, the first liquid culture was cultured in PBD medium at 28 DEG C and 180 rpm for 7 days. Then, the cells were cultured in a Petri dish (150 × 25 mm) under the condition of 25 ml of Czapek medium, and 2 ml of the mycelium-containing primary cultures were dispensed and spread in a spreader for solid culture for 20 days.

≪ Example 3 > Facilomyces Lira sinus  Spore recovery of strain HY-4

In order to collect spores of the Lycoris sylarisenus HY-4 strain formed through solid culture for 7 days in the Petri dish of Example 2, the hymen was dried for 10 days in a hume hood to completely dry the culture medium. The spores were recovered through a scraper in a culture medium of a dried Petri dish.

Example 4 Confirmation of Stability of Excipients for Dispersion, Suspension and Mixing of Active Spores

The <Example 3> and L will indeed My process recovery was confirmed in the stability of the vehicle by using the activated spores Sinners HY-4 strain.

Specifically, the microorganisms such as bentonite, kaolin, and zeolite, which are mainly used as excipients for biological pesticides or environmentally friendly organic materials, and polysaccharides such as dextrin, corn starch, and rice bran were used to measure the number of spore germ viable cells . The spore viability was measured every 30 days while 250 ml brown bottle (Amber bottle, SPL) containing 100 g mixture was mixed at a ratio of spore to excipient of 1: 9 (w: w) and kept at room temperature. The survival rate of the spores was determined by suspending 0.5 g of the sample in 4.5 ml of physiological saline, stepwise diluting in 1/10 of each, diluting 0.1 ml of the diluted solution uniformly onto the PDA plate medium, culturing at 28 ° C for 2 days And then colonies were measured.

As a result, as shown in Fig. 1, it was confirmed that the spore germination rate varied depending on each excipient from the storage period of 4 months. Especially, the spore germination rate of specimens containing no excipients after 6 months decreased to 50% or less, while the spores containing excipients were maintained for more than 50% even after storage for 11 months One).

Example 5 Confirmation of Suspension of Active Spores Using Excipients

The excipients for the <Example 3> and L will indeed recovered in my process the suspension of the activated spores Sinners HY-4 strain was confirmed by wetting time (wetting time).

Concretely, a suspension of various minerals such as bentonite, kaolin and zeolite and polysaccharides such as dextrin, corn starch and rice bran were mixed and measured for each concentration. The ratio of spore to excipient was 1: 9 (w: w) to 10: 0 (w: w), and 1 g of composition was mixed with a conical tube (SPL) (W1). After standing at room temperature for 30 minutes, 40 ml of the supernatant was precisely removed with a pipette, and the remaining suspension was dried to measure the dry weight (W2).

Suspension (%, w / w) was calculated by the following equation.

As a result, a composition comprising 10% (w / w) spore and 90% (w / w) mineral has a solubility of 85-92% (w / w) (w / w) polysaccharide in a composition comprising 25-35% (w / w) suspension, 10% (w / w) spore, 45% polysaccharide, 45% To 75% (w / w).

Example 6 Confirmation of Stability of Active Spores on Various Emulsions

The stability of the emulsion the various <Example 3> and L will indeed recovered in my process using activated spores Sinners HY-4 strain was confirmed.

A specific, corn oil, soybean oil, grape seed oil, canola oil, various kinds of vegetable oils suspended activated spores 1% (w / v) of L will indeed My process Sinners HY-4 strains of the olive oil, etc. and stored at room temperature following. The survival rate of each storage sample was determined by confirming the germination of spores every 30 days for 180 days. The survival rate of the spores was determined by suspending 0.5 g of the sample in 4.5 ml of physiological saline, stepwise diluting in 1/10 of each, diluting 0.1 ml of the diluted solution uniformly onto the PDA plate medium, culturing at 28 ° C for 2 days And then colonies were measured.

As a result, as shown in FIG. 2, the survival rate of spores was remarkably decreased in the case of the oil-free emulsion starting from 120 days after storage, and the survival rate was maintained at 50% after 180 days. On the other hand, Maintained a spore survival rate of 90% or more (Fig. 2).

Example 7 Confirmation of Stability of Active Spores for Various Emulsifiers

The stability of the various emulsifying the <Example 3> and L will indeed recovered in my process using activated spores Sinners HY-4 strain was confirmed.

Specifically, the soybean oil will indeed lost my process Sinners HY-4 activated spores 5% (w / v) Tween 20 and the strain, Tween 80, Span 80, Span 85 and Triton X-100 (Sigma-Aldrich Co., LLC.) Each emulsifier was suspended in 2.5% (w / v) and stored at room temperature. The survival rate of each storage sample was determined by confirming the germination of spores every 30 days for 240 days. The survival rate of the spores was determined by suspending 0.5 g of the sample in 4.5 ml of physiological saline, stepwise diluting in 1/10 of each, diluting 0.1 ml of the diluted solution uniformly onto the PDA plate medium, culturing at 28 ° C for 2 days And then colonies were measured.

As a result, as shown in Fig. 3, it was confirmed that active spores containing various emulsifiers did not show significant difference in spore survival rate as compared with the control group without emulsifier (Fig. 3).

Example 8 Confirmation of Improvement of Stability of Active Spores for Various Antioxidants Using Antioxidants

The coating suspension or in vegetable oils the <Example 3> A paesil Rome rira process recovery was found in the improved stability of the various antioxidants with an activation of the spores Sinners HY-4 strain.

Specifically, will indeed lost the soybean my process was suspended activated spores 1% (w / v) of Sinners HY-4 strain tertiary butyl hydroquinone, (tertiary-butylhydroquinone, TBHQ), butyl hydroxy toluene (butyl hydroxyl toluene, BHT), butylated hydroxyl anisole (BHA), propyl gallate (PG), vitamin C (vitamin C, Vt.C), vitamin E (vitamin E, Vt.E), lecithin ) And 0.02% (w / v) antioxidant such as iron oxide (Fe ₂ O ₃ ) were added and stored at room temperature. Each storage sample was checked for survival rates at intervals of 30 days for 210 days.

As a result, as shown in FIG. 4, the survival rate of spores stored for 180 days was reduced to 50%, whereas the storage samples containing antioxidants except vitamin C and vitamin E maintained spore survival rate of 80% or more. In particular, after 210 days, it was confirmed that the storage sample to which butylhydroxytoluene was added in each of the storage samples maintained the 88% spore survival rate and the spore stability was the highest (FIG. 4).

&Lt; Example 9 > Facilomyces  Preparation of wettable powders for pest control using activated spores

L will indeed My process to prepare a wettable powder using a mixture or suspended in water for the purpose of a stable formulation of activated spores of Sinners HY-4 strain.

Specifically, the <Example 4> and <Example 5> and as L indeed My recessed ra Sinners HY-4 strain of the active spores from 5 to 10% (w / v), corn starch and 5 to 10% (w / v), 30 to 50% (w / v) of bentonite, and 30 to 50% (w / v) of dextrin were mixed to prepare a wettable powder composition. The bentonite can be substituted with minerals such as kaolin, zeolite and the like. Corn starch can also be substituted for potato starch.

&Lt; Example 10 > Facilomyces  Production of Emulsion Control Pest Control Agent Using Spores

For the purpose of stably formulating the active spore of the Fabilomyces lysinus HY-4 strain, a coagulant was prepared by mixing or suspending the active spore in a solvent and water.

Specifically, the <Example 4> and <Example 6> and as L indeed My recessed ra Sinners HY-4 strain of the active spores from 5 to 10% (w / v), 50 to 60% of vegetable oils (w / v), 30 to 40% (w / v) water, and 2 to 5% (w / v) emulsifier were mixed to prepare a emulsion composition. The vegetable oil may be substituted for kerosene or animal oil widely used for agricultural purposes. The emulsifier may be different from the emulsifier according to polarity such as Tween 20, Tween 80, Span 80, Span 85, and Triton X-100.

&Lt; Example 11 > Facilomyces  Manufacture of an oil agent for pest control using active spores

L yarn was produced my process will Sinners oils using a mixture or a stable suspension of the active formulated spores of HY-4 strains in a solvent for the purpose.

Specifically, the <Example 4> and <Example 6> and as L indeed My recessed ra Sinners HY-4 activated spores from 5 to 10% (w / v) of a strain with a vegetable oil 90 to 98% (w / v) and 2 to 5% (w / v) of an emulsifier were used. The vegetable oil may be substituted for kerosene or animal oil widely used for agricultural purposes. The emulsifier may be different from the emulsifier according to polarity such as Tween 20, Tween 80, Span 80, Span 85, and Triton X-100.

&Lt; Example 12 > Facilomyces  Identification of insecticidal effect of insect control formulation against pests

The insecticidal effect of the compositions prepared in Examples 9 to 11 was confirmed.

Specifically, as shown in FIGS. 5A to 5D, strawberry leaves infected with aphids were collected at a strawberry farm site using a facility house, and then a total of 102 aphid larvae were collected per each strawberry leaf surface in a laboratory, and 1 × 10 ⁶ The concentration of active sporophyll was adjusted to 10 × and concentration was 1 × 10 ⁸ , and the insecticidal effect was analyzed for 6 days.

As a result, will My process chamber purified L did not complete the formulation as shown in Table 1, but the active spores Sinners HY-4 strains has not achieved the insecticidal effect which is an index of the efficacy, the stabilized, such as wettable powder, emulsions The formulation showed insecticidal effects of 50-100% (dead aphids / live aphids). Particularly, it was confirmed that the emulsion containing active spores at concentrations of 1 × 10 ⁷ and 1 × 10 ⁸ after 6 days of spraying showed a 100% insecticidal effect. Thus, my process will indeed L formulated with an activation of the spores Sinners HY-4 strain was confirmed to enhance the stability of the activated spores (Table 1).

Claims (3)

With respect to the emulsions the total composition weight of L will indeed My process deposited as accession number of KCTC 0395BP Sinners (Paecilomyces lilacinus) HY-4 group consisting of activated spores of 5 to 10 parts by weight of corn oil, soybean oil, grape seed oil, canola oil, olive oil of the strain From 30 to 40 parts by weight of water and from 2 to 5 parts by weight of any one selected from the group consisting of Tween 20, Tween 80, Span 80, Span 85 and Triton X-100. &Lt; / RTI &gt;
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