KR101666968B1 - Beauveria bassiana having insect pathogenicityand agent for prevention of rice vermin using the same - Google Patents

Abstract

The present invention relates to an insect pathogenic fungus, bovaria barbiana JEF007 strain, and an agricultural pest using the same, particularly a liquid preparation for horticultural or fruit pest control. The agricultural pest control agent according to the present invention can be easily handled, harmless to the human body, and can be used in an environmentally friendly manner to control agricultural pests by producing the agricultural pest control agent using Beauveria bassiana JEF007 strain, which is an insect pathogenic fungus, It is possible to produce a liquid preparation of the strain simply and economically.

Description

FIELD OF THE INVENTION [0001] The present invention relates to an insect pathogenic bovaria barbiana and a liquid preparation for controlling agricultural pests using the same,

The present invention relates to an insect pathogenic fungus, bovaria barbiana JEF007 strain, and an agricultural pest using the same, particularly a liquid preparation for horticultural or fruit pest control.

It is widely distributed in Korea, Japan, and Taiwan, and has been widely distributed in Korea, such as soybean, soybean, and pea, such as soybean crops, watermelon, melon, cucumber, Is a major agricultural pest causing great damage to the production of crops, which directly sucks rice and crops such as rice, blood, and marsh and suffers physical damage. In order to control pests harmful to these crops, the use of chemical pesticides has been increased and the spreading area has become wider as the size of cultivated crops has been increased with the development of agricultural technology. The emergence of organic synthetic insecticides after World War II marked a milestone turning point in two aspects: increasing agricultural crops by agricultural pest control and improving human health by sanitary pest control. Insecticides used for pest control in Korea are increasing in usage every year, of which organic phosphorus and carbamate accounts for about 90% of total usage.

However, due to decades of continuous use and abuse of these organic synthetic pesticides, various adverse effects such as the appearance of resistant pests, lethal toxicity, biological control disturbance by natural enemies, residual poison and environmental pollution have been caused. Recently, The use of synthetic insecticides is becoming increasingly restricted as the interest in the adverse effects and environmental effects of the insecticides is increasing. Biological control has been actively studied as a next-generation pesticide that can replace synthetic pesticides, but biological control has become a hindrance to commercialization because it is difficult to handle using microorganisms, natural enemies or natural extracts. Studies on biological control using insect pathogenic fungi have been actively carried out, but research on mass production and formulation, which is an important part in commercialization, is insufficient. Currently, insect pathogenic fungi are produced by separating the spores produced from the solid culture from the cultures and then spraying them in the form of a liquid preparation and a wetting agent. However, in the process of harvesting spores, high labor costs and production space There is a problem that is becoming a hindrance to commercialization.

Accordingly, there is a desperate need to develop a new agricultural pest control agent that is harmless to the human body, eco-friendly, easy to handle, and easy to manufacture, and which can be used commercially.

KR 10-2012-0140146

The present inventors newly discovered a strain of JEF007 of Beauveria bassiana while searching for an agricultural pest control agent and confirmed that the strain has an insect pathogenicity and excellent spore forming ability and is used for agricultural pest control The present invention has been accomplished by confirming that a liquid preparation of the above strain can be produced simply and economically by using a filtration bag having an excellent controlling effect against agricultural pests.

Accordingly, the present invention provides B. bassiana JEF007 strain, spore, microbial agent and liquid preparation thereof, which is an insect pathogenic fungus for agricultural pest control.

In order to achieve the above object,

The present invention provides a strain of Beauveria bassiana JEF007 (KCCM 11592P) having insect pathogenicity.

In addition, the present invention provides a microorganism preparation for agricultural pest control, comprising a strain of Beauveria bassiana JEF007 or a spore thereof.

The present invention also relates to a method for producing a bacterium, which comprises culturing a bovaria bacillus JEF007 strain in a solid culture medium; And harvesting the culture. The present invention also provides a method for producing a microorganism preparation for agricultural pest control.

The present invention also relates to a method for producing a bacterium, which comprises culturing a strain of Beauveria bassiana JEF007 in a solid culture medium; Encapsulating the culture in a filter bag; And a step of immersing the filter bag in water to elute a strain of Beauveria bassiana JEF007 or spores thereof.

The present invention also provides a liquid preparation for agricultural pest control, which is produced by the above production method.

Hereinafter, the present invention will be described in detail.

The present invention provides a strain of Beauveria bassiana JEF007 (KCCM 11592P) having insect pathogenicity.

The present invention also provides a composition for controlling agricultural pests, comprising a strain of Beauveria bassiana JEF007 or a spore thereof.

In addition, the present invention provides a microorganism preparation for agricultural pest control, comprising a strain of Beauveria bassiana JEF007 or a spore thereof.

The " Beauveria bassiana JEF007 strain" of the present invention is one of the fungi isolated from natural soil by the present inventors and exhibits excellent insecticidal effect against agricultural insect pests, for example, The bacterium Bovia bacillus was isolated by a biochemical assay, and the isolated strain was morphologically identified and analyzed for its genomic DNA sequence to show about 98% homology with the known bobbia bacillus strains It was named as JEF007 and deposited on Oct. 28, 2014 with the Korean Society for Microbiological Protection (KCCM11592P).

The bovaria bacillus JEF007 strain has a morphological characteristic of forming white clusters and spores (see Fig. 1). The JEF007 strain has particularly excellent spore forming ability, and the spore of the JEF007 strain is superior to the known bovaria bacillus or GHA strain for spore storage stability.

The insect pathogenicity refers to the ability of a pathogen to infect a subject insect (host insect) to cause disease. In the present invention, the boveriac basin or JEF007 strain is infected with agricultural pests, It means to cause illness.

The insect pathogenicity may be an insecticidal activity. Specifically, the bovaria bacillus or JEF007 strain or the spore of the strain attaches to the outer shell of the host insect and germinates to produce chitinase, protease, lipase, esterase, etc. And penetrates into the cuticle layer of the insect and propagates in the insect's body, so that a small molecular weight produces a toxic protein such as a toxic substance or a glycoprotein to block the immune function of the insect and kill the host insect.

The bovaria bacillus JEF007 isolate has excellent spore forming ability and has a remarkably excellent insecticidal activity against agricultural pests, and can be effectively used for controlling agricultural pests.

The insect can be a pest, and the pest includes all kinds of sanitary pests, forest pests and agricultural pests.

The insect may preferably be an agricultural pest, and the agricultural pest includes both horticultural insect, fruit insect and rice insect. The agricultural insect pests are preferably selected from the group consisting of sawtooth antlers, flower yellow thrips, cabbage worms, greenhouse powder, cigarette louis, licorice moth, peach moth, apple leafy moth, apple moth leaf moth, White lantern worms, rice weevils, rice bran jellyfishes, rice bran jellyfish, and the like, and more preferably, they may be sawdust waist or flower yellow thrips.

The spore of the bovaria bacillus JEF007 strain can be produced by culturing the inoculum of the bovaria bacillus or JEF007 strain in a solid culture medium. The bovaria bacillus JEF007 strain of the present invention can exhibit spore productivity of about 2 to 3 times as compared with the conventional bovaria bacillus or GHA strain.

The microorganism preparation for controlling agricultural pests may be provided as a granular preparation, a liquid preparation or a compost. The microorganism preparation for agricultural pest control has an excellent insecticidal effect when sprayed on agricultural pests. Especially, when compared to the case of spraying microbial agent of the known strain Bobelia basiana or GHA strain, And showed more excellent insecticidal effect in a short period of time.

The microorganism preparation can be preferably a granular preparation, and can be easily handled when the preparation is a granular preparation, and granular preparations can be easily produced using the spore-forming ability of bovia bacillus or JEF007 strain, The production efficiency of the control agent can be improved.

The present invention also relates to a method for producing a bacterium, which comprises culturing a bovaria bacillus JEF007 strain in a solid culture medium; And harvesting the cultivated plant. The present invention also provides a method for producing a microorganism preparation for agricultural pest control.

Preferably, the solid culture medium comprises any one selected from the group consisting of crude, millet, white rice, wheat bran, millet, and combinations thereof, and more preferably includes a batch.

The method for preparing a microorganism preparation for agricultural pest control may further comprise preparing a solid culture medium by mixing any one selected from organic acids with crude, millet, white rice, wheat bran, sorghum and combinations thereof.

The organic acid is not particularly limited as long as it can be used as a carbon source. More specifically, it is preferably any one selected from citric acid, propionic acid, butyric acid, lactic acid, succinic acid, clonic acid, valeric acid, caproic acid and combinations thereof, more preferably citric acid.

The organic acid and the solid culture medium may be mixed by adding an organic acid to the solid culture medium and mixing the organic acid.

The culturing vessel of the strain may be a culture vessel commonly used in the technical field of the present invention, but is not limited thereto. For example, a plastic container for solid culture or a polyvinyl bag can be used for culturing by attaching an oxygen supply filter.

Preferably, the culture of the bovaria bacillus JEF007 strain is carried out at 20 to 30 DEG C for 4 to 8 days, more preferably at 25 DEG C for 6 days. In the present invention, the solid culture medium mixed with the organic acid was heated and cooled at 110 to 130 ° C for 20 to 40 minutes for sterilization, and the culture was carried out by inoculating the bovaria barbiana strain JEF007 into the medium.

The inoculum to be used for the solid culture medium can be obtained by subculturing the bovaria barbiana strain JEF007, and the cultivation of the JEF007 strain can be carried out according to a conventional subculture method. The medium required for the subculture is not limited to that kind.

As an example of the subculture medium, a liquid medium containing PDB (Potato Dextrose Broth) medium, rice bran or wheat bran can be used, but it is not limited thereto. The subculture medium may also include corn flour, rice bran or cornsteep powder, and the like.

The present invention also relates to a method for producing a bacterium, which comprises culturing a strain of Beauveria bassiana JEF007 in a solid culture medium; Encapsulating the culture in a filter bag; And a step of immersing the filter bag in water to elute a strain of Beauveria bassiana JEF007 or spores thereof.

Preferably, the solid culture medium comprises any one selected from the group consisting of crude, millet, white rice, wheat bran, millet, and combinations thereof, and more preferably includes a batch. The culture can be used as it is enclosed in a bag as it is, but it is preferably pulverized. The widening of the surface area of the culture through the pulverization process widens the surface in contact with water, so that the strain and spores thereof are easily eluted from the filterable bag.

The filter bag is a bag made of a water-permeable filter material. The filter bag may be a synthetic fiber of polyester, nylon, polyethylene, polypropylene and vinylon, a semi-synthetic fiber such as rayon, a woven or nonwoven fabric consisting of natural fibers of mulberry or mulberry tree alone or a composite fiber, Wood pulp, mixed paper made of polypropylene fiber, and paper.

The present invention also provides a liquid preparation for agricultural pest control, which is produced by the above production method.

When the bovaria bacillus JEF007 strain, its spore, microorganism preparation or liquid formulation is sprayed on agricultural pests, it is possible to obtain a superior control rate against the ants of the sawdust ants, compared to the case of spraying with the conventional bovaria bacillus or GHA strain There is an effect indicating.

The agricultural pest control agent according to the present invention can be easily handled, harmless to the human body, and can be used in an environmentally friendly manner to control agricultural pests by producing the agricultural pest control agent using Beauveria bassiana JEF007 strain, which is an insect pathogenic fungus, It is possible to produce a liquid preparation of the strain simply and economically.

Brief Description of the Drawings Fig. 1 is a diagram showing an image obtained by observing a bovaria bacillus JEF007 strain cultured in Sabraud dextrose agar (SDA) medium in a dark room at 25 캜 for 7 days with an optical microscope (400 times).
Fig. 2 is a diagram showing the DNA sequencing results of the bovaria bacillus or JEF007 strain.
3 is a view showing a plastic container for solid culture and a microbial preparation of bovaria barbiana strain JEF007, respectively.
FIG. 4 is a diagram showing a method for producing a diluted spray liquid formulation using bovaria bacillus or JEF007 strain.
FIG. 5 is a view showing the effect of controlling the dilution-spray-type liquid preparation using the bovaria bacillus or JEF007 strain against the sawtooth antler.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

Example 1 Isolation and Identification of Strain

1-1. Isolation of strain

Beauveria bassiana The JEF007 strain is a bacteriocin that exhibits excellent insecticidal effect against horticultural and fruit insects, for example, sawtooth ants, and fungi, among the fungi isolated from natural soil. Respectively.

1-2. Growth and morphological identification

An optical microscope (400X) image of the bovaria bacillus JEF007 strain cultured in Sabraud Dextrose Agar (SDA) for 7 days in a dark room at 25 ° C is shown in FIG.

As shown in Fig. 1, the formation of white mycelia, spores, and spores, typical features of the bovaria strain, was well observed.

1-3. Sequencing

After amplification of PCR (polymerase chain reaction, polymerase chain reaction) by ITS1 forward primer and ITS4 reverse primer, The results of DNA sequencing of the bovaria bacillus JEF007 strain are shown in Fig.

Example 2: Preparation of a microorganism preparation for agricultural pest control

200 g of the Italian millet was placed in a plastic container (1 L) for solid culture, and then 0.32 ml of 50% citric acid was added. Thereafter, it was heated at 121 占 폚 for 30 minutes for sterilization. The sterilized bath was cooled to room temperature, and 1 ml of the boveriacacia JEF007 strain cultured in PDB medium for 3 days at room temperature was inoculated into the flask, followed by shaking for 6 days at 25 ° C. to prepare a microorganism preparation of strain JEF007 .

The plastic container for the solid culture and the microorganism preparation of the bovaria barbiana strain JEF007 are shown in Fig. 3, respectively.

Example 3. Confirmation of spore production ability

1 g of the microorganism preparation of strain JEF007 was diluted with 10 ml of 0.3% silwet, and the number of spores was counted using a hamacytometer to measure the spore productivity. The porosity coefficients of the bovaria bacillus JEF007 strain are shown in Table 1.

Strain name Porosity B. bassiana JEF007 2.11 x 10 < ⁸ > conidia / g B. bassiana GHA 8.23 x 10 ⁷ conidia / g

As shown in Table 1, the bovaria bacillus JEF007 strain showed a spore productivity twice as much as that of the conventional commercialized bovaria bacillus or GHA strain.

Example 4: Preparation of dilute spray liquid formulations

10 g of the microorganism preparation of strain JEF007 was placed in a screen bag (8 × 12 cm ² ) and sealed. Thereafter, the sealed filtration bag was impregnated with 20 L of water and shaken for several seconds, thereby releasing the strain or spore thereof in the filter bag into water to prepare a diluted spray-type liquid preparation.

The production method of the diluted spray liquid formulation using the bovaria bacillus JEF007 strain is shown in Fig.

EXPERIMENTAL EXAMPLE 1 Confirmation of the Effect of Diluent Spraying Liquid Formulation on the Back Leg Anchor

The diluted spray-type liquid formulation prepared according to Example 4 was sprayed on a third instar larvae of thirty legged ant thrice larvae of thrips grown in Kangnam-gun using an atomizer. After 3, 7 and 10 days after the spraying, the raw water was examined. The above procedure was repeated three times, and the effect of controlling the back leg antler was examined in FIG. In addition, B. bassiana GHA strain was prepared in the same manner as JEF007 formulation and then spore suspension was treated.

As shown in FIG. 5, the diluted spray formulations using the bovaria bacillus JEF007 strain showed a superior control effect against the antimicrobial agent of the saw blight than the diluted spray formulations using the conventional microorganism, bovaria bacillus or GHA strain. After 7 days of the suspension treatment, the germination rate of the GHA strain was 20% lower than that of the GSS strain, but the JEF007 treatment showed a high mortality rate of over 65%. Also, similar trend was observed in the 10th day survey.

Korea Microorganism Conservation Center KCCM11592P 20141028

Claims (9)

Culturing the strain of Beauveria bassiana JEF007 (KCCM11592P) having an insect pathogenicity in a solid culture medium to obtain a culture;
Encapsulating the culture in a filter bag; And
And the step of immersing the filter bag in water to elute the strain of Beauveria bassiana JEF007 or spores thereof. The method according to claim 1,
The above insects include insect pests such as sawtooth ants backyard, flower yellow thrips, cabbage worms, greenhouse powdery mildew, tobacco roe, licorice moth, peach moth, apple leafy moth, apple moth leaf moth, Wherein the insect pest is any one of pests selected from the group consisting of weevils, rice paddies, rice paddies, and combinations thereof. delete delete The method according to claim 1,
Wherein the solid culture medium comprises any one selected from the group consisting of crude, millet, white rice, wheat bran, millet, and combinations thereof and an organic acid. The method according to claim 1,
Wherein the cultivation is carried out at 20 to 30 DEG C for 4 to 8 days. delete The method according to claim 1,
The filter bag may be a synthetic fiber of polyester, nylon, polyethylene, polypropylene and vinylon, a semi-synthetic fiber such as rayon, a woven or nonwoven fabric consisting of natural fibers of mulberry or mulberry tree alone or a composite fiber, Wherein the paper is formed by any one selected from the group consisting of wood pulp, mixed paper made of polypropylene fiber, and paper. delete

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