KR102586228B1 - A composition comprising Pediococcus sp. to control Protaetia brevitarsis fungal diseases caused by Metarhizium anisopliae - Google Patents

Abstract

The present invention relates to a composition that can treat or prevent fungal diseases of white-spotted flower radish, preferably green-gold disease, containing microorganisms of the genus Pediococcus, and a feed composition that can promote the growth of white-spotted flower radish.

Description

A composition comprising Pediococcus sp. to control Protaetia brevitarsis fungal diseases caused by Metarhizium anisopliae}

The present invention relates to a composition and feed composition that can treat or prevent fungal diseases of white-spotted flower radish, preferably white-spotted radish green-gang disease, containing microorganisms of the genus Pediococcus.

White-spotted flower radish ( Protaetia brevitarsis ) is an insect belonging to the Coleoptera order and is widely distributed in temperate regions such as Korea, China, East Asia, and Europe. During the larval stage, it uses dead trees, compost, and humus rich in organic matter as a food source, and lives there until it becomes a pupa. In Korea, the larvae of the white-spotted daisy have long been called white grubs, and they are one of the most commonly used insects for food and medicinal purposes. Larvae are known to be effective in treating stomatitis, tetanus, and stroke, and various studies have proven their effects on infection, cirrhosis, fatigue recovery, and anticancer effects. The white-spotted radish larvae, along with rhinoceros beetle larvae, white river beetles, silkworms, grasshoppers, brown mealworm larvae, and double-star crickets, were registered as general food ingredients through certification by the Ministry of Food and Drug Safety in 2016. Interest in and demand for white-spotted lotus larvae is increasing. The number of white-spotted daisy farms in Korea is steadily increasing.

In most farms, larvae are raised in large quantities in small spaces to increase space efficiency when raising insects. This rearing method provides an environment vulnerable to the occurrence of insect pathogenic diseases, and mass mortality due to insect pathogenic diseases that occurs during larval rearing reduces larval productivity of rearing farms. There are about 700 species of entomopathogenic fungi that cause disease in insects, and Beauveria is a representative example. bassiana , Metarhizium anisopliae , Lecanicillium lecanii , etc. Among these, Metarhizium anisopliae ) is a fungus that causes rust disease and is widely distributed worldwide and lives on the surface of plant leaves and soil. Green liver disease is infected when larvae eat food or through contact with the epidermis, and is one of the most common diseases when rearing white-spotted radish larvae.

Accordingly, there is a need for the development of a new control composition that can control rust disease that occurs in the rearing of white-spotted radish larvae, while not affecting the growth of or enhancing the growth of white-spotted radish larvae.

The problem to be solved by the present invention is to provide a composition for controlling fungal disease of white-spotted radish, containing microorganisms of the genus Pediococcus.

Another problem to be solved by the present invention is Metarhizium using a composition for controlling fungal disease of white-spotted radishes containing microorganisms of the genus Pediococcus. anisopliae ) to provide a control method for rust-derived rust disease.

Another problem to be solved by the present invention is to provide a feed composition for promoting the growth of white-spotted flower radish containing microorganisms of the genus Pediococcus.

Another problem that the present invention aims to solve is to provide a method for controlling the population of white-spotted daisy using microorganisms of the genus Pediococcus.

In order to solve the above problems, the present invention provides a composition for controlling fungal disease of white-spotted radish, which includes Pediococcus spp. microorganisms or their culture medium.

White-spotted radish fungal disease is an insect disease caused by infection with mold (fungus). The white-spotted radish fungal disease of the present invention preferably refers to green-spotted radish fungal disease caused by infection with Metarhizium anisopliae . do. Green muscardine is a type of sclerosis in which insect larvae harden. It is transdermally infected by green muscardine bacteria. Large brown and black lesions are produced on the infected insects, which harden and harden after death. It is a disease that causes large numbers of bright green conidia to form on the body surface. Because the green conidia infect other larvae as a new source of infection, control of green liver disease is a very important task in insect farming.

In the present invention, the composition containing the Pediococcus genus microorganism or its culture medium is Pseudomonas ( Metarhizium anisopliae ) and exhibits antifungal activity.

In the present invention, the antifungal composition may have the same meaning as antibiotic, which is a general term for antimicrobial agents, and refers to a substance that can inhibit, inhibit, or kill the growth and life functions of Pseudomonas bacteria.

As used herein, “control” refers to the administration or application of a therapeutic agent to a subject or the performance of a procedure or manner in a subject for the purpose of obtaining a therapeutic benefit for a disease or related condition caused by a fungus. “Prophylaxis,” “preventing,” and the like refer to approaches for preventing, inhibiting, or reducing the likelihood of occurrence or recurrence of a disease or condition. It also refers to delaying the onset or recurrence of a disease or condition or delaying the development or recurrence of a disease or condition.

According to the present invention, the composition for controlling fungal disease in white-spotted radish, containing microorganisms of the Pediococcus genus or a culture medium thereof, can be used to prevent larvae from being infected with Metarhizium anisopliae as well as larvae of white-spotted radish, which can cause rust disease. It is a control composition applicable to all insect larvae. Examples of insect larvae that can cause rust disease include the larvae of insects belonging to the order Coleoptera , the larvae of insects belonging to the order Lepidoptera , the larvae of insects belonging to the order Hymenoptera , and the larvae of insects belonging to the order Orthoptera . It may be any one or more selected from the group consisting of larvae, and examples of insect larvae in which rust disease can occur are not limited to these.

According to the present invention, the microorganism of the Pediococcus genus may be a strain of Pediococcus pentosaceus, and preferably Pediococcus pentosaceus (accession number KACC 12311). It can be. According to the present invention, administration or treatment of the Pediococcus genus strain or its culture medium can effectively control mycosis without causing toxicity to white spotted radish larvae.

According to the present invention, the composition for controlling fungal disease in white spotted flower radish can suppress the development of rust disease in white spotted flower radish larvae and reduce the mortality rate caused by infection with rust bacteria. In one embodiment of the present invention, treatment with the composition for controlling fungal disease of white-spotted radish inhibited the reproduction of Pseudomonas in larvae infected with Pseudomonas, reduced hardening, and decreased the mortality rate of larvae.

According to the present invention, the composition may be fed to white spotted radish larvae or provided in a rearing environment. The feeding may be provided together with or separately from the larvae's food.

According to the present invention, the composition for controlling fungal disease in white-spotted radish can suppress the development of green-gold disease in white-spotted radish larvae while simultaneously increasing their body weight. In one embodiment of the present invention, treatment with the composition for controlling fungal disease of white-spotted radish radish reduced the occurrence of green-gold disease and increased the body weight of the larvae.

According to the present invention, the composition for controlling fungal disease in white-spotted flower radish can suppress the development of rust disease in white-spotted flower radish larvae, while at the same time shortening the larval period of the larvae and promoting growth and early pupation. In one embodiment of the present invention, treatment with the composition for controlling fungal disease of white-spotted radish radish reduced the occurrence of green-gold disease, while the larval period was shortened compared to the control group, and early pupation progressed.

According to the present invention, the growth enhancement may include shortening the larval period, increasing the weight of the larvae, rapid pupation and improvement of the pupation rate, rapid emergence and improvement of the emergence rate. The composition for controlling fungal disease of white-spotted flower radish according to the present invention prevents the occurrence of green-gold disease and induces rapid growth into adults.

Compositions of the present invention may be formulated. The dosage form is preferably any one selected from the group consisting of wettable powder, liquid, emulsion, coating agent, smoker, granule, powder, granule, capsule, fine powder, coating agent, smoke film, and solid agent, and more preferably liquid or emulsion. , it is any one selected from the group consisting of coating agents, smoking agents, granules and solid agents, and most preferably it is any one selected from the group consisting of smoking agents, liquids and coating agents. In order to manufacture such various preparations, solvents or additives commonly used in the preparation of the composition may be used, and the concentration of the composition is used in an amount to effectively achieve the purpose of action. At this time, the specific concentration is in the form of the preparation. and may vary depending on the method.

Dosage or dosage will vary depending on the severity of the pathology and the judgment of the operator. Dosage or dosage decisions based on these factors are within the level of one skilled in the art, and amounts generally range from 0.001 mg/kg/day to approximately 2000 mg/kg/day. It may be administered or treated once, or it may be administered or treated several times. The above dosage or treatment amount does not limit the scope of the present invention in any way.

In addition, the present invention is Metarhizium Anisopliae )-derived rust control method, which method is used in the Pediococcus genus ( Pediococcus) . spp . ) Spraying a composition for controlling fungal disease of white-spotted radish, containing microorganisms or their culture medium, into the breeding environment of insect larvae; Mix with food and feed to insect larvae; It provides a method for controlling rust disease derived from Metarhizium anisopliae , which includes providing insect larvae with one or more selected from the group consisting of direct spraying on insect larvae.

According to the present invention, Pediococcus genus spp . ) Microorganisms are the same as presented previously.

By spraying the composition for controlling fungal disease of white-spotted radish according to the present invention in advance into the breeding environment of insect larvae, before rearing, Metarhizium anisopliae ) can provide an environment in which they do not exist.

In addition, the present invention relates to the genus Pediococcus spp . ) Provides a feed composition for promoting the growth of white-spotted flower radish containing microorganisms or their culture medium.

According to the present invention, Pediococcus genus spp . ) Microorganisms are the same as presented previously.

Pediococcus genus according to the present invention spp . ) A composition containing microorganisms or their culture medium can increase the weight of white-spotted radish larvae and shorten the larval period, so it is effective as a feed composition for promoting the growth of white-spotted radish larvae.

In the present invention, the feed may be feed for insects, fish, birds or mammals, but is not limited thereto, and preferably has wild habits as defined in Article 2, Paragraph 1 of the Livestock Act and Article 2 of the Enforcement Rules of the same Act. It is purified and suitable for breeding and can be feed for livestock or insects that can contribute to increasing the income of farmers. The livestock may include cattle, horses, mules, donkeys, goats, goats, sheep, deer, pigs, rabbits, poultry, etc., and the poultry may include chickens, turkeys, ducks, ostriches, geese, quails, etc., preferably chickens. It is not limited to this as long as it is suitable for breeding and obtaining livestock products. The above “livestock products” are defined by Article 2, Paragraph 3 of the Livestock Act, as defined by the Ordinance of the Ministry of Agriculture, Food and Rural Affairs, as meat, milk, eggs, honey and their processed products, raw skins (including raw fur), raw wool, and other livestock products produced from livestock. means that Additionally, it may be a dog, cat, etc., including companion animals, but is not limited thereto.

The term "feed" of the invention means any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals. In one embodiment, the Pediococcus genus of the present invention ( Pediococcus spp . ) Feed compositions containing microorganisms or their culture fluid may include concentrated feed, roughage and/or special feed.

Concentrated feed includes seeds and fruits, including grains such as wheat, oats, and corn; bran, which includes rice bran, bran, and barley bran, which are by-products obtained from refining grains; soybeans; oil, sesame seeds, linseed, and coco oil; Fish soluble, meat powder, which is a concentrated product of fresh liquid obtained from fish meal, fish waste, fish meal, fish waste, etc., such as residual starch, which is the main component of the starch residue that is left after removing the starch from the by-products such as seed jelly, sweet potatoes, potatoes, etc. , animal feeds such as blood meal, feather meal, skim milk powder, dried whey, which is the residue from milk to cheese, and casein from skim milk, yeast, chlorella, seaweed, etc.

Forage includes raw grass feed such as wild grasses, grasses, and green cuttings, turnips for feed, beets for feed, root vegetables such as rutterbearger, a type of turnip, raw grass, green cuttings, and grains, etc., in silos. There are silage, a stored feed that is stuffed and fermented with lactic acid, field grass, hay made by cutting and drying grass, straw from crops for breeding livestock, and leaves from legumes. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea and its derivative diureide isobutane, and mixed feeds to supplement ingredients that are likely to be lacking when mixing only natural feed raw materials, or to increase the storability of the feed. There are feed additives, which are substances added in trace amounts.

In addition, the present invention is a method of controlling the population of white-spotted radish,

When the demand for white spotted flower radish increases than the supply, the larvae of the white spotted flower radish are fed to the genus Pediococcus . spp . ) Shorten the larval period of white-spotted radish by feeding microorganisms or their culture medium,

When the demand for white spotted flower radish decreases than the supply, white spotted flower radish larvae are fed to the genus Pediococcus . spp . ) Provides a method for controlling the population of white-spotted radish, including excluding feeding of microorganisms or their culture medium.

According to the present invention, the feeding may be provided together with the feed of white-spotted radishes or provided in a breeding environment for white-spotted radishes.

The population control of white-spotted daisy according to the present invention can be easily applied to overcome the problem of oversupply or shortage of white-spotted radish that may occur in insect breeding farms and to control shipment volume.

Pediococcus genus according to the present invention spp . ) A composition containing microorganisms or their culture medium suppresses the development of rust disease caused by Metarhizium anisopliae infection from white-spotted radish larvae, not only lowers the mortality rate of larvae, but also increases the weight of larvae and prolongs the larval period. It can improve growth by shortening the length. Therefore, the genus Pediococcus spp . ) Compositions containing microorganisms or their cultures are useful as compositions for controlling rust diseases, such as white-spotted radish mycosis, and are also useful as feed compositions for promoting the growth of white-spotted radishes. In addition, Pediococcus genus according to the present invention spp . ) The use of a composition containing microorganisms or their culture medium can control the larval period in the breeding of white-spotted daisy, so that the population of white-spotted daisy can be controlled without the problem of toxic ingredients or stress, and can be used in industrial sites. It can be usefully applied.

Figure 1 shows the results of confirming the occurrence of rust disease caused by treatment with Pediococcus spp . microorganisms in white-spotted flower radish larvae treated with rust bacteria through the larval mortality rate.
Figure 2 shows the appearance of white spotted radish larvae according to the presence or absence of rust disease. Figure 2a is an image of a normal uninfected larva, and Figure 2b is an image of a larva with green steel disease.

Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are merely illustrative of the present invention, and it is clear to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the attached patent claims.

Experimental strain

Metarhizium, the causative agent of rust disease, was used. anisopliae ) was distributed and used from the Korean Agricultural Culture Collection (KACC) of the National Institute of Agricultural Sciences (KACC 40969). The cultured Pseudomonas bacterium was cultured on SDAY (Sabouraud Dextrose Agar Yeast) medium and incubated at 25 ± 2°C for more than 3 weeks. After culturing, the formation of spores was checked and then used in the experiment. When mold spores were formed, they were diluted in 0.02% tween 20 and used as a spore suspension.

Pediococcus genus strain was Pediococcus pentosaceus (accession number KACC 12311) distributed from the Microbial Bank of the National Institute of Agricultural Sciences.

experimental insects

The larvae of the white-spotted marigold were purchased from the Insect Industry Department of the National Institute of Agricultural Sciences and used. To test the effect of Pediococcus strains on rust disease that occurs when raising white-spotted daisy, early 3rd instar larvae were used, and larvae with uniform weight (1.1-1.2 g) were selected. The larvae were reared under conditions of 25°C temperature, 60% relative humidity, and 60% moisture content of sawdust. Fermented sawdust (Gum Life) and 10 white-spotted radish larvae were reared in a rectangular rearing container (210*120*130 mm).

data analysis

To test the effect of strains of the genus Pediococcus on inhibiting the development of rust disease and increasing body weight gain against rust disease that occurs when raising white-spotted daisy, the significance of each mean was compared through one-way ANOVA analysis and t-test (PAST) , version 2.17c (Hammer et al., 2001). Significance differences between groups were verified at the p<0.05 level using Tukey's HSD test. In the results, the mortality rate of white-spotted larvae was expressed as mean ± standard error (SE), and the body weight was expressed as mean ± standard deviation (SD).

Example 1

The third instar larvae of White-spotted Japanese Radish were treated with KACC, a strain of the genus Pediococcus. The treatment method was to mix the strain of the genus Pediococcus with tertiary distilled water and mix it with dried fermented oak sawdust to adjust the moisture content of the sawdust to 60% before feeding. . Pseudomonas spore suspension was treated at a concentration of 1 × 10 ⁷ conidia/ml per 100 g of oak fermented sawdust, and the control group was treated with 0.02% tween 20. The mortality rate of larvae due to rust disease was confirmed for 10 weeks after mold treatment. Infection with Pseudomonas disease was determined if Pseudomonas spores were formed on the epidermis of dead larvae, indicating death by Pseudomonas bacteria. The experiment was conducted at one-week intervals, and additional sawdust for each treatment group was added at two-week intervals after the start of the experiment, after removing feces from the existing sawdust. The experiment was repeated five times for the control group and the experimental group, respectively.

Test example One. pediococcus by genus strain treatment rust disease Outbreak containment assessment

Suppression of the development of rust disease in white-spotted radish larvae was confirmed by evaluating the change in mortality rate of larvae and the start date of mortality according to the presence or absence of treatment with strains of the Pediococcus genus, strain species, and treatment period.

Figure 1 shows the results of confirming the inhibition of rust disease development by treatment with strains of the Pediococcus genus in terms of larval mortality. The gray line is the control, untreated group (0%), and the blue line is the mortality rate (%) of the group treated with KACC 12311. )am. The untreated group showed a high mortality rate of about 70%, but treatment with Pediococcus strains appeared to lower the mortality rate caused by Pseudomonas bacteria. In particular, the mortality rate of the group treated with KACC 12311 was 16.67%.

Test Example 2. Appearance evaluation of green steel disease larvae

Changes in the appearance of the third instar larvae of white-spotted radish performed in Example 1 were evaluated with the naked eye according to the presence or absence of rust infection, and this is shown in Figure 2. Figure 2a is an image of a normal uninfected larva (control), and Figure 2b is an image of a larva infected with rust disease. It was observed that the white-spotted radish larvae infected with rust disease had a stiff appearance and their epidermis was covered with dark green spores.

Claims (12)

A composition for controlling rust disease caused by Metarhizium anisopliae , comprising a Pediococcus spp. microorganism or a culture medium thereof. According to paragraph 1,
A composition for controlling rust disease on white-spotted flower stalks caused by Metarhizium anisopliae , where the microorganism of the Pediococcus genus is Pediococcus pentosaceus strain (accession number KACC 12311). delete According to paragraph 1,
A composition for controlling rust disease in white spotted flower radish caused by Metarhizium anisopliae , which suppresses the development of rust disease in white spotted flower radish larvae and lowers the mortality rate due to infection by rust bacteria. According to paragraph 1,
A composition for controlling rust disease in white spotted flower radish caused by Metarhizium anisopliae , which suppresses the development of rust disease in white spotted flower radish larvae and increases body weight. According to paragraph 1,
The composition is a composition for controlling white-spotted cauliflower larvae or provided in a rearing environment, which is caused by Metarhizium anisopliae . Metarhizium anisopliae )-derived rust disease control method, the method is
Spraying a composition for controlling fungal disease of white-spotted radish, containing Pediococcus spp. microorganisms or their culture medium, into the breeding environment of insect larvae; Feeding insect larvae; And direct spraying on insect larvae; comprising providing white-spotted radish larvae with one or more methods selected from the group consisting of: Metarhizium Anisopliae )-derived rust control method. In clause 7,
The microorganism of the Pediococcus genus is Pediococcus pentosaceus strain (accession number KACC 12311) 9, method for controlling rust disease. In clause 7,
The insect is one or more selected from the group consisting of Coleoptera insects, Lepidoptera insects, Hymenoptera insects, and Orthoptera insects. Pediococcus pentosaceus pentosaceus ) strain (accession number KACC 12311) or a culture medium thereof. A feed composition for promoting the growth of white-spotted radish. According to clause 10,
A feed composition for promoting the growth of white-spotted radish larvae, which increases the weight of larvae and shortens the larval period. As a method of controlling the population of white-spotted daisy,
When the demand for white-spotted daisy increases than the supply, the larvae of the white-spotted daisy are infected with Pediococcus pentosaceus. pentosaceus ) strain (accession number KACC 12311) or a composition containing its culture medium to shorten the larval period of white-spotted radish,
When the demand for white-spotted daisy decreases than the supply, Pediococcus pentosaceus Pentosaceus ) strain (accession number KACC 12311) or a culture medium thereof.