KR20150101702A - Composition for having effect on suppression of bacterial soft rot disease and control method for the same - Google Patents

Abstract

The present invention relates to an agent for controlling bacterial soft rot and a control method thereof and, more specifically, to a culture composition for culturing a Paenibacillus genus ASU-64 strain which is a novel separation strain having antimicrobial activity with respect to bacterial soft rot Erwinia which is a bacterial pathogen plant, and to a control method of bacterial soft rot Erwinia using the same.

Description

Technical Field [0001] The present invention relates to a Chinese cabbage herbal roots disinfectant and a method of controlling the same. BACKGROUND ART [0002]

More particularly, the present invention relates to a culture composition obtained by cultivating a strain of the genus Penny Bacillus ASU-64, which is a newly isolated strain having antimicrobial activity against Erwinia , a plant bacterial pathogen, Used cabbage rolls Erwinia And a method of controlling the same.

Cabbage is a low temperature vegetable that likes a cool climate. When it reaches high temperature in the growing season, it causes damage to various diseases and pests. Among them, the Chinese cabbage blossom that occurs during the cabbage growing period penetrates directly through the surface of the plant from the surface of the plant near the root and spreads to the whole under the cabbage or the whole cabbage, causing the cabbage to rot and causing serious damage to the entire cabbage field.

 The cabbage blight is a major limitation in the production of summer cabbage, and it causes great damage during transportation, sale and storage, and not only cabbage but also cabbage, celery, radish, tobacco, onion, tomato, carrot, potato, lettuce or wasabi It is also a serious disease that causes damage to various major vegetables.

In addition, the Chinese cabbage blight disease is a bacterial and soil-borne plant disease, and the bacterial disease has a very low control effect of the medicine, so that it is difficult to prove effective if a large amount of medicines are not sprayed prophylactically. However, antibiotics such as streptomycin, which have been used for the control of bacterial diseases such as Chinese cabbage blight, are expensive and can cause resistant bacteria. In case of using a copper disinfectant, metal contamination may occur in the soil and the target plants There was a problem.

On the other hand, the control method using bio-pesticides has little toxicity to safety of environment and to produce, and it can minimize the problem of ecosystem disturbance because microorganisms commonly used in nature are used.

As a method for solving the above-mentioned problems, a prior art related to a microorganism for control of Chinese cabbage rot disease which is conventionally present will be briefly described.

First, Korean Patent No. 0580336 (2006.05.09) relates to a new Bacillus sp. Microorganism having the effect of controlling the Chinese cabbage blight. According to this, Bacillus sp. sp . ) 53 strain is effective as a pesticide for preventing bacterial blight of cabbage blight due to its excellent controlling effect against bacterial blight of cabbage blight.

Korean Patent No. 1206883 (November 26, 2012) relates to a novel strain of Streptomyces sp. Having an appropriate density recognition inhibitory activity. Specifically, Streptomyces AN090072 and Streptomyces strain AN090571, which have a bacterial proper density recognition-inhibitory activity, are Escherichia Coli), Bacillus subtilis (Bacillus The evaluator has confirmed that it has an subtilis) and Salmonella typhimurium grains Titanium (effect of inhibiting the rot caused by the air Winiah Caro sat see (Erwinia carotovora) even in cabbage do not show antibacterial activity against pathogenic bacteria of the Salmonella typhimurium), a variety of bacterial organisms Lt; RTI ID = 0.0 > infectious < / RTI >

Korean Patent Registration No. 0743672 (Jul. 23, 2007) relates to the pyrrolidine A1 having competitive inhibitory activity of acyl homoserine lactone. More particularly, the present invention relates to a kitasatospora genus AR030054 ( Kitasatospora ) having competitive inhibitory activity of acyl homoserine lactone, a quorum sensing signaling substance, sp . AR030054) [KCTC10904BP], which can be used for the treatment or prevention of infectious botanical diseases caused by various bacterial organisms.

Korean Patent No. 0751448 (Aug. 16, 2007) relates to bacterial strains, genes and enzymes that regulate bacterial diseases by eliminating the quorum sensing signal, and more particularly, to a method for coding bacterial metabolism regulators Lt; RTI ID = 0.0 > bacterial < / RTI > diseases.

Korean Patent No. 1010537 (Jan. 18, 2011) relates to a novel Lactobacillus KLF01 and Lactococcus LKCO2 species and a composition for preventing plant disease using the same, and relates to a novel lactose bacterium having excellent control effect against bacterial blight, Bacillus KLF01 ( Lactobacillus KLF01) and Lactococcus KLC02 ( Lactococcus KLC02).

Studies on the development of Chinese cabbage herbicide control agents using microorganisms such as Bacillus, Lactobacillus, Lactococcus and Streptomyces spp. And control of Chinese cabbage rot by controlling the quorum and transgenic transformation of Chinese cabbage as described above Is now underway.

According to the present invention, a novel strain controlling the Erwinia mildew of the Chinese cabbage strain was isolated and identified as Pennybacillus strain, and the strain Pennibacillus genus ASU-64 ( Penibacillus sp. ASU-64). Thus the invention cabbage soft rot of Chinese cabbage using penny Bacillus having antimicrobial activity of the Erwinia ASU-64 strain cultured composition and this soft rot Erwinia I would like to suggest a control method.

DISCLOSURE Technical Problem The present invention has been conceived to solve the above-mentioned problems, and an object of the present invention is to provide a culture composition in which a strain of Penny Bacillus genus ASU-64, which is a novel strain having antibacterial activity, is cultivated in Chinese cabbage Erwinia .

It is another object of the present invention to provide a method for controlling a Chinese cabbage herb controlling agent to soil or a plant through various formulation methods using the strain of the genus Penny Bacillus ASU-64 and its culture composition.

The Chinese cabbage herb remedy according to an embodiment of the present invention is characterized by comprising a strain of Penibacillus sp. ASU-64 (accession number: KCTC 12546BP) of the genus Penny Bacillus.

In addition, the Chinese cabbage herb remedy according to an embodiment of the present invention is characterized in that the culture supernatant obtained by culturing the strain of the genus Penny Bacillus ASU-64 is diluted 1 to 1000 times.

In addition, in the Chinese cabbage herbicide controlling agent according to an embodiment of the present invention, the strain of the genus Penny Bacillus ASU-64 can be cultured in the presence of β-Cyclodextrin, Dextrin, Glycogen, Arbutin, D-Cellobiose, D- D-Glucoside, D-Raffinose, Salicin, Stachyose, or D-Trehalose as a carbon source.

In addition, in the Chinese cabbage herb composition according to the embodiment of the present invention, the composition for controlling the Chinese cabbage herb disease may be selected from the group consisting of cabbage, celery, radish, tobacco, onion, tomato, carrot, potato, lettuce, And has a controlling effect on blistering which occurs in the hair.

According to another embodiment of the present invention, there is also provided a method for controlling the Chinese cabbage rot disease, comprising the steps of : isolating Penicillus sp. ASU-64 ( Penibacillus sp. sp. ASU-64) (Accession No .: KCTC 12546BP), which is then used as an eco-friendly plant preparation.

According to another embodiment of the present invention, there is provided a method of controlling Chinese cabbage blight, wherein the culture supernatant obtained by culturing the strain of the genus Penny Bacillus ASU-64 is diluted 1 to 1000 times and sprayed on a crop or soil.

The present invention also provides a method of controlling Chinese cabbage blossom according to another embodiment of the present invention. The method of the present invention is a method for controlling Chinese cabbage rot disease, comprising the steps of: Characterized in that said strain of the genus Penny Bacillus ASU-64 is cultivated using as a carbon source D-mannitol, D-mannose, D-Melibiose,? -Methyl D-Glucoside, D-Raffinose, Salicin, Stachyose or D-Trehalose.

According to another embodiment of the present invention, the culture supernatant obtained by culturing the strain of the genus Penny Bacillus ASU-64 is not only the cabbage but also cabbage, celery, radish, tobacco, onion, tomato, carrot , Potatoes, lettuce or horseradish.

The present invention relates to a Chinese cabbage herbicide controlling agent and a method of controlling the same, and as a result of an experiment of cultivating a culture of a new strain of the genus Pennibacillus ASU-64 as a Chinese cabbage herbicide, And it was proved to be a species capable of effectively controlling cabbage rot disease.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing an identification result of a strain of the genus Penny Bacillus ASU-64 according to an embodiment of the present invention. FIG.
FIG. 2 is an exemplary diagram showing the results of culture of a Penny Bacillus sp. ASU-64 strain according to a carbon source according to an embodiment of the present invention. FIG.
FIG. 3 is an exemplary view showing a carbon source of a strain of the genus Penny Bacillus ASU-64 according to an embodiment of the present invention.
FIG. 4 is an exemplary view showing the antimicrobial activity against the Erwinia cabbage strain of Penny Bacillus sp. Strain ASU-64 according to an embodiment of the present invention.
FIG. 5 is a graph showing an inhibitory effect on the Erwinia herbaceous herb according to the dilution ratio of the culture supernatant of the strain Penny Bacillus sp. ASU-64 according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited to these Examples.

- Example 1-

Detailed description will be given on the isolation and selection of bacteria showing antimicrobial activity against Erwinia .

Generally Recognized As Safe (GRAS) samples were suspended and diluted in sterilized physiological saline to inoculate the microorganisms showing antimicrobial activity against Erwinia , a Chinese cabbage strain, and then inoculated into LB agar medium and incubated at 30 ° C for 2 days To isolate microorganisms.

In order to isolate and isolate strains which have antimicrobial activity in Erwinia , a strain of pure isolates, the strains exhibiting antibacterial activity were selected using the culture supernatant of each strain.

As a result of identification, 16S rRNA sequencing was performed to identify bacteria having antibacterial activity. As shown in Fig. 1, the strain was identified as Pennybacillus sp. And the strain Penny Bacillus sp. ASU-64 ( Penibacillus sp. sp. ASU-64). In addition, the above strain of Penny Bacillus genus ASU-64 was deposited with the Korea Research Institute of Bioscience and received the deposit number KCTC 12546BP.

- Example 2-

The types of carbon sources necessary for culturing the above-mentioned strain of the genus Pennibacillus ASU-64 will be described.

As shown in FIG. 2, in the present invention, Biolog Plate GN2 was used to investigate the type of carbon source of the strain of the genus Penny Bacillus ASU-64. The strain of Penny Bacillus genus ASU-64 was adjusted to OD (Optical Density) of 0.3 in a salin buffer using the above Biolog plate GN2, and was inoculated into 96 wells at a rate of 150 ul (microliter). Positive.

FIG. 3 is a graph showing the carbon source of the Penny Bacillus sp. Strain ASU-64 which was confirmed to be purplish in the experiment shown in FIG. 2 as a positive result (positive: gray).

As shown in FIG. 3, the carbon source of the Penny Bacillus sp. ASU-64 strain, which turned purple as a result of the culture, was found to be β-Cyclodextrin, Dextrin, Glycogen, Arbutin, D-Cellobiose, D- maltotriose, D-mannitol, D-mannose, D-Melibiose,? -Methyl D-Glucoside, D-Raffinose, Salicin, Stachyose or D-Trehalose.

- Example 3-

About Erwinia The culture conditions and antimicrobial activity of the strain Penny Bacillus sp. ASU-64 having controllability are described.

Tryptone 1%, Glucose 2%, Yeast extract 0.5% , and NaCl in the penny Bacillus 1% in the added medium ASU-64 strain in 30 ℃ 3 days after the culture the culture supernatant the commonly known disk for cabbage soft rot Erwinia The antimicrobial activity was confirmed by the diffusion method. That is, 100 ul of the culture supernatant of the strain of the genus Penny Bacillus ASU-64 was loaded on each test disc 20, and after drying, the antibacterial activity was examined by placing on the medium 10 on which the Erwinia vinegar was grafted.

As a result, as shown in FIG. 4, it was confirmed that a growth inhibition ring 30 was formed around the test disc 20 loaded with the culture supernatant of the strain of the genus Penny Bacillus ASU-64. As a result, the strain of the genus Penny Bacillus ASU-64 was found to exhibit an antibacterial activity against Erwinia rotiflora .

- Example 4-

Pine berry seedlings using culture supernatant of ASU-64 strain of Bacillus genus Erwinia The control method will be described.

FIG. 5 is a graph showing the results of experiments showing the inhibitory effect on the Erwinia herbaceousum according to the dilution ratio of the culture supernatant of the strain Penny Bacillus sp. ASU-64 according to an embodiment of the present invention.

As shown in FIG. 5, experiments were conducted to confirm the inhibitory effect of the strain of the genus Penny Bacillus ASU-64 on Chinese cabbage blight. For reference, Erwinia was used to induce vegetative bacterial pathogen causing Chinese cabbage blight and Chinese cabbage was used for the crop.

Each of 5 leaves of cabbage was wounded, and one of them was inoculated by adjusting the Erwinia culture medium to OD 0.1 as a control. In addition, the remaining 4 specimens were inoculated with 100ul each of the culture supernatant of the genus Penny Bacillus, the 10-fold diluted solution, the 100-fold diluted solution, the 1000-fold diluted solution and the Erwinia culture solution at 30 ° C for 2 days.

The occurrence of Chinese cabbage blight was visually observed, and it was judged that Chinese cabbage blight occurred when the test site of cabbage leaves became cloudy after one day of experiment and changed into dark brown.

As a result of comparing the control group and the experimental group after the above experiment, it was found that in all the four experimental groups inoculated with Erwinia culture supernatant of the culture supernatant of the strain of Penny Bacillus sp. ASU-64, The highest degree of milky lotion inhibition was observed in the culture supernatant diluted with 1000 times diluted stock solution of Bacillus sp. ASU-64 strain supernatant.

This is in agreement with the fact that the greenhouse is generally diluted 1,000 times with the environment to treat the crops. Therefore, the efficacy of the above culture supernatant of the genus Penny Bacillus ASU-64 is expected to be excellent.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation. I will understand that. Accordingly, the technical scope of the present invention should be defined by the following claims.

10: Badge smear of cabbage milkshake ERWINIA
20: Test disc 30: Growth inhibition

Korea Biotechnology Research Institute KCTC12546BP 20140205

Claims (8)

(Penicillium sp. ASU-64) strain (Accession No .: KCTC 12546BP). The method according to claim 1,
Wherein the culture supernatant obtained by culturing the strain of the genus Penny Bacillus ASU-64 is diluted 1 to 1000 times. The method according to claim 1,
The strain of the genus Pennibacillus, ASU-64,
D-mannose, D-Melibiose, β-Methyl, D-galactose, α-D-glucose, maltose, maltotriose, D-mannitol, D-Glucoside, D-Raffinose, Salicin, Stachyose or D-Trehalose as a carbon source. The method according to claim 1,
The composition for controlling the Chinese cabbage blight disease,
The composition for controlling Chinese cabbage herbaceous diseases as set forth in claim 1, wherein the composition has an anti-acne effect on not only cabbage, but also cabbage, celery, radish, tobacco, onion, tomato, carrot, potatoes, lettuce or vegetables including vegetables. (Penicillus sp. ASU-64) strain (Accession No .: KCTC 12546BP) having antimicrobial activity against Chinese cabbage blight and cultivating the strain as an eco-friendly plant preparation. The method of claim 5,
Wherein the culture supernatant obtained by culturing the strain of the genus Penny Bacillus ASU-64 is diluted 1 to 1000 times and sprayed on the crop or soil. The method of claim 5,
β-Cyclodextrin, Dextrin, Glycogen, Arbutin, D-Cellobiose, D-Fructose, D-Galactose, α-D-Glucose, Maltose, Maltotriose, D-Mannitol, D-Mannose, , D-Raffinose, Salicin, Stachyose or D-Trehalose as a carbon source, and culturing the strain of the genus Penny Bacillus ASU-64. The method of claim 6,
The culture supernatant obtained by cultivating the strain of the genus Penny Bacillus ASU-64 is characterized by controlling not only the above-mentioned Chinese cabbage but also blight of vegetables including cabbage, celery, radish, tobacco, onion, tomato, carrot, potato, lettuce or horseradish As a control method for cabbage rot disease.

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