KR102252856B1 - Natural Plant Protection Agents Comprising Bacillus velezensis KCCM12751P Having Antifungal Activity Against Pathogens And Plant Growth Promoting Effect, And Uses Thereof - Google Patents

Abstract

The present invention relates to a Bacillus Velezensis HY3479 (Accession number: KCCM12751P) strain or a method for culturing the strain. By culturing in a vegetable medium (rice water medium containing crab shell powder) developed by itself by using waste resources from agriculture, forestry, agriculture and fisheries, the strain can be usefully used for controlling various plant pathogens and accelerating the growth of crops.

Description

Natural Plant Protection Agents Comprising Bacillus velezensis KCCM12751P Having Antifungal Activity Against Pathogens And Plant Growth Promoting Effect, And Uses Thereof}

The present invention relates to a novel Bacillus velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain or a microbial preparation containing the same, which has an effect of promoting plant growth and controlling plant diseases, and more specifically, to a vegetable medium (RCM; Rice washing water contained Crab Media; barley using a composition in which Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) was cultured in rice washing water contained Crab Media; Bacillus belegensis, a new microorganism that promotes plant growth of oats, peppers, and cabbages, and controls plant diseases against capsicum scleroderma, capsicum anthracnose, apple tree blight, cucumber seedling disease, ginseng root rot, strawberry gray fungus, and grass sheath blight. It relates to a HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain or a microbial preparation containing the same as an active ingredient.

Due to the recent expansion of the eco-friendly agricultural area, public interest in eco-friendly agriculture and eco-friendly agricultural products is increasing. In particular, the consumption of eco-friendly agricultural products is increasing rapidly due to the changing public perception of health and well-being. Therefore, from 2019, the government has strictly applied the Pesticide List System (PLS) for agricultural products to reduce the use of pesticides and promote safe agricultural products to the public. For the above reasons, there is a trend to limit the use of pesticides around the world due to problems (soil and river pollution, pesticide residues in agricultural products, etc.) that have been used in the past. Therefore, an alternative alternative to these pesticides is eco-friendly biocontrol using microorganisms, natural enemies or plant extracts. In particular, biocontrol using microorganisms is a microbial pesticide that controls pathogens or pests using antibiotics or toxins produced by microorganisms, and a microbial fertilizer that protects plants and indirectly promotes the growth of plants by using the antagonistic action of the microorganism itself. Etc. Therefore, most recent studies on plant disease control effects are the development of eco-friendly microbial preparations using microorganisms having antagonistic effects against plant pathogens.

Currently, farmers appear to be uncomfortable and uncertain about existing commercially available microbial agents. As a result of a 2010 survey of 111 farm households by the Korea Rural Economic Research Institute, 1) the price is too high (35.6%); 2) uncertainty about quality and efficacy (29.7%); 3) Lack of information on use and management (19.2%); 4) It was concluded that it was difficult to use and manage, and that it was difficult to solve (14.6%) when a problem occurred.

The inventors of the present invention were conducting research on various physiological activities of antagonistic microorganisms and antagonistic microorganisms against pathogens that cause disease in plants, while the novel microorganism Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) is a variety of plant pathogens. It was confirmed that it exhibits a plant growth promoting effect as well as a control effect on. Thus, Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) was identified as an excellent natural plant protector having various physiological activities, completing the present invention.

[Patent Document 001] Republic of Korea Patent No. 2148396, Characteristics of Bacillus Bellesensis CE 100 and the Effect of the Culture Filtrate on the Control of Citrus Black Spot Disease [Patent Literature 002] Korean Patent No. 2015051, Microbial preparation for controlling plant diseases including Bacillus bellegensis G341 strain and Ricinibacillus spericus TC1 strain, and plant disease control method using the same [Patent Document 003] Republic of Korea Patent No. 1163986, Bacillus belegensys G341 strain and plant disease control method using the same [Patent Document 004] Korean Patent No. 1199931, A method for promoting nitrogen mineralization of organic fertilizers and controlling plant diseases using Bacillus Bellesensis KRICT934 strain [Patent Document 005] Republic of Korea Patent No. 2124060, Novel Bacillus Bellesensis MWS28 and a microbial control agent containing the same and a method for producing the same [Patent Document 006] Republic of Korea Patent No. 1089149, Bacillus belegensys S3-5 strain and plant disease control method using the same [Patent Document 007] Republic of Korea Patent No. 1845708, a novel microorganism Bacillus belegensys YP2 or a microorganism preparation containing the same [Patent Document 008] Korean Patent No.0781300, a microbial preparation for controlling strawberry wilt disease using a novel Bacillus microbial strain, and a method for producing the same [Patent Document 009] Republic of Korea Patent No. 2078252, Bacillus Bellesensis ARRI17 having antagonism against ginseng root rot and its use [Patent Document 010] Republic of Korea Patent No. 0976760, Bacillus bellegensis NSB-1 strain, anthrax control composition containing the same, and anthrax control method

[Non-Patent Document 001] Devi et al., Depiction of secondary metabolites and antifungal activity of Bacillus velezensis DTU001, Synthetic and Systems Biotechnology, 4, 2019, 142-149 [Non-Patent Document 002] Lee Sang-yeop et al., Biological Control of Mustard Powdery Mildew of Bacillus velezensis YP2, Journal of Pesticide Science, 20(4), 2016, 369-374 [Non-Patent Literature 003] Lee Sang-yeop et al., Biological Control of Bacillus velezensis CC112 Strain Against Ginseng Jaloc Disease, Journal of the Korean Mycology Society, 44(3), 2016, 176-183

The present inventors studied the cost, efficacy, and convenience of use among the above disadvantages. As a result, a novel microorganism Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number) having a control effect against various plant diseases or a plant growth promoting effect. : KCCM12751P) It is to provide convenience for price and use through the method of mass cultivation of the above strain by applying the strain identification and appropriate technology.

It is an object of the present invention to provide a microbial preparation containing the entrusted microorganism or the same having a control effect against various plant diseases. More specifically, pepper sclerotic disease (Sclerotinia sclerotiorum ), capsicum anthrax ( Coletotrichum acutatum ), apple tree blight (Phytophthora cactorum), cucumber ultimum (Phythium ultimum), tomato gray mold ( Botrytis cinerea ), red pepper sollock disease (Rhizoctonia) , And a new microorganism Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) or a microbial preparation containing it as an active ingredient, which has a control effect against plant pathogens selected from the group consisting of ginseng root rot ( Cylindrocarpon destructans). It is in providing.

The present invention provides a new strain Bacillus velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) for controlling plant diseases.

The present invention includes a novel microorganism Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain and a culture thereof.

In the configuration of the present invention, the Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) contains at least one or more of the bacterial cells, the culture medium, the culture filtrate as an active ingredient, and exhibits a plant disease control effect.

In the composition of the present invention, as a fraction obtained from the culture solution or culture filtrate of the Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain, at least one of an ethyl acetate layer, a butanol layer, and an aqueous layer is used as an active ingredient. Including plant growth promoting effect.

In the composition of the present invention, the plant disease, pepper sclerotic disease ( Sclerotinia sclerotiorum ), pepper anthrax ( Coletotrichum acutatum ), apple tree blight (Pytophthora cactorum ), cucumber ultimum (Phythium ultimum ), tomato gray mold ( Botrytis cinerea ), pepper It is characterized in that it is derived from one or more pathogens selected from the group consisting of Jaloc's disease ( Rhizoctonia solani ), and ginseng root rot ( Cylindrocarpon destructans).

In the composition of the control method of the present invention, at least one of seed coating, seed soaking, soil improvement, foliar spraying, and drench spraying is used using a natural plant protection agent.

In the composition of the control method of the present invention, the natural plant protection agent includes at least one or more of bacterial cells, culture medium, and culture filtrate of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain as an active ingredient. use.

In the composition of the control method of the present invention, it is characterized in that the natural plant protection agent is treated at a concentration of ^{10 5} to 10 ^{9 cfu/mL.}

In the composition of the control method of the present invention, it is treated on a plant selected from the group consisting of peppers, beans, barley, oats, cabbages, strawberries, cucumbers, ginseng and chives.

In the composition of the control method of the present invention, pepper sclerotic disease (Sclerotinia sclerotiorum ), pepper anthrax ( Coletotrichum acutatum ), apple tree blight (Pytophthora cactorum ), cucumber ultimum (Phythium ultimum ), tomato gray mold ( Botrytis cinerea ), red pepper cinerea ( Rhizoctonia solani ), and ginseng root rot ( Cylindrocarpon destructans ) characterized by exhibiting a control effect on at least one plant disease selected from the group consisting of.

The present invention provides a new soil-derived Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain.

The strain has an organic substance degrading enzyme (starch, protein, fibrin degrading enzyme) activity and excellent antifungal activity against various plant pathogens.

The microbial preparation of the present invention can be mass cultured in a non-sterile environment without separate culture.

The mass cultivation of the novel Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain of the present invention uses a self-developed vegetable medium (RCM; Rice water contained Crap Media), so it is inexpensive to farmers by reducing production cost. Since it can be supplied at a cost, there is an effect of reducing the cost of purchasing pesticides.

1 is a result of confirming the colony on the plate of a novel Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain of the present invention.
2 is a 16s-rRNA nucleotide sequence of a novel Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain.
3 is a result of measuring the growth of a new Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain in a vegetable medium (RCM, Rice water contained Crap Media).
FIG. 4 is a result of confirming the activity of an organic substance degrading enzyme of a novel Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain cultured in a vegetable medium (RCM).
5 is a result of confirming the antifungal activity against various plant diseases of a novel Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain cultured in a vegetable medium.
6A and 6B are results of confirming the antifungal activity against plant diseases in the organic solvent fraction of the novel Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain.
7 is a result of confirming the effect of promoting oat sprout growth and inhibiting fungi in the oat sprout of a novel Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain cultured in a vegetable medium.
8 is a result of a field pepper demonstration test.

Hereinafter, the present invention will be described in more detail through the following examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

At this time, unless there are other definitions in the technical terms and scientific terms used, they have the meanings commonly understood by those of ordinary skill in the technical field to which the present invention belongs, and the gist of the present invention in the following description and accompanying drawings Descriptions of known functions and configurations that may be unnecessarily obscure will be omitted.

(Example 1) New Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain isolation

The Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain used in this study was isolated by collecting soil from an eco-friendly agricultural complex of Pohang Nodaji Village Co., Ltd. The strain was streaked on tryptone soybean agar (TSA) to obtain pure colonies. Thereafter, the obtained culture was cultured in Tryptone soy broth (TSB) at 40° C. for 1 day at 200 rpm using a shaking incubator. The resulting bacterial culture was ^{adjusted to 10 8} CFUs/mL (CFU, colony forming units) and treated with 20% (v/v) glycerol for further experiments and stored frozen at -80°C (FIG. 1).

(Example 2) Identification of a new Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain

In order to identify the novel microbial strain isolated in the present invention, the base sequence was determined by amplifying 16s-rRNA. In order to amplify the nucleotide sequence, the analysis was conducted by requesting Microgen co., Ltd., Korea, and the results showed similarity to the Bacillus velezensis spp. Accordingly, the microbial strain isolated and identified according to the present invention was named Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) and was deposited as an international patent strain with the Korean Seed Association, and was given the accession number KCCM12751P ( Fig. 2).

(Example 3) Development of low-cost vegetable medium

Rice water contained Crap containing 0 to 2% of crab skin powder produced by Pohang Nodaji Village Co., Ltd. to mass-produce Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain at low cost. Powder Media (hereinafter referred to as RCM) or rice water medium (hereinafter referred to as RM) or a medium containing 2% rice powder (2% Rice powder Media, hereinafter referred to as RM_002) was developed and used for mass propagation of the strain. In more detail, Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) is a low-cost vegetable medium used for mass propagation culture using RM or a rice-water medium (RCM) containing 1% crab skin powder. Was used. Therefore, as a result of analyzing the 9 nutrients for the rice water medium (RM) and the medium containing 2% rice flour (RM_002), the protein was standardized in the vegetable medium of the Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain. In addition, as a result of analyzing the ingredients of the crab shell powder used as a substrate, it contained abundant protein and chitosan, and there were sufficient contents of nitrogen, phosphorus, and potassium, which are the three elements of fertilizer (Table 1). ).

Therefore, the RM, RCM, and RM_002 medium were sufficiently confirmed to be used as a mass growth medium of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain.

In addition, the growth of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain was confirmed for 7 days using an RCM medium rich in nutrients, and as a result, it showed the best growth rate on the 3rd day. The case was determined as a maximum of 3 days (Fig. 3).

Table 1 shows the results of the analysis of 9 major nutrients for the rice water (RM) medium, 2% rice flour (RM_002) medium and the active ingredient for crab skin powder.

Item Rice water 2% rice flour Crab shell powder Fat (g/100g) 0 0 - Sugars (g/100g) 0.32 0.23 - Cholesterol (mg/100g) 0 0 - Calories (kcal) 2.6 6 - Carbohydrate (g/100g) 0.5 1.24 - Trans fat (g/100g) 0 0 - Saturated fat (g/100g) 0 0 - Protein (g/100g) 0.15 0.26 - Sodium (mg/100g) 1.43 0.65 - Crude protein (%) 25.44 Calcium (mg/g) - - 132.43 Chitosan (mg/g) - - 43.02 Phosphorus (mg/g) - - 23.24 Total nitrogen (%) - - 4.15 Potassium (mg/g) - - 1.83

(Example 4) Enzyme activity of decomposing organic substances of a novel Bacillus velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain analysis

In order to confirm the activity of the organic substance degrading enzyme of the Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) isolated and identified in Example 1 above, starch degrading enzyme (amylase) and protease (protease) were prepared in the selection medium. ), lipolytic enzyme (lipase), and fibrinolytic enzyme (cellulase) as a result of confirming the enzyme activity against the new Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) confirmed that the organic substance degrading enzyme activity is excellent. (Fig. 4A to Fig. 4B).

(Experimental Example 1) Investigation of starch degrading enzyme activity

A plate medium was prepared in which 0.5% soluble starch, an inducer of amylase production, was added to the TSA medium. Bacillus Velezensis HY3479 of the present invention (Bacillus Velezensis HY3479, accession number: KCCM12751P) of the present invention on a plate medium is inoculated by two strains by a replacement culture method, and then the optimum temperature for enzyme expression of the strain of the present invention is 30 to 40°C, most preferably 37°C. After incubation for 20 to 30 hours, most preferably, for 24 hours, the medium was stained with Gran's iodine solution (iodine crystral 1.0 g, potassium iodine 0.2 g, Distilled water 30 ml) to confirm the starch degradation prevention (clear zone). (Figure 4A).

(Experimental Example 2) Proteolytic enzyme activity investigation

To investigate protease activity, 2% skim milk and 1.5% agar were sterilized and mixed to prepare a medium, and then Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) of the present invention. The strains were inoculated by two strains as in Experimental Example 1 by a replacement culture method, and cultured at 37°C for 24 hours at 37°C, which is the optimum temperature and time for the enzyme activity of the strain of the present invention, and then a clear zone was confirmed (FIG. 4B).

(Experimental Example 3) Lipolytic enzyme activity investigation

After preparing a NA agar medium containing 1% tributyrin and 3 to 5% Tween 80 for lipase activity, the Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain of the present invention was replaced. Two strains were inoculated as in Experimental Example 1 by the culture method, and cultured for 24 hours at 30°C, which is the optimum temperature and time for the enzyme activity of the strain of the present invention, and then a clear zone was confirmed (FIG. 4C).

(Experimental Example 4) Fibrinolytic enzyme activity investigation

After preparing a TSA medium containing 1% CMC for fibrinolytic enzyme activity, the Bacillus bellezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain of the present invention was replaced as in Experimental Example 1 by a culture method. After inoculating each strain and incubating for 72 hours at 30°C, which is the optimum temperature and time for the enzyme activity of the strain of the present invention, stained with 0.1% congo red for 30 minutes and washed with 1M NaCl, a clear zone was confirmed (Fig. 4D).

(Example 5) Plant disease control effect of new Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain

Various antifungal for each organic solvent fraction obtained by sequentially fractionating the culture solution of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain using the RCM medium selected under the above vegetable medium conditions. The activity was investigated (FIGS. 5, 6A and 6B).

(Experimental Example 5) Antifungal activity of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain against plant diseases

In order to investigate the antifungal activity of the Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain against major plant pathogens, the Agricultural Microbial Bank (KACC), pepper sclerotic disease (Sclerotinia sclerotiorum ), pepper anthrax ( Coletotrichum acutatum ), Apple tree blight ( Phytophthora cactorum ), cucumber ultimum (Phythium ultimum ), tomato gray fungus ( Botrytis cinerea ), red pepper rot (Rhizoctonia solani ), and ginseng root rot ( Cylindrocarpon destructans ) were pre-divided and used. The pre-sold mycelium was cultured at 24°C for 7 to 25 days in PDA medium. Afterwards, the hyphae disk was placed in the center of a disposable Petri dish dispensed with a PDA medium using a cork borer with a diameter of 5 mm, and the inhibitory effect on each plant pathogen was investigated through 3 repeated experiments after replacement culture (Table 2 and FIG. 5). As a result, strong antifungal activity was confirmed against various plant pathogens. The antifungal activity inhibitory effect of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain against plant pathogens is shown in Table 2 below.

Plant pathogen Scientific name Antifungal activity (mm ² ) Pepper scleroderma Sclerotinia sclerotiorum 17 ± 1.0 Chili pepper Rhizoctonia solani 20 ± 1.2 Tomato Gray Mold Disease Botrytis cinerea 13 ± 0.9 Apple Tree Plague Phytophthora cactorum 12 ± 1.2 Ginseng root rot Cylindrocarpon destructans 15 ± 0.5 Pepper anthrax Colletotrichum acutatum 15 ± 0.2 Cucumber jam Phythium ultimum 4 ± 2.0

As shown in the above results, antifungal activity was shown against all plant pathogens used in the experiment, and in particular, the best antifungal activity was exhibited against capsicum disease.

(Experimental Example 6) Antifungal activity by active ingredient of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain

Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain was cultured in TSB and separated sequentially using organic solvents hexane, dichloromethane, acetylacetate, and butanol. The organic solvent was removed by separating into an aqueous layer to confirm antifungal activity against ginseng root rot pathogens and cucumber jaalok pathogens under the same conditions as described above (FIGS. 6A and 6BA, FIG. 6A and FIG. 6BB).

As a result, the active ingredients showing the plant disease control effect of the Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain of the present invention are ingredients having polar properties consisting of an etiacetate layer, a butanol layer, and an aqueous layer. Is expected. According to the previous report by Lee et al. (Journal of the Korean Mycology Society, 2016), Bacillus velezensis CC112 strain has no inhibitory effect on ginseng jalococcus, whereas Bacillus velezensis HY3479 (Bacillus Velezensis HY3479, entrusted with the present invention) No.: KCCM12751P) was confirmed to have excellent activity against the Cucumber Jaloc pathogen (Figs. 6A and 6BB).

(Example 6) Pot test and field test of Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain

Bacillus velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) culture medium grown in RCM medium, which is a vegetable medium, was examined for plant growth and the yield of peppers (FIGS. 7 and 8 ).

(Experimental Example 7) Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, Accession No.: KCCM12751P) Strain Promoting Growth of Oat Sprouts and Inhibitory Effect of Homeland Fungi

In order to investigate the effect of the culture medium of Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) grown in RCM medium on the growth of oats, commercially available soil was laid on a pot, and 5 grains of oat seeds were seeded on the soil.

On the 7th day after sowing, suppression of fungi and oat sprout growth on the top soil was obtained by about 6 cm growth compared to the control group not treated with Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain culture solution (FIG. 7).

(Experimental Example 8) Plant Disease Control and Growth Promotion Effect of Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, Accession No.: KCCM12751P) Strain Against Outdoor Red Pepper

Red pepper was planted in the open field at Pohang Nodaji Village Co., Ltd. farm for Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) grown in RCM medium in large quantities, and an empirical test was conducted under the conditions shown in Table 3 below for 2 months. I did. As a result of conducting an empirical test for 2 months, the growth and yield of peppers sprayed with strains HY-003 and HY-007 (Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P)) increased significantly. The pepper yield was significantly higher than that of the positive control group (FIG. 8).

Target crop Way Remark Spray method Foliar spray
-2019.04.18 ~ 2019.06.04. : Once a week, 500 times diluted solution 250 mL
2019.06.12. ~ 2019.06.18. : Once a week, 1000 times diluted solution 400 mL Spray method Test Methods 2019.04.19. : Pepper seedling set meal
2019.05.02. : Red pepper net cleanup
2019.05.21. : Remove early open pepper
2019.06.18. : End of experiment term 2019.04.19 ~ 2019.06.18. (2 months) group Negative control (n=5) Rice water undiluted solution Positive control (n=5) P company best product Test group (n=5) HY-003 Test group (n=5) HY-007 strain
Named Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) and deposited the strain

The novel Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) strain of the present invention has an organic substance degrading enzyme activity and excellent antifungal activity against various plant pathogens, and also a low-cost plant medium (RCM, RM). It has the advantage of reducing the price burden on farmers as it can cope with the existing expensive microbial agents.

Therefore, Bacillus Velezensis HY3479 (Accession No.: KCCM12751P) is a low-cost, eco-friendly microbial agent that promotes plant growth with a biological control effect against various plant pathogens, and has sufficient industrial applicability.

Korea Seed Association KCCM12751P 20200618

Claims (10)

delete New microorganism Bacillus Velezensis HY3479 (Bacillus Velezensis HY3479, accession number: KCCM12751P) strain for promoting plant growth and controlling plant diseases was cultivated in a rice water medium containing crab skin powder, obtained by culturing the cells, culture medium, culture filtrate, or a combination thereof Containing as an active ingredient,
The plant diseases are, Sclerotinia sclerotiorum , Cretotrichum acutatum , Phytophthora cactorum , Phythium ultimum , Botrytis cinerea ), Rhizoctonia solani (Rhizoctonia solani), and Cylindrocarpon destructans (Cylindrocarpon destructans) a plant protective agent, characterized in that derived from at least one pathogen selected from the group consisting of. delete The method of claim 2,
A plant protective agent comprising an ethyl acetate layer, a butanol layer, an aqueous layer or a combination thereof as an active ingredient as a fraction obtained from the culture solution or culture filtrate of the Bacillus bellegensis HY3479 strain, and exhibits a plant growth promoting effect. delete Bacillus velezensis HY3479 (Accession No.: KCCM12751P) strain was cultivated in a rice-water medium containing crab skin powder, and obtained by culturing the cells, culture medium, culture filtrate, or a plant protection agent containing a combination of these as an active ingredient. Or, using the foliar spraying method, it exhibits the effect of controlling plant diseases,
As a fraction obtained from the culture medium or culture filtrate, a plant protective agent containing an ethyl acetate layer, a butanol layer, an aqueous layer or a combination thereof as an active ingredient exhibits a plant growth promoting effect by using a seed immersion or foliar spraying method,
The plant diseases are, Sclerotinia sclerotiorum , Cretotrichum acutatum , Phytophthora cactorum , Phythium ultimum , Botrytis cinerea ), Rhizoctonia solani (Rhizoctonia solani), and Cylindrocarpon destructans (Cylindrocarpon destructans) control method using a plant protection agent, characterized in that derived from one or more pathogens selected from the group consisting of. delete The method of claim 6,
Control method using a plant protection agent, characterized in that treating the plant protection agent at a concentration of 10 ⁵ to 10 ^{9 cfu/mL.} The method of claim 6,
The plant disease control effect is a control method using a plant protection agent, characterized in that exhibited in a plant selected from the group consisting of peppers, beans, barley, oats, cabbages, strawberries, cucumbers, ginseng and chives. delete

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