KR100312731B1 - Use of a novel Bacillus subtilis strain to protect the browning of Ginseng - Google Patents

Abstract

The present invention relates to a novel Bacillus subtilis strain B-4228 and a composition for preventing redness of a plant containing the same, and specifically, the strain of the present invention is a penicillium perfurosenum ( Penicillium) which is a causative agent of ginseng redness. It inhibits the red pigment production of purpurogenum ) and inhibits the growth of Trichoderma viride , another causative organism. Therefore, the novel Bacillus subtilis strain B-4228 provided by the present invention can be used as an environmentally friendly pesticide capable of preventing ginseng redness.

Description

Use of a novel Bacillus subtilis strain to protect the browning of Ginseng}

The present invention relates to Bacillus subtilis strain B-4228, which inhibits red pigment production of Penicillium purpurogenum , and a composition for preventing redness of a plant containing the same. More specifically, the present invention provides a novel Bacillus subtilis strain B-4228, which inhibits the production of red pigment of penicillium perfurosenium, which is a causative agent of red ginseng, and the growth of Trichoderma viride . It relates to a composition for preventing redness of plants.

Korean ginseng is an important natural resource unique to our country. It is a representative herbal medicine of Korea, which has been widely used for thousands of years as an oriental medicine or a medicine for oriental medicine as well as for human health. As Western medicine has shown limitations in overcoming the side effects of synthetic drugs and intractable diseases, the value of herbal medicines including ginseng is emerging. Ginseng prevents cancer, inhibits aging, protects the liver, strengthens fatigue and improves brain function, and recently has been reported to prevent the damage of environmental hormones. Ginseng, the chief of all herbal medicines, is a plant that grows in an extremely limited environment. It has a longer growth period than other crops and its growing method is special and difficult, and has been produced only in selected countries around the world.

Korea was a ginseng seed country by producing Korean ginseng with excellent medicinal effects due to its climate and climate as a natural growth and planting area of ginseng. However, many countries are paying attention to the ideal condition that ginseng has as a health food or health drug, and not only existing ginseng producing countries but also non-producing countries are actively promoting policies to encourage the research and production of ginseng. Ginseng growers include Korea, China, Japan, the United States, Canada, and parts of Europe. Most of the countries that consume ginseng are Confucian culture, including Korea, China, Japan, Taiwan, and Southeast Asia. Investigate the ginseng cultivation area, production volume, and consumption market size of each country in the world to collect statistical data accurately based on the characteristics of ginseng distribution structure, variety of product types, and security measures of recent data. Although Korea's ginseng production statistics show that Korea's ginseng production was 46% of the world's ginseng production by the late 1980s, Korea's share was around 39% in the 1990s. While the production growth rate was 0.5%, which is less than the world average production growth rate, China, which occupied 43% of the world production in the 80s, increased to more than 50% in the 1990s, and produced an annual average of 7.2% or more. The rate of increase has been chasing Korea, the ginseng species dominant country. (See table 1 )

World Ginseng Production Status (Based on Fresh Ginseng) Unit: 1,000 tons country 1987 (%) 1990 (%) 1992 (%) 1993 (%) Annual average increase and decrease (%) Korea 14,424 (46) 13,889 (42) 13,508 (38) 14,874 (39) 0.5 China 13,396 (43) 15,536 (47) 17,900 (51) 19,200 (50) 7.2 Japan 534 (2) 412 (1) 326 (1) 303 (1) -7.2 United States of America 2,356 (8) 2,367 (8) 2,379 (7) 2,803 (7) 3.2 Canada 456 (1) 744 (2) 904 (3) 972 (3) 18.9 system 31,166 (100) 32,948 (100) 35,107 (100) 38,152 (100) 3.7

1) Figures in () show the percentage of world production by country.

2) Source: Korea Tobacco & Ginseng Corporation

Korean ginseng is divided into red ginseng and white ginseng. White ginseng is mainly grown for 4 years. The types of products produced are processed and distributed in the form of fresh ginseng, white ginseng, taeguk ginseng, and dried ginseng. It is a forge that produces 6 years old ginseng and purchases almost all of them in the form of ginseng. Regionally, the border between Chungbuk and the red ginseng is predominant in the north and white ginseng in the south.

Ginseng production increased from 1,877 thousand tons in 1970 to 4,768 thousand tons in 1980 and 15,132 thousand tons in 1991, but decreased to 11,971,000 tons in 1995. The trend of ginseng production increased by 9.8% per year in the 1970s and 11.1% per year after 1980, indicating that ginseng production increased significantly since the mid-80s, but remained constant in the 1990s.

Many researchers have been concentrating on the development of scientific ginseng cultivation technology, including the establishment of ginseng production base, the improvement and management of seed, improvement of agricultural structure, improvement of cultivation methods and promotion of mechanization. In particular, ginseng is cultivated under sunscreen structure, unlike other crops, and according to environmental factors such as planting location, rainfall, temperature, wind intensity, soil fertility, moisture content, breathability, and acidity, The mutations tend to be very severe. Therefore, Korean ginseng has not yet been differentiated, but mixed native species are cultivated, and the cultivation method is inferior to other major crops due to the late introduction of scientific modern technology despite the long tradition. In order to produce high quality ginseng and ginseng products, the quality of raw ginseng should be good. Despite the improvement of seed growth, management, and cultivation methods of ginseng, and the improvement of the method of growing and managing the ginseng, the root skin of ginseng is reddish brown. There is a case of degrading the quality.

Red ginseng, which is said to have sulfur or sulfur by ginseng cultivators, begins with yellow spots on the epidermis of the roots and spreads to the entire surface. The growth of the roots is slowed down, and it gets worse as the old root grows, causing corruption. When red ginseng is cut, the central or outer layer is milky white, but the outer layer is yellowish brown, and even a part of the cortex is yellow. The reddish cortex is three times thicker than healthy ginseng and destroys the outer cells of the cortex cell layer. It is also known that yellowish cell contents gather near the cell membrane and the cell membrane is also colored yellow-yellowish brown.

Analysis of ginseng skin is mainly composed of fiber (30.9%) and crude lignin (23.7%), and other proteins, sugars and pectin, etc. The red skin is low in sugar content and crude greenin (Schuberin) component many. As a result of extract analysis of red urine from red ginseng ginseng, polyphenol, a precursor of lignin, was found to be combined with fat in large amounts. Therefore, epidermal redness is known to inactivate the cortical tissues by releasing large amounts of phenolic substances in the process of abnormal corkization of the root cortex, combined with fatty substances, or polymerized into polymers, and oxidized and red. Ginseng that has advanced root redness decreases the absorption of moisture and decreases the respiration of roots, and decreases the absorption of potassium and manganese among inorganic components, while increasing the absorption of iron, soda, and phosphoric acid, and the K / Na ratio of ginseng. Nutrient imbalance of the Fe / Mn ratio. In particular, a large amount of iron is detected in the reddish hull, which may be due to iron being directly involved in red change or deposited in a large amount of polyphenols.

Thus, physiological disturbance caused by redness decreases the growth of the roots and causes redness at the time of two years, which causes about 50% of productivity loss at four years of age. There is a lot of coarse, yellowish brown or rough skin, the quality is very poor. The ginseng root's outer sheath, like other plants, breathes, helps to absorb the necessary nutrients, protects the inside of the roots, increases in content as the root grows, and corks grow as growth progresses. However, due to internal and external factors, physiological functions are disturbed, corkization is promoted, or phenolic substances cause abnormal polymerization, which causes redness. Therefore, all factors that disturb the physiological metabolism of roots may promote red change of roots.

Common causes of redness in ginsengpo include:

First, excessive soil moisture impairs soil air permeability, inhibits root respiration due to lack of oxygen, inhibits metabolic activity of roots, and disturbs physiological functions by accumulating a large amount of reducing substances.

Secondly, the application of immature organic fertilizers causes oxygen in the soil air during the maturation of organic matters, causing respiratory failure of the roots and generating toxic substances such as ammonia and organic acids, which can cause redness in the roots.

Third, redness may occur due to mechanical damage of ginseng. When the root tissues are damaged by splitting of the body, dropping of the root muscles and insects, animals, etc., polyphenols may cause oxidative condensation in the process of forming a protective film on the site.

Fourth, redness may occur due to physical disorders. Ginseng roots have lower moisture tension than other crops, so the metabolism of roots is easily disturbed under conditions of high osmotic pressure outside the roots, i.e., soil drying and defoliation, resulting in effusion of phenolic substances.

Fifth, it may occur due to the chemical factors of the soil. If a large amount of metallic substances such as iron is present in the soil, the phenolic substance of the skin is directly oxidized and condensed, resulting in redness.

Sixth, nutrition may occur as a physiological factor. It is known that red ginseng may occur due to internal physiological disturbances when potassium is deficient and excessive iron is absorbed during the growth of ginseng.

Care should be taken from the site selection in order to reduce root redness of ginseng.In case of poor conditions even after the construction of sunlight and rain cover, redness may occur within about a week. The fields where water comes out or the water accumulates in the wetland should be excluded from the planned site.In addition, where fertilizer residues are high by using chemical fertilizers in all crops, it may be excluded if it is difficult to check and improve the salt concentration or test the phosphoric acid content. In case of selecting a flat field as ginsengpo, it is recommended to install sufficient drainage on the side of Sampo to prevent excessive rainwater from accumulating in the furrow, and to prevent the inflow of surface water from other fields, even if the site is well selected and the drainage management is carried out thoroughly. Development of roots Bad, elongated roots also cause redness due to poor breathing, so you need to grind a lot of fresh leaves such as leaves and soften the soil, and set the height of the toad as high as 30cm, or use abundant organic fertilizer or livestock meal or use it late. In this case, it is recommended to avoid the application of rich organic fertilizers and livestock manure, as the roots are changed by reducing the soil as it causes salt disorders or progresses in housing. Active active carbon (150-300 kg per 10a) was used to control the redness by adsorbing harmful substances. (Choi Gwang-tae's latest Korean ginseng cultivation, Korea Ginseng and Tobacco Research Institute, Cheonil Print Company 113-116, 1996).

Red ginseng has a red or reddish brown color, and the surface of the ginseng is red or reddish brown. The surface is rough and poor development of roots and roots reduces productivity. When red ginseng is manufactured, red ginseng has a brownish color and the product becomes crude. Will be done. Red ginseng products are also subject to banned pesticide regulations in foreign countries, so the use of prohibited chemical synthetic pesticides should not be used. However, in order to prevent red ginseng, many researchers have tried to control with various cultivation methods and chemical synthetic pesticides, but the low redness control value and some farmers use pesticides indiscriminately to contaminate water quality and soil.

The present inventors are antagonistic microorganisms that can prevent red ginseng from microbial resources in order to develop pesticides that can prevent redness, to develop pesticides with low toxicity, selective drug efficacy, excellent sustainability and pharmacokinetic properties, and environmentally friendly. Was explored. As a result, the present inventors not only searched and identified the antagonistic microorganisms that inhibit the growth of the strain after separating the strains causing the redness from the ginseng in progress, as well as the actual ginseng farmland using the microorganisms as a pesticide for preventing redness The present invention was completed by treatment with and confirming the effect.

It is an object of the present invention to provide an environmentally friendly pesticide that is excellent in redness prevention against ginseng.

It is also an object of the present invention to provide a microorganism that can be used in a composition capable of preventing redness, such as ginseng.

It is also an object of the present invention to provide a use of the microorganisms to inhibit the growth of the causative strain of ginseng redness or the production of red pigment.

1 is a culture photograph of the erythrocyte producing strain,

Left plate: incubation of Penicillium purpurogenum (top spot) and Trichoderma viride (bottom spot) at the same time;

Right plate: Streaking Bacillus subtilis B-4228, the active microbial strain of the present invention, between two spots of the left plate,

2 is a gas chromatogram showing the intracellular fatty acid content of Bacillus subtilis B-4228,

3 is an electron micrograph showing the surface of the active microbial strain of the present invention,

Figure 4 is an electron micrograph showing the flagella of the active microbial strain of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides a novel Bacillus subtilis strain B-4228 (Accession Number: KCTC 0657BP).

Characteristic of the strain is that it has the activity of inhibiting the growth of red pigment production and the growth of Trichoderma viride , Penicillium purpurogenum , the causative agent of ginseng redness. The present inventors have isolated the strains causing redness from red ginseng in progress, and confirmed that these microorganisms are penicillium perfurosenium and trichoderma viridi. When the microorganisms were cultured alone, the production amount of red pigment of penicillium perfurosenium was small, but when they were cultured together, the penicillium perfurosenium was found to produce many red pigments (see FIG. 1 ). That is, the inventors first discovered that the red pigment production of penicillin perfurosenum causes ginseng redness, and that the increase in the red pigment production is possible only when the trichoderma viridi is present together. Therefore, the present inventors have searched for a microorganism capable of inhibiting the proliferation of Trichoderma viridi, to develop a microorganism for preventing ginseng redness.

Bacillus subtilis strain B-4228, the strain of the present invention, was isolated in the process of searching for a microorganism having an effect of inhibiting the growth of trichoderma viridi, a causative agent of ginseng reddening, specifically from a soil sample of Geumsan, Chungcheongnam-do. Separated.

The sequence of the gene encoding the 16S rRNA of the strain provided by the present invention (rDNA) is set forth in SEQ ID NO: 1 . Compared with the rDNA sequence of the known strain, the rDNA sequence described as SEQ ID NO: 1 shows 99% similarity with the known Bacillus subtilis strain AB017588, and 99% identity with another known Bacillus subtilis strain AB017589. Shows.

When comparing the characteristics of the fatty acid in the cells of the strain provided by the present invention with known microorganisms, it is most similar to Bacillus lentimorbus , but the agreement is low at 58.4%. On the other hand, the agreement with Bacillus subtilis was lower, 49.9%.

The size of the strain provided by the present invention is 1.5 to 1.7 μm in length and 0.6 to 0.8 μm in width, which is smaller than 1.2 to 2.5 μm, which is the size of a general Bacillus subtilis (see FIG. 3 ). About five times its size (see FIG. 4 ).

The inventor classified the new strain into Bacillus subtilis based on the comparison data of 16S rDNA sequences of microorganisms most used to identify the location of the new strain. Based on the characteristics of the strain of the present invention was named Bacillus subtilis B-4228, deposited on September 21, 1999 in the Institute of Biotechnology Gene Source Center (Accession Number: KCTC 0657BP).

The present invention also provides a method for preventing redness of ginseng using Bacillus subtilis strain B-4228.

The method consists of applying Bacillus subtilis strain B-4228 or a composition comprising said strain to ginseng arable land or ginseng itself. As the method of applying the strain or the composition thereof, spreading, spraying, soaking directly with the culture medium of the strain, or by spreading the strain culture medium by adsorbing the diatomaceous earth, perlite, or management of arable land In this case, a method of mixing the strain culture solution or the adsorbed strain during the soil change can be used. In addition, known pesticide administration methods, microbial application methods and the like can be used.

Penicillium perfurosenium and Trichoderma viridi have been identified as the causative agents of ginseng redness. When these microorganisms are cultured alone, they produce less red pigment, but when they are cultured together, they are rich in penicillium perfurosenium. To produce (see FIG. 1 ). When Bacillus subtilis strain B-4228 is incubated with Trichoderma viridi, the growth of Trichoderma viridi strain is inhibited. In addition, when Trichoderma viridi, Penicillium perfurosenium and Bacillus subtilis strain B-4228 are incubated simultaneously, not only growth of Trichoderma virididi but also red pigment production by penicillium perfurosenium Is inhibited.

In the embodiment of the present invention, by treating the strain culture solution in various ginseng arable land near the ginseng root, the degree of improvement of red ginseng for the treated and untreated strains were investigated. As a result, it was confirmed that the ginseng red yeast ratio was reduced in all of the treated arable land compared to the untreated control. Therefore, it was confirmed that the ginseng redness prevention method of the present invention can be usefully used in actual ginseng farmland.

In addition, the present invention provides a composition for preventing vegetative alteration containing Bacillus subtilis strain B-4228.

The composition comprises Bacillus subtilis strain B-4228, a novel strain of the present invention, as an active ingredient. The strain may be supplied as contained in the composition for preventing redness of the present invention, but may be stored separately for long term storage and then mixed and used immediately before use. When the strain is supplied separately for long-term storage, the strain may be included in a 10% glycerol solution and stored at a low temperature of about -80 ° C or suspended at a high concentration in sterilized 10% skim milk or the like and freeze-dried.

Other components included in the composition may include excipients, stabilizers, other additives known in the conventional pesticide manufacturing field, in particular in the field of biological pesticide production containing microorganisms, or may be combined with diatomaceous earth, perlite, ocher, and the like.

The composition may be used to prevent redness of the ginseng by treating such as ginseng being grown or harvested ginseng.

When spraying on the ginseng being cultivated, that is, ginseng arable land, it is preferable to spray concentrated on the root tissue of the ginseng or the soil close to it, and preferably sprayed 2 to 4 times at a monthly interval from April. The effective amount of the microorganisms contained in the composition for preventing redness of the present invention is 1 × 10 ⁷ or more per cultivated land area (m 2).

On the other hand, in the case of applying the composition for preventing redness of the present invention to ginseng after harvesting, it may be directly sprayed on the root tissue of ginseng, or in a solution diluted to a concentration of 1 × 10 ⁶ ~ 1 × 10 ⁹ microorganisms per milliliter 24 It can also be immersed for 144 hours.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Isolation of Causative Bacteria and Observation of Microbial Ecology of Ginseng Redness Phenomenon

Strains causing redness were isolated from ginseng in progress. Specifically, PDA medium (potato 30%, glucose 2%, agar 1.5%, pH 5.1, 121 ° C., 15%) prepared in advance by peeling off the area where the redness of ginseng in progress of red ginseng purchased from Chungnam-negative fat is in progress with a sterile knife. Minute sterilization) onto the plate and incubated strain-related strains for about 72 hours at 28 ℃.

As a result of taxonomic studies of the isolated causative organisms, these microorganisms were identified as Penicillium purpurogenum and Trichoderma viride . Taxonomic studies of penicillium perfurosenum are described in Samson RA et al; Introduction of food-borne fungal, Netherlands, Centraalbureau voor Schimmelcultures pp.573-575, 1995, and the taxonomic studies of trichoderma viridi were described by Christian P. and Gary EH, Trichoderma and Gliocladium, Basic biology, taxonomy and genetics, Vol. 1, pp. 3-30, 1998.

On the other hand, when the two microorganisms are cultured alone in MEA medium (malt extract 2%, peptone 0.5%, agar extract 1.5%, distilled water 1L), respectively, the production of red pigment is small, but when in close proximity, penicillium perfurosenum It was confirmed that a large amount of this red pigment was produced (see the left plate of FIG. 1 ).

Example 2 Screening of Antagonist Microorganisms That Inhibits Red Ginseng

The causative organisms isolated in the above examples were used to search for antagonistic microorganisms that inhibit the growth of microorganisms related to red pigment production so that the causative organism of ginseng redness cannot produce red pigment.

Specifically, antagonistic microorganisms with red pigment production inhibitory activity were examined in soil samples of Geumsan, Chungcheongnam-do. Dilute the soil sample in distilled water, while separating microorganisms (yield extract 0.1%, polypeptone 0.5%, K ₂ HPO ₄ 0.05%, KH ₂ PO ₄ 0.05%, MgSO ₄ 7H ₂ O 0.02%, agar 1.5) %, pH 7.2), sterilized and cooled to 50 ℃, and then added colistin to 5 mg / l in distilled water as an additional antibiotic, and Nalidixic acid to 10 mg / l in methanol. 1 ml per 1 liter of sterilized medium was prepared. Soil samples diluted in distilled water were added to the prepared medium and cultured at 28 ° C. to isolate the bacteria. In search of active microorganisms, antagonistic microorganisms having inhibitory activity in the growth of Trichoderma viridi strains were selected while culturing Trichoderma viridi strains in PDA medium.

Example 3 Identification of Antagonistic Microorganisms Discovered

In order to identify the location of the antagonistic microorganisms searched, 16S rRNAs of antagonistic microorganisms were isolated and DNA sequences ( SEQ ID NO: 1 ) were determined. GenBank's BLAST search program was used to analyze gene similarity with known strains. It was. As a result, the rDNA of Bacillus subtilis AB017588 had 495th, 628th, and 867th bases in the base sequence of SEQ ID NO: 1 with adenine, and the 1208th base sequence was guanine, and 1439 bases were identical among 1443 bases. (99% consistency). On the other hand, Bacillus subtilis rDNA of Antilles AB017589 is 1439 bases in the 495th, 867th nucleotide in the nucleotide sequence of SEQ ID NO: 1 is the adenine and the 1415th base is adenine, it 1416th base is a thymine 1443 bases The sequence was identical with the nucleotide sequence of SEQ ID NO: 1 (99% identity).

Antagonistic microorganisms searched to identify the classification position by comparing the components and content of fatty acids inherently contained in each microorganism (Shin et al , J. Gen. Appl. Microbiol., 43, 317-324,1997) Intracellular fatty acids were extracted and quantitatively analyzed by gas chromatography (see Fig. 2 ). In Sherlock Version: 2.95, the components and ratios of constituent fatty acids of existing microorganisms in the database were compared and investigated. . As a result, the components and ratios of fatty acids that make up the isolated antagonist microorganisms were the most similar to those of Bacillus lentibus, but showed low concordance of 58.4%, and lower concordance with Bacillus subtilis (49.9%). Appeared.

External morphology of antagonistic microorganisms was observed by electron microscopy. As a result, the isolated antagonistic microorganisms were 0.6-0.8 μm × 1.5-1.7 μm in size, which was smaller than 1.2-2.5 μm, which is the size of the general Bacillus subtilis (see FIG. 3 ). About 5 times (see FIG. 4 ).

The present inventors classified active microorganisms into Bacillus subtilis based on the comparison data of 16S rRNA sequences of microorganisms which are most used to identify the positions of microorganisms, and the composition and ratio of fatty acids of active microorganisms, cell appearance, etc. Based on the characteristics of the antagonist microorganisms were named Bacillus subtilis B-4228 and deposited on September 21, 1999 at the Institute of Biotechnology Gene Source Center (Accession Number: KCTC0657BP).

Example 4 Preparation and Growth of Cell Concentrate of Bacillus Subtilis B-4228

Antagonistic microorganisms with inhibitory activity against ginseng reddening were 1: 1 ratio of culture medium and 20% glycerin at 28 ° C for 24 hours in LB medium (1% casein peptone, 0.5% yeast extract, 0.5% salt, after autoclaving). The mixture was mixed with 2 ml frozen vials and stored at -70 ° C and used for culturing active microorganisms.

To produce active microorganisms, the frozen Bacillus subtilis B-4228 vial was dissolved, and the active microbial production medium (1% soluble starch, 0.5% pharmacy 0.5%, glucose 0.2%, ammonium sulfate 0.1%, phosphoric acid) in a 1 liter Erlenmeyer flask. Potassium 0.1%, magnesium sulfate 7-fold crystalline water 0.05%, calcium chloride 0.1%, sodium chloride 0.3%, initial pH 7.0) was inoculated in 100ml of the medium was incubated at 28 ℃ for 48 hours at 200rpm speed.

Example 5 Inhibitory Effect of Tricoderma viridi Strains on Bacillus subtilis Strain B-4228 and Inhibition of Red Pigment Production by Penicillium Perfurosenum

In order to confirm that Bacillus subtilis strain B-4228 inhibited the growth of Trichoderma viridi strains, the growth inhibitory activity of the Trichoderma viridi strains was analyzed in PDA medium. Take Tricoderma viridi incubated in PDA medium for 28 hours at sterile white gold tooth and draw it to date. Incubate in PDA medium and take Bacillus subtilis strain B-4228 with sterile white gold tooth. Then, after 28 hours incubation for 72 hours, it was confirmed that the growth of Trichoderma viridida strain is inhibited.

In addition, in the same manner as above, Trichoderma viridi, Penicillium perfurosenium, and Bacillus subtilis strain B-4228 were simultaneously cultured. As a result, Bacillus subtilis strain B-4228 was grown with Trichoderma viridi. In addition, the red pigment production of penicillium perfurosenum was also inhibited (see right plate of FIG. 1 ).

Example 6 Inhibitory Effect of Bacillus Subtilis Strain B-4228 on Ginseng Redness (1)

In order to confirm the inhibitory activity of the active microorganisms in the ginseng cultivation area, it was tested in farmland in Chungcheongnam-do, Chungcheongbuk-do, and Gyeonggi-do. Determination of the redness improvement of ginseng for the treatment of active control against redness was determined by Kim Myung-su of Samwon Nongsan Co., Ltd., Jeungpyeong-eup, Chungnam.

Production medium of Example 4 in May 1999 in 200 of the five-year-old ginseng cultivated in Yeongsong-ri, Yeongpung-myeon, Pocheon-gun, Gyeonggi-do, Korea (size of 1.8 ginseng and 50 roots of cultivated ginseng). Diluted antagonistic microorganisms cultured with 25 times to adjust the number of microorganisms to 4 × 10 ⁷ per milliliter and then watered once at 5 liters per column, and redness improvement of ginseng for the treatment of antagonist antagonistic strains in August 1999 Was investigated. The calculation of the redness index is 4 at 31-100%, 3 at 31-60%, 2 at 16-30%, 1 at 6-15%, and 0 at 0-5% depending on the degree of redness. The degree of deficit was expressed quantitatively.

When the redness rate was calculated using this formula, the redness rate of the untreated group was 72.6% and that of the antagonist microorganisms was 54.2%. These results indicate that the ginseng grade improved moderately severe redness to moderate redness.

Example 7 Ginseng Redness Inhibitory Effect of Bacillus Subtilis Strain B-4228 (2)

Gyeonggi-do Yeoncheon County jangnammyeon sugar 1 Lee one antagonistic microorganism culture in ginseng cultivation in production medium of Example 4 in May 1999 to 170 spaces of the five year-old ginseng 3,000 spaces being cultivated diluted 25-fold the number of microorganisms 4 × 10 ⁷ per miririteo After the adjustment, the water was irrigated once with 5 liters per column, and the improvement of redness of ginseng was examined in August 1999. Calculation of redness improvement and redness index of ginseng for the treatment regimen active redness was calculated as in Example 6.

When the ratio was calculated, the percentage of untreated group was 82.0% and that of antagonist microorganism was 75.0%. This result was a slight improvement in severe redness in ginseng grade.

Example 8 Ginseng Redness Inhibitory Effect of Bacillus Subtilis Strain B-4228 (3)

Dilute the antagonistic microorganisms cultured with the production medium of Example 4 in 350 out of 4,000 bales of 4000 year-old ginseng in the Songwoo Ginseng Farmland, Pocheon-eup, Pocheon-gun, Gyeonggi-do, and adjust the number of microorganisms to 4 × 10 ⁷ per ml. After that, 5 liters of water per water were irrigated once, and the improvement of redness of ginseng was investigated in August 1999. Calculation of redness improvement and redness index of ginseng for the treatment regimen active redness was calculated as in Example 6.

When the redness rate was calculated, the redness rate of the untreated group was 82.0% and that of the antagonistic microorganisms was 75.0%. This result was a slight improvement in severe redness in ginseng grade.

Example 9 Inhibitory Effect of Bacillus Subtilis Strain B-4228 on Ginseng Redness (4)

Diluted antagonistic microorganisms cultured in the production medium of Example 4 in July 1999 in 80 of the 5000 four-year-old ginseng planting in Ginseng Farmland, Sangdeok-ri, Godeok-myeon, Godeok-myeon, Chungcheongnam-do, the number of microorganisms was 4 × 10 ⁷ per liter in advance. After adjustment, the water was irrigated once at 5 liters per column, and the improvement of redness of ginseng was examined in August 1999. The calculation of the redness index was calculated as in Example 6.

When the redness rate was calculated, the free redness was 59.3% and that of the antagonistic microorganisms was 47.3%. The results showed a slight improvement in severe redness when expressed by ginseng grade.

Example 10 Inhibitory Effect of Bacillus Subtilis Strain B-4228 on Ginseng Redness (5)

The antagonistic microorganisms cultured in the production medium of Example 4 were diluted 25 times in 170 out of 700 three-year-old ginseng cultivated at Ginseng Farmland, Songjeong-ri, Cheongwon-gun, Chungcheongbuk-do, and adjusted the number of microorganisms to 4 × 10 ⁷ per ml. In August 1999, the degree of redness improvement of ginseng was investigated in August 1999. The calculation of the redness index was calculated as in the above example.

When the redness rate was calculated, the redness rate of the untreated group was 19.5% and that of the antagonistic microorganisms was 10.1%. This result shows that less redness is improved to less redness by ginseng grade.

The novel Bacillus subtilis strain B-4228 provided by the present invention effectively inhibits the growth of Trichoderma virididi and penicillium perfurosenum, which is the causative agent of ginseng, and effectively inhibits the red pigment production of penicillium perfurosenum. It can be used as an eco-friendly ginseng anti-pesticide because it has no pollution and human toxicity.

Claims (9)

Penny room Solarium PARFUMER rose to more than Bacillus inhibit the production of a red pigment (Penicillium purpurogenum) subtilis (Bacillus subtilis) strain B-4228 (accession number: KCTC 0657BP). Composition for preventing plant redness containing the strain of claim 1. The composition for preventing redness of plants according to claim 2, which is applied to ginseng or the soil around it. 4. The method of claim 3, wherein the application consists of spraying concentrated ginseng roots or adjacent soils, wherein the effective amount of microorganisms is adjusted to 1 × 10 ⁷ or more microorganisms per cultivated area (m 2). A composition for preventing redness of a plant. The method according to claim 3, wherein the application is applied directly to the root of ginseng, diluted with a microbial concentration of 1 × 10 ⁶ to 1 × 10 ⁹ per ml, or 1 × 10 ⁶ to 1 × 10 ⁹ per ml. The composition for preventing redness of plants, characterized in that consisting of immersing ginseng in a solution diluted to a microbial concentration for 24 to 144 hours. Method of preventing redness of ginseng using the strain of claim 1. 7. The method of claim 6, wherein the strain of claim 1 inhibits the growth of Trichoderma viride . The method of claim 6, wherein the strain of claim 1 inhibits red pigment production of Penicillium purpurogenum . The method for preventing redness of ginseng according to claim 6, comprising applying Bacillus subtilis strain B-4228 or a composition comprising said strain to ginseng farmland or ginseng itself.