JP3140430B2 - Bacillus microorganisms and their uses - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a Bacillus cereus KI2N strain (FERMP-17).
147), compositions containing it, and uses thereof. More specifically, the present invention provides, in addition to the microorganism itself,
Fermentation promoter, fertilizer additive, soil conditioner and phytopathogenic fungal growth inhibitor containing the same, and fermentation promotion using such fermentation promoter, fertilizer additive, soil conditioner and phytopathogenic fungus growth inhibitor The present invention relates to a method, a plant cultivation method, a soil improvement method, and a phytopathogenic fungal growth suppression method.

[0002]

2. Description of the Related Art Modern agriculture always cultivates a single crop at a high density. As a result, specific components of the soil are consumed and the components are biased, and the components required for plants are insufficient, and phytopathogenic microorganisms, especially phytopathogenic fungi, accumulate in the soil. It has become.

[0003] Therefore, chemical fertilizers that have been heavily used to replenish only specific components have promoted the continuation of farming without supplementing other various trace components, and as a result, have greatly reduced the soil fertility. It also cut off the flow of organic matter that was originally circulating, and organic waste such as animal dung lost its place to go.

At the same time, chemical pesticides that have been widely used have a wide influence on the growth of microorganisms and insects that are effective for the growth of plants, and the ecosystems surrounding agriculture lose their balance and become very delicate. Was. Agriculture has also compromised the health of farmers, or pesticide residues have harmed the health of consumers.

[0005] In recent years, the importance of organic agriculture has been proposed as a countermeasure against these problems. The use of organic fertilizers will help maintain soil fertility, and farming itself will help create a recycling-oriented society. There is no negative impact on the health of farmers and consumers.

[0006] In recent years, the use of microorganisms has been studied, and agricultural and horticultural germicides using microorganisms have been proposed. However, many germicidal effects themselves are still insufficient.
In addition, in the soil, there are many disadvantages such as the fact that the fungus itself is killed or the efficacy is suddenly lost due to lack of sustainability, and the fact is that very few are satisfactory. Furthermore, satisfactory microorganisms which have not only antibacterial properties but also an organic matter fermentation promoting action, a plant growth promoting action and a soil bactericidal action and have a sustained action have not yet been obtained.

[0007]

SUMMARY OF THE INVENTION The present invention has been made in order to meet the demands of farmers and consumers for such organic agriculture. One microorganism has not only one action but also two or more actions. The purpose of the present invention is to cultivate new microorganisms that have these effects and maintain these effects.

[0008]

SUMMARY OF THE INVENTION The present invention has been made to achieve the above-mentioned object which is extremely difficult to solve.
After extensive screening of various microorganisms, Bacillus microorganisms promote the fermentation of organic matter by sensitizing them to organic matter, promote the growth of plants by being contained in fertilizer, and apply it to soil. It has been found for the first time that there are microorganisms that improve soil and control phytopathogenic fungi in soil or on plant surfaces by applying to soil or plants, and these effects are persistent. As a result of further intensive research, a new microorganism belonging to the genus Bacillus, Bacillus cereus KI2N strain (FERM P-17147), which succeeded in the above purpose, was successfully isolated.

[0009] That is, the present invention provides a Bacillus cereus KI2N strain having both the property of promoting the fermentation of organic matter and the property of inhibiting the growth of fungi. The present invention also provides a composition containing the above-mentioned microorganisms, which has various actions and has a long-lasting action, and has a wide application range. Effective continuous action fermentation promoter composition, continuous action fertilizer or fertilizer additive composition having a plant growth promoting effect, etc.
It is intended to provide a sustained-acting soil improver composition for sterilizing soil contaminated with harmful bacteria to improve the soil and the like, and further provide a sustained-acting phytopathogenic fungicidal or growth inhibitor composition.

[0010] The microorganism according to the present invention is Bacillus cereus KI2N strain and has the following properties. Cell swelling-NaCl 7% + Anaerobic growth + Starch degradation + VP-Egg-yolk-Indole-Growth at 60 ° C-Gram staining + Spore position Center and semi-end position Lusterless Colony surface Wrinkles Cell size 1.2x 3-3.5 μm DHL-MacConkey-mannitol medium-normal agar medium + catalase + urea degradation-oxidase + hemolytic + hydrogen sulfide-nitrate reduction + ONPG-acid production from carbohydrate galactose-D-mannose-melibiose-xylose-xylose L-arabinose-glucose + mannitol-rhamnose-glycerol-esculin +

From the above properties, this isolate is recognized to belong to Bacillus cereus, and has the further characteristic of promoting the fermentation of organic matter, promoting the growth of plants, and inhibiting the growth of fungi. Therefore, this isolate was recognized as a new strain, and was identified as Bacillus cereus KI2.
N, and on January 14, 1999, FERM P-17147 was registered with the Institute of Biotechnology and Industrial Technology of the Ministry of International Trade and Industry of the Ministry of International Trade and Industry.
(Hereinafter sometimes simply referred to as KI2N strain).

The KI2N strain shows good growth on both normal agar medium and 10% sheep blood-supplemented blood agar medium. In each medium, the colonies are R-type, gray-white, not glossy, and have a wrinkled surface.

To prepare the KI2N strain, it is desirable to perform pure culture using a solid medium or liquid medium. A normal agar medium can be used as a solid medium, and a meat extract medium can be used as a liquid medium. It is preferable to culture at about 37 ° C. under aerobic conditions. When it is necessary to consider the stability in formulation and distribution, it is preferable that the cells be in a spore state. Therefore, the culture time is set to a long time of about 48 hours, or a high-temperature environment of about 80 ° C. It is preferable to promote spore formation. In addition, it is possible to culture the soybean meal, food residue, livestock dung, etc. as a base material. In this case, a culture base with few germs is required in advance so that the KI2N strain grows superiorly to other microorganisms. Alternatively, it is necessary to use a sterilized culture substrate.

The KI2N strain used in the present invention includes, in addition to the isolated strain itself, its suspension or culture solution, culture,
Alternatively, it broadly includes processed products thereof (e.g., concentrates obtained by centrifugation, paste-like products, dried products obtained by freeze-drying, etc., and diluted products).

The KI2N strain-containing composition according to the present invention may use only the above-mentioned KI2N strain and / or a processed product thereof, or may further comprise vermiculite, kaolin,
Nontronite, beidellite, montmorillonite and other clay minerals; perlite; charcoal, burnt old tires, burned waste paper and other carbides; beer lees, sake lees, soy sauce lees, shochu lees, starch lees, bagasse, okara and other food production lees; Wood chips, pulp chips, sawdust and other forest waste; rice straw, wheat straw, buckwheat, fir and other agricultural waste;
Livestock waste; zeolite; peat moss and other carriers;
It may be carried or mixed on a substrate or the like.

Since the KI2N strain has an action of promoting the fermentation of organic substances (in other words, an action of decomposing organic substances), the above-described composition containing the KI2N strain can be used as a fermentation promoter composition.

When the fermentation accelerator composition is sensitized to an organic substance, the amount of the KI2N strain is preferably at least 10 ⁵ CFU per gram of the organic substance. However, there is no particular problem since the fermentation starts after waiting for a while even for a small amount. Fermentation accelerator composition is not only sprayed on the surface of organic matter,
Stir and mix well. Although the temperature rises with the promotion of fermentation, the KI2N strain proliferates and metabolizes more actively in an aerobic state than in an anaerobic state. Therefore, air containing oxygen is periodically removed by a method such as stirring or aeration. Preferably it is supplied. When the fermented product produced by this method is used as compost or other organic fertilizers, as organic substances, sludge, sludge dewatered cake, livestock dung, livestock urine,
Those that do not contain harmful substances such as human excreta, sawdust, beer lees, food residues, okara, and the above-mentioned carriers and base materials, or a mixture thereof are selected.

Agricultural materials, compost and other organic fertilizers produced by sensitizing the fermentation accelerator composition to organic matter can be used as such or mixed with other organic fertilizers.
It can also be applied by adding fertilizer additives. Both solid organic fertilizers, liquid and pasty organic fertilizers can be applied in accordance with conventional methods.

Since the KI2N strain has an action of promoting the growth of plants, the composition containing the KI2N strain can be used as it is as a fertilizer composition and / or a fertilizer additive composition. In the latter case, it may be used in addition to compost or other organic fertilizers as well as chemical fertilizers, or may be used in addition to the above-mentioned organic substances.

Furthermore, since KI2 strain N has an action of suppressing or killing the growth of phytopathogenic fungi and an antifungal action, the above-described composition containing KI2N strain is used as a soil conditioner composition and an antifungal composition. It can be used as a (fungal growth inhibitor composition or fungal fungicidal composition).

When these compositions are used, the cells of the KI2N strain and / or a processed product thereof may be used alone, but a carrier, a surfactant, a dispersant, and the like may be used in accordance with a conventional method for preparing agricultural chemicals. Alternatively, it is more preferable to add an auxiliary agent or the like and formulate it into a form such as a powder, a wettable powder, an emulsion, a flowable, or a granule and use it in a conventional manner. Suitable carriers include, for example, clay, talc, bentonite, diatomaceous earth, white carbon, kaolin, vermiculite,
Solid carriers such as slaked lime, silica sand, ammonium sulfate, urea and the like can be mentioned. Examples of surfactants and dispersants include metal salts of alkyl benzene sulfonic acid, polyoxyethylene alkyl aryl ether, sodium alkyl sulfate, sodium alkyl naphthalene sulfonate, and dinaphthyl. Examples include sodium methanedisulfonate and sodium ligninsulfonate. Examples of the adjuvant include carboxymethylcellulose, polyethylene glycol, gum arabic, starch, lactose and the like.

As a method of applying the soil conditioner composition to the soil, it may be merely sprayed on the soil surface, but it is preferable that the soil conditioner composition is sufficiently incorporated so as to be mixed with the surface layer of the soil. This composition is intended to improve the soil by killing or inhibiting the growth of microorganisms such as phytopathogenic fungi in the soil, and when organic matter is present in the soil,
Organic substances existing in the soil are decomposed by the action of various strong and persistent enzymatic forces of the KI2N strain, and chemical or physical soil improvement (for example, formation of an aggregate structure) can be performed. It is. In addition, the fungus does not die even in soil, and its effects are maintained.

When the antifungal composition using the KI2N strain is applied to soil, it is preferable that the antifungal composition not only be sprayed on the surface of the soil, but also sufficiently incorporated so as to be mixed with the surface layer of the soil. Moreover, when spraying on the leaf stem of a plant, it is preferable to adjust the pathogenic fungus fungicide or growth inhibitor using a spreading agent so as to remain on the surface. When coating the seeds, the seeds are immersed in a culture solution in which the KI2N strain is cultured and then dried, or the seeds are dressed and coated with a binder.

When the composition containing the KI2N strain according to the present invention is used for various purposes, the amount used and the amount of dilution are appropriately selected according to the dosage form of the composition, the method of application, the place of application, the time of application, the purpose of application, and the like. But K / g of applied material such as soil and organic matter
The range of 10 ² to 10 ¹² CUF is suitable for the I2N spore, and foliage application may be performed according to this. In addition, in the case of compost production, for example, the KI2N strain itself grows after application, so that the desired purpose can be sufficiently achieved at first even with a small amount of addition. Hereinafter, the present invention will be further described with reference to Examples, but the present invention is not limited thereto.

[0025]

Example 1: Extracellular enzyme activity of KI2N strain KI2N strain and Bacillus licheniformis IFO12199, Bacillus lic
The extracellular enzyme activity of heniformisMN-001 (FERM BP-266, which is regarded as a strain having an excellent extracellular enzyme-producing ability) was compared. KI2N strain and Bacillus licheniformis I
The method of measuring the enzyme activity of FO12199 is "Biochemistry learned through experiments"
According to That is, using the culture supernatant of the microorganism as a crude enzyme solution,
Amylase activity was measured by the DNS method using the substrate as starch, cellulase activity was measured by the DNS method using the substrate as CMC, and protease activity was measured by using the Anson-Hagiwara method and the Lowry method with the milk casein as the substrate. Table 1 shows the results. As a result, it was revealed that the KI2N strain exhibited high extracellular enzyme activity.

(Table 1: Extracellular enzyme activity) ア ミ Amylase Cellulase Protease ──────────────────────────────────── KI2N strain 14.0 13.9 60.7 B. licheniformis MN-001 2.0 15.2 9.8 B. licheniformis IFO12199 1 1 1 ──────────────────────────────────── The relative value when the enzyme activity of licheniformis IFO12199 is set to 1. The value of Bacillus licheniformis MN-001 is based on Japanese Patent Publication No. 3-79986.

[0027]

[Example 2: Fungal growth inhibitory action]
The fungal growth inhibitory effect of the 2N strain was tested. That is, one loopful of the KI2N strain was inoculated into 50 ml of a normal broth medium and cultured at 37 ° C. for 24 hours. 25 μl of the culture solution
It was adsorbed on a paper disk and placed on two places (B) of a PDA agar medium. In addition, the agar medium on which the test fungus was cultured was punched out together with the fungus with a 6φ cork borer, and placed at one location (A) between the above B (however, Aspergillus ni
ger spores were scattered with McFarland No. 1 using Tween 80 plus 1% saline to prevent spores from scattering. The suspension was adjusted to a suspension degree of 2 and dropped at one place (A)). As a control, KI2
Instead of the N strain, a medium in which two paper disks (B) adsorbing 25 μl of a physiological saline solution were used. Table 2 shows the results. As a result, it was found that the KI2N strain had an action of suppressing the growth of many fungi.

(Table 2: inhibitory effect on fungal growth) に よ る Depends on test fungi strain KI2N Growth inhibition ───────────────────────────────── Aspergillus flavus + Aspergillus fumigatus + Aspergillus niger JCM 2261 + Aspergillus ochraceus + Aureobasidium pullulans IFO 6353 ＋ Botrytis cinerea IFO 31831 ＋ Cheatomiun globosum IFO 6347 ＋ Cladosporium cladosporioides IFO 6348 ＋ Fusarium oxysporum f. sp. Rosellinia necatrix + Rosellinia necatrix IFO 9420 + Valsa ceratosperma + Valsa ceratosperma IFO 30252 + ─────────────────────────────────

Many of the above fungi have pathogenicity to plants (those without accession numbers are field isolates).

[0030]

Example 3 Fermentation-Promoting Action Fresh cow manure of lactating Holstein cows (leaf litter) was transferred to a burn cleaner or a compost house and deposited, and such deposits were used as test materials.
At this point, the moisture content of the sediment was about 80%, and the shred length of the hay was 45 cm. The composting room consists of a test plot and a control plot.
A plot was set up, and 1 × 10
⁹ CFU / g KI2N strain (cultured by a liquid culture method, heated to form spores, then washed three times by centrifugation using a physiological saline solution, the components of the culture solution were removed, and adjusted with vermiculite. Was added and mixed. The control group was not added. The sediment was turned over on both test plots on the same day.

The progress of fermentation was observed using the temperature change of the sediment and the length of the hay included in the sediment (average value of 50 randomly selected lengths) as indicators. 4
Was obtained.

As is evident from these results, in the test plot, the temperature was raised quickly and continuously, and the decomposition of pasture hay remarkably progressed. In the test plot, the color tone of the sediment changed remarkably. After 5 days from the start of the test, although the central part of the fecal mass was partially green, the black color increased, and after 17 days, only a small part of the lower layer was observed. , Except that the color was completely black after 30 days, but almost no change in color tone was observed in the control group. From these results, excellent fermentation promotion of the KI2N strain was confirmed. Conventionally, composting organic matter has taken a long time. For example, pasture hay has been ripened for at least three months, and more than one year for long.

(Table 3: Temperature change of sediment (° C.)) 後 After start of test Number of days (A: cut back) {0 1 3 5A 6 7A} ───────────────────────── Control section 21 21 23 23 25 21 ───────────────── ────────────── Test zone 21 47 85 82 75 82 ──────────────────────────── ───

(Table 3: Temperature change of sediment (° C.): continued) 数 Days after start of test ( A: cut back) {13 15 21A 25 30}対 照 Control plot 25 25 23 23 22 ─────────────────────────── Test plot 78 82 76 55 46 ───────────────────────────

(Table 4: Length of pasture hay (cm)) 日 Days after start of test ─── ──────────────────────── 0 10 30 ───────────────────────対 照 Control plot 45 45 45 ─────────────────────────── Test plot 45 254 ───────── ──────────────────

[0036]

Example 4: Effect of organic fertilizer added with KI2N strain on leeks Leek (variety: Konatsu, Takii seedling) was used as a test crop and horticultural soil (composed of red clay 50: peat moss 25: vermiculite 10: charcoal 15%, component N200) : P ₂ O ₅ 15
00: K ₂ O 200: MgO 100 mg / l) is used as a test soil, and rapeseed meal equivalent to about 1% of the test culture soil has an amount of K that is about 10 ⁷ CFU / g per 1 g of the test soil dry weight.
An organic fertilizer to which an I2N strain (cultured by a liquid culture method, heated to form spores, and then washed three times by centrifugation using a physiological saline solution to remove the culture solution components) was provided. It was a fertilizer. A control plot was obtained by adding only rapeseed meal equivalent to about 1% of the test soil and the test culture soil to the No. 5 plastic pot, and a test plot was obtained by adding the test soil and the test fertilizer to the test plot. A test for sowing 20 test seeds and observing growth was carried out twice. Table 5 shows the obtained results.

As is clear from the results, 15
On the other hand, no significant difference was observed in the height of the leek between the two test sections, but the onion in the test section continued to grow continuously, and the height, root length and foliage weight were measured on the 55th day. -Remarkable growth of the administration group was confirmed using the root weight as an index. These results confirmed a sustained effect when used as a KI2N strain fertilizer additive.

(Table 5: Growth Status of Leek) 計 測 Measurement Items Days after Seeding (Sun) 15 30 55 ───────────────────────────────── Control plot Plant height (cm) 3.2 8. 4 15.9 Root length (cm) 16.1 Foliage weight (g) 6.88 Root weight (g) 3.78 ────────────────────── ─────────── Test plot Plant height (cm) 3.0 8.2 24.6 Root length (cm) 18.5 Foliage weight (g) 20.65 Root weight (g) 10.75 ─────────────────────────────────

[0039]

[Example 5: Effect of organic fertilizer added with KI2N strain on spinach] Spinach (cultivar: Atlanta) was used as a test crop, and light-colored andoso soil was used as a test soil.
The amount of fermented chicken manure equivalent to 00 kg is about 10 ⁷ CFU / g per 1 g of soil per 15 cm of test soil soil (cultured by the liquid culture method and then heated to form spores, then physiological saline The organic fertilizer to which the liquid was washed three times by centrifugation to remove the components of the culture solution) was used as a test fertilizer. Tunnel covering the field,
A plot in which only fermented chicken manure was added to the test soil was set as a control plot, and a plot in which the test soil and the test fertilizer were added was set as a test plot, and seeds of the test crop were sown at a substantially uniform density. Leaf length and number of developed leaves were measured at three locations, 10 for each plot, yield (biological weight)
Was measured at three locations of 1 m ^{2 in} each section, and the average value was determined. Table 6 shows the obtained results.

As is clear from the results, 15
No significant difference was observed in the leaf length between the two test sections until day 30. The spinach in the test section continued to grow continuously. As an index, remarkable growth was confirmed in the test plot compared to the control plot. These results confirmed a sustained effect when the KI2N strain was used as a fertilizer additive.

(Table 6: Growth status of spinach) ───────────────────────────────── Measurement items Days after sowing (Sun) 15 30 60 対 照 Control section Leaf length (cm) 3.2 12 2.6 23.2 Number of developed leaves (sheets) 2.5 Yield (g) 24.3 ───────────────────────────── ──── Test section Leaf length (cm) 3.6 11.8 29.3 Number of developed leaves (sheets) 3.6 Yield (g) 39.2 ────────────── ───────────────────

[0042]

Example 6: Inhibitory Effect of Seedling Blight on Cucumber Cucumber (variety: Akishimari Midori, Tohoku Co., Ltd.) was used as a test crop, and commercially available black soil sterilized in an autoclave was used as a test soil. In addition, the mixture of soil 9: rice bran 1
After adjusting to 0%, put in a kolben, sterilize by autoclave at 121 ° C for 15 minutes, cool to 25 ° C, and then
Four groups of octonia solani ag were inoculated, cultured at about 27 ° C. for one week, then ground well in a mortar, mixed well with 49 g of the test soil, and stirred to obtain a pathogen culture medium. For the test soil,
0.1% pathogen culture medium and 10 ⁸ C / g of test soil
FU / g KI2N strain (cultured by liquid culture method, heated to form spores, and then washed three times by centrifugation using a physiological saline solution to remove the culture solution components) The section to which was added was used as a test section. A section in which only 0.1% pathogen culture medium was mixed with the test soil was used as a control section. In each of the test plots, 200 g each of the soil prepared as described above was put into a pot, and five test crops were sown. Tables 7 and 8 show the obtained results (suppression of seedling blight of vegetables and changes in the number of pathogenic fungi and KI2N in soil), respectively.

As is clear from the results, the seeding 7
Observation after day, the control group showed obvious germination failure and abnormal growth, but in the test group, in each of the pots, only a small wound was observed at the boundary between the aboveground part and the underground part, It seemed to prevent the onset of the disease. Since the number of pathogenic bacteria in the soil was reduced in the test plot, it was shown that the KI2N strain not only suppressed the growth of pathogenic fungi but also killed the pathogenic fungi.

(Table 7: Suppression of vegetable seedling blight by KI2N strain (total score for each pot)) Number of repeats 1 2 3 average 対 照 Control section 12 18 15 15.0 区───────────────────── Test zone 103 ───────────────────── ────── Evaluation score 0: No abnormalities 1: Small holes or scratches on stems 2: Narrowing at the border between above ground and underground 3: Narrowing between above ground and underground Seedlings fall 4: Root extends first, head is on the ground (handstand) 5: Does not germinate

Table 8: Changes in the number of pathogenic fungi and the number of KI2N strains in soil (CFU / g)時 At the start of the test At the end of the test ─────────────────────────────── Control section Rhizoctonia solani 3 × 10 ⁵ 4 × 10 ⁵ KI2N strain 10 ² ＞ 10 ² ＞ ─────────────────────────────── Test area Rhizoctonia solani 5 × 10 ⁵ 10 ² > KI2N strain 8 × 10 ⁷ 7 × 10 ⁷は, Average of 3 pots in each ward

[0046]

Example 7: Soil improvement effect (1) The soil improvement effect of the KI2N strain on soil contaminated with phytopathogenic fungi was confirmed as follows.

The soil in the test section and the control section was adjusted by the method according to the example shown in Example 6. That is, the soil in the test plot is black soil containing Rhizoctonia solani and KI2N strain, and the soil in the control plot is black soil containing Rhizoctonia solani but not KI2N strain. In each plot, 200 g of soil was put into a pot and Rhizoctonia solani and K
The number of bacteria of the I2N strain was measured. Table 9 shows the obtained results. As a result, in the control plot, 3 to 7 ×
Only Rhizoctonia solani was detected in the range of 10 ⁶ CFU / g. In the test plot, 7 days after the start of the test, Rhizoctonia
The number of solani cells was below the detection limit, and was not detected thereafter. On the other hand, the KI2N strain was always detected in the range of 5 to 9 × 10 ⁸ CFU / g throughout the test period. The results showed that the KI2N strain not only suppressed the growth of the pathogenic fungus, but also killed the pathogenic fungus.

(Table 9: Changes in the number of pathogenic fungi and the number of KI2N strains in soil (CFU / g))日 Days after start of test 0 7 14 21 ──────────────────────────────── ──── Control area Rhizoctonia solani 7 × 10 ⁶ 5 × 10 ⁶ 3 × 10 ⁶ 7 × 10 ⁶ KI2N strain 10 ² > 10 ² > 10 ² > 10 ² > ──────────── ──────────────────────── Test plot Rhizoctonia solani 6 × 10 ⁶ 10 ² > 10 ² > 10 ² > KI2N strain 7 × 10 ⁸ 9 × 10 ⁸ 5 × 10 ⁸ 6 × 10 ⁸ ────────────────────────────────────

[0049]

Example 8: Soil improvement effect (2) The soil improvement effect of the KI2N strain on soil contaminated with phytopathogenic fungi was confirmed as follows.

The pathogen culture medium prepared in the same manner as in Example 6 was added to each pot of the control plot and the test plot after 21 days in Example 7 in an amount of 0.1% with respect to the soil of the pot. After such additional inoculation, it is again over time for 21 days
The numbers of Rhizoctonia solani and KI2N strains were measured. Table 10 shows the obtained results. As a result, in the control plot, Rhi ranged from 8 to 10 × 10 ⁶ CFU / g throughout the test period.
Only zoctonia solani was detected. In the test plot, the number of Rhizoctonia solani cells was below the detection limit 7 days after the additional inoculation, and was not detected thereafter. on the other hand,
The KI2N strain was 3 to 7 × 10 ⁸ CFU /
g was always detected. The results showed that the inhibitory effect of the KI2N strain on pathogenic fungi was persistent.

(Table 9: Changes in the number of pathogenic fungi and the number of KI2N strains in soil (CFU / g))日 Days after booster vaccination 0 7 14 21 ──────────────────────────────── ──── Control area Rhizoctonia solani 1 × 10 ⁷ 8 × 10 ⁶ 8 × 10 ⁶ 9 × 10 ⁶ KI2N strain 10 ² ＞ 10 ² ＞ 10 ² ＞ 10 ² ＞ ──────────── ──────────────────────── Test area Rhizoctonia solani 5 × 10 ⁶ 10 ² > 10 ² > 10 ² > KI2N strain 5 × 10 ⁸ 7 × 10 ⁸ 3 × 10 ⁸ 4 × 10 ⁸ ────────────────────────────────────

[0052]

Example 9 One platinum loop of KI2N strain (F
ERM P-17147) and inoculated at 37 ° C. with stirring for 48 hours. After culturing, the mixture was heated to form spores, washed with physiological saline, and the culture solution was removed. The KI2N strain culture thus obtained (10 ⁵ to 10 ¹⁰ C
FU / g) 1 part by weight, diatomaceous earth 1 part by weight and clay 1
After uniformly mixing and drying parts by weight, pulverized, and powdered KI2
An N-strain-containing composition was produced. This composition was particularly suitable as a soil improving composition and an antifungal composition.

[0053]

The Bacillus cereus KI2N of the present invention
The strain and the fermentation promoter composition, the fertilizer additive composition, the soil conditioner composition, and the fungal growth inhibiting composition using the same contribute to the recycling of organic waste, improve the soil, and improve the plant. Control plant disease and promote plant growth. Sustained effects promote the implementation of recycling-oriented agriculture with reduced use of chemical substances such as fertilizers and pesticides. There is no problem with safety.

The details of the mechanism of action will need to be repeated in the future, but at present it is estimated as follows.

The KI2N strain has a high extracellular enzyme activity. These enzymes seem to promote the fermentation of organic matter. In addition, conventional fermentation promoters utilizing microorganisms were effective in the early stage of fermentation but did not last long, but KI2N
The reason why the effect of the strain is persistent is that the KI2N strain is a facultative anaerobic bacterium, and even though the initial fermentation proceeds and some organic substances become anaerobic, the metabolism is slightly inferior when aerobic. What can be done is working advantageously.

Also, the KI2N strain has many kinds of fungi,
Among them, it has a growth inhibitory effect on phytopathogenic fungi. It is considered that the improvement of soil contaminated with phytopathogenic fungi and the suppression of pathogenic fungi on the plant surface involve the growth inhibitory action against the phytopathogenic fungi.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI C05F 11/08 C05F 11/08 C09K 17/32 C09K 17/32 H C12N 1/00 C12N 1/00 S // (C12N 1 / 20 C12R 1: 085)

Claims (10)

(57) [Claims] 1. A Bacillus cereus KI2N strain (FERM P-) having the following mycological properties:
17147). Cell swelling-NaCl 7% + Anaerobic growth + Starch degradation + VP-Egg-yolk-Indole-Growth at 60 ° C-Gram staining + Spore position Center and semi-end position Lusterless Colony surface Wrinkles Cell size 1.2x 3-3.5 μm DHL-MacConkey-mannitol medium-normal agar medium + 10% sheep blood-supplemented blood agar medium + catalase + urea degradation-oxidase + hemolytic + hydrogen sulfide-nitrate reduction + ONPG-acid production from carbohydrate Galactose-D-mannose-melibiose-xylose-L-arabinose-glucose + mannitol-rhamnose-glycerol-esculin + extracellular enzyme activity High organic matter fermentation promoting action + plant growth promoting action + fungal growth inhibitory action + continuous action + 2. The Bacillus cereus KI2N strain according to claim 1 (FERM P-17).
147) or a processed product thereof. 3. The composition is at least one selected from a fermentation promoter composition, a fertilizer or a fertilizer additive composition, a soil conditioner composition for disinfecting contaminated soil, and an antifungal composition. The composition according to claim 2, characterized in that: 4. The fermentation promoter composition according to claim 3,
A fermentation method for organic matter, characterized by sensitizing organic matter. 5. An organic agricultural material produced by the fermentation method according to claim 4 using an organic substance. 6. A fertilizer comprising the fertilizer composition according to claim 3 or a fertilizer additive composition added thereto. 7. A method for cultivating a plant, comprising cultivating by applying the fertilizer according to claim 6. 8. A method for improving disinfection of contaminated soil, which comprises applying the soil conditioner composition according to claim 3 to soil. 9. A method for disinfecting or inhibiting the growth of phytopathogenic fungi, which comprises applying the antifungal agent composition according to claim 3 to a plant. 10. The method of applying the antifungal agent composition to a plant, which comprises coating the seed of the plant, mixing with the cultivation soil of the plant, or spraying on the foliage of the plant.
The method for sterilizing or inhibiting the growth of phytopathogenic fungi according to the above.