JPH08175920A - Agricultural and horticultural germicidal composition - Google Patents

Abstract

PURPOSE: To obtain an agricultural and horticultural germicidal composition, excellent in disease injury controlling actions and good in colonization properties and persistence stability by utilizing a bacterium of the genus Bacillus. CONSTITUTION: This agricultural and horticultural germicidal composition contains spores of a bacterium belonging to the genus Bacillus, preferably a bacterium Bacillus subtilis FERM P-14647 strain or Bacillus pubtilis FERM P-14646 strain and a humectant, preferably polysaccharides, oligosaccharides or synthetic water-soluble polymers. The spores are obtained by carrying out the solid culture of the bacterium belonging to the genus Bacillus or the liquid culture thereof under aerobic conditions, centrifuging the resultant cultured product and removing the culture supernatant. The content of the spores in the germicidal composition is preferably 0.001-99.9 pts.wt. and that of the humectant is preferably 0.1-99.999 pts.wt. based on the total amount of the composition. Methods for coating seeds therewith, treating a single flower, treating leaves or stems, coating a wound site or a pruned part of the cultivated plants, irrigating soil or mixing soil therewith, etc., are cited as the method for applying the composition.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an agricultural / horticultural fungicide composition, and more particularly to an agricultural / horticultural fungicide composition containing a spore of a bacterium belonging to the genus Bacillus and a moisturizer and having an excellent disease control action. .

[0002]

2. Description of the Related Art In recent years, as a method of protecting agricultural and horticultural plants from various diseases, in consideration of safety and sustainability of effects, the occurrence of diseases is prevented by using microorganisms that compete with pathogenic bacteria that cause various diseases. The method is widely used.

Microorganisms that have been used for controlling the diseases of such agricultural and horticultural plants include bacteria belonging to the genera Trichoderma, Gliocladium, Bacillus, and the like, and farms containing these microorganisms so far. Many horticultural germicide compositions have also been researched and developed.

Regarding the bacteria belonging to the genus Bacillus among them, for example, in JP-A-63-273470, Bacillus subtilis JB3 strain (NCIB12375), Bacillus subtilis JB
Antibacterial substances obtained from the 3.6 strain (NCIB12376), Bacillus subtilis R1 strain (NCIB12616) or mutants thereof suppress plant diseases, microbial infections of animals and humans, and further prevent general microbial contamination. As a control, attempts have been made to control diseases of various agricultural and horticultural plants using cultures of the above strains.

Further, in Japanese Patent Laid-Open No. 22222/1990,
Bacillus subtilis JB above
Antibiotics derived from 3 strains (NCIB12375), Bacillus subtilis JB3.6 strain (NCIB12376), Bacillus subtilis R1 strain (NCIB12616) or mutants thereof are isolated, and these are used as a disease of various agricultural and horticultural plants. Attempts have been made to use it for control.

[0006] However, none of the methods of controlling diseases of agricultural and horticultural plants using such bacteria belonging to the genus Bacillus is not always sufficient in terms of sustainability, fixability and stability.

[0007] It is known that such bacteria belonging to the genus Bacillus form spores, and spores are often isolated from cultures. No examples have been reported so far, which have been prepared in a large amount, or cases where spores were combined with a moisturizer to try to show the effect of controlling diseases on agricultural and horticultural plants.

[0008]

The present invention has been made from the above point of view, and in an agricultural and horticultural fungicide composition utilizing a bacterium belonging to the genus Bacillus, excellent in disease control action, and further, fixability, It is an object of the present invention to provide a fungicide composition for agricultural and horticultural use with good long-term stability.

[0009]

Means for Solving the Problems As a result of intensive research to solve the above problems, the present inventor took out spores from a culture of a bacterium belonging to the genus Bacillus and sterilized them with a moisturizer for agricultural and horticultural purposes. It was found that an agricultural and horticultural fungicide composition having an excellent disease control action and having good fixability and long-term stability can be obtained by adding it to the agent composition, and completed the present invention.

That is, the present invention is an agricultural and horticultural fungicide composition containing spores of a bacterium belonging to the genus Bacillus and a moisturizer. Hereinafter, the present invention will be described in detail.

<1> Bacterial spores belonging to the genus Bacillus The agricultural / horticultural fungicide composition of the present invention contains spores obtained from a culture of bacteria belonging to the genus Bacillus as an active ingredient.

The bacterium belonging to the genus Bacillus used in the present invention is not particularly limited as long as it is a bacterium belonging to the genus Bacillus, but preferably a bacterium belonging to the genus Bacillus which competes with a plant pathogenic bacterium, among which more preferred Include Bacillus subtilis, and among them, Bacillus subtilis FERM
P-14647 strain, Bacillus subtilis FERM
P-14646 strain is preferable.

Bacillus subtilis FERM P-14
647 strain and Bacillus subtilis FERM P-1
The strain 4646 has been deposited with the Institute of Biotechnology, Industrial Technology Institute (FERM) on November 17, 1994. Also, Bacillus subtilis FERM P-14
Strain 647 is NCIB12376 strain, Bacillus subtilis FERM P-14646 strain is NCIB1 strain
As strain 2616, Scotland, AB98DG,
Aberdeen, Abbey Ro
ad) 135, P.I. O. Box 31 National Collections of Industrial and Marine Bacteria
and Marine Bacteria Ltd.) (NCIB), 1 at Torry Research Station
December 22, 986 (NCIB12376), 19
Deposited on Dec. 24, 1987 (NCIB12616), the nature of these strains is described in European Patent Application Publication No. 2
No. 76132.

The spores used in the present invention are obtained from a culture of the bacterium belonging to the genus Bacillus. Cultivation of bacteria belonging to the genus Bacillus can be carried out in accordance with ordinary culturing methods of bacteria belonging to the genus Bacillus, for example, liquid culture such as reciprocal shaking culture, jar-famenter culture, culture tank culture, and solid culture. .

The medium used for the culture may be any medium as long as it can easily form spores efficiently. As carbon sources, sugars such as glucose, starch, dextrin, sucrose and molasses, organic acids such as citric acid and malic acid, Alcohols such as glycerin are used as nitrogen sources such as ammonia, ammonium sulfate, ammonium chloride, ammonium salts such as ammonium nitrate and nitrates and yeast nitrogen, corn steep leaker, meat extract, wheat germ, polypeptone, soybean flour and other organic nitrogen sources. As the inorganic salt, salts of phosphoric acid, potassium, calcium, manganese, magnesium, iron and the like, for example, potassium chloride, calcium chloride, manganese sulfate, ferrous sulfate and the like can be added. Further, various additives such as an antifoaming agent can be used if necessary.

The culture conditions are not particularly limited, but the culture is preferably solid culture or liquid culture performed under aerobic conditions such as aeration and shaking culture, and the temperature is preferably 10 to 50. C, more preferably 15-40
C. and pH are preferably in the range of 4 to 9, more preferably 6 to 8.

The method for separating spores from the culture belonging to the genus Bacillus obtained as described above can be carried out by a method such as membrane separation, centrifugation or filtration separation.
The resulting spore fraction may be used as it is in the agricultural and horticultural fungicide composition described below in a state of containing a certain amount of water, and if necessary, a drying method such as freeze drying, ventilation drying, and spray drying. It can also be used as a dried product in an agricultural / horticultural germicide composition.

A method for producing spores of a bacterium belonging to the genus Bacillus used in the present invention is described in, for example, Bacillus subtilis F.
The case of using the ERM P-14647 strain will be described. Bacillus subtilis FERM P-14647
The slope culture of the strain is inoculated into a Sakaguchi flask containing broth medium (meat extract 1%, peptone 1%, NaCl 0.5%), and then cultured at 30 ° C. for 1 day on a rotary shaker. The obtained culture was used as a medium (glucose 2%, polypeptone 1
%, Yeast extract 0.2%, KH ₂ PO ₄ 0.1%) was inoculated into the fermenter and cultured under aerobic conditions at 30 ° C. for 72 hours to obtain a culture solution. This culture solution is centrifuged to separate it into a culture supernatant and a bacterial cell precipitate, and after removing the culture supernatant, the bacterial cell precipitate is washed with water to obtain wet bacterial cells (spore fraction). Further, this spore fraction (wet cell) is frozen at −80 ° C., dried under reduced pressure and pulverized to give a spore fraction as a dried product. Bacillus subtilis FE obtained in this way
The spore fraction of the RM P-14647 strain culture contains the spores of the strain in an amount of about 50% to 100% by dry weight.

<2> Moisturizer A moisturizer is added to the agricultural / horticultural germicide composition of the present invention together with the spores of the bacteria belonging to the genus Bacillus.

The moisturizer used in the present invention is not particularly limited as long as it is generally used as a moisturizer in food additives and the like, and examples thereof include gum arabic, pullulan,
Gellan gum, xanthan gum, soabi gum, natural polysaccharides such as alginic acid and oligosaccharides, inorganic substances such as aluminum silicate, polyhydric alcohols such as polyethylene glycol, propylene glycol and glycerol, and their polymers, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acid, etc. Synthetic water-soluble polymers, cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, hydroxyethylmethylcellulose, sucrose fatty acid ester in which sucrose and fatty acids of various oils and fats are ester-bonded, pyrophosphate, sodium tripolyphosphate, sodium pyrophosphate , Phosphoric acid such as sodium metaphosphate, and phosphates.

Among the above compounds, polysaccharides, oligosaccharides, synthetic water-soluble polymers and the like are preferably used in the present invention. Further, in the agricultural and horticultural germicide composition of the present invention, one of these compounds may be blended alone as a moisturizing agent component, or two or more of these compounds may be blended and blended. Is.

All of the above compounds are commercially available, and are generally easily available. An example of a commercially available product is oligosaccharides as Fuji Oligo from Nippon Shokuhin Kako Co., Ltd.
, A water-soluble cellulose ether, which is a cellulose derivative, is marketed by Shin-Etsu Chemical Co., Ltd. as METOLOSE, and a sucrose fatty acid ester is marketed by Dai-ichi Kogyo Seiyaku Co., Ltd. as DK ester.

<3> Agro-horticultural fungicide composition The agricultural-horticultural fungicide composition of the present invention contains one or more kinds of spores and moisturizers of bacteria belonging to the genus Bacillus.

When the spores of the bacteria belonging to the genus Bacillus are blended with the fungicide composition for agricultural and horticultural use of the present invention, the weight of the spores contained should be 0.001 to 99.9 parts of the total weight of the composition. It is preferable to mix it with Spore content is 0.00
If it is less than 1 part by weight, the disease controlling effect may not be sufficient, and if it exceeds 99.9 parts by weight, an effective amount of the moisturizing agent cannot be formed in the agricultural and horticultural bactericidal composition, and the disease controlling effect is also expected. May not be enough.

The amount of the moisturizing agent is preferably 0.1 to 99.999 parts by weight with respect to the total amount of the composition. If the content of the moisturizing agent is less than 0.1 part by weight, excellent disease control action may not be obtained, and if it exceeds 99.999 parts by weight, a sufficient amount of spores to exert the disease control action may be obtained. Unable to be formulated in an agricultural and horticultural germicidal composition,
An excellent disease control effect due to spores may not be obtained.

The fungicidal composition for agricultural and horticultural use of the present invention comprises a spore of a bacterium belonging to the genus Bacillus and a moisturizer together with various optional components, if necessary, in the form of powder, granules, granules, Wettable powder, emulsion, liquid, flowable,
It is formulated into a coating agent or the like.

The above-mentioned optional components include solid carriers,
Carion clay, pyrophyllite clay, bentonite, montmorillonite, diatomaceous earth, synthetic hydrous silicon oxide,
Fine mineral powder such as acid clay, talc, clay, ceramics, quartz, sericite, vermiculite, perlite, Otani stone, anthra stone, limestone, coal ash, zeolite, etc.
Inorganic salts such as potassium salt, calcium salt (including shell powder), manganese salt, magnesium salt, iron salt, ammonium salt (ammonium chloride, ammonium sulfate, ammonium nitrate, etc.), rice straw, rice husk, bran, chitin (crab shell, (Including shrimp shells, chitin-containing fine powders such as krill), rice bran, wheat flour, corn cobs, peanut shells, bone meal, fish meal, meal meal, sawdust, wood meal, charcoal, charcoal, bark charcoal,
Rice husk charcoal, plant charcoal, peat moss, attapulgite,
Examples thereof include dry feces, activated carbon, oil cake, and fine powder of organic matter such as soybean powder. Further, as the liquid carrier, water,
Vegetable oil, liquid animal oil, etc. are mentioned. Furthermore, as an auxiliary agent, if necessary, a soluble extender (starch hydrolyzate,
It is also possible to use D-sorbitol, lactose, maltose, etc.), fixing agents and dispersants, antifreezing agents, thickeners and the like.

The plant pathogens to which the fungicide composition for agricultural and horticultural use of the present invention thus obtained is applied, those pathogens belonging to molds, for example, Rhizoctonia solani, a rice wilt disease fungus. ), The blast fungus Pyricularia oryzae, and the powdery mildew fungus Erysiphe graminis of barley.
graminis), wheat wilt fungus Geumanomyces graminis, pea brown spot fungus Ascochyta pisi, broad bean red spot fungus Botrytis fabae (Botrytis faba)
e),

Vegetables such as tomato, eggplant, strawberry,
Botrytis cinerea, a fungus that causes gray mold such as cucumber, lettuce, and green beans, and Alternaria brass
icicola), tomato leaf mold fungus Cladosporium fulvum, phytophthora infestans Phytophthora infestans, Fusarium oxysporu
m), powdery mildew fungus of cucumber Sphaerotheca fuliginea, downy mildew fungus Pseudoperonospora cubensi
s), wilt fungus Pythium sp., rust rust fungus Puccinia allii, Sclerotinia allii
Chinese cabbage leaf spot fungus Alternaria brassicae (Altern
aria brassicae), carrot black leaf blight fungus Alternaria dauci, spinach wilt fungus Rhizoctonia solani, lettuce sclerotinia sclerothiolam (Sclero)
tinia sclerotiorum), Verticillium dahliae, a half-body wilt fungus of eggplant, and powdery mildew fungus Sphaerotheca of strawberry.
humuli), anthracnose fungus
etotrichum fragariae),

[0030] Flowers such as cyclamen, chrysanthemum, rose, and statice Botrytis cinerea, powdery mildew fungus Sphaerotheca pannosa, white rust fungus Puccinia holiana (Puccinia) horiana),

[0031] Pseudomonas aeruginosa (Rosellinia necatrix) on fruit trees, Penicillium italicu (Penicillium italicu) on citrus
m), Black spot bacterium Diaporthe citr
i), Gymnosporangium asiaticum, a red scab of pear, Venturia inaequali, a black scab of apple, Venturia inaequali
s), Peach scab, Monilinia fructicola (Mon
ilinia fructicola),

[Large patch disease fungus Rhizoctonia solani (Rhizoctonia solani) of the lawn, leaf blight fungus curvularia
sp (Curvularia sp), Helmint sporium sp
(Helminthosporium sp), rust fungus Puccinia zoysiae, dollar spot fungus Sclerotinia homeocarpa (Sclerotinia homoeocarp)
a), spring blight fungus Fusarium, Rhizoctonia, Pythium, snow rot Typhula incarnata, brown patch fungus Rhizoctonia sola
ni) and the like.

<4> Method for Applying Agricultural and Horticultural Fungicide Composition of the Present Invention The agricultural and horticultural fungicide composition of the present invention is applied to cultivated plants for the purpose of controlling various diseases of the various cultivated plants as described above. However, the method is appropriately selected depending on the usage form such as dosage form, crop and disease, and for example, above-ground liquid spray, above-ground solid spray, aerial liquid spray, aerial solid spray, water surface application, facility application, soil mixing. Examples thereof include application, soil irrigation application, surface treatment (seed dressing, application treatment, etc.), nursery box application method, single flower treatment, and plant base treatment. Further, among these application methods, preferably, the seeds of cultivated plants are coated with various dosage forms of agricultural and horticultural fungicide compositions, the flowers of cultivated plants are subjected to single flower treatment, the foliage of cultivated plants is treated, Examples of the method include applying treatment to the wound site and pruning part of the cultivated plant, irrigating the soil, and mixing the soil.

In addition, when applying the agricultural and horticultural fungicide composition to cultivated plants, insecticides, nematicides, acaricides, herbicides, fungicides, plant growth regulators, fertilizers, soil improving materials (peat) , Humic acid materials, polyvinyl alcohol materials, etc.) may be mixed and applied, or they may be alternately applied without mixing or simultaneously applied.

The application rate of the fungicide composition for agricultural and horticultural use of the present invention varies depending on the type of disease, the type of plant to be applied, the dosage form of the fungicide composition, etc., but cannot be specified unconditionally. When the horticultural germicide composition is sprayed on the ground, the spore concentration for application is usually about 10 ⁶ CFU (colony forming unit) / mL to ^{10 10} CFU / mL, preferably about 10 ⁷ CFU / mL. -10 ⁹ CFU / mL, application rate is 0.5-50 L / a. In addition, granules, powders, etc. can be applied as they are without any dilution, and when sprayed on the ground, the spore application rate is about 10 ¹² to 10 ¹⁴ CFU / a. Preferably.

[0036]

Embodiments of the present invention will be described below. First, an example of the production of spores of a bacterium belonging to the genus Bacillus that is added to the agricultural / horticultural germicide composition of the present invention will be described.

[0037]

[Production Examples 1 and 2] Spore fraction of a bacterium belonging to the genus Bacillus (wet cell) One platinum loop of a slope culture of a preservation bacterium of Bacillus subtilis FERM P-14647 strain is 100 m per flask.
L broth medium (meat extract 1%, peptone 1%, Na
Sakaguchi flask containing 500% Cl (500 mL)
After inoculation, the cells were cultured for 1 day at 30 ° C. using a reciprocal shaker with an amplitude of 10 cm and a rotation speed of 120 rpm. 300 mL of the obtained culture was used as a medium (glucose 2%, polypeptone 1%, yeast extract 0.2%, KH ₂ PO ₄ 0.1%).
A 30 L fermenter containing L was inoculated and cultured under aerobic conditions at 30 ° C. for 72 hours to obtain a culture solution. Obtained about 1
Centrifuge 5 liters of culture broth according to the standard method (6000 rp
m, 20 minutes) to separate the culture supernatant and the bacterial cell precipitate.
After separating the culture supernatant as an antibiotic fraction (Comparative Example 1), the bacterial cell precipitate was washed with water to obtain a wet weight of about 780 g of wet bacterial cells (spore fraction) (Production Example 1). This spore fraction contains 100% by dry weight of spores of Bacillus subtilis FERM P-14647.

In the above, Bacillus subtilis FERM P
Instead of using the 14647 strain, the same operation was performed using the Bacillus subtilis FERM P-14646 strain, and a wet weight of about 750 containing 50% by dry weight of spores of the Bacillus subtilis FERM P-14646 strain was used.
A spore fraction of g was obtained (Production Example 2).

[0039]

[Comparative Example 2] Viable cell fraction of bacteria belonging to the genus Bacillus (wet cells) One platinum loop of a slope culture of a preservation bacterium of Bacillus subtilis FERM P-14647 strain was 100 m per flask.
L broth medium (meat extract 1%, peptone 1%, Na
Sakaguchi flask containing 500% Cl (500 mL)
After inoculation, the cells were cultured for 1 day at 30 ° C. using a reciprocal shaker with an amplitude of 10 cm and a rotation speed of 120 rpm. 300 mL of the obtained culture was used as a medium (glucose 2%, polypeptone 1%, yeast extract 0.2%, KH ₂ PO ₄ 0.1%).
A 30 L fermenter containing L was inoculated and cultured under aerobic conditions for 18 hours to obtain a culture solution. About 15 L of the obtained culture solution was centrifuged (6000 rpm, 20 minutes) according to a conventional method to separate into a culture supernatant and a bacterial cell precipitate. After removing the culture supernatant, the microbial cell precipitate was washed with water to obtain a wet microbial cell (viable microbial cell fraction) having a wet weight of about 600 g. This viable cell fraction contains 3% by dry weight of spores of Bacillus subtilis FERM P-14647 strain.

[0040]

[Production Examples 3 and 4] Powder spores Bacillus subtilis obtained in the above Production Example 1
About 600 g of the spore fraction of the FERM P-14647 strain was suspended in 3 L of water and then treated with a spray dryer (manufactured by Niro Japan) at a flow rate of 1.5 to 2 L / hr (inlet temperature 150 ° C., outlet temperature 100 ° C.). . The dried product obtained by the spray dryer was pulverized to obtain a powder spore having a dry weight of about 78 g (Production Example 3).

Similarly, about 78 g of Bacillus subtilis FERM was obtained from about 600 g of the spore fraction obtained in Production Example 2.
A powder spore of P-14646 strain was obtained (Production Example 4).

[0042]

Comparative Example 3 Viable Cell Suspension Bacillus subtilis Obtained in Comparative Example 2
About 100 g of the viable cell fraction of the FERM P-14647 strain was suspended in 10 L of water to obtain a viable cell suspension.

Next, examples of the fungicide composition for agricultural and horticultural use of the present invention containing the spore fraction of the bacterium belonging to the genus Bacillus obtained in each of the above Production Examples and a moisturizer will be described. In addition, all compounding amounts in the examples are parts by weight.

Fuji Oligo # 450 and Fuji Oligo G67 used as moisturizers in the following Examples are manufactured by Nippon Shokuhin Kako Co., Ltd., and Fuji Oligo # 450 is an oligosaccharide product containing about 50% by weight of maltotetraose. Fuji oligo G67 is an oligosaccharide product containing a total of about 40% by weight of maltohexaose and maltoheptaose. Similarly, Metroses 90SH-100 is hydroxypropylmethylcellulose manufactured by Shin-Etsu Chemical Co., Ltd., and 1.4% of the hydroxyl group in the glucose ring unit of cellulose is used.
This is a cellulose derivative in which 0.2 mol of hydroxypropoxyl groups are added per glucose ring unit of cellulose, each of which is substituted with a methoxyl group. DK ester SS
Is a mono-fatty acid ester of sucrose manufactured by Dai-ichi Kogyo Seiyaku Co., Ltd. As for polyvinyl alcohol, one having a degree of polymerization of about 500 was used.

[0045]

Examples 1 to 8 Wettable Powder The components shown in Table 1 were well mixed to obtain a wettable powder. Similarly, a wettable powder of Comparative Example containing spores of a bacterium belonging to the genus Bacillus but not containing a moisturizer was produced.

[0046]

[Table 1]

[0047]

[Examples 9 to 12] Wettable powder The spore fraction obtained in the above Production Example 2 and the synthetic hydrous silicon oxide powder were mixed at a weight ratio of 2: 1 to adjust the water content, and the water content was 6% by weight. A mixture of the spore fraction and the hydrous silicon oxide powder was obtained. Further, Fuji Oligo G67 and the synthetic hydrous silicon oxide powder were mixed at a weight ratio of 2: 1 to adjust the water content to obtain a mixture of Fuji Oligo G67 and the synthetic hydrous silicon oxide powder having a water content of 6% by weight. The components shown in Table 2 including these were well mixed to obtain a wettable powder. Similarly, a wettable powder of Comparative Example containing spores of a bacterium belonging to the genus Bacillus but not containing a moisturizer was produced.

[0048]

[Table 2]

[0049]

Example 13 Suspending Agent The components shown in Table 3 were thoroughly mixed to obtain a suspending agent. Similarly, a suspension of Comparative Example containing spores of a bacterium belonging to the genus Bacillus but not containing a moisturizer was produced.

[0050]

[Table 3]

<Evaluation of Agricultural and Horticultural Fungicide Composition of the Present Invention>
The plant disease control effect of the agricultural and horticultural fungicidal compositions obtained in each of the above Examples was evaluated together with the formulation of Comparative Example using cultivated plants such as cucumber, chrysanthemum, cyclamen, green beans, strawberries and eggplants.

(1) Gray mold control test of cucumber Cucumber (Hikari No. 3 P type) was sown in a plastic pot (diameter 6 cm) filled with soil and grown in a greenhouse for 12 to 14 days. A cucumber seedling with cotyledon expanded to a 500-fold diluted solution of the wettable powder obtained in Example 1 (4.0 × 10 ¹¹ CF).
(U / L) was spray-treated so as to adhere sufficiently to the leaf surface. After the treatment, the leaf surface was air-dried, and light (25 ° C, 12 hours), dark (1
It was left under dry conditions for 1 to 2 days. Cucumber cotyledons were cut from the hypocotyl, and an agar plug (diameter 6 mm) containing cucumber gray mold fungal spores was inoculated in the center of the cotyledon.
After inoculation, leave at 18 ° C, darkness and high humidity for 3 days,
The lesion size was measured. Further, a 500-fold diluted solution of the wettable powder obtained in Examples 2, 3, 6 to 10 (4.0 × 10 ¹¹ CF
U / L) was used to perform the same test.

Further, the cucumber cotyledons obtained in each of the above-mentioned examples which were not treated with the wettable powder were also subjected to the same test to measure the lesion size, and the control value of the test example was determined by the following formula using this as a control. Was calculated.

[0054]

[Equation 1] Control value (%) = (1- (lesion diameter of test example / lesion diameter of control)) × 100

For comparison, 5 of the wettable powder of Example 1 above was used.
The same test was performed using a 500-fold diluted liquid (4.0 × 10 ¹¹ CFU / L) of the wettable powder obtained in Comparative Examples 4 and 5 instead of the 00-fold diluted liquid, and measurement of lesion size and The control value was calculated. The results are shown in Table 4.

[0056]

[Table 4]

(2) Cucumber Powdery Mildew Control Test The control effect against cucumber powdery mildew was tested in a greenhouse (variety: Sagami Hanshibushiri, test section: 1 strain /
Section, 4 repetitions). The attack was spontaneous. From the true leaf stage of cucumber, a 200-fold dilution of the wettable powder obtained in Example 9 (1.0 × 10 ¹² CFU / L) was sprayed twice at 7-day intervals (spraying treatment on the whole strain ( Foliage spray), 50mL /
7 days after the final spraying, the disease degree was investigated for 5 true leaves from the lowest leaves. In addition, a 200-fold diluted solution of the wettable powder obtained in Example 11 (1.0 × 10 ¹² CFU / L)
The same test was performed using.

A similar test was conducted on cucumbers not treated with the wettable powder obtained in each of the above-mentioned examples, and the degree of disease was investigated. Using this as a control, the control value of the above-mentioned test examples was determined. Was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 9 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / L), a similar test was conducted using a commercially available morestan wettable powder (main component; quinomethionate, 333 ppm manufactured by Mikasa Chemicals),
The disease severity was investigated and the control value was calculated. The results are shown in Table 5.

[0060]

[Table 5]

(3) Control test against chrysanthemum white rust The control effect against chrysanthemum white rust was tested in a greenhouse (variety: Hideyoshi's power, test group: 10 strains / group, 3 repetitions). The chrysanthemum rust fungus was inoculated by arranging planters planting chrysanthemum infected with the white rust fungus in the test plot. During the middle stage of growth of chrysanthemum, 7 times the 200-fold dilution of the wettable powder obtained in Example 9 (1.0 × 10 ¹² CFU / L) was used.
Spray 3 times at daily intervals (spraying the entire plant (spraying)
(100 mL / strain), and the diseased leaf rate was investigated 7 days after the final spraying. A similar test was conducted using a 200-fold diluted liquid (1.0 × 10 ¹² CFU / L) of the wettable powder obtained in Example 12.

A similar test was conducted on chrysanthemums that were not treated with the wettable powder obtained in each of the above Examples, and the diseased leaf ratio was calculated. The control value of the test example was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 9 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / L), a similar test was performed using a commercially available suprol emulsion (main component; triphorin, manufactured by Takeda Pharmaceutical Co., Ltd., 1000 ppm), and the diseased leaf rate was investigated and the control value was calculated. The results are shown in Table 6.

[0064]

[Table 6]

(4) Cyclamen gray mold control test The control effect against cyclamen gray mold was tested in a greenhouse (variety: bolero, test section: 5 pots / section). At the flowering stage of cyclamen, 200 of the suspension obtained in Example 13
A doubling dilution (1.0 × 10 ¹² CFU / L) was sprayed (20 mL / strain) on the entire pot. The cyclamen Botrytis cinerea was inoculated by spraying the Botrytis cinerea spore suspension (1.0 × 10 ⁹ cells / L) onto the entire pot 7 days after the suspension treatment. The management was carried out under humid conditions by covering each pot with a plastic bag. Seven days after inoculation of the pathogen, the degree of disease in the petals (the disease was found only in the petals) was investigated.

The same test was carried out on cyclamen not treated with the suspending agent obtained in Example 13 above, and the degree of disease was calculated. Using this as a control, the test for cyclamen was carried out in the same manner as in the above-mentioned cucumber test example. The control value of the example was calculated.

Further, for comparison, instead of the 200-fold dilution of the suspension of Example 13 above, a 200-fold dilution of the suspension obtained in Comparative Example 6 (1.0 × 10 ¹² CFU / The same test was carried out using L) to investigate the degree of disease and calculate the control value. The results are shown in Table 7.

[0068]

[Table 7]

(5) Kidney Kernel Disease Control Test The control effect against kidney bean kernel disease was tested (variety: Satsuki Midori No. 2, test area: 6 m ² / area, no repetition). The attack was spontaneous. At the flowering stage of green beans, a 200-fold diluted solution of the wettable powder obtained in Example 2 (1.0 × 10 ¹² CF
(U / L) was sprayed 4 times at a rate of 300 L / 10a at 7-day intervals (spraying treatment (spraying on foliage) over the entire plant). Two weeks after the final spraying, the kidney pods were examined for the presence of disease.

The same test was carried out on the green beans obtained in Example 2 which had not been treated with the wettable powder, and the diseased pod rate was calculated. The control value of the test example was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 2 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / L), a similar test was carried out using a commercially available Loniran wettable powder (main component: vinclozolin, manufactured by Sankyo, 1000 ppm) to investigate the disease pod rate and calculate the control value. Table 8 shows the results.

[0072]

[Table 8]

(6) Strawberry Anthracnose Control Test The control effect on strawberry anthracnose was tested in the field (variety: Onamine, test section: 40 strains / section, 3 repetitions). The attack was spontaneous. At the seedling growing stage of strawberries, a 200-fold diluted solution of the wettable powder obtained in Example 3 (1.0 × 10 ¹² CFU /
L) was sprayed three times at intervals of 7 days at a rate of 400 L / 10a (spray treatment (spraying on foliage) over the whole plant). The diseased leaflet rate was investigated 7 days after the final application.

The same test was carried out on the strawberries that were not treated with the wettable powder obtained in Example 3 above, and the diseased leaflet rate was calculated. The control value of the test example was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 3 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / L), a similar test was performed using a commercially available anthracol wettable powder (main component; propineb, manufactured by Nippon Bayer Agrochem, 2000 ppm), and the diseased leaflet rate was investigated and the control value was calculated. The results are shown in Table 9.

[0076]

[Table 9]

(7) Gray mold disease control test of eggplants The control effect against eggplant gray mold disease was tested in a glass greenhouse (variety: 1,000 units, test section: 8 strains / section, 3 repetitions).
The inoculation of the eggplant Botrytis cinerea was carried out by suspending the cultivated eggplant fruits infected with Botrytis cinerea in a greenhouse. At the flowering and fruiting stage of the eggplant, a 200-fold diluted solution of the wettable powder obtained in Example 4 (1.0 × 10 ¹² CFU / L) was sprayed three times at 7-day intervals (spraying was applied to the entire plant (stems and leaves). Part spraying), 300L / 10
a) was done. 7 days after the final application, the diseased fruit / flower ratio was calculated. A similar test was conducted using the 200-fold diluted liquid of the wettable powder obtained in Example 5 (1.0 × 10 ¹¹ CFU / L).

The same test was carried out on eggplants not treated with the wettable powder obtained in each of the above-mentioned examples to calculate the disease-causing fruit and flower rate, which was used as a control in the same manner as in the above-mentioned cucumber test example. The control value of eggplant test examples was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 4 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / The same test was performed using L) to calculate the diseased fruit / flower ratio and the control value. The results are shown in Table 10.

[0080]

[Table 10]

(8) Gray mold disease control test of eggplant (sustainability stability test) The control effect against eggplant gray mold disease was tested in a glass greenhouse (variety: 1,000 units, test group: 8 strains / group, 3 repetitions).
The inoculation of the eggplant Botrytis cinerea was carried out by suspending the cultivated eggplant fruits infected with Botrytis cinerea in a greenhouse. At the flowering stage of eggplant, the 200-fold diluted solution (1 × 10 ¹² CFU / L) of the wettable powder obtained in Example 12 was sprayed 3 times at intervals of 7 days (spraying the entire strain, 300 L / 10a). did. The diseased fruit / flower ratio (%) was calculated 1 week and 2 weeks after the final application.

The same test was carried out on eggplants obtained in Example 12 which had not been treated with the wettable powder, and the diseased fruit / flower ratio was calculated. Using this as a control, the same procedure as in the above-mentioned cucumber test example was used. The control value of eggplant test examples was calculated.

Further, for comparison, instead of the 200-fold dilution of the wettable powder of Example 12 above, a 200-fold dilution of the wettable powder obtained in Comparative Example 4 (1.0 × 10 ¹² CFU / L),
The same test was performed using the viable cell suspension obtained in Comparative Example 3 and the culture solution supernatant (antibiotic fraction) obtained in Comparative Example 1, and the calculation and control of diseased fruit / flower ratio Valuation was calculated.
The results are shown in Table 11.

[0084]

[Table 11]

As is clear from these results, the agricultural / horticultural fungicide composition of the present invention containing spores of a bacterium belonging to the genus Bacillus and a moisturizer contains spores of a bacterium belonging to the genus Bacillus, Excellent in disease control activity as compared to fungicide composition containing no agent, and the disease control activity is equal to or more than that of conventional synthetic pesticides which are likely to cause physiological disorders, and further, of bacteria belonging to the genus Bacillus. It has good fixability and long-term stability compared to live cells and antibiotics obtained from it.

[0086]

EFFECTS OF THE INVENTION The fungicide composition for agricultural and horticultural use of the present invention contains a spore of a bacterium belonging to the genus Bacillus and a moisturizer, and thus has an excellent disease control action, and further has good fixability and long-term stability. Further, by applying this fungicide composition for agricultural and horticultural use, it is possible to very effectively protect plants from various diseases.

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Claims (7)

[Claims] 1. A fungicide composition for agricultural and horticultural use containing a spore of a bacterium belonging to the genus Bacillus and a moisturizer. 2. The fungicide composition for agricultural and horticultural use according to claim 1, wherein the bacterium belonging to the genus Bacillus competes with a plant pathogen. 3. The fungicide composition for agricultural and horticultural use according to claim 2, wherein the bacterium belonging to the genus Bacillus that antagonizes the plant pathogen is Bacillus subtilis. 4. The Bacillus subtilis, the Bacillus subtilis, and the Bacillus subtilis FERM.
P-14647 strain, Bacillus subtilis FERM
The agricultural and horticultural fungicide composition according to claim 3, which is selected from P-14646 strains. 5. The agricultural and horticultural method according to claim 1, wherein the spore content of the bacterium belonging to the genus Bacillus is 0.001 to 99.9 parts by weight based on the total amount of the composition. Disinfectant composition. 6. The moisturizer according to claim 1, wherein the moisturizer is selected from polysaccharides, oligosaccharides, and synthetic water-soluble polymers.
The agricultural and horticultural sterilizing composition according to the item. 7. The content of the moisturizing agent is 0.1 to 99.999 parts by weight with respect to the total amount of the composition.
The fungicide composition for agricultural and horticultural use according to any one of 1.