JPH10316519A - Controlling agent against plant disease injury - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject controlling agent capable of effectively controlling plant disease injuries by including a cultured product of a microorganism, belonging to Clostridium butyricum and having antimicrobial actions on plant disease injury molds and plant disease injury bacteria as an active ingredient therein. SOLUTION: This controlling agent is obtained by including a cultured product of a microorganism belonging to Clostridium butyricum which is an anaerobe capable of manifesting antimicrobial actions on plant disease injury molds such as Fusarium oxysporum and plant disease injury bacteria such as Pseudomonas cichorii (e.g. Clostridium butyricum MIYAIRI 588 or Clostridium butyricum NIP1020) as an active ingredient in an amount of preferably 2-80 wt.% in the controlling agent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a plant disease controlling agent. More specifically, the present invention relates to a plant disease controlling agent using anaerobic bacteria used for plant diseases.

[0002]

2. Description of the Related Art Plants, such as useful crops and fruit trees, are of various diseases, whether annual or perennial. Some of the causes of this disease are soil conditions, weather conditions or mechanical damage (non-communicable), but infectious diseases, especially those caused by bacteria and molds, are contagious, In the worst case, crops may not be able to be harvested at all, so they are important as control targets. Such plant diseases include:
Due to filamentous fungi, for example, sweet potato, tomato, cucumber, cabbage, radish, strawberry, spinach, carnation, aster, cyclamen, margaret, alfalfa and other wilt diseases by Fusarium oxysporum, verticillium Eggplant, strawberry, chrysanthemum, chrysanthemum, chrysanthemum, tomato, okra, butterbur, watermelon, melon, cucumber, capsicum and potato, etc. by dariae (Verticillium dahliae), yellowing of Chinese cabbage, Rhizoctonia solani (Rhizocton)
ia solani) flax, sumac, peas, cabbage,
Buckwheat, tobacco (short sickness), onion, dahlia, tuna,
Sugar beet, capsicum, tomato, eggplant, carrot, cotton, soybean, cucumber, radish, wheat, green onion, goby, spinach, makuwauri, honeysuckle, seedling wilt of yuugao and cotton (knee-fold disease), Roselinia necatrix ( Rosellinia necatrix) causes white root rot and Helicobasidium monpa (Helicob) in 60 plants of 34 families, including pear, tangerine, tea, mulberry, apple and fir.
asidium mompa) 50 departments excluding grasses such as sweet potato, mulberry, apple, asparagus and alfalfa 11
Purple root rot of nine plants, and those caused by bacteria, for example, soft rot such as lettuce caused by Pseudomonas cichorii, Erwinia carotobola (E
(rwinia carotovora) by Chinese cabbage, cabbage, potatoes, various vegetables including onions and konjac, and various varieties of plants such as irises, irises and cyclamen and various flowering plants, such as soft rot and Xanthomonas campestris p. campestris ( Xanth
omonas campestris pv. campestris), there are various diseases caused by various plant pathogenic fungi such as cabbage black rot.

[0003] Conventionally, with respect to the control of such plant diseases using microorganisms, Bacillus spp.
Aerobic bacteria such as illus, Pseudomonas, Agrobacterium and lactic acid bacteria have been mainly used as antagonistic microorganisms. If a disease control effect is to be exerted using such a bacterium, it must first have an antagonistic action against the disease bacterium, and if the bacterium itself is used as a plant disease control agent, It is necessary to establish conditions that allow the cells to proliferate sufficiently at the intended location and exhibit the intended function.

However, in soil, the upper surface is quite aerobic, but after the growth of aerobic bacteria, the lower part of the soil or the center of individual soil aggregates is anaerobic, There are also many bacterium. In addition, oxygen in the soil is present only in an amount that can be consumed by plant roots and microorganisms in the soil per day. Oxygen is quickly consumed and becomes anaerobic. Therefore, complete control of plant pathogenic fungi using only aerobic bacteria can be applied favorably to shallow soil with oxygen or aerobic soil with many soil gaps. At present, it is often difficult to control disease with aerobic bacteria alone in soils under anaerobic conditions such as soils in deep strata.

[0005] In addition to the above case, the reduced layer portion of the desalinated paddy soil is also anaerobic, and oxygen is also present in the central part of the water-saturated soil aggregate having a diameter of 3 mm or more in the field soil. It is said that there is no. Furthermore, even in other soils, after aerobic bacteria grow, they generally become anaerobic, and the growth of anaerobic bacteria becomes active.

[0006] As described above, the soil in which the plant disease bacteria inhabit is not always in an aerobic state, and often cannot be prevented only by a conventional plant disease controlling agent mainly composed of aerobic bacteria. is there.

[0007] Microorganisms contained in currently used plant disease control agents are not necessarily known to be toxic to humans, and there remains a problem in terms of safety for human beings.

[0008]

Accordingly, an object of the present invention is to provide a plant disease controlling agent which can effectively control plant diseases.

[0009] Another object of the present invention is to effectively and safely control the disease caused by a plant disease bacterium by a highly safe anaerobic bacterium having an antagonistic action against a plant disease bacterium.

[0010] The present invention furthermore effectively and safely controls plant disease caused by infection with plant disease molds including Fusarium oxysporum and plant disease bacteria including Pseudomonas cichorii. The purpose is to do so.

[0011]

In view of such circumstances, the present inventors have proposed the use of an anaerobic bacterium that can grow in a deep rhizosphere flora and the like that cannot be grown by an aerobic bacterium. They have discovered that the effects can be exerted, and based on this finding, have completed the present invention.

That is, the above objects are achieved by the following (1) to (8).

(1) Clostridium butyricum (Clo
A plant disease control agent comprising, as an active ingredient, a culture of a microorganism which belongs to S. stridium butyricum) and has an antibacterial action against plant disease fungi and plant disease bacteria.

(2) The plant disease-causing fungus is Fusarium
The plant disease controlling agent according to the above (1), which is oxysporum (Fusarium oxysporum).

(3) The plant disease controlling agent according to (1) or (2), wherein the plant disease bacterium is Pseudomonas cichorii.

(4) The microorganism is Clostridium butyricum M. 588 (Clostridium butyricum M.
IYAIRI 588), Clostridium, Butyricum, NIP
1020 (Clostridium butyricum NIP1020), Clostridium butyricum NIP1021 (Clostridium byricum NIP1020)
utyricum NIP1021), Clostridium butyricum
ATCC860 (Clostridium butyricum ATCC860) or Clostridium butyricum ATCC19398
(Clostridium butyricum ATCC19398), the plant disease control agent according to any one of the above (1) to (3).

(5) The microorganism is Clostridium butyricum M. 588 (Clostridium butyricum M.
IYAIRI 588). The plant disease control agent according to (4) above.

(6) A plant disease controlling agent obtained by mixing a culture of the microorganism according to any one of (1) to (5) above with a carrier.

(7) A plant disease controlling agent obtained by mixing a culture of the microorganism according to any one of (1) to (5) with other microorganisms.

(8) The plant disease controlling agent according to the above (7), wherein the other microorganism is yeast.

[0021]

BEST MODE FOR CARRYING OUT THE INVENTION The plant disease controlling agent of the present invention is a plant disease fungus such as Fusarium oxysporum and Pseudomonas chicory.
Clostridium butyricum (Clostr. cichorii)
idium butyricum) as an active ingredient.

Hereinafter, the present invention will be described in detail.

The microorganism used in the plant disease control agent of the present invention is Clostridium butyricum.
um butyricum) and belongs to Fusarium oxysporum (F
(Usarium oxysporum) and other microorganisms that exhibit an antibacterial action against plant disease filamentous fungi such as Pseudomonas chicory (Pseudomonascichorii), but are not particularly limited.Specifically, Clostridium butyricum Miyairi 588 ( Clostridium butyricum MIYAIRI
588), Clostridium butyricum NIP102
0 (Clostridium butyricum NIP1020) (Accession number: FE
RM BP-5794), Clostridium butyricum NIP1021
(Accession number: FERM BP-5795), Clostridium butyricum ATCC 860 (Clostridium but
yricum ATCC860) and Clostridium butyricum.
ATCC 19398 (Clostridium butyricum ATCC 1939
8) and the like. Of these, preferably Clostridium butyricum Miyairi 588 (Clostridi
um butyricum MIYAIRI 588) (hereinafter simply referred to as “588 strains”
The microorganism has been deposited with the Ministry of International Trade and Industry at the National Institute of Advanced Industrial Science and Technology, under the accession number of MIC No. 2789 (FERM BP-2789).

In the present specification, the term "culture" includes a culture solution obtained by culturing the microorganism, a residue containing the microorganism obtained by centrifuging or filtering the culture solution, and a dried product of the residue.

In the present invention, the microorganism is cultured by a known culture method.
The case where eight strains are used is described below. 588 strains were transformed from 1.0 (w / v)% peptone, 1.0 (w / v)%
A medium comprising yeast extract, 1.0 (w / v)% corn starch and 0.2 (w / v)% precipitated calcium carbonate has a concentration of 10 ⁴ to 10 ⁷ cells / ml, preferably 10 ⁵ to 10 ⁶ cells / ml. And inoculated at 30-40 ° C for 12-72.
Incubate for static time. Next, the culture is centrifuged (2,000 to 8,000 g × 10 to 30 minutes) or filtered to obtain a residue of 588 strains. Further, the residue is subjected to a drying treatment by air drying or the like or a reduced pressure drying treatment at 0 to 60 ° C. for 1 to 20 hours to obtain a dried product of the residue.
In each of the above steps, a culture of the target microorganism is obtained as a culture solution, a residue, and a dried product of the target microorganism.

The plant disease controlling agent of the present invention may consist of a culture of the above microorganism alone or may be a mixture of a culture of the above microorganism with a culture of another microorganism. As the other microorganisms when the plant disease controlling agent of the present invention comprises a mixture of a culture of a microorganism and a culture of another microorganism, generally used non-pathogenic ones can be used. As Bacillus subtilis (B
acillus subtilis), Bacillus thurigensis (Bac
illus thuringiensis) and Bacillus cereus (Bacillus)
cereus), Bacillus, Pseudomonas
Pseudomonas fluorescens, Pseudomonas gladioli, Pseudomonas cepacia, Pseudomonas glumae, Pseudomonas avenae (Pseudomonas avenae) Agrobacterium (Agrobacterium) such as Radiobacter (Agrobacterium radiobacter), Enterobacter (Enterobacter) such as Enterobacter aglomerans, Xanthomonas (Xanthomonas)
, Bacteria such as Pasteuria, lactic acid bacteria (Lactobacillus), Gliocladium, Trichoder
ma), filamentous fungi such as genus Aspergillus (Aspergillus), genus Penicillium, genus Rhizopus, genus Fusarium (Fusarium), and yeasts (for example, Saccharomyces and Candida)
), Sporidesmium and VA mycorrhizal fungi. Of these, yeast is particularly preferably used. In the present invention, the culture of the other microorganism may be used alone or in combination with the culture of the other microorganism.

In the present invention, the cultivation of the other microorganism when the above-mentioned other microorganism is used depends on the kind of the microorganism used, and is performed by a known culturing method.

The medium used for culturing the microorganism or other microorganisms according to the present invention varies depending on the kind of the strain used, etc., but the carbon source, the appropriate amount of nitrogen source, the inorganic salt and the vitamin which the microorganism used can utilize. The medium may be any synthetic or natural medium, as long as it contains other nutrients such as microorganisms and is capable of growing the microorganism according to the present invention or other microorganisms.

For example, examples of the carbon source used in the culture according to the present invention are not particularly limited as long as the strain used can be assimilated. The carbon source is not necessarily limited to sugar, but in consideration of the growth of the strain and the like, a sugar or a sugar containing sugar that can be used by the strain to be used is preferably used. Specifically, cellobiose, glucose, fructose, galactose, lactose, maltose, mannose, melibiose, raffinose,
Salicin, starch, sucrose, trehalose, xylose, dextrin, molasses and the like. Among them, starch, glucose, fructose, sucrose, molasses, lactose and the like are preferably used. One or more of the above carbon sources may be selected and used in consideration of the assimilation of the microorganism used. At this time, the addition concentration of the carbon source varies depending on the strain to be used, the type of the carbon source, the composition of the medium to be used, and the like, but is usually 0.5 to 5.0 (w / v)%, preferably 1 to 5.0 (w / v)%. 3
(W / v)%.

As the nitrogen source used in the culture according to the present invention, a nitrogen source generally used for culturing microorganisms is used. Examples thereof include meat extract, polypeptone, peptone, yeast extract and taste liquid. Soybean and wheat hydrolysates, soybean powder, milk casein, casamino acids, various amino acids, corn steep liquor, other animals, etc.
Organic nitrogen compounds such as hydrolysates of plants and microorganisms, and ammonium salts such as ammonium sulfate.
Among these nitrogen sources, peptone, yeast extract, meat extract, corn steep liquor, taste liquid, casamino acid and the like are preferably used. One or more of the above-mentioned nitrogen sources may be selected and used in consideration of the growth of the microorganism to be used. At this time, the addition concentration of the nitrogen source varies depending on the strain used, the type of the nitrogen source, the composition of the medium used, and the like, but is usually 0.5 to 4.0 (w / v)%, preferably 2 to 4.0 (w / v)%. 3 (w / v)%.

Further, as the inorganic salt used in the culture according to the present invention, those known to those skilled in the art are usually used, for example, magnesium, manganese, calcium, sodium, potassium, molybdenum, strontium, boron, copper, iron, One or more selected from phosphate, hydrochloride, sulfate, butyrate, propionate, acetate and the like of tin and zinc can be used. In the medium, if necessary, an antifoaming agent, a vegetable oil, a surfactant,
Blood and blood components, drugs such as vitamins and antibiotics, physiologically active substances such as plant or animal hormones and the like may be appropriately added.

In the present invention, the culturing conditions of the microorganisms vary depending on the kind of microorganisms to be used and the like. However, Clostridium butyricum is obligately anaerobic, so it is not ventilated or ventilated with nitrogen or carbon dioxide gas. While cultivating under anaerobic conditions while lowering the oxidation-reduction potential by adding a reducing agent to the culture medium, or when using aerobic microorganisms, agitation is performed while stirring or aerating oxygen. And culturing under appropriate conditions. Culture conditions in the present invention are appropriately selected depending on the range of growth of the strain to be used, the composition of the medium, and the culture method (static or shaking culture), and are known to those skilled in the art.
Specifically, when using the microorganism according to the present invention, the culture temperature is usually 30 to 40 ° C, preferably 35 to 37 ° C.
C. and the pH of the medium is usually 5 to 8, preferably 6
When using other microorganisms according to the present invention, the culture temperature is usually 20 to 40 ° C., preferably 25 to 40 ° C.
~ 35 ° C, and the pH of the medium is usually 5-8, preferably 6-7.

Alternatively, the plant disease controlling agent of the present invention may be obtained by mixing a culture of the above microorganism with a carrier. As the carrier used when the plant disease controlling agent of the present invention comprises a mixture of a microorganism culture and a carrier, those generally used as carriers can be used. Specifically, vermiculite, zeolite, diatomaceous earth, tankal, rough tankal, silicate, limestone, volcanic ash, red soil,
Activated silicic acid, metal oxide, porous mineral, soft ceramics, sulfur precision residue, humus, animal lime, or fish cake, dried fish, fish boiled meal, meat meal powder, meat-and-bone meal, raw bone meal,
Steamed bone meal, dry blood (blood meal), earthworm dung, or rapeseed oil residue, soybean oil residue, cottonseed oil residue, peanut oil residue,
Castor oil grounds, kabok oil grounds, linseed oil grounds, sesame grounds, rice bran oil grounds, food and brewed grounds (soy sauce grounds powder, amino acid by-product fertilizer, fermentation by-product fertilizer), compost, cow manure, pig manure, chicken manure, human Manure, green manure, crab chaff, giant clam chaff, shell powder, shell fossil powder, peat (beet) (processed peat, sphagnum, humic acid material), chaff, sawdust, charcoal and charcoal. Of these, vermiculite, zeolite, limestone, chicken dung and sawdust are preferably used. The carrier may be in solid form or in liquid form.

The plant disease controlling agent of the present invention may contain, in addition to the above-mentioned microorganisms and / or other microorganisms or carriers, antibacterial substances such as wood vinegar, soil disinfectants, antibiotics, chemical fertilizers, and disease control as required. It may contain soil and the like.

When the culture of the microorganism is used alone in the plant disease control agent of the present invention, the abundance of the culture of the microorganism is 2 parts relative to the total components of the plant disease control agent.
~ 80 (w / w)%, preferably 2 ~ 30 (w / w)%
It is. When a culture of the above microorganism is used in combination with a culture of another microorganism, the total abundance thereof is 2 to 80 with respect to the total plant disease controlling component.
(W / w)%, preferably 40-60 (w / w)%, and the mixing ratio of the culture of the microorganism to the culture of the other microorganism (weight of the culture of the microorganism: weight of the culture of the other microorganism) Weight) is 1: 5 to 5: 1, preferably 1: 2 to 2: 1. Furthermore, when a culture of the microorganism is used in combination with a carrier, the total amount of these is 2 to 80 (w / w)%, preferably 2%, based on the total plant disease controlling agent components. And the mixing ratio of the culture of the microorganism to the carrier (weight of the culture of the microorganism: weight of the carrier) is 1: 5 to 5: 1, preferably 1: 2 to 2: 1
It is.

The plant disease controlling agent thus obtained can be applied to the microorganism of the present invention by spraying the culture of the microorganism of the present invention and / or the culture or carrier of another microorganism on the soil as it is. And / or other microorganisms or cultures or carriers in the form of a solution which is dispersed in water or a suitable solvent and sprayed, the culture of microorganisms and / or other microorganisms according to the present invention may be used. It has a form of wettable powder that mixes a culture or a carrier with a clay mineral carrier, a surfactant, a fixing agent, etc., grinds and mixes as a fine powder with a large amount of water before use and sprays it as a suspension. Or a mixture of a microorganism culture and / or other microorganism culture or carrier according to the present invention with a clay mineral or carbonated lime or the like, which is then sprayed as a fine powder. Is obtained by mixing and crushing a culture of a microorganism according to the present invention and / or a culture or carrier of another microorganism with a carrier of a clay mineral, adding a solidifying agent such as starch, a stabilizer and a surfactant as an auxiliary agent to obtain a granulated product. It has a well-known form such as a form of granules molded into a granule.

The amount of the plant disease control agent of the present invention applied to soil varies depending on the kind of pathogenic bacteria to be controlled and the degree of soil disease caused by the pathogenic bacteria, but is usually 0.1 to 2 kg in terms of microorganisms according to the present invention. / M ² , preferably 0.3 to
It is 1 kg / m ² .

Further, Clostridium butyricum used in the present invention is widely marketed as a probiotic agent, a feed additive, and a food, and is administered to mammals such as humans and domestic animals for a long period of time. No side effects are observed and high safety is guaranteed. Therefore, this Clostridium butyricum (C
The plant disease controlling agent of the present invention containing a culture of (Lostridium butyricum) as an active ingredient has high safety.

[0039]

The present invention will be described more specifically below with reference to examples. However, the present invention is not limited to these examples.

Example 1 Meat extract agar medium [meat extract 1.0%, peptone 1.0
%, Sodium chloride 0.5%, L-cysteine hydrochloride
Monohydrate 0.03%, glucose 0.5%, agar 1.5
%], Coat each of Clostridium butyricum shown in Table 1 with
Anaerobic culture was performed in an anaerobic glove box at 7 ° C for 48 to 96 hours. Separately from the above culture, each of the test pathogenic bacteria shown in Table 1 was smeared on the same meat extract agar medium as above, and cultured at 25 to 28 ° C. for 48 to 120 hours. Next, the suspension of the test microorganism cultured in this manner was added at a concentration of 10 ⁵ to 10 ⁶ cells / ml to a meat extract medium containing 0.8% agar and kept at 40 to 50 ° C. These were prepared and layered on an agar medium of Clostridium butyricum anaerobically cultured as described above. Furthermore, this overlay medium was cultured at 25 to 28 ° C for 24 to 9 days.
Aerobic culture was performed for 6 hours, and the presence or absence of the inhibition zone and its size were observed. Various Clostridium butyricum (Clostridium)
dium butyricum) was measured. The results are shown in Table 1 below. In Table 1, M588 indicates Clostridium butyricum Miyairi 588 (Clostridium b.
utyricum MIYAIRI 588) strain, NIP1020 is Clostridium, Butyricum, NIP1020 (Clostridium
butyricum NIP1020) strain and NIP1021 is Clostridium butilicum NIP1021 (Clostridium b.
utyricum NIP1021) strain and ATCC 860 is Clostridium butyricum ATCC 860 (Clostridium but
yricum ATCC860) strain and ATCC 19398 are Clostridium butyricum ATCC 19398 (Clo
stridium butyricum ATCC19398).

[0041]

[Table 1]

From the results shown in Table 1, it was found that the microorganism according to the present invention exerts a plant disease control effect on various plant disease fungi and plant disease bacteria. Also,
Among the microorganisms used, it was also shown that 588 strains exhibited a remarkable plant disease control effect against various plant disease fungi.

[0043]

As described above, the plant disease control agent of the present invention is Clostridium butilicum.
utyricum) and contains, as an active ingredient, a culture of a microorganism exhibiting an antibacterial action against plant-disease fungi and plant-disease bacteria. Therefore,
According to the present invention, even under anaerobic conditions, various plant disease fungi and plant disease bacteria, particularly Fusarium oxysporum (F
usarium oxysporum) and Pseudomonas chicory (Pseud
omonas cichorii), a plant disease controlling agent having an inhibitory effect on growth and effectively preventing and treating plant diseases of flowers and fruit trees, and furthermore, a plant as a microbial material centering on conventional aerobic bacteria A plant disease control effect on pathogenic bacteria on heavy clay soil and deep-layer soil, which is ineffective with the disease control agent, can be expected.

The microorganism belonging to Clostridium butyricum used in the present invention is administered as a useful bacterium to humans, livestock, poultry, and aquatic animals. Since the microorganism is a microorganism guaranteed to have extremely high safety, the agent for controlling plant diseases of the present invention can effectively control plant diseases and has high safety.

Furthermore, although the microorganism belonging to Clostridium butyricum used in the present invention is an anaerobic bacterium, it can survive without forming spores and dying even under aerobic conditions. The plant disease controlling agent can effectively control plant diseases under anaerobic conditions as well as under anaerobic conditions.

Claims (8)

[Claims] 1. Clostridium butyricum
and a plant disease control agent comprising, as an active ingredient, a culture of a microorganism belonging to the group consisting of P. dium butyricum) and having an antibacterial action against plant disease-causing fungi and plant disease bacteria. 2. The plant disease controlling agent according to claim 1, wherein the plant disease filamentous fungus is Fusarium oxysporum. 3. The plant disease controlling agent according to claim 1, wherein the plant disease bacterium is Pseudomonas cichorii. 4. The microorganism is Clostridium butyricum MIYAIRI 588.
588), Clostridium butyricum NIP102
0 (Clostridium butyricum NIP1020), Clostridium butyricum NIP1021 (Clostridium butyricum NIP1020)
um NIP1021), Clostridium, Butyricum, ATC
C860 (Clostridium butyricum ATCC860) or Clostridium butyricum ATCC19398 (Clost
ridium butyricum ATCC19398), the plant disease controlling agent according to any one of claims 1 to 3. 5. The method according to claim 5, wherein the microorganism is Clostridium butyricum MIYAIRI 588.
The plant disease controlling agent according to claim 4, which is 588). 6. A plant disease controlling agent obtained by mixing a culture of the microorganism according to claim 1 with a carrier. 7. A plant disease controlling agent obtained by mixing a culture of the microorganism according to any one of claims 1 to 5 with another microorganism. 8. The plant disease controlling agent according to claim 7, wherein the other microorganism is yeast.