JP4670254B2 - How to control harmful plant parasitic nematodes - Google Patents

Description

  The present invention relates to a method for controlling harmful plant parasitic nematodes and the like.

  Harmful plant parasitic nematodes refer to nematodes that grow while relying on nutrients in higher plants, and these parasitize crops and horticultural crops that are important for human life. Its parasitic range is so wide that it affects almost all higher plants, causing serious agricultural damage all over the world.

Currently, the removal and control of damage caused by harmful plant parasitic nematodes in the agricultural field is generally mainly controlled by chemical pesticides such as chloropicrin, methyl bromide, dazomet, aldicarb, oxamyl, and phosthiazate. Has been. However, these chemical pesticides alone may not always provide sufficient nematode control, and the development of other effective nematode control methods that can complement or substitute for chemical pesticides in nematode control may not be possible. It was sought after.
As one candidate technique for meeting such a demand, it is known that amino acids such as methionine suppress crop damage caused by harmful plant parasitic nematodes (see, for example, Non-Patent Document 1). .
On the other hand, as another candidate technique, a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria ATCC 46474 strain, which is a filamentous fungus, is effective against harmful plant parasitic nematodes. It is known to show a control effect (for example, refer to Patent Document 1).
US Pat. No. 5,051,255 Nematologica, Vol.17, pp.495-500 (1971), Plant Protection Vol.56, No.5 pp.22-25 (2001)

  However, the use of only amino acids such as methionine or the use of only the above-mentioned cultures may not always have the above-mentioned control efficacy.

  As a result of intensive studies under such circumstances, the present inventors have found that amino acids can be used in combination with a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof. The present inventors have found that this is an extremely effective means for controlling harmful plant parasitic nematodes, and have reached the present invention.

That is, the present invention
1. An effective amount of an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof is used to protect crops, harmful plant parasitic nematodes or harmful A method for controlling harmful plant parasitic nematodes, which is applied to a habitat of plant parasitic nematodes (hereinafter sometimes referred to as the method of the present invention);
2. A synergistic effective amount of
Amino acids,
A culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof,
A method for controlling harmful plant parasitic nematodes characterized by being applied to crops to be protected, harmful plant parasitic nematodes or habitats of harmful plant parasitic nematodes;
3. 3. The method according to item 1 or 2 above, wherein the amino acid is methionine;
4). 3. The method of item 1 or 2 above, wherein the amino acid is applied from about 0.05 kg to about 50 kg per 1000 m ² ;
5. 3. The method according to item 1 or 2 above, wherein the Myrothecium verrucaria is Myrothecium verrucaria ATCC 46474 strain;
6). A harmful plant parasitic nematode control agent characterized by comprising an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof (hereinafter, Sometimes referred to as the present control agent).
7). A synergistic effective amount of
A harmful plant parasitic nematode control agent comprising an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof;
8). The harmful plant parasitic nematode controlling agent according to the above 6 or 7, wherein the amino acid is methionine;
9. The harmful plant parasitic nematode control agent according to 6 or 7 above, wherein Myrothecium verrucaria is Myrothecium verrucaria ATCC 46474 strain;
10. Use of a combination of an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof for controlling harmful plant parasitic nematodes;
11. A synergistic amount for the control of harmful plant parasitic nematodes,
Use of a combination of an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof;
12 12. The use according to item 10 or 11 above, wherein the amino acid is methionine, alanine, leucine, phenylalanine, valine, aspartic acid, aminobutyric acid or ethionine, or a mixture of all or part thereof;
13. 12. The use according to the above 10 or 11, wherein the amino acid is methionine;
Etc. are provided.

  INDUSTRIAL APPLICABILITY The present invention can provide an effective nematode control method and the like that can supplement or replace chemical pesticides in nematode control.

  In the present invention, amino acids used as an active ingredient for controlling harmful plant parasitic nematodes include, for example, natural amino acids such as methionine, alanine, leucine, phenylalanine and valine, aspartic acid, aminobutyric acid, and ethionine. Such artificial amino acids as well as a mixture of all or a part thereof can be mentioned. Preferable examples include methionine, phenylalanine, and valine.

As the “culture having anti-nematode activity obtained by culturing Myrothecium verrucaria” in the present invention, for example, M. verrucaria is used under the following culture conditions. Examples thereof include a culture having anti-nematode activity obtained by culturing (for example, a whole medium containing mycelium after culturing). In addition, the “equivalent of a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria” in the present invention includes, for example, a sterilized product of the above-mentioned culture, or a product derived from the sterilized product. An extract, a partially purified product or a purified product; or an extract, a partially purified product or a purified product from the above culture, or a sterilized product thereof, which has anti-nematode activity. Here, examples of the solvent used for extraction include acetone, methanol, and the like. Partial purification or purification can be performed by appropriately combining ordinary fractionation and purification methods including centrifugation, filtration, extraction and the like. Examples of the sterilization method include conventional methods such as heat sterilization, filter sterilization, alcohol sterilization, and gas sterilization, and preferably heat sterilization.
Representative of the above-mentioned “culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof” (hereinafter sometimes referred to as the main culture). As an example, a main culture (see US Pat. No. 5,051,255) obtained by culturing Milotesium velukaria ATCC 46474 can be mentioned.

Here, “Myrothecium verrucaria” is, for example, a filamentous fungus having the following characteristics.
(1) It belongs to Hyphomycetes and does not form conidia shells (pycnidia) or conidia layers (acervuli).
(2) Conidiophore is colorless, has partition walls, and is glassy.
(3) Conidia are spindle-shaped, single vesicle, dark olive to dark green, and have a size of 2.0 μm to 3.0 μm × 7.0 μm to 8.0 μm.
(4) Growth is possible on a potato-glucose medium at a temperature of 8 ° C to 40 ° C, but the optimum temperature is about 25 ° C.
(5) The mycelium is colorless and has a thin partition wall, and the size is about 1.5 μm × 3.0 μm.

  A typical example of Myroteticum velukaria used in the present invention includes Myrothecium verrucaria ATCC 46474 strain. The strain is registered under the accession number ATCC 46474 in the permanent culture collection of the American Type Culture Collection (Parklawn Drive 12301, Rockville, Md. USA).

The main culture used in the present invention can be obtained and prepared by the following method, for example.
In order to obtain the main culture, Myrothecium verrucaria, which is a filamentous fungus such as Myrothecium verrucaria ATCC 46474, may be cultured in a nutrient medium.
The culture may be initiated from the spore of the microorganism or a specially grown inoculum. For example, spores produced on an initial growth medium such as potato-dextrose agar medium are transplanted into a growth medium (seed culture solution) in a flask and cultured while shaking. Thereby, spore germination and initial growth of the microorganism are performed. The optimum spore concentration used as the inoculation source can be easily determined by a normal routine preliminary test or the like.

  The germinated spores in an activated growth state may then be transplanted as a seed source to a fermenter containing a nutrient medium. In addition, what is necessary is just to prepare the inoculation source used as the amount of inoculation of 1-2% normally in the culture | cultivation for obtaining this culture. Here, the “fermenter” is not particularly limited, and various cultures such as a shaking culture method, a solid culture method, a continuous culture method, and a batch culture method, which are suitable from a laboratory level to a large-scale culture process. Figure 2 shows an apparatus used in the law.

The culture can be performed in a shake flask or a stationary fermenter.
When a shake flask is used, air is supplied by stirring the flask so that air is mixed into the medium. In order to stir the flask, a wide range of shaking culture apparatuses can be used. In the process, a rotary shaker or a reciprocating shaker is generally used. It is desirable to use a rotary shaker such that the flask rotates at about 100-500 rpm, preferably about 200-250 rpm and a circular trajectory of about 50 mm. The culture usually moves smoothly along the inside of a flask, such as an Erlenmeyer flask.
When using a stationary fermenter, air is supplied by injecting air or oxygen into the culture mixture by propulsion means such as a disc turbine, wing disc, open turbine or marine propeller.

The nutrient medium used for culturing the microorganism is not particularly limited as long as the microorganism grows, and is assimilated by a carbon source, a nitrogen source, an inorganic salt, and the microorganism normally used for culturing the microorganism. A medium appropriately containing such growth factors is used.
Preferable carbon sources used in the medium include saccharides such as dextrose, glucose, lactose and maltose, starch, dextrin, corn meal and glycerol.
Examples of the nitrogen source used in the medium include an organic nitrogen source, an inorganic nitrogen source, or a mixture thereof. Specifically, examples of the nitrogen source used in the medium include soy flour, Corn Steep Liquor, peanut meal, cottonseed meal, cotton germ meal (Corn Germ Meal), and various ammoniums. Examples include salts.
Furthermore, it is beneficial to add minerals, growth factors and the like to the medium. For example, Distillers' solubles, Corn Steep Liquor, Fish meal, yeast products, Crude medium ingredients such as peptone milk and whey It contains not only minerals but also growth factors.
As inorganic salts used in the liquid medium, for example, inorganic salts such as potassium phosphate, sodium chloride, ferrous sulfate, calcium carbonate, cobalt chloride, magnesium sulfate, and zinc sulfate can be added to the medium. A solid material such as calcium carbonate is preferably added to assist pH adjustment in the process (culture).
Preferred examples of the culture time for obtaining the main culture include about 4 days to about 7 days. More preferably, 5 days is mentioned. As a culture temperature, about 20-40 degreeC can be mentioned. Preferably, it is about 25 degreeC-about 30 degreeC, Most preferably, about 25 degreeC is mentioned. Examples of the medium pH include about 4 to about 10. Preferably, it is about 5 to about 8, and most preferably neutral pH.

  The main culture may be collected by a method usually used in this field.

  By extracting an anti-nematode active ingredient from the main culture thus prepared using an appropriate solvent such as acetone, the main culture as an equivalent can also be prepared. Furthermore, the main culture as a powder can also be prepared by freeze-drying such a main culture and then converting it to a powder state. Alternatively, a commercially produced main culture may be purchased and used. For example, a product of the main culture such as trade name: DiTera (manufactured by Valent Biosciences Corporation) is known, but is not limited thereto.

  Examples of harmful plant parasitic nematodes that can be controlled in the present invention include the following.

  Meloidogyne nematodes such as sweet potato nematodes (Meloidogyne incognita), Java root-knot nematodes (Meloidogyne javanica), northern root-knot nematodes (Meloidogyne hapla), arealite root-knot nematodes (Meloidogyne arenity, etc.) Ditylelenchus nematodes, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus cffeae, Pratylenchus vulnus, etc. Globodera nematodes such as cyst nematode (Globodera rostochiensis), potato white cyst nematode (Globodera pallida), soybean cyst nematode (Heterodera glycines), sugar beet cyst nematode (Heterodera shunt) Heterodera nematodes such as achtoii), Aphelenchus nematodes such as Aphelenchus avenae, Radopholus nematodes such as Radopholus similis, and the like are limited thereto. Is not to be done.

The method of the present invention is not particularly limited, but is not limited tomato, dungeon thistle, Jerusalem artichoke, eggplant, banana, barley, beet edible root, cacao, carrot, cassava, celery, chickpea, citrus, coconut, coffee, corn, persimmon , Cowpea, field bean, fodder, grapes, guava, melon, millet, oats, okra, ornamental plants, papaya, peanuts, pepper, yellow beans, pineapple, potato, rice, rye, sorghum, It can be used for nematode control for various agricultural applications in various different plants and fruits such as soybean, sugar beet, sugar cane, pepper, sweet potato, tea, tobacco, various lettuce, wheat and yam.
In addition, cultivated flowers such as carnations, rose trees, gerberas, chrysanthemums, potted plants, philodendrons, figs, pothos, chitoserains (sansevierias) and cactus can also be protected by the present invention. . Note that examples of cultivated vegetation will include all ornamental and flowering shrubs.

The present invention is characterized by the combined use of an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof (ie, the present culture). The mixing ratio or combination ratio of the combination is, for example, a ratio that produces a synergistic effect on the anti-nematode activity, and specifically, about 10 g to about 200 kg of amino acids, preferably about 100 g to 100 g of the main culture. A ratio of about 20 g to 4 kg can be given.
In the method of the present invention, an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof may be applied simultaneously, or an amino acid and Milotesium beru A preparation containing a culture having anti-nematode activity obtained by culturing Lucaria (Myrothecium verrucaria) or an equivalent thereof may be prepared in advance and applied.

The control agent of the present invention may be a mixture of an amino acid and a culture having anti-nematode activity obtained by culturing Myrothecium verrucaria or an equivalent thereof (that is, a main culture). Usually, however, an oil agent, emulsion, wettable powder, wettable powder, flowable agent (in water) are added by further mixing a solid carrier or liquid carrier and adding a surfactant or other auxiliary agent for formulation as necessary. Suspensions, emulsions in water, etc.), powders, granules, microcapsules and the like.
These preparations usually contain about 0.01% to 99% by weight, preferably about 5% to 95% by weight of amino acids and the main culture as the total amount of both. The mixing ratio of the amino acid and the main culture is, for example, a ratio that produces a synergistic effect on the anti-nematode activity, and specifically, about 1 to about 1 part by weight of amino acid to 10 parts by weight of the main culture. A ratio of about 20000 parts by weight, preferably about 2 parts by weight to 400 parts by weight can be given.

Examples of solid carriers used for formulation include clays (kaolin clay, diatomaceous earth, bentonite, fusami clay, acidic clay), synthetic hydrous silicon oxide, talc, ceramic, and other inorganic minerals (sericite, quartz Sulfur, activated carbon, calcium carbonate, hydrated silica, etc.), and chemical fertilizers (ammonium sulfate, phosphorous acid, ammonium nitrate, urea, ammonium chloride, etc.) and fine powders and particulates.
Examples of the liquid carrier include water, alcohols (methanol, ethanol, etc.), ketones (acetone, methyl ethyl ketone, etc.), aromatic hydrocarbons (toluene, xylene, ethylbenzene, methylnaphthalene, etc.), aliphatic hydrocarbons. (Hexane, cyclohexane, kerosene, light oil, etc.), esters (ethyl acetate, butyl acetate, etc.), nitriles (acetonitrile, isobutyronitrile, etc.), ethers (diisopropyl ether, 1,4-dioxane, etc.), acid amides (N, N-dimethylformamide, N, N-dimethylacetamide, etc.), halogenated hydrocarbons (dichloromethane, trichloroethane, carbon tetrachloride, etc.), dimethyl sulfoxide, vegetable oil (soybean oil, cottonseed oil, etc.) and the like.

  Examples of surfactants include alkyl sulfate salts, alkyl sulfonate salts, alkyl aryl sulfonate salts, alkyl aryl ethers and their polyoxyethylene compounds, polyethylene glycol ethers, polyhydric alcohol esters, and sugar alcohol derivatives. Can give.

  Other formulation adjuvants include, for example, fixing agents, dispersants, stabilizers, etc., specifically, casein, gelatin, polysaccharides (starch, gum arabic, cellulose derivatives, alginic acid, etc.), lignin derivatives, bentonite, Sugars, synthetic water-soluble polymers (polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acids, etc.), PAP (isopropyl acid phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (2- tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), mineral oils, fatty acids and fatty acid esters.

  The control agent of the present invention may be prepared by mixing a pre-formulated amino acid with a pre-formulated main culture, depending on the form of the preparation. It can also be used together.

In the present invention, for example, the effective amount or application amount of the active ingredient for controlling harmful plant parasitic nematodes is usually about 0.05 kg to about 50 kg as the effective amount or application amount of amino acids per 1000 m ^2. is there. In addition, what is necessary is just to apply as it is so that it may become said effective amount or application amount normally, when applying the active ingredient for harmful plant parasitic nematode control with a granule, a powder agent, etc. normally. Even when the active ingredient for controlling harmful plant parasitic nematodes is applied after diluting the emulsion, wettable powder, granular wettable powder, flowable powder, etc. in water, the above effective amount or application amount is obtained. So that it may be diluted with water.
In the present invention, for example, the preparation of the present invention is applied to a crop to be protected, harmful plant parasitic nematodes or habitats of harmful plant parasitic nematodes (eg, habitat or infestation of harmful plant parasitic nematodes). By applying to the soil), harmful plant parasitic nematodes can be controlled.

  The above-mentioned effective amount or application amount varies depending on the type of preparation, application time, application location, application method, pest type, damage level, etc., and should be appropriately selected by increasing / decreasing regardless of the above range. Can do.

  Hereinafter, the present invention will be described in more detail with reference examples, formulation examples and test examples, but the present invention is not limited to these examples. In addition, in a formulation example, a part represents a weight part.

Reference Example The inoculation source for the main culture is prepared by inoculating the sterilized potato-dextrose liquid medium with Milotesium velaria ATCC 46474 and culturing it at 25 ° C. for 3 to 4 weeks. 0.1 ml of the prepared inoculum was sterilized in a liquid medium (medium composition: glucose 2 g, soluble starch 15 g, yeast extract 2 g, magnesium sulfate hydrate 0.3 g, calcium carbonate 1 g, neopeptone 5 g, distilled water 1000 ml, Inoculate 100 ml of pH 7.0) and incubate with shaking at 25 ° C. for 5 days. After culturing, the obtained culture is sterilized in an autoclave (120 ° C., 1.2 atm, 20 minutes) to prepare a main culture.
Formulation Example 1
Both 15 parts of methionine and 5 parts of a preparation obtained by heat sterilization of a culture obtained by culturing Milotesium velukaria ATCC 46474 in accordance with the above reference example, 20 parts of cottonseed oil and N, N -Mix and stir in 20 parts of dimethylformamide, add 10 parts of polyethylene glycol and 10 parts of calcium dodecylbenzenesulfonate and stir well to mix and suspend to obtain an emulsion.

Formulation Example 2
Both 60 parts of methionine and 10 parts of a preparation obtained by heat sterilization of a culture obtained by culturing Myrotesium velukaria ATCC 46474 according to the above reference example, 4 parts of sodium lauryl sulfate, lignin 2 parts of calcium sulfonate, 12 parts of synthetic hydrous silicon oxide fine powder and 12 parts of diatomaceous earth are added to the mixture and mixed well to obtain a wettable powder.

Formulation Example 3
15 parts of methionine, 5 parts of a preparation obtained by heat sterilization of a culture obtained by culturing Myrotesium velukaria ATCC 46474 according to the above reference example, 5 parts of a synthetic silicon hydroxide fine powder, dodecylbenzenesulfone 5 parts of sodium acid, 30 parts of bentonite and 40 parts of clay are all well stirred and mixed, then an appropriate amount of water is added to these mixtures, stirred further, granulated with a granulator, dried by ventilation and granulated. Get.

Test example 1
It was obtained by sterilizing a culture obtained by culturing methionine (manufactured by Sumitomo Chemical Co., Ltd., hereinafter referred to as “component A agent”) and a commercially available main culture (Mirothesium versaria ATCC46474). The formulation of 90% by weight of the preparation, granulated hydrated formulation, trade name: DiTera DF, manufactured by Valent Biosciences Corporation, hereinafter referred to as “component B agent”) at a predetermined concentration with respect to 100 ml of water. Each test drug solution was prepared by dilution.
Each test chemical solution (100 ml) diluted to a predetermined concentration was applied to 5 kg of nematode-contaminated soil (nematode density: 2255 heads / 20 g (investigation method: Bellman method)) in which sweet potato root nematode (Meloidogyne incognita) was generated. Then, the soil was mixed well. 500 g of each mixed test chemical solution application soil was placed in a perforated plastic cup (860 ml, diameter: about 10 cm, height: about 11 cm), and cucumber seedlings (14 days after sowing) were transplanted (2 strains / cup, 5 repetitions) ). After transplanting, the plant was raised in a glass greenhouse.
Fifty days after transplantation, the roots of the plant that had been nurtured were washed with water, and the degree of root growth of the roots per 10 plants was calculated by observing the degree of root growth of the roots. The results are shown in Table 1. When combined use of component A agent and component B agent (invention zone), compared with the case where component A agent or component B agent is used as a single agent (comparative example zone), synergistic effect in the extent of cat root growth Was recognized. In addition, the extent of the cat root was determined as follows.

5: The plant is dead by nematode infestation.
4: A large number of bumps are found throughout the roots, forming a large chain of bumps.
3: The formation of bumps is observed in the whole root, and a large bump is formed in a part.
2: The formation of bumps is recognized in part, and some bumps are connected, but there are no large bumps. 1: The formation of bumps is slightly observed.
0: No bump formation is observed at all.

２．３

2.3

Test example 2
Each test chemical solution was prepared by diluting component A agent and component B agent to a predetermined concentration with respect to 100 ml of water.
Each test chemical solution (100 ml) diluted to a predetermined concentration was applied to 3 kg of soil contaminated with sweet potato root nematode (Meloidogyne incognita), and then the soil was mixed well. 500 g of each test chemical application soil mixed was placed in a perforated plastic cup (860 ml, diameter: about 10 cm, height: about 11 cm), and cucumber seedlings (14 days after sowing) were transplanted (2 strains / cup, 6 repetitions). ). After transplanting, the plant was raised in a glass greenhouse.
Thirty-two days after transplantation, the roots of the plant that had been nurtured were washed with water, and the control value was calculated by observing the number of goals in the root per strain. The results are shown in the table. The control value was calculated by the following formula.

Control value = (C−S) × 100 / C
C: Average number of goals per share in the untreated area S: Average number of goals per share in the treated area

INDUSTRIAL APPLICABILITY The present invention can provide an effective nematode control method and the like that can supplement or replace chemical pesticides in nematode control.

Claims (4)

A culture having an anti-nematocidal activity obtained by culturing methionine and Myrothecium verrucaria ; a sterilized product of the culture, or an extract, a partially purified product from the sterilized product, or A purified product; or an extract, a partially purified product or a purified product from the culture, or an effective amount thereof with these sterilized products , a crop to be protected, a harmful plant parasitic nematode or a harmful plant parasitic nematode A method for controlling harmful plant parasitic nematodes characterized by being applied to the habitat of the plant.   The method according to claim 1, wherein the Myrothecium verrucaria is Myrothecium verrucaria ATCC 46474 strain.   A culture having nematocidal activity obtained by culturing methionine and Myrothecium verrucaria; a sterilized product of the culture, or an extract, a partially purified product or a A harmful plant parasitic nematode control agent characterized by containing a purified product; or an extract, a partially purified product or a purified product from the culture, or a sterilized product thereof.   4. The harmful plant parasitic nematode control agent according to claim 3, wherein Myrothecium verrucaria is Myrothecium verrucaria ATCC 46474 strain.