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WO2025127025A1 - Composition de lutte contre les nématodes contenant de la l-méthionine et procédé de lutte contre les nématodes - Google Patents

Composition de lutte contre les nématodes contenant de la l-méthionine et procédé de lutte contre les nématodes Download PDF

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WO2025127025A1
WO2025127025A1 PCT/JP2024/043595 JP2024043595W WO2025127025A1 WO 2025127025 A1 WO2025127025 A1 WO 2025127025A1 JP 2024043595 W JP2024043595 W JP 2024043595W WO 2025127025 A1 WO2025127025 A1 WO 2025127025A1
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methionine
nematode
nematodes
soil
ppm
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Japanese (ja)
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隆之 水久保
泰裕 鎌田
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Maruwa Biochemical Co Ltd
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Maruwa Biochemical Co Ltd
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/12—Powders or granules
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P5/00—Nematocides

Definitions

  • Plant parasitic nematodes are nematodes that parasitize plants and are known to cause damage such as inhibiting plant growth and causing plant death.
  • plant parasitic soil nematodes such as root-knot nematodes, root-lesion nematodes, and cyst nematodes are one of the factors that inhibit agricultural crop production, and nematode damage accounts for 16% of the causes of continuous crop damage in Japan. Furthermore, the average yield reduction due to nematodes in all crops around the world is 12%. Meanwhile, damage caused by plant parasitic nematodes is also known to occur in the production and use of useful plants such as lawns and shrubs. Therefore, it is an urgent task to provide an effective method for controlling nematodes in order to reduce damage caused by nematodes to plants and to increase crop productivity.
  • Non-Patent Document 1 Non-Patent Document 1
  • D-D agents are restricted by the EPA in the United States due to their carcinogenicity and other issues, and their pesticide registration in the EU has already expired.
  • Chloropicrin is also highly irritating to the human body, so its use in crowded environments tends to be avoided.
  • soil reduction disinfection one of the agricultural methods, is expected to contribute to the promotion of sustainable agriculture because of its low environmental impact, but there are many restrictions on its implementation and it has not been widely adopted.
  • Non-Patent Document 2 the nematode Acanthurus nigricans
  • Non-Patent Document 6 the wheat cyst nematode
  • Non-Patent Document 6 the peel nematode
  • Non-Patent Document 7 Patent Documents 2, and 3
  • a species of the genus Veronoraimus Non-Patent Document 8
  • a species of the genus Mesocrichonema Non-Patent Document 8
  • Non-Patent Document 7 reports that the toxicity of D- and L-methionine to potato cyst nematodes was equivalent, and the mechanism of action is said to be oral toxicity.
  • Patent Document 2 reports that the oral toxicity of D-methionine to potato cyst nematodes was lower than that of L-methionine.
  • Non-Patent Document 3 cites Non-Patent Document 4 and states that while the DL-methionine showed contact toxicity to sweet potato root-knot nematodes, the L-methionine did not show contact toxicity.
  • JP 2000-007506 A International Publication No. 2019/004252 JP 2022-118806 A
  • Nematologica 13 318-323. Evans, K. and Trudgill, D.L. 1971 Effects of amino acids on the reproduction of Heterodera rostochiensis. Nematologica 17: 495-500. Crow, W.T., Cuda, J.P., Stevens, B.R. 2009. Efficacy of Methionine Against Ectoparasitic Nematodes on Golf Course Turf. Journal of Nematology, 41: 217-220. Epstein, E. 1973.
  • the objective of this disclosure is to provide a nematode control composition and control method that exhibits high control activity even when the application rate of methionine is small.
  • the inventors discovered that when L-methionine is applied, it is effective in controlling nematodes even at low concentrations, and thus completed the disclosed nematode control composition and control method.
  • a first aspect of the present disclosure is a method for manufacturing a semiconductor device comprising: A composition for controlling nematodes comprising methionine, The methionine comprises more than 50% by weight of L-methionine based on the total weight of the methionine.
  • a second aspect of the present disclosure is a method for manufacturing a semiconductor device comprising: A method for controlling nematodes, comprising the steps of: applying methionine in an amount of 5 to 80 kg per 1000 m2 of field; The methionine comprises more than 50% by weight of L-methionine based on the total weight of the methionine.
  • the nematode control composition disclosed herein contains methionine, and since the methionine contains L-methionine, it can control nematodes safely, inexpensively, and effectively. Furthermore, the nematode control composition disclosed herein has a small environmental impact and can be applied easily, so it can contribute to the promotion of sustainable agriculture.
  • the nematode control method of the present disclosure applies 5 to 80 kg of methionine per 10 a (1000 m2 ) of field, and the methionine contains L-methionine, thereby enabling nematodes to be controlled safely, inexpensively, and effectively compared to conventional control methods. Furthermore, the method of the present disclosure has the advantage that, since the amount of methionine applied is smaller than that of conventional methods, it is not toxic to plants and can be carried out during the cultivation period. Furthermore, the method of the present disclosure has a small environmental impact and is simple, and can therefore contribute to the spread of sustainable agriculture.
  • the nematode-inhibiting effect of an aqueous methionine solution was evaluated using the method described in Example 1, and the results are shown in terms of the relationship between methionine concentration and survival rate.
  • (A) shows the results after 1 day
  • (B) shows the results after 4 days.
  • the results are shown as time-dependent changes in the nematode-inhibiting effect of an aqueous methionine solution evaluated by the method described in Example 1.
  • (A) shows the results at 2500 ppm
  • (B) shows the results at 250 ppm. 1 shows the results of evaluating the effect of the ratio of L-methionine to D-methionine on the inhibitory effect against Meloidogyne incognita, using the method described in Example 2.
  • (A) shows the results at 300 ppm, (B) at 600 ppm, and (C) at 1200 ppm.
  • the nematode-inhibiting effect of an aqueous methionine solution was evaluated by the method described in Example 4, and the results are shown in terms of the relationship between the methionine concentration and the survival rate.
  • 1 shows the results of evaluating the inhibitory effect of an aqueous methionine solution against Meloidogyne root-knot nematode by the method described in Example 5.
  • (A) shows the results after 1 day, and (B) shows the results after 4 days.
  • the results of evaluating the inhibitory effect of an aqueous methionine solution against Meloidogyne arenaria knot nematode are shown in terms of the relationship between methionine concentration and survival rate by the method described in Example 6.
  • (A) shows the results after 3 hours
  • (B) shows the results after 2 days
  • (C) shows the results after 3 days.
  • the control effects of optical isomers of methionine on Meloidogyne incognita were compared using the method described in Example 7, and the results are shown below. Photographs of tomato roots harvested from the test described in Example 7.
  • (A) are two control tomato roots
  • (B) are two tomato roots spiked with 67 ppm D-methionine
  • (C) are, from the right, roots from tomato roots spiked with 67 ppm, 133 ppm, and 200 ppm D-methionine.
  • Photographs of tomato roots harvested from the test described in Example 7. (A) shows, from the right, tomato roots supplemented with 67 ppm, 133 ppm, and 200 ppm L-methionine.
  • (B) shows, from the right, tomato roots supplemented with 67 ppm, 133 ppm, and 200 ppm DL-methionine.
  • compositions for controlling nematodes comprising methionine.
  • the composition of the present disclosure may comprise 20 to 100% by weight, preferably 40 to 100% by weight, more preferably 60 to 100% by weight of methionine.
  • the composition of the present disclosure may consist of methionine alone.
  • the methionine comprises L-methionine.
  • the composition of the present disclosure comprises 50 to 100% by weight, preferably 75 to 100% by weight, more preferably 95 to 100% by weight of L-methionine, based on the total weight of the composition.
  • Methionine is one of the 20 amino acids that make up proteins, and is an essential amino acid. It exists in D-form (D-methionine) and L-form (L-methionine), and when the D-form and L-form exist in equal amounts, it is called a racemic mixture (DL-methionine, DL-form). Methionine is widely used as a human medicine, veterinary medicine, food additive, and feed additive, and is a highly safe substance. Furthermore, it is less expensive than existing pesticides.
  • the methionine comprises L-methionine.
  • the methionine comprises more than 50% by weight, preferably 75% by weight or more, more preferably 85% by weight or more, even more preferably 95% by weight or more, and most preferably 99% by weight or more of L-methionine based on the total weight of methionine.
  • the methionine comprises up to 100% by weight of methionine.
  • the methionine may consist only of L-methionine.
  • the L-methionine in this disclosure may be produced by any method.
  • methods for producing L-methionine include a synthetic method, an enzymatic method, an extraction method, and a fermentation method.
  • the synthetic method is a method for isolating only L-methionine from a racemic form of methionine that has been chemically synthesized by a known method.
  • the enzymatic method is a method for converting a precursor to L-methionine by an enzyme reaction.
  • the extraction method is a method for isolating only L-methionine from a mixture of amino acids obtained by decomposing proteins.
  • the fermentation method is a method for selectively producing only L-methionine by fermenting the raw material using a microorganism.
  • the methionine does not contain D-methionine. If present, it is desirable that the amount of D-methionine is less than 50% by weight, preferably 25% by weight or less, more preferably 15% by weight or less, even more preferably 5% by weight or less, and most preferably 1% by weight or less, based on the total weight of methionine. In one embodiment, the compositions of the present disclosure do not contain D-methionine.
  • the composition of the present disclosure may further contain a surfactant.
  • a surfactant By containing a surfactant in the composition of the present disclosure, it is possible to promote the penetration of methionine into voids in the soil and promote the contact between L-methionine and the target nematode.
  • the composition of the present disclosure may contain the surfactant in an amount of 0.1 to 20% by weight, preferably 0.1 to 10% by weight.
  • Surfactants that can be used in the composition of the present disclosure include, for example, nonionic surfactants, anionic surfactants, cationic surfactants, etc.
  • Nonionic surfactants include, but are not limited to, polyoxyethylene alkyl ethers, polyoxyethylene alkyl phenyl ethers, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, sucrose fatty acid esters, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, alkyl glycosides, etc.
  • Anionic surfactants include alkyl sulfates, alkyl ether sulfates, lignin sulfonates, etc.
  • Cationic surfactants include, for example, quaternary alkyl ammonium salts, etc.
  • composition of the present disclosure may further contain additional components, such as a bulking agent, a solvent, a pH adjusting agent, a binder, a disintegrant, a physical property improving agent, an antifungal agent, a stabilizer, a colorant, a fragrance, a chemical harm reducing agent, and the like.
  • additional components such as a bulking agent, a solvent, a pH adjusting agent, a binder, a disintegrant, a physical property improving agent, an antifungal agent, a stabilizer, a colorant, a fragrance, a chemical harm reducing agent, and the like.
  • bulking agents include, but are not limited to, talc, bentonite, diatomaceous earth, amorphous silica, clay, attapulgite, calcium carbonate, and other inorganic salts.
  • Solvents include, but are not limited to, water, methanol, and ethanol.
  • pH adjusters include, but are not limited to, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, lactic acid, sodium lactate, phosphoric acid, sodium dihydrogen phosphate, citric acid, and sodium citrate.
  • Binders include, but are not limited to, polyvinylpyrrolidone, polyvinyl alcohol, carboxymethylcellulose, sodium carboxymethylcellulose, starch, starch derivatives, sodium acrylates, oils and fats (e.g., vegetable oils such as soybean oil, palm oil, and corn oil, and animal oils such as lard, fish oil, and butter).
  • oils and fats e.g., vegetable oils such as soybean oil, palm oil, and corn oil, and animal oils such as lard, fish oil, and butter).
  • the composition of the present disclosure can be in any form.
  • the composition of the present disclosure can be in the form of a solid or liquid that can be directly mixed or sprayed on soil, or in the form of a solid or liquid that can be dissolved in a solvent such as water before use.
  • the form of the composition of the present disclosure includes, but is not limited to, dust, granule, powder, wettable powder, water-soluble, emulsion, liquid (including solution), oil, aerosol, coating agent, microcapsule, etc.
  • the composition of the present disclosure is preferably in the form of a dust, granule, powder, wettable powder, or water-soluble, and more preferably in the form of a granule, powder, powder, or water-soluble.
  • the composition of the present disclosure can be prepared by any method known in the art.
  • the composition of the present disclosure can be prepared by using a process such as grinding, mixing, classification, granulation, etc., alone or in combination.
  • the composition of the present disclosure can be obtained in the form of a solution by, for example, a dissolving process, or the composition of the present disclosure can be obtained in the form of a suspension or emulsion by, for example, a dispersing process.
  • the raw powder of methionine can be used as the composition of the present disclosure.
  • the milling process is a process in which bulk methionine powder is milled to a desired particle size by methods known in the art.
  • the milled methionine may be used as a composition of the present disclosure or may be provided to further processes.
  • the blending step is a step in which raw or milled methionine is blended with additional ingredients, such as bulking agents, by methods known in the art.
  • additional ingredients such as bulking agents
  • the blend of methionine and additional ingredients may be a composition of the present disclosure, or the blend may be subjected to further steps.
  • the classification process is a process in which methionine or a raw material containing methionine is separated according to the particle size of the powder or particles.
  • Classification methods are broadly classified into dry classification and wet classification, both of which are known in the art.
  • the methionine or a composition containing methionine obtained by the classification process may be used as the composition of the present disclosure, or may be further provided to another process.
  • the granulation process is a process in which methionine or a mixture of methionine and additional ingredients is processed to a desired particle size.
  • Granulation methods are broadly classified into wet granulation and dry granulation, and are known in the art.
  • the resulting granulated product may be used as the composition of the present disclosure, or may be further subjected to another process.
  • the dispersion process is a process in which methionine or a raw material containing methionine in a solid or liquid form is dispersed in a solvent or oil.
  • the methionine or a raw material containing methionine may be obtained through the above-mentioned processes of grinding, mixing, classification, granulation, etc.
  • the dispersion process can be carried out by any method known in the art.
  • the obtained dispersion may be used as the composition of the present disclosure, or may be further provided to another process.
  • Nematodes are a general term for animals belonging to the phylum Nematoda, and are long, slender worms with body lengths of 0.1 mm to 1000 mm. Nematodes live in large numbers in all biospheres on Earth, and are divided into free-living and parasitic species. In the agricultural field, a group of parasitic species called plant-parasitic nematodes (also called harmful nematodes or plant-pathogenic nematodes) are problematic because they cause agricultural damage.
  • the composition of the present disclosure can be suitably used for controlling plant parasitic nematodes, more specifically, plant parasitic nematodes that live in soil.
  • Nematodes that can be controlled by the composition of the present disclosure include, but are not limited to, root-knot nematodes, root-leach nematodes, and cyst nematodes. Examples of root-knot nematodes include Meloidogyne incognita, Meloidogyne nervosa, Meloidogyne arenaria, and Meloidogyne javanica.
  • Root-leach nematodes examples include Meloidogyne nervosa, Meloidogyne nervosa, Meloidogyne nervosa, Meloidogyne nervosa, and Meloidogyne nervosa.
  • cyst nematodes examples include the potato cyst nematode, the potato white cyst nematode, the soybean cyst nematode, the clover cyst nematode, and the sugar beet cyst nematode.
  • Other examples include the nematode Nematode gracilis, Nematode umbellata, and Nematode nigricans.
  • the composition of the present disclosure can be applied to soil by any method known in the art.
  • the composition of the present disclosure may be applied before planting the plant or after planting. It may also be applied both before and after planting the plant.
  • the composition of the present disclosure is applied after planting, it is preferable to apply it 20 to 60 days, preferably 25 to 50 days, more preferably 30 to 40 days, and most preferably 30 days after planting. It is preferable that the soil to which the composition of the present disclosure is applied is uniformly mixed.
  • the term "plant” includes plants cultivated in fields, i.e., crops, and useful plants other than crops, such as lawns and trees.
  • the term “useful plant” refers to a plant that is useful for human life.
  • composition of the present disclosure can be applied in an amount corresponding to 5 to 40 kg, preferably 7.5 to 30 kg, and more preferably 10 to 20 kg of L-methionine per 10 a (1000 m2 ) of field. This range is preferable because it does not inhibit plant growth and can effectively control nematodes.
  • field includes not only a place where crops are cultivated, but also a place where useful plants other than crops are grown.
  • composition of the present disclosure may be mixed with a surfactant at the time of application, or may be dissolved or dispersed in a solvent or oil.
  • the composition of the present disclosure may be in a solid or liquid form.
  • the surfactant, solvent, and oil are as described in the (Additional Components) section.
  • composition disclosed herein contains L-methionine as an active ingredient, and is therefore highly safe, inexpensive, and effective in controlling nematodes.
  • the composition disclosed herein requires a smaller amount of methionine than conventional compositions, and therefore has the advantage that it does not inhibit plant growth and can be applied during the cultivation period.
  • the composition disclosed herein has a small environmental impact and can be applied easily, making it possible to contribute to the spread of sustainable agriculture.
  • the second aspect of the present disclosure relates to a method for controlling nematodes.
  • the method of the present disclosure includes applying methionine in an amount of 5 to 80 kg, preferably 7.5 to 60 kg, more preferably 10 to 40 kg per 10 a (1000 m 2 ) of field, the methionine comprising more than 50% by weight of L-methionine based on the total weight of methionine.
  • the methionine comprises up to 100% by weight of L-methionine based on the total weight of methionine.
  • Methionine is as described in the section (methionine) of "1. Composition".
  • methionine can be applied by any method known in the art.
  • methionine may be applied to soil in a powdered or granular solid form, or methionine may be dissolved in a solvent and then applied.
  • Methionine or a solution thereof may be mixed with a surfactant at the time of application.
  • methionine formulated by a method known in the art may be applied. Methods for formulating methionine are described in the section "1. Composition" (Methionine Manufacturing Method), and surfactants are described in the section “Additional Components.”
  • methionine may be applied before or after planting of the plant.
  • the methods of the present disclosure include applying methionine within 20 days, preferably within 15 days, and more preferably within 10 days before planting.
  • the methods of the present disclosure include applying methionine 20-60 days, preferably 25-50 days, more preferably 30-40 days, and most preferably 30 days after planting.
  • the methods of the present disclosure include applying methionine before planting and applying methionine after planting.
  • the method of the present disclosure preferably includes uniformly mixing the soil after application of methionine.
  • the depth of the soil to be mixed can be 20 cm, more preferably 15 cm, from the soil surface.
  • the method of the present disclosure may further include applying a surfactant.
  • Surfactants that can be used in the method of the present disclosure are as described in the section "1. Composition" (Surfactant).
  • the surfactant may be applied before or after the methionine. Also, by blending methionine and a surfactant, they can be applied simultaneously.
  • the method disclosed herein is highly safe, inexpensive, and effective in controlling nematodes. Furthermore, the method disclosed herein has the advantage that it requires a smaller amount of methionine to be applied than conventional methods, is not toxic to plants, and can be carried out during the cultivation period. Furthermore, the method disclosed herein has a small environmental impact and is simple, so it can contribute to the spread of sustainable agriculture.
  • Example 1 Comparison of the nematode suppression effect of methionine aqueous solution by optical isomers (1) Aqueous solutions of D-, L-, and DL-methionine at various concentrations were contacted with Meloidogyne incognita to evaluate the inhibitory effect of each optical isomer against the nematode.
  • Test method 1 Meloidogyne incognita (MAFF108258 strain) was added to tap water to prepare a nematode suspension at 250 heads/mL, which was then stored at 10° C. until use. Note that Meloidogyne incognita hatched larvae were collected 6 days after egg capsule collection and immediately used to prepare the suspension.
  • D-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%), L-methionine (Asuka Animal Health Co., Ltd., purity 99.6%), and DL-methionine (Asuka Animal Health Co., Ltd., purity 99.5%) were each dissolved in purified water to prepare aqueous methionine solutions with concentrations of 0.5 ppm, 5 ppm, 50 ppm, 500 ppm, and 5,000 ppm.
  • 100 ⁇ L of nematode suspension was poured into each well of a 96-well plate (round bottom, water-repellent coating) using a glass pipette.
  • Tables 1-2 and Figures 1-2 show the relationship between the concentration of each isomer of methionine and the survival rate of nematodes after 1 day and 4 days.
  • Table 2 and Figure 2 show the change in survival rate of nematodes over time for 2500 ppm and 250 ppm. Each value is an average value.
  • the results show that the L-methionine form showed concentration-dependent contact toxicity to Meloidogyne incognita one day after immersion, whereas the D- and DL-methionine forms did not. However, after four days of immersion, the D- and DL-methionine forms also showed concentration-dependent contact toxicity. Looking at the results by concentration, at a concentration of 250 ppm, the time course of survival rates of nematodes exposed to the L- and DL-methionine forms showed a similar tendency, reaching around 40% after four days, but not reaching 0%.
  • the survival rate of the L-methionine form reached 0% after four days, whereas the D- and DL-methionine forms were around 60% after four days and did not reach 0% even after seven days.
  • the LD 50 of the DL form should be 120 ppm, which is twice that of the L form, but in reality it was about 9 times that value. From this, it was suggested that, without being bound by theory, the D-isomer may inhibit the inhibitory effect of the L-isomer against nematodes.
  • the aqueous solution concentration (ppm) was converted to the amount of soil to be treated in the field as follows.
  • the rotary tillage depth is about 15 cm, so if the depth of the field to be mixed is 15 cm, the amount of soil per 10 a (1000 m2 ) is 150,000 kg. Since soil also contains a lot of air, if its density is assumed to be 1 kg/L, the volume is 150,000 L.
  • Example 2 Effect of the Mixture Ratio of L- and D-Methionine on Inhibition of Meloidogyne incognita Since the results of Example 1 suggested that the D-methionine may inhibit the inhibitory effect of the L-methionine on nematodes, the mixture ratio of the L- and D-methionine was gradually changed to evaluate the effect of the D-methionine on the nematode inhibitory effect of the L-methionine.
  • Test method (1) Meloidogyne incognita (strain MAFF108258) was added to tap water to prepare a nematode suspension at 250 nematodes/mL, which was stored at 10°C until use. (2) A total of 0.24 g of L-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%) and D-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%) were dissolved in 100 mL of purified water to give ratios (weight ratios) of 100:0, 99:1, 95:5, 85:15, 75:25, and 50:50, respectively, to prepare 6 levels of 2400 ppm methionine aqueous solutions with a ratio of L-form to the total methionine ranging from 100% by weight to 50% by weight.
  • a 1200 ppm aqueous solution was prepared by diluting 10 mL of this 2400 ppm aqueous solution with 10 mL of purified water. Furthermore, a 600 ppm aqueous solution of methionine was prepared by diluting 5 mL of the 2400 ppm aqueous solution with 15 mL of purified water. (3) 100 ⁇ L of the nematode suspension was poured into each well of a 96-well plate (round bottom, water-repellent coating) using a glass pipette. In addition, 100 ⁇ L of methionine solution of each concentration was added to each well.
  • the methionine concentrations when the nematodes were immersed were 300 ppm, 600 ppm, and 1200 ppm, respectively.
  • 100 ⁇ L of purified water was added to the control (0 ppm, CTL).
  • the number of surviving nematodes was counted under transmitted illumination using a stereomicroscope immediately after immersion in the methionine aqueous solution (0 days), and 1, 3, and 6 days after immersion. The number of surviving nematodes was determined in the same manner as in 1. Test method (4) of Example 1.
  • the aqueous solutions with an L-isomer proportion of 99% by weight and 100% by weight exhibited a high nematode suppression effect at all concentrations, and at 600 ppm (corresponding to a field treatment amount of 30 kg/10a) and 1200 ppm (corresponding to a field treatment amount of 60 kg/10a), the nematode survival rate after 6 days was almost 0%. From these results, it was shown that the composition of the present disclosure can provide an excellent nematode suppression effect by containing more than 50% by weight of L-methionine based on the total weight of methionine, and that the effect is enhanced as the proportion of L-isomers increases.
  • Example 3 Comparison of nematode suppression effects of methionine aqueous solution using optical isomers (2) From the results of Example 1, it was found that in the case of L-methionine, the survival rate of nematodes changed significantly between 250 ppm and 2500 ppm. Therefore, the concentration of the methionine solution was changed and tests were conducted in the same manner as in Example 1 to evaluate in more detail the relationship between the methionine concentration and the nematode suppression effect.
  • Test Method The test was carried out in the same manner as in Example 1, except that aqueous methionine solutions were prepared at concentrations of 600 ppm, 1200 ppm, 1800 ppm, 2400 ppm, and 5000 ppm.
  • Example 4 Comparison of nematode suppression effects of methionine aqueous solution using optical isomers (3)
  • Examples 1 to 3 showed the inhibitory effect of L-methionine when it was in contact with nematodes for a long period of time. Therefore, tests were conducted with a shorter contact time to evaluate the inhibitory effect of nematodes (Meloidogyne incognita) when it was in contact for a short period of time.
  • Test Method The test was carried out in the same manner as in Example 1, except that the concentrations of the methionine aqueous solution were 0 ppm, 300 ppm, 600 ppm, 1200 ppm, 1800 ppm, 2400 ppm, and 4800 ppm, and the test time was 4 hours. The test was carried out in duplicate.
  • Example 5 Evaluation of Inhibitory Effect of Methionine on Meloidogyne northern root-knot nematode
  • Meloidogyne northern root-knot nematode was used to evaluate the inhibitory effect of each optical isomer of methionine.
  • Test method The test was conducted in the same manner as in Example 1, except that the concentrations of the prepared methionine aqueous solutions were 300 ppm, 600 ppm, 1200 ppm, and 1800 ppm, and Meloidogyne nigra (MAFF108260 strain) was used as the nematode. The number of surviving nematodes was counted 1 day and 4 days after the start of the test.
  • L-methionine After one day, only L-methionine showed an inhibitory effect against Meloidogyne. After four days, L-methionine showed an excellent inhibitory effect against Meloidogyne. On the other hand, D-methionine and DL-methionine showed no inhibitory effect after one day, and after four days, a decrease in survival rate was observed, but the effect was slight.
  • Example 6 Evaluation of Inhibitory Effect of Methionine on Meloidogyne arenaria Root-knot Meloidogyne The control effect of each optical isomer of methionine was evaluated using Meloidogyne arenaria as the nematode.
  • Test method As the optical isomers of methionine, D-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%), L-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%) and DL-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%) were used.
  • the test was conducted in the same manner as in Example 1, except that the concentrations of the prepared methionine aqueous solutions were 300 ppm, 600 ppm, 1200 ppm, 1800 ppm, 2400 ppm and 4800 ppm, and that Meloidogyne arenaria (MAFF108262 strain) was used as the nematode. The number of surviving nematodes was counted 3 hours, 2 days and 3 days after the start of the test.
  • the D- and DL-methionine forms showed high survival rates of nematodes at all concentrations.
  • the L-methionine form showed a high nematode suppression effect, and the effect increased roughly in proportion to the concentration.
  • the survival rate of nematodes treated with the D-methionine form was maintained at a high level.
  • the survival rate of nematodes treated with the DL-methionine form decreased in a concentration-dependent manner up to 1200 ppm, but at 2400 ppm, the survival rate recovered and showed a high value.
  • the presence of a high concentration of the D-methionine form nematode suppressed the suppressive effect of the L-methionine form against the root-knot nematode.
  • the survival rate of nematodes treated with the L-methionine form was maintained at a low level at all concentrations. This result indicates that the composition of the present disclosure also shows a high suppressive effect against the root-knot nematode.
  • Example 7 Comparison of the control effects of optical isomers of methionine against Meloidogyne incognita D-, L- and DL-methionine were added at various concentrations to soil inoculated with Meloidogyne incognita, and the effect on the degree of damage caused by root clubs in tomatoes was evaluated .
  • Test method Soil in which Meloidogyne incognita (strain MAFF108258, purchased from the National Agriculture and Food Research Organization Gene Bank) was grown, commercially available black soil, and horticultural nursery soil (manufactured by Hokusan Co., Ltd.) were mixed in a ratio of 1:1:1 to prepare nematode-contaminated soil.
  • D-methionine (Tokyo Chemical Industry Co., Ltd., purity >99.0%), L-methionine (Asuka Animal Health Co., Ltd., purity 99.6%), and DL-methionine (Asuka Animal Health Co., Ltd., purity 99.5%) were each dissolved in ion-exchanged water to a concentration of 0.5% (w/v) to prepare a 0.5% aqueous methionine solution.
  • 3 8 mL, 16 mL, and 24 mL of 0.5% methionine solution were added to 600 mL of nematode-infested soil and mixed.
  • Test method (1) Meloidogyne arenaria (MAFF108262 line, obtained from the National Agriculture and Food Research Organization Gene Bank) was propagated on tomatoes planted in black soil. The soil was sieved through a 2.5 mm mesh screen to remove the roots. (2) A portion of the soil prepared in step (1) was removed, and the number of nematodes in the soil was measured by isolating the nematodes from the soil using the Bellman funnel method. First, 20 g of soil was placed on a mesh plate covered with paper, immersed in a glass funnel filled with water, and left to stand for three days to separate the nematodes into a vial attached to the leg of the glass funnel.
  • MAFF108262 line obtained from the National Agriculture and Food Research Organization Gene Bank
  • nematode larvae that had settled in the vial were collected, and the number of individuals was counted using a plankton counting slide.
  • An appropriate amount of the sieved soil from step (1) was mixed with commercially available black soil whose moisture content had been adjusted to 33% to prepare nematode-infested soil with an initial density of 2.5 nematodes per gram of soil.
  • Control value (%) (1 - average number of nematodes under each condition / average number of nematodes in the control) x 100
  • L-methionine showed a higher control value than D-methionine.
  • the nematode density was 0 at a treatment concentration of 30 kg/10a of L-methionine.
  • the control value was lower with D-methionine, and at a treatment concentration of 15 kg/10a the control value was negative.
  • Example 9 Comparison of the effects of optical isomers of methionine on suppressing root-knot nematode damage in sweet potato Since the demand for nematode control agents is particularly high in sweet potato cultivation, L- and DL-methionine were added to soil inoculated with Meloidogyne incognita, and the effect on the degree of root-knot damage in sweet potato was evaluated.
  • Test method (1) Meloidogyne incognita (MAFF108258 line, obtained from the National Agriculture and Food Research Organization Gene Bank) was propagated on tomatoes planted in black soil. The soil was sieved through a 2.5 mm mesh screen to remove the roots. (2) Nematodes were isolated using the same procedure as the Bellman funnel method described in step (2) of Example 8, and the number of individuals was counted using a plankton counting slide. (3) Test nematode-infested soil was prepared in the same manner as in (3) of Example 8. The initial density of nematodes was 1.25 nematodes per gram of soil.
  • the nematode-infested soil containing the L- and DL-nematodes was filled into a 3-L planter, and a single sweet potato vine (variety: Takakei 14) was planted in each.
  • the control was an untreated control (CTL), in which soil not containing methionine was used.
  • CTL untreated control
  • the planters were placed in a glass chamber and grown for 52 days (maximum temperature 35°C, average temperature 28°C). During the growth period, the soil was watered once a day to prevent the soil surface from drying out. (7) 52 days after the start of cultivation, the roots were removed from the pots.
  • the soil was removed by rinsing in the shower, and the vines and potatoes were removed from the roots.
  • the washed roots were stored in a freezer at -20°C until the number of root galls was counted.
  • the thawed roots were cut into 5 mm lengths, suspended in water, transferred to a tea strainer, and squeezed to remove the water.
  • the weight (g) of the entire roots was measured, and 2 g was taken from it to count the number of root galls and egg sacs.
  • the egg sacs included those whose presence was estimated from the peeling traces left on the root galls.
  • the number of root galls was equal to the number of egg sacs when two or more egg sacs or their peeling traces were found on one lump.
  • the number of root galls and egg sacs per plant was calculated by multiplying the number of root galls and egg sacs per 2 g by the weight (g) of the entire roots, and then dividing by 2.
  • Control value (%) (1 - average number of galls or egg sacs / average number of galls or egg sacs in the control) x 100
  • Example 10 Comparison of nematode-inhibiting effects of L-methionine powder and aqueous solution L-methionine was applied to soil in different forms to evaluate its effect on nematode-inhibiting effects.
  • Nematode Meloidogyne incognita (strain MAFF108258) was mixed with commercially available sterilized andosol to give a nematode density of 20 nematodes/20 g soil, to prepare nematode-contaminated soil.
  • Nematode-infested soil was treated with L-methionine under the following conditions A to C.
  • the control (CTL) was soil that was not treated with L-methionine.
  • A Powder mixing treatment 4 g of sand in which L-methionine had been diluted 100-fold was mixed with 600 mL of nematode-infested soil so that the amount of L-methionine was 67 ppm (corresponding to a field treatment amount of 10 kg/10 a). Water was added to adjust the soil moisture content to 32%, and the mixture was placed in a plastic bag and allowed to stand at 25°C for 3 days in a sealed state. Four days after mixing with L-methionine, the soil was filled into a pot and used to grow tomatoes.
  • B Aqueous solution mixing treatment
  • L-methionine was dissolved in ion-exchanged water to prepare a 0.5% methionine aqueous solution.
  • the results are shown in Table 17.
  • the number of root clubs is the average value, and the ratio to CTL indicates the ratio (percentage) of the number of root clubs under each condition to the number of root clubs in the control.
  • the control value is the value obtained by subtracting the ratio to CTL from 100%.
  • L-methionine showed superior inhibitory effects compared to the control, but especially under conditions A (powder mixing treatment) and B (aqueous solution mixing treatment), it showed a high inhibitory effect.
  • Example 11 Evaluation of nematode suppression effect by L-methionine The effect of suppressing root clubs was evaluated by changing the amount of L-methionine applied to the soil in powder form. A similar test was also performed with DL-methionine to compare the suppression effects.
  • Test method (1) Meloidogyne incognita (MAFF108258 line) was grown in cherry tomatoes (variety: Regina) in a planter filled with andosol. The soil in which Meloidogyne incognita had grown was diluted with commercially available sterilized andosol to prepare nematode-contaminated soil with a nematode density of 20 nematodes/20 g soil. (2) L-methionine powder was diluted 100-fold with sand, and 3 g and 4 g of sand were mixed with 600 mL of nematode-contaminated soil.
  • the L-methionine concentrations were 50 ppm and 67 ppm (equivalent to 7.5 kg and 10 kg per 10 a of soil, respectively, indicated as “L7.5” and “L10", respectively).
  • DL-methionine powder was mixed with 600 mL of soil to a concentration of 67 ppm (DL10).
  • the control was soil without methionine (CTL).
  • CTL methionine
  • the soil was placed in a plastic bag and left to stand for 3 days in a sealed state.
  • CTL methionine
  • Each soil was filled into the pot.
  • One cherry tomato (variety: Regina) was planted in each pot 31 days after sowing, and grown for 24 days under LED light for plant growth in a thermostatic room at 25°C. After 24 days, each root was removed from the pot, the soil was washed off with a shower, and the number of root galls was counted under a stereomicroscope.

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Abstract

La présente divulgation consiste à fournir : une composition de lutte contre les nématodes qui présente une activité de régulation élevée même lorsque la quantité de méthionine appliquée est faible; et un procédé de contrôle. La présente divulgation concerne une composition de lutte contre les nématodes qui contient de la méthionine, dans laquelle la méthionine comprend plus de 50% en poids de L-méthionine par rapport au poids total de la méthionine. De plus, la présente invention concerne également un procédé de lutte contre les nématodes comprenant l'application de 5 à 80 kg de méthionine pour 1 000 m2 d'un champ.
PCT/JP2024/043595 2023-12-11 2024-12-10 Composition de lutte contre les nématodes contenant de la l-méthionine et procédé de lutte contre les nématodes Pending WO2025127025A1 (fr)

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WO2019004252A1 (fr) * 2017-06-28 2019-01-03 株式会社シティック Procédé de régulation de globodera rostochiensis

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JP2000007506A (ja) * 1998-06-16 2000-01-11 Hodogaya Chem Co Ltd 土壌線虫相の改善方法
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WO2019004252A1 (fr) * 2017-06-28 2019-01-03 株式会社シティック Procédé de régulation de globodera rostochiensis

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Title
OSMAN A. A. , VIGLIERCHIO DAVIDE R.: "Meloidogyne incognita development on soybean treated with selected amino acids by alternate methods", REVUE DE NEMATOLOGIE, vol. 4, no. 1, 1 January 1981 (1981-01-01), pages 172 - 174, XP093324192 *

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