Substance for promoting incubation of cyst nematodes (incubation promoting substance)

Abstract

The present disclosure provides a substance (incubation promoting substance) that promotes incubation of cyst nematodes. According to the present invention, the incubation promoting substance death cyst nematodes by hunger after incubation by incubating the cyst nematodes in the soil without the presence of parasitic plants, and thus can be advantageously used to prevent damage or the like of the cyst nematodes to crops.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to a substance that promotes hatching of cyst nematodes (hatching promotion substance). The present disclosure also relates to a method of examining a substance that promotes hatching of cyst nematodes using eggs of the cyst nematodes. BACKGROUND

[0002] Cyst nematodes can survive in soil for several to 10 years, hatch depending on the presence of a plant on which they can parasitize, parasitize the plant, and cause damage to the plant. Female nematodes die in a state in which several hundred eggs are contained in an egg case, and the carcasses form cysts. Cysts exhibit resistance to drugs, temperature, dryness, and the like, and are capable of protecting the eggs from the external environment. Cysts also have resistance to pesticides, and the effect of crop rotation is low, and it is not easy to eradicate them.

[0003] It is considered that if hatching of cyst nematodes derived from cysts is promoted in an environment in which a plant on which they can parasitize is not present, the cyst nematodes cannot take in nutrients based on parasitism after hatching, and will starve to death. Therefore, for the purpose of eradicating cyst nematodes, a hatching promotion substance for nematodes is being developed. Potato cyst nematodes hatch by Solanoeclepin A, which is a hatching promotion substance secreted from potatoes. However, the total synthesis of Solanoeclepin A is difficult (Non-Patent Literature 1). In Patent Literature 1, as a hatching attractant for nematodes, plants infected with cyst nematodes, respectively, are disclosed.

[0004] PRIOR ART DOCUMENTS

[0005] PATENT LITERATURE

[0006] Patent Literature 1: JP 2021-036984 A

[0007] NON-PATENT LITERATURE

[0008] Non-Patent Literature 1: Tanino K. et. al., Nature Chemistry, 3, 484-488, 2011 SUMMARY

[0009] The present disclosure provides a substance that promotes hatching of cyst nematodes (hatching promotion substance). The present disclosure also provides a method of examining a substance that promotes hatching of cyst nematodes using eggs of the cyst nematodes.

[0010] In the present disclosure, for example, the following inventions can be provided without particular limitation.

[0011] (1) A method for promoting hatching of nematodes, comprising contacting an effective concentration of a hatching-promoting substance with eggs (in the case of nematodes forming cysts, cysts) of the nematodes, the hatching-promoting substance preferably comprising one or more selected from the group consisting of metal ions, salts of metal ions, oxidized metals, alkaline earth metal ions, salts of alkaline earth metal ions, chelated metals, chelated alkaline earth metals, plant extracts, neurotransmission-related compounds, metabolism-regulation-related factors, amino acids, and sulfur compounds.

[0012] (2) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising metal ions, the metal ions being one or more selected from the group consisting of iron ions and zinc ions.

[0013] (3) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising chelated metals, the chelated metals being one or more selected from the group consisting of chelated iron and chelated zinc.

[0014] (4) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising plant extracts, the plant extracts being one or more extracts selected from the group consisting of tobacco extract, pepper extract, black pepper extract, sugar beet meal extract, and leaf extract of tomato.

[0015] (5) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising alkaline earth metals, the alkaline earth metals comprising calcium ions.

[0016] (6) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising neurotransmission-related compounds, the neurotransmission-related compounds comprising a-chaconine.

[0017] (7) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising metabolism-regulation-related factors, the metabolism-regulation-related factors being one or more selected from the group consisting of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-aminolevulinic acid, vitamin C, citric acid, a-ketoglutaric acid, and vitamin Bl.

[0018] (8) The method according to the above (1), wherein the hatching-promoting substance comprises amino acids, the amino acids being acidic amino acids.

[0019] (9) The method according to the above (1), wherein the hatching-promoting substance is a hatching-promoting substance comprising sulfur compounds, the sulfur compounds comprising iron alum.

[0020] (10) A composition for promoting hatching of cyst nematodes, comprising one or more selected from the group consisting of metal ions, salts of metal ions, oxidized metals, alkaline earth metal ions, salts of alkaline earth metal ions, chelated metals, chelated alkaline earth metals, plant extracts, neurotransmission-related compounds, metabolism-regulation-related factors, amino acids, and sulfur compounds.

[0021] (11) A method for improving hatching efficiency of cyst nematodes, comprising allowing water to permeate from the outside into cysts containing eggs of cyst nematodes.

[0022] (12) A method for promoting hatching of cyst nematodes, comprising: allowing moisture to permeate into cysts of cyst nematodes; and preferably contacting an effective concentration of a hatching-promoting substance with eggs of the cyst nematodes taken out from the cysts, the hatching-promoting substance preferably being a hatching-promoting substance comprising one or more selected from the group consisting of metal ions, salts of metal ions, oxidized metals, alkaline earth metal ions, salts of alkaline earth metal ions, chelated metals, chelated alkaline earth metals, plant extracts, neurotransmission-related compounds, metabolism-regulation-related factors, amino acids, and sulfur compounds.

[0023] (13) A method for screening a substance that promotes hatching of cyst nematodes, comprising: contacting a test substance with eggs of the cyst nematodes taken out from cysts of the cyst nematodes that have been permeated with moisture; and selecting a test substance that promotes hatching of nematodes derived from the eggs as compared with a negative control.

[0024] (14) An egg of a nematode, which is an egg of a nematode in a state ready for hatching, obtained from a cyst that has been permeated with water from the outside, and isolated from the cyst. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 The number of hatchings after immersing Gp cysts in a moist state in an aqueous zinc sulfate solution is shown. In Figures 1 to 16 , indicates p < 0.05 with respect to water.

[0026] Figure 2 The number of hatchings after immersing Gp cysts in a dry state in an aqueous zinc sulfate solution is shown.

[0027] Figure 3 The number of hatchings after immersing Gr cysts in an aqueous zinc sulfate solution is shown.

[0028] Figure 4 The number of hatchings after immersing Gp cysts in an aqueous zinc chloride solution is shown.

[0029] Figure 5 The number of hatchings after immersing Gr cysts in an aqueous zinc chloride solution is shown.

[0030] Figure 6 The number of hatches after Gp sporangia were immersed in an aqueous solution of ferric sulfate is shown.

[0031] Figure 7 The number of hatches after Gr sporangia were immersed in an aqueous solution of ferric sulfate is shown.

[0032] Figure 8 The number of hatches after Gp sporangia were immersed in a plant activator Menedael (registered trademark) containing ferric citrate is shown.

[0033] Figure 9 The number of hatches after Gr sporangia were immersed in a plant activator Menedael (registered trademark) containing ferric citrate is shown.

[0034] Figure 10 The number of hatches after Gp sporangia were immersed in an aqueous solution containing various extracts is shown.

[0035] Figure 11 The number of hatches after Gp sporangia were immersed in an aqueous solution of calcium chloride, zinc nitrate, or alum is shown.

[0036] Figure 12 The number of hatches after Gp sporangia were immersed in an aqueous solution containing a calcium chelate-containing Perform Ca (registered trademark) is shown.

[0037] Figure 13 The number of hatches after Gp sporangia were immersed in an aqueous solution containing α-ketocarboxylic acid is shown.

[0038] Figure 14 The number of hatches after Gp sporangia were immersed in an aqueous solution containing various metabolic-related factors (5-aminoimidazole-4-carboxamide riboside (AICAR), 5-aminolevulinic acid, vitamin C, citric acid, α-ketoglutaric acid, or vitamin B1) is shown.

[0039] Figure 15 The number of hatches after Gp sporangia were immersed in an aqueous solution containing aspartic acid is shown.

[0040] Figure 16 The number of hatches after Gp sporangia were immersed in an aqueous solution containing hot spring water (Yunohana) is shown.

[0041] Figure 17 The number of hatches after Gr sporangia were immersed in a water-cultured tomato root extract is shown.

[0042] Figure 18 The number of hatches after Gp sporangia were immersed in a water-cultured tomato root extract is shown. DETAILED DESCRIPTION

[0043] In this specification, "nematodes" is a general term for animals belonging to the phylum Nematoda. Nematodes include non-parasitic nematodes and parasitic nematodes. Parasitic nematodes targeting plants cause damage to crops. In plant parasitic nematodes (Nematology), research on parasitic nematodes targeting plants is conducted. In this specification, "plant parasitic nematodes" refer to parasitic nematodes targeting plants. Among the nematodes, cyst-forming nematodes (nematodes of the family Heteroderae; also called "cyst nematodes") are found. Soybean cyst nematode ( Heterodera glycines ), beet nematode ( Heterodara schachtii ), potato cyst nematode ( Globodera rostochiensis ), Clover cyst nematode ( Heterodera trifolii ), potato white cyst nematode ( Globodera pallida ) are cyst-forming nematodes. Potato cyst nematodes and white potato cyst nematodes selectively parasitize Chenopodiaceae and Solanaceae plants (e.g., peppers, potatoes, tomatoes, etc.). Soybean cyst nematodes selectively parasitize legumes such as soybeans, adzuki beans, and common beans. Clover cyst nematodes parasitize Dianthus plants such as carnations.

[0044] In this specification, "cysts" refer to cysts containing eggs. Female cyst nematodes die with hundreds of eggs inside, and their bodies form cysts containing eggs. Cysts are generally resistant to pesticides and other chemicals.

[0045] In this specification, "effective concentration" is a concentration at which a hatching-promoting substance exhibits a desired effect. The desired effect of a hatching-promoting substance is typically an effect of promoting the hatching of plant-parasitic nematodes (particularly cyst nematodes) from cysts.

[0046] In this specification, "hatching" refers to the hatching of nematodes from eggs. Hatching can be hatching from eggs in cysts or hatching from eggs outside cysts.

[0047] In this specification, "hatching promoting substance" refers to a substance that promotes the hatching of nematode eggs compared to a negative control (such as water, distilled water or pure water). The hatching promoting substance can be a substance that promotes the hatching of nematode eggs outside the cyst compared to a negative control (such as water, distilled water or pure water). The hatching promoting substance can preferably be a substance that promotes the hatching of nematode eggs derived from cysts (or inside cysts) compared to a negative control (such as physiological saline, water, distilled water or pure water). The hatching promoting substance is used in soil. The soil can be farmland, in particular dry land (preferably dry land of Solanaceae plants such as potato dry land, tomato dry land, and pepper dry land). Typically, the hatching promoting substance is synthesized or isolated. The hatching promoting substance can be a substance that promotes the hatching of nematode eggs outside the cyst more strongly than a positive object (for example, any hatching promoting substance described in this specification).

[0048] According to the present disclosure, there is provided a method of promoting hatching of nematodes, which comprises bringing an effective concentration of a hatching-promoting substance into contact with eggs of the nematodes described above. According to the present disclosure, there is also provided a method of promoting hatching of nematodes, which comprises bringing an effective concentration of a hatching-promoting substance into contact with cysts of the nematodes described above. According to the method of the present disclosure, by the above-described contact with the hatching-promoting substance, it is possible to hatch eggs of nematodes (e.g., eggs contained in cysts). The method is performed, for example, in soil (e.g., soil of a farmland (e.g., upland field)). The soil in which the method of the present disclosure is implemented can be soil containing cysts of nematodes or soil judged to contain cysts of nematodes. As described later, if water is allowed to permeate from the outside of the cysts to the cysts of nematodes, it is possible to promote hatching of eggs contained in the cysts of nematodes. Therefore, the method can be performed in soil during a period in which the soil is moist due to rain or water (e.g., within 1 day, within 2 days, or within 3 days from the day of rain).

[0049] The hatching-promoting substance can be one or more selected from the group consisting of a metal ion, a salt of a metal ion, an oxidized metal, a chelated metal, a chelating agent, a plant extract, a fertilizer, a neurotransmission-related compound, a metabolism-regulation-related factor, an amino acid, and a sulfur compound.

[0050] As the metal ion, for example, iron ions and zinc ions can be listed. The metal ion can be in an ionized form or in a form of a salt of a metal ion.

[0051] As the salt of a metal ion, inorganic salts and organic salts of a metal ion can be listed. As the inorganic salt of a metal ion, for example, sulfate salts, ammonium sulfate salts (molybdate salts), bisulfate salts, chloride salts, bromide salts, iodide salts, nitrate salts, phosphate salts, carbonate salts, bicarbonate salts, perchlorate salts, hydroxide salts, hydroxyoxide salts, and the like can be listed. In addition, as the organic salt of iron ions, for example, formate salts, acetate salts, propionate salts, butyrate salts, oxalate salts, acrylate salts, methacrylate salts, citrate salts, ammonium citrate salts, pyrophosphate salts, p-toluenesulfonate salts, and the like can be listed. The salt of a metal ion can be only one, or a combination or mixture of two or more.

[0052] As the oxidized metal, for example, iron oxide and zinc oxide can be listed. The iron oxide can preferably be ferrous oxide.

[0053] A chelated metal is a metal in a state of being chelated with a chelating agent. By being chelated with a chelating agent, a metal can improve water solubility or improve absorbability to cells. As a chelating agent, there is no particular limitation as long as it can chelate a metal or a metal ion, and for example, humic acid, ethylenediaminetetraacetic acid, ethylenediamine diacetic acid, nitrilotriacetic acid, glutamic acid-N,N-diacetic acid, aspartic acid-N,N-diacetic acid, glycine, alanine, acetic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, salicylic acid, citric acid, lactic acid, glycolic acid, malic acid, tartaric acid, mandelic acid, suberic acid, sebacic acid, decanedioic acid, phosphoric acid, pyrophosphoric acid, hexametaphosphoric acid, hexylphosphonic acid, octylphosphonic acid, nitrilotriacetic acid, 1-hydroxyethane-1,1-diphosphonic acid, 1-hydroxyethane-1,1-diphosphonic acid, thiodiglycolic acid, thiodipropionic acid, thiodibutyric acid, propyl dithiodiglycolic acid, butyl dithiodiglycolic acid, butyl dithiodipropionic acid, ethyl dithiodibutyric acid, glycol ether diamine tetraacetic acid, 1,10-phenanthroline, siderophores (e.g., catechol-type siderophores, hydroxamate-type siderophores, etc.), and the like, and salts thereof can be exemplified. The metal chelated can be, for example, a metal described in the above explanation. The chelated metal can be composed of one kind, or a combination or mixture of two or more kinds.

[0054] The alkaline earth metal can be, for example, calcium or magnesium. The alkaline earth metal ion can be a calcium ion or a magnesium ion. The alkaline earth metal ion can be in an ionized state or in the form of a salt. The salt can be one or more selected from the group consisting of the above-described organic salts and inorganic salts. In addition, the chelating agent contained in the chelated alkaline earth metal can be one or more selected from the above-described chelating agents.

[0055] The plant extract is a fraction obtained by extracting the whole or a part of a plant. The plant can be, for example, a Solanaceae plant such as a potato, a tomato, an eggplant, or the like. In addition, the plant can be, for example, one or more selected from the group consisting of Nicotiana (e.g., tobacco), Capsicum (e.g., pepper), Piper (pepper), Beta (e.g., sugar beet pulp), and Solanum (e.g., tomato). The part of the plant can be one or more parts selected from the group consisting of a fruit, a stem, a leaf, and a root of the plant. The plant extract can be, for example, one or more selected from the group consisting of a leaf of tobacco, a fruit of pepper, a fruit of black pepper, sugar beet pulp, and a leaf of tomato.

[0056] The neurotransmission-related compound can be, for example, a-chaconine.

[0057] The metabolism-regulating-related substance may, for example, be one or more selected from the group consisting of vitamins (e.g., vitamin B1, vitamin C, etc.), nucleic acids (e.g., ribonucleotides, deoxyribonucleotides, such as 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR)), intermediates of glycolysis (e.g., glucose, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, dihydroxyacetone phosphate, glyceraldehyde-3-phosphate, 1,3-diphosphoglycerate, 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, and pyruvate), and intermediates of the citric acid cycle (e.g., oxaloacetate, citrate, aconitate, D-isocitrate, alpha-ketoglutarate, succinyl CoA, succinate, fumarate, and L-malate).

[0058] The amino acid may, for example, be one or more selected from the group consisting of isoleucine, leucine, lysine, methionine, phenylalanine, threonine (serine), tryptophan, valine, histidine, tyrosine, cysteine, aspartic acid, asparagine, serine, glutamic acid, glutamine, proline, glycine, alanine, and arginine. The amino acid may, for example, be an acidic amino acid, such as aspartic acid and glutamic acid.

[0059] The sulfur compound may, for example, be iron alum. The sulfur compound is contained in sinter, and a form of the sulfur compound contained in sinter can be used.

[0060] The composition of the present disclosure containing the hatching-promoting substance can further contain a substance that improves the sustained-release property of the hatching-promoting substance. As the substance that improves the sustained-release property of the hatching-promoting substance, for example, biodegradable polymers (e.g., polymers from organisms, such as humic acid, humin, etc.), natural resins, porous substances, synthetic resins, and inorganic carriers can be exemplified.

[0061] According to the present disclosure, the waste liquid of hydroponics of a plant of the Solanaceae family (e.g., including root exudates) can promote the hatching of nematodes derived from eggs in cysts. According to the present disclosure, the waste liquid of hydroponics of a plant of the Solanaceae family (e.g., including root exudates) can also be added to the hatching-promoting substance of the present disclosure to promote the hatching of cyst nematodes. Specifically, the waste liquid of hydroponics of tomato or the waste liquid of recirculating hydroponics of tomato (e.g., including root exudates) can be combined in the hatching-promoting substance of the present disclosure to promote the hatching of cyst nematodes. According to the present disclosure, the hatching of cyst nematodes in the soil of a dry field where a plant of the Solanaceae family (e.g., other than potato) is cultivated (e.g., including root exudates) can also be promoted by the hatching-promoting substance of the present disclosure, and preferably the hatching of cyst nematodes on rainy days can also be promoted by the hatching-promoting substance of the present disclosure. In addition, the hatching of cyst nematodes can also be promoted by contacting the cysts with a solution in which the waste liquid of hydroponics of a plant of the Solanaceae family is used as a solvent for the hatching-promoting substance of the present disclosure.

[0062] According to the present disclosure, there is provided a method of permeating water to cysts of cyst nematodes. The method of the present disclosure includes contacting or immersing the cysts of cyst nematodes in a sufficient amount of water for a sufficient time. In the method, the permeation of water to the cysts can change the cyst nematodes to a state in which the eggs contained in the cysts are easily hatched, and / or can increase the sensitivity of the eggs of the cyst nematodes to a hatching promoting substance. Thus, the method can be performed as a pre-stage of the cyst treatment based on the hatching promoting substance, or can be a pretreatment for the cyst treatment based on the hatching promoting substance. The water is not particularly limited, and can be distilled water. The method is performed, for example, in a laboratory or in vitro (for example, in a test container such as a test tube or a microtube).

[0063] The water may, for example, have a conductivity of 150 μS / cm or less, 100 μS / cm or less, 50 μS / cm or less, 10 μS / cm or less, 5 μS / cm or less, 4 μS / cm or less, 3 μS / cm or less, 2 μS / cm or less, 1 μS / cm or less, or 0.054 μS / cm or more. The water may, for example, have a conductivity of 5 μS / cm to 50 μS / cm. The water may, for example, have a conductivity of 0.054 μS / cm to 2 μS / cm. The conductivity can be measured at 25°C under atmospheric pressure.

[0064] According to the present disclosure, there is provided a method of promoting hatching of cyst nematodes, which includes permeating water to cysts of cyst nematodes, and contacting an effective concentration of a hatching promoting substance to eggs of the cyst nematodes taken out from the cysts. According to the present disclosure, there is provided a hatching promoting substance for use in such a method.

[0065] According to the present disclosure, there is provided a method of screening a hatching promoting substance for cyst nematodes, which includes providing a cyst permeated with water, and contacting a candidate for a hatching promoting substance to eggs taken out from the cyst permeated with water.

[0066] The cyst permeated with water can be obtained by contacting water under conditions suitable for the cysts of cyst nematodes.

[0067] It is considered that the eggs of the cyst nematode acquire resistance to the hatching promoting substance through abrupt contact with water. In contrast, according to the present disclosure, the eggs taken out from the cysts infiltrated with water have high reactivity to the hatching promoting substance compared to the eggs taken out from the cysts before infiltration with water. It is considered that by infiltrating water from the outside of the cyst into the cyst, abrupt contact of the eggs with water can be prevented, the eggs can be brought into a state easy to hatch, and acquisition of resistance to the hatching promoting substance can be prevented. Dispensing of the eggs of the nematode into the wells is easier than dispensing of the cysts. Generally, dispensing of the cysts requires moving one by one with forceps into the wells to make the number of eggs constant, in contrast, the eggs are dispensed into the wells in a constant number by dispensing a constant amount of the suspension thereof into the wells at a time. Therefore, the eggs taken out from the cysts infiltrated with water are advantageous in screening of the hatching promoting substance. In the present disclosure, the eggs of the nematode (preferably isolated eggs) in the state easy to hatch obtained from the cyst infiltrated with water from the outside and an aqueous solution containing the same are provided. The candidate of the hatching promoting substance can be, for example, a natural product, an extract of a natural product, a plant, a part thereof, an extract of a plant or a part thereof, a plant-derived component (protein, lipid, fiber, cell, nucleic acid, etc.). The candidate of the hatching promoting substance can be one or more selected from the group consisting of a metal ion, a salt of a metal ion, an oxidized metal, an alkaline earth metal ion, a salt of an alkaline earth metal ion, a chelated metal, a chelated alkaline earth metal, a plant extract, a neurotransmission-related compound, a metabolism-regulation-related factor, an amino acid, and a sulfur compound.

[0068] The method of screening according to the present disclosure can further include selecting a test substance that promotes hatching of the nematode derived from the eggs compared to a negative subject. As the negative subject, physiological saline, water, distilled water, and pure water can be exemplified.

[0069] According to the present disclosure, a hatching promoting composition is provided, which contains one or more selected from the group consisting of a metal ion, a salt of a metal ion, an oxidized metal, an alkaline earth metal ion, a salt of an alkaline earth metal ion, a chelated metal, a chelated alkaline earth metal, a plant extract, a neurotransmission-related compound, a metabolism-regulation-related factor, an amino acid, and a sulfur compound. The hatching promoting composition promotes hatching of the nematode derived from the cysts or eggs of the cyst nematode (particularly, Globodera pallida and Globodera rostochiensis).

[0070] According to the present disclosure, there is provided a method for improving soil contaminated with cysts or eggs of a cyst nematode (or a method for reducing the amount of cysts from the above-mentioned soil), which comprises: bringing an effective amount of any one or more of the above-mentioned hatching promoting substances into contact with the cysts or eggs of the cyst nematode; and thereby hatching the cyst nematode from the cysts or eggs. The method of the present disclosure can be carried out in the absence of a plant that is able to parasitize the cyst nematode. In the method of the present disclosure, the cysts can be permeated with water, or can have been dried. The method of the present disclosure can further comprise confirming the hatching of the cyst nematode. The method of the present disclosure can further comprise confirming the death of the cyst nematode. The removal of the cysts of the cyst nematode from the soil refers to the improvement of the soil. By the improvement of the soil, for example, a Solanaceae plant can be cultivated well in the soil without being harmed by the cyst nematode. As the harm caused by the parasitization of the cyst nematode, there can be cited the weakening of the growth of the plant (chlorosis) and the death. A plant parasitized by the cyst nematode can produce one or more selected from the group consisting of the reduction of the root system, water stress, and nutrient deficiency, and sometimes causes the aging of the plant body, the increase in the susceptibility to fungal infection, and the like. The method of the present disclosure can further comprise determining that the soil contains the cysts before the contact of the hatching promoting substance with the soil, or can not comprise it. According to the present disclosure, there is provided a hatching promoting substance for use in such a method.

[0071] According to the present disclosure, there is provided a method for disposing of an agricultural product contaminated with cysts or eggs of a cyst nematode (for example, for Globodera rostochiensis and Globodera pallida, a harvest from a Solanaceae plant, for Globodera solani, a harvest from a Leguminosae plant (Leguminosae plants such as soybean, azuki bean, and kidney bean), for Globodera tabacum, a Beta vulgaris or Brassica plant such as sugar beet, for Globodera rapae, a Dianthus plant such as carnation), which comprises: bringing an effective amount of any one or more of the above-mentioned hatching promoting substances into contact with the above-mentioned agricultural product; and thereby hatching the cyst nematode from the cysts or eggs. The method of the present disclosure can further comprise washing the agricultural product or bringing it into contact with a pesticide. The agricultural product is, for example, a crop. According to the present disclosure, there is provided a hatching promoting substance for use in such a method.

[0072] According to the present disclosure, there is provided a method of cultivating a crop, which includes: bringing a hatching promoting substance into contact with soil contaminated with cysts or eggs of a cyst nematode; and cultivating a crop using the soil. By bringing a hatching promoting substance into contact with soil contaminated with cysts or eggs of a cyst nematode, it is possible to hatch the cyst nematode. Here, the crop may, for example, be a crop of a kind that is not parasitized. Even if the cyst nematode hatches, there is no crop in the vicinity that can be parasitized, and thus the cyst nematode starves to death. By doing so, it is possible to increase the likelihood that there is no cyst nematode in the soil at the time of cultivation of a crop that can be parasitized later, or to reduce the amount of cyst nematode in the soil, or to remove the cyst nematode, for example, to the extent that no damage to the crop occurs. According to the present disclosure, there is provided a hatching promoting substance for use in such a method.

[0073] According to the present disclosure, there is provided a method of cultivating a crop, which includes: bringing a hatching promoting substance into contact with soil contaminated with cysts or eggs of a cyst nematode; bringing the hatched cyst nematode to death in the presence of a first crop of a kind that is not parasitized, or in the absence of a crop or the first crop; and cultivating a second crop after confirming the death of the cyst nematode. The second crop can be a plant of a kind that is parasitized by the cyst nematode, or a plant of a kind that is not parasitized. According to the present disclosure, there is provided a hatching promoting substance for use in such a method.

[0074] Embodiments

[0075] Example 1: Effect of test substances on hatching promotion of nematode cysts

[0076] Cyst nematodes hibernate in the form of cysts that are highly resistant to stress in the soil, hatch in response to a crop, parasitize or infect the crop, and cause damage to the crop. If the crop or its roots withers, it survives in the form of cysts in the soil.

[0077] As a positive control, the hatching efficiency of the nematodes was evaluated using tomato root leachate. The tomato root leachate was prepared as follows. That is, tomato seedlings were grown in a greenhouse for 2 weeks, and then the roots were collected and washed with distilled water. The roots were then crushed, and the crushed roots were filtered through a 100-mesh filter to obtain the tomato root leachate. Solanum lycopesicumThe variety "Home Momotaro" (registered trademark; Takii Seed) was planted in a pot (1 / 5000 ares) and cultivated for 30 days. A rubber plug was inserted into the pot and 1 L of distilled water was poured in. After 3 hours, the plug was removed and the waste water was recovered. The waste water was filtered with filter paper (No. 101, ADVANTEC Toyo Kaisha) and the filtrate was recovered. The filtrate was further pressure-filtered with a needle filter (PES, pore size 0.45 μm, Hawachi Scientific). This final filtrate was used as the tomato root extract. The tomato root extract was dispensed into a microtube and stored frozen at -20°C until use. Eggs of the cyst nematode were prepared. For the eggs, the cysts were ground on a sieve having a mesh of 250 μm, then the inclusions were removed with a sieve having a mesh of 106 μm, and the eggs of the cyst nematode were recovered on a sieve having a mesh of 25 μm. Group 1: eggs, group 2: a mixture of eggs and the shells of cysts, group 3: damaged cysts, and group 4: cysts were immersed in the tomato root extract, and the hatching rate was measured. Thus, group 1 was 17.2%, group 2 was 9.4%, group 3 was 5.6%, and group 4 was approximately 100%. From the above results, it was understood that the shells of the cysts were not related to the decrease in the hatching rate.

[0078] Next, the cysts were immersed in distilled water for about 1 week, whereby the horizontal stabilizer was allowed to permeate into the cysts, and the resulting cysts and the eggs from the cysts were subjected to the same experiment as described above. Thus, the hatching rate from the cysts was about 80%, and the hatching rate from the eggs was also about 80%. It was found that the hatching rate of the eggs was increased by immersing the cysts in water. In the case where the cysts are used in the screening test, the cysts must be picked up with tweezers and the number of cysts in each test area must be made uniform, but in the case where the eggs separated from the cysts are used, the number of eggs in each test area can be made uniform by dispensing the suspension of the eggs. Therefore, the agent screening system using the eggs is more practical than the agent screening system using the cysts.

[0079] Next, in the present embodiment, substances that promote the hatching of the cyst nematode were explored.

[0080] As the cyst nematode, potato white cyst nematode (Gp) and potato cyst nematode (Gr) were used. The cysts were immersed in distilled water for 1 week before the start of the experiment. Four cyst nematode cysts were inoculated in each well of a 96-well plate (containing 50 μL of distilled water). As the negative control (control), distilled water was used, and as the positive object, the tomato root extract was used. The test substance was used after being dissolved in distilled water. The upper opening of each well was closed with a sealing strip, and left standing in an incubator at 20°C. The number of hatched larvae in each well was counted every 7 days.

[0081] As Figures 1 to 8As shown in FIGS. 1 1 (A) and (B), various substances such as metal ions, divalent ion chelating agents, plant extracts, fertilizers, neurotransmission-related compounds, metabolism-regulation-related factors, amino acids, and sinter promoted the hatching of the cyst nematodes.

[0082] Root exudates were recovered from soil- or water-cultured tomatoes. Cysts of the nematodes were immersed in the stock solution or diluted solution of the root exudates, and the number of hatched larvae from the eggs in the cysts was counted. Figure 17 The circulating water in FIG. 12 refers to the liquid fertilizer before being circulated in the water culture of the tomatoes. As shown in FIG. 12, the root exudates of the water-cultured tomatoes promoted the hatching of the nematodes. Figure 17 As shown in FIGS. 13 (A) and (B), the root exudates of the water-cultured tomatoes promoted the hatching of the nematodes. Figure 18 As shown in FIGS. 14 (A) and (B), the root exudates of the water-cultured tomatoes promoted the hatching of the nematodes.

Claims

1. A method for promoting the hatching of nematodes, comprising contacting an effective concentration of a hatching-promoting substance with eggs of the nematode (or cysts in the case of nematodes that form cysts), The incubation promoting substance comprises one or more selected from the group consisting of metal ions, metal ion salts, metal oxides, alkaline earth metal ions, alkaline earth metal ion salts, chelated metals, chelated alkaline earth metals, plant extracts, neurotransmission-related compounds, metabolism regulation-related factors, amino acids and sulfur compounds.

2. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing metal ions, and the metal ions are at least one selected from the group consisting of iron ions and zinc ions.

3. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing a chelated metal, and the chelated metal is at least one selected from the group consisting of chelated iron and chelated zinc.

4. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing a plant extract, and the plant extract is one or more extracts selected from the group consisting of tobacco extract, pepper extract, black pepper extract, beet pulp extract, and tomato leaf extract.

5. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing an alkaline earth metal, and the alkaline earth metal contains calcium ions.

6. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing a neurotransmission-related compound, and the neurotransmission-related compound contains α-chaconine.

7. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing a metabolism regulating related factor, and the metabolism regulating related factor is one or more selected from the group consisting of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR), 5-aminolevulinic acid, vitamin C, citric acid, α-ketoglutaric acid or vitamin B1.

8. The method according to claim 1, wherein The hatching promoting substance contains amino acids, which are acidic amino acids.

9. The method according to claim 1, wherein The hatching promoting substance is a hatching promoting substance containing a sulfur compound, and the sulfur compound contains iron alum.

10. A composition for promoting the hatching of cyst nematodes, comprising one or more selected from the group consisting of metal ions, salts of metal ions, metal oxides, alkaline earth metal ions, salts of alkaline earth metal ions, chelated metals, chelated alkaline earth metals, plant extracts, compounds related to neurotransmission, factors related to metabolic regulation, amino acids, and sulfur compounds.

11. A method for improving the hatching efficiency of cyst nematodes, comprising allowing water to penetrate from outside the cyst into the cyst containing eggs of the cyst nematode.

12. A method for promoting the hatching of cyst nematodes, comprising: Allows water to penetrate into the cysts of cyst nematodes; and contacting an effective concentration of a hatching-promoting substance with the eggs of the cyst nematode removed from the cyst, The hatching promoting substance is a hatching promoting substance comprising one or more selected from the group consisting of metal ions, salts of metal ions, metal oxides, alkaline earth metal ions, salts of alkaline earth metal ions, chelated metals, chelated alkaline earth metals, plant extracts, neurotransmission-related compounds, metabolism regulation-related factors, amino acids and sulfur compounds.

13. A method for screening a substance that promotes the hatching of cyst nematodes, comprising: contacting a test substance with eggs of a cyst nematode extracted from a cyst of the cyst nematode that has been permeated with water; as well as A test substance is selected that promotes the hatching of nematodes from eggs compared to negative test substances.

14. Nematode eggs, which are obtained from cysts permeated with water from the outside, separated from the cysts, and are in a state in which they are easily hatched.

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