Fungicidal compositions and their use in controlling fusarium diseases of crops

Abstract

Five fungicidal compositions and their use in controlling Fusarium diseases in crops are disclosed, belonging to the field of plant disease control technology. The active ingredients of the compositions include the compound ZJS178 having the structural formula shown in formula (I) and a triazole fungicide, or the compound ZJS178 and a succinate dehydrogenase inhibitor fungicide, or the compound ZJS178 and a pyrrole fungicide, or the compound ZJS178 and an imidazole fungicide, or the compound ZJS178 and a strobilurin fungicide. The compositions of the present invention have a multi-component synergistic effect, are free of cross-resistance, and can significantly improve fungicidal activity and control efficacy, are advantageous in resolving and delaying resistance in disease fungi, are safe for crops, and meet the requirements of reducing the amount of pesticide application and improving efficiency.

Description

[Technical Field]

[0001] The present invention relates to the field of plant disease control technology, and in particular to five fungicidal compositions and their use in controlling Fusarium diseases of crops. [Background technology]

[0002] Fusarium spp. is a species of fungus found worldwide that causes a variety of plant diseases, including root rot, stem rot, base rot, blossom rot, and ear rot, causing wilting and death of crops and affecting their yield and quality. Common Fusarium diseases in crops include rice seedling disease, wheat head blight, corn ear rot, cucurbit wilt, and root rot of various crops.

[0003] In recent years, due to changes in climate and cultivation methods, wheat head blight caused by the Fusarium graminearum complex has become a serious threat to the safe production of wheat. Unlike other crop diseases, the Fusarium head blight fungus produces mycotoxins, such as deoxynivalenol (also known as vomitoxin, DON) and zearalenone (ZEA), in infected wheat, posing a serious threat to human and animal safety. Therefore, this disease has been classified as a "Class 1 Crop Pest" by the Ministry of Agriculture and Rural Affairs. Furthermore, with the widespread use of wheat and corn in rotation, corn ear rot caused by the Fusarium graminearum complex has shown a serious trend, and the agriculture and grain purchasing departments of major provincial agricultural provinces are placing great importance on this issue.

[0004] In addition to Fusarium graminearum, rice seedling disease caused by the Fusarium fujikuroi species complex is also a global fungal disease. In China, with the widespread use of intensive seedling raising techniques such as mechanized transplanting and seedling tossing, the occurrence of seedling disease has become increasingly common, with local incidence rates reaching 100%, resulting in serious yield losses.

[0005] Fusarium can cause wilt diseases of various fruits and vegetables, such as banana wilt caused by Fusarium oxysporum, which causes large areas of bananas to wither, yellow, and die, resulting in huge losses for banana farmers and becoming a "cancer" for the banana industry.Furthermore, wilt diseases of cucumbers, tomatoes, etc. caused by Fusarium oxysporum significantly restrict the healthy development of the industry.

[0006] Therefore, continuous and efficient prevention and control of Fusarium diseases in crops is of great importance to ensure the effective supply and quality safety of agricultural products in China.

[0007] Currently, chemical control remains the primary means of preventing and controlling Fusarium diseases in crops, as there are no highly resistant crop varieties available.

[0008] Currently, triazole fungicides are widely used to control Fusarium diseases in crops. Triazole fungicides are sterol biosynthesis inhibitors that inhibit the synthesis of ergosterol by the pathogen, thereby altering membrane-associated cellular functions and thereby inhibiting and killing the fungus. For example, tebuconazole (chemical name: (RS)-1-p-chlorophenyl-4,4-dimethyl-3-(1H-1,2,4-triazol-1-ylmethyl)pentan-3-ol) can be used to control various crop diseases, such as rust, scab, powdery mildew, net blotch, and root rot. Metconazole (chemical name: 5-(4-chlorophenyl)-2,2-dimethyl-1-(1H-1,2,4-triazol-1-ylmethyl)cyclopentanol) can be used to control various diseases in cereals, wheat, and legumes. Both have excellent protective and curative effects, with long-lasting efficacy. However, long-term use of these pesticides can lead to resistance in Fusarium diseases, affecting their effectiveness.

[0009] Succinate dehydrogenase inhibitor (SDHI) fungicides such as pydiflumetofen can be used to control Fusarium head blight and other cereal diseases caused by Fusarium. Pydiflumetofen (chemical name: 3-(difluoromethyl)-N-methoxy-1-methyl-N-[(RS)-1-methyl-2-(2,4,6-trichlorophenyl)ethyl]pyrazole-4-carboxamide) is a new generation succinate dehydrogenase inhibitor (SDHI) developed by Syngenta. This fungicide acts on respiratory chain complex II, inhibiting ATP synthesis and inhibiting the growth and death of pathogenic fungi, ultimately achieving the goal of plant disease control. Penflufen (chemical name: 5-fluoro-1,3-dimethyl-N-[2-(4-methylpent-2-yl)phenyl]-1H-pyrazole-4-carboxamide) is an SDHI fungicide developed by Bayer. This fungicide acts on respiratory complex II, inhibiting ATP synthesis in pathogenic fungi, and has systemic, preventive, and curative effects. Currently, it is primarily used as a seed treatment; after seed treatment, the fungicide penetrates the germinating seeds and spreads throughout the plant via the xylem of the seedlings, providing a protective role. However, because succinate dehydrogenase inhibitor fungicides have a single site of action, the risk of resistance continues to increase as they are widely used.

[0010] Fludioxonil (chemical name: 4-(2,2-difluoro-1,3-benzodioxy-4-yl)pyrrole-3-nitrile), a pyrrole fungicide, is commonly used to control diseases such as Fusarium crown rot (wheat root rot), rice seedling disease, and tomato wilt. This fungicide inhibits fungal mycelial growth primarily by activating the histidine kinase signaling pathway of the pathogen. It can be used as a seed treatment to control a variety of seed-borne and soil-borne pathogens, including those of Alternaria, Fusarium, Helminthosporium, Rhizoctonia, and Penicillium. However, existing research shows that pathogenic fungi are susceptible to developing resistance to this drug, which can significantly affect its effectiveness (Oiki et al., 2022. Wide distribution of resistance to the fungicides fludioxonil and iprodione in Penicillium species. PLOS ONE 17 (1); Dowling et al., 2021. Characterization of high fludioxonil resistance in Botrytis cinerea isolates from calibrachoa flowers. Phytopathology, 111(3): pp.478-484; Wen et al., 2022. Biological and molecular characterizations of field fludioxonil-resistant isolates of Fusarium graminearum. Pesticide biochemistry and physiology, 184: DOI10.1016 / j.pestbp.2022.105101.).

[0011] Prochloraz (chemical name: N-propyl-N-[2-(2,4,6-trichlorophenoxy)ethyl]imidazole-1-carboxamide) is a broad-spectrum fungicide that exhibits clear control effects against various crop diseases caused by Ascomycetes and Deuteromycetes. Carbendazim (chemical name: benzimidazol-2-ylmethylcarbamate) is a broad-spectrum systemic fungicide that primarily interferes with the mitotic process and is effective against some Ascomycete pathogens and most Deuteromycete pathogens. Thiophanate-methyl (chemical name: 1,2-bis(3-methoxycarbonyl-2-thioureayl)benzene) is a broad-spectrum, systemic, and low-toxic fungicide with systemic and conductive properties. This drug acts on the beta-tubulin of pathogenic fungi, inhibiting cell division and suppressing the growth of the pathogens.

[0012] Azoxystrobin (chemical name: (E)-2-{2-[6-(2-cyanophenoxy)pyrimidin-4-yloxy]phenyl}-3-methoxyacrylate) is a strobilurin fungicide that inhibits electron transfer between fungal cytochromes bc1, thereby inhibiting mitochondrial respiration and causing fungal cell death. It has excellent control effects against a variety of crop diseases caused by fungal pathogens, including Ascomycota, Basidiomycota, Flagellates, and Imperfectimycota. Its highly effective and broad-spectrum action against powdery mildew, rust, leaf spot, net blotch, downy mildew, and blast on cereals, rice, peanuts, grapes, potatoes, fruit trees, vegetables, coffee, and lawns. It can be used as a foliar spray, seed treatment, or soil treatment. Trifloxystrobin (chemical name: (E)-methoxyimine-{(E)-α-[1-(α,α,α-trifluoro-m-methyl)ethyliminoxy]o-tolyl}acetate) is a broad-spectrum fungicide with long-lasting activity, capable of protecting, treating, and eradicating most diseases caused by pathogenic fungi, including Ascomycota, Basidiomycota, Oomycota, and Fungi Imperfecti, and has no cross-resistance with existing fungicides. Fluoxastrobin (chemical name: {2-[6-(2-chlorophenoxy)-5-fluoropyrimidin-4-yloxy]phenyl}(5,6-dihydro-1,4,2-dioxazin-3-yl)methyl ketone o-methyloxime) has broad-spectrum fungicidal activity and exhibits excellent activity against almost all fungal diseases (Ascomycota, Basidiomycota, Oomycota, and Deuteromycota), including rust, leaf spot, net blotch, powdery mildew, downy mildew, and other diseases. Picoxystrobin (chemical name: (E)-3-methoxy-2-{2-[6-(trifluoromethyl)-2-pyridyloxymethyl]phenyl}methyl acrylate) is a broad-spectrum systemic fungicide.Pyraclostrobin (chemical name: N-[2-[[1-(4-chlorophenyl)pyrazol-3-yl]oxymethyl]phenyl]-N-methoxycarbamate (N-[2-[[1-(4-chlorophenyl))-1H-pyrazol-3-yloxymethyl]phenyl](N-methoxy)carbamate)) has protective and therapeutic effects, permeability, and resistance to washaway by rainwater, making it suitable for a wide range of uses. Phenaminestrobin (chemical name: (E,E,E)-N-methyl-2-[((((1-methyl-3-(2,6-dichlorophenyl)-2-propenyl)imino)oxy)methyl)phenyl]-2-methoxyiminoacetamide) has a broad fungicidal spectrum, high activity, and both preventive and curative properties. It has good control effects against various plant diseases caused by flagellates, zygomycetes, ascomycetes, basidiomycetes, and imperfect fungi, and has excellent control effects against powdery mildew and rust.

[0013] Long-term use of pesticides can lead to the development of resistance in pathogens, which can affect the effectiveness of the pesticides. Therefore, the development of appropriate pesticide combinations is particularly important for the prevention and control of Fusarium diseases in crops. Summary of the Invention [Problem to be solved by the invention]

[0014] The object of the present invention is to provide a highly efficient, low-toxicity, and environmentally friendly fungicidal composition that is effective against crop diseases caused by Fusarium, and that can delay drug resistance and extend the useful life of the drug. [Means for solving the problem]

[0015] To achieve the above objectives, the present invention adopts the following technical solutions:

[0016] In a first aspect, the present invention provides a fungicidal composition comprising, as active ingredients, a compound ZJS178 having the structural formula shown in formula (I) and a triazole fungicide. [ka]

[0017] The above compound ZJS178 is a new compound synthesized in the early stage by the research group of the present invention. Its chemical name is 2-cyano-3-amino-3-[4-(N-ethyl-N-methylamino)phenyl]ethyl acrylate, and its synthetic route is as follows: [ka]

[0018] This compound is a cyanoacrylate fungicide that acts on Fusarium type I myosin (a motor protein) to cause the fungus to lose its growth motive force and die. Research has shown that ZJS178 has a new structure and unique mechanism of action, is not cross-resistant with triazole fungi, and is effective against diseases caused by Fusarium, providing both protective and curative effects.

[0019] Because ZJS178 and triazole fungicides have different mechanisms of action, the present invention combines the two to significantly improve the control effect against Fusarium disease in crops compared to when the two are used alone, reducing the application rate of the individual agent. This helps delay the development of drug resistance in pathogens, extend the service life of the fungicide, and reduce costs.

[0020] Preferably, the mass ratio of the compound ZJS178 to the triazole fungicide in the composition is 50:1 to 1:50.

[0021] The triazole fungicides include, but are not limited to, tebuconazole, metconazole, myclobutanil, difenoconazole, triadimefon, epoxiconazole, diniconazole, prothioconazole, propiconazole, and the like.

[0022] Preferably, the triazole fungicide is tebuconazole or metconazole.

[0023] More preferably, the mass ratio of the compound ZJS178 to the triazole fungicide in the composition is 15:1 to 1:15.

[0024] Research has shown that when compound ZJS178 is combined with a triazole fungicide at a certain ratio, a synergistic effect is observed, resulting in a synergistic effect in the control of Fusarium diseases. For example, in the case of Fusarium oxysporum, a synergistic effect is observed when compound ZJS178 is combined with a triazole fungicide at a mass ratio of 8:1 to 1:8. In the case of Fusarium fujikuroi, a synergistic effect is observed in the inhibition of metconazole-susceptible Fusarium fujikuroi when compound ZJS178 is combined with a triazole fungicide at a mass ratio of 4:1 to 1:5, and a synergistic effect is observed in the inhibition of metconazole-resistant Fusarium fujikuroi when compound ZJS178 is combined with a triazole fungicide at a mass ratio of 5:1 to 1:3. In the case of Fusarium graminearum, when compound ZJS178 is combined with a triazole fungicide in a mass ratio of 3:1 to 1:3, it has a synergistic effect on the inhibition of tebuconazole-susceptible Fusarium graminearum, and when combined with a mass ratio of 2:1 to 1:3, it has a synergistic effect on the inhibition of tebuconazole-resistant Fusarium graminearum.

[0025] Preferably, the mass ratio of the compound ZJS178 to the triazole fungicide in the composition is 8:1 to 1:8.

[0026] More preferably, the compound ZJS178 and the triazole fungicide are combined in a mass ratio of 2:1 to 1:3.

[0027] The fungicidal composition further contains auxiliary ingredients necessary for the formulation of a pesticide, and the active ingredient in the composition is 1 to 90% by mass. The auxiliary ingredients are carriers and adjuvants commonly used in pesticides.

[0028] Preferably, the mass percentage of the active ingredient in the composition is 15 to 40%.

[0029] According to methods known to those skilled in the art, the fungicidal compositions described in the present invention can be formulated into various agriculturally acceptable dosage forms for practical application, including water dispersible granules (water dispersible granules), suspensions, water emulsions, microemulsions, and seed treatment suspensions.

[0030] For the water dispersible granule formulation, those skilled in the art can use corresponding auxiliaries to complete the present invention. Polycarboxylates, lignin sulfonates, alkyl naphthalene sulfonates, etc. can be selected for use as dispersants; alkyl sulfates, alkyl sulfonates, naphthalene sulfonates can be selected for use as wetting agents; ammonium sulfate, urea, sucrose, and glucose can be selected for use as disintegrants; diatomaceous earth, corn starch, polyvinyl alcohol, and carboxymethyl (ethyl) celluloses can be selected for use as binders; and diatomaceous earth, kaolin, white carbon black, light calcium, talc powder, attapulgite, and china clay can be selected for use as fillers.

[0031] The auxiliary agents that can be used in suspension formulations are as follows: polycarboxylates, lignin sulfonates, and alkylnaphthalene sulfonates are selected as dispersants; alkylphenol polyoxyethylene ether formaldehyde condensate sulfates (alkylphenol ethoxylate formaldehyde condensate sulfates), alkylphenol polyoxyethylene ether phosphates (alkylphenol ethoxylate phosphates), phenylethylphenol polyoxyethylene ether phosphates (phenylethylphenol ethoxylate phosphates), alkyl sulfates, alkyl sulfonates, and naphthalene sulfonates are selected as wetting agents; xanthan gum, polyvinyl alcohol, and bentonite are selected as thickeners; formaldehyde, benzoic acid, and sodium benzoate are selected as preservatives; silicone-based antifoaming agents are selected as antifreezing agents; and inorganic salts such as ethylene glycol, propylene glycol, glycerin, urea, and sodium chloride are selected as antifreezing agents.

[0032] The following auxiliary agents can be used in water emulsion formulations: Emulsifiers include nonylphenol polyoxyethylene ether phosphate ester (nonylphenol ethoxylate phosphate ester), triphenylethylphenol polyoxyethylene ether phosphate ester (triphenylethylphenol ethoxylate phosphate ester) (Agricultural milk 600# phosphate ester), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, emulsifier T-60, surfactant TX-10, Agricultural milk 1601#, Agricultural milk 600#, Agricultural milk 400# is selected and used, xylene, toluene, cyclohexanone, oil solvents (solvent oil) (S-150, S-180, S-200) are selected and used as solvents, triphenyl phosphite, epichlorohydrin, acetic anhydride are selected and used as stabilizers, xanthan gum, polyvinyl alcohol, bentonite, magnesium aluminum silicate are selected and used as thickeners, and methanol, benzoic acid, sodium benzoate are selected and used as preservatives.

[0033] The following auxiliary agents can be used in microemulsion formulations: calcium dodecylbenzenesulfonate (Agricultural milk 500#), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, Tween-80-60#, TX-10, Agricultural milk 1601, Agricultural milk 600#, and Agricultural milk 400# are selected as emulsifiers; methanol, isopropyl alcohol, n-butanol, and ethanol are selected as co-emulsifiers; cyclohexanone, N-methylpyrrolidone, xylene, toluene, and oil solvents (S-150, S-180, and S-200) are selected as solvents; and triphenyl phosphite and epichlorohydrin are selected as stabilizers.

[0034] Adjuvants that can be used in seed treatment suspensions are: Film-forming agents may be polyethylene glycol, polyvinyl alcohol resins, polyvinylpyrrolidone, hydroxyethyl cellulose, homopolyvinyl acetate, sodium carboxymethylcellulose, gum arabic, gelatin, polyvinyl alcohol or polyacrylamide, or mixtures thereof. The technician can adjust the proportion of homopolyvinyl acetate depending on the required viscosity of the product.

[0035] The present invention also provides the use of the fungicidal composition in controlling crop diseases caused by Fusarium.

[0036] Furthermore, the Fusarium refers to plant pathogenic Fusarium, and mainly includes Fusarium graminearum complex, Fusarium fujikuroi complex, Fusarium oxysporum, and Fusarium moniliforme.

[0037] Furthermore, diseases of the above crops include wheat Fusarium head blight, rice seedling disease, and strawberry wilt (rice blast).

[0038] Specifically, when used to control wheat Fusarium head blight, the fungicide composition is applied at the heading stage and early flowering stage of wheat, and then applied again once after 6 to 7 days.

[0039] In a second aspect, the present invention provides an effective fungicidal composition, the active ingredient of which comprises a compound ZJS178 having the structural formula shown in formula (I) and a succinate dehydrogenase inhibitor fungicide. [ka]

[0040] The compound ZJS178 is a novel compound synthesized in the early stage by the research group of the present invention. Its chemical name is 2-cyano-3-amino-3-[4-(N-ethyl-N-methylamino)phenyl]ethyl acrylate, and its synthetic route is as follows: [ka]

[0041] This compound is a cyanoacrylate fungicidal compound that acts on Fusarium type I myosin, a motor protein in the pathogen that hydrolyzes ATP and converts bioenergy into mechanical energy, providing the driving force for the pathogen's vital activities, such as growth, infection, and toxin synthesis. ZJS178 specifically acts on Fusarium type I myosin, causing the pathogen to lose its growth motive power and killing the bacteria.

[0042] ZJS178 has a novel structure and unique mode of action. Research has shown that it has no cross-resistance with succinate dehydrogenase inhibitors (SDHIs) and is effective against diseases caused by Fusarium species, with protective and curative effects.

[0043] Although ZJS178 and SDHI fungicides have different mechanisms of action, their sites of action are related. Therefore, by combining the two, the present invention can significantly enhance fungicidal efficacy, delay the development of fungicide resistance in pathogenic bacteria, and reduce control costs. Specifically, SDHI fungicides can effectively inhibit ATP synthesis in pathogenic bacteria, and the appropriate combination of ZJS178 and SDHI fungicides can simultaneously block ATP synthesis and motor protein activity, which is equivalent to simultaneously blocking the "gasoline" and "engine" of pathogenic bacteria, thereby effectively inhibiting pathogenic bacterial growth and delaying the development of fungicide resistance to either of the fungicides.

[0044] Preferably, the mass ratio of the compound ZJS178 to the succinate dehydrogenase inhibitor fungicide in the composition is 50:1 to 1:50.

[0045] The succinate dehydrogenase inhibitor fungicides include, but are not limited to, pydiflumetofen, penflufen, fluxapyroxad, fluopyram, carboxin, and the like.

[0046] Preferably, the mass ratio of compound ZJS178 to succinate dehydrogenase inhibitor fungicide in the composition is 9:1 to 1:9.

[0047] Research has shown that when compound ZJS178 is combined with a succinate dehydrogenase inhibitor fungicide in a certain ratio, a synergistic effect is generated, resulting in a synergistic effect in controlling Fusarium diseases.When the mass ratio of compound ZJS178 to succinate dehydrogenase inhibitor fungicide in the composition is 7:1 to 1:5, a synergistic effect is achieved in inhibiting Fusarium graminearum and Fusarium fujikuroi.

[0048] Furthermore, the effective fungicidal composition further comprises an agriculturally acceptable carrier and adjuvant, and the active ingredient in the composition is 1 to 90% by weight.

[0049] Preferably, the active ingredient in the composition is 10 to 40% by mass.

[0050] According to methods known to those skilled in the art, the fungicidal compositions described in the present invention can be formulated into various agriculturally acceptable dosage forms for practical application, and commonly used formulation forms include water dispersible granules, suspensions, microemulsions, water emulsions, suspoemulsions, and seed treatment suspensions.

[0051] For the water dispersible granule formulation, those skilled in the art can use corresponding auxiliaries to complete the present invention. Polycarboxylates, lignin sulfonates, alkyl naphthalene sulfonates, etc. can be selected for use as dispersants; alkyl sulfates, alkyl sulfonates, naphthalene sulfonates can be selected for use as wetting agents; ammonium sulfate, urea, sucrose, and glucose can be selected for use as disintegrants; diatomaceous earth, corn starch, polyvinyl alcohol, and carboxymethyl (ethyl) celluloses can be selected for use as binders; and diatomaceous earth, kaolin, white carbon black, light calcium, talc powder, attapulgite, and china clay can be selected for use as fillers.

[0052] The auxiliary agents that can be used in the suspension formulation are as follows: polycarboxylates, lignin sulfonates, and alkylnaphthalene sulfonates are selected as dispersants; alkylphenol polyoxyethylene ether formaldehyde condensate sulfates, alkylphenol polyoxyethylene ether phosphates, phenylethylphenol polyoxyethylene ether phosphates, alkyl sulfates, alkyl sulfonates, and naphthalene sulfonates are selected as wetting agents; xanthan gum, polyvinyl alcohol, and bentonite are selected as thickeners; formaldehyde, benzoic acid, and sodium benzoate are selected as preservatives; silicone-based antifoaming agents are selected as antifreezing agents; and inorganic salts such as ethylene glycol, propylene glycol, glycerin, urea, and sodium chloride are selected as antifreezing agents.

[0053] The following auxiliary agents can be used in water emulsion formulations: emulsifiers include nonylphenol polyoxyethylene ether phosphate ester, triphenylethylphenol polyoxyethylene ether phosphate ester (Agricultural milk 600# phosphate ester), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, emulsifier T-60, surfactant TX-10, Agricultural milk 1601#, Agricultural milk 600#, and Agricultural milk 400#; solvents include xylene, toluene, cyclohexanone, and oil solvents (S-150, S-180, and S-200); stabilizers include triphenyl phosphite, epichlorohydrin, and acetic anhydride; thickeners include xanthan gum, polyvinyl alcohol, bentonite, and magnesium aluminum silicate; and preservatives include methanol, benzoic acid, and sodium benzoate.

[0054] The following auxiliary agents can be used in microemulsion formulations: calcium dodecylbenzenesulfonate (Agricultural milk 500#), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, Tween-80-60#, TX-10, Agricultural milk 1601, Agricultural milk 600#, and Agricultural milk 400# are selected as emulsifiers; methanol, isopropyl alcohol, n-butanol, and ethanol are selected as co-emulsifiers; cyclohexanone, N-methylpyrrolidone, xylene, toluene, and oil solvents (S-150, S-180, and S-200) are selected as solvents; and triphenyl phosphite and epichlorohydrin are selected as stabilizers.

[0055] Adjuvants that can be used in seed treatment suspensions are: Film-forming agents may be polyethylene glycol, polyvinyl alcohol resins, polyvinylpyrrolidone, hydroxyethyl cellulose, homopolyvinyl acetate, sodium carboxymethylcellulose, gum arabic, gelatin, polyvinyl alcohol or polyacrylamide, or mixtures thereof. The technician can adjust the proportion of homopolyvinyl acetate depending on the required viscosity of the product.

[0056] The present invention also provides for the use of an effective fungicidal composition in controlling Fusarium diseases in crops.

[0057] Furthermore, the Fusarium refers to plant pathogenic Fusarium, and mainly includes Fusarium graminearum complex, Fusarium fujikuroi complex, Fusarium oxysporum, and Fusarium moniliforme.

[0058] Additionally, crop diseases caused by Fusarium include, but are not limited to, wheat head blight and rice seedling disease.

[0059] Specifically, when a combination of ZJS178 and pydiflumetofen is used to control wheat Fusarium head blight, it is applied once at the heading stage and early flowering stage of wheat, and then again once six days later. The combination of ZJS178 and penflufen is used as a seed treatment agent to control crop diseases.

[0060] In a third aspect, the present invention provides a fungicide composition comprising as an active ingredient a compound ZJS178 having the structural formula shown in formula (I) and a pyrrole fungicide. [ka]

[0061] The above compound ZJS178 is a new compound synthesized in the early stage by the research group of the present invention. Its chemical name is 2-cyano-3-amino-3-[4-(N-ethyl-N-methylamino)phenyl]ethyl acrylate, and its synthetic route is as follows: [ka]

[0062] This compound is a cyanoacrylate fungicidal compound that acts on Fusarium type I myosin (a motor protein), causing the pathogen to lose its growth motive force and die.

[0063] Compound ZJS178 has a novel structure and unique mode of action, and is not cross-resistant with pyrrole fungicides such as fludioxonil. Because ZJS178 and pyrrole fungicides have different mechanisms of action, the present invention combines these two compounds to not only expand the fungicidal spectrum and enhance the fungicidal effect, but also delay the development of fungicide resistance in pathogens.

[0064] Preferably, the mass ratio of the compound ZJS178 to the pyrrole fungicide in the composition is 50:1 to 1:50.

[0065] The pyrrole fungicides include, but are not limited to, fludioxonil, fenpiclonil, and the like.

[0066] More preferably, the mass ratio of the compound ZJS178 to the pyrrole fungicide is 11:1 to 1:11.

[0067] Research has shown that when compound ZJS178 is combined with pyrrole fungicides in a certain ratio, a synergistic effect occurs, and thus the two have a synergistic effect on controlling Fusarium diseases. Specifically, in the case of Fusarium pseudograminearum, a combination of ZJS178 and a pyrrole fungicide in a mass ratio of 3:1 to 1:2 has a synergistic effect in inhibiting fludioxonil-resistant Fusarium pseudograminearum, and a combination of the two in a mass ratio of 1:2 to 3 has a synergistic effect in inhibiting fludioxonil-susceptible Fusarium pseudograminearum. Against Fusarium fujikuroi, a combination of ZJS178 and a pyrrole fungicide in a mass ratio of 1:1 to 2 has a synergistic effect. Against Fusarium oxysporum, a combination of ZJS178 and a pyrrole fungicide in a mass ratio of 1:1 to 3 has a synergistic effect.

[0068] Preferably, the mass ratio of the compound ZJS178 to the pyrrole fungicide in the fungicide composition is 1:1-3.

[0069] Furthermore, the composition further contains, in addition to the above active ingredient, an agriculturally acceptable carrier and adjuvant, provided that the active ingredient is present in an amount of 1 to 90% by mass.

[0070] Preferably, the active ingredient in the composition is 25 to 40% by mass.

[0071] For practical application, the fungicidal compositions described in the present invention can be formulated into various agriculturally acceptable dosage forms according to methods known to those skilled in the art, with preferred formulation forms including seed treatment suspensions, water dispersible granules, microemulsions, and water emulsions.

[0072] For seed treatment suspensions, those skilled in the art are well aware that the present invention can be accomplished using corresponding auxiliaries. The auxiliaries that can be used in the seed treatment suspensions are as follows: The film-forming agent may be polyethylene glycol, polyvinyl alcohol resin, polyvinylpyrrolidone, hydroxyethyl cellulose, homopolyvinyl acetate, sodium carboxymethylcellulose, gum arabic, gelatin, polyvinyl alcohol, or polyacrylamide, or a mixture thereof. The technician can adjust the proportion of homopolyvinyl acetate according to the required viscosity of the product.

[0073] For the water dispersible granule formulation, polycarboxylates, lignin sulfonates, alkyl naphthalene sulfonates, etc. can be selected and used as dispersants; alkyl sulfates, alkyl sulfonates, naphthalene sulfonates can be selected and used as wetting agents; ammonium sulfate, urea, sucrose, and glucose can be selected and used as disintegrants; diatomaceous earth, corn starch, polyvinyl alcohol, and carboxymethyl (ethyl) celluloses can be selected and used as binders; and diatomaceous earth, kaolin, white carbon black, light calcium, talc powder, attapulgite, and china clay can be selected and used as fillers.

[0074] The following auxiliary agents can be used in water emulsion formulations: emulsifiers include nonylphenol polyoxyethylene ether phosphate ester, triphenylethylphenol polyoxyethylene ether phosphate ester (Agricultural milk 600# phosphate ester), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, emulsifier T-60, surfactant TX-10, Agricultural milk 1601#, Agricultural milk 600#, and Agricultural milk 400#; solvents include xylene, toluene, cyclohexanone, and oil solvents (S-150, S-180, and S-200); stabilizers include triphenyl phosphite, epichlorohydrin, and acetic anhydride; thickeners include xanthan gum, polyvinyl alcohol, bentonite, and magnesium aluminum silicate; and preservatives include methanol, benzoic acid, and sodium benzoate.

[0075] The following auxiliary agents can be used in microemulsion formulations: calcium dodecylbenzenesulfonate (Agricultural milk 500#), Agricultural milk 700#, Agricultural milk 2201#, Span-60#, Tween-80-60#, TX-10, Agricultural milk 1601, Agricultural milk 600#, and Agricultural milk 400# are used as emulsifiers; methanol, isopropyl alcohol, n-butanol, and ethanol are used as co-emulsifiers; cyclohexanone, N-methylpyrrolidone, xylene, toluene, and oil solvents (grades (brands): S-150, S-180, and S-200) are used as solvents; and triphenyl phosphite and epichlorohydrin are used as stabilizers.

[0076] The present invention also provides the use of the fungicide composition in controlling crop diseases caused by Fusarium.

[0077] Furthermore, the Fusarium refers to plant pathogenic Fusarium, and mainly includes Fusarium pseudograminearum, Fusarium fujikuroi, Fusarium oxysporum, etc.

[0078] The crop diseases include, but are not limited to, wheat root rot, rice seedling disease, and tomato wilt.

[0079] Furthermore, the use includes preparing the fungicide composition into a seed treatment agent and treating the seeds of crops.

[0080] In a fourth aspect, the present invention provides a fungicidal composition comprising a compound ZJS178 having the structure of formula (I) and an imidazole fungicide. [ka]

[0081] The above compound ZJS178 is a new compound synthesized in the early stage by the research group of the present invention. Its chemical name is 2-cyano-3-amino-3-[4-(N-ethyl-N-methylamino)phenyl]ethyl acrylate, and its synthetic route is as follows: [ka]

[0082] This compound is a cyanoacrylate fungicide that acts on Fusarium type I myosin (a motor protein) to cause the fungus to lose its growth motive force and die. ZJS178 has a new structure and unique mode of action, and is not cross-resistant with imidazole fungicides.

[0083] Preferably, the mass ratio of ZJS178 to the imidazole fungicide in the composition is 60:1 to 1:60.

[0084] The imidazole fungicides include, but are not limited to, prochloraz, carbendazim, prochloraz-manganese chloride complex, thiophanate methyl, and the like.

[0085] More preferably, the mass blending ratio of ZJS178 to the imidazole fungicide in the composition is 20:1 to 1:20.

[0086] More preferably, the mass blending ratio of ZJS178 to the imidazole fungicide in the composition is 12:1 to 1:12.

[0087] More preferably, the mass ratio of ZJS178 to the imidazole fungicide in the composition is 6:1 to 1:6.

[0088] Most preferably, the mass ratio of ZJS178 to the imidazole fungicide in the composition is 3:1 to 1:3.

[0089] The compositions according to the invention are suitable for disinfection, in particular for agricultural disinfection.

[0090] Preferably, the compositions according to the invention are suitable for controlling diseases caused by Fusarium.

[0091] More preferably, the Fusarium refers to plant pathogenic Fusarium, mainly including Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, and Fusarium moniliforme.

[0092] More preferably, crop diseases caused by Fusarium include, but are not limited to, cucurbit wilt, tomato wilt, banana wilt, cotton wilt, strawberry wilt, rice seedling disease, and wheat head blight.

[0093] The disinfectant according to the present invention contains the composition in an amount of 0.5 to 90% by weight.

[0094] More preferably, the disinfectant contains the composition in an amount of 5 to 80% by weight.

[0095] The fungicide according to the present invention may further comprise, in addition to the composition, agriculturally acceptable carriers and adjuvants.

[0096] Because ZJS178 and imidazole fungicides have different mechanisms of action, the combination of the two in this invention provides significantly higher control effects against diseases caused by Fusarium than when used alone, reducing the application rate of the individual agents, delaying the development of drug resistance in pathogens, and helping to extend the service life of the fungicide.

[0097] Research has shown that when compound ZJS178 is combined with imidazole fungicides in a certain ratio, it has a synergistic effect in inhibiting the growth of Fusarium. For example, when compound ZJS178 is combined with thiophanate-methyl in a mass ratio of 3:1 to 1:3, it has a significant synergistic effect in inhibiting the growth of Fusarium oxysporum.

[0098] Furthermore, the fungicide further contains auxiliary ingredients necessary for the formulation of an agricultural chemical, and the active ingredient in the composition is 0.5 to 90 mass %. The auxiliary ingredients are carriers and adjuvants commonly used in agricultural chemicals.

[0099] According to methods known to those skilled in the art, the fungicidal compositions described in the present invention can be formulated into various agriculturally acceptable dosage forms, including water dispersible granules, suspensions, and seed treatment suspensions, for practical application.

[0100] For the water dispersible granule formulation, polycarboxylates, lignin sulfonates, alkyl naphthalene sulfonates, etc. can be selected and used as dispersants; alkyl sulfates, alkyl sulfonates, naphthalene sulfonates can be selected and used as wetting agents; ammonium sulfate, urea, sucrose, and glucose can be selected and used as disintegrants; diatomaceous earth, corn starch, polyvinyl alcohol, and carboxymethyl celluloses can be selected and used as binders; and diatomaceous earth, kaolin, white carbon black, light calcium, talc powder, attapulgite, china clay, etc. can be selected and used as fillers.

[0101] The auxiliary agents that can be used in the suspension formulation are as follows: polycarboxylates, lignin sulfonates, and alkylnaphthalene sulfonates are selected as dispersants; alkylphenol polyoxyethylene ether formaldehyde condensate sulfates, alkylphenol polyoxyethylene ether phosphates, phenylethylphenol polyoxyethylene ether phosphates, alkyl sulfates, alkyl sulfonates, and naphthalene sulfonates are selected as wetting agents; xanthan gum, polyvinyl alcohol, and bentonite are selected as thickeners; formaldehyde, benzoic acid, and sodium benzoate are selected as preservatives; silicone-based antifoaming agents are selected as antifreezing agents; and inorganic salts such as ethylene glycol, propylene glycol, glycerin, urea, and sodium chloride are selected as antifreezing agents.

[0102] Adjuvants that can be used in seed treatment suspensions are: Film-forming agents may be polyethylene glycol, polyvinyl alcohol resins, polyvinylpyrrolidone, hydroxyethyl cellulose, homopolyvinyl acetate, sodium carboxymethylcellulose, gum arabic, gelatin, polyvinyl alcohol or polyacrylamide, or mixtures thereof. The technician can adjust the proportion of homopolyvinyl acetate depending on the required viscosity of the product.

[0103] In a fifth aspect, the present invention provides a fungicidal composition comprising a compound ZJS178 having the structure of formula (I) and a strobilurin fungicide. [ka]

[0104] The above compound ZJS178 is a new compound synthesized in the early stage by the research group of the present invention. Its chemical name is 2-cyano-3-amino-3-[4-(N-ethyl-N-methylamino)phenyl]ethyl acrylate, and its synthetic route is as follows: [ka]

[0105] This compound is a cyanoacrylate fungicide that acts on Fusarium type I myosin (a motor protein) to cause the fungus to lose its growth motive force and die. ZJS178 has a new structure and unique mode of action, and is not cross-resistant with strobilurin fungicides.

[0106] Preferably, the mass blending ratio of ZJS178 to the strobilurin fungicide is 60:1 to 1:60.

[0107] The strobilurin fungicides include, but are not limited to, azoxystrobin, trifloxystrobin, fluoxastrobin, picoxystrobin, pyraclostrobin, benzenekresoxim-methyl, and phenaminestrobin.

[0108] More preferably, the mass blending ratio of ZJS178 to the strobilurin fungicide is 20:1 to 1:20.

[0109] More preferably, the mass blending ratio of ZJS178 to the strobilurin fungicide is 12:1 to 1:12.

[0110] More preferably, the mass blending ratio of ZJS178 to the strobilurin fungicide is 6:1 to 1:6.

[0111] Most preferably, the mass blending ratio of ZJS178 to the strobilurin fungicide is 3:1 to 1:3.

[0112] The compositions according to the invention are suitable for disinfection, in particular for agricultural disinfection.

[0113] Preferably, the compositions according to the invention are suitable for controlling diseases caused by Fusarium.

[0114] More preferably, the Fusarium refers to plant pathogenic Fusarium, mainly including Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, and Fusarium moniliforme.

[0115] More preferably, the composition according to the present invention is used to control diseases such as cucurbit wilt, tomato wilt, banana wilt, cotton wilt, strawberry wilt, rice Fusarium wilt, and wheat head blight.

[0116] In the disinfecting composition according to the present invention, the disinfectant contains the composition in an amount of 0.5 to 90% by weight.

[0117] More preferably, the disinfectant contains the composition in an amount of 5 to 80% by weight.

[0118] The fungicide according to the present invention may further comprise, in addition to the composition, agriculturally acceptable carriers and adjuvants.

[0119] Because ZJS178 and strobilurin fungicides have different mechanisms of action, the present invention combines the two to significantly improve the control effect against Fusarium disease compared to when the two are used alone, reducing the application rate of each agent, delaying the development of drug resistance in pathogens, and helping to extend the service life of the fungicide.

[0120] Studies have shown that when compound ZJS178 is combined with strobilurin fungicides in certain ratios, it exhibits a synergistic effect in inhibiting the growth of Fusarium. For example, when compound ZJS178 is combined with azoxystrobin in a mass ratio of 3:1 to 1:1, it exhibits a significant synergistic effect in inhibiting the growth of Fusarium oxysporum.

[0121] Furthermore, the fungicide composition further contains auxiliary ingredients necessary for the formulation of an agricultural chemical, and the active ingredient in the composition is 0.5 to 90 mass %. The auxiliary ingredients are carriers and adjuvants commonly used in agricultural chemicals.

[0122] According to methods known to those skilled in the art, the fungicidal compositions described in the present invention can be formulated into various agriculturally acceptable dosage forms, including water dispersible granules, suspensions, and seed treatment suspensions, for practical application.

[0123] For the water dispersible granule formulation, polycarboxylates, lignin sulfonates, alkyl naphthalene sulfonates, etc. can be selected and used as dispersants; alkyl sulfates, alkyl sulfonates, naphthalene sulfonates can be selected and used as wetting agents; ammonium sulfate, urea, sucrose, and glucose can be selected and used as disintegrants; diatomaceous earth, corn starch, polyvinyl alcohol, and carboxymethyl celluloses can be selected and used as binders; and diatomaceous earth, kaolin, white carbon black, light calcium, talc powder, attapulgite, china clay, etc. can be selected and used as fillers.

[0124] The auxiliary agents that can be used in the suspension formulation are as follows: polycarboxylates, lignin sulfonates, and alkylnaphthalene sulfonates are selected as dispersants; alkylphenol polyoxyethylene ether formaldehyde condensate sulfates, alkylphenol polyoxyethylene ether phosphates, phenylethylphenol polyoxyethylene ether phosphates, alkyl sulfates, alkyl sulfonates, and naphthalene sulfonates are selected as wetting agents; xanthan gum, polyvinyl alcohol, and bentonite are selected as thickeners; formaldehyde, benzoic acid, and sodium benzoate are selected as preservatives; silicone-based antifoaming agents are selected as antifreezing agents; and inorganic salts such as ethylene glycol, propylene glycol, glycerin, urea, and sodium chloride are selected as antifreezing agents.

[0125] Adjuvants that can be used in seed treatment suspensions are: Film-forming agents may be polyethylene glycol, polyvinyl alcohol resins, polyvinylpyrrolidone, hydroxyethyl cellulose, homopolyvinyl acetate, sodium carboxymethylcellulose, gum arabic, gelatin, polyvinyl alcohol or polyacrylamide, or mixtures thereof. The technician can adjust the proportion of homopolyvinyl acetate depending on the required viscosity of the product. [Effects of the Invention]

[0126] Compared with the prior art, the present invention has the following beneficial effects.

[0127] (1) The fungicidal composition of the present invention has excellent control effects against Fusarium-caused crop diseases. The combination of ZJS178 with a triazole fungicide exhibits a synergistic effect, significantly improving fungicidal activity and control effects. When ZJS178 is combined with a succinate dehydrogenase inhibitor fungicide, these two components inhibit the mechanical energy (motor proteins) and biological energy (ATP synthesis) of pathogenic fungi, respectively. Although these two components have different mechanisms of action, they are interrelated, and therefore combining the two produces a significant synergistic effect, significantly enhancing fungicidal efficacy. The combination of ZJS178 with a pyrrole fungicide exhibits a synergistic effect, significantly improving fungicidal activity against Fusarium, and significantly improving control effects against seed-borne and soil-borne crop diseases caused by Fusarium. The combination of ZJS178 with thiophanate-methyl or prochloraz exhibits a synergistic effect, significantly improving fungicidal activity and control effects. The combination of compound ZJS178 with strobilurin fungicides has a significant synergistic effect, which can significantly improve the fungicidal activity and control effect.

[0128] (2) The composition according to the present invention is composed of active ingredients with different mechanisms of action, has no cross-resistance, is useful for resolving and delaying drug resistance in pathogens, is safe for crops, and meets the requirements of reducing the amount of pesticide application and improving its efficiency. DETAILED DESCRIPTION OF THE INVENTION

[0129] The present invention will be further described below with reference to specific examples. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and essence of the present invention are included within the scope of the present invention.

[0130] The test methods used in the following examples are conventional methods unless otherwise specified, and the materials and reagents used are commercially available reagents and materials unless otherwise specified. Example 1: Synthesis of compound ZJS178

[0131] Step 1: Synthesis of intermediate b [ka] A reaction flask was charged with 7.0 g of starting material a (CAS number: 67710-36-5) and 150 mL of dichloromethane. 11.7 g of Boc anhydride was added in an ice bath. Next, 6.0 g of triethylamine and 3.3 g of DMAP were slowly added. The mixture was stirred in an ice bath for 0.5 hours, then warmed to room temperature and stirred for an additional 5 hours. After completion of the reaction, 1 M hydrochloric acid was added to the reaction mixture, and the organic phase was separated and washed once with purified water and once with saturated brine. The organic phase was dried over anhydrous Na2SO4 and concentrated to obtain 9.6 g of intermediate b. This crude product was used directly in the next step without further purification.

[0132] Step 2: Synthesis of intermediate c [ka] A reaction flask was charged with 9.6 g of intermediate b, 120 mL of ethanol, and 24 mL of water, and the temperature was raised to 90°C. 3.7 g of iron powder and 12 mL of saturated ammonium chloride solution were added, and the reaction was continued for 4 hours. After the reaction was completed, diatomaceous earth was added and the mixture was filtered. The filtrate was concentrated, and then water was added and extracted three times with ethyl acetate. The organic phases were combined and concentrated, and then separated by column chromatography (eluent: ethyl acetate and petroleum ether, volume ratio: 1:2) to obtain 4.5 g of intermediate c, a yield of 51.1%.

[0133] Step 3: Synthesis of intermediate d [ka] In an ice bath, 0.19 g of sodium hydride was added to 20 mL of tetrahydrofuran, followed by the addition of 1.0 g of intermediate c. The mixture was warmed to room temperature and stirred for 30 minutes. Next, 0.74 g of ethyl iodide was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, water was added to quench the reaction, followed by three extractions with ethyl acetate. The combined organic phases were concentrated and then separated by column chromatography (eluent: ethyl acetate and petroleum ether, volume ratio: 1:5) to obtain 0.52 g of intermediate d, a yield of 48.0%.

[0134] Step 4: Synthesis of intermediate e [ka] In an ice bath, 0.11 g of sodium hydride was added to 10 mL of tetrahydrofuran, followed by the addition of 0.5 g of intermediate d. The mixture was warmed to room temperature and stirred for 30 minutes. Next, 0.40 g of methyl iodide was added, and the mixture was stirred at room temperature overnight. After the reaction was complete, water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and concentrated, and then separated by column chromatography (eluent: ethyl acetate and petroleum ether, volume ratio: 1:5) to obtain 0.34 g of intermediate e, a yield of 65.5%.

[0135] Step 5: Synthesis of compound ZJS178 [ka] A reaction flask was charged with 0.34 g of intermediate e and 10 mL of tetrahydrofuran. Hydrogen chloride gas was produced by adding sodium chloride dropwise to concentrated sulfuric acid. This gas was slowly passed through the reaction mixture. After 1 hour, the gas flow was stopped and the mixture was stirred at room temperature overnight. After the reaction was complete, saturated aqueous sodium bicarbonate was added and the mixture was extracted twice with ethyl acetate. The organic phases were combined and concentrated, then separated by column chromatography (eluent: ethyl acetate and petroleum ether, volume ratio: 1:2) to obtain 0.22 g of compound ZJS178, a yield of 93.2%.

[0136] Example 2: Indoor toxicity measurement of ZJS178 and tebuconazole combination against Fusarium head blight Test subjects: Tebuconazole-susceptible (TS) and resistant (TR) Fusarium graminearum were isolated from the field, and the test strains were identified for morphology, pathogenicity, and resistance, and then stored in the laboratory for future use.

[0137] Test drug: ZJS178 was synthesized in Example 1, and tebuconazole was provided by Zhejiang Chemical Industry Research Institute.

[0138] Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each pesticide was applied at five doses according to its active ingredient content, with the growth inhibition rate for these pathogens ranging from 10% to 90%. The wheat Fusarium head blight pathogen was inoculated onto PDA medium. When colonies grew to cover two-thirds of the Petri dish, a 5mm diameter hole punch was used to create a mycelium mass at the edge of the colony. The mycelium mass was then transferred to the center of a previously prepared pesticide-containing medium plate using an inoculation needle. After incubation in a 25°C incubator for three days, the colony diameter for each treatment was measured with a vernier caliper using the cross method to determine the corrected inhibition rate. Furthermore, the EC50 value for each pesticide was calculated using linear regression analysis between the probability value of the inhibition rate and the logarithm of the concentration series, with four replicates per treatment. ZJS178 was used as the standard pesticide, and the cotoxicity coefficient (CTC) was calculated using the following formula:

[0139] Actual Toxicity Index (ATI) = (EC50 of standard drug / EC50 of test drug) x 100 Theoretical Toxicity Index (TTI) = Toxicity Index of Standard Drug × Percentage of Standard Drug in Mixture + Toxicity Index of Test Drug × Percentage of Test Drug in Mixture Co-toxicity coefficient (CTC) = (ATI / TTI) × 100% When the CTC value was less than 80, the drug combination showed an antagonistic effect, when it was between 80 and 120, it showed an additive effect, and when it was more than 120, it showed a synergistic effect.

[0140] The test results are shown in Tables 1 and 2. [Table 1] [Table 2]

[0141] As can be seen from Tables 1 and 2, ZJS178 had a very strong inhibitory effect against tebuconazole-resistant (TR) and susceptible (TS) wheat head blight fungi, and combinations of ZJS178 with tebuconazole at 2:1, 1:1, and 1:2 ratios showed synergistic effects against both resistant and susceptible strains.

[0142] Example 3: Indoor toxicity measurement of ZJS178 and metconazole combination against rice bakanae disease

[0143] Test subjects: Metconazole-susceptible (MS) and resistant (MR) rice Fusarium fujikuroi were isolated from the field, and the test strains were identified for morphology, pathogenicity, and resistance, and then stored in this laboratory.

[0144] Test drug: ZJS178 was synthesized in Example 1, and metconazole was provided by Zhejiang Chemical Industry Research Institute.

[0145] See Example 2 for bioassay methods.

[0146] The test results are shown in Tables 3 and 4. [Table 3] [Table 4]

[0147] As can be seen from Tables 3 and 4, ZJS178 had a very strong inhibitory effect on the mycelial growth of metconazole-resistant (MR) and susceptible (MS) strains of Gibberella fujikuroi, and combinations of ZJS178 with metconazole at ratios of 3:1, 2:1, 1:1, 1:2, and 1:3 showed synergistic effects against both resistant and susceptible strains.

[0148] Example 4: Indoor toxicity measurement of ZJS178 and tebuconazole combination against Fusarium wilt of strawberry Test subject: Strawberry wilt (Fusarium oxysporum) was isolated from diseased strawberries in the field, and the strain was identified for morphology and pathogenicity and stored in this laboratory. Test drug: ZJS178 was synthesized in Example 1, and tebuconazole was provided by Zhejiang Chemical Industry Research Institute. See Example 2 for bioassay procedures.

[0149] The test results are shown in Table 5. [Table 5]

[0150] As can be seen from the results in Table 5, the combination of ZJS178 and tebuconazole has a synergistic effect and can significantly improve the inhibitory effect of the drug against Fusarium wilt fungus.

[0151] Example 5: Field efficacy test 1. Preparation of Formulations All formulation ratios are expressed as mass percentages. (1) 40% ZJS178 Tebuconazole Water Dispersible Granule Weigh out 10% ZJS178, 30% tebuconazole, 3% TERSPERSE2700, 2% diffusing agent NNO (alkylnaphthalenesulfonate formaldehyde condensate), 3% Nekal BX (sodium dibutylnaphthalenesulfonate), 4% K-12 (sodium lauryl sulfate), 3% diatomaceous earth, and 5% glucose, and add kaolin to make up to 100%. According to the standard method for manufacturing water dispersible granules, the active ingredient, dispersant, wetting agent, binder, etc. were mixed in the appropriate proportions, and then subjected to ultra-fine airflow pulverization. The mixture was then granulated in a granulator equipped with a sieve of a specified size. The mixture was then dried and sieved to obtain a granular product.

[0152] (2) 30% ZJS178 / tebuconazole suspension 15% ZJS178, 15% tebuconazole, 2% TERSPERSE2500, 3% TERSPERSE2425, 0.2% xanthan gum, 3% white carbon black, 5% ethylene glycol, 50.3% benzoic acid, and 0.5% silicone foaming agent were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0153] (3) 25% ZJS178 / Metconazole Suspension 15% ZJS178, 10% metconazole, 2% NNO, 2% TERSPERSE2500, 1% emulsifier T-60, 3% agricultural milk 700#, 0.1% xanthan gum, 3% white carbon black, 5% propylene glycol, 0.5% formaldehyde, and 0.5% silicone antifoaming agent were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0154] (4) 40% ZJS178-tebuconazole microemulsion 25% ZJS178, 15% tebuconazole, 4% TX-10, 6% Agricultural milk 500#, 4% Agricultural milk 1601#, 15% cyclohexanone, 5% N-methylpyrrolidone, 5% n-butanol, and 1% epichlorohydrin were weighed, completely dissolved, and mixed uniformly. Deionized water was added to the mixture up to 100 parts by mass, and the mixture was stirred to prepare a microemulsion. The drug substance, solvent, and emulsifier were added together and dissolved to form a homogeneous oil phase. The water-soluble ingredients were mixed with water to prepare the aqueous phase. The oil and aqueous phases were mixed under high-speed stirring to form a microemulsion.

[0155] (5) 15% ZJS178-tebuconazole water emulsion 10% ZJS178, 5% tebuconazole, 1.5% nonylphenol polyoxyethylene (EO=10) ether phosphate ester, 2.5% triphenylethylphenol polyoxyethylene ether phosphate ester (Agricultural milk 60# phosphate ester), 1% epichlorohydrin, 5% ethylene glycol, 20% xylene, 10% cyclohexanone, 0.3% xanthan gum, and 0.5% benzoic acid were weighed out and added to a total volume of 100 parts by mass with deionized water. The above raw materials were mixed and emulsified under high-speed shear to obtain a water emulsion.

[0156] 2. Field efficacy test for controlling wheat head blight Wheat Fusarium Head Blight control was carried out in accordance with the provisions of "NY / T1464.15-2007 Pesticide Field Efficacy Testing Guidelines - Control of Wheat Fusarium Head Blight with Fungicides." Application was once at the heading and early flowering stages of wheat, and once again six days later. Results were examined at the milk-ripe stage. Samples were taken at five diagonal points for each test plot, with 100-200 ears examined at each point. Grades were assigned based on the percentage of dead ear area relative to the total ear area, and the number of diseased ears and total ears for each grade were recorded. The disease grading criteria were as follows: Grade 0: All spikes are free from disease. Grade 1: The area of dead ears accounts for less than 1 / 4 of the total ear area. Grade 2: The area of dead ears accounts for 1 / 4 to 1 / 2 of the total ear area. Grade 3: The area of dead ears accounts for 1 / 2 to 3 / 4 of the total ear area. Grade 5: The area of dead ears accounts for more than 3 / 4 of the total ear area. Method for calculating efficacy: Based on the survey results, the disease incidence index and chemical control efficacy were calculated using the following formula. Disease index = Σ (number of diseased ears of each grade × relative disease grade) / (total number of ears surveyed × 5) Chemical control effect (%) = (disease index after chemical application in the blank control area - disease index after chemical application in the chemical-treated area) / disease index after chemical application in the blank control area × 100%

[0157] The test results are shown in Table 6. [Table 6] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0158] As can be seen from the results of the above field efficacy tests (Table 6), the combination of ZJS178 and a triazole fungicide effectively controlled wheat head blight, and at the same dose, the combination was significantly superior to the single control fungicides. Within the tested dose range, the combination was safe for the test crop.

[0159] In summary, the results of the laboratory bioassay and field efficacy tests indicate that the composition of the present invention has synergistic effects and exhibits excellent inhibitory and control effects against Fusarium diseases in various crops. This composition, which is composed of active ingredients with different mechanisms of action, is useful in resolving and delaying the development of drug resistance in disease pathogens and is safe for the test crops.

[0160] Example 6: Toxicity determination of the combination of ZJS178 and pydiflumetofen against wheat head blight indoors Test subject: Wheat mold fungus (Fusarium graminearum) isolated and obtained in the field and identified as Fusarium graminearum by biological methods such as morphology and pathogenicity, and stored in this laboratory in preparation for use.

[0161] Test drugs: ZJS178 was synthesized in Example 1, and pydiflumetofen was commercially available from Syngenta Nantong Crop Protection Co., Ltd.

[0162] Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each pesticide was applied at five doses according to its active ingredient content, and the growth inhibition rate for these pathogens ranged from 10% to 90%. The wheat Fusarium head blight pathogen was inoculated onto PDA medium. When colonies grew to cover two-thirds of the Petri dish, a 5mm diameter hole punch was used to create a mycelium mass at the edge of the colony. An inoculation needle was used to transfer the mycelium mass to the center of the previously prepared pesticide-containing medium. After incubation in a 25°C incubator for three days, the colony diameter for each treatment was measured with a vernier caliper using the cross method, and the corrected inhibition rate was calculated. Furthermore, the EC50 value for each pesticide was calculated using linear regression analysis between the probability value of the inhibition rate and the logarithm of the concentration series, with four replicates per treatment. ZJS178 was used as the standard pesticide, and the co-toxicity coefficient (CTC) was calculated using the following formula:

[0163] Actual Toxicity Index (ATI) = (EC50 of standard drug / EC50 of test drug) x 100 Theoretical Toxicity Index (TTI) = Toxicity Index of Standard Drug × Percentage of Standard Drug in Mixture + Toxicity Index of Test Drug × Percentage of Test Drug in Mixture Co-toxicity coefficient (CTC) = (ATI / TTI) × 100% When the CTC value was less than 80, the drug combination showed an antagonistic effect, when it was between 80 and 120, it showed an additive effect, and when it was more than 120, it showed a synergistic effect.

[0164] The test results are shown in Table 7. [Table 7]

[0165] As can be seen from the results in Table 7, both ZJS178 and pydiflumetofen have excellent inhibitory effects against Fusarium head blight, and a combination of the two in a blend ratio range of 5:1 to 1:5 showed a synergistic effect.

[0166] Example 7: Toxicity determination of the combination of ZJS178 and penflufen against rice bakanae disease indoors Test subject: The rice bakanae disease fungus (Fusarium fujikuroi) isolated and obtained in the field and identified as Fusarium fujikuroi by biological methods such as morphology and pathogenicity, and stored in this laboratory.

[0167] Test drugs: ZJS178 was synthesized in Example 1, and penflufen was commercially available from Bayer. See Example 6 for bioassay procedures.

[0168] The test results are shown in Table 8. [Table 8]

[0169] As can be seen from the results in Table 8, both ZJS178 and penflufen had good inhibitory effects against rice seedling disease, and the combination of ZJS178 and penflufen in the range of 5:1 to 1:5 had a significant synergistic effect.

[0170] Example 8: Field efficacy test 1. Preparation of Formulations All formulation ratios are expressed as mass percentages. (1) 40% ZJS178 Pydiflumetofen Water Dispersible Granules Weigh out 20% ZJS178, 20% pydiflumetofen, 3% TERSPERSE 2700, 2% diffusing agent NNO (alkylnaphthalenesulfonate formaldehyde condensate), 3% Nekal BX (sodium dibutylnaphthalenesulfonate), 4% K-12 (sodium lauryl sulfate), 3% diatomaceous earth, and 5% glucose, and add kaolin to make up to 100%. According to the standard method for manufacturing water dispersible granules, the active ingredient, dispersant, wetting agent, binder, etc. were mixed in the appropriate proportions, and then subjected to ultra-fine airflow pulverization. The mixture was then granulated in a granulator equipped with a sieve of a specified size. The mixture was then dried and sieved to obtain a granular product.

[0171] (2) 10% ZJS178-Penflufen Suspension 5% ZJS178, 5% penflufen, 2% TERSPERSE 2500, 3% TERSPERSE 2425, 0.2% xanthan gum, 3% white carbon black, 5% ethylene glycol, 0.5% formaldehyde, 0.3% benzoic acid, and 0.5% silicone antifoaming agent (trade name: s-29, manufactured by Nanjing Sixin Applied Chemicals Co., Ltd.) were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0172] 2. Field Test 2.1 Field efficacy test for wheat Fusarium head blight control Wheat Fusarium Head Blight control was carried out in accordance with the provisions of "NY / T1464.15-2007 Pesticide Field Efficacy Testing Guidelines - Control of Wheat Fusarium Head Blight with Fungicides." Application was once at the heading and early flowering stages of wheat, and once again six days later. Two weeks after application, inspections were conducted. Samples were taken at five diagonal points for each test plot, with 100-200 spikelets inspected at each point. Grades were assigned based on the percentage of dead spikelet area relative to the total spikelet area, and the number of diseased spikelets and total spikelets for each grade were recorded. The disease grading criteria were as follows: Grade 0: All spikes are free from disease. Grade 1: The area of dead ears accounts for less than 1 / 4 of the total ear area. Grade 2: The area of dead ears accounts for 1 / 4 to 1 / 2 of the total ear area. Grade 3: The area of dead ears accounts for 1 / 2 to 3 / 4 of the total ear area. Grade 4: The area of dead ears accounts for more than 3 / 4 of the total ear area. Disease index = Σ (number of diseased ears of each grade × relative disease grade) / (total number of ears surveyed × 5) Chemical control effect (%) = (disease index after chemical application in the blank control area - disease index after chemical application in the chemical-treated area) / disease index after chemical application in the blank control area × 100%

[0173] The test results are shown in Table 9. [Table 9] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0174] As can be seen from the results in Table 9, the combination of ZJS178 and pydiflumetofen has excellent control effect against wheat head blight. Compared with the single agents, the combination of the two at the same dose has a significant synergistic effect.

[0175] 2.2 Field efficacy test for the control of rice bakanae disease The control of rice bakanae disease was carried out in accordance with the provisions of GB-T 17980.104-2004, "Guidelines for Field Efficacy Tests of Pesticides (II) - Control of Rice Bakanae Disease with Fungicides." The test site was located in Dongpu Town, Shaoxing City, and the rice variety used was Zhongzao 39 (a susceptible variety). The chemical solution was diluted to a certain ratio, and seeds were soaked for 72 hours, germinated, and then sown. Seedlings were grown in nutrient soil in strict accordance with the mechanical transplant seedling cultivation process. Seed volume and soil fertility were consistent across treatments. Twenty seedling trays were treated each time, with 150g of seeds per tray. The incidence of bakanae disease was monitored one day before rice transplanting and before heading. The rate of diseased plants and control efficacy were calculated using the following formula: Diseased plant rate (%) = Number of diseased plants / Number of plants surveyed x 100 Chemical control effect (%) = (disease rate in blank control plot - disease rate in chemical-treated plot) / disease rate in blank control plot x 100

[0176] The test results are shown in Table 10. [Table 10] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0177] As can be seen from the results of the field efficacy test in Table 10, the combination of ZJS178 and penflufen has excellent control effect against rice seedling disease. Under the same effective dose, the control effect of the combination is significantly better than that of either agent alone. It was safe for the test crops within the test dose range.

[0178] In summary, as can be seen from the results of the indoor bioassay and the field efficacy test, the fungicide composition of the present invention has a significant synergistic effect, and the composition is composed of active ingredients with different mechanisms of action, which can delay the development of drug resistance in pathogens against a single drug. The combination did not have any obvious adverse effects on the test crops, and the leaf color and growth were all normal.

[0179] Example 9: Determining the toxicity of ZJS178 and fludioxonil in combination against Fusarium pseudograminalum on wheat in the laboratory Test subjects: Fludioxonil-resistant (FR) and susceptible (FS) Fusarium pseudograminearum on wheat, which were field isolated, identified, and stored by the laboratory. Test drug: ZJS178 was synthesized in Example 1, and fludioxonil was provided by Zhejiang Chemical Industry Research Institute. Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each pesticide was administered at five doses according to the active ingredient content. The above-mentioned wheat Fusarium pseudograminalum was inoculated onto PDA medium. When colonies grew to cover two-thirds of the Petri dish, a 5 mm diameter hole punch was used to create a mycelium mass at the edge of the colony. The mycelium mass was then transferred to the center of the previously prepared pesticide-containing PDA medium using an inoculation needle. After incubation in a 25°C incubator for three days, the colony diameter (cm) for each treatment was measured with a vernier caliper using the cross method, and the corrected inhibition rate was calculated. Furthermore, the EC50 value of each pesticide was calculated using linear regression analysis between the probability value of the inhibition rate and the logarithm of the serial concentrations. Four replicates were performed for each treatment. ZJS178 was used as the standard pesticide, and the co-toxicity coefficient (CTC) was calculated using the following formula: Actual Toxicity Index (ATI) = (EC50 of standard drug / EC50 of test drug) x 100 Theoretical Toxicity Index (TTI) = Toxicity Index of Standard Drug × Percentage of Standard Drug in Mixture + Toxicity Index of Test Drug × Percentage of Test Drug in Mixture Co-toxicity coefficient (CTC) = (ATI / TTI) × 100% When the CTC value was less than 80, the drug combination showed an antagonistic effect, when it was between 80 and 120, it showed an additive effect, and when it was more than 120, it showed a synergistic effect.

[0180] The test results are shown in Tables 11 and 12. [Table 11] [Table 12]

[0181] As can be seen from the results in Tables 11 and 12, ZJS178 has good inhibitory effects against both fludioxonil-resistant (FR) and susceptible (FS) Fusarium pseudograminalum. The rational combination of ZJS178 and fludioxonil produced additive or synergistic effects against resistant and susceptible strains, but no antagonistic effects.

[0182] Example 10: Indoor toxicity measurement of the combination of ZJS178 and fludioxonil against Gibberella f.sp. Test subject: Fusarium fujikuroi, a rice seedling disease fungus kept in the laboratory's fungal seedling room. Test drug: ZJS178 was synthesized in Example 1, and fludioxonil was provided by Zhejiang Chemical Industry Research Institute. See Example 9 for bioassay procedures.

[0183] The test results are shown in Table 13. [Table 13]

[0184] As can be seen from the results in Table 13, both ZJS178 and fludioxonil had excellent inhibitory effects against rice Bakanae disease, and the combination of the two showed additive or synergistic effects.

[0185] Example 11: Toxicity determination of ZJS178 and fludioxonil combination against tomato wilt indoors Test subject: Fusarium oxysporum, the pathogen of tomato wilt, stored in the laboratory's seed room. Test drug: ZJS178 was synthesized in Example 1, and fludioxonil was provided by Zhejiang Chemical Industry Research Institute. See Example 9 for bioassay methods.

[0186] The test results are shown in Table 14. [Table 14]

[0187] As can be seen from the results in Table 14, both ZJS178 and fludioxonil had significant inhibitory effects against Fusarium wilt of tomato, and the rational combination of the two showed additive or synergistic effects.

[0188] Example 12: Field efficacy test Preparation of 1.25% ZJS178·fludioxonil suspension All percentages in the formulation ratios of the seed treatment suspension are by mass. The active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the following proportions using water as the medium, mixed uniformly, dispersed in a ball mill or high-speed shearing machine for 30 minutes, and then pulverized in a sand mill to prepare the suspension. 15% ZJS178, 10% fludioxonil, 2% TERSPERSE 2500, 3% TERSPERSE 2425, 0.2% xanthan gum, 3% white carbon black, 5% ethylene glycol, 0.3% benzoic acid, and 0.5% silicone foaming agent (trade name: s-29, manufactured by Nanjing Sixin Applied Chemicals Co., Ltd.) were weighed out and added with deionized water up to 100 parts by mass.

[0189] 2. Field Test Field application example 1: Field efficacy test for the control of rice bakanae disease The control of rice bakanae disease was carried out in accordance with the provisions of GB-T 17980.104-2004, "Guidelines for Field Efficacy Tests of Pesticides (II) - Control of Rice Bakanae Disease with Fungicides." The test site was located in Dongpu Town, Shaoxing City, and the rice variety used was Zhongzao 39 (a susceptible variety). The chemical solution was diluted to a specific ratio, and seeds were soaked for 72 hours, germinated, and then sown. Seedlings were grown in nutrient soil in strict accordance with the mechanical transplantation seedling cultivation process. The seed amount and soil fertility were consistent across treatments. Twenty seedling trays were treated each time, with 150g of seeds per tray. The incidence of bakanae disease was monitored one day before rice transplanting and before heading. The rate of diseased plants and control efficacy were calculated using the following formula: Diseased plant rate (%) = Number of diseased plants / Number of plants surveyed x 100 Chemical control effect (%) = (disease rate in blank control plot - disease rate in chemical-treated plot) / disease rate in blank control plot x 100

[0190] The test results are shown in Table 15. [Table 15] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0191] As can be seen from the results in Table 15, the control effect of the combination of ZJS178 and fludioxonil against rice seedling disease was significantly superior to that of either agent alone under the same effective dose and seed soaking conditions.

[0192] Field application example 2. Field efficacy test for controlling wheat root rot The test site was located in Dongpu Town, Shaoxing City. The soil type was loam, with moderate fertility and a neutral pH. The wheat variety was Jimai 22. The seed volume and soil fertility were consistent across treatments. The seeds were coated with a fixed dilution of the chemical solution, with 2% of the seed volume used for water coating. After coating, the seeds were placed in a well-ventilated, dark place to dry. Three plots were set up for each treatment. Samples were taken at five points in each plot before heading. 100 plants were examined at each point, and the total number of plants and the number of diseased plants were recorded. Based on the results, the diseased plant rate and control efficacy were calculated using the following formula. The test data were statistically analyzed using Duncan's multiple range test (DMRT). Diseased plant rate (%) = Number of diseased plants / Number of plants surveyed x 100 Chemical control effect (%) = (disease rate in blank control plot - disease rate in chemical-treated plot) / disease rate in blank control plot x 100

[0193] The test results are shown in Table 16. [Table 16] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0194] As can be seen from the results in Table 16, under the same effective dose and seed dressing conditions, the control effect of the combination of ZJS178 and fludioxonil against wheat root rot was significantly superior to that of each single agent.

[0195] To summarize the above, as can be seen from the results of indoor bioassays and field efficacy tests, the combination of the composition of the present invention has a significant synergistic effect, and the composition is composed of active ingredients with different mechanisms of action, which helps to resolve and delay the occurrence of drug resistance in pathogens. Furthermore, there was no obvious adverse effect on the test crops, and the leaf color and growth were all normal, making it safe.

[0196] Example 13: Toxicity determination of the combination of ZJS178 and thiophanate methyl against watermelon wilt indoors Test subject: Watermelon wilt fungus (F. Oxysporum f.sp. niveum) was isolated in the field, and the test strain was identified for morphology and pathogenicity, and then stored in this laboratory for use. Test drug: ZJS178 was synthesized in Example 1, and thiophanate methyl was provided by Zhejiang Chemical Industry Research Institute. Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each pesticide was applied at five doses according to its active ingredient content, with the growth inhibition rate of these pathogens ranging from 10% to 90%. The watermelon wilt fungus was inoculated onto PDA medium. When colonies grew to cover two-thirds of the Petri dish, a 5mm diameter hole punch was used to create a mycelium mass at the edge of the colony. The mycelium mass was then transferred to the center of a previously prepared pesticide-containing medium plate using an inoculation needle. After incubation in a 25°C incubator for three days, the colony diameter for each treatment was measured with a vernier caliper using the cross method to determine the corrected inhibition rate. Furthermore, the EC50 value of each pesticide was calculated using linear regression analysis between the probability value of the inhibition rate and the logarithm of the concentration series, with four replicates per treatment. ZJS178 was used as the standard pesticide, and the co-toxicity coefficient (CTC) was calculated using the following formula: Actual Toxicity Index (ATI) = (EC50 of standard drug / EC50 of test drug) x 100 Theoretical Toxicity Index (TTI) = Toxicity Index of Standard Drug × Percentage of Standard Drug in Mixture + Toxicity Index of Test Drug × Percentage of Test Drug in Mixture Co-toxicity coefficient (CTC) = (ATI / TTI) × 100% When the CTC value was less than 80, the drug combination showed an antagonistic effect, when it was between 80 and 120, it showed an additive effect, and when it was more than 120, it showed a synergistic effect.

[0197] The test results are shown in Table 17. [Table 17]

[0198] As shown in Table 17, when the mass percentage of ZJS178 and thiophanate methyl was 6:1 to 1:3, there was a significant synergistic inhibitory effect on the growth of watermelon wilt fungus, and there was an additive effect at other blending ratios.

[0199] Example 14: Determining the toxicity of ZJS178 and prochloraz in combination against wheat head blight indoors Test subject: Wheat head blight (Fusarium graminearum) was kept in the strain room of this laboratory. Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each agent was treated at five doses based on the active ingredient content. The wheat Fusarium head blight pathogen was inoculated onto PDA medium. When colonies grew to cover two-thirds of the Petri dish, a 5mm diameter hole punch was used to create a mycelium mass at the edge of the colony. The mycelium mass was then transferred to the center of pre-prepared toxicant-containing PDA medium using an inoculation needle. The plates were then incubated in an incubator at 25°C. Four replicates were performed for each treatment. Based on the CK colony growth, the colony diameter (cm) for each treatment was measured with a vernier caliper using the cross method to determine the corrected inhibition rate. The EC50 value for each agent was calculated using linear regression analysis between the probability of inhibition and the logarithm of the concentration series, and the co-toxicity coefficient (CTC) was calculated using the Sun Yunpei method. Calculation of efficacy: Two diameters of each colony were measured using the cross method, and the average was used to represent the colony size. The colony growth inhibition rate was calculated using the formula: % colony growth inhibition = (increased diameter of blank control colony - increased diameter of drug-treated colony) x 100 / increased diameter of blank control colony.

[0200] [Table 18]

[0201] As shown in Table 18, when the mass percentage of ZJS178 and prochloraz was 1:15 to 12:1, there was a significant synergistic inhibitory effect on the growth of Fusarium head blight fungus, while there was an additive effect at other mixing ratios.

[0202] Example 15: Determining the Toxicity of ZS178 in Combination with Prochloraz-Manganese Chloride Complex against Strawberry Wilt Indoors Test subject: Strawberry wilt (Fusarium oxysporum) was kept in the strain room of this laboratory. For bioassay method, see Example 14.

[0203] [Table 19]

[0204] As shown in Table 19, when the mass percentage of ZJS178 and prochloraz-manganese chloride complex was 1:12 to 15:1, there was a significant synergistic inhibitory effect on the growth of strawberry wilt fungus, and there was an additive effect at other combination ratios.

[0205] Example 16: Field efficacy test 1. Preparation of Formulations All formulation ratios are expressed as mass percentages. (1) 20% ZJS178 thiophanate methyl suspension 10% ZJS178, 10% thiophanate methyl, 2% TERSPERSE 2500, 3% TERSPERSE 2425, 0.2% xanthan gum, 3% white carbon black, 5% ethylene glycol, 0.3% benzoic acid, and 0.5% silicone antifoaming agent were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0206] (2) 46% ZJS178 Prochloraz Suspension 23% ZJS178, 23% prochloraz, 2% NNO, 2% TERSPERSE 2500, 1% emulsifier T-60, 3% agricultural milk 700#, 0.1% xanthan gum, 3% white carbon black, 5% propylene glycol, 0.5% formaldehyde, and 0.5% silicone antifoaming agent were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0207] 2. Field Test 2.1 Field efficacy test for wheat Fusarium head blight control Wheat Fusarium Head Blight control was carried out in accordance with the provisions of "NY / T1464.15-2007 Pesticide Field Efficacy Testing Guidelines - Control of Wheat Fusarium Head Blight with Fungicides." A single application was performed at the heading and early flowering stages of wheat, with results being assessed at the milk-ripe stage. Samples were taken at five diagonal points for each test plot, with 100-200 ears examined at each point. Grades were assigned based on the percentage of dead ear area relative to the total ear area, and the number of diseased ears and total ears for each grade were recorded. The disease grading criteria were as follows: Grade 0: All spikes are free from disease. Grade 1: The area of dead ears accounts for less than 1 / 4 of the total ear area. Grade 3: The area of dead ears accounts for 1 / 4 to 1 / 2 of the total ear area. Grade 5: The area of dead ears accounts for 1 / 2 to 3 / 4 of the total ear area. Grade 7: The area of dead ears accounts for more than 3 / 4 of the total ear area. Calculation method of efficacy: Based on the survey results, the disease index and chemical control efficacy were calculated using the following formulas (1) and (2). The test data were statistically analyzed using Duncan's new multiple range test (DMRT). Disease index = Σ [number of diseased spikelets for each grade × relative grade value] / (total number of surveys × 7) × 100 (1) Control effect (%) = (1-(PT1 / CK1)) x 100 (2) In the formula, CK1 is the disease index after application of the drug to the blank control plot, and PT1 is the disease index after application of the drug to the drug-treated plot.

[0208] [Table 20] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0209] As can be seen from the results in Table 20, 46% of ZJS178 / prochloraz suspension had good control effects against wheat head blight at doses of 5g / mu to 15g / mu of active ingredient, with control rates ranging from 91.31% to 97.88%, superior to the control single agent. The test agent was safe for the test crops.

[0210] 2.2 Field efficacy test for tomato wilt control The test site was located in Dongpu Town, Shaoxing City. The soil type was loam, with moderate fertility and a neutral pH. The tomato variety was Zhefun 202. After transplanting and planting, the tomato seedlings were treated by root irrigation with a chemical solution. The type and application rate of the chemical are shown in Table 5. One week after the initial application, root irrigation was repeated. 21 days after treatment, the incidence of tomato wilt was investigated. Samples were taken at five random points for each treatment, covering an area of 5 square meters. The total number of tomato plants and the number of diseased plants were counted at each point. The diseased plant rate and control efficacy were calculated. Control effect (%) = ((control disease rate - treatment disease rate) / control disease rate) x 100

[0211] The test results are shown in Table 21. [Table 21] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0212] As can be seen from the results in Table 21, when the effective doses were the same, the combination of ZJS178 and thiophanate-methyl effectively controlled tomato wilt, and the control effect was significantly better than that of either agent alone. The test agents were safe for the test crops.

[0213] In summary, the results of the laboratory bioassay and field efficacy tests show that the composition of the present invention has rational ingredients, has therapeutic and protective effects, excellent fungicidal effect, reduces the number of applications, and is low in application cost. Furthermore, its activity and fungicidal effect are not simply the sum of the activities of the individual components but have significant synergistic effects. The composition is composed of active ingredients with different mechanisms of action, which increases the number of sites of action, thereby helping to prevent and delay the development of drug resistance in pathogens. Furthermore, the composition had no obvious adverse effects on the test crops, and leaf color and growth were all normal. It is safe and meets the safety requirements for pesticide formulations. The present invention has excellent control effects against Fusarium diseases.

[0214] Example 17: Toxicity determination of the combination of ZJS178 and azoxystrobin against tomato wilt indoors Test subject: Tomato wilt fungus (F. Oxysporum f.sp. lycopersici) was isolated from diseased tomatoes, and the test strain was identified for morphology and pathogenicity, and then stored in this laboratory for use. Test drug: ZJS178 was synthesized in Example 1, and azoxystrobin was provided by Zhejiang Chemical Industry Research Institute. Test method: The mycelium growth rate method of the "People's Republic of China Agricultural Industry Standard NY / T 1156.2-2006" was used. Each pesticide was applied at five doses according to its active ingredient content, with the growth inhibition rate of these pathogens ranging from 10% to 90%. The tomato wilt fungus was inoculated onto PDA medium. To inhibit bypass respiration, 50 μg / mL salicylhydroxamic acid (SHAM) was added to the medium. When colonies grew to cover two-thirds of the Petri dish, a 5 mm diameter hole punch was used to create a mycelium mass at the edge of the colony. An inoculation needle was used to transfer the mycelium mass to the center of a previously prepared pesticide-containing medium plate. After incubation in a 25°C incubator for three days, the colony diameter for each treatment was measured with a vernier caliper using the cross method to determine the corrected inhibition rate. Furthermore, the EC50 value for each pesticide was calculated using linear regression analysis between the probability of inhibition and the logarithm of the concentration series. Four replicates were performed for each treatment. ZJS178 was set as the standard drug, and the co-toxicity coefficient (CTC) was calculated using the following formula. Actual Toxicity Index (ATI) = (EC50 of standard drug / EC50 of test drug) x 100 Theoretical Toxicity Index (TTI) = Toxicity Index of Standard Drug × Percentage of Standard Drug in Mixture + Toxicity Index of Test Drug × Percentage of Test Drug in Mixture Co-toxicity coefficient (CTC) = (ATI / TTI) × 100% When the CTC value was less than 80, the drug combination showed an antagonistic effect, when it was between 80 and 120, it showed an additive effect, and when it was more than 120, it showed a synergistic effect.

[0215] The test results are shown in Table 22. [Table 22]

[0216] As shown in Table 22, when the mass percentage of ZJS178 and azoxystrobin was 4:1 to 1:6, there was a significant synergistic inhibitory effect on the growth of tomato wilt fungus, and there was an additive effect at other blending ratios.

[0217] Example 18: Toxicity measurement of the combination of ZJS178 and pyraclostrobin against rice bakanae disease indoors Test subject: Fusarium moniliforme, a bacillus of rice caused by the bacillus subtilis of the rice plant, was kept in the strain room of this laboratory. For the bioassay method, see Example 17.

[0218] [Table 23]

[0219] As shown in Table 23, when the mass percentage of ZJS178 and pyraclostrobin was 1:12 to 8:1, there was a significant synergistic inhibitory effect on the growth of the rice pseudomonas fuscata, while there was an additive effect at other combination ratios.

[0220] Example 19: Field efficacy test 1. Preparation of Formulations All formulation ratios are expressed as mass percentages. (1) 40% ZJS178 Azoxystrobin Suspension 30% ZJS178, 10% azoxystrobin, 2% NNO, 2% TERSPERSE 2500, 1% emulsifier T-60, 3% agricultural milk 700#, 0.1% xanthan gum, 3% white carbon black, 5% propylene glycol, 0.5% formaldehyde, and 0.5% silicone antifoaming agent were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0221] (2) 24% ZJS178-pyraclostrobin suspension 16% ZJS178, 8% pyraclostrobin, 2% TERSPERSE 2500, 3% TERSPERSE 2425, 0.2% xanthan gum, 3% white carbon black, 5% ethylene glycol, 0.3% benzoic acid, and 0.5% silicone antifoaming agent (trade name: s-29, manufactured by Nanjing SIXIN Applied Chemicals Co., Ltd.) were weighed out and added with deionized water up to 100 parts by mass. Using water as a medium, the active ingredient, dispersant, suspending agent, antifreeze agent, etc. were added to a blending kettle in the appropriate proportions and mixed uniformly, then dispersed in a ball mill or high-speed shearing machine for 30 minutes, and pulverized in a sand mill to prepare a suspension.

[0222] 2. Field Test Field efficacy trials for the control of watermelon wilt Watermelon wilt control was conducted in accordance with GB-T 17980.113-2004 Guidelines for Field Efficacy Tests of Pesticides (II) - Control of Watermelon Wilt with Fungicides. The test site was located in Dongpu Town, Shaoxing City. The soil type was loam, with moderate fertility and a neutral pH. The watermelon cultivar was Zhemi No. 8. After transplanting and planting, watermelon seedlings were treated with a root irrigation solution. Control plants were sprayed with a fungicide. The fungicide types and application rates are listed in Table 3. One week after the initial application, root irrigation was repeated. 14 days after treatment, all plants in the plot were inspected for typical wilt symptoms. The percentage of infected plants and the total number of plants inspected were recorded. Based on the results of the investigation, the percentage of infected plants and control efficacy were calculated using the following formulas (1) and (2). The test data were statistically analyzed using Duncan's new multiple range test (DMRT). Disease incidence rate (%) = Number of diseased plants / Number of surveyed plants × 100 (1) Control effect (%) = (rate of diseased plants in blank control plots - rate of diseased plants in pesticide-treated plots) / rate of diseased plants in blank control plots × 100 (2)

[0223] The test results are shown in Table 24. [Table 24] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0224] As can be seen from the results in Table 24, when the effective doses were the same, the combination of ZJS178 and azoxystrobin had excellent control effect against watermelon wilt, and the control effect was significantly higher than that of either agent alone. The test pesticides were safe for the test crops.

[0225] Field efficacy test for the control of rice bakanae disease The control of rice bakanae disease was carried out in accordance with the provisions of GB-T 17980.104-2004, "Guidelines for Field Efficacy Tests of Pesticides (II) - Control of Rice Bakanae Disease with Fungicides." Samples were taken at five points per plot, with 100 plants examined at each point. The total number of plants and the number of diseased plants were recorded. In the field, samples were taken at five random points per plot before heading, with 20 groups collected at each point. The test site was located in Dongpu Town, Shaoxing City. The chemical solution was diluted to a certain ratio and seeds were soaked for 72 hours. The soil at the test site was loamy, medium fertility, and neutral pH. The rice variety used was Chu-Hai 39 (a susceptible variety). Seedlings were grown using nutrient soil in strict accordance with the mechanical transplant seedling cultivation process. Seed volume and soil fertility were consistent across treatments. Twenty seedling trays were treated each time, with 150g of seeds per tray. The incidence of Bakanae disease was investigated one day before rice transplanting and before heading. The efficacy was calculated as follows: Based on the survey results, the disease index and control effect were calculated using the following formulas (1) and (2): The test data were statistically analyzed using Duncan's new multiple range test (DMRT). Disease incidence rate (%) = Number of diseased plants / Number of surveyed plants × 100 (1) Control effect (%) = (rate of diseased plants in blank control plots - rate of diseased plants in pesticide-treated plots) / rate of diseased plants in blank control plots × 100 (2)

[0226] [Table 25] *Different lowercase letters following data in the same column indicate significant differences at the P<0.05 level.

[0227] As can be seen from the results in Table 25, soaking seeds in a 24% ZJS178 / pyraclostrobin suspension diluted 3000-2000 times had a good effect on rice seedling disease, with control rates reaching 89.8%-96.2%, significantly better than that of the single agent. The test agents were safe for the test crops.

[0228] In summary, the results of the laboratory bioassay and field efficacy tests show that the composition of the present invention has rational ingredients, has therapeutic and protective effects, excellent fungicidal effect, reduces the number of applications, and is low in application cost. Furthermore, its activity and fungicidal effect are not simply the sum of the activities of the individual components but have significant synergistic effects. The composition is composed of active ingredients with different mechanisms of action, which increases the number of sites of action, thereby helping to prevent and delay the development of drug resistance in pathogens. Furthermore, the composition had no obvious adverse effects on the test crops, and leaf color and growth were all normal. It is safe and meets the safety requirements for pesticide formulations. The present invention has excellent control effects against Fusarium diseases.

[0229] The present invention has excellent control effects against seed-borne and soil-borne crop diseases caused by Fusarium.

Claims

1. A fungicidal composition characterized in that the active ingredients comprise a compound ZJS178 having the structural formula shown in formula (I) and a triazole fungicide. 【Chemical 1】

2. 2. The fungicidal composition according to claim 1, wherein the triazole fungicide is tebuconazole or metconazole.

3. 2. The fungicidal composition according to claim 1, wherein the weight ratio of compound ZJS178 to triazole fungicide in the composition is 50:1 to 1:

50.

4. 4. The fungicidal composition according to claim 3, wherein the mass ratio of compound ZJS178 to triazole fungicide in the composition is 8:1 to 1:

8.

5. The fungicidal composition according to claim 4, wherein the mass ratio of compound ZJS178 to triazole fungicide in the composition is 2:1 to 1:

3.

6. 6. The fungicidal composition according to claim 1, further comprising auxiliary ingredients necessary for the formulation of a pesticide, and the active ingredient in the composition is 1 to 90% by mass.

7. 7. The fungicidal composition according to claim 6, wherein the mass percentage of the active ingredient is 15 to 40%.

8. Use of the fungicidal composition according to any one of claims 1 to 7 in controlling crop diseases caused by Fusarium.

9. The use according to claim 8, characterized in that the Fusarium is Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, or Fusarium moniliforme.

10. The use according to claim 9, characterized in that the crop diseases include wheat head blight, rice seedling disease, and strawberry wilt.

11. An efficient fungicidal composition, characterized in that the active ingredients comprise a compound ZJS178 having the structural formula shown in formula (I) and a succinate dehydrogenase inhibitor fungicide. 【Chemistry 2】

12. The efficient fungicidal composition according to claim 11, characterized in that the mass ratio of compound ZJS178 to succinate dehydrogenase inhibitor fungicide in said composition is 50:1 to 1:

50.

13. 12. The efficient disinfectant composition according to claim 11, wherein the succinate dehydrogenase inhibitor disinfectant is pydiflumetofen or penflufen.

14. 14. An efficient fungicidal composition according to claim 12 or 13, characterized in that the mass ratio of compound ZJS178 to succinate dehydrogenase inhibitor fungicide in said composition is 9:1 to 1:

9.

15. 15. The efficient fungicidal composition according to claim 14, characterized in that the mass ratio of compound ZJS178 to succinate dehydrogenase inhibitor fungicide in said composition is preferably 7:1 to 1:

5.

16. An efficient disinfecting composition according to claim 11, characterized in that the active ingredient in the composition is 10 to 40% by weight.

17. 12. The efficient fungicidal composition of claim 11, wherein the composition further comprises an agriculturally acceptable carrier and an adjuvant.

18. Use of an efficient fungicidal composition according to any one of claims 11 to 17 in controlling Fusarium diseases in crops.

19. The use according to claim 18, characterized in that the Fusarium is Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, or Fusarium moniliforme.

20. The use according to claim 19, characterized in that the diseases include wheat head blight and rice seedling disease.

21. A fungicide composition comprising as active ingredients a compound ZJS178 having the structural formula represented by formula (I) and a pyrrole fungicide. 【Chemistry 3】

22. 22. The fungicide composition of claim 21, wherein the pyrrole fungicide is fludioxonil.

23. 23. The fungicide composition according to claim 21 or 22, wherein the mass ratio of the compound ZJS178 to the pyrrole fungicide is 50:1 to 1:

50.

24. 24. The fungicide composition according to claim 23, wherein the mass ratio of compound ZJS178 to pyrrole fungicide is 11:1 to 1:

11.

25. The fungicide composition according to claim 24, wherein the mass ratio of the compound ZJS178 to the pyrrole fungicide is 1:1-3.

26. 22. The fungicide composition according to claim 21, wherein the active ingredient in the composition is 25 to 40% by weight.

27. 22. The fungicide composition of claim 21 further comprising pesticidally acceptable carriers and adjuvants.

28. Use of the fungicide composition according to any one of claims 21 to 27 in controlling crop diseases caused by Fusarium.

29. The use according to claim 28, characterized in that the Fusarium is Fusarium pseudograminalum, Fusarium fujikuroi, or Fusarium oxysporum.

30. 30. The use according to claim 29, comprising preparing the fungicide composition into a seed treatment to treat seeds of crops.

31. A fungicide composition, characterized in that the active ingredients comprise a compound ZJS178 having a structure represented by formula (I) and an imidazole fungicide. 【Chemistry 4】

32. The fungicide composition according to claim 31, wherein the imidazole fungicide is at least one selected from prochloraz, carbendazim, prochloraz-manganese chloride complex, and thiophanate methyl.

33. The fungicide composition according to claim 31, wherein the mass ratio of ZJS178 to the imidazole fungicide in the composition is 60:1 to 1:

60.

34. The fungicide composition according to claim 33, wherein the mass ratio of ZJS178 to the imidazole fungicide in the composition is 20:1 to 1:

20.

35. The fungicide composition according to claim 34, wherein the mass ratio of ZJS178 to the imidazole fungicide in the composition is 12:1 to 1:

12.

36. The fungicide composition according to claim 35, wherein the mass ratio of ZJS178 to the imidazole fungicide in the composition is 6:1 to 1:

6.

37. The fungicide composition according to claim 36, wherein the mass ratio of ZJS178 to the imidazole fungicide in the composition is 3:1 to 1:

3.

38. The fungicide composition according to any one of claims 31 to 37, further comprising auxiliary ingredients necessary for the formulation of an agricultural chemical, and the active ingredient in the composition is 0.5 to 90% by mass.

39. Use of the fungicide composition according to any one of claims 31 to 37 in controlling crop diseases caused by Fusarium.

40. 40. The use according to claim 39, wherein the Fusarium is Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, or Fusarium moniliforme.

41. 41. The use according to claim 40, wherein the crop diseases include cucurbit wilt, tomato wilt, banana wilt, cotton wilt, strawberry wilt, rice seedling disease, and wheat head blight.

42. A fungicidal composition characterized in that the active ingredients comprise a compound ZJS178 having the structure shown in formula (I) and a strobilurin fungicide. 【Chemistry 5】

43. 43. The fungicidal composition of claim 42, wherein the strobilurin fungicide is at least one selected from azoxystrobin, trifloxystrobin, fluoxastrobin, picoxystrobin, pyraclostrobin, and phenaminestrobin.

44. 43. The fungicidal composition according to claim 42, wherein the mass ratio of ZJS178 to the strobilurin fungicide in the composition is 60:1 to 1:

60.

45. 45. The fungicidal composition according to claim 44, wherein the mass ratio of ZJS178 to the strobilurin fungicide in the composition is 20:1 to 1:

20.

46. 46. The fungicidal composition according to claim 45, wherein the mass ratio of ZJS178 to the strobilurin fungicide in the composition is 12:1 to 1:

12.

47. 47. The fungicidal composition according to claim 46, wherein the mass ratio of ZJS178 to the strobilurin fungicide in the composition is 6:1 to 1:

6.

48. 48. The fungicidal composition according to claim 47, wherein the mass ratio of ZJS178 to the strobilurin fungicide in the composition is 3:1 to 1:

3.

49. The fungicidal composition according to any one of claims 42 to 48, further comprising auxiliary ingredients necessary for the formulation of a pesticide, and the active ingredient in the composition is 0.5 to 90% by mass.

50. Use of the fungicidal composition according to any one of claims 42 to 48 in controlling crop diseases caused by Fusarium.

51. The use according to claim 50, characterized in that the Fusarium is Fusarium graminearum species complex, Fusarium fujikuroi species complex, Fusarium oxysporum, or Fusarium moniliforme.

52. 52. The use according to claim 51, wherein the crop diseases include cucurbit wilt, tomato wilt, banana wilt, cotton wilt, strawberry wilt, rice seedling disease, and wheat head blight.

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