Method for controlling or preventing infestation of useful plants by phytopathogenic microorganisms

Abstract

The present invention relates to a method for controlling or preventing an infestation of useful plants by a phytopathogenic microorganism selected from the group consisting of Fusarium pseudograminearum, Fusarium culmorum, Fusarium oxysporum, Fusarium graminearum, Bakania oryzae, Sporidesmium humicola, Botrytis cinerea, Phytophthora sojae, Phytophthora capsici, Phytophthora infestans, Pythium aphanidermatum, Pythium arrhenomanes, Pythium deliense, Puccinia striiformis, Puccinia hordei, Puccinia malvaccarum, Pseudoperonospora cubensis and Sphaerotheca fuliginea, which comprises the application of a compound of formula (I) to useful plants, to the locus thereof, or to their propagation material.

Description

Technical Field

[0001] This invention belongs to the field of plant protection, specifically relating to methods for controlling or preventing pathogenic microorganisms on useful plants, particularly species of the genus *Fusarium* such as *Fusarium sp.*, *Fusarium graminearum*, *Ustilaginoideavirens*, *Clavicepssorghi*, *Fusarium culmorum*, *Botrytis cinerea*, *Phytophthora sojae*, *Phytophthora capsici*, *Phytophthora infestans*, *Exserohilumturcicum*, *Helminthosporium maydis*, *Colletotrichum gloeosporioides*, *Colletotrichum truncatum*, and *Pythium*. Methods of infection by *Deliense Meurs*, *Puccinia striiformis*, *Puccinia polysora Underw*, *Pseudoperonosporacubensis*, and *Erysiphe cichoracearum*. Background Technology

[0002] Species of the genus *Fusarium*, such as *Fusarium pseudograminearum*, *Fusarium culmorum*, *Fusarium graminearum*, *Fusarium auriumavenaceum*, *Fusarium moniliforme*, *Fusarium culmorum*, and *Fusarium acuminatum*, are fungal pathogens that cause wheat scab and wheat stem rot, especially *Fusarium graminearum*. If left uncontrolled, infected wheat exhibits reduced dry grain weight, decreased germination rate, weakened germination vigor, low flour yield, poor flour quality, dull color, and reduced commercial value. Furthermore, infected wheat contains toxins such as emetics and estrogen-like substances, which can cause acute poisoning in humans and animals. Diseased wheat grains contain toxins such as deoxynivalenol and zearalenone, which can cause pregnant animals to miscarry after poisoning. They can also infect cereal crops such as barley (Hordeum vulgare), rice (Oryza sativa), and oats (Arena sativa).

[0003] *Ustilaginoideavirens* is a fungal disease that causes rice false smut (*S. oryzae*). This disease occurs only in the panicle, affecting some of the grains. It initially appears as small, pale yellowish-green clumps at the glume's seam, gradually swelling and eventually enveloping the entire glume, turning dark green or olive-colored. Finally, it cracks, revealing a dark green powdery coating. Rice false smut is a major rice disease, distributed throughout rice-producing regions worldwide. The general incidence rate is 3-5%, but in severe cases it can exceed 30%, resulting in yield losses of 20-30%. It also produces rice blastotoxin, which is harmful to humans and animals. If left uncontrolled, rice false smut can lead to severe plant damage and yield reduction, and harm human and animal health.

[0004] Clavicepssorghi is a fungal disease that causes ergot disease in sorghum. The disease initially manifests as dark green, slightly wrinkled symptoms on the ovary, with white mycelial stroma at the base. It gradually spreads upwards, transforming the ovary into a fungal stroma. Five to ten days after infection, pink or light brown honeydew is secreted from the infected ovary. Currently, besides selecting resistant varieties, there are no other effective control measures. If left uncontrolled, sorghum ergot disease can lead to severe plant losses and yield reduction.

[0005] Botrytis cinerea is a omnivorous fungal pathogen that causes gray mold on tomatoes, and can also cause damping-off, leaf drop, flower rot, fruit rot, and cellar rot in seedlings, fruits, and storage organs of various other plants such as eggplant, lettuce, spinach, cucumber, bell pepper, strawberry, Malabar spinach, green beans, and lettuce. If left uncontrolled, Botrytis cinerea can lead to severe plant losses and yield reductions.

[0006] Phytophthora sojae is an oomycete disease that causes soybean phytosis, infecting soybeans at all stages of their growth. It is typically more severe in humid, rainy weather conditions. The disease has a short incubation period, allows for multiple reinfections, and spreads rapidly, causing widespread death of host plants. Without control, soybean phytosis can lead to severe soybean losses and yield reductions.

[0007] Phytophthora capsici is an oomycete pathogen that causes the causal disease of pepper blight. The disease can occur at both the seedling and mature plant stages, but is more prevalent in mature plants. The pathogen infects roots, stems, leaves, and fruits, making it a devastating disease in greenhouse pepper production. Severe outbreaks often result in total crop failure and can also damage tomatoes, eggplants, and cucurbits. Without control, Phytophthora capsici can lead to severe plant losses and yield reductions.

[0008] Phytophthora infestans is an oomycete pathogen that causes potato late blight. This disease primarily affects potato stems, leaves, and tubers, but can also infect flower buds and berries. If left uncontrolled, potato late blight can lead to severe plant losses and yield reductions.

[0009] *Exserohilum turcicum* is a fungal pathogen that infects and damages maize, causing maize leaf spot disease. The pathogen infects maize by producing appressoriums, special cell structures that use turgor pressure and mechanical force to penetrate the host plant's epidermis, thus impacting yield. If left uncontrolled, maize leaf spot disease can lead to severe plant losses and yield reductions.

[0010] Helminthosporium maydis is a fungal pathogen that causes short leaf spot disease in maize. It primarily affects maize leaves, but leaf sheaths, husks, and ears can also be infected. It can occur throughout the entire growth period of maize, but is most severe during the tasseling and grain-filling stages. If left uncontrolled, short leaf spot disease can lead to serious plant losses and yield reductions.

[0011] Colletotrichum gloeosporioides is a fungal pathogen that causes anthracnose in apples, primarily affecting the fruit but also infecting the fruit stalks and branches. Besides apples, it can also infect a wide variety of fruit trees, including crabapple, pear, grape, peach, walnut, hawthorn, persimmon, jujube, chestnut, citrus, lychee, and other fruits, as well as trees such as black locust. If left uncontrolled, apple anthracnose can lead to severe plant losses and yield reductions.

[0012] Colletotrichum truncatum is a fungal pathogen that causes anthracnose in soybeans. The disease can occur from the seedling stage to maturity, affecting cotyledons, leaves, petioles, stems, pods, and seeds. Soybean anthracnose is one of the most important diseases affecting soybeans worldwide, and if left uncontrolled, it can cause severe yield losses.

[0013] Pythium deliense Meurs is an oomycete pathogen that causes cottony rot in peppers. This disease leads to basal rot in seedlings, causing them to constrict and die. In mature plants, it primarily affects the fruit. The diseased parts of the fruit develop a brown, wet rot; under high humidity, a dense, white, cottony mold layer grows on the affected area, and in severe cases, the entire fruit rots. If left uncontrolled, cottony rot in peppers can cause significant plant losses and yield reduction.

[0014] Puccinia striiformis is a fungal pathogen that causes stripe rust on wheat. The disease can occur from wheat emergence to maturity, primarily affecting the leaves, followed by leaf sheaths and stems, and also infecting the ears, glumes, and awns. Wheat stripe rust is one of the most important wheat diseases worldwide and a significant biological hazard affecting wheat production in China. In years of severe outbreaks, it can lead to yield losses of over 40%, or even total crop failure.

[0015] Puccinia polysora Underw. is a fungal pathogen that causes southern rust in maize. It primarily affects the leaves, but can also infect leaf sheaths, stems, and husks. If left uncontrolled, southern rust can lead to severe plant losses and yield reductions.

[0016] Pear rust (Gymnosporangium haraeanum Syd.) is a fungal pathogen that causes rust disease in pear trees. It primarily affects leaves and new shoots, and in severe cases, also damages young fruit, petioles, and fruit stalks. Pear rust is a significant disease of pear trees; in severe years, the disease incidence rate of infected pear varieties in some orchards can exceed 60%. If left uncontrolled, pear rust can lead to severe plant losses and yield reduction.

[0017] Pseudoperonosporacubensis is an oomycete pathogen that causes cucumber downy mildew. After infection, it can cause most of the cucumber leaves to wither and die within one to two weeks, turning the cucumber field yellow and rotten. If left uncontrolled, cucumber downy mildew can lead to severe plant losses and yield reduction.

[0018] Erysiphe cichoracearum is a fungal pathogen that causes powdery mildew on cucumbers. The disease typically worsens in the later stages of growth, causing leaves to wither and even premature vine death. Besides cucumbers, it can also damage zucchini, pumpkins, and melons. If left uncontrolled, cucumber powdery mildew can lead to severe plant losses and yield reduction.

[0019] These diseases have a detrimental impact on agricultural output, and therefore there is a need to provide effective alternatives to common practices for controlling or preventing these diseases in vegetable, fruit, and grain plants. Therefore, this invention further provides a method for controlling or preventing infection of vegetable, fruit, and grain plants by plant pathogenic microorganisms that cause diseases such as wheat scab, wheat stem rot, rice false smut, gray mold of tomatoes and strawberries, soybean blight, pepper blight, potato late blight, corn large leaf spot, corn small leaf spot, apple and grape anthracnose, pepper cottony rot, wheat stripe rust, southern rust of corn, pear rust, cucumber downy mildew, and cucumber powdery mildew.

[0020] Patents CN113683509A and CN114890899A disclose a type of diphenyl ether ester compound of formula (II) and its preparation method, and disclose its application in controlling rice blast. However, no reports have been found on the control methods of other plant diseases using the diphenyl ether ester compounds of formula (II) specifically disclosed in patents CN113683509A and CN114890899A.

[0021]

[0022] Most pathogenic fungi or oomycetes rely on asexual conidia or zoospores to infect and spread diseases. If the germination of pathogen spores can be inhibited, plant infection by pathogenic microorganisms can be controlled or prevented. Summary of the Invention

[0023] The technical solution of the present invention to solve the above-mentioned technical problems is as follows:

[0024] This invention provides a method for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, wherein the plant pathogenic microorganisms are selected from Fusarium pseudograss, Fusarium xanthophytum, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora caulis, Helicobacter umbellatus, Helicobacter lanceolata, Anthracnose glomeratus, Pythium delavayi, Pseudomonas spp. ... and Powdery mildew. The method comprises applying a compound of formula (I) or its agriculturally active salt to the plant, its location, or its propagation material.

[0025]

[0026] Preferably, the useful plants are selected from wheat, barley, rice, sorghum, oats, tomatoes, strawberries, grapes, soybeans, peppers, potatoes, corn, apples, cucumbers, melons, okra, spinach, lettuce, asparagus, cabbage, carrots, onions, peppers, pears, peaches, walnuts, hawthorns, persimmons, dates, chestnuts, citrus fruits, lychees, zucchini, pumpkins, and cotton.

[0027] Furthermore, the plant pathogenic microorganisms mentioned herein are selected from Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora virosa, Helicobacter spp., Colletotrichum spp., Pythium driezii, Pseudomonas spp., Pseudomonas cucumeroides, Fusarium oxysporum, Helicobacter spp., Pseudomonas spp., Phytophthora virosa, Pseudomonas spp., Pseudomonas spp., Pseudomonas spp., Pseudomonas spp., Pseudomonas spp., Pseudomonas spp., Pseudomonas spp., and Powdery mildew, and the plants mentioned herein are selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

[0028] More preferably, the plant pathogenic microorganism is Fusarium graminearum, and the plant is wheat.

[0029] The present invention also provides the use of a compound of formula (I) or its agrochemically active salt for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, wherein the plant pathogenic microorganisms are selected from Fusarium pseudograss, Fusarium xanthophytum, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora caulis, Helicobacter umbellatus, Helicobacter spp., Colletotrichum gloeosporioides, Colletotrichum gloeosporioides, Pythium delavayi, Pseudomonas spp. ...

[0030] Preferably, the plant is selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

[0031] The present invention also provides a method for growing useful plants, the method comprising treating its propagation material with a compound of formula (I) or its agrochemically active salt, for controlling or preventing the useful plants from being infected by plant pathogenic microorganisms, said plant pathogenic microorganisms being selected from Fusarium pseudograss, Fusarium xanthophytum, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora caulis, Helicobacter umbellatus, Helicobacter lanceolata, Anthracnose glomeratus, Pythium delavayi, Pseudomonas spp. ... and Powdery mildew.

[0032] Preferably, the plant is selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

[0033] The present invention also provides the use of the compound of formula (I) or its agrochemically active salt in the preparation of a medicament for controlling or preventing useful plants from being infected by plant pathogenic microorganisms, wherein the plant pathogenic microorganisms are selected from Fusarium pseudograss, Fusarium xanthophytum, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora virosa, Helicobacter umbellatus, Helicobacter spp., Colletotrichum gloeosporioides, Colletotrichum gloeosporioides, Pythium delavayi, Pseudomonas spp. ...

[0034] Preferably, the plant is selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

[0035] Therefore, the compound of formula (I) provides farmers with an important new solution for controlling or preventing diseases in vegetables, fruits and grains such as wheat scab, rice false smut, sorghum ergot, wheat stem base rot, tomato and strawberry gray mold, soybean blight, pepper blight, potato late blight, maize large leaf spot, maize small leaf spot, apple and grape anthracnose, pepper cottony rot, wheat stripe rust, maize southern rust, pear rust, cucumber downy mildew and cucumber powdery mildew.

[0036] Compound (I) was optimized based on the skeleton of compound (II), which is an inhibitor designed to target the MoErs1 effector protein specific to rice blast fungus. This target is not found in other plant pathogens. The optimized compound (I) has unpredictable control effects on diseases or pathogens with large differences. Surprisingly, compound (I) has been found to be highly effective in controlling or preventing plant infection by plant pathogens, especially Fusarium graminearum, Fusarium graminearum, Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani, Ergotia spp., Botrytis cinerea, Phytophthora spp., Phytophthora capsici, Phytophthora virosa, Helicobacter umbellatus, Helicobacter lanceolata, Anthracnose, Anthracnose scutellaria, Pythium driezii, Pseudomonas spp. ...

[0037] Compounds of formula (I) can be used on their own or as agriculturally active salts. Agriculturally active salts include base addition salts of inorganic and organic bases. Examples include potassium salts, sodium salts, ammonium salts, dimethylamine salts, and isopropylamine salts.

[0038] The term "control" or "controlling" as used in this article includes protective, therapeutic, and eradicative treatments against plant pathogenic fungi.

[0039] Applying to plants should be understood as meaning applying to all above-ground and below-ground parts and organs of a plant, such as buds, leaves, needles, stems, trunks, flowers, fruiting bodies, fruits, seeds, roots, tubers, and rhizomes. It also includes applying to harvested material as well as asexual and sexually propagated material, such as cuttings, tubers, rhizomes, divisions, and seeds.

[0040] The plant pathogenic fungi that can be controlled according to the present invention include:

[0041] Powdery mildew pathogens include species of the genus *Blumeria*, such as *Blumeria graminis* (of the Poaceae family); species of the genus *Podosphaera*, such as *Podosphaeraleucotricha*; species of the genus *Sphaerotheca*, such as *Sphaerotheca fuliginea*; and species of the genus *Uncinula*, such as *Erysiphenecator* (the causal agent of grape powdery mildew).

[0042] Rust pathogens include species of the genus *Gymnosporangium*, such as *Gymnosporangium sabinae*; species of the genus *Hemileia*, such as *Hemileia vastatrix*; species of the genus *Phakopsora*, such as *Phakopsora pachyrhizi*, *Phakopsora meibomiae*, or *Phakopsora euvitis*; species of the genus *Puccinia*, such as *Puccinia recondita*, *Puccinia graminis*, or *Puccinia striiformis*; and species of the genus *Uromyces*, such as *Uromyces appendiculatus*.

[0043] Leaf blotch pathogens and leaf wilt pathogens include: species of the genus *Alternaria*, such as *Alternaria solani*; species of the genus *Cercospora*, such as *Cercosporabeticola*; species of the genus *Cladiosporum*, such as *Cladiosporium cucumerinum*; species of the genus *Cochliobolus*, such as *Cochliobolus sativus* (conidia form: *Drechslera*, synonym: *Helminthosporium*) or *Cochliobolus miyabeanus*; and species of the genus *Colletotrichum*, such as *Colletotrichum beanus*. * *Lindemuthanium*; species of the genus *Corynespora*, such as *Corynesporacassiicola*; species of the genus *Cycloconium*, such as *Cycloconium oleaginum*; species of the genus *Diaporthe*, such as *Diaporthecitri*; species of the genus *Elsinoe*, such as *Elsinoefaw*. cettii); Species of the genus *Gloeosporium*, such as *Gloeosporium laeticolor*; Species of the genus *Glomerella*, such as *Glomerella cingulata*; Species of the genus *Guignardia*, such as *Guignardia bidwelli*; Species of the genus *Leptosphaeria*, such as *Leptosphaeria maculans*; Species of the genus *Ustilaginoidea*, such as *Ustilaginoideavirens*; Species of the genus *Claviceps*, such as *Claviceps sorghi*; Species of the genus *Magnaporthe*, such as *Magnaporthe grisea*; Species of the genus *Microdochium*, such as *Microdochium nivale*.Species of the genus *Mycosphaerella*, such as *Zymoseptoriatritici*, *Mycosphaerella aracidola*, or *Mycosphaerella fijiensis*; species of the genus *Phaeosphaeria*, such as *Phaeosphaerianodorum*; species of the genus *Pyrenophora*, such as *Pyrenophorateres* or *Pyrenophora triticirepentis*; and species of the genus *Ramularia*, such as *Ramularia simja*. * *Collo-cygni* or *Ramularia areola*; species of the genus *Rhynchosporium*, such as *Rhynchosporium secalis*; species of the genus *Septoria*, such as *Septoria apii* or *Septorialycopersii*; species of the genus *Stagonospora*, such as *Stagonosporanodorum*; species of the genus *Typhula*, such as *Typhula incarnata*; species of the genus *Venturia*, such as *Venturia inaequalis*.

[0044] Diseases of the ear or panicle (including maize ears and cobs) caused by the following pathogens: species of the genera *Alternaria*, such as *Alternaria spp.*; species of the genera *Aspergillus*, such as *Aspergillus flavus*; species of the genera *Cladosporium*, such as *Cladosporium cladosporioides*; species of the genera *Claviceps*, such as *Claviceps purpurea*; species of the genera *Fusarium*, such as *Fusarium culmorum*; species of the genera *Gibberella*, such as *Gibberella zeae*; species of the genera *Monographella*, such as *Monographella nivalis*; and species of the genera *Cellatocystis*, such as the wheat leaf spot fungus.

[0045] Fruit rot can be caused by pathogens such as: species of the genus *Aspergillus*, such as *Aspergillus flavus*; species of the genus *Botrytis*, such as *Botrytis cinerea*; species of the genus *Monilinia*, such as *Monilinialaxa*; species of the genus *Penicillium*, such as *Penicillium expansum* or *Penicillium purpurogenum*; species of the genus *Rhizopus*, such as *Rhizopus stolonifer*; species of the genus *Sclerotinia*, such as *Sclerotinias clerotiorum*; and species of the genus *Verticilium*, such as *Verticilium alboatrum*.

[0046] Seed-borne and soil-borne rot and wilting diseases, as well as seedling diseases, are caused by the following pathogens: species of the genus *Alternaria*, such as *Alternaria brassicicola*; species of the genus *Aphanomyces*, such as *Aphanomyces euteiches*; species of the genus *Ascochyta*, such as *Ascochytalentis*; species of the genus *Aspergillus*, such as *Aspergillus flavus*; species of the genus *Cladosporium*, such as *Cladosporium herbarum*; and species of the genus *Cochliobolus*, such as *Cochliobolus granatum*. (sativus) (conidia: Drechslera, Bipolaris; synonym: Helminthosporium); species of the genus *Colletotrichum*, such as *Colletotrichum coccodes*; species of the genus *Fusarium*, such as *Fusarium luteum*. * *Culmorum*; species of the genus *Gibberella*, such as *Gibberella zeae*; species of the genus *Macrophomina*, such as *Macrophomina phaseolina*; species of the genus *Microdochium*, such as *Microdochium nivale*; species of the genus *Monographella*, such as *Monographella nivalis*; species of the genus *Penicillium*, such as *Penicillium expansum*; species of the genus *Phoma*, such as *Phoma lingam*; species of the genus *Phomopsis*, such as *Phomopsis sojae*; species of the genus *Phytophthora*, such as *Phytophthora malathi*. cactorum); species of the genus Pyrenophora, such as Pyrenophora graminea; species of the genus Pyricularia, such as Pyricularia oryzae; species of the genus Pythium, such as Pythium ultimum.Species of the genus *Rhizoctonia*, such as *Rhizoctonia solani*; species of the genus *Rhizopus*, such as *Rhizopus oryzae*; species of the genus *Sclerotium*, such as *Sclerotium rolfsii*; species of the genus *Septoria*, such as *Septorianodorum*; species of the genus *Typhula*, such as *Typhula incarnata*; and species of the genus *Verticillium*, such as *Verticillium dahliae*.

[0047] Diseases of plant tubers caused by the following pathogens: for example, species of the genus Rhizoctonia, such as Rhizoctonia solani; species of the genus Helminthosporium, such as Helminthosporium solani.

[0048] Diseases of soybean leaves, stems, pods, and / or seeds caused by the following pathogens: for example, Alternaria leaf spot (Alternaria spec.atranstenuissima) and anthracnose (Colletotrichum gloeosporoides dematium var.).truncatum, brown spot (Septoria glycines), Cercospora leaf spot and blight (Cercospora kikuchii), Choanephora leaf blight (Choanephora infundibulifera trispora), Dactuliophora leaf spot (Dactuliophoraglycines), downy mildew (Peronospora manshurica), drechslerablight (Drechsleraglycini), frogeye leaf spot (Cercospora sojina), Leptosphaerulina leaf spot (Leptosphaerulina trifolii), phyllostica leaf spot Spot (Phyllostictasojaecola)), pod and stem wilt (Phomopsis sojae)), powdery mildew (Microsphaeradiffusa), pyrenochaeta leaf spot (Pyrenochaetaglycines), rhizoctonia aerial, leaf blight and damping-off (Rhizoctonia solani)), rust (Phakopsorapachyrhizi, Phakoporameibomiae, Phakoporaeuvitis), scab (Sphacelomaglycines)), stemphylium leafblight (Stemphylium botryosum)), sudden death syndrome (Fusarium) Virguliforme), target spot disease (Corynesporacassiicola).

[0049] Preferably, the plant pathogenic fungi are selected from *Sclerotium spp.*, *Botrytis cinerea*, *Pyrrosia spp.*, *Ergacotyle spp.*, *Syngonium spp.*, *Cercospora spp.*, *Cercospora spp.*, *Rhizoctonia solani*, *Laminaria japonica*, *Cyclocarya spp.*, *Phyllostachys spp.*, *Phyllostachys spp.*, *Phyllostachys spp.*, *Phyllostachys spp.*, *Alternaria spp.*, *Cercospora spp.*, *Guignardia bidwellii*, *Sclerotium spp.*, *Microcystis spp.*, *Triticum aestivum*, *Cercospora spp.*, *Sclerotium ...

[0050] More preferably, the plant pathogenic fungi are selected from *Fusarium graminearum*, *Fusarium xanthosporum*, *Fusarium oxysporum*, *Fusarium graminearum*, *Rhizoctonia solani*, *Botrytis cinerea*, *Phytophthora sacchariformis*, *Phytophthora capsici*, *Phytophthora virosa*, *Helicobacter fusiformis*, *Colletotrichum gloeosporioides*, *Colletotrichum gloeosporioides*, *Pythium delavayi*, *Russula styracifolium*, *Russula multidus*, *Russula pear*, *Pseudomonas columbarum*, and *Phenyleton heliotropium*. *Fusarium graminearum* is particularly preferred.

[0051] Useful plants include, but are not limited to:

[0052] Vegetables or vegetable plants: including but not limited to okra, spinach, lettuce, asparagus, cabbage, carrots, onions, peppers, chili peppers, bell peppers, cucumbers, corn, lettuce, asparagus, bok choy, tomatoes, pumpkin, eggplant, and beets;

[0053] Fruits or fruit plants: including but not limited to grapes, apples, pears, peaches, hawthorns, persimmons, dates, citrus fruits, cherries, strawberries, blueberries, oranges, lemons, grapefruits, plums, apricots, bananas, sugarcane, and lychees;

[0054] Grains or grain plants: including but not limited to wheat, barley, rye, rice, oats, sorghum, soybeans and potatoes;

[0055] Oilseed crops: rapeseed, mustard, poppy, olive, sunflower, coconut, castor oil plants, cocoa and peanut;

[0056] Other plants: turf, tobacco, nuts, coffee, tea, pepper, grapevines, hops, and latex plants;

[0057] Ornamental plants: flowers, shrubs, deciduous trees and conifers.

[0058] Plant pathogenic microorganisms include fungi resistant to other fungicides. A fungus "resistant" to a specific fungicide is, for example, a strain that is less sensitive to that fungicide compared to the expected sensitivity of the same species. Expected sensitivity can be measured, for example, using strains that have not previously been exposed to the fungicide.

[0059] The method or use described above is preferably applied to the plant crop, its site, or its propagation material. Application is preferably to the plant site or the plant's propagation material, more preferably to the propagation material. Application can be carried out according to any common application method (e.g., foliar application, spraying, soil application, furrow irrigation, seed treatment, etc.).

[0060] Compound of formula (I) is preferably used for disease control at a concentration of 1 to 500 g / ha, more preferably 50 to 200 g / ha.

[0061] Compounds of formula (I) are suitable for use on any vegetable, fruit, or grain plant, including those useful plants that have been genetically modified to be resistant to active ingredients (such as herbicides) or those useful plants that have been genetically modified to produce bioactive compounds that control infection by plant pathogenic microorganisms.

[0062] Compounds of formula (I) are typically used in the form of compositions containing a carrier (e.g., formulations). Compounds of formula (I) and their compositions can be used in various forms, such as aerosol sprayers, capsule suspensions, cold atomized concentrates, pulverizable powders, emulsifiable concentrates, oil-in-water emulsions, water-in-oil emulsions, encapsulated granules, fine granules, flowable concentrates for seed treatment, gases (under pressure), gas-producing products, granules, thermal atomized concentrates, large granules, microgranules, oil-dispersible powders, oil-miscible flowable concentrates, oil-miscible liquids, pastes, plant sticks, powders for dry seed treatment, seeds coated with pesticides, soluble concentrates, soluble powders, solutions for seed treatment, suspension concentrates (flowable concentrates), ultra-low volume (ulv) liquids, ultra-low volume (ulv) suspensions, water-dispersible granules or tablets, water-dispersible powders for slurry treatment, water-soluble granules or tablets, water-soluble powders and wettable powders for seed treatment.

[0063] Formulations typically comprise a liquid or solid carrier and optionally one or more commonly used formulation aids, which may be solid or liquid aids, such as non-epoxidized or epoxidized vegetable oils (e.g., epoxidized coconut oil, rapeseed oil, or soybean oil), defoamers (e.g., silicone oil), preservatives, clay, inorganic compounds, viscosity modifiers, surfactants, binders, and / or thickeners. The composition may further comprise fertilizers, micronutrient donors, or other products that influence plant growth, and may include combinations containing the compounds of the present invention and one or more other bioactive agents, such as bactericides, fungicides, nematicides, plant activators, acaricides, and insecticides.

[0064] The composition is prepared by methods known per se, in the absence of adjuvants, for example by grinding, sieving, and / or compressing the solid compound of the invention, and in the presence of at least one adjuvant, for example by tightly mixing and / or grinding the compound of the invention together with one or more adjuvants. In the case of the solid compound of the invention, the grinding / crushing of the compound is to ensure a specific particle size.

[0065] Examples of compositions for use in agriculture are emulsifiable concentrates, suspension concentrates, microemulsions, oil dispersibles, direct sprayable or dilutable solutions, coatable pastes, diluted emulsions, soluble powders, dispersible powders, wettable powders, dusts, granules, or capsules in polymeric substances, wherein the composition comprises a compound of formula (I) and the type of composition is selected to suit the intended purpose and the prevailing environment.

[0066] Typically, the composition comprises 0.1% to 99% (especially 0.1% to 95%) of a compound of formula (I) and 1% to 99.9% (especially 5% to 99.9%) of at least one solid or liquid carrier. It is also possible, in principle, that 0% to 25% (especially 0.1% to 20%) of the composition is a surfactant (in each case, % refers to weight percentage). For commercial purposes, concentrated compositions are preferred, and end users typically use diluted compositions with substantially lower concentrations of active ingredient.

[0067] Examples of leaf formulations used in premixed compositions are:

[0068] GR: Granules

[0069] WP: Wettable powder

[0070] WG: Water-dispersible granules (powders)

[0071] SG: Water-soluble granules

[0072] SL: Soluble Concentrate

[0073] EC: Emulsifiable concentrate

[0074] EW: Oil-in-water emulsion

[0075] ME: Microemulsion

[0076] SC: Aqueous suspension concentrate

[0077] CS: Aqueous Capsule Suspension

[0078] OD: Oil-based suspension concentrate, and

[0079] SE: Aqueous suspension emulsion.

[0080] Examples of seed-treated formulations used in premixed compositions include:

[0081] WS: Wettable powder for seed treatment slurry

[0082] LS: Solution for seed treatment

[0083] ES: Emulsion for seed treatment

[0084] FS: Suspension concentrate for seed treatment

[0085] WG: Water-dispersible granules, and

[0086] CS: Aqueous capsule suspension.

[0087] Examples of formulation types suitable for barrel-mixing compositions include solutions, diluted emulsions, suspensions or mixtures thereof, and dusting agents.

[0088] The application method (such as foliar application, spraying, misting, atomizing, dusting, broadcasting, coating, or dumping) can be selected according to the intended purpose and the prevailing environment. Barrel-mixed compositions are generally prepared by diluting one or more premixed compositions containing different biocides and optionally additional adjuvants with a solvent (e.g., water).

[0089] Suitable carriers and auxiliaries can be solid or liquid and are substances commonly used in formulation technology, such as natural or recycled minerals, solvents, dispersions, wetting agents, thickeners, binders, or fertilizers.

[0090] In general, the tank-mixed product for foliar or soil application contains 0.1% to 20%, especially 0.1% to 15%, of the desired ingredient and 99.9% to 80%, especially 99.9% to 85%, of solid or liquid adjuvants (including, for example, solvents such as water), wherein the adjuvant may be a surfactant, in an amount based on 0 to 20%, especially 0.1% to 15%, of the tank-mixed product.

[0091] Typically, premixed formulations for foliar application contain 0.1% to 99.9%, particularly 1% to 95%, of the desired ingredient and 99.9% to 0.1%, particularly 99% to 5%, of solid or liquid adjuvants (including, for example, solvents such as water), wherein the adjuvants may be surfactants, in amounts based on 0% to 50%, particularly 0.5% to 40%, of the premixed formulation.

[0092] Typically, tank-mixed products for seed treatment application contain 0.25% to 80%, especially 1% to 75%, of the desired ingredient and 99.75% to 20%, especially 99% to 25%, of solid or liquid adjuvants (including, for example, solvents such as water), wherein the adjuvants may be surfactants, in amounts based on 0% to 40%, especially 0.5% to 30%, of the tank-mixed product.

[0093] Typically, premixed preparations for seed treatment application contain 0.5% to 99.9%, especially 1% to 95% of the desired ingredient and 99.5% to 0.1%, especially 99% to 5% of solid or liquid adjuvants (including, for example, solvents such as water), wherein the adjuvants may be surfactants, in amounts based on 0% to 50%, especially 0.5% to 40% of the premixed preparation.

[0094] Commercial products will preferably be formulated as concentrates (e.g., premixed compositions (formulations)), while end users will typically use diluted formulations (e.g., barrel-mixed compositions).

[0095] Preferred seed treatment premixes are aqueous suspension concentrates. The formulation can be applied to seeds using conventional processing techniques and machinery, such as fluidized bed technology, drum milling methods, rotostatic seed processors, and rotary drum coaters. Other methods, such as spray beds, can also be useful. Seeds can be pre-sized before coating. After coating, the seeds are typically dried and then transferred to a sizing machine for sizing. Such procedures are known in the art. The compounds of the present invention are particularly suitable for soil and seed treatment applications.

[0096] In general, the premixed composition of the present invention contains 0.5% to 99.9%, especially 1% to 95%, advantageously 1% to 50% by weight of the desired component and 99.5% to 0.1%, especially 99% to 5% by weight of solid or liquid adjuvants (including, for example, solvents such as water), wherein the adjuvants (or auxiliary agents) may be surfactants, and the amount thereof is 0% to 50%, especially 0.5% to 40% by weight based on the mass of the premixed formulation.

[0097] Plants and cultivars treated with compounds of formula (I) include plants and cultivars that are hybrid plants that have expressed characteristics of heterosis or hybrid vigor, which generally result in higher yield, vigor, health and resistance to biotic and abiotic stresses.

[0098] Compounds of formula (I) can be advantageously used to treat transgenic plants, cultivars, or plant parts that have received genetic material that confers advantageous and / or useful characteristics (traits) to these plants, cultivars, or plant parts. Therefore, it is conceivable to combine the present invention with one or more recombinant traits or transgenic events, or combinations thereof. For the purposes of this application, a transgenic event is generated by inserting a specific recombinant DNA molecule into a specific location (site) on a plant genome chromosome. This insertion creates a new DNA sequence referred to as an "event," characterized by the inserted recombinant DNA molecule and a certain amount of genomic DNA adjacent to / flanking both ends of the inserted DNA. Such traits or transgenic events include, but are not limited to, resistance to pests and diseases, water use efficiency, yield performance, drought tolerance, seed quality, improved nutritional quality, hybrid seed production, and herbicide tolerance, wherein said trait is determined relative to plants lacking such trait or transgenic event. Specific examples of such advantageous and / or useful traits are better plant growth, vigor, stress tolerance, stand growth capacity, lodging resistance, nutrient uptake, plant nutrition and / or yield, especially improved growth, increased tolerance to high or low temperatures, increased tolerance to drought or water or soil salinity levels, enhanced flowering performance, easier harvesting, accelerated ripening, higher yield, higher quality and / or higher nutritional value of harvested products, better storage life and / or processability of harvested products, and enhanced resistance to animal and microbial pests (e.g., insects, arachnids, nematodes, mites, slugs and snails).

[0099] Among the DNA sequences encoding proteins that confer tolerance traits to these animal and microbial pests (especially insects), particular mention will be made of the genetic material encoding Bt proteins from Bacillus thuringiensis, which is widely documented and well known to those skilled in the art. Proteins extracted from bacteria, such as those from the genus Photorhabdus (WO97 / 17432 and WO98 / 08932), will also be mentioned. In particular, VIP proteins that refer to BtCry or include CryA, CryIAb, CryIAc, CryIIA, CryIIIA, CryIIIB2, Cry9c, Cry2Ab, Cry3Bb and CryIF proteins or their toxic fragments, as well as their hybrids or combinations thereof, especially CryF proteins or hybrids derived from CryF proteins (such as hybrid CryA-CrylF proteins or their toxic fragments), CryA-type proteins or their toxic fragments, preferably CryAc proteins or hybrids derived from CryAc proteins (such as hybrid CryAb-CrylAc proteins) or CryAb or Bt2 proteins or their toxic fragments, Cry2Ae, Cry2Af or Cry2Ag proteins or their toxic fragments, CryA.105 proteins or their toxic fragments, VIP3Aa19 proteins, VIP3Aa20 proteins, and VIP3A proteins generated in the COT202 or COT203 cotton events, are described in Estruch et al. (1996), Proc Natl Acad. The VIP3Aa protein or its toxic fragment as described in Sci US A.28;93(11):5389-94, the Cry protein described in WO2001 / 47952, and insecticidal proteins from strains of the genus Xenorhabdus (as described in WO98 / 50427), the genus Serratia (particularly from S. entomophila), or the species of luminescent bacteria, such as the Tc protein from luminescent bacteria described in WO98 / 08932. Furthermore, this document also includes any variants or mutants of any of the aforementioned proteins that differ from any of the sequences (particularly the sequences of their toxic fragments) at some amino acid (1-10, preferably 1-5), or any variants or mutants of the aforementioned proteins fused with a transport peptide (such as a plastid transport peptide) or another protein or peptide.

[0100] The beneficial effects of this invention are as follows: the control effect of the same compound on diseases or pathogens with large differences is unpredictable. Unexpectedly, it has been found that the compound of formula (I) has a good control or prevention effect on useful plants infected by plant pathogenic microorganisms. Detailed Implementation

[0101] The present invention is illustrated below with reference to examples, but is not intended to limit the invention. Simple substitutions or modifications made to the present invention by those skilled in the art are within the scope of the technical solutions protected by this invention. The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values; these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0102] Several methods for preparing compounds of formula (I) of the present invention have been disclosed in CN113683509A, which is incorporated herein by reference. The starting materials can be commercially available or prepared using methods known in the literature. Those skilled in the art will understand that the compounds of the present invention can also be synthesized using other synthetic routes.

[0103] All plant pathogenic microorganisms in the biological examples were strains preserved at 4°C in the laboratory of the College of Plant Protection, Nanjing Agricultural University.

[0104] Water agar medium: Add 20g of agar to 1000mL of distilled water, stir to dissolve completely, dispense into 200mL Erlenmeyer flasks, sterilize at 121℃ for 20 minutes, and cool before use.

[0105] PDA medium (potato agar-glucose medium): Wash and peel 200g of potatoes, cut them into small pieces, boil them until soft (boil for 30 minutes), filter through four layers of gauze to remove the residue, add 20g of glucose, then add 20g of agar, add double-distilled water to make up to 1000mL, stir to dissolve completely, dispense into 200mL Erlenmeyer flasks, sterilize at 121℃ for 20 minutes, and cool before use.

[0106] V8 medium (10%): Add 1g CaCO3 to 100mL of V8 juice, centrifuge at 2500rpm for 5min, and dilute the supernatant with double-distilled water at a ratio of 1:9 (v / v). If preparing solid medium, add 15g agar powder to 1L of the diluted solution, sterilize at 121℃ for 20min, and cool before use.

[0107] PDAG medium: Wash and peel 200g of potatoes, cut them into small pieces, boil them until soft (boil for 30 minutes), filter them through four layers of gauze to remove the residue, add 20g of mannitol, then add 20g of agar, add double-distilled water to make up to 1000mL, stir to dissolve completely, dispense into 200mL Erlenmeyer flasks, sterilize at 121℃ for 20 minutes, and cool before use.

[0108] Medium concentration of EC for antibacterial 50Values, toxicity regression equations, and correlation coefficients are calculated using the DPS data processing system: input the actual test concentration and the corresponding spore germination inhibition rate, use quantitative biological value analysis to convert the measurement to logarithmic units, set the lethality rate value to 50, and obtain the required data.

[0109] The following examples are provided to further illustrate the preparation method of the present invention. The specific substances, types, and conditions used are intended to further explain the invention and are not intended to limit its reasonable scope. All referenced documents are incorporated herein by reference.

[0110] Formulation Example 1 (SC)

[0111] 10 parts of compound (I) (content >95%), 6 parts of wetting agent SP-SC3266E, 4 parts of ethylene glycol, 0.35 parts of xanthan gum, 0.2 parts of Kathon, 0.2 parts of defoamer, and 79.25 parts of water were mixed evenly, and then sheared, stirred, and milled to obtain an aqueous suspension concentrate containing 10% of the active ingredient.

[0112] Biological Example 1: Fusarium graminearum (wheat scab) on wheat

[0113] The spore germination assay was performed as follows: 100 mg of compound (I) (content >95%) was dissolved in 10 mL of DMSO to prepare a stock solution of 10000 μg / mL; different doses of the stock solution were added to PDA medium and diluted to different concentrations as shown in Table 1, and plates were inverted. The concentration of the conidial suspension was 1 × 10⁻⁶. 5 The spores were evenly spread onto plates of different concentrations using a spreader at a concentration of 0 μg / mL. 50 μL of spore suspension was added to each PDA plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination on the drug-free plates (0 μg / mL). When the spore germination rate on the drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment (number of germinating spores / 200) was recorded. The experiment was repeated three times, and the average value was taken. The inhibition of spore germination by compound (I) is shown in Tables 1 and 2.

[0114] Table 1 shows the inhibition rate (%) of compound (I) on spore germination of Fusarium graminearum.

[0115] 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 9.3 30.2 44.7 Carbendazim 7.3 23.3 36.2 Cyazofamid 8.1 26.7 39.6

[0116] Table 2 shows the inhibitory activity of compound (I) on spore germination of Fusarium graminearum.

[0117] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 3.6723 + 1.1824x 0.9997 13.2730 Carbendazim y = 3.5377 + 1.0895x 0.9990 21.9896 Cyazofamid y = 3.5985 + 1.1305x 0.9999 17.3696

[0118] Biological Example 2: Botrytis cinerea (Tomato gray mold) on tomatoes

[0119] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 3. The culture medium used for coating the spore suspension was water agar medium, and the spore suspension concentration was 1×10⁻⁶. 5 50 μL of spore suspension was added to each water agar plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on the drug-free plates. When the spore germination rate on the drug-free plates reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0120] Table 3 shows the inhibition rate (%) of compound (I) on spore germination of Botrytis cinerea.

[0121] 2μg / mL 10 μg / mL 20 μg / mL Compound of formula (I) 10.6 39.3 60.5 Pyraclostrobin 22.6 56.1 72.9 iprodione 17.1 48.3 69.2 pyrimethanil 10.4 35.1 58.2

[0122] Table 4 shows the inhibitory activity of compound (I) on spore germination of *Botrytis cinerea*.

[0123] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 3.2866 + 1.4937x 0.9982 14.029 Pyraclostrobin y = 3.8326 + 1.3502x 0.9993 7.3212 iprodione y = 3.6005 + 1.4248x 0.9966 9.6007 pyrimethanil y = 3.2826 + 1.4287x 0.9936 15.9250

[0124] Biological Example 3: Phytophthora sojae on soybeans (Phytophthora sojae pathogen)

[0125] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 5. The culture medium used for coating the zoospore suspension was V8 medium, and the spore suspension concentration was 1 × 10⁻⁶. 5 50 μL of spore suspension was dropped onto each V8 medium (10%) plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0126] Table 5 shows the spore germination inhibition rate (%) of compound (I) on bacteria.

[0127] 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 30.1 46.2 50.1 Metalaxyl 9.2 31.6 53.3 Dimethomorph 60.2 89.3 96.1

[0128] Table 6 shows the inhibitory activity of compound (I) on spore germination of Phytophthora soybeanis.

[0129] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 4.4865 + 0.5389x 0.9932 8.971 Metalaxyl y = 3.6454 + 1.3769x 0.9941 9.6350 Dimethomorph y = 5.2457 + 1.4870x 0.9988 0.6835

[0130] Biological Example 4: Phytophthora capsici, the pathogen of which grows on chili peppers (Phytophthora capsici)

[0131] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 7. The culture medium used for coating the zoospore suspension was V8 medium, and the spore suspension concentration was 1 × 10⁻⁶. 5 50 μL of spore suspension was dropped onto each V8 medium (10%) plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0132] Table 7 shows the inhibition rate (%) of compound (I) on spore germination of *Phytophthora capsici*.

[0133] 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 16.3 44.6 62.4 Metalaxyl 13.6 40.1 61.3 Dimethomorph 51.1 78.3 88.7

[0134] Table 8 shows the inhibitory activity of compound (I) on spore germination of *Phytophthora capsici*.

[0135] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 4.0062 + 1.2828x 0.9986 5.9526 Metalaxyl y = 3.8785 + 1.3552x 0.9953 6.7230 Dimethomorph y = 5.0138 + 1.1650x 0.9977 0.9731

[0136] Biological Example 5: Pathogenic Phytophthora on Potatoes (Potato Late Blight)

[0137] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 9. The culture medium used for coating the spore suspension was V8 medium, and the spore suspension concentration was 1 × 10⁻⁶. 5 50 μL of spore suspension was dropped onto each V8 medium (10%) plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0138] Table 9 shows the inhibition rate (%) of compound (I) on spore germination of pathogenic *Phytophthora infestans*.

[0139] 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 12.6 34.7 54.1 Metalaxyl 9.8 32.6 52.3 Fluopyram 13.4 37.3 56.1 Dimethomorph 40.3 77.6 89.2

[0140] Table 10 shows the inhibitory activity of compound (I) against spore germination of pathogenic *Phytophthora infestans*.

[0141] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 3.8315 + 1.2181x 0.9942 9.1052 Metalaxyl y = 3.6875 + 1.3250x 0.9965 9.7850 Fluopyram y = 3.8737 + 1.2365x 0.9963 8.1443 Dimethomorph y = 4.7483 + 1.4747x 0.9997 1.4814

[0142] Biological Example 6: Long-spore fungus on maize (maize leaf spot pathogen)

[0143] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 11. The culture medium used for coating the spore suspension was PDAG medium, and the spore suspension concentration was 1×10⁻⁶. 5 50 μL of spore suspension was added to each PDAG agar plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0144] Table 11 shows the inhibition rate (%) of compound (I) on spore germination of *Helicobacter pylori*.

[0145]

[0146]

[0147] Table 12 shows the inhibitory activity of compound (I) on spore germination of *Helicobacter pylori*.

[0148] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 4.6499 + 0.5106x 0.9989 4.850 Tebuconazole y = 4.4263 + 0.6524x 0.9949 7.5753 Pyraclostrobin y = 4.6859 + 1.0235x 0.9980 2.0273

[0149] Biological Example 7: Colloidal anthracnose on apples (Apple anthracnose pathogen)

[0150] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 13. The culture medium used for coating the spore suspension was PDA medium, and the spore suspension concentration was 1×10⁻⁶. 5 50 μL of spore suspension was added to each PDA medium plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor the spore germination process on the drug-free plates (0 μg / mL). When the spore germination rate on the drug-free plates (0 μg / mL) reached approximately 95%, the investigation began, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields of view were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0151] Table 13 shows the inhibition rate (%) of compound (I) on spore germination of *Colletotrichum gloeosporioides*.

[0152] 2μg / mL 10 μg / mL 20 μg / mL Compound of formula (I) 22.1 49.1 62.8 Imazalil 19.8 45.2 59.9 difenoconazole 37.2 65.1 77.9

[0153] Table 14 shows the inhibitory activity of compound (I) on spore germination of *Colletotrichum gloeosporioides*.

[0154] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 3.8992 + 1.0905x 0.9998 10.2196 Imazalil y = 3.8156 + 1.0894x 0.9992 12.2249 difenoconazole y = 4.3378 + 1.0825x 0.9987 4.0897

[0155] Biological Example 8: Pythium driezii on chili peppers (the fungus that causes cottony rot in chili peppers)

[0156] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 15. The culture medium used for coating the spore suspension was water agar medium, and the spore suspension concentration was 1×10⁻⁶. 5 50 μL of spore suspension was dropped onto each V8 medium (10%) plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0157] Table 15 shows the inhibition rate (%) of compound (I) on spore germination of *Pythium driezii*.

[0158] 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 73.7 82.2 91.1 Flupyrimethanil 45.6 66.2 76.2 iprodione 43.6 58.5 71.2

[0159] Table 16 shows the inhibitory activity of compound (I) on spore germination of *Pythium driezii*.

[0160]

[0161]

[0162] Biological Example 9: Fusarium oxysporum and Fusarium chrysogenum (wheat stem rot fungi) on wheat

[0163] The spore germination assay was performed as follows: 100 mg of compound (I) (content >95%) was dissolved in 10 mL of DMSO to prepare a stock solution of 10000 μg / mL; different doses of the stock solution were added to PDA medium and diluted to different concentrations as shown in the table, and plates were inverted. The concentration of the conidial suspension was 1 × 10⁻⁶. 5The spores were evenly spread onto plates of different concentrations using a spreader at a density of 0 μg / mL. 50 μL of spore suspension was added to each PDA plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination on the drug-free plates (0 μg / mL). When the spore germination rate on the drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment (number of germinating spores / 200) was recorded. The experiment was repeated three times, and the average value was taken.

[0164] The results showed that the compound of formula (I) had excellent antibacterial effects against Fusarium pseudograss and Fusarium xanthophyte.

[0165] Biological Example 10: Fusarium oxysporum (the causal agent of cotton wilt) on cotton

[0166] The spore germination assay was performed as follows: 100 mg of compound (I) (content >95%) was dissolved in 10 mL of DMSO to prepare a stock solution of 10000 μg / mL; different doses of the stock solution were added to PDA medium and diluted to different concentrations as shown in the table, and plates were inverted. The concentration of the conidial suspension was 1 × 10⁻⁶. 5 The spores were evenly spread onto plates of different concentrations using a spreader at a density of 0 μg / mL. 50 μL of spore suspension was added to each PDA plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination on the drug-free plates (0 μg / mL). When the spore germination rate on the drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment (number of germinating spores / 200) was recorded. The experiment was repeated three times, and the average value was taken.

[0167] The results showed that the compound of formula (I) had excellent antibacterial effect against Fusarium oxysporum.

[0168] Biological Example 11: Helicobacter convexum (maize leaf spot pathogen) on maize

[0169] The spore germination assay was performed: Compound (I) was diluted with DMSO to a stock solution of 10000 μg / mL, and the reagent concentrations were set as shown in Table 11. The culture medium used for coating the spore suspension was PDAG medium, and the spore suspension concentration was 1×10⁻⁶. 550 μL of spore suspension was added to each PDAG agar plate, spread evenly with a sterile glass rod, and incubated upside down at 25°C to monitor spore germination progress on drug-free plates (0 μg / mL). When the spore germination rate on drug-free plates (0 μg / mL) reached approximately 95%, an investigation was initiated, and all plates were placed at 0°C. During the investigation, 200 spores were counted in each field of view under a microscope (fields were not repeated), and the germination rate of each treatment was recorded; the experiment was repeated three times, and the average value was taken.

[0170] The results showed that the compound of formula (I) had excellent antibacterial effect against Helicobacter convexum.

[0171] summary:

[0172] As demonstrated by the above biological examples, compound (I) exhibits superior antifungal activity against *Fusarium graminearum* compared to the commercial standard fungicides carbendazim and cyazofamid; superior antifungal activity against *Botrytis cinerea* compared to the commercial standard fungicide pyrimethanil; superior antifungal activity against *Phytophthora spp.* and *Phytophthora capsici* compared to the commercial standard fungicide metalaxyl; comparable antifungal activity against pathogenic *Phytophthora* compared to the commercial standard fungicides metalaxyl and fluopyram; superior antifungal activity against *Helicobacter pylori* compared to the commercial standard fungicide tebuconazole; superior antifungal activity against *Colletotrichum gloeosporioides* compared to the commercial standard fungicide imazalil; and superior antifungal activity against *Pythium drizzii* compared to the commercial standard fungicides fluopyram and iprodione. Therefore, compound (I) shows significant inhibitory effects on the germination of spores of most pathogenic fungi or oomycetes, and has the function of controlling or preventing plant infection by pathogenic microorganisms.

[0173] Biological Example 12: Fusarium graminearum on wheat (causing wheat scab)

[0174] In a spraying chamber, flowering wheat (cultivar Zhengmai 0943) was sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (tebuconazole, cyazofamid, and carbendazim). The results were obtained by injecting spores (1×10⁻⁶) 24 hours after application. 5 The plants were inoculated with a suspension of 20 seedlings per mL (number of seedlings per mL) in both the control and treatment groups. The inoculated test plants were cultured in a greenhouse at 25°C and 95% relative humidity. 7-14 days after application, the number of diseased spikelets per plant was counted, and the inhibition rate was calculated as follows: Inhibition rate = (mean number of diseased spikelets in the control group - mean number of diseased spikelets in the treatment group) / mean number of diseased spikelets in the control group.

[0175] Table 17 shows the bioassay inhibition rate (%) of compound (I) against wheat scab.

[0176]

[0177]

[0178] Biological Example 13: Anthracnose fungus on soybeans (causing soybean anthracnose)

[0179] In a spraying chamber, soybean seedlings (cultivar Hefeng 47) at the 3-leaf stage were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (prochloraz, difenoconazole, and carbendazim). Conidia (1×10⁻⁶) were collected by dripping conidia 24 hours after application. 5 The plants were inoculated with a suspension of 20 seedlings each in the control and treatment groups. The inoculated test plants were cultured in a greenhouse at 90% relative humidity. When appropriate levels of lesions appeared on the untreated control plants (7-14 days after application), the leaf area affected was counted, and the inhibition rate was calculated as follows: Inhibition rate = (affected leaf area in the control group - affected leaf area in the treatment group) / affected leaf area in the control group.

[0180] Table 18 shows the inhibition rate (%) of compound (I) against soybean anthracnose.

[0181] 0 μg / mL 10 μg / mL 100 μg / mL 200 μg / mL 500 μg / mL Compound of formula (I) 0 21.6 53.3 69.8 87.6 Imazalil 0 23.3 50.8 68.6 85.6 difenoconazole 0 33.6 65.2 77.9 90.3 Carbendazim 0 13.9 41.3 62.1 76.2

[0182] Table 19 shows the inhibitory activity of compound (I) against soybean anthracnose.

[0183] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 3.1245 + 1.0316x 0.9910 65.7711 Imazalil y = 3.1610 + 1.0243x 0.9815 62.4367 difenoconazole y = 3.5296 + 0.9896x 0.9910 30.6117 Carbendazim y = 2.8067 + 1.0612x 0.9917 116.6434

[0184] Biological Example 14: Phytophthora infestans on potatoes (causing late blight of potatoes)

[0185] In a spraying chamber, potato seedlings (cultivar Fiurita) were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (metalaxyl, fluopyram, and dimethomorph). Twenty seedlings in each control and treatment group were inoculated by dripping a suspension of zoospores (50 spores / μL) 24 hours after application. The inoculated test plants were cultured in a greenhouse at 90% relative humidity. When appropriate levels of lesions appeared on the untreated control plants (7–14 days after application), the leaf area affected was counted, and the inhibition rate was calculated as: Inhibition rate = (Leaf area affected in control group - Leaf area affected in treatment group) / Leaf area affected in control group.

[0186] Table 20 shows the bioassay inhibition rate (%) of compound (I) against potato late blight.

[0187] 0 μg / mL 0.2 μg / mL 1μg / mL 2μg / mL 4μg / mL Compound of formula (I) 0 26.5 60.2 75.3 89.7 Metalaxyl 0 23.8 60.1 72.4 87.2 Fluopyram 0 29.1 63.4 77.8 91.3 Dimethomorph 0 16.8 42.8 58.3 72.9

[0188] Table 21 shows the inhibitory activity of compound (I) against potato late blight.

[0189] virulence regression equation Correlation coefficient <![CDATA[EC 50 (μg / mL)]]> Compound of formula (I) y = 5.3221 + 1.4235x 0.9953 0.5939 Metalaxyl y = 5.2474 + 1.3918x 0.9978 0.6641 Fluopyram y = 5.4060 + 1.4345x 0.9950 0.5211 Dimethomorph y = 4.8576 + 1.2019x 0.9989 1.3136

[0190] Biological Example 15: Stripe rust fungus on wheat (causing wheat stripe rust)

[0191] In a spraying chamber, three-leaf seedlings of wheat (cultivar Zhengmai 0943) were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (pyraclostrobin and difenoconazole). Spores (1×10⁻⁶) were sprayed 24 hours after application. 5 The plants were inoculated with a suspension of (number of plants / mL). The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When an appropriate level of disease appeared on the untreated control plants (7-14 days after application), the leaf area affected was counted and the inhibition rate was calculated as follows: Inhibition rate = (affected leaf area of ​​control group - affected leaf area of ​​treated group) / affected leaf area of ​​control group.

[0192] Table 22 shows the bioassay inhibition rate (%) of compound (I) against wheat stripe rust.

[0193] 0 μg / mL 0.2 μg / mL 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 0 26.5 62.9 78.3 89.7 Pyraclostrobin 0 23.8 61.1 76.4 87.2 difenoconazole 0 29.1 63.4 80.8 91.3

[0194] Biological Example 16: *Pseudomonas columbarium* on cucumbers (causing cucumber downy mildew)

[0195] In a spray chamber, four-leaf cucumber seedlings (cultivar Huai'an) were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (cypermethrin, dimethomorph, and carbendazim). Spores (1×10⁻⁶) were sprayed 24 hours after application. 5 The plants were inoculated with a suspension of (number of plants / mL). The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When appropriate levels of lesions appeared on the untreated control plants (7-14 days after application), the leaf lesion area was counted and the inhibition rate was calculated. Inhibition rate = (leaf lesion area of ​​control group - leaf lesion area of ​​treatment group) / leaf lesion area of ​​control group.

[0196] Table 23 shows the inhibition rate (%) of compound (I) against cucumber downy mildew.

[0197] 0 μg / mL 0.2 μg / mL 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 0 28.1 50.6 62.8 80.7 cyanazole 0 33.5 60.2 77.3 88.4 Dimethomorph 0 25.8 51.3 70.1 82.9 Carbendazim 0 21.6 40.7 56.3 68.4

[0198] Biological Example 17: Powdery mildew fungus on cucumbers (causing cucumber powdery mildew)

[0199] In a spraying chamber, cucumber leaves (cultivar Huai'an cucumber) at the 4-leaf stage were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (tebuconazole, flufenoxuron, and carbendazim). The concentration of conidia (1 × 10⁻⁶) was determined by spraying 24 hours after application. 5The plants were inoculated with a suspension of (number of plants / mL). The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When appropriate levels of lesions appeared on the untreated control plants (7-14 days after application), the leaf lesion area was counted and the inhibition rate was calculated. Inhibition rate = (leaf lesion area of ​​control group - leaf lesion area of ​​treatment group) / leaf lesion area of ​​control group.

[0200] Table 24 shows the inhibition rate (%) of compound (I) against cucumber powdery mildew.

[0201] 0 μg / mL 0.2 μg / mL 1μg / mL 5μg / mL 10 μg / mL Compound of formula (I) 0 24.8 48.5 62.3 73.2 Tebuconazole 0 22.6 46.3 58.6 70.1 Fluorophenyl ether amide 0 33.9 56.2 70.1 81.3 Carbendazim 0 14.3 27.9 41.9 59.4

[0202] Biological Example 18: Pear gum rust fungus on pear trees (causing pear gum rust disease)

[0203] In a spraying chamber, leaves of appropriately aged pear trees (pear cultivar: Su Cui No. 1) were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (pyraclostrobin and difenoconazole). The spores (1×10⁻⁶) were sprayed 24 hours after application. 5 The detached leaves were inoculated with a suspension of (number of leaves per mL). The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When an appropriate level of disease appeared on the untreated control plants (7-14 days after application), the diseased area of ​​the leaves was counted, and the inhibition rate was calculated as follows: Inhibition rate = (Diseased area of ​​control group leaves - Diseased area of ​​treated group leaves) / Diseased area of ​​control group leaves.

[0204] The results showed that compound (I) had excellent control efficacy against pear rust.

[0205] Biological Example 19: Rhizoctonia solani on rice (causing rice false smut)

[0206] In a spraying chamber, the compound of formula (I) prepared in Preparation Example 1 and the control agents (tebuconazole and difenoconazole) were sprayed onto rice panicles of appropriate age. The rice cultivar was Liangyoupei 9. Spores (1×10⁻⁶) were injected into the live rice panicles 24 hours after application. 5 A suspension of (number / mL) was prepared. The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When an appropriate level of disease appeared on the untreated control plants (7-14 days after application), the diseased area of ​​the panicles was counted, and the inhibition rate was calculated as follows: Inhibition rate = (Disease-affected area of ​​control group panicles - Disease-affected area of ​​treated group panicles) / Disease-affected area of ​​control group panicles.

[0207] The results showed that compound (I) had excellent control efficacy against rice false smut.

[0208] Biological Example 20: Ergot fungus on sorghum (causing ergot disease in sorghum)

[0209] In a spraying chamber, the leaves of appropriately aged sorghum cultivar Liangnuo No. 1 were sprayed with the compound of formula (I) prepared in Preparation Example 1 and the control agents (pyraclostrobin and difenoconazole). The sorghum cultivar was determined by spraying spores (1×10⁻⁶) 24 hours after application. 5 The detached leaves were inoculated with a suspension of (number of leaves per mL). The inoculated test plants were cultured in a greenhouse at 25°C and 90% relative humidity. When an appropriate level of disease appeared on the untreated control plants (7-14 days after application), the diseased area of ​​the leaves was counted, and the inhibition rate was calculated as follows: Inhibition rate = (Diseased area of ​​control group leaves - Diseased area of ​​treated group leaves) / Diseased area of ​​control group leaves.

[0210] The results showed that compound (I) had excellent control efficacy against ergot disease in sorghum.

[0211] Biological Example 21

[0212] In 2023, a wheat field trial was conducted at Baimahu Farm in Huai'an City, Jiangsu Province, China, to evaluate the efficacy of different compounds against wheat scab caused by Fusarium graminearum. Compounds of formula (I) were prepared as 10% SC in Formulation Example 1, 43% tebuconazole SC was purchased from Shandong Bainong Sida Biotechnology Co., Ltd., 30% prothioconazole OD was purchased from Anhui Jiuyi Agricultural Co., Ltd., and 45% imazalil EW was purchased from Zibo Hengsheng Pesticide Co., Ltd.

[0213] Apply the spray to the wheat ears at the early flowering stage using a boom sprayer with a flat, fan-shaped nozzle. The first application was on April 22nd; the second application was on April 28th. The coverage area was 180m² per region. 2 No duplicates allowed. Each treatment is 180m long. 2 = 0.27 mu. Weather conditions at the time of application: cloudy, high temperature 15℃, northeast wind force 4, gust force 7.

[0214] The disease occurred in early May, and its severity was assessed on May 18, 2023.

[0215] Survey Methodology:

[0216] Three sampling points were taken in each plot, and 200 ears of grain were surveyed at each point to investigate the severity and disease rate of each ear; the disease index and control effect were calculated.

[0217] Grading standards:

[0218] Level 0: No symptoms;

[0219] Grade 1: The length of the lesion is less than 25% of the length of the entire ear axis;

[0220] Grade 2: The length of the lesion accounts for 25-50% of the length of the entire ear axis;

[0221] Grade 3: The length of the lesion accounts for 50-75% of the length of the entire ear axis;

[0222] Grade 4: The length of the lesion is more than 75% of the length of the entire ear axis.

[0223] Disease index = ∑(number of diseased leaves at each level × relative level value) / (total number of ears surveyed × 9) × 100

[0224] Control efficacy (%) = (Disease index of blank control area - Disease index of treatment area) / Disease index of blank control area × 100

[0225] Test location:

[0226] Experimental site sowing crop variety resistance status Huai'an City Baimahu Farm November 28, 2022 wheat Zhenmai 3# Susceptible

[0227] Treatment List - Field Trials:

[0228]

[0229] Targets and crops in the experiment:

[0230] common name Latin name target Fusarium graminearum Fusarium graminearum crop wheat Triticum aestivum L.

[0231] Experimental layout:

[0232] Test methods field trials Experimental Design Then complete group Community area <![CDATA[180m 2 ]]>

[0233] Application details:

[0234] Application date April 22, 2023 and April 28, 2023 Application type boom sprayer with flat fan-shaped nozzle Application method Spraying wheat ears at the early flowering stage Slurry volume 450L / ha Application treatment Spraying with diluted pesticide

[0235] Efficacy assessment:

[0236]

[0237] in conclusion:

[0238] Compound (I) exhibits superior or comparable control efficacy against wheat scab, soybean anthracnose, potato late blight, wheat stripe rust, cucumber downy mildew, and cucumber powdery mildew compared to commercial standard agents. Compound (I) shows excellent activity against *Fusarium graminearum* on wheat at a ratio of 150 g ai / ha, with a control efficacy greater than 65%, comparable to tebuconazole at 180 g ai / ha and higher than prochloraz at 300 g ai / ha. For 30 days after application, compound (I) demonstrates sustained control of wheat scab, comparable to tebuconazole and prothioconazole, and longer than prochloraz.

[0239] Biological Example 22: The effect of the compound on inhibiting the DON toxin secreted by Fusarium graminearum on wheat.

[0240] Methods for determining DON (vomiting toxin):

[0241] The ELISA test kit used in this example was purchased from Savi Technology (catalog number: wsz0396):

[0242] One ELISA plate pre-coated with DON antigen (12 wells × 8 strips);

[0243] DON standard solutions: 5 bottles (1mL / bottle), with concentrations of 0ppb, 5ppb, 15ppb, 45ppb, and 135ppb respectively.

[0244] 1x Sample Diluent: 1 vial (50ml, for immediate use);

[0245] 10x concentrated detergent: 1 bottle (40mL);

[0246] Enzyme marker: 1 bottle (7 mL);

[0247] Antibody working solution: 1 bottle (7mL);

[0248] Colorimetric reagent: 1 bottle (15mL);

[0249] Stop solution: 1 bottle (8mL);

[0250] 2M H2SO4 (for adjusting sample pH): 1 bottle (2mL / bottle);

[0251] On May 22, 2023, wheat samples from each plot in Biological Example 21 were collected to determine the content of vomitoxin (DON) in the wheat grains in order to evaluate the effect of the compound on inhibiting DON toxin secreted by Fusarium graminearum on wheat.

[0252] Sampling method:

[0253] The control group was the plot in biological implementation series 21 that did not receive any chemical treatment, and the treatment group was the plot treated with 10% of compound (I) SC, 43% tebuconazole SC, 30% prothioconazole OD and 45% imazalil EW. Three samples were taken from each plot, 200 ears were investigated at each point, 10g of wheat grains were collected from each ear, and the grains were crushed to pass through a 20-mesh sieve.

[0254] Detection method:

[0255] Take 1g of the pulverized sample, add 20mL of water and mix well. Shake thoroughly for 3 minutes, centrifuge at 4000rpm at room temperature for 5 minutes, or filter using quantitative analysis filter paper. Take 0.5mL of the supernatant after centrifugation or the filtrate after filtration and add it to 0.5mL of the sample dilution solution for dilution and mix thoroughly. Adjust the pH of the diluted liquid to between 6 and 8, and take 50μL of the diluted liquid for testing.

[0256] Remove the required reagents and microplates and let them stand at room temperature for at least 30 minutes. All liquid reagents must be shaken well before use, and the washing working solution must also be brought to room temperature before use.

[0257] Number the wells corresponding to the samples and standards, and run two parallel wells for each sample and standard, recording the positions of the standard and sample wells. Add 50 μL of standard / sample to the corresponding well, then add 50 μL of label per well, followed by 50 μL of antibody working solution per well. Gently vortex to mix, cover with a cover film, and allow to cool to room temperature. Incubate in the dark for 15 min. Carefully remove the cover film and wash thoroughly with 250 μL of washing working solution, three times with 15-second intervals between washes. Pat dry with absorbent paper, add 100 μL of chromogenic solution per well, gently vortex to mix, cover with a cover film, and allow to cool to room temperature. Incubate in the dark for 5 min. Add 50 μL of stop solution per well, gently vortex to mix, and read the OD value of each well at 450 nm using a microplate reader (Synergy H1, Biotec).

[0258] Results analysis:

[0259] The average absorbance (B) of the measured standard solution or sample is divided by the absorbance (B0) of the first standard solution (0 standard) and then multiplied by 100% to obtain the percentage absorbance value.

[0260] Percent absorbance value (B / B0) × 100%

[0261] A standard curve is plotted with the base-10 logarithm of the standard concentration as the X-axis and the percentage absorbance as the Y-axis. The sample's percentage absorbance is substituted into the standard curve, and the corresponding value is read from the curve. This value is then raised to the power of 10 and multiplied by the dilution factor to determine the amount of vomitoxin in the sample.

[0262] The results showed that the compound of formula (I) could inhibit the DON toxin secreted by Fusarium graminearum on wheat and reduce the DON toxin content in wheat grains.

Claims

1. A method for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, said plant pathogenic microorganisms being selected from Fusarium graminearum, Botrytis cinerea, Phytophthora sacchariformis, Phytophthora capsici, Phytophthora virosa, Helicobacter spp., Colletotrichum gloeosporioides, Anthracnose spp., Pythium driezii, Pseudomonas spp., Pseudomonas spp., Pseudomonas cucumeroides, and Powdery mildew, said method comprising applying a compound of formula (I), or an agrochemically active salt thereof, to the plant, its location, or its propagation material:

2. The method according to claim 1, characterized in that, The agricultural chemically active salts are alkali addition salts of inorganic and organic bases.

3. The method according to claim 2, characterized in that, The agricultural chemically active salt is its potassium salt, sodium salt, ammonium salt, dimethylamine salt, or isopropylamine salt.

4. The method according to any one of claims 1 to 3, characterized in that, The useful plants are selected from wheat, barley, rice, sorghum, oats, tomatoes, strawberries, grapes, soybeans, peppers, potatoes, corn, apples, cucumbers, melons, okra, spinach, lettuce, asparagus, cabbage, carrots, onions, peppers, pears, peaches, walnuts, hawthorns, persimmons, dates, chestnuts, citrus fruits, lychees, zucchini, pumpkins, and cotton.

5. The method according to claim 1, characterized in that... The plant pathogenic microorganisms are selected from Fusarium graminearum, Botrytis cinerea, Phytophthora sacchariformis, Phytophthora capsici, Phytophthora pathogenica, Helicobacter spp., Colletotrichum gloeosporioides, Pythium driezii, Pseudomonas cocovenenans, Anthracnose spp., Pleurotus ostreatus, and Powdery mildew, and the plants are selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber, and cotton.

6. The method according to claim 5, characterized in that, The plant pathogenic microorganism is Fusarium graminearum, and the useful plant is wheat or rice.

7. The method according to any one of claims 1 to 3, characterized in that, The compound of formula (I) is prepared into a dosage form of one of emulsifiable concentrate, suspension concentrate, wettable powder, powder, granule, aqueous solution, poison bait, mother liquor and mother powder; the content of the compound in the drug is 1 to 99.9999% by weight; the effective dose concentration of the drug when used is 0.1 ug / ml to 100 ug / ml, and the application rate is 1 to 500 g / ha.

8. The method according to claim 7, characterized in that, The content of the drug is 10-80% by weight; the dosage of the drug is 1 to 500 g / ha.

9. The use of a compound of formula (I) or its agrochemically active salt for controlling or preventing infection of useful plants by plant pathogenic microorganisms, said plant pathogenic microorganisms being selected from Fusarium graminearum, Botrytis cinerea, Phytophthora sacchariformis, Phytophthora capsici, Phytophthora virosa, Helicobacter spp., Colletotrichum gloeosporioides, Anthracnose spp., Pythium driezii, Pseudomonas spp., Pseudomonas cucumeroides, and Powdery mildew.

10. The use according to claim 9, characterized in that, The agricultural chemically active salts are alkali addition salts of inorganic and organic bases.

11. The use according to claim 10, characterized in that, The agricultural chemically active salt is its potassium salt, sodium salt, ammonium salt, dimethylamine salt, or isopropylamine salt.

12. The use according to claim 9, wherein the plant is selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

13. The use according to claim 9, characterized in that, The plant pathogenic microorganism is Fusarium graminearum, and the useful plant is wheat or rice.

14. A method for growing useful plants, wherein the compound of method formula (I) or its agrochemically active salt is used to treat the propagation material for controlling or preventing infection of the useful plants by plant pathogenic microorganisms, said plant pathogenic microorganisms being selected from Fusarium graminearum, Botrytis cinerea, Phytophthora sacchariformis, Phytophthora capsici, Phytophthora virosa, Helicobacter spp., Colletotrichum gloeosporioides, Anthracnose spp., Pythium driezii, Pseudomonas stylosa, Pseudomonas copaiformis, and Powdery mildew.

15. The method as described in claim 14, characterized in that, The plants are selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

16. The method according to claim 15, characterized in that, The agricultural chemically active salts are alkali addition salts of inorganic and organic bases.

17. The method according to claim 15, characterized in that, The agricultural chemically active salt is its potassium salt, sodium salt, ammonium salt, dimethylamine salt, or isopropylamine salt.

18. The use of a compound of formula (I) or its agrochemically active salt in the preparation of a drug for controlling or preventing the infection of useful plants by plant pathogenic microorganisms, wherein the plant pathogenic microorganisms are selected from Fusarium graminearum, Botrytis cinerea, Phytophthora sacchariformis, Phytophthora capsici, Phytophthora virosa, Helicobacter spp., Colletotrichum gloeosporioides, Anthracnose spp., Pythium driezii, Pseudomonas stylosa, Pseudomonas copaiformis, and Powdery mildew.

19. The application as described in claim 18, characterized in that, The plants are selected from wheat, rice, sorghum, tomato, strawberry, soybean, pepper, potato, corn, apple, pear, cucumber and cotton.

20. The application as described in claim 18, characterized in that, The agricultural chemically active salts are alkali addition salts of inorganic and organic bases.

21. The application as described in claim 20, characterized in that, The agricultural chemically active salt is its potassium salt, sodium salt, ammonium salt, dimethylamine salt, or isopropylamine salt.

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