A bactericidal composition and use thereof

Abstract

The application belongs to the technical field of pesticides, and discloses a fungicidal composition and application thereof. The fungicidal composition comprises active ingredient A and active ingredient B. The active ingredient A is a compound of formula I. The active ingredient B is selected from any one of isoflucypram, fluoroimide and fluoroamide. The mass ratio of the active ingredient A to the active ingredient B is 1:35-32:1. The fungicidal composition has significant synergistic effect on various plant diseases in agriculture, is fast-acting, has long persistence, can delay the generation of pathogen resistance, and is suitable for comprehensive management of various plant diseases in agriculture.

Description

Technical Field

[0001] This invention relates to the field of pesticide sterilization technology, specifically to a sterilization composition and its uses. Background Technology

[0002] Isoflucypram is a succinate dehydrogenase inhibitor (SDHI) fungicide, CAS Registry Number: 1255734-28-1, chemical name: N-[(5-chloro-2-prop-2-ylphenyl)methyl]-N-cyclopropyl-3-(difluoromethyl)-5-fluoro-1-methylpyrazole-4-carboxamide. It can be used to control various diseases such as net blotch, leaf spot, leaf rust, stripe rust, and leaf blight, and has good control effects.

[0003] Fluopyram, CAS Registry No.: 1309859-39-9, Chemical Name: N-((3-chloro-5-(trifluoromethyl)pyridin-2-yl)methyl)-2,3,5,6-tetrafluoro-4-methoxybenzamide. Fluopyram mainly acts on the respiratory chain of fungal mitochondria, inhibiting the activity of succinate dehydrogenase, thereby affecting electron transport in mitochondria, hindering their respiration, and inhibiting fungal spore germination, germ tube elongation, hyphal growth, and sporogen formation.

[0004] Fluoramide, CAS Registry No.: 66332-96-5, Chemical Name: N-(3-prop-2-yloxyphenyl)-2-(trifluoromethyl)benzamide; Fluoramide acts as a succinate dehydrogenase inhibitor in the electron transport chain of respiration, inhibiting the synthesis of glutamate and aspartate. It is a systemic fungicide with both protective and curative effects, mainly used to control damping-off, leaf spot, white mold, rust, etc.

[0005] Succinate dehydrogenase inhibitors (SDHIs) act on succinate dehydrogenase in the respiratory chain of pathogenic fungi, blocking electron transport and interfering with fungal energy metabolism to exert their fungicidal effect. However, due to the single site of action of SDHIs and their widespread use in agricultural production, resistance problems are becoming increasingly prominent. Resistant strains have emerged in the field, leading to reduced efficacy and increased application rates. To delay the occurrence and development of resistance and improve the control effect of diseases in the field, the inventors selected a compound of formula I with a different mechanism of action than SDHIs and rationally compounded it with SDHIs. This improved the control effect, broadened the control spectrum, and delayed the development of pathogen resistance. Summary of the Invention

[0006] Based on the above, the purpose of this invention is to provide a bactericidal composition that is effective against a variety of plant pathogens, significantly enhances efficacy, and is superior to single agents in terms of both rapid and sustained effects. It also reduces the amount of pesticides used in agricultural production, slows the development of pesticide resistance in pathogens, reduces pesticide residues in agricultural products, and is suitable for the integrated management of various diseases in agriculture.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: a bactericidal composition, wherein the bactericidal composition comprises active ingredient A and active ingredient B, wherein active ingredient A is a compound of formula I: The active ingredient B is selected from any one of isoflucypram, fluopyram, and fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:35 to 32:1, or any value within the above range.

[0008] Furthermore, the active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:32 to 20:1, or any value within the above range.

[0009] The active ingredient B is fluopyram, and the mass ratio of the active ingredient A to the active ingredient B is 1:35 to 20:1, or any value within the above range.

[0010] The active ingredient B is fluoroamide, and the mass ratio of the active ingredient A to the active ingredient B is 1:20 to 30:1, or any value within the above range.

[0011] Furthermore, the active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:25 to 15:1, or any value within the above range.

[0012] The active ingredient B is fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:22 to 15:1, or any value within the above range.

[0013] The active ingredient B is fluoroamide, and the mass ratio of active ingredient A to active ingredient B is 1:15 to 20:1, or any value within the above range.

[0014] Furthermore, the active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:18 to 10:1, or any value within the above range.

[0015] The active ingredient B is fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:16 to 10:1, or any value within the above range.

[0016] The active ingredient B is a fluoroamide, and the mass ratio of the active ingredient A to the active ingredient B is 1:5 to 18:1, or any value within the above range.

[0017] Furthermore, based on a total weight of 100 wt%, the total weight of active ingredient A and active ingredient B accounts for 1% to 80% of the total weight of the bactericidal composition.

[0018] Furthermore, the bactericidal composition includes other auxiliary components in addition to the active ingredients, and the auxiliary components are selected from one or more of the following: wetting agents, dispersants, emulsifiers, thickeners, disintegrants, antifreeze agents, defoamers, solvents, preservatives, stabilizers, warning colors, film-forming agents, synergists, and carriers.

[0019] Furthermore, the bactericidal composition can be prepared into any agriculturally acceptable formulation, which may be a solid or liquid formulation.

[0020] Furthermore, the bactericidal composition can be prepared into any agriculturally acceptable formulation, which may be a solid formulation, a liquid formulation, and / or a seed treatment formulation.

[0021] Furthermore, the solid dosage form is a direct-use solid dosage form, a dispersible solid dosage form, or a soluble solid dosage form;

[0022] Furthermore, the directly usable solid dosage form is a powder, granule, ball, tablet, or strip;

[0023] The dispersible solid dosage form is a wettable powder, an oil-dispersible powder, an emulsion powder, a water-dispersible granule, an emulsion granule, or a water-dispersible tablet;

[0024] The soluble solid dosage form is a soluble powder, soluble tablet, or soluble granule;

[0025] Furthermore, the liquid formulation is a solution formulation, a dispersed liquid formulation, an emulsion formulation, a suspension formulation, or a multiphase formulation;

[0026] Furthermore, the solution formulation is a soluble agent, a colloid, an oil, or a film-spreading oil;

[0027] The dispersed liquid formulation is an emulsifiable concentrate, latex, dispersible liquid, or ointment;

[0028] The emulsion formulation is an aqueous emulsion, an oil emulsion, a microemulsion, or a lipid formulation;

[0029] The suspension formulation is a suspension agent, microcapsule suspension agent, oil suspension agent, or dispersible oil suspension agent;

[0030] The multiphase formulation is a suspension emulsion, a microcapsule suspension-suspension, a microcapsule suspension-water emulsion, or a microcapsule suspension-suspension emulsion;

[0031] The solid formulation is a water-dispersible granule and / or a wettable powder, and the liquid formulation is a suspension.

[0032] The present invention also discloses the use of the bactericidal composition described above in the prevention or control of crop diseases.

[0033] Furthermore, the crops are selected from food crops, cash crops, fruits and vegetables, and / or lawn and garden crops;

[0034] The diseases mentioned are wheat sheath blight, wheat rust, soybean rust, rice bakanae disease, rice sheath blight, rice seedling blight, cucumber seedling blight, potato late blight, cucumber downy mildew, grape downy mildew, wheat powdery mildew, cucumber powdery mildew, and cucumber anthracnose.

[0035] Furthermore, the diseases mentioned are rice sheath blight, wheat sheath blight, or cucumber downy mildew.

[0036] This invention also discloses a method for preventing and controlling crop pathogens, wherein the fungicidal composition and / or its preparations are applied in an effective and substantially non-phytotoxic amount to plants, plant propagation materials and subsequently growing plant organs, cultivation media, materials or spaces by means of seed treatment, foliar application, stem application, soaking, dripping, watering, spraying, misting, dusting, dispersing or fumigation.

[0037] Compared with the prior art, the advantages of the present invention are as follows:

[0038] 1) The bactericidal composition of the present invention has a significant synergistic effect by rationally combining compounds with different mechanisms of action, which improves the activity against pathogens, enhances the field control effect, and helps to delay the development of drug resistance in pathogens;

[0039] 2) The bactericidal composition of the present invention reduces the amount of pesticides used in agricultural production, reduces pesticide residues in agricultural products, is environmentally friendly, and is conducive to the comprehensive management of agricultural diseases. Detailed Implementation

[0040] To better understand the essence of the present invention, the following embodiments further illustrate the content of the present invention, but these should not be regarded as limitations on the present invention. The content mentioned in the embodiments is not a limitation of the present invention, and the selection of material formulations can be adapted to local conditions without having a substantial impact on the results.

[0041] Formulation preparation example:

[0042] Preparation Example 1: 26% Formula I compound·isoflucypram suspension (1:12)

[0043] By weight percentage, 2% Formula I compound, 24% isoflucypram, 2% sodium dodecyl sulfate, 1% ethylene glycol oxyethylene polyoxypropylene ether, 4% tristyrene-phenol polyoxyethylene ether phosphate, 0.25% xanthan gum, 1% magnesium aluminum silicate, 5% ethylene glycol, 1% sodium sorbate, 0.5% silicone oil, and deionized water to make up the balance.

[0044] Preparation method: According to the formula ratio, the active ingredients, surfactants and other functional additives are placed in the reaction vessel in sequence, water is added and mixed evenly, and then subjected to high-speed shearing, wet sand milling and finally homogenization filtration to obtain the suspension product.

[0045] Preparation Example 2: 24% formula I compound · isoflucypram water-dispersible granules (1:5)

[0046] By weight percentage, 4% Formula I compound, 20% isoflucypram, 5% naphthalene sulfonate formaldehyde condensate, 10% sodium lignosulfonate, 4% bleaching powder BX, 5% white sugar, and kaolin to make up the balance.

[0047] Preparation method: According to the formulation ratio in the example, the active ingredient is added to the carrier, and surfactants and other functional additives are added thereto. After mixing, the mixture is pulverized by air jet and 10-25% water is added. Then, the mixture is kneaded, granulated, dried and sieved to obtain a water-dispersible granule product; or the pulverized powder is sprayed with water, granulated and dried in a fluidized bed granulator, and then sieved to obtain the product.

[0048] Preparation Example 3: 33% Formula I compound·isoflucypram wettable powder (10:1)

[0049] By weight percentage, 30% of Formula I compound, 3% isoflucypram, 10% succinate sulfonate, 3% calcium lignosulfonate, 2% sodium lignosulfonate, 3% sodium dodecyl sulfate, 10% silica, and kaolin to make up the balance.

[0050] Preparation method: According to the formula ratio, the active ingredients, dispersant, wetting agent and filler are mixed and stirred evenly in a stirring tank. The mixture is then pulverized and mixed evenly multiple times by an air jet mill to prepare the wettable powder of the composition of the present invention.

[0051] Preparation Example 4: 20% Formula I compound·fluopyram suspension (2:3)

[0052] By weight percentage: 8% Formula I compound, 12% fluopyram, 2% sorbitan oleate polyoxyethylene ether, 3% arylphenol polyoxyethylene ether phosphate, 2% sodium lignosulfonate, 0.25% xanthan gum, 1.5% magnesium aluminum silicate, 5% propylene glycol, 0.5% silicone oil, 0.01% isothiazolinone, deionized water to make up the balance;

[0053] Preparation method: Same as in preparation example 1.

[0054] Preparation Example 5: 36% Formula I compound·fluopyram water-dispersible granules (1:8)

[0055] By weight percentage, 4% Formula I compound, 32% fluopyram, 1.5% sodium dodecyl sulfate, 5% sodium polycarboxylate, 10% naphthalene sulfonate formaldehyde condensate, 12% ammonium sulfate, and starch to make up the balance;

[0056] Preparation method: Same as in preparation example 2.

[0057] Preparation Example 6: 30% Formula I compound·flufenican wettable powder (5:1)

[0058] By weight percentage, 25% of Formula I compound, 5% fluopyram, 3% sodium polycarboxylate, 5% naphthalene sulfonate formaldehyde condensate, 5% dispersant NNO, 2% sodium dodecyl sulfate, and kaolin to make up the balance;

[0059] Preparation method: Same as in preparation example 3.

[0060] Preparation Example 7: 16% Formula I compound·fluoroamide suspension (1:15)

[0061] By weight percentage, 1% Formula I compound, 15% fluoroamide, 2% alkyl aryl polyoxyethylene ether polyoxypropylene ether, 1% naphthalene sulfonate formaldehyde condensate, 5% arylphenol polyoxyethylene ether phosphate, 0.2% xanthan gum, 1% magnesium aluminum silicate, 5% ethylene glycol, 0.1% sodium benzoate, 0.25% silicone oil, and deionized water to make up the balance;

[0062] Preparation method: Same as in preparation example 1.

[0063] Preparation Example 8: 39% Formula I compound·fluoroamide water-dispersible granules (12:1)

[0064] By weight percentage, 36% of Formula I compound, 3% fluoroamide, 8% lignin sulfonate, 5% naphthalene sulfonate formaldehyde condensate, 2% fatty alcohol polyoxyethylene ether sulfate, 5% silica, 30% starch, and kaolin to make up the balance.

[0065] Preparation method: Same as in preparation example 2.

[0066] Preparation Example 9: 28% Formula I compound·fluoroamide wettable powder (3:1)

[0067] By weight percentage, 21% of Formula I compound, 7% fluoroamide, 2% sodium lignosulfonate, 5% fatty alcohol polyoxyethylene ether sulfate, 3% stretching powder BX, 5% silica, and kaolin to make up the balance.

[0068] Preparation method: Same as in preparation example 3.

[0069] Indoor activity assay:

[0070] Example 1: Indoor activity assay of compound I combined with isoflucypram against wheat sheath blight

[0071] Test basis: The test refers to NY / T 1156.2-2006, the agricultural industry standard of the People's Republic of China, "Guidelines for Indoor Bioassay Tests of Pesticides - Fungicides Part 2: Test on Inhibition of Mycelial Growth of Pathogenic Fungi - Plate Method".

[0072] Test strain: Rhizoctonia cerealis.

[0073] Test material preparation: Transfer the pathogen stored in the refrigerator to PDA medium plates and incubate at (26±2)℃ for 2 days before use.

[0074] Test reagents: Formula I compound technical grade, isoflucypram technical grade.

[0075] Preparation of pharmaceutical stock solution: Dissolve the above raw materials separately with a suitable solvent to prepare a high-concentration stock solution, then dilute with 0.1% Tween 80 aqueous solution to prepare single-agent stock solutions. Design different ratios according to the purpose of mixing and the activity of the pharmaceuticals. Prepare each single agent and each group of mixed solutions to the required series of mass concentrations.

[0076] Experimental replication: Four petri dishes were used for each concentration of the test reagent, with one petri dish for each replicate, for a total of four replicates. A 0.1% Tween 80 aqueous solution without the reagent was used as a blank control.

[0077] Drug processing: Under aseptic conditions, use a pipette to add 5 mL of drug solution of different concentrations to pre-calibrated sterile Erlenmeyer flasks. Then, add the melted and cooled culture medium to the Erlenmeyer flasks, shake well, and pour the mixture into 4 petri dishes in equal amounts to prepare drug-containing PDA plates of the corresponding concentrations.

[0078] Inoculation: Under aseptic conditions, cut off a mycelial cake from the edge of the pre-cultured Rhizoctonia graminearum, inoculate the mycelial cake into the center of the drug-containing plate using an inoculator, cover with the cap, and place in a constant temperature incubator at 25°C for dark incubation.

[0079] Data collection: An experimental investigation was conducted based on the colony growth to 2 / 3 to 4 / 5 of the petri dish diameter in the control treatment. The colony diameter (cm) was measured using a ruler, and the diameter of each colony was measured once using the cross-sectional method, and the average value was taken.

[0080] Data statistics and analysis: Based on the survey results, the inhibition rate of mycelial growth of the tested target bacteria by each treatment concentration was calculated, in percentage (%). The calculation results were retained to two decimal places.

[0081] D = D1 - D2

[0082] In the formula:

[0083] D – Colony growth diameter;

[0084] D1—colony diameter;

[0085] D2 – Diameter of the mushroom cake.

[0086]

[0087] In the formula:

[0088] I – Mycelial growth inhibition rate;

[0089] D0—Correlation diameter of the blank control group;

[0090] D T — Diameter of colonies grown after chemical treatment.

[0091] Analyze using a statistical analysis system to determine the virulence regression line and EC. 50 The value is used to evaluate the activity of the test reagent on the biological sample.

[0092] Sun Yunpei's method: The synergistic effect of drug mixtures is evaluated based on the co-toxicity coefficient (CTC). A CTC ≥ 120 indicates a synergistic effect; a CTC ≤ 80 indicates an antagonistic effect; and a CTC < 120 indicates an additive effect.

[0093] The co-toxicity coefficient (CTC value) of the mixture is calculated using the following formula:

[0094]

[0095] In the formula:

[0096] ATI – Actual Measured Toxicity Index of Mixtures;

[0097] S—EC of standard reagent 50 The unit is milligrams per liter (mg / L);

[0098] M – EC of the mixture 50The unit is milligrams per liter (mg / L).

[0099] TTI = TI A ×P A +TI B ×P B

[0100] In the formula:

[0101] TTI – Theoretical Toxicity Index of Mixtures;

[0102] TI A —A. Toxicity index of drug A;

[0103] P A —Percentage content of drug A in the mixture, expressed as percentage (%);

[0104] TI B —Toxicity index of drug B;

[0105] P B —Percentage content of agent B in the mixture, expressed as percentage (%).

[0106]

[0107] In the formula:

[0108] CTC – Cotoxicity Coefficient;

[0109] ATI – Actual Measured Toxicity Index of Mixtures;

[0110] TTI – Theoretical Toxicity Index of Mixtures.

[0111] The test results are shown in the table below:

[0112] Table 1 shows the results of the indoor activity assay of compound I in combination with isoflucypram against wheat sheath blight.

[0113]

[0114] As shown in Table 1, the test compound I and isoflucypram have good control effects against wheat sheath blight pathogens, and their EC50 values ​​are [not specified]. 50The concentrations were 0.2196 mg / L and 1.6540 mg / L, respectively. The co-toxicity coefficients of compound I with isoflucypram were greater than 120 in the mass ratio range of 1:32 to 20:1, indicating a synergistic effect against wheat sheath blight. The co-toxicity coefficients were greater than 130 in the mass ratio range of 1:25 to 15:1, showing a significant synergistic effect. The co-toxicity coefficients were greater than 150 in the mass ratio range of 1:18 to 5:1, also showing a significant synergistic effect against wheat sheath blight. The highest co-toxicity coefficient and the most significant synergistic effect were observed when compound I was combined with isoflucypram at a mass ratio of 1:5.

[0115] Example 2: Indoor activity test of compound I combined with fluopyram against cucumber downy mildew.

[0116] Experimental basis: The experiment was conducted in accordance with NY / T 1156.7-2006 "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 7: Pot Test for Control of Downy Mildew in Cucumber".

[0117] Experimental target: Cucumber downy mildew (Pseudoperonospora cubensis).

[0118] Experimental instruments and equipment: electronic balance, spray equipment, artificial climate chamber, biological incubator, petri dishes, pipettes, etc.

[0119] Test material preparation: Select potted cucumber varieties susceptible to disease (Xintai Mici), and prepare seedlings when they have grown to 4 to 6 true leaves.

[0120] Test reagents: Formula I compound technical grade, fluopyram technical grade.

[0121] Experimental steps:

[0122] Select infected cucumber leaves, wash off the sporangia of downy mildew from the underside of the leaves with distilled water at 4℃, prepare a suspension, and store at 4℃ for later use. Dissolve the original drug in a suitable solvent and then dilute with a 0.1% Tween 80 aqueous solution. Based on the drug activity, set up 5 series of mass concentrations. Spray the fresh sporangia suspension onto the underside of the leaves, with 5 pots per treatment, 2 plants per pot, and 4 replicates per treatment. Apply the drug 24 hours after inoculation. According to the experimental design, spray the drug evenly on both sides of the leaves until completely wet. A control treatment without the drug was set up as a blank control. After application, the plants were placed under conditions of photoperiod of light:dark = 12h:12h, temperature of 17–22℃, and relative humidity of 90%–95%. Based on the disease incidence of the blank control, the inoculated leaves were graded and investigated. 30 leaves were investigated for each treatment.

[0123] The following grading method was used to conduct a grading survey and record the results:

[0124] Level 0: No disease;

[0125] Grade 1: The area of ​​lesions accounts for less than 5% of the total leaf area;

[0126] Grade 3: The lesion area accounts for 6% to 10% of the total leaf area;

[0127] Level 5: The lesion area accounts for 11% to 25% of the total leaf area;

[0128] Level 7: The lesion area accounts for 26% to 50% of the total leaf area;

[0129] Level 9: The lesion area accounts for more than 50% of the total leaf area;

[0130] Data calculation:

[0131] Based on the data survey, the disease index and prevention and control effects of each treatment were calculated.

[0132] The disease index is calculated using the following formula:

[0133]

[0134] The prevention and control effect is calculated using the following formula:

[0135]

[0136] Statistical analysis:

[0137] Analyze using a statistical analysis system to determine EC. 50 The value is used to evaluate the activity of the test reagent on the biological sample.

[0138] The co-toxicity coefficient (CTC value) of the mixture is calculated using the following formula:

[0139]

[0140] In the formula:

[0141] ATI – Actual Measured Toxicity Index of Mixtures;

[0142] S—EC of standard reagent 50 The unit is milligrams per liter (mg / L);

[0143] M – EC of the mixture 50 The unit is milligrams per liter (mg / L).

[0144] TTI = TI A ×P A +TI B ×P B

[0145] In the formula:

[0146] TTI – Theoretical Toxicity Index of Mixtures;

[0147] TI A —A. Toxicity index of drug A;

[0148] P A —Percentage content of drug A in the mixture, expressed as percentage (%);

[0149] TI B —Toxicity index of drug B;

[0150] P B —Percentage content of agent B in the mixture, expressed as percentage (%).

[0151]

[0152] In the formula:

[0153] CTC – Cotoxicity Coefficient;

[0154] ATI – Actual Measured Toxicity Index of Mixtures;

[0155] TTI – Theoretical Toxicity Index of Mixtures.

[0156] The co-toxicity coefficient of the compound is ≥120, which shows a synergistic effect; CTC≤80 shows an antagonistic effect; and 80<CTC<120 shows an additive effect.

[0157] The results of the indoor tests are shown in the table below:

[0158] Table 2 shows the results of indoor activity assays of compound I combined with fluopyram to treat cucumber downy mildew.

[0159]

[0160] As shown in Table 2, the test compound I and fluopyram have good therapeutic effects on cucumber downy mildew, with EC50... 50 The concentrations were 6.2910 mg / L and 15.5076 mg / L, respectively. When the mass ratio of Compound I to fluopyram was between 1:35 and 20:1, the co-toxicity coefficient was greater than 120, indicating a synergistic effect against cucumber downy mildew. When the mass ratio of Compound I to fluopyram was between 1:22 and 15:1, the co-toxicity coefficient was greater than 130, showing a significant synergistic effect. When the mass ratio of Compound I to fluopyram was between 1:16 and 10:1, the co-toxicity coefficient was greater than 150, showing a significant synergistic effect against cucumber downy mildew. Among these, the co-toxicity coefficient was the highest and the synergistic effect was most significant when Compound I was combined with fluopyram at a mass ratio of 2:3.

[0161] Example 3: Indoor activity assay of compound I combined with fluoroamide against rice sheath blight.

[0162] Test basis: The test refers to NY / T 1156.2-2006, the agricultural industry standard of the People's Republic of China, "Guidelines for Indoor Bioassay Tests of Pesticides - Fungicides Part 2: Test on Inhibition of Mycelial Growth of Pathogenic Fungi - Plate Method".

[0163] Experimental target: Rhizoctonia solani.

[0164] Instruments and equipment: moist heat sterilizer, ultra-clean workbench, constant temperature light incubator, electric heating drum windproof drying oven, 0.01% electronic balance, pipette, alcohol lamp, small beakers, volumetric flasks, Erlenmeyer flasks, petri dishes, hole punch, inoculator, ruler, etc.

[0165] Culture conditions for the test target: Rhizoctonia solani stored in an indoor refrigerator was transferred to potato dextrose agar medium and incubated in the dark at 26°C for 4 days to activate it for later use.

[0166] Test reagents: Compound of Formula I and fluoroamide technical.

[0167] Preparation of pharmaceutical stock solution: Dissolve the above raw materials separately with a suitable solvent to prepare a high-concentration stock solution, then dilute with 0.1% Tween 80 aqueous solution to prepare single-agent stock solutions. Design different ratios according to the purpose of mixing and the activity of the pharmaceuticals. Prepare each single agent and each group of mixed solutions to the required series of mass concentrations.

[0168] Experimental replication: Four petri dishes were used for each concentration of the test reagent, with one petri dish for each replicate, for a total of four replicates. A 0.1% Tween 80 aqueous solution without the reagent was used as a blank control.

[0169] Drug treatment: Under aseptic conditions, according to the test treatment, pre-melted and sterilized PDA culture medium was quantitatively added to a sterile conical flask. 10 mL of each prepared treatment solution was quantitatively taken from low concentration to high concentration and added to the conical flasks respectively. The solutions were shaken thoroughly and then poured into four 9 cm diameter petri dishes to prepare drug-containing plates of the corresponding concentrations.

[0170] Inoculation: Cut off a mycelial cake from the edge of the pre-cultured Rhizoctonia solani colony, inoculate the mycelial cake into the center of the drug-containing plate using an inoculator, cover with the cap, and place in a constant temperature and light incubator at (25±1)℃ for dark incubation.

[0171] Data collection: The growth of pathogenic fungal hyphae was investigated based on the growth of hyphae in the blank control culture dishes. The diameter of the colonies was measured in centimeters (cm) using a ruler. The diameter of each colony was measured once using the cross-sectional method, and the average value was taken. The original data of all replicates for each treatment were recorded.

[0172] Data statistics and analysis: Based on the survey results, the inhibition rate of mycelial growth of the tested target bacteria by each treatment concentration was calculated, in percentage (%). The calculation results were retained to two decimal places.

[0173] D = D1 - D2

[0174] In the formula:

[0175] D – Colony growth diameter;

[0176] D1—colony diameter;

[0177] D2 – Diameter of the mushroom cake.

[0178]

[0179] In the formula:

[0180] I – Mycelial growth inhibition rate;

[0181] D0—Correlation diameter of the blank control group;

[0182] D T — Diameter of colonies grown after chemical treatment.

[0183] The data were processed using probability value analysis. A statistical analysis system was used to determine the toxicity regression line and EC50. 50 Values ​​and correlation coefficient R 2 To evaluate the activity of the test reagent on biological materials.

[0184] Sun Yunpei's method: The synergistic effect of drug mixtures is evaluated based on the co-toxicity coefficient (CTC). A CTC ≥ 120 indicates a synergistic effect; a CTC ≤ 80 indicates an antagonistic effect; and a CTC < 120 indicates an additive effect.

[0185] The co-toxicity coefficient (CTC value) of the mixture is calculated using the following formula:

[0186]

[0187] In the formula:

[0188] ATI – Actual Measured Toxicity Index of Mixtures;

[0189] S—EC of standard reagent 50 The unit is milligrams per liter (mg / L);

[0190] M – EC of the mixture 50 The unit is milligrams per liter (mg / L).

[0191] TTI = TI A ×P A +TI B ×P B

[0192] In the formula:

[0193] TTI – Theoretical Toxicity Index of Mixtures;

[0194] TI A —A. Toxicity index of drug A;

[0195] P A —Percentage content of drug A in the mixture, expressed as percentage (%);

[0196] TI B —Toxicity index of drug B;

[0197] P B —Percentage content of agent B in the mixture, expressed as percentage (%).

[0198]

[0199] In the formula:

[0200] CTC – Cotoxicity Coefficient;

[0201] ATI – Actual Measured Toxicity Index of Mixtures;

[0202] TTI – Theoretical Toxicity Index of Mixtures.

[0203] The test results are shown in the table below:

[0204] Table 3 shows the results of the indoor activity assay of compound I combined with fluoroamide against rice sheath blight.

[0205]

[0206] Table 3 shows that the tested compound I and fluopyram have good control effects on rice sheath blight pathogens, and their EC50 values ​​are [not specified]. 50The concentrations were 0.5567 mg / L and 0.2694 mg / L, respectively. When the mass ratio of Compound I to flumethamide was between 1:20 and 30:1, the co-toxicity coefficient was greater than 120, indicating a synergistic effect against rice sheath blight. When the mass ratio of Compound I to flumethamide was between 1:15 and 20:1, the co-toxicity coefficient was greater than 130, showing a significant synergistic effect against rice sheath blight. When the mass ratio of Compound I to flumethamide was between 1:5 and 18:1, the co-toxicity coefficient was greater than 150, showing a significant synergistic effect against rice sheath blight. Among these, the co-toxicity coefficient was the highest and the synergistic effect was most significant when Compound I and flumethamide were combined at a mass ratio of 3:1.

[0207] Field efficacy trials:

[0208] Example 4: Field efficacy trial for controlling wheat sheath blight

[0209] Experimental site: Wheat field in Sunjiashangou Village, Yanzhuang Town, Ju County, Shandong Province. The soil in the experimental site is alluvial brown soil with medium to high fertility. Wheat sheath blight has occurred widely and evenly over the years.

[0210] Experimental target: Wheat sheath blight.

[0211] Experimental crop: Wheat (Luyuan 502).

[0212] Experimental Design: The experiment consisted of 8 treatments, with the dosage for each treatment shown in the table below. Water spraying served as the control (CK). The experiment was repeated 4 times, with a total of 32 plots arranged in a randomized block design. Each plot area was 72 m². 2 .

[0213] Application time: The experiment was conducted by applying the pesticide once at the end of the wheat jointing stage. The entire wheat plant was manually sprayed using a Gongnong-16 backpack sprayer.

[0214] Experimental survey: Five random points were selected in each plot, with 50 plants at each point. Disease incidence was investigated 25 days after application of the pesticide. Disease incidence was investigated according to the following grading standards, and the disease index and control effect were calculated.

[0215] Grading standards:

[0216] Level 0, no disease;

[0217] Grade 1: Leaf sheaths are infected, but stems are not.

[0218] Level 3: The leaf sheath is infected and the disease invades the stem, but the lesions on the stem cover less than half of the stem.

[0219] Level 5: Stem lesions encircle more than half of the stem, but the stem does not collapse or break.

[0220] Level 7: Dead, lodged, withered white ears.

[0221] Calculation method:

[0222]

[0223] The results of the field efficacy trials are shown below:

[0224] Table 4 Results of field efficacy trials for controlling wheat sheath blight

[0225] serial number Drug Name <![CDATA[Dosage of active ingredient (g / hm 2 )]]> Disease index Prevention and control efficacy (%) 1 Preparation Example 9: 28% Formula I compound·fluoroamide wettable powder (3:1) 185 1.29 95.16 2 Preparation Example 8: 29% Formula I compound·fluoroamide water-dispersible granules (12:1) 185 3.34 87.41 3 Preparation Example 2: 24% formula I compound · isoflucypram water-dispersible granules (1:5) 185 1.81 93.17 4 Preparation Example 1: 26% Formula I compound·isoflucypram suspension (1:12) 185 2.99 88.76 5 25% isoflucypram wettable powder 225 9.06 65.90 6 20% Fluoramide Water Dispersible Granules 225 7.46 71.92 7 20% Formula I compound suspension 225 6.17 76.76 8 Blank control / 26.56 /

[0226] 25 days after application, all seven agents showed some control effect against wheat sheath blight. Preparation Example 9: 28% Formula I compound·flumethamide wettable powder (3:1) had the highest control efficacy, reaching 95.16%; followed by Preparation Example 2: 24% Formula I compound·isoflucypram water-dispersible granules (1:5) treatment group, with a control efficacy of 93.17%. Compared with the blank control, the compound preparations showed significant control effect against wheat sheath blight.

[0227] Example 5: Field efficacy trial for controlling rice sheath blight

[0228] Experimental basis: The experiment was conducted in accordance with GB / T 17980.20-2000 "Field Efficacy Test Guidelines for Pesticides (I) Control of Rice Sheath Blight with Fungicides".

[0229] Experimental target: Rice sheath blight (Thanatephorus cucumeris).

[0230] Experimental crop: Rice (Huai Dao No. 5).

[0231] The experiment was conducted in Dali Village, Yanlong Street, Yandu District, Yancheng City, Jiangsu Province. The experimental field was a rice-wheat rotation field, with wheat as the previous crop. The soil texture of the experimental field was clay, with good soil fertility. The management practices within the experimental area were consistent and in line with local agricultural production practices.

[0232] Test reagents: The test reagents and dosages are shown in the table below.

[0233] Experimental design: Cells were randomly arranged, with each cell having an area of ​​20m². 2 Each treatment was repeated 4 times.

[0234] Experimental Implementation: Two applications of pesticide were applied on August 5th and August 26th, 2024 (late tillering stage of rice) and early booting stage of rice, respectively. A 3WBD-18 backpack electric sprayer was used for uniform spraying of the entire plant. The sprayer's operating pressure was 0.15–0.40 MPa, and the spray width was 4–5 m, covering an area of ​​667 m². 2 The dosage is 30 kg of water. Irrigate the field with water to a depth of 5-7 cm before application and keep the water level for 6 days.

[0235] Investigation Methods: After each application of pesticides, visual inspection was conducted to periodically observe the growth and development of rice in each plot to evaluate the safety of each pesticide on rice growth. An investigation was carried out 8 days after the last application. Based on the severity of damage to the rice leaf sheath and leaves, samples were taken at five points diagonally across each plot, with five adjacent clumps sampled at each point, for a total of 25 clumps. The total number of plants, the number of diseased plants, and the disease severity were recorded.

[0236] Disease severity grading criteria:

[0237] Grade 0: The entire plant is disease-free;

[0238] Grade 1: Disease occurs on the fourth leaf and all leaf sheaths and leaves below it (with the sword leaf as the first leaf);

[0239] Grade 3: Disease occurs on the third leaf and all leaf sheaths and leaves below it;

[0240] Level 5: Disease occurs on the second leaf and all leaf sheaths and leaves below it;

[0241] Level 7: Disease occurs on the sword-shaped leaf blade and all leaf sheaths and leaves below it;

[0242] Level 9: The entire plant is infected and dies prematurely.

[0243] Methods for calculating drug efficacy:

[0244]

[0245] The results of field efficacy trials are shown in the table below:

[0246] Table 5 Results of field efficacy trials for controlling rice sheath blight.

[0247]

[0248] As shown in Table 5, the control efficacy of the compound of Formula I in different mixtures with fluoxetine or isoflucypram against rice sheath blight was superior to that of the control agent. Eight days after the last application, the fungicidal composition of the present invention achieved a maximum control efficacy of 92.86%.

[0249] Example 6: Field efficacy test of pesticide for controlling cucumber downy mildew

[0250] This experiment was conducted in accordance with GB / T 17980.26-2000 "Guidelines for Field Efficacy Tests of Pesticides (I) Control of Downy Mildew in Cucumbers with Fungicides".

[0251] Experimental Site: The experimental site is located in a greenhouse in Henan Village, Hanting District, Weifang City. The greenhouse is a solar greenhouse with relatively flat terrain and sandy loam soil. Since cucumbers are grown in this greenhouse year-round, cucumber downy mildew occurs frequently.

[0252] Experimental target: Cucumber downy mildew (Pseudoperonospora cubensis).

[0253] Experimental crop: Cucumber (Xintai Dense Thorn).

[0254] Experimental Design: This experiment consisted of 6 treatments, including 3 treatments with compound agents, 2 single-agent controls, and 1 water control. Each treatment was replicated 4 times, and each plot was 15 m². 2 Isolation rows are set up on both sides and between the small areas, and the cultivation conditions and field cultivation management are consistent in each small area.

[0255] Experimental Methods: The experiment involved two applications of pesticide. The first application was initiated when sporadic cases of cucumber downy mildew were observed in the greenhouse, followed by a second application 7 days later. A HeMei HM-16A backpack electric sprayer (manufactured by Zhongshan HeMei Electric Appliance Co., Ltd., operating pressure 0.2–0.4 MPa) was used to evenly spray the cucumber plants with water, using 45 kg of water per acre. Spraying was continued until both sides of the leaves were evenly covered and moistened. The cucumbers were in the initial flowering stage to peak fruit set stage during the experiment.

[0256] Experimental investigation: The control effect was investigated 7 days after the first application and 10 days after the second application, for a total of 2 investigations. During the investigation, a 5-point sampling method was used in each plot, with 2 plants sampled at each point, for a total of 10 plants. All leaves of each plant were investigated. The percentage of diseased area per leaf was used to classify the treatments, and the number of diseased leaves for each treatment was recorded. The disease index and control effect were calculated.

[0257] Throughout the experiment, the safety of each pesticide treatment on cucumbers was observed periodically, and no adverse effects of the tested pesticides on cucumbers were found.

[0258] Grading method:

[0259] Grade 0, no lesions;

[0260] Grade 1: Lesions cover less than 5% of the total leaf area;

[0261] Grade 3, with lesions covering 6% to 10% of the total leaf area;

[0262] Level 5, with lesions covering 11% to 25% of the total leaf area;

[0263] Level 7, with lesions covering 26% to 50% of the total leaf area;

[0264] Level 9, with lesions covering more than 51% of the total leaf area.

[0265] The disease index and prevention efficacy are calculated using the following formula:

[0266]

[0267] The results of the field efficacy trials are shown in the table below:

[0268] Table 6 Results of field efficacy trials for controlling cucumber downy mildew.

[0269]

[0270] Field efficacy trials (Table 6) showed that, 10 days after the second application, the control efficacy of the following preparations—Example 6: 30% Formula I compound·fluopyram wettable powder (5:1), Example 4: 20% Formula I compound·fluopyram suspension (2:3), and Example 5: 36% Formula I compound·fluopyram water-dispersible granules (1:8)—was 87%–92%. Therefore, the rational combination of Formula I compound and fluopyram has a good control effect on cucumber downy mildew and can effectively slow the spread of the disease.

[0271] Indoor toxicity tests and field efficacy tests show that the fungicidal composition of the present invention exhibits good control effect against Fusarium pathogens, is safe for crops, delays the development of drug resistance in pathogens, and reduces the dosage of the agent while reducing the pesticide residue in agricultural products.

[0272] Although the present invention has been described in detail above with general description and specific embodiments, some modifications or improvements can be made to it based on the present invention, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention are within the scope of protection claimed by the present invention.

Claims

1. A bactericidal composition, characterized in that, The bactericidal composition comprises active ingredient A and active ingredient B, wherein active ingredient A is a compound of formula I: (Formula I), wherein the active ingredient B is selected from any one of isoflucypram, fluopyram, and fluopyram. The active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:32 to 20:

1. The active ingredient B is fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:35~20:1; The active ingredient B is fluoroamide, and the mass ratio of active ingredient A to active ingredient B is 1:20~30:

1.

2. The bactericidal composition according to claim 1, characterized in that, The active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:25 to 15:1; The active ingredient B is fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:22~15:1; The active ingredient B is fluoroamide, and the mass ratio of active ingredient A to active ingredient B is 1:15 to 20:

1.

3. The bactericidal composition according to claim 2, characterized in that, The active ingredient B is isoflucypram, and the mass ratio of active ingredient A to active ingredient B is 1:18 to 10:

1. The active ingredient B is fluopyram, and the mass ratio of active ingredient A to active ingredient B is 1:16 to 10:1; The active ingredient B is fluoroamide, and the mass ratio of active ingredient A to active ingredient B is 1:5 to 18:

1.

4. The bactericidal composition according to claim 1, characterized in that, The total weight of the bactericidal composition is calculated as 100 wt%, and the total weight of active ingredient A and active ingredient B accounts for 1% to 80% of the total weight of the bactericidal composition.

5. The bactericidal composition according to claim 1, characterized in that, The bactericidal composition includes, in addition to the active ingredient, other auxiliary ingredients selected from one or more of the following: wetting agents, dispersants, emulsifiers, thickeners, disintegrants, antifreeze agents, defoamers, solvents, preservatives, stabilizers, warning colors, film-forming agents, synergists, and carriers.

6. The bactericidal composition according to claim 1, characterized in that, The bactericidal composition can be prepared into any agriculturally acceptable formulation, which may be a solid or liquid formulation. The solid formulation is a water-dispersible granule and / or a wettable powder, and the liquid formulation is a suspension.

7. The use of the bactericidal composition according to any one of claims 1-6 in the prevention or control of crop diseases, characterized in that, The diseases mentioned are wheat sheath blight, rice sheath blight, and cucumber downy mildew.

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