A bactericidal composition and its application

Abstract

The present invention belongs to the field of pesticide sterilization technology and discloses a sterilization composition and its use. The sterilization composition comprises active ingredient A and active ingredient B, wherein active ingredient A is cyclobutrifluram and active ingredient B is any one of ipconazole, metconazole, or trichlorfonazole. The mass ratio of active ingredient A to active ingredient B is 1:50 to 55:1. The sterilization composition of the present invention has a strong control effect on diseases caused by Fusarium fungi, with significant synergistic effects, reduces the frequency and dosage of pesticide use, reduces production costs, and reduces environmental pollution.

Description

Technical Field

[0001] The present invention belongs to the field of pesticide compounding, and in particular relates to a bactericidal composition and application thereof. Background Art

[0002] Triazole fungicides are a class of highly effective fungicides developed in the 1970s. They exert their efficacy by inhibiting the biosynthesis of ergosterol in bacteria. They have good systemic properties, are highly effective, low in toxicity, have a long lasting effect, and have both preventive and therapeutic effects. They can be used to prevent and control various diseases caused by ascomycetes, basidiomycetes and deuteromycetes.

[0003] Cyclobutrifluram is a new nicotinamide fungicide and nematocide developed by Syngenta. IUPAC name: N-[2-(2,4-dichlorophenyl)cyclobutyl]-2-(trifluoromethyl)nicotinamide, containing 80%-100% (1S,2S)-enantiomer and 0-20% (1R,2R)-enantiomer; CAS registration number: 1460292-16-3; molecular formula: C 17 H 13 CI2F3N2O. Cyclobutrifluram has a broad spectrum of control, is highly effective, requires minimal dosage, and is easy to use. It effectively controls various nematodes and major fungal diseases. It can be used as a soil or seed treatment, providing long-term control of nematodes and diseases on major crops and in a variety of environments, and is particularly effective against Fusarium.

[0004] Fusarium fungi are important pathogens that cause a variety of soil-borne diseases of crops. In agricultural production, they can cause a variety of plant diseases, such as wheat ergot, wheat stem rot, rice seedling blight, and other diseases. When severe diseases occur, they cause significant economic losses to agricultural production.

[0005] Wheat stem rot, caused by various Fusarium spp. species, is a worldwide soil-borne disease that manifests as brown rot at the base of the stem, disease, and browning of the sheaths and base of the wheat buds. During the grain-filling period, the tillering of the stem base and the death of the upper portion occur. The stems, leaves, and ears, due to lack of water, die, resulting in withered white ears. These ears easily break at the base when pulled out in the field. Mildly infected plants grow thin and weak, eventually dying, significantly impacting yield. Wheat stem rot is insidious and difficult to detect during the seedling stage. By the time symptoms appear aboveground, it's too late to prevent and control it. Failure to promptly prevent and control it can lead to significant yield losses or even total crop failure, often known as "wheat cancer." Research has found that there are no highly resistant wheat stem rot varieties currently in production, and crop rotation is difficult to implement. Consequently, there is a lack of effective control methods for stem rot.

[0006] Rice bakanae disease, also known as leggy growth disease and white stalk disease, is a seed-borne disease of rice caused by the fungus Fusarium spp. It can typically reduce rice yields by 5% to 20%, with severe cases exceeding 50%. When infected, rice plants begin to grow leggy at the 2- to 4-leaf stage, becoming taller than healthy plants. Their leaves and sheaths are narrow and elongated, and their root systems develop abnormally, resulting in weak and thin growth. Affected plants typically fail to produce or only produce a complete ear. Even when they do produce, the ears are small and unsturdy. In recent years, rice bakanae disease has been on the rise in major rice-producing areas in my country, with severe outbreaks in some regions, posing a threat to stable and high rice yields.

[0007] Determining the biological activity of compounds against target organisms and combining different compounds is an effective method for developing pesticides, improving control effectiveness, and delaying drug resistance. The applicant conducted in-depth research on cyclobutrifluram with ipconazole, metconazole, or triticonil, and on combinations of cyclobutrifluram with any of these. The applicant discovered that mixing cyclobutrifluram with any of these, metconazole, or triticonil, within a certain mixing ratio range, exhibited a significant synergistic effect against Fusarium fungi, effectively improving the control of soil-borne diseases caused by Fusarium fungi. Further research led to the completion of the present invention. Currently, there are no reports on the use of cyclobutrifluram mixed with any of these, metconazole, or triticonil, for controlling diseases caused by Fusarium fungi. Summary of the Invention

[0008] Based on the above problems, the present invention provides a fungicide composition, which can effectively control diseases caused by Fusarium fungi, has obvious synergistic effects, effectively reduces the dosage of pesticides, and is environmentally friendly.

[0009] To achieve the above object, the present invention adopts the following technical solution: a fungicide composition, comprising an active ingredient A and an active ingredient B, wherein the active ingredient A is Cyclobutrifluram, and the active ingredient B is any one of ipconazole, metconazole, or trichlorfonazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:50 to 55:1;

[0010] Furthermore, the active ingredient B is ipconazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:50 to 48:1;

[0011] Furthermore, the active ingredient B is ipconazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:30 to 48:1;

[0012] Furthermore, the active ingredient B is metconazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:40 to 55:1;

[0013] Furthermore, the active ingredient B is metconazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:30 to 48:1;

[0014] Furthermore, the active ingredient B is trichlorfonazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:32 to 48:1;

[0015] Furthermore, the active ingredient B is trichlorfonazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:25 to 30:1;

[0016] Furthermore, the total weight of the bactericidal combination is calculated as 100 wt%, and the total weight of the active ingredient A and the active ingredient B accounts for 0.1% to 80% of the total weight of the bactericidal composition;

[0017] Furthermore, the fungicidal composition contains, in addition to the active ingredient, agriculturally acceptable auxiliary ingredients, wherein the auxiliary ingredients are selected from one or more of a wetting agent, a dispersant, an emulsifier, a thickener, a disintegrant, an antifreeze agent, a defoaming agent, a solvent, a preservative, a stabilizer, a synergist, a binder, a filler or a carrier;

[0018] Furthermore, the wetting agent is selected from one or more of alkylbenzene sulfonate, alkylnaphthalene sulfonate, lignin sulfonate, sodium lauryl sulfate, sodium dioctyl sulfosuccinate, α-olefin sulfonate, alkylphenol polyoxyethylene ether, castor oil polyoxyethylene ether, alkylphenol ethoxylate, fatty alcohol ethoxylate, fatty alcohol polyoxyethylene ether sodium sulfate, silkworm feces, soapberry powder, soapberry powder, SOPA, detergent, emulsifier 2000 series and wetting penetrant F; and / or

[0019] The dispersant is selected from one or more of lignin sulfonate, alkylnaphthalene sulfonate formaldehyde condensate, naphthalene sulfonate, tristyrylphenol ethoxylate phosphate, fatty alcohol ethoxylate, alkylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether methyl ether condensate sulfate, fatty amine polyoxyethylene ether, glycerol fatty acid ester polyoxyethylene ether, polycarboxylates, polyacrylic acids, phosphates, EO-PO block copolymers and EO-PO graft copolymers; and / or

[0020] The emulsifier is selected from one or more of calcium dodecylbenzenesulfonate, alkylphenol formaldehyde resin polyoxyethylene ether, phenylethylphenol polyoxyethylene polyoxypropylene ether, fatty alcohol ethylene oxide-propylene oxide copolymer, styrylphenol polyoxyethylene ether, castor oil polyoxyethylene ether and alkylphenol ether phosphate; and / or

[0021] The thickener is selected from one or more of xanthan gum, organobentonite, gum arabic, sodium alginate, magnesium aluminum silicate, carboxymethyl cellulose and white carbon black; and / or

[0022] Disintegrants: The disintegrants are selected from one or more of sodium sulfate, ammonium sulfate, aluminum chloride, sodium chloride, ammonium chloride, bentonite, glucose, sucrose, starch, cellulose, urea, sodium carbonate, sodium bicarbonate, citric acid and tartaric acid; and / or

[0023] The antifreeze agent is selected from one or more of alcohols, alcohol ethers, chlorinated hydrocarbons and inorganic salts; and / or

[0024] Defoaming agent selected from C 10 -C 20 Saturated fatty acid compounds, silicone oil, silicone compounds, C8-C 10 One or more of fatty alcohols; and / or

[0025] The solvent is selected from one or more of benzene, toluene, xylene, durene, methanol, ethanol, isopropanol, n-butanol, dimethyl sulfoxide, dimethylformamide, cyclohexanone, alkylene carbonate, diesel, solvent oil, vegetable oil, vegetable oil derivatives and water; and / or

[0026] The preservative is selected from one or more of propionic acid, sodium propionic acid, sorbic acid, sodium sorbic acid, potassium sorbic acid, benzoic acid, sodium benzoic acid, sodium p-hydroxybenzoic acid, methyl p-hydroxybenzoate, kasone and 1,2-benzisothiazolin-3-one; and / or

[0027] The stabilizer is selected from one or more of disodium hydrogen phosphate, oxalic acid, succinic acid, adipic acid, borax, 2,6-di-tert-butyl-p-cresol, triethanolamine oleate, epoxidized vegetable oil, kaolin, bentonite, attapulgite, white carbon black, talc, montmorillonite and starch; and / or

[0028] Synergists are selected from synergist, piperonyl butoxide; and / or

[0029] The carrier is selected from one or more of ammonium salts, ground natural minerals, ground artificial minerals, silicates, resins, waxes, solid fertilizers, water, organic solvents, mineral oils, vegetable oils and vegetable oil derivatives.

[0030] The present invention optimizes the content of active ingredients and adjuvants in the pesticide composition to achieve a better balance between toxicity and residue, thereby enhancing efficacy, reducing dosage and lowering costs.

[0031] Furthermore, the fungicide composition can be prepared into a formulation acceptable for pesticides, and the formulation is a seed treatment suspension, a seed treatment dry powder, a microemulsion, an aqueous emulsion, a suspension, a dispersible oil suspension, a soluble concentrate, an emulsifiable concentrate, a suspoemulsion, a microcapsule suspension, a water-dispersible granule, a wettable powder, or a granule.

[0032] Furthermore, the formulation is in the form of a seed treatment suspension, microemulsion, emulsifiable concentrate, suspension, aqueous emulsion, or water-dispersible granule.

[0033] The present invention also discloses the use of the above-mentioned fungicidal composition in preventing and controlling diseases caused by Fusarium fungi.

[0034] Furthermore, the Fusarium fungus is Fusarium oxysporum, Fusarium solani, Fusarium moniliforme, Fusarium graminearum, Fusarium pseudograminearum, Fusarium equisetifolia, Fusarium genus, Fusarium avenae, and Fusarium lamellifolia;

[0035] Furthermore, the Fusarium fungus is Fusarium moniliforme, Fusarium graminearum, Fusarium pseudograminearum, Fusarium equisetum, and Fusarium genus flavum.

[0036] The beneficial effects of the present invention are as follows:

[0037] 1) The pesticide composition of the present invention has a significant effect on delaying the development of resistance in pathogens by rationally compounding active ingredients with different mechanisms of action;

[0038] 2) The pesticide composition of the present invention has an excellent control effect on Fusarium fungi, a significant synergistic effect, and effectively reduces the damage of Fusarium fungi to plants. DETAILED DESCRIPTION

[0039] In order to make the objects, advantages and technical solutions of the present invention more clearly understood, the present invention uses the following preparation examples and specific embodiments to explain the technical solutions of the present invention. However, the scope of protection of the present invention should not be limited to the specific implementation methods described herein.

[0040] Preparation example:

[0041] Preparation Example 1: 16% Cyclobutrifluram·Ipconazole seed treatment suspension (1:3)

[0042] Formula composition: Cyclobutrifluram 4%, ipconazole 12%, isotridecyl alcohol polyoxyethylene ether 1%, styrylphenol polyoxyethylene ether phosphate 2%, sodium lignin sulfonate 2%, polyacrylic acid emulsion 1%, xanthan gum 0.2%, rose red pigment 5%, ethylene glycol 5%, magnesium aluminum silicate 1%, silicone defoamer 0.5%, sodium benzoate 1%, and deionized water to make up the balance;

[0043] Preparation method: According to the ratio, the active ingredient, adjuvant and water are mixed and stirred evenly through high shear, and then sanded for 2.5 hours in a sander to make the average particle size reach 1 to 5 microns to obtain the seed treatment suspension.

[0044] Preparation Example 2: 9% Cyclobutrifluram·Ipconazole Microemulsion (1:5)

[0045] Formula composition: Cyclobutrifluram 1.5%, ipconazole 7.5%, xylene 13%, cyclohexanone 20%, alkylphenol polyoxyethylene ether 12%, EO-PO block copolymer 3%, styrenated phenol polyoxyethylene ether sodium sulfate 2%, ethylene glycol 5%, silicone defoamer 0.05%, and deionized water to make up the balance;

[0046] Preparation method: Active ingredients, solvents, emulsifiers, etc. are mixed to prepare an oil phase, antifreeze solution and water are mixed to prepare an aqueous phase, the oil phase is added to the aqueous phase under stirring and stirred evenly, shearing is continued for 10 minutes, and then a silicone defoaming agent is added and stirred evenly to obtain oil phase particles with small droplets of 0.01 to 0.1 microns, thereby preparing the microemulsion described in the present invention.

[0047] Preparation Example 3: 12% Cyclobutrifluram·Ipconazole EC (7:1)

[0048] Formula composition: Cyclobutrifluram 10.5%, ipconazole 1.5%, propylene carbonate 12%, calcium dodecylbenzenesulfonate 4%, Guerbet alcohol polyoxyethylene ether 10%, N,N-dimethylformamide 15%, cyclohexanone 15%, and solvent oil to make up the balance;

[0049] Preparation method: add the measured active ingredients, solvent and cosolvent into a mixing kettle and stir to dissolve them, then add emulsifier, make up the balance with the remaining solvent, stir evenly in a stirring kettle, and filter to obtain the desired emulsifiable concentrate of the present invention.

[0050] Preparation Example 4: 12% Cyclobutrifluram·Metconazole Suspension Concentrate (5:1)

[0051] Formula composition: Cyclobutrifluram 10%, metconazole 2%, sodium dioctyl sulfosuccinate 3%, EO-PO block copolymer 2%, sodium sulfate of alkylphenol polyoxyethylene ether methyl ether condensate 3%, sodium polycarboxylate 1%, silicone defoamer 0.5%, xanthan gum 0.2%, magnesium aluminum silicate 1%, ethylene glycol 5%, sodium benzoate 0.5%, and deionized water to make up the balance;

[0052] Preparation method: According to the formula ratio, the active ingredients, surfactants and other functional additives are placed in a reactor in sequence, water is added and mixed evenly, and the suspension product is obtained by high-speed shearing, wet sand grinding, and finally homogenization filtration.

[0053] Preparation Example 5: 21% Cyclobutrifluram·Metconazole Water Dispersible Granules (1:6)

[0054] Formula composition: Cyclobutriflura 3%, metconazole 18%, sodium lauryl sulfate 2%, naphthalenesulfonate formaldehyde condensate 8%, sodium polycarboxylate 2%, white sugar 5%, kaolin makes up the balance;

[0055] Preparation method: According to the formula ratio of the embodiment, the active ingredient is added to the carrier, and a surfactant and other functional additives are added thereto, mixed, and after air flow grinding, 10-25% water is added, and then kneading, granulation, drying, and sieving are carried out to obtain a water-dispersible granule product; or the pulverized powder is sprayed with water in a boiling granulator, granulated, dried, and then sieved to obtain the product.

[0056] Preparation Example 6: 12% Cyclobutrifluram·Metconazole Emulsion in Water (1:1)

[0057] Formula composition: Cyclobutrifluram 6%, metconazole 6%, glycerol fatty acid ester polyoxyethylene ether 3%, polyoxyethylene sorbitan monooleate 3%, cyclohexanone 20%, xanthan gum 0.2%, ethylene glycol 5%, urea 2%, sodium sorbate 0.2%, silicone defoamer 0.5%, deionized water to make up the balance;

[0058] Preparation method: According to the formula ratio of the example, the active ingredient is dissolved in the solvent and an emulsifier is added to dissolve it into a uniform oil phase. Deionized water, antifreeze, etc. are mixed together to form a uniform water phase; under high-speed shear, the oil phase is added to the water phase, and finally a thickener and defoaming agent are added to form a well-dispersed water emulsion product.

[0059] Preparation Example 7: 18% Cyclobutrifluram·Triticonazole seed treatment suspension (1:8)

[0060] Formula composition: Cyclobutrifluram 2%, trichlorfon 16%, glycerol fatty acid ester polyoxyethylene ether 2%, fatty alcohol ethylene oxide-propylene oxide copolymer 2%, sodium lignin sulfonate 2%, magnesium aluminum silicate 1%, xanthan gum 0.3%, polyacrylic acid emulsion 1%, rose red pigment 5%, propylene glycol 5%, silicone defoamer 0.5%, kason 0.1%, and deionized water to make up the balance.

[0061] Preparation method: Same as Preparation Example 1.

[0062] Preparation Example 8: 21% Cyclobutrifluram·Triticonazole seed treatment suspension (2:1)

[0063] Formula composition: Cyclobutrifluram 14%, trichlorfonazole 7%, sodium octylphenol polyoxyethylene ether sulfonate 1%, sodium polycarboxylate 1%, alkyl aromatic polyoxyethylene polyoxypropylene ether 2%, fatty alcohol polyoxyethylene ether sodium sulfate 3%, magnesium aluminum silicate 1%, xanthan gum 0.2%, polyacrylic acid emulsion 1%, rose red pigment 6%, propylene glycol 5%, silicone defoamer 0.5%, sodium parahydroxybenzoate 0.2%, and deionized water to make up the balance.

[0064] Preparation method: Same as Preparation Example 1.

[0065] Preparation Example 9: 22% Cyclobutrifluram·Triticonazole seed treatment suspension (10:1)

[0066] Formula composition: Cyclobutrifluram 20%, trichlorfonazole 2%, alkylphenol polyoxyethylene ether methyl ether condensate sodium sulfate 1%, alkyl naphthalene sulfonate sodium 1%, alkylphenol polyoxyethylene ether 3%, alkyl polyoxyethylene ether sodium sulfonate 3%, magnesium aluminum silicate 1%, xanthan gum 0.2%, polyacrylic acid emulsion 1%, rose red pigment 6%, glycerol 5%, silicone defoamer 0.5%, benzisothiazolinone potassium 0.1%, and deionized water to make up the balance;

[0067] Preparation method: Same as Preparation Example 1.

[0068] Example 1: Indoor activity test of cyclobutrifluram in combination with any one of ipconazole, metconazole or trichlorfonazole against wheat stem rot Test basis: The test was conducted in accordance with NY / T 1156.2-2006 Agricultural Industry Standard of the People's Republic of China "Guidelines for Indoor Bioassay Tests of Pesticides"

[0069] kill

[0070] Bacterial agents - Part 2: Petri dish method for inhibition of mycelial growth of pathogenic bacteria.

[0071] Test target: Fusarium pseudograminearum.

[0072] Instruments and equipment: moist heat sterilizer, clean workbench, constant temperature light incubator, electric drum windproof drying oven, 1 / 10,000 electronic balance, pipette, alcohol lamp, small beaker, volumetric flask, Erlenmeyer flask, culture dish (Φ9cm), hole punch, inoculator (Φ0.6cm), ruler, etc.

[0073] Test agents: 97% ipconazole technical, 95% metconazole technical, 95% trichlorfonazole technical, 90% cyclobutrifluram technical, all provided by the Group's R&D Center.

[0074] Preparation of pharmaceutical preparation: Dissolve the above raw drugs in acetone respectively, and then dilute them with 0.1% Tween 80 aqueous solution to prepare single-dose stock solutions. Different ratios are designed according to the mixing purpose and the activity of the pharmaceutical preparation. Each single dose and each group of mixed preparations are prepared into the required series of mass concentrations.

[0075] Test repetition: 4 culture dishes were used for each concentration of the test drug, 1 culture dish was used for each repetition, and a total of 4 repetitions were performed. A 0.1% Tween 80 aqueous solution without drug was used as a blank control.

[0076] Drug Treatment: Under aseptic conditions, add pre-thawed, sterilized PDA medium to a sterile Erlenmeyer flask according to the test treatment. Pipette 10 mL of each prepared drug solution, starting from low to high concentration, into each Erlenmeyer flask. Shake thoroughly, then pour equal amounts into culture dishes to prepare drug-containing plates of the corresponding concentrations. A blank control of 0.1% Tween 80 in water without the addition of drug was used. Four replicates were performed for each treatment.

[0077] Inoculation: Cut the pre-cultured Fusarium graminearum from the edge of the colony with a sterilized puncher under sterile conditions, inoculate the cake into the center of the drug-containing plate with an inoculator, cover the plate with a lid, and place it in a constant temperature and light incubator at 26°C for dark culture.

[0078] Data Analysis: Investigate the growth of pathogenic mycelia based on the growth of mycelia in the blank control culture dish. Measure the diameter of the colonies in centimeters (cm) using a ruler. Measure the diameter of each colony once using the cross-hatch method, average the values, and record the raw data for all replicates for each treatment.

[0079] Data statistics and analysis: Based on the survey results, calculate the mycelial growth inhibition rate of each treatment concentration on the target bacteria, the unit is percentage (%), and the calculation results are rounded to two decimal places.

[0080] D=D1-D2

[0081] Where:

[0082] D——colony growth diameter;

[0083] D1 - colony diameter;

[0084] D2——diameter of mushroom cake.

[0085]

[0086] Where:

[0087] I——hyphae growth inhibition rate;

[0088] D0——blank control colony growth diameter;

[0089] D T ——The growth diameter of the colony after treatment with chemicals.

[0090] Use DPS statistical analysis system to analyze and calculate the toxicity regression line and EC 50 The activity of the test agent on the biological test material is evaluated by the value.

[0091] Sun Yunpei method: The synergistic effect of mixed drugs is evaluated based on the co-toxicity coefficient (CTC). A co-toxicity coefficient of CTC ≥ 120 indicates a synergistic effect; CTC ≤ 80 indicates an antagonistic effect; and 80 < CTC < 120 indicates an additive effect.

[0092] Calculation of the co-toxicity coefficient (CTC value) of the mixture:

[0093]

[0094] Where:

[0095] ATI - measured toxicity index of mixture;

[0096] S——EC of standard agent 50 , the unit is milligrams per liter (mg / L);

[0097] M——EC of the mixture 50 , the unit is milligrams per liter (mg / L).

[0098] TTI=TI A ×P A +TI B ×P B

[0099] Where:

[0100] TTI – Theoretical Toxicity Index of Mixtures;

[0101] TI A ——Agent toxicity index;

[0102] P A ——The percentage of agent A in the mixture, in percentage (%);

[0103] TI B ——Toxicity index of agent B;

[0104] P B ——The percentage of agent B in the mixture, in percentage (%).

[0105]

[0106] Where:

[0107] CTC – Co-toxicity coefficient;

[0108] ATI - measured toxicity index of mixture;

[0109] TTI - Theoretical Toxicity Index of Mixture.

[0110] The test results are shown in the table below:

[0111] Table 1 Indoor bioactivity test of cyclobutrifluram and ipconazole against Fusarium graminearum

[0112]

[0113] Table 2 Indoor bioactivity test of Cyclobutrifluram and metconazole against Fusarium graminearum

[0114]

[0115] Table 3 Indoor bioactivity test of Cyclobutrifluram and trichlorfonazole against Fusarium graminearum

[0116]

[0117]

[0118] As shown in Tables 1-3 above, a reasonable combination of cyclobutrifluram with any of the triazole fungicides ipconazole, metconazole, or triticonazole exhibits excellent control effectiveness against F. graminearum. A cyclobutrifluram to ipconazole mass ratio of 1:50 to 48:1 exhibits a synergistic effect against F. graminearum. A cyclobutrifluram to metconazole mass ratio of 1:40 to 55:1 exhibits a synergistic effect. A cyclobutrifluram to triticonazole mass ratio of 1:32 to 48:1 exhibits a synergistic effect against F. graminearum.

[0119] Example 2: Indoor activity test of cyclobutrifluram in combination with any one of ipconazole, metconazole or trichlorfonazole against rice bakanae disease

[0120] Test basis: The test refers to NY / T 1156.2-2006 Agricultural Industry Standard of the People's Republic of China "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 2: Plate Method for Inhibition of Pathogen Mycelial Growth".

[0121] Test strain: Fusarium moniliforme, provided by Shenyang Research Institute of Chemical Industry.

[0122] Instruments and equipment: high-pressure steam sterilizer, clean bench, incubator, electric blast drying oven, 1 / 10,000 electronic balance, pipette, alcohol lamp, beaker (50mL), volumetric flask, Erlenmeyer flask (100mL), culture dish (Φ9cm), hole punch (Φ0.6cm), inoculator, ruler, etc.

[0123] The target culture conditions were as follows: Fusarium moniliforme stored in a refrigerator at 4°C was transferred to potato dextrose agar medium, and cultured in a dark incubator at 25°C for 5 days to activate it and set aside.

[0124] Test agents: 97% ipconazole technical, 95% metconazole technical, 95% trichlorfonazole technical, 90% cyclobutrifluram technical, all provided by the Group's R&D Center.

[0125] Preparation of pharmaceutical preparation: Dissolve the above raw drugs in acetone to prepare high-concentration stock solutions, and then dilute them with 0.1% Tween 80 aqueous solution to prepare single-dose stock solutions. Different ratios are designed according to the mixing purpose and the activity of the pharmaceutical preparation. Each single dose and each group of mixed preparations are prepared into the required series of mass concentrations.

[0126] Test repetition: 4 culture dishes were used for each concentration of the test drug, 1 culture dish was used for each repetition, and a total of 4 repetitions were performed. A 0.1% Tween 80 aqueous solution without drug was used as a blank control.

[0127] Drug treatment: Under aseptic operating conditions, use a pipette to add 5 mL of drug solution of different concentrations to a pre-calibrated sterilized conical flask, then add the culture medium that has been melted and cooled to an appropriate temperature to the conical flask. After shaking thoroughly, pour equal amounts into 4 culture dishes to prepare drug-containing PDA plates of corresponding concentrations.

[0128] Inoculation: Cut the pre-cultured Fusarium moniliforme from the edge of the colony with a sterile punch under sterile conditions, inoculate the cake into the center of the drug-containing plate with an inoculator, cover it with a cap, and place it in a constant temperature incubator at 25°C for dark culture.

[0129] Data analysis: Conduct the test when the colonies in the control treatment grow to 2 / 3 to 4 / 5 of the diameter of the culture dish. Measure the colony diameter (cm) with a ruler. Measure the diameter of each colony once using the cross-hatch method and take the average value.

[0130] Data statistics and analysis: Based on the survey results, calculate the mycelial growth inhibition rate of each treatment concentration on the target bacteria, the unit is percentage (%), and the calculation results are rounded to two decimal places.

[0131] D=D1-D2

[0132] Where:

[0133] D——colony growth diameter;

[0134] D1 - colony diameter;

[0135] D2——diameter of mushroom cake.

[0136]

[0137] Where:

[0138] I——hyphae growth inhibition rate;

[0139] D0——blank control colony growth diameter;

[0140] D T ——The growth diameter of the colony after treatment with chemicals.

[0141] Use DPS statistical analysis system to analyze and calculate the toxicity regression line and EC 50 The activity of the test agent on the biological test material is evaluated by the value.

[0142] Sun Yunpei method: The synergistic effect of mixed drugs is evaluated based on the co-toxicity coefficient (CTC). A co-toxicity coefficient of CTC ≥ 120 indicates a synergistic effect; CTC ≤ 80 indicates an antagonistic effect; and 80 < CTC < 120 indicates an additive effect.

[0143] Calculation of the co-toxicity coefficient (CTC value) of the mixture:

[0144]

[0145] Where:

[0146] ATI - measured toxicity index of mixture;

[0147] S——EC of standard agent 50 , the unit is milligrams per liter (mg / L);

[0148] M——EC of the mixture 50 , the unit is milligrams per liter (mg / L).

[0149] TTI=TI A ×P A +TI B ×P B

[0150] Where:

[0151] TTI – Theoretical Toxicity Index of Mixtures;

[0152] TI A ——Agent toxicity index;

[0153] PA ——The percentage of agent A in the mixture, in percentage (%);

[0154] TI B ——Toxicity index of agent B;

[0155] P B ——The percentage of agent B in the mixture, in percentage (%).

[0156]

[0157] Where:

[0158] CTC – Co-toxicity coefficient;

[0159] ATI - measured toxicity index of mixture;

[0160] TTI - Theoretical Toxicity Index of Mixture.

[0161] The test results are shown in the table below:

[0162] Table 4 Indoor bioactivity test of Cyclobutrifluram and ipconazole against Fusarium moniliforme

[0163]

[0164] Table 5 Indoor bioactivity test of Cyclobutrifluram and metconazole against Fusarium moniliforme

[0165]

[0166]

[0167] Table 6 Indoor bioactivity test of Cyclobutrifluram and trichlorfonazole against Fusarium moniliforme

[0168]

[0169] As shown in Tables 4-6 above, a reasonable combination of cyclobutrifluram with any of the triazole fungicides ipconazole, metconazole, or triticonazole exhibits excellent control effectiveness against Fusarium moniliforme. A cyclobutrifluram to ipconazole mass ratio of 1:30 to 48:1 exhibits a synergistic effect against Fusarium moniliforme. A cyclobutrifluram to metconazole mass ratio of 1:45 to 40:1 exhibits a synergistic effect. A cyclobutrifluram to triticonazole mass ratio of 1:25 to 30:1 exhibits a synergistic effect against Fusarium moniliforme.

[0170] Example 3: Field test on controlling rice bakanae disease

[0171] Test basis: The test refers to GB / T 17980.104-2004 "Guidelines for Field Efficacy Tests of Pesticides (II) Part 104: Bactericidal and Control of Rice Bakanae Disease".

[0172] Test target: Rice seedling pathogen.

[0173] Experimental crops: rice.

[0174] Experimental design: The plots of experimental agent, control agent and blank control were arranged in random blocks. The experiment set up 7 agent soaking treatments and used water as blank control. The area of each seedbed treatment was 10m 2 , the field planting area is 30m 2 , each treatment was repeated 4 times.

[0175] The experiment involved soaking seeds on May 31, 2022, germinating on June 2, and sowing on June 5. Each treatment solution was prepared according to the experimental concentration and thoroughly stirred. Dry seeds were then added and soaked for 48 hours to accelerate germination. During the soaking period, the seeds were kept out of the water. Three days later, the seeds were sown in seedling trays. Seedling bed temperature and humidity were strictly controlled according to rice technical regulations. After emergence, the seedling trays were moved to the nursery field, and the seedlings were transplanted to the field at 30 days old.

[0176] Survey items: When all rice seedlings have emerged on the seedbed, the emergence rate of each treatment plot will be investigated. The disease index survey will be conducted once before transplanting and once during the heading stage of rice in the field.

[0177] Before transplanting the seedlings, samples were taken at 5 points in each plot in the seedling field, 200 seedlings were surveyed at each point, the disease rate was investigated, and the control effect was calculated;

[0178] During the field investigation at the booting stage, 5 random sampling points were taken in each plot, 20 clusters were investigated at each point, the number of diseased plants was recorded, and the diseased plant rate and control effect of each treatment were calculated.

[0179] Calculation method of drug efficacy:

[0180]

[0181]

[0182]

[0183] The test results are shown below:

[0184] Table 7 Field test results for controlling rice bakanae disease

[0185]

[0186]

[0187] As shown in Table 7, the pesticide compositions of the present invention all exhibited significant control efficacy against rice bakanae disease. Compared to the blank control, the seedling emergence rates were essentially equivalent, indicating that soaking rice seeds with each formulation did not cause phytotoxicity. Furthermore, compared to single-agent fungicides, each combined formulation exhibited a reduced diseased plant rate, demonstrating superior control efficacy.

[0188] Example 4: Field test on prevention and control of wheat stem rot

[0189] Test target: wheat stem rot pathogen.

[0190] Experimental crop: wheat.

[0191] Application method: There are two methods of application: root spraying and seed mixing.

[0192] Experimental design: The corresponding chemical treatments were set according to the experimental method; 5 treatments were set for the root spray test and 8 treatments were set for the chemical seed dressing test. The above experiments were repeated 4 times. Each experimental plot was arranged in a randomized block group with an area of 20m 2 .

[0193] Test method: The root spray test was carried out 3 and a half months after wheat planting. The root spray was carried out at the early stage of wheat stem base rot. The spray was applied again after an interval of 7 days, for a total of 2 applications.

[0194] Seed dressing with pesticides: Wheat seeds were treated with pesticides according to the experimental design before planting.

[0195] Survey methods and time:

[0196] The experiment was conducted during the wheat milk stage. Five random sampling points were selected in each plot, with 100 plants selected at each point. The number of diseased plants at each level was investigated. The grading standards are as follows:

[0197] The condition is graded according to the following criteria:

[0198] Level 0: No browning symptoms on the whole plant;

[0199] Level 1: The roots are browning;

[0200] Level 3: The first node of the aboveground part turns brown and rots;

[0201] Level 5: The second node of the aboveground part turns brown and rots;

[0202] Level 7: The lesions extend beyond the second stem node, but there is no white spike;

[0203] Level 9: The lesions extend beyond the second stem node and there are white spikes;

[0204] Calculation formula for disease index and prevention and treatment effect:

[0205]

[0206]

[0207] The test results are shown below:

[0208] Table 8 Wheat stem rot pesticide seed dressing test results

[0209]

[0210] Table 9 Wheat stem rot root spray test results

[0211]

[0212] Field efficacy showed that the mixture of Cyclobutrifluram with ipconazole, metconazole and trichlorfonazole showed significantly better antibacterial activity against wheat stem rot than any single agent.

[0213] In summary, through indoor toxicity testing and field efficacy tests, it can be seen that the pesticide composition of the present invention has a good control effect on Fusarium, is safe for target crops, has significant control effects, is superior to a single dose in delaying the development of drug resistance and prolonging the effectiveness, can effectively reduce costs and reduce drug residues.

[0214] Although the present application describes specific embodiments in detail with the aid of examples, the disclosure of the present application may adopt various modifications and alternative forms. However, it should be understood that the disclosure of the present application is not limited to the specific forms disclosed. On the contrary, the disclosure of the present application covers all modifications, equivalents and alternative forms within the scope of the disclosure of the present application, and the scope of the present application is limited by the appended claims and their legal equivalents.

Claims

1. A bactericidal composition, characterized in that The fungicidal composition comprises active ingredient A and active ingredient B, wherein the active ingredient A is cyclobutrifluram, the active ingredient B is ipconazole, and the mass ratio of the active ingredient A to the active ingredient B is 1:50 to 48:

1.

2. The bactericidal composition according to claim 1, characterized in that The mass ratio of the active ingredient A to the active ingredient B is 1:30-48:

1.

3. The bactericidal composition according to claim 1, characterized in that The total weight of the bactericidal combination is calculated as 100 wt %, and the total weight of the active ingredient A and the active ingredient B accounts for 0.1% to 80% of the total weight of the bactericidal composition.

4. The bactericidal composition according to claim 1, characterized in that In addition to the active ingredients, the fungicidal composition also contains agriculturally permitted auxiliary ingredients, and the auxiliary ingredients are selected from one or more of wetting agents, dispersants, emulsifiers, thickeners, disintegrants, antifreeze agents, defoaming agents, solvents, preservatives, stabilizers, synergists, binders, fillers or carriers.

5. The bactericidal composition according to claim 1, characterized in that The fungicide composition can be prepared into a formulation acceptable for pesticides, and the formulation is a seed treatment suspension, a seed treatment dry powder, a microemulsion, an aqueous emulsion, a suspension, a dispersible oil suspension, a soluble solution, an emulsifiable concentrate, a suspoemulsion, a microcapsule suspension, a water-dispersible granule, a wettable powder, or a granule.

6. The bactericidal composition according to claim 5, characterized in that The preparation dosage forms include seed treatment suspension, microemulsion, emulsifiable concentrate, suspension, aqueous emulsion and water dispersible granules.

7. Use of the fungicidal composition according to any one of claims 1 to 6 for preventing and controlling diseases caused by Fusarium fungi, characterized in that: The Fusarium fungi are Fusarium moniliforme and Fusarium graminearum.

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