Use of a fungicidal composition containing benzovindiflucarb and pyraclostrobin for controlling plant guignardia diseases

Abstract

The application belongs to the field of pesticide fungicides, and discloses a use of a fungicidal composition containing benzoic fluoroazoles and pyraclostrobin for preventing and treating plant colletotrichum diseases; active ingredients of the fungicidal composition include active ingredient A benzoic fluoroazoles and active ingredient B pyraclostrobin. The fungicidal composition can be used for preventing and treating diseases caused by colletotrichum on plants, has good prevention and treatment effect, and has the characteristics of wide fungicidal spectrum, low drug dosage, obvious synergistic effect and the like.

Description

Technical Field

[0001] This invention relates to the field of pesticides and fungicides, specifically to the use of a fungicide composition containing benzylfloxacin and pyraclostrobin for the prevention and control of anthracnose in plants. Background Technology

[0002] Benzoflubenzuron is a pyrazole amide fungicide developed by Syngenta. It is a succinate dehydrogenase inhibitor and is widely used to control various diseases, including wheat powdery mildew and Asian soybean rust. It exhibits no cross-resistance with methoxyacrylate and triazole fungicides and can be used in combination with a variety of fungicides. Its chemical structure is as follows:

[0003]

[0004] Pyraclostrobin is a methoxyacrylate fungicide, a mitochondrial respiration inhibitor, and possesses protective, curative, and leaf transdermal effects. In addition to its direct action against various pathogens, this fungicide also promotes crop growth and increases yield.

[0005] Existing technology CN108094433A discloses a fungicide composition, specifically disclosing that benzimidazole and pyraclostrobin have a good synergistic effect against rice sheath blight in the range of 1:30 to 30:1; CN104839165A discloses a fungicide composition of benzimidazole, further disclosing that the mass ratio of benzimidazole to pyraclostrobin is 1:20 to 20:1, and further disclosing the application of the aforementioned fungicide composition of benzimidazole for the control of crops. However, there are no relevant reports on the use of benzimidazole and pyraclostrobin for the control of plant anthracnose. Summary of the Invention

[0006] Based on the above, the purpose of this invention is to provide a fungicide composition containing benzimidazole and pyraclostrobin for the control of plant anthracnose diseases. This fungicide composition has good control effect and features a broad fungicide spectrum, low dosage, and significant synergistic effect.

[0007] To achieve the above objectives, the following technical solution is provided: a fungicide composition containing benzimidazole and pyraclostrobin for the prevention and control of plant anthracnose diseases, wherein the active ingredients of the fungicide composition include active ingredient A benzimidazole and active ingredient B pyraclostrobin, and the mass ratio of benzimidazole to pyraclostrobin is 1:35 to 25:1.

[0008] Furthermore, the mass ratio of the active ingredient benzimidazole to pyraclostrobin is 1:24 to 25:1;

[0009] Furthermore, the mass ratio of the active ingredient benzimidazole to pyraclostrobin is 1:10 to 14:1;

[0010] Furthermore, based on a total weight of 100 wt% of the bactericidal composition, the sum of the contents of benzo[a]flufenicol and pyraclostrobin in the bactericidal composition is 5 to 90 wt%.

[0011] Furthermore, the sum of the contents of benzo[i]flufenican and pyraclostrobin in the bactericidal composition is 5-80 wt%.

[0012] Furthermore, the formulation of the bactericidal composition can be any one of emulsifiable concentrate, water emulsion, microemulsion, suspension concentrate, wettable powder or water-dispersible granules;

[0013] Furthermore, the aforementioned plant anthracnose is anthracnose caused by fungi of the genus *Colletotrichum*.

[0014] Furthermore, the aforementioned anthrax fungi include *Colletotrichum higginsianum*, *Colletotrichum orbiculare*, *Colletotrichum lindemuthianum*, *Colletotrichum capsici*, and *Colletotrichum gloeosporioides (Penz.) Penz. et Sacc*.

[0015] Furthermore, the plant diseases caused by the anthracnose fungus are cucumber anthracnose [Colletotrichum orbiculare] and citrus anthracnose [Colletotrichum gloeosporioides (Penz.) Penz. et Sacc].

[0016] Furthermore, the bactericidal composition is applied in an effective dose to the disease or its growth medium that needs to be controlled.

[0017] Compared with the prior art, the beneficial effects of the technical solution of the present invention are as follows:

[0018] 1) The combination of benzimidazole and pyraclostrobin increases the fungicidal activity and can effectively control anthracnose diseases in plants.

[0019] 2) Safe and environmentally friendly, with broad development prospects, reducing the amount of pesticides used and lowering farmers' pesticide costs;

[0020] 3) It is highly efficient, has a broad bactericidal spectrum, and can delay the development of drug resistance in harmful bacteria and prolong the duration of drug efficacy. Detailed Implementation

[0021] To make the technical solution, objectives and advantages of the present invention clearer, the present invention is described with reference to the following specific embodiments. However, the present invention can be implemented in various forms and should not be limited to the embodiments described herein.

[0022] Indoor toxicity testing

[0023] Example 1

[0024] Indoor combined effect test of benzimidazole and pyraclostrobin on cucumber anthracnose

[0025] Test basis: The test references NY / T 1156.2-2006 "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 2: Plate Method for Inhibition of Mycelial Growth of Pathogenic Fungi" and NY / T 1156.6-2006 "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 6: Determination of Combined Effects of Mixtures".

[0026] Test reagents: 96% benzo[i]flufenicol technical grade and 97% pyraclostrobin technical grade, provided by the Group's R&D Center.

[0027] The tested pathogen was *Colletotrichum lagenarium* (Pass.) Ell. ethalst.

[0028] Reagent preparation: Dissolve the test drug in acetone first, then dilute with 0.1% Tween 80 aqueous solution. Prepare single-agent stock solutions separately, and set 5 series of mass concentrations according to the purpose of mixing and the drug activity. The final content of organic solvent shall not exceed 2%.

[0029] Melt the PDA medium in a microwave oven and cool it to about 50°C. Following the principle of starting from low concentration and gradually increasing the concentration, take 1 mL of the prepared test solution and 9 mL of PDA medium and add them to a 9 cm diameter petri dish. Mix well to prepare a drug-containing plate of the corresponding concentration.

[0030] Inoculation: Under aseptic conditions, the activated pathogenic fungi were punched into a fungal cake using a 6mm diameter punch. After the drug-containing culture medium solidified, the fungal cake was placed in the center of the culture medium. Finally, the culture dish was sealed with sealing film and placed in an incubator at 27℃ for incubation. A blank solution without the drug was set up as a blank control. Each treatment was repeated 4 times.

[0031] Investigation: After 12 days of cultivation, the diameter of the colonies was measured with calipers in millimeters (mm). The diameter of each colony was measured vertically once using the cross-sectional method, and the average value was taken.

[0032] Data statistics and analysis: The growth of pathogenic mycelia was investigated based on the growth of bacteria in the blank control culture dishes. The colony diameter was measured using the cross-crossing method.

[0033] Based on the survey results, the inhibition rate of mycelial growth of the tested target bacteria by each treatment concentration was calculated as a percentage (%), and the calculation results were retained to two decimal places.

[0034] D = D1 - D2

[0035] In the formula:

[0036] D – Colony growth diameter;

[0037] D1—colony diameter;

[0038] D2 – Diameter of the mushroom cake.

[0039]

[0040] I – Mycelial growth inhibition rate;

[0041] D0—Correlation diameter of the blank control group;

[0042] D t — Diameter of colonies grown after chemical treatment.

[0043] Experimental statistics: Data were processed using probability value analysis. The DPS statistical analysis system was used to analyze the data and determine the toxicity regression line, R, and EC. 50 Value (95% confidence interval), EC 90 Values ​​(95% confidence interval) are used to evaluate the activity of the test reagent on the biological sample.

[0044] The co-toxicity coefficient (CTC value) of the mixture is calculated using the following formula:

[0045]

[0046] In the formula:

[0047] ATI – Actual Measured Toxicity Index of Mixtures;

[0048] S – EC of standard bactericides 50 The unit is milligrams per liter (mg / L);

[0049] M – EC of the mixture 50 The unit is milligrams per liter (mg / L).

[0050] TTI = TI A *P A +TI B *P B

[0051] In the formula:

[0052] TTI – Theoretical Toxicity Index of Mixtures;

[0053] TI A —A. Toxicity index of drug A;

[0054] P A —Percentage content of drug A in the mixture, expressed as percentage (%);

[0055] TI B —Toxicity index of drug B;

[0056] P B —Percentage content of agent B in the mixture, expressed as percentage (%).

[0057]

[0058] In the formula:

[0059] CTC – Cotoxicity Coefficient;

[0060] ATI – Actual Measured Toxicity Index of Mixtures;

[0061] TTI – Theoretical Toxicity Index of Mixtures.

[0062] 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.

[0063] The test results are shown in the table below:

[0064] Indoor activity tests showed (see Table 1) that the mixture of benzimidazole and pyraclostrobin was effective against the cucumber anthracnose pathogen EC. 50 The concentrations of pyraclostrobin and benzimidoxam were 0.765 mg / L and 0.1530 mg / L, respectively, indicating that the cucumber anthracnose pathogen was relatively sensitive to pyraclostrobin. The co-toxicity coefficients of benzimidoxam and pyraclostrobin at a ratio of 1:35–25:1 were all greater than 80, showing an additive or synergistic effect in controlling cucumber anthracnose. The co-toxicity coefficients of benzimidoxam and pyraclostrobin at a ratio of 1:24–25:1 were all greater than 120, showing a synergistic effect in controlling cucumber anthracnose. The highest co-toxicity coefficient (173.08) was achieved when benzimidoxam and pyraclostrobin were mixed at a ratio of 3:1. 50 It is 0.221 mg / L.

[0065] Table 1. Results of the indoor combined effect test of benzo[a]fluopyram and pyraclostrobin on cucumber anthracnose.

[0066]

[0067] Example 2

[0068] Indoor combined effect test of benzimidazole and pyraclostrobin on citrus anthracnose

[0069] Test basis: The test references NY / T 1156.2-2006 "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 2: Plate Method for Inhibition of Mycelial Growth of Pathogenic Fungi" and NY / T 1156.6-2006 "Guidelines for Indoor Bioassay of Pesticides - Fungicides Part 6: Determination of Combined Effects of Mixtures".

[0070] Test strain: Colletotrichum gloeosporioides, the causal agent of citrus anthracnose.

[0071] Test reagents: 96% benzo[i]flufenicol technical grade and 97% pyraclostrobin technical grade, provided by the Group's R&D Center.

[0072] Drug preparation: The original drug is first dissolved in acetone to prepare single-dose mother liquors. Different ratios are designed according to the purpose of mixing and drug activity. Each single drug and each group of mixed solutions are prepared into the required series of mass concentration gradients.

[0073] Under aseptic conditions, pre-melted sterile PDA medium was cooled to 50°C. Using a pipette, 1 mL of the drug solution was mixed thoroughly with 9 mL of PDA medium in a 9 cm diameter petri dish to prepare drug-coated agar plates. Sterile water was used as a blank control. Each treatment was repeated four times.

[0074] Inoculation: Activate the preserved pathogens, and under aseptic conditions, use a 6mm diameter sterile punch to cut off the mycelial cake from the edge of the colony. Use an inoculator to inoculate the mycelial cake into the center of the drug-containing agar plate, with the mycelial side facing up. Place one mycelial cake in each petri dish, cover the dish, and incubate in a constant temperature incubator at 25℃.

[0075] Data statistics and analysis: The growth of pathogenic mycelia was investigated based on the growth of bacteria in the blank control culture dishes. The colony diameter was measured using the cross-crossing method.

[0076] Based on the survey results, the inhibition rate of mycelial growth of the tested target bacteria by each treatment concentration was calculated as a percentage (%), and the calculation results were retained to two decimal places.

[0077] D = D1 - D2

[0078] In the formula:

[0079] D – Colony growth diameter;

[0080] D1—colony diameter;

[0081] D2 – Diameter of the mushroom cake.

[0082]

[0083] I – Mycelial growth inhibition rate;

[0084] D0—Correlation diameter of the blank control group;

[0085] D t — Diameter of colonies grown after chemical treatment.

[0086] Experimental statistics: Data were processed using probability value analysis. The DPS statistical analysis system was used to analyze the data and determine the toxicity regression line, R, and EC. 50 Value (95% confidence interval), EC 90 Values ​​(95% confidence interval) are used to evaluate the activity of the test reagent on the biological sample.

[0087] The co-toxicity coefficient (CTC value) of the mixture is calculated using the following formula:

[0088]

[0089] In the formula:

[0090] ATI – Actual Measured Toxicity Index of Mixtures;

[0091] S – EC of standard bactericides 50 The unit is milligrams per liter (mg / L);

[0092] M – EC of the mixture 50 The unit is milligrams per liter (mg / L).

[0093] TTI = TI A *P A +TI B *P B

[0094] In the formula:

[0095] TTI – Theoretical Toxicity Index of Mixtures;

[0096] TI A —A. Toxicity index of drug A;

[0097] P A —Percentage content of drug A in the mixture, expressed as percentage (%);

[0098] TI B —Toxicity index of drug B;

[0099] P B —Percentage content of agent B in the mixture, expressed as percentage (%).

[0100]

[0101] In the formula:

[0102] CTC – Cotoxicity Coefficient;

[0103] ATI – Actual Measured Toxicity Index of Mixtures;

[0104] TTI – Theoretical Toxicity Index of Mixtures.

[0105] 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.

[0106] The test results are shown in the table below:

[0107] Indoor activity tests showed (see Table 2) that the mixture of benzimidazole and pyraclostrobin was effective against EC of citrus anthracnose. 50 The concentrations of pyraclostrobin and benzimidoxam were 0.812 mg / L and 0.145 mg / L, respectively, indicating that the citrus anthracnose pathogen was relatively sensitive to pyraclostrobin. The co-toxicity coefficients of benzimidoxam and pyraclostrobin at ratios of 1:30–25:1 were all greater than 80, showing an additive or synergistic effect in controlling citrus anthracnose. The co-toxicity coefficients of benzimidoxam and pyraclostrobin at ratios of 1:10–14:1 were all greater than 120, showing a synergistic effect in controlling citrus anthracnose. The highest co-toxicity coefficient (139.91) was achieved when benzimidoxam and pyraclostrobin were mixed at a ratio of 7:2. 50 It is 0.287 mg / L.

[0108] Table 2 Results of indoor combined effect test of benzimidazole and pyraclostrobin on citrus anthracnose.

[0109]

[0110] Specific preparation example:

[0111] Preparation Example 1:

[0112] 20% Benzoflufenicol·Pyraclostrobin Suspension Concentrate (15:5)

[0113] Formula: 15% benzo[a]fluconazole, 5% pyraclostrobin, 0.5% sodium dodecyl sulfonate, 1.2% sodium lignosulfonate, 2.5% polyether, 2.5% phenethylphenol polyether phosphate, 1.5% Gelbert alcohol polyoxyethylene ether (XP-70), 1% magnesium aluminum silicate, 0.1% xanthan gum, 5% ethylene glycol, 0.02% benzo[a]isothiazolinone, 0.4% dimethyl silicone oil, deionized water to make up;

[0114] Preparation method: Add benzimidazole and adjuvants (excluding preservatives and thickeners) to a feeding vessel, and start shearing to dissolve all adjuvants. Add pyraclostrobin under high-shear stirring, and after shearing evenly, perform sand milling. After sand milling, transfer to a homogenizing vessel, add preservatives and thickeners, add the remaining water to make up the balance, and then shear and homogenize to obtain the corresponding product.

[0115] Preparation Example 2:

[0116] 32% Benzoflufenicol·Pyraclostrobin Suspension Concentrate (20:12)

[0117] Formula: 20% benzo[a]fluconazole, 12% pyraclostrobin, 2.5% polycarboxylate, 3% phenethylphenol polyether phosphate, 2% isotridecyl alcohol polyoxyethylene ether, 1% magnesium aluminum silicate, 0.2% xanthan gum, 5% propylene glycol, 0.1% benzoic acid, 0.35% dimethyl silicone oil, deionized water to make up;

[0118] Preparation method: Same as in preparation example 1.

[0119] Preparation Example 3:

[0120] 50% Benzoflufenicol·Pyraclostrobin Water Dispersible Granules (36:14)

[0121] Formula: 36% benzyl sulfonate, 14% pyraclostrobin, 5% lignin sulfonate, 7.5% sodium dodecylbenzene sulfonate, 8% aluminum chloride, and precipitated silica as a filler;

[0122] Preparation method: According to the formulation ratio in the example, the active ingredients benzo[i]fluconazole and pyraclostrobin are added to the carrier, and surfactants and other functional adjuvants are added to it. 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 the water-dispersible granule product.

[0123] Example 3

[0124] Field efficacy trial of benzovindiflubenzuron combined with pyraclostrobin for cucumber anthracnose

[0125] Experimental basis: The experiment was conducted in accordance with GB / T 17980.112-2004 "Guidelines for Field Efficacy Tests of Pesticides (II) Part 112: Control of Anthracnose in Cucurbits by Fungicides".

[0126] Experimental crop: Cucumber (Jinchun No. 2).

[0127] Experimental site: Open-field cucumber field in Laixi City. The previous crop was cucumber. The soil fertility was moderate. The planting density was 50,000 plants / hm². 2 The plants grow uniformly and evenly.

[0128] Experimental design: The experimental cells were randomly arranged in blocks, with a cell area of ​​30m². 2 Each cell was repeated 4 times.

[0129] Application time: The cucumber plants were evenly sprayed using a Gongnong 16 backpack sprayer, with a pesticide solution volume of 45 kg / mu. The experiment was conducted with the first application in mid-August 2022, followed by a second application at 7-day intervals, for a total of two applications.

[0130] Experimental investigation: The disease baseline was investigated before the first application of the drug, and the disease index was investigated before the second application (i.e., 7 days after the first application) and 11 days after the second application, and the control effect was calculated. A total of 3 investigations were conducted.

[0131] Survey method: 5 sampling points per plot, 3 plants at each point, and 5-10 leaves on each plant from top to bottom.

[0132] Classification shall be carried out according to the following classification method:

[0133] Grade 0: No lesions;

[0134] Grade 1: The lesion area accounts for less than 5% of the total leaf area;

[0135] Grade 3: The lesion area accounts for 6% to 10% of the total leaf area;

[0136] Level 5: The lesion area accounts for 11% to 25% of the total leaf area;

[0137] Level 7: Lesions cover 26% to 50% of the total leaf area;

[0138] Level 7: The lesion area accounts for more than 51% of the total leaf area;

[0139] Methods for calculating drug efficacy:

[0140]

[0141]

[0142] During the experiment, cucumbers in each treatment plot grew well, and none of the pesticides at the tested concentrations caused phytotoxicity to cucumber plants or other non-target organisms.

[0143] The results of the field efficacy trials are shown in the table below:

[0144] Table 3. Results of field efficacy trials of benzo[a]fluconazole and pyraclostrobin combined for cucumber anthracnose.

[0145]

[0146]

[0147] Field efficacy results (see Table 3): Under the same environmental conditions, the overall control efficacy of different treatments for cucumber anthracnose ranged from 69.48% to 83.21% 7 days after the first application. Spraying with 50% benzalkonium chloride + pyraclostrobin wettable powder at 80 g / hm² was most effective. 2 100g / hm 2 120g / hm 2 The control effects of the three different dosage treatments on cucumber anthracnose varied, with rates of 81.09%, 82.72%, and 83.21%, respectively, which were superior to the control agents (45% benzimidazole·pyraclostrobin water-dispersible granules (30+15), 9% benzimidazole emulsifiable concentrate, and 25% pyraclostrobin suspension).

[0148] Eleven days after the second application, the overall control efficacy of each treatment in the field ranged from 71.79% to 85.63%. Spraying with 80 g / hm² of 50% benzalkonium chloride + pyraclostrobin wettable powder was recommended. 2 100g / hm 2 120g / hm 2 The control efficacy against cucumber anthracnose was 83.15%, 84.21%, and 85.63% after three different dose treatments, respectively.

[0149] Through indoor toxicity testing and field trials, the pesticide composition of benzo[a]flufenicol and pyraclostrobin described in this invention showed good control effect against plant anthracnose.

[0150] The pesticide compositions or formulations obtained by this invention exhibit significant preventive effects, demonstrating superior efficacy compared to single agents in delaying the development of resistance and extending the duration of action. Furthermore, no phytotoxicity was observed in the experiments with the compounded pesticides, indicating that the enhanced synergistic bactericidal effect of the resulting pesticide compositions or formulations can reduce production and usage costs while ensuring crop safety.

[0151] 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, 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. The use of a fungicide composition containing benzimidazole and pyraclostrobin for the control of anthracnose diseases in plants, characterized in that: The fungicidal composition contains active ingredient A, benzimidazole, and active ingredient B, pyraclostrobin, wherein the mass ratio of benzimidazole to pyraclostrobin is 1:10 to 14:1, and the plant anthracnose is caused by *Colletotrichum spp.* (Cucurbitaceae). Colletotrichum orbiculare Cucumber anthracnose caused by collodion ( ) or by colloidal anthracnose ( ) Colletotrichum gloeosporioides Citrus anthracnose caused by (Penz.) Penz.et Sacc.

2. The use according to claim 1, characterized in that: The mass ratio of the active ingredient benzo[a]fluconazole to pyraclostrobin is 1:10, 1:5, 1:3, 3:1, 7:2, or 14:

1.

3. The use according to claim 1, characterized in that: Based on a total weight of 100 wt% of the bactericidal composition, the sum of the contents of benzylfloxacin and pyraclostrobin in the bactericidal composition is 5-90 wt%.

4. The use according to claim 3, characterized in that: Based on a total weight of 100 wt% of the bactericidal composition, the sum of the contents of benzo[i]flufenicol and pyraclostrobin in the bactericidal composition is 5-80 wt%.

5. The use according to claim 1, characterized in that: The bactericidal composition is formulated as any one of emulsifiable concentrate, water emulsion, microemulsion, suspension concentrate, wettable powder, or water-dispersible granules.

Images