Bactericidal composition containing isofetamid and benziothiazolinone

Abstract

The invention belongs to the technical field of pesticides, and particularly relates to a bactericidal composition containing isofetamid and benziothiazolinone, the active ingredients of the bactericidal composition comprise (A) isofetamid and (B) benziothiazolinone, the weight ratio of the component (A) to the component (B) is 15: 1-1: 15, and the total weight of isofetamid and benziothiazolinone accounts for 5-90% of the weight of the composition. In combination with actual application requirements, the bactericidal composition can be prepared into any one of an emulsion in water, a microemulsion, a suspending agent, water dispersible granules and wettable powder, and is used for preventing and treating common fungal and bacterial diseases in agricultural production.

Description

Technical Field

[0001] The invention relates to a new field of pesticide technology, and in particular to a fungicide composition containing isoflurane and thiophanate-methyl, which is used for preventing and controlling crop diseases. Background Art

[0002] Isofetamid belongs to the class of phenyloxyethylthiophenamide compounds and is a SDHI fungicide with a thiophenamide structure. Its mechanism of action is to act on the mitochondrial respiration of pathogens, completely or partially occupy the site of the substrate ubiquinone, inhibit the transfer of electrons from succinate to ubiquinone, thereby blocking the energy metabolism of pathogens, inhibiting the growth of pathogens, and causing their death, thereby achieving the purpose of preventing and controlling plant diseases. Isofetamid is broad-spectrum and highly effective, with protective, therapeutic, penetrating (or local systemic) effects, and can effectively control Ascomycetes and Deuteromycetes fungi, and prevent and control many foliar and soil-borne diseases. It is mainly used for fruit trees, vegetables, grapes, rapeseed, dwarf berries and berries, etc., to prevent and control diseases caused by pathogens such as Sclerotiniaspp., Botrytis spp., Monilia spp., such as gray mold, sclerotinia, powdery mildew, anthracnose, brown spot, apple scab, etc.

[0003] Benziothiazolinone belongs to the class of thiazole heterocyclic compounds and is a new type of highly effective, low-toxic, broad-spectrum fungicide. Its mechanism of action is: by combining with anions on the surface of bacterial and fungal cell membranes or reacting with sulfhydryl groups, it destroys the synthesis system of proteins and cell membranes, thereby effectively inhibiting the reproduction of bacteria and fungi, interfering with the metabolism of pathogenic bacteria cells, causing physiological disorders and leading to the death of pathogens. This compound is mainly used to prevent and control common diseases such as rot, canker, anthrax, black spot, leaf spot, downy mildew, etc. that often occur on crops.

[0004] JP2015044791A discloses an agricultural chemical composition containing as an active ingredient a component selected from 1,2-benzisothiazole derivatives, and one or more other compounds such as clothianidin, imidacloprid, dinotefuran, flubendiamide, and the like.

[0005] With the widespread occurrence of fungal and bacterial diseases in agriculture around the world, the use of pesticides has increased year by year, and the resistance of pathogens has become increasingly serious, and the number of resistant pathogens has continued to increase. At the same time, the high-intensity use of pesticides and non-standard use of pesticides have also led to excessive pesticide residues in agricultural products, resulting in a series of food security problems, and also greatly increased the cost of pesticides for farmers, which is not conducive to the green, healthy and sustainable development of agriculture. Summary of the invention

[0006] In order to solve the problems existing in the prior art, the present invention has found through indoor and field efficacy tests that both isoprene and thiophanate-methyl have good control effects on fungal and bacterial diseases within a certain ratio range, and the synergistic effect of the two is obvious. This combination greatly reduces the amount of pesticides used to a certain extent, and also reduces the cost of pesticides for farmers and the risk of disease resistance, which has positive significance for the sustainable development of agriculture. Based on this, the present invention is completed.

[0007] Therefore, the object of the present invention is to provide a fungicide composition with significant synergistic effect, broad bactericidal spectrum, long lasting effect, low toxicity, low residue, low drug cost, safe and harmless to crops, and not easy to produce resistance.

[0008] The technical scheme adopted by the present invention is: a fungicide composition containing isoflavone and thiophanate-methyl, wherein the active ingredients are isoflavone and thiophanate-methyl, and the weight ratio of the two is 15:1 to 1:15.

[0009] In a preferred embodiment of the present invention, the weight ratio of isoprofenamide to thiophanate-methyl is preferably 10:1 to 1:10, for example, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10; more preferably 5:1 to 1:5.

[0010] In the fungicidal composition of the present invention, the total weight percentage of isoprenaline and thiophanate-methyl in the composition is 5% to 90%, preferably 10% to 60%, and the rest are agricultural adjuvants allowed to be added and used in agricultural production.

[0011] In a preferred embodiment of the present invention, the fungicidal composition containing isoprene and thiophanate-methyl can be prepared into a suitable agricultural formulation by adding suitable pesticide adjuvants according to methods well known to those skilled in the art, including but not limited to: aqueous emulsions, microemulsions, suspensions, water dispersible granules, and wettable powders.

[0012] In a preferred embodiment of the present invention, the agricultural formulation prepared from the pesticide composition is preferably a suspension concentrate.

[0013] In the preparation process of the fungicide composition dosage form of the present invention, the pesticide adjuvants used include: dispersants, wetting agents, emulsifiers, thickeners, preservatives, defoaming agents, antifreeze agents, etc. The specific adjuvant varieties are also known. The adjuvants help the active ingredients to be stable in the formulation system and exert their efficacy, and they are all common and permitted in pesticide preparations.

[0014] For example: Auxiliary ingredients in suspensions include but are not limited to:

[0015] The dispersant can be selected from one or a mixture of alkylbenzene sulfonate, alkylnaphthalenesulfonic acid formaldehyde condensate, cresol sulfonic acid / naphthol sulfonic acid formaldehyde condensate, N-methyl fatty sulfonic acid formaldehyde condensate, lignin sulfonate, alkylphenol polyoxyethylene ether formaldehyde condensate sulfate, fatty alcohol polyoxyethylene ether sulfate, alkyl phosphate, fatty alcohol polyoxyethylene ether phosphate, alkyl (aryl) phenol polyoxyethylene ether phosphate, etc.;

[0016] The wetting agent can be selected from one or a mixture of nonylphenol polyoxyethylene ether, fatty alcohol polyoxyethylene ether, alkanolamide polyoxyethylene ether, phosphate or sulfate ester salt, alkyl polyoxyethylene ether succinate sulfonate, alkylbenzene sulfonate, alkylnaphthalene sulfonate, lignin sulfonate, polycarboxylate, etc.;

[0017] The emulsifier can be selected from one or a mixture of polyoxyethylene-polyoxypropylene block copolymer, emulsifier BP, styrylphenyl polyoxyethylene ether, fatty alcohol polyoxyethylene ether, alkylphenol polyoxyethylene ether, alkylphenol polyoxyethylene polyoxypropylene ether, etc.;

[0018] The thickener may be one or a mixture of gum arabic, xanthan gum, carboxymethyl cellulose, polyvinyl alcohol, gaseous silicon dioxide, magnesium aluminum silicate, carboxyethyl cellulose, carboxypropyl cellulose, sodium acrylate, polyvinyl pyrrolidone, bentonite, white carbon black, etc.;

[0019] The preservative may be selected from one or a mixture of kason, formaldehyde, benzoic acid, sodium benzoate, sodium sorbate, isothiazolinone;

[0020] The defoamer can be selected from one or a mixture of organic silicon, silicone compounds, C10-20 saturated fatty acid compounds, C8-10 fatty alcohols, hexanol, butanol;

[0021] The antifreeze agent can be selected from one or a mixture of glycerol-ether diglycol, methyl propylene diglycol, ethylene glycol, propylene glycol, glycerol, and urea;

[0022] The water is deionized water.

[0023] Another aspect of the present invention relates to a method for using the above-mentioned fungicidal composition containing isoprene and thiophanate-methyl to prevent and control agricultural diseases, specifically, contacting the fungicidal composition with crops or agricultural pathogens to inhibit the normal growth of pathogens until they die to achieve the purpose of preventing and controlling agricultural diseases. There is no particular limitation on the specific method of application, and the application can be done in a manner familiar to those skilled in the art, such as by spraying. There is no particular limitation on the frequency and amount of application, and they can vary according to the type of crop, the growth period, the occurrence of the disease, and the weather. The occurrence of the disease can be effectively prevented and controlled by using an appropriate dosage.

[0024] In the method of the present invention, the crop is not particularly limited, and can be used for known conventional crops, preferably lettuce, peanut, strawberry, and tomato. The agricultural disease is not particularly limited, for example, it can be used to prevent and control various agricultural diseases known in the art, preferably for preventing and controlling lettuce sclerotinia, peanut brown spot, strawberry gray mold, and tomato gray mold.

[0025] Another aspect of the present invention relates to the use of the fungicidal composition containing isoprenaline and thiophanate-methyl for preventing and controlling agricultural diseases. In the use of the present invention, the application method and application amount are as described above.

[0026] In the use of the present invention, the agricultural diseases are not particularly limited, and can be used to control various agricultural diseases known in the art, preferably to control lettuce sclerotinia, peanut brown spot, strawberry gray mold, tomato gray mold, etc. The fungicidal composition containing isoprenaline and thiophanate-methyl of the present invention exhibits excellent synergistic effects in controlling the above agricultural diseases.

[0027] The present invention has the following advantages:

[0028] (1) The active ingredients of the fungicidal composition provided by the present invention are isoprofenamide and thiophanate-methyl, which show obvious synergistic effect within a certain ratio range. The control effect of the composition is significantly better than that of a single agent, which greatly reduces the application amount of pesticides, reduces the impact of pesticides on the environment, and saves the cost of drug use.

[0029] (2) The active ingredients of the fungicidal composition provided by the present invention have different mechanisms of action, with more action sites, which delays or overcomes the development of drug resistance in pathogens, thereby effectively extending the service life of the product, improving its utilization rate, and facilitating the comprehensive prevention and control of diseases.

[0030] (3) The fungicidal composition of the present invention has a good control effect on diseases such as lettuce sclerotinia rot, peanut brown spot disease, strawberry gray mold and tomato gray mold, and is safe to use without adverse effects, meeting the requirements of national food security. DETAILED DESCRIPTION

[0031] The technical scheme and technical effects of the present invention are further described below in conjunction with specific embodiments, but the implementation manner of the present invention is not limited to the scope described in the embodiments.

[0032] Unless otherwise specified, the percentages described in the present invention are all weight percentages. The instruments, reagents, materials, etc. involved in the following implementation cases, unless otherwise specified, are all conventional instruments, reagents, materials, etc. already available in the prior art and can be obtained through regular commercial channels.

[0033] The test methods, detection methods, etc. involved in the following implementation cases, unless otherwise specified, are all conventional test methods, detection methods, etc. in the prior art. Active ingredients are all calculated as effective ingredients.

[0034] This experiment used a combination of indoor toxicity determination and field test. First, the indoor toxicity determination was used to screen out the appropriate ratio, and then the field test was carried out on this basis.

[0035] 1. Preparation Examples

[0036] Example 1 (15+1)% isoprofenamide·thiophanate-methyl suspension

[0037]

[0038] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0039] Example 2 (20+2)% Isoprothiocarb·Thiophanate-methyl suspension

[0040]

[0041] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0042] Example 3 (15+3)% Iprodione-Thiophanate-Methyl-1,4-Dimethomorph Suspension Concentrate

[0043]

[0044] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0045] Example 4 (5+5)% Iprothiocarb·Thiophanate-methyl suspension

[0046]

[0047] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0048] Example 5 (3+15)% Iprothiocarb·Thiophanate-methyl suspension

[0049]

[0050] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0051] Example 6 (2+20)% Iprodione-Thiophanate-Methyl-1,4-D-Isopropylamine Suspension Concentrate

[0052]

[0053] The above components are mixed in proportion, ground or sheared at high speed for a certain period of time, and then the zirconium beads are filtered to obtain a suspension.

[0054] Example 7 (1+15)% Iprothiocarb·Thiophanate-methyl suspension

[0055]

[0056] The above components are mixed in proportion and then ground or filtered at high speed for a certain period of time to obtain a suspension.

[0057] 1. Indoor bioassay activity determination

[0058] Example 1: Screening of compound ratios of isoprofenamide and thiophanate-methyl against lettuce sclerotinia sclerotiorum and determination of co-toxicity coefficient

[0059] 1.1 Test strains

[0060] The pathogen of lettuce sclerotinia disease was Sclerotinia sclerotiorum (Libert) de Bery of the Sclerotiniaceae family of the subphylum Ascomycetes, which was provided by the Fungal Disease Research Laboratory of the College of Agriculture, Zhejiang A&F University.

[0061] 1.2 Test culture medium

[0062] Potato dextrose agar (PDA) medium was used: 200 g potato, 20 g glucose, 20 g agar, and deionized water was added to 1000 ml.

[0063] 1.3 Test agents

[0064] 95% isoprenaline technical (Ishihara Sangyo Co., Ltd., Japan), 95% thiophanate-methyl technical (Shaanxi Xida Huate Technology Industry Co., Ltd.). 95% isoprenaline technical and 95% thiophanate-methyl technical were dissolved in DMF to prepare mother liquors, which were diluted with sterile water containing 2% Tween 80 when used.

[0065] 1.4 Indoor toxicity test of isoflurane and thiophanate-methyl against lettuce sclerotinia rot

[0066] Test method: refer to "Agricultural Industry Standard of the People's Republic of China NY / T1156.2-006", mycelium growth rate method.

[0067] Use a pipette to draw 1 ml of the prepared drugs in different ratios and add them to a conical flask containing 44 ml of PDA culture medium that has been sterilized and cooled to 40℃~50℃. After fully mixing, pour the mixture evenly into 3 culture dishes to prepare drug-containing plates of corresponding concentrations. Set the drug-free culture medium as a blank control (using sterile water containing 2% Tween 80 instead of the drug). The lettuce sclerotinia pathogen that has been cultured at 25℃ for 2~3 days is punched along the edge of the colony with a sterile puncher with a diameter of 5mm under sterile conditions to obtain a bacterial cake. The bacterial cake is then inoculated with an inoculator in the center of the drug-containing PDA plate, with the mycelium side facing down. The plate is covered with a lid and turned upside down. Each treatment is repeated 3 times and then cultured in an artificial incubator at 25℃ for 3 days. The colony diameter is determined by the cross method, and the average value of the 3 replicates of each concentration is taken. The EC values ​​of different ratios of drugs against the pathogen are then calculated. 50 , and the co-toxicity coefficient method is used to calculate the co-toxicity coefficient (CTC) to determine the synergistic effect of the mixture. The specific calculation method is as follows:

[0068] Take a single agent in the mixture as the standard agent (usually EC 50 The lower one), calculate:

[0069] Measured Toxicity Index (ATI) = (Standard Agent EC 50 / Test Drug EC 50 )×100

[0070] Theoretical toxicity index (TTI) = toxicity index of agent A × percentage of A in the mixture + toxicity index of agent B × percentage of B in the mixture

[0071] Co-toxicity coefficient (CTC) = [Actual toxicity index of mixture (ATI) / Theoretical toxicity index of mixture (TTI) × 100

[0072] Co-toxicity coefficient classification: When the co-toxicity coefficient (CTC) is ≥120, the mixture exhibits a synergistic effect; when the co-toxicity coefficient (CTC) is ≤80, it exhibits an antagonistic effect; and when the co-toxicity coefficient (CTC) is between 80 and 120, it exhibits an additive effect.

[0073] Table 1 Indoor joint toxicity test of isoflurane and thiophanate-methyl against lettuce sclerotinia sclerotiorum

[0074] Drug name and ratio Ratio <![CDATA[EC 50 (mg / L)]]> ATI TTI CTC Iprodione(A) —— 1.75 100.00 Thiophanate(B) —— 26.37 6.64 A: B 15:1 1.28 136.72 94.16 145.19 A: B 10:1 1.15 152.17 91.51 166.29 A: B 5:1 1.14 153.51 84.44 181.80 A: B 1:1 2.39 73.22 53.32 137.33 A: B 1:5 4.61 37.96 22.20 171.02 A: B 1:10 7.58 23.09 15.12 152.65 A: B 1:15 10.25 17.07 12.47 136.90

[0075] As shown in Table 1, the EC values ​​of isoprofenamide and thiophanate-methyl for controlling lettuce sclerotinia sclerotiorum 50The concentrations of 1.75 mg / L and 26.37 mg / L were 1.75 mg / L and 26.37 mg / L, respectively. When the ratio of the two was 15:1 to 1:15, they showed good synergistic effects on lettuce sclerotinia, and their co-toxicity coefficients (CTC) were all greater than 120. Especially when the ratio was 5:1, the synergistic effect was more significant, and the co-toxicity coefficient (CTC) was 181.80. This shows that the combination of isoprofenamide and thiophanate-methyl is reasonable and feasible.

[0076] Example 2: Screening of compound ratios of isoprenaline and thiophanate-methyl against peanut brown spot disease and determination of co-toxicity coefficient

[0077] 2.1 Test pathogens

[0078] The pathogen of peanut brown spot disease was Cercospora arachidicola Hori, which was collected from naturally diseased peanut leaves in the field in Zhengyang, Henan Province. Conidia of peanut brown spot disease were picked from the spots on peanut leaves with typical symptoms and placed in sterile water for shaking for 3 minutes. The conidia suspension was spread on a water agar plate, and then a single spore was picked with an inoculation needle and placed on a potato dextrose (PDA) medium, and then cultured in the dark in a 26°C incubator. The pathogen was identified by observing the colony and spore morphology combined with ITS sequence comparison and then transferred to the PDA plate. After the hyphae grew, the colony was picked with an inoculation needle and transferred to the PDA slant and stored at 4°C for later use.

[0079] 2.2 Test culture medium

[0080] Potato dextrose agar (PDA) medium was used: 200 g potato, 20 g glucose, 20 g agar, and deionized water was added to 1000 ml.

[0081] 2.3 Tested fungicides

[0082] 95% isoprenaline technical (Ishihara Sangyo Co., Ltd., Japan), 95% thiophanate-methyl technical (Shaanxi Xida Huate Technology Industry Co., Ltd.). 95% isoprenaline technical and 95% thiophanate-methyl technical were dissolved in DMF to prepare mother liquors, which were diluted with sterile water containing 2% Tween 80 when used.

[0083] 2.4 Indoor toxicity test of isoflurane and thiophanate-methyl against peanut brown spot disease

[0084] Test method: refer to "Agricultural Industry Standard of the People's Republic of China NY / T1156.2-006", mycelium growth rate method.

[0085] Use a pipette to take 1 ml of the prepared drugs in different ratios and add them to a conical flask containing 44 ml of PDA culture medium that has been sterilized and cooled to 40℃-50℃. After fully mixing, pour the mixture evenly into 3 culture dishes to prepare drug-containing plates of corresponding concentrations. Set the drug-free culture medium as a blank control (use sterile water containing 2% Tween 80 instead of the drug). Under sterile conditions, use a sterile puncher with a diameter of 5 mm to punch holes along the edge of the colony to take out the bacterial cake. Then, use an inoculator to inoculate the punched bacteria into the center of the drug-containing culture medium with the mycelium facing down. Cover the dish with a lid, repeat 3 times for each treatment, and finally place it in a constant temperature box at 28℃ for culture. When the blank peanut brown spot pathogen is about to fill the culture dish, use the cross method to measure the colony diameter in each culture dish, and take the average of the 3 repetitions of each concentration. Then calculate the EC values ​​of different ratios of drugs against pathogens. 50 , and the co-toxicity coefficient method is used to calculate the co-toxicity coefficient (CTC) to determine the synergistic effect of the mixture. The specific calculation method is the same as above.

[0086] Table 2 Indoor joint toxicity test of isoflurane and thiophanate-methyl against peanut brown spot disease

[0087]

[0088]

[0089] As shown in Table 2, the EC values ​​of isoprofenamide and thiophanate-methyl for controlling peanut brown spot disease 50 When the ratio of the two is 15:1 to 1:15, they both show good synergistic effect on peanut brown spot disease, and their co-toxicity coefficient (CTC) is greater than 120. Especially when the ratio is 5:1, the synergistic effect is more significant, and the co-toxicity coefficient (CTC) is 173.22. It can be seen that the combination of isoprenaline and thiophanate-methyl is reasonable and feasible.

[0090] Example 3: Screening of compound ratios of isoprothiocarb and thiophanate-methyl against strawberry gray mold and determination of co-toxicity coefficient

[0091] 3.1 Test pathogens

[0092] The strawberry gray mold pathogen was Botrytis cinerea Pers, which belongs to the subphylum Ascomycota and was provided by the Plant Pathology Laboratory, Department of Plant Protection, Guizhou University.

[0093] 3.2 Test culture medium

[0094] Potato dextrose agar (PDA) culture medium was used: 200 g potato, 20 g glucose, 20 g agar, and deionized water was added to 1000 ml.

[0095] 3.3 Tested fungicides

[0096] 95% isoprenaline technical (Ishihara Sangyo Co., Ltd., Japan), 95% thiophanate-methyl technical (Shaanxi Xida Huate Technology Industry Co., Ltd.). 95% isoprenaline technical and 95% thiophanate-methyl technical were dissolved in DMF to prepare mother liquors, which were diluted with sterile water containing 2% Tween 80 when used.

[0097] 3.4 Toxicity determination of isoprofenamide and thiophanate-methyl against strawberry gray mold

[0098] Test method: refer to "Agricultural Industry Standard of the People's Republic of China NY / T1156.2-006", mycelium growth rate method.

[0099] Use a pipette to draw 1 ml of the prepared drugs in different ratios and add them to a sterilized conical flask containing 44 ml of PDA culture medium cooled to 40℃~50℃. After fully mixing, pour them evenly into 3 culture dishes to make drug-containing plates of corresponding concentrations. Set the drug-free culture medium as a blank control (use sterile water containing 2% Tween 80 instead of the drug). Under sterile conditions, use a sterile puncher to punch a 5mm bacterial cake from the edge of the strawberry gray mold pathogen cultured on the PDA culture medium for 3 days. Then use an inoculator to inoculate the beaten bacterial cake into the center of the drug-containing culture medium with the mycelium facing down and cover the dish. Repeat 3 times for each treatment and then culture in an artificial incubator at 25℃. Check the mycelium growth 5 days after inoculation and use the cross method to measure the colony diameter in each culture dish. Take the average of the 3 replicates of each concentration. Then calculate the EC values ​​of different ratios of drugs against pathogens. 50 , and the co-toxicity coefficient method is used to calculate the co-toxicity coefficient (CTC) to determine the synergistic effect of the mixture. The specific calculation method is the same as above.

[0100] Table 3 Indoor joint toxicity test of isoprofenamide and thiophanate-methyl against strawberry gray mold

[0101] Drug name and ratio Ratio EC50(mg / L) ATI TTI CTC Iprodione(A) —— 2.67 100.00 Thiophanate(B) —— 18.36 14.54 A: B 15:1 1.92 139.06 94.66 146.91 A: B 10:1 1.75 152.57 92.23 165.42 A: B 5:1 1.66 160.84 85.76 187.56 A: B 1:1 3.07 86.97 57.27 151.86 A: B 1:5 5.29 50.47 28.79 175.34 A: B 1:10 8.18 32.64 22.31 146.30 A: B 1:15 9.87 27.05 19.88 136.05

[0102] As shown in Table 3, the EC values ​​of isoprofenamide and thiophanate-methyl for controlling strawberry gray mold 50 When the ratio of the two is 15:1 to 1:15, they both show good synergistic effect on strawberry gray mold, and their co-toxicity coefficient (CTC) is greater than 120. Especially when the ratio is 5:1, the synergistic effect is more significant, and the co-toxicity coefficient (CTC) is 187.56. This shows that the combination of isoprenaline and thiophanate-methyl is reasonable and feasible.

[0103] Example 4: Screening of compound ratios of isoprofenamide and thiophanate-methyl against tomato gray mold and determination of co-toxicity coefficient

[0104] 4.1 Test pathogens

[0105] The tomato gray mold pathogen was Botrytis cinerea Pers.Fr, which belongs to the genus Botrytis of the subphylum Ascomycota and was provided by the Plant Protection Pathology Laboratory of Nanjing Agricultural University and stored at 4°C.

[0106] 4.2 Test culture medium

[0107] Potato sucrose agar (PSA) culture medium was used: 200 g potato, 20 g sucrose, 20 g agar, and deionized water was added to make up to 1000 ml.

[0108] 4.3 Test fungicides

[0109] 95% isoprenaline technical (Ishihara Sangyo Co., Ltd., Japan), 95% thiophanate-methyl technical (Shaanxi Xida Huate Technology Industry Co., Ltd.). 95% isoprenaline technical and 95% thiophanate-methyl technical were dissolved in DMF to prepare mother liquors, which were diluted with sterile water containing 2% Tween 80 when used.

[0110] 4.4 Toxicity determination of isoprofenamide and thiophanate-methyl against tomato gray mold

[0111] Test method: refer to "Agricultural Industry Standard of the People's Republic of China NY / T1156.2-006", mycelium growth rate method.

[0112] Use a pipette to draw 1 ml of the prepared drugs in different ratios and add them to a conical flask containing 44 ml of PSA culture medium that has been sterilized and cooled to 40℃~50℃. After fully mixing, pour the mixture evenly into 3 culture dishes to prepare drug-containing plates of corresponding concentrations. Set the medium without drugs as a blank control (use sterile water containing 2% Tween 80 instead of drugs). Under sterile conditions, use a sterile puncher to punch a 5 mm bacterial cake from the edge of the tomato gray mold pathogen cultured at 25℃ for 4 days on the PSA culture medium plate. Then use an inoculator to inoculate the beaten bacterial cake into the center of the drug-containing culture medium, with the mycelium facing down. Cover the dish with a lid, repeat 3 times for each treatment, and then culture in an artificial incubator at 25℃. Check the mycelium growth 5 days after inoculation and use the cross method to measure the colony diameter in each culture dish. Take the average of the 3 replicates of each concentration. Then calculate the EC values ​​of different ratios of drugs against pathogens. 50 , and the co-toxicity coefficient method is used to calculate the co-toxicity coefficient (CTC) to determine the synergistic effect of the mixture. The specific calculation method is the same as above.

[0113] Table 4 Indoor joint toxicity test of isoflurane and thiophanate-methyl against tomato gray mold

[0114] Drug name and ratio Ratio EC50(mg / L) ATI TTI CTC Iprodione(A) —— 1.32 100.00 Thiophanate(B) —— 15.67 8.42 A: B 15:1 1.01 130.69 94.28 138.63 A: B 10:1 0.93 141.94 91.67 154.82 A: B 5:1 0.88 150.00 84.74 177.02 A: B 1:1 1.64 80.49 54.21 148.47 A: B 1:5 3.39 38.94 23.69 164.39 A: B 1:10 5.21 25.34 16.75 151.27 A: B 1:15 7.34 17.98 14.15 127.12

[0115] As shown in Table 4, the EC values ​​of isoprofenamide and thiophanate-methyl for controlling tomato gray mold 50 When the ratio of the two is 15:1 to 1:15, they both show good synergistic effect on tomato gray mold, and their co-toxicity coefficient (CTC) is greater than 120. Especially when the ratio is 5:1, the synergistic effect is more significant, and the co-toxicity coefficient (CTC) is 177.02. It can be seen that the combination of isoprenaline and thiophanate-methyl is reasonable and feasible.

[0116] 3. Field efficacy test

[0117] Example 1: Field efficacy test of the combination of isoprenaline and thiophanate-methyl against lettuce sclerotinia rot

[0118] Selection of test site: The test site is located in Jinhua, Zhejiang, and is a transplanted lettuce field. The test field has flat terrain, medium fertility, pH value of 6.5, and organic matter content of 4.5%. The growth and planting density of lettuce in the test field are uniform and consistent.

[0119] Experimental treatment: The experimental agent 18% isoprene·thiopyrad suspension (Formulation Example 3) was set at 750, 1000, and 1500 times of solution for three treatments, 5% thiopyrad suspension was set at 1000 times, 15% isoprene suspension was set at 1200 times and clear water control for a total of 6 treatments, 4 replicates, a total of 24 plots, each plot area was 25m 2 .

[0120] Application: After the lettuce was transplanted and survived, the first application was made at the rosette stage (August 10), the second application was made on August 17, and the third application was made on August 24. 2 Add 60kg of water and use a backpack manual sprayer to spray evenly, focusing on the middle and lower parts of the plant.

[0121] Investigation and recording method: 12 days after the third application of the drug, the disease of lettuce sclerotinia was stable, and the efficacy of the drug was investigated. Samples were taken at 5 points in the middle of each plot, and 5 plants were investigated at each point. The number of diseased plants was counted, the disease index and control effect were calculated, and the difference was significantly determined. The grading standards for lettuce sclerotinia are as follows:

[0122] Level 0 – no disease;

[0123] Level 1: There are a few lesions at the stem base or leaves;

[0124] Level 2: Nearly one-third of the stem base or outer leaves are rotten;

[0125] Level 3: More than one-third of the plant is rotten.

[0126] Efficacy calculation method: Calculate according to the national standard "Guidelines for Field Efficacy Tests of Pesticides (I)" method; calculate the disease index and control effect. Use the Duncan's New Multiple Range (DMRT) method to conduct a significance analysis of the control effect. During the test, the safety of the pesticide on lettuce was also investigated.

[0127] Disease index = 100 × ∑ (number of diseased leaves at each level × representative value of each level) / (total number of leaves surveyed × highest representative value)

[0128] Control effect (%) = (CK1-P1) / CK1×100

[0129] P1 disease index: disease index of the treated area after prevention and treatment;

[0130] CK1 disease index: disease index of the control area after prevention and treatment;

[0131] The results are shown in Table 5:

[0132] Table 5 Statistics of field control of lettuce sclerotinia by each treatment agent

[0133]

[0134]

[0135] Note: Different lowercase letters in the same column represent significant differences among treatments (P<0.05).

[0136] As can be seen from Table 5, when the formulation example 3: 18% isoprene·thiophanate-methyl SC is diluted 750 times, the effect of preventing and controlling lettuce sclerotinia is the best, with a control efficiency of 91.59%. At the same time, when the dilution multiples are 1000 times and 1500 times, the control effects are 86.62% and 79.75%, respectively. Compared with 5% thiophanate-methyl SC and 15% isoprene SC, both have good control effects and a long lasting effect. In addition, by observing the growth of lettuce in the field, it was found that within the normal range of medication, the lettuce grew well and no drug damage was found. Therefore, it is safe to use isoprene and thiophanate-methyl under this ratio for lettuce. At the same time, the agent also has a good preventive effect on the occurrence of soft rot of lettuce in the field.

[0137] Example 2: Field efficacy test of the combination of isoprenaline and thiophanate-methyl against peanut brown spot disease

[0138] The experimental site was selected in the peanut experimental field in Baichi Township, Shangcai County, Zhumadian City, Henan Province. The experimental field was sandy loam, the variety was Huayu 23, which is susceptible to brown spot disease, and the cultivation method was open-field ridge cultivation. The management measures were relatively consistent, the fertilizer and water management was at a medium to high level, and the peanut growth was basically the same. When the pesticide was first applied in the experiment, the peanut brown spot disease was in the early stage of the disease and there was no obvious disease manifestation.

[0139] Experimental treatment: 18% isoprene·thiopyrad suspension (Example 3) was set at 750, 1000, and 1500 times, 5% thiopyrad suspension was set at 1000 times, 15% isoprene suspension was set at 1200 times and a total of 6 treatments of water control, each treatment was repeated 4 times, randomly arranged, a total of 24 plots, each plot area was 15m 2 .

[0140] Application: The test was conducted in June 2024, based on 667m 2 Add 60kg of water for application, and use a backpack manual sprayer to evenly spray the front and back surfaces of the peanut leaves until they are wet. Apply the medicine 3 times in total. The first application time is when peanut brown spot disease occurs sporadically, and then spray once every 7 days, for a total of 3 times. The application time is June 8, 15, and 23.

[0141] Investigation and recording method: Conduct a prevention effect investigation during the stable period of peanut brown spot disease. The specific investigation time is 10 days after the last application of the pesticide. Conduct a disease investigation according to the jump method. In each plot, 5 rows are investigated at intervals, and 10 plants are surveyed in each row. According to the following calculation formula, calculate the disease index and field prevention effect, and conduct a significant difference determination. After the peanuts in each treatment plot are harvested and dried, the pod yield is measured and compared with CK. Calculate the increase in yield and compare the effects of different types of fungicides and dosages on peanut yield. The classification standards for peanut brown spot disease are as follows:

[0142] Grade 1 – no symptoms, 0% leaf area loss;

[0143] Level 3: There are many lesions on the lower leaves, and the leaves fall off obviously. There are a few lesions on the middle leaves, and the leaf area loss rate is 6% to 10%;

[0144] Level 5: All the middle and lower leaves are infected, 50% of the lower leaves fall off, and the leaf area loss rate is 21% to 30%;

[0145] Level 7: Severe lesions on the middle and lower leaves, severe leaf fall, lesions on the upper leaves, and leaf area loss rate of 41% to 60%;

[0146] Level 9: Almost all leaves fall, with only a small number of leaves with diseased spots on the upper part remaining. The leaf area loss rate is 81% to 100%.

[0147] Efficacy calculation method: Calculate according to the national standard "Pesticide Field Efficacy Test Guidelines (I)" method; calculate the disease index and control effect. Use Duncan's new multiple range (DMRT) method to perform significance analysis on the control effect, and investigate the safety of the pesticide on peanuts during the test. (The disease index and control effect are the same as those in field Example 1.)

[0148] The results are shown in Tables 6 and 7:

[0149] Table 6 Statistics of field control of peanut brown spot disease by each treatment agent

[0150]

[0151]

[0152] Table 7 Effects of various treatment agents on peanut yield

[0153]

[0154] Note: Different lowercase letters in the same column represent significant differences among treatments (P<0.05).

[0155] As can be seen from Tables 6 and 7, when the formulation example 3: 18% isoprene·thiopyrad SC is diluted 750 times, the effect of preventing and controlling peanut brown spot disease is the best, with a prevention effect of 87.86%. At the same time, when the dilution multiple is 1000 times and 1500 times, the prevention and control effects are 79.32% and 76.78%, respectively. Compared with 5% thiopyrad SC and 15% isoprene SC, both have good prevention and control effects and a long lasting effect. The maximum increase in yield of peanuts measured by production is 33.28%, and the yield increase effect is significant. In addition, under dilution of 1000 times and 1500 times, there is also a good yield increase effect on peanut brown spot disease. At the same time, the growth of peanuts in the field was observed. Within the normal range of medication, the growth of peanuts was good, and no phytotoxicity was found. It is an ideal agent for preventing and controlling peanut brown spot disease.

[0156] Example 3: Field efficacy test of the combination of isoprenaline and thiophanate-methyl against strawberry gray mold

[0157] Selection of test site: The test was conducted in a greenhouse in Shibuzi Town, Anqiu, Shandong. The strawberry variety was "Hongyan". The soil was clay loam with an organic matter content of 3.75% and a pH of 6.8. The test strawberries were transplanted on September 10, 2022. The planting greenhouse is a three-span greenhouse, each with a width of 6m and a length of 60m. There are 6 beds, 2 rows per bed, and a row spacing of 0.25m. The test greenhouse uses 1000kg / 667m special organic fertilizer for vegetables. 2 and compound fertilizer 50kg / 667m 2 The cultivation and management measures adopted the local routine management, and the strawberry grew well and evenly during the experiment.

[0158] Experimental treatment: 18% isoprene·thiopyrad suspension (Formulation Example 3) was set at 750, 1000, and 1500 times of dilution for three treatments, 5% thiopyrad suspension was set at 1000 times, 15% isoprene suspension was set at 1200 times and a total of 6 treatments for water control, each treatment was repeated 4 times, a total of 24 plots, each plot was 25m 2, , randomized block arrangement.

[0159] Application: The test was conducted on February 15, February 22, and March 1, 2023, for a total of 3 applications. A backpack manual sprayer was used with a spray hole diameter of 0.7 mm and a spray volume of 750 L / hm 2 , spray the strawberry plants evenly, without missing any spots or spraying too heavily, until the leaves are almost dripping.

[0160] Survey and record method: Sampling was conducted at 5 points on the diagonal of each plot, and 50 fruits were surveyed at each point to record the total number of fruits and the number of diseased fruits. Because the disease did not occur before the first application of pesticides, no disease base survey was conducted. The total number of fruits and the number of diseased fruits were surveyed 7 days after the first application of pesticides (February 22), 7 days after the second application of pesticides (March 1), and 7 days after the third application of pesticides (March 8), and the control effect was calculated. Duncan's new multiple range method (DMRT) was used for statistical analysis. The survey and grading standards for strawberry gray mold fruit are as follows:

[0161] Level 0: no disease;

[0162] Level 1: The lesion area accounts for less than 5% of the entire fruit surface;

[0163] Level 3: The lesion area accounts for 6% to 15% of the entire fruit area;

[0164] Level 5: The lesion area accounts for 16% to 25% of the entire fruit area;

[0165] Level 7: The lesion area accounts for 26% to 50% of the entire fruit area;

[0166] Level 9: The area of ​​lesions accounts for more than 50% of the entire fruit area.

[0167] The control effect of the pesticide was calculated according to the following formula, and the safety of the pesticide on strawberries was also investigated during the experiment.

[0168] Disease index = 100 × ∑ (number of diseased fruits at each level × representative value of each level) / (total number of fruits surveyed × highest representative value)

[0169] Control effect (%) = (blank control disease index - treatment area disease index) / blank control disease index × 100

[0170] The results are shown in Table 8:

[0171] Table 8 Statistics of field efficacy of each treatment agent against strawberry gray mold

[0172]

[0173] Note: Different lowercase letters in the same column represent significant differences among treatments (P<0.05).

[0174] As can be seen from Table 8, the three tested agents all have a certain control effect on strawberry gray mold. Among them, when the formulation example 3: 18% isoprene·thiophanate-methyl SC was diluted 750 times, the control effect of strawberry gray mold was more significant, with a control effect of 87.3%; at the same time, when the dilution multiple was 1000 times and 1500 times, the control effect was 83.58% and 81.62% respectively. Compared with 5% thiophanate-methyl SC and 15% isoprene SC, both have better control effects and a long lasting effect. In addition, after the third application of the drug, the strawberry leaves, young fruits, and mature fruits were observed to have no drug damage and no drug spots appeared, and the safety was good.

[0175] Example 4: Field efficacy test of the combination of isoprenaline and thiophanate-methyl against tomato gray mold

[0176] Selection of test site: The test was conducted in a greenhouse in Shen County, Liaocheng, Shandong Province. The previous crop was cucumber. The greenhouse area was 650m 2 The soil is sandy loam with medium to high fertility, medium organic matter content, flat terrain and good water conditions. The tomato variety for the test is "Jindi 855", which was planted on April 5, 2023, with a planting density of 4,000 plants / mu and equal row spacing. After planting, the management was as usual, and the tomatoes grew well and evenly.

[0177] Experimental treatment: 18% isoprene·thiopyrad suspension (Formulation Example 3) was set at 750, 1000, and 1500 times of dilution for three treatments, 5% thiopyrad suspension was set at 1000 times, 15% isoprene suspension was set at 1200 times and a total of 6 treatments for water control, each treatment was repeated 3 times, a total of 18 plots, each plot was 36m 2 , randomized block arrangement.

[0178] Application: During the trial, the pesticide was applied twice. The first application was on the morning of June 20, 2023, when tomato gray mold was in its early stages. The second application was on the morning of June 27, when the tomato fruiting period was approaching. A backpack manual sprayer was used to spray the plants evenly, with a spray rate of 1000 ppm every 667 m2. 2 The water consumption was 50 kg. No other pesticides were sprayed during the experiment. The growth of tomatoes in each plot was consistent before and after spraying, and other management measures were normal.

[0179] Investigation and recording method: Investigations were conducted before application, 7 days after the first application (June 27), 7 days after the second application (July 3), and 14 days after the second application (July 10). Since the fruit part was not diseased before application, the disease base before application only investigated the disease situation of the leaf part, and a total of 4 investigations were conducted. Each plot was randomly sampled by the 5-point sampling method, and 3 tomato plants were investigated at each point. The number of diseased leaves and fruits at each level was recorded, and the disease index and control effect of leaves and fruits were calculated. Finally, the experimental data were statistically analyzed using Duncan's new multiple range method (DMRT). In addition, it was observed whether there was any drug damage after application and the type and degree of drug damage were recorded.

[0180] The classification standards for tomato gray mold leaf survey are as follows:

[0181] Level 0: no lesions;

[0182] Level 1: The lesion area accounts for less than 5% of the entire leaf area;

[0183] Level 3: The lesion area accounts for 5% to 10% of the entire leaf area;

[0184] Level 5: The lesion area accounts for 10% to 20% of the entire leaf area;

[0185] Level 7: The lesion area accounts for 20% to 50% of the entire leaf area;

[0186] Level 9: The lesion area accounts for more than 50% of the total leaf area.

[0187] The investigation and grading standards for tomato gray mold fruit are as follows:

[0188] Level 0: no lesions;

[0189] Level 1: The remaining petals are diseased or the stigma is diseased;

[0190] Level 3: Sepal rot or column hair disease spreads to the navel;

[0191] Level 5: There are infiltration spots on the navel of the fruit without mold layer;

[0192] Grade 7: There is a mold layer at the navel but it does not spread to other parts;

[0193] Level 9: The mold layer extends to other parts of the fruit.

[0194] The control effect of the pesticide was calculated according to the following formula, and the safety of the pesticide to tomatoes was also investigated during the experiment.

[0195] Disease index = 100 × ∑ (number of diseased leaves or fruits at each level × representative value at each level) / (total number of fruits surveyed × highest representative value)

[0196] Fruit control effect (%) = (blank control disease index - treatment area disease index) / blank control disease index × 100

[0197]

[0198] Where: CK0——disease index of blank control area before application;

[0199] CK1——disease index of blank control area after application of pesticide;

[0200] PT0——disease index before application of pesticide in the treatment area;

[0201] PT1——Disease index after application of pesticide in the treatment area.

[0202] The results are shown in Tables 9 and 10:

[0203] Table 9 Statistics of field efficacy of each treatment agent in controlling tomato gray mold on leaves

[0204]

[0205] Table 10 Statistics on the field efficacy of each treatment agent in controlling tomato gray mold on the fruit part

[0206]

[0207] Note: Different lowercase letters in the same column represent significant differences among treatments (P<0.05).

[0208] As can be seen from Tables 9 and 10, the three tested agents all have certain control effects on tomato gray mold. Among them, when the formulation example 3: 18% isoprene·thiophanate-methyl SC was diluted 750 times, the control effect of tomato gray mold was more significant. After the second application of the drug for 14 days, the control effect on the leaf part was 84.31%, and the control effect on the fruit part was 86.67%; at the same time, when the dilution multiple was 1000 times and 1500 times, the control effect on leaves and fruits was 81.91%, 77.38% and 83.85%, 76.59%, respectively. Compared with 5% thiophanate-methyl SC and 15% isoprene SC, both had good control effects and significant differences, and the lasting effect was long. In addition, by comparing with CK, no drug damage or drug spots were observed on tomato leaves, young fruits, and mature fruits after the second application of the drug, and the plants grew healthily, indicating that the drug has good safety.

[0209] The present invention has shown through indoor toxicity tests and field efficacy tests that when the ratio of isoprene to thiamethoxam is 15:1 to 1:15, it has good control effects on lettuce sclerotinia, peanut brown spot, strawberry gray mold and tomato gray mold, and when the ratio is 5:1, the control effect on the above three diseases is the best and the effect is significant and the lasting effect is long. Secondly, after the application of the pesticide, there is no adverse effect on the growth of lettuce, peanut, strawberry and tomato.

[0210] The above-mentioned ideal embodiments of the present invention are used as examples. Through the description of the above-mentioned examples, a person skilled in the art can make various changes and modifications without departing from the technical concept of the present invention. The technical scope of the present invention is not limited to the contents in the specification, but must be determined according to the scope of the claims.

Claims

1. A fungicidal composition containing isoprenaline and thiophanate-methyl, characterized in that: The active ingredients in the bactericidal composition are isoprenaline and thiophanate-methyl, and the weight ratio of the two is 15:1 to 1:

15.

2. The bactericidal composition according to claim 1, characterized in that The weight ratio of isoprofenamide to thiophanate-methyl is 10:1 to 1:

10.

3. The bactericidal composition according to claim 1 or 2, characterized in that The total weight percentage of isoprenaline and thiophanate-methyl in the sterilization composition is 5% to 90%, and the rest are agricultural adjuvants allowed to be added and used in agricultural production.

4. The bactericidal composition according to claim 3, characterized in that The total weight percentage of the isoprenaline and thiophanate-methyl in the sterilization composition is 10% to 60%.

5. The bactericidal composition according to any one of claims 1 to 4, characterized in that The bactericidal composition can be prepared into any one of aqueous emulsion, microemulsion, suspension, water dispersible granule and wettable powder.

6. The bactericidal composition according to claim 5, characterized in that The bactericidal composition is preferably prepared as a suspension.

7. A method for controlling agricultural diseases using the fungicidal composition according to any one of claims 1 to 6, characterized in that: The bactericidal composition is brought into contact with crops or agricultural pathogens to effectively inhibit the growth of the pathogens until they die.

8. The method according to claim 7, characterized in that The crops are preferably lettuce, peanut, strawberry and tomato; the agricultural pathogens are preferably lettuce sclerotinia, peanut brown spot pathogen, strawberry gray mold and tomato gray mold.

9. Use of the fungicidal composition according to any one of claims 1 to 6 for preventing and controlling agricultural diseases.

10. The use according to claim 9, characterized in that The agricultural diseases are preferably lettuce sclerotinia rot, peanut brown spot, strawberry gray mold and tomato gray mold.

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