Application of bactericidal agricultural composition containing WML-01 and methoxyl acrylate medicament in prevention and treatment of plant diseases

Abstract

The invention discloses an application of a bactericidal agricultural composition containing WML-01 and a methoxy acrylate medicament (QoIs) in prevention and treatment of rice bakanae disease, wheat stem rot, wheat scab, plant root rot, rice blast, plant anthracnose and sclerotinia rot of colza. In the bactericidal agricultural composition containing the WML-01 and the methoxyl acrylate medicament, the mass ratio of the WML-01 to the methoxyl acrylate medicament is (10: 1) to (1: 10). The bactericidal agricultural composition can be used for preventing and treating rice bakanae disease, wheat stem rot, wheat scab, plant root rot, rice blast, plant anthracnose and sclerotinia rot of colza, and can effectively solve the problems of few medicaments for preventing and treating the seven diseases, unstable prevention effect, high prevention and treatment cost and drug resistance in the current production; wide market application prospects are realized.

Description

TECHNICAL FIELD

[0001] The application relates to an application of a bactericidal agricultural composition containing WML-01 and a methoxy acrylate in plant disease control BACKGROUND

[0002] Rice seedling blight, also known as the excessive growth disease, is an important disease in rice production. The main pathogen is Fusarium fujikuroi, and the seed and field disease residues are the main initial infection sources of the rice seedling blight. The diseased rice seeds are often not germinated or cannot be sprouted after sowing. Under normal circumstances, the diseased seedlings at the seedling stage are thinner and taller than the healthy seedlings, the leaf sheath is long and thin, the leaf color is light yellow, and the root system is poorly developed. Some diseased seedlings die before transplanting. After the diseased seedlings are planted, the mycelium can expand to the whole plant under suitable conditions, and stimulate the stem and leaf to grow excessively. During the flowering period, the pathogen spreads to the flowers and invades the lemma and endosperm, causing the grain to be abnormal or deformed. If the pathogen invades late, the grain may not show symptoms, but the mycelium has already invaded the interior, making the seed carry the pathogen, which greatly affects the planting of the next crop of rice. Chemical control is still the main means for the prevention and control of the rice seedling blight. At present, the bactericides registered for the prevention and control of the rice seedling blight on the market include carbendazim, prochloraz, difenoconazole and kresoxim-methyl. However, due to the long-term, large-scale and frequent use of chemical agents, the pathogen gradually develops resistance to different bactericides, resulting in a significant decrease in the prevention and control effect of the agents. Increasing the amount of the agents will cause problems such as pesticide residues, seed safety and environmental pollution.

[0003] Wheat stem base rot, commonly known as "crown rot" or "dry land foot disease", is a global wheat stem base disease caused by multiple fusariums such as fusarium pseudograminearum and fusarium graminearum, and the fusarium pseudograminearum is the main dominant pathogen. The early stage of wheat stem base rot can cause the wheat seedling root or embryo sheath part to be brown and rotten, and the serious wheat seedling root or embryo sheath part can even lead to the death of the wheat seedling. Subsequently, the diseased part gradually expands to the first and second stem nodes of the wheat, causing the diseased part to become dark brown or rotten, the stem base leaf sheath to be dry, and the stem to be brown and necrotic. In the later stage, on the basis of the brown stem base, the water transport in the vascular bundle from the root to the ear is hindered, causing the wheat ear to appear "ear" white symptoms, and after the formation of "white ear", the wheat grain cannot continue to develop. The disease greatly affects the seed setting rate of wheat and seriously affects the yield of wheat. At present, only 8 pesticide products have been registered for the prevention and control of the disease, and the pesticide varieties are only difenoconazole, tebuconazole, kresoxim-methyl, prothioconazole, trifloxystrobin and nitomefen. There is an urgent need for new agents for the prevention and control of the disease.

[0004] Wheat scab, also known as "rot head wheat", is one of the world's wheat diseases, caused by a variety of Fusarium infection, among which F. graminearum is the dominant pathogen in many wheat producing areas. Scab causes seedling blight, stem rot and ear rot, and the diseased grains also contain toxins, which can cause poisoning after eating. The main fungicides for controlling wheat scab in production include benzimidazole fungicides, triazole fungicides and methoxy acrylate fungicides, etc. The long-term and large-scale use of chemical fungicides has led to the increasingly prominent problem of wheat scab resistance.

[0005] Root rot is a common disease in plant production, and the pathogen mainly damages the roots of plants, causing poor plant growth, seed rot, root rot, dwarfing, yellowing, and even whole plant death, resulting in serious yield loss. Soybean root rot is a major root disease in soybean production. The disease occurs in seedling stage, affecting the growth of seedlings and even causing seedling death, resulting in a decrease in the number of seedlings in the field. In the plant stage, the root damage affects the growth and number of root nodules, causing poor growth of the aboveground part, dwarfing, affecting pod number and grain weight, and thus leading to yield reduction. Currently, the fungicides registered for the control of soybean root rot on the market include difenoconazole, prochloraz, azoxystrobin, carbendazim, and thiram, etc.

[0006] Rice blast, also known as rice fever, fire blast, and head blast, is one of the important diseases of rice, widely distributed, and the pathogen is Pyricularia oryzae Cav. Rice blast mainly includes seedling blast, leaf blast, node blast, panicle neck blast, and grain blast, with node blast and panicle neck blast being the most harmful. The disease can cause 10% to 30% yield reduction, 40% to 50% yield reduction, or even complete yield loss. The main fungicides for controlling rice blast in production include tricyclazole, carbendazim, isoprothiolane, prochloraz, azoxystrobin, and validamycin, etc. The long-term and large-scale use of chemical fungicides has led to the increasingly prominent problem of rice blast resistance.

[0007] Anthracnose is a common disease of plants, mainly damaging leaves, fruits, and branches, causing concave lesions, and the surface of the lesions often has small black spots (conidia discs), which seriously affects the yield and quality of plant products. The pathogen causing anthracnose is mainly the fungus of the genus Colletotrichum spp., among which C. gloeosporioides is a representative species, which can cause anthracnose of citrus, mango, pomegranate, grape, strawberry, pepper, tomato, ginger, onion, rubber, corn, sugarcane, and many flowers. The main agents for controlling anthracnose at present include methoxy acrylate, triazole, and benzimidazole, and there are many pesticide products, but the types of pesticide varieties are concentrated, and the number of products containing difenoconazole and azoxystrobin accounts for more than 25% of the total number of products. The long-term and concentrated use of chemical agents has led to the increasingly prominent problem of anthracnose resistance.

[0008] Sclerotinia sclerotiorum, which can infect stems, leaves, flowers, and silique of rape, among which the stem is most seriously affected. After the onset of the disease, water-stained, light brown lesions appear on the stems, and the central part of the lesion turns white, with the edge being brown. The lesion is clearly demarcated from the healthy part. White, fluffy mycelium grows on the lesion when it is wet, eventually leading to stem rot, breakage, and death of the whole plant.

[0009] Strobilurin fungicides are widely used in the prevention and treatment of agricultural diseases. They are derived from mushroom extracts and exert their effects by inhibiting the cytochrome bc1 complex in the mitochondrial respiratory chain of pathogenic fungi. These fungicides have high fungicidal activity and good systemicity, and are effective against a variety of fungal diseases and oomycete diseases. Currently, the commonly used strobilurin fungicides include azoxystrobin, pyraclostrobin, trifloxystrobin, and enoxastrobin. With years of frequent use, many diseases have developed resistance to strobilurin fungicides, such as gray mold, early blight, downy mildew, and powdery mildew.

[0010] There are few fungicides for the prevention and treatment of rice bakanae disease, wheat foot rot, wheat scab, plant root rot, rice blast, plant anthracnose, and rape sclerotinia. With the long-term use of single-mechanism selective fungicides, pathogenic fungi have developed resistance to varying degrees. The development of new fungicides with novel mechanisms and their rational combination with strobilurin fungicides through scientific compounding modes can reduce the dosage of fungicides, improve their efficacy, delay the development of resistance, and extend the life cycle of the fungicides, which is of great significance for disease control and prolonging the life cycle of the fungicides. SUMMARY

[0011] The purpose of the present application is to provide a new use of a bactericidal agricultural composition containing WML-01 and a strobilurin fungicide in the prevention and treatment of plant diseases.

[0012] To achieve the purpose of the present application, the WML-01 provided by the present application is a butenolide compound containing a thiazolidine structure, which has the following structural formula:

[0013]

[0014] The compound has a broad fungicidal spectrum and good control effect on major crop fungal diseases (wheat scab, wheat foot rot, rice bakanae disease, rice blast, cotton fusarium wilt, grape anthracnose, rape sclerotinia, and soybean root rot). The compound is disclosed in patent publication CN 118724887 A, and the compound number is 3-1.

[0015] The inventors of the present application found that WML-01 has good in vitro bacteriostatic activity against various plant pathogenic fungi, and has no cross-resistance with the commonly used bactericides in current production, and it is speculated that it is a new type of bactericide with a completely new action target site and good application prospect.

[0016] The bactericidal agricultural composition of WML-01 and azoxystrobin provided by the present application has obvious synergistic and additive effects on the prevention and control of rice sheath blight, wheat foot rot, wheat scab, rice blast, plant root rot, sclerotinia rot and anthracnose, can effectively prevent and control diseases, reduce the amount of drug, meet the demand of "zero growth of pesticides", and different action mechanisms of the mixed agent are important measures for international fungicide resistance management, which is beneficial to delay the development of new drug resistance.

[0017] Specifically, WML-01 and azoxystrobin have a co-toxicity coefficient of 89.35-195.947 against rice sheath blight fungus in a compounding range of 10:1-1:10 (w / w), showing additive and synergistic effects. In particular, when the mass ratio of WML-01 and azoxystrobin is 10:1-1:1, the co-toxicity coefficient is greater than 120, and the synergistic effect is significant.

[0018] WML-01 and azoxystrobin are compounded in a mass ratio of 10:1-1:10 (w / w), and the co-toxicity coefficient against wheat foot rot fungus is determined by the co-toxicity coefficient method to be 92.78-173.13, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 and azoxystrobin is 10:1-1:3, the co-toxicity coefficient is greater than 120, and the synergistic effect is significant.

[0019] WML-01 and azoxystrobin are compounded in a mass ratio of 10:1-1:10 (w / w), and the co-toxicity coefficient against wheat foot rot fungus is determined by the co-toxicity coefficient method to be 99.72-165.14, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 and azoxystrobin is 10:1-1:1, the co-toxicity coefficient is greater than 120, and the synergistic effect is significant.

[0020] WML-01 and azoxystrobin are compounded in a mass ratio of 10:1-1:10 (w / w), and the co-toxicity coefficient against wheat foot rot fungus is determined by the co-toxicity coefficient method to be 99.72-165.14, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 and azoxystrobin is 10:1-1:1, the co-toxicity coefficient is greater than 120, and the synergistic effect is significant.

[0021] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against rice blast fungus were determined by the co-toxicity coefficient method, ranging from 88.04 to 190.91, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 2:1, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0022] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against Sclerotinia sclerotinia, the causal agent of rapeseed, were determined by the co-toxicity coefficient method, ranging from 106.25 to 240.53, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:2, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0023] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against *Anthracnose causal agent* of pepper were determined by the co-toxicity coefficient method, ranging from 83.17 to 144.55, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 3:1 to 2:1, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0024] Furthermore, the present invention provides a fungicide agricultural composition containing WML-01 and pyraclostrobin, wherein the mass ratio of the fungicide components WML-01 and pyraclostrobin in the agricultural composition is 10:5.

[0025] As will be readily apparent to those skilled in the art, in order to fully utilize the efficacy of pesticide active ingredients, they can be formulated into easily usable formulations according to usage requirements. Based on this, the present invention further provides a fungicidal agricultural composition containing WML-01 and azoxystrobin, wherein the agricultural composition with WML-01 and azoxystrobin as active ingredients is processed into an agriculturally acceptable formulation, wherein the formulation components include adjuvants or carriers that enhance the efficacy of WML-01 and azoxystrobin.

[0026] The fungicide agricultural composition can be formulated into various dosage forms such as seed coating agents, suspensions, water-in-oil emulsions, microemulsions, emulsifiable concentrates, wettable powders, water-dispersible granules, microcapsule suspensions, nano-formulations, and granules.

[0027] The fillers include, but are not limited to, bentonite, silica, magnesium aluminum silicate, etc., and the additives include, but are not limited to, xanthan gum, wetting agents, dispersants, defoamers, antifreeze agents, warning colors, etc.

[0028] The wetting agents include, but are not limited to, nonylphenol polyoxyethylene ether NP-10, alkylphenol polyoxyethylene ether OP-10, alkyl naphthalene sulfonate EFW, etc.; the dispersants include, but are not limited to, sodium salt of alkyl naphthalene sulfonate condensate D-425, sulfonate of alkyl naphthalene sulfonate condensate D-450, etc.; the defoamers include, but are not limited to, n-octanol, organosilicon SAG1522, etc.; the antifreeze agents include, but are not limited to, ethylene glycol, etc.; and the warning colors include, but are not limited to, basic rose essence, golden red 3110, and sun-resistant peach red 3228, etc.

[0029] The present invention relates to the use of a fungicide agricultural composition containing WML-01 and pyraclostrobin in the control of plant diseases. As described above, within a suitable dosage range, the fungicide agricultural composition exhibits excellent disease control effects. The application methods of the fungicide agricultural composition and its formulations are diverse, and the present invention does not impose any particular limitation. One or a combination of various application methods can be used, such as seed coating, seed soaking, spraying, root drenching, broadcasting, hole application, and furrow application. Depending on crop cultivation, formulation form, and target pests, seed coating, seed dressing, and seed soaking are preferred methods for this fungicide agricultural composition.

[0030] Compared with existing technologies, the present invention provides a fungicidal agricultural composition with WML-01 and methoxyacrylate agents as active ingredients. Within the mass ratio range of 10:1 to 1:10, the present invention exhibits synergistic and additive effects against rice bakanae disease, wheat stem base rot, wheat scab, plant root rot, rice blast, plant anthracnose, and rapeseed sclerotinia stem rot. The use of this fungicidal composition can effectively improve disease control, reduce pesticide usage, delay the development of pesticide resistance, and increase crop yield. In summary, this fungicidal composition is safe, effective, and economical, and has broad market application prospects. Detailed Implementation

[0031] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0032] Example 1: In vitro antibacterial activity of WML-01 and azoxystrobin against rice bakanae disease pathogens, wheat stem rot pathogens, wheat scab pathogens, rice blast pathogens, soybean root rot pathogens, rapeseed sclerotinia rot pathogens, and pepper anthracnose pathogens.

[0033] 1. Test materials

[0034] 1.1 Test strains

[0035] The rice bakanae disease pathogen (F. f. f. f. f. f. f.) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0036] The soybean root rot pathogen (F. oxysporum) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0037] The fungus causing wheat stem rot (F. pseudoograminearum) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0038] The fungus causing wheat scab (F. graminearum) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0039] Rice blast fungus (P. oryzae) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0040] The sclerotiorum causal agent of rapeseed (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0041] The anthracnose fungus of pepper (C. gloeosporioides) (provided by the College of Plant Protection, China Agricultural University) was identified using conventional methods.

[0042] 1.2 Test reagents

[0043] WML-01 (99% technical grade), provided by the College of Science, China Agricultural University. This compound and its preparation method are disclosed in patent publication number CN 118724887 A, compound number 3-1.

[0044] Its structural formula is:

[0045]

[0046] Azoxystrobin (98% technical grade), a product of Adama Anbang (Jiangsu) Co., Ltd.

[0047] The above-mentioned pesticide technical material was prepared into a concentration of 10 using dimethyl sulfoxide as a solvent. 4 The stock solution of mg / L was stored in a refrigerator at 4°C for later use.

[0048] 1.3 Test Culture Medium

[0049] Potato glucose medium (PDA): 200g potato, 18g glucose, 12g agar powder, 1000mL distilled water.

[0050] 2. Experimental Methods

[0051] The test was conducted according to the plate method in the Guidelines for Pesticide Bioassay in the Laboratory - Fungicides - Part 2: Test on Inhibiting the Mycelial Growth of Pathogenic Fungi, NY / T 1156.2 - 2006. The main operation process of the plate method is as follows: Five concentration gradients were set between 10% and 90% of the inhibition rate of the agent against the target bacteria. Then, a punch with a diameter of 6 mm was used to punch out fungal discs on the same circumference near the colony edge of the activated target bacteria (ensuring that the ages of the test bacteria in the same replicate are the same), and the fungal discs were inoculated onto the centers of a series of culture medium plates containing the agent under aseptic conditions with an inoculation needle, with the mycelial side facing downwards, and a non - agent - added plate was set as a control, and it was cultured in the dark. Each treatment was replicated 3 times for determination. The colony diameters of each concentration treatment were recorded, and the mycelial growth inhibition rate was calculated. Then, the inhibition rate was converted into a probit value (Y), and the agent concentration was converted into a logarithm to the base 10 (X). The X - Y was used to make a regression line, and the virulence regression curve equation Y = a + bX of each agent against the target bacteria, as well as the correlation coefficient r and the effective median inhibitory concentration EC, were obtained respectively. 50 .

[0052] The combined toxicity of the compound preparation is expressed by the co - toxicity coefficient method of Sun Yunpei:

[0053] The theoretical toxicity index TTI of the compound preparation = ∑(toxicity index of a certain drug × percentage of the active ingredient of this drug in the mixture).

[0054]

[0055] When CTC ≥ 120, it is a synergistic effect; when CTC ≤ 80, it is an antagonistic effect; when 80 < CTC < 120, it is an additive effect.

[0056] Test results

[0057] Table 1. In vitro antibacterial activity of the mixture of WML - 01 and azoxystrobin against Fusarium moniliforme of rice

[0058]

[0059]

[0060] Table 2. In vitro antibacterial activity of the mixture of WML - 01 and azoxystrobin against Rhizoctonia cerealis of wheat

[0061]

[0062] Table 3. In vitro antibacterial activity of the mixture of WML - 01 and azoxystrobin against Gibberella zeae of wheat

[0063]

[0064] Table 4. In vitro antibacterial activity of the mixture of WML - 01 and azoxystrobin against Fusarium solani of soybean

[0065]

[0066]

[0067] Table 5. In vitro antibacterial activity of WML-01 and azoxystrobin against rice blast fungus.

[0068]

[0069] Table 6. In vitro antibacterial activity of WML-01 and azoxystrobin combined against Sclerotinia sclerotinia, the causal agent of rapeseed rot.

[0070]

[0071]

[0072] Table 7. In vitro antibacterial activity of WML-01 and azoxystrobin combined against *Anthracnose causal agent*.

[0073]

[0074] The experimental results (see Tables 1-7) show that the co-toxicity coefficients of WML-01 and azoxystrobin in the compound range of 10:1 to 1:10 (w / w) against rice bakanae disease pathogens ranged from 89.35 to 195.947, exhibiting additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:1, the co-toxicity coefficients were all greater than 120, demonstrating a significant synergistic effect.

[0075] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against wheat stem rot pathogens were determined by the co-toxicity coefficient method, ranging from 92.78 to 173.13, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:3, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0076] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against Fusarium graminearum, the causal agent of wheat blight, were determined by the co-toxicity coefficient method, ranging from 99.72 to 165.14, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:1, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0077] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against soybean root rot pathogens were determined by the co-toxicity coefficient method, ranging from 97.22 to 172.68, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:3, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0078] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against rice blast fungus were determined by the co-toxicity coefficient method, ranging from 88.04 to 190.91, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 2:1, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0079] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against Sclerotinia sclerotinia, the causal agent of rapeseed, were determined by the co-toxicity coefficient method, ranging from 106.25 to 240.53, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 10:1 to 1:2, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0080] WML-01 and azoxystrobin were combined at a mass ratio of 10:1 to 1:10 (w / w). The co-toxicity coefficients against *Anthracnose causal agent* of pepper were determined by the co-toxicity coefficient method, ranging from 83.17 to 144.55, showing additive and synergistic effects. In particular, when the mass ratio of WML-01 to azoxystrobin was 3:1 to 2:1, the co-toxicity coefficients were all greater than 120, indicating a significant synergistic effect.

[0081] Example 2: Field control efficacy of WML-01 and azoxystrobin combined with wheat scab.

[0082] I. Materials and Methods

[0083] 1.1 Test Materials

[0084] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, product of Adama Anbang (Jiangsu) Co., Ltd.

[0085] 1.2 Overview of the test site

[0086] The experiment was conducted in Fengtai County, Anhui Province, in a clay loam soil with moderate fertility. The wheat variety was Huaiyumai No. 1, and the seed rate was 25 kg per mu.

[0087] 1.3 Experimental Design

[0088] Table 8. Experimental scheme for seed treatment against wheat scab.

[0089]

[0090] 1.4 Survey Methods and Calculations

[0091] Five random sampling points were taken from each treatment area, with 100 ears of panicle at each point. The disease index and control effect were calculated based on the percentage of panicle dead area to the total panicle area.

[0092] Grading standards for Fusarium head blight:

[0093] Grade 0, disease-free throughout the ear of grain;

[0094] Grade 1: The area of ​​dead ears accounts for less than 1 / 4 of the total ear area;

[0095] Grade 3, with withered ears covering 1 / 4 to 1 / 2 of the total ear area;

[0096] Grade 5, withered ears covering 1 / 2 to 3 / 4 of the total ear area;

[0097] Grade 7, with withered ears covering more than 3 / 4 of the total ear area;

[0098] Methods for calculating drug efficacy:

[0099] Disease index (%) = ∑(Number of diseased plants (ears) at each level × Representative value at each level) / (Total number of plants (ears) surveyed × Highest representative value) × 100

[0100] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0101] II. Test Results

[0102] Table 9. Efficacy of pesticide treatments in controlling wheat scab.

[0103] Note: * The result is the average of four replicates; the significance level for the analysis of variance is p = 0.05.

[0104] The experimental results (Table 9) show that after two applications, the high-dose 200ml treatment (30g active ingredient) of 15% WML-01·pyraclostrobin suspension showed an efficacy of 89.51% against wheat scab, which was significantly better than the single-agent treatment (10g active ingredient) and the single-agent treatment (20g active ingredient) of WML-01; the low-dose 100ml treatment (15g active ingredient) showed an efficacy of 86.23% against wheat scab, which was significantly better than the single-agent treatment of WML-01 and the single-agent treatment of pyraclostrobin.

[0105] When WML-01 and pyraclostrobin are combined, they show excellent control efficacy against wheat scab and significantly reduce the amount of pesticide used in the field.

[0106] Example 3: Field control efficacy of WML-01 and azoxystrobin combined with soybean root rot

[0107] 1. Materials and Methods

[0108] 1.1 Test Materials

[0109] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, produced by Adama Anbang (Jiangsu) Co., Ltd.

[0110] 1.2 Overview of the test site

[0111] The experimental site was located in the Beian Modern Agricultural Demonstration Park. The soil type was black soil, with an organic matter content of 5.6% and a pH of 6.6. Sowing was carried out on May 21, 2024, using a 65cm ridge-three cultivation pattern, maintaining 320,000 plants per hectare. 48% soybean-specific fertilizer was applied at a rate of 320 kg per hectare (N:P:K = 15:23:10). Emergence occurred on June 1. Soil moisture was good. The previous crop was soybean. Irrigation relied on natural rainfall.

[0112] 1.3 Experimental Design

[0113] Table 10 Experimental protocols for seed treatments against plant root rot

[0114]

[0115] 1.4 Survey Methods and Calculations

[0116] Soybean root rot survey

[0117] Surveys were conducted twice, at 30 days and 60 days after soybean emergence. At least 30 soybean plants were randomly selected from each plot for investigation, and the total number of plants and the number of plants with diseases at each level were recorded. The incidence of root rot was investigated, and the effectiveness of control measures was calculated.

[0118] Disease classification: (area of ​​lesions on the roots)

[0119] Grade 0: Main root and fibrous roots are intact, with no disease spots and many root nodules;

[0120] Grade 1: Scattered lesions on the main root, but not in clusters; lesions on the fibrous roots for several days.

[0121] Grade 3: The main root shows numerous lesions, but the lesions cover less than 1 / 4 of the root area; the fibrous roots are slightly affected.

[0122] Grade 5: The lesions on the main root cover an area of ​​1 / 4 to 1 / 2 of the root area. There are many fibrous roots affected, but they do not form large patches.

[0123] Grade 7: The area of ​​lesions on the main root is between 1 / 2 and 1 / 4 of the root area, and the lesions on the fibrous roots are in patches, with some fibrous roots falling off;

[0124] Level 9: The entire root system is surrounded by disease spots, the root is rotten, and there are almost no fibrous roots.

[0125] Methods for calculating drug efficacy:

[0126] Disease index (%) = ∑(Number of diseased plants (ears) at each level × Representative value at each level) / (Total number of plants (ears) surveyed × Highest representative value) × 100

[0127] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0128] II. Test Results

[0129] Table 11. Control efficacy of pesticide treatments against soybean root rot.

[0130]

[0131] Note: * The average of 4 repetitions; ** The significance level for the analysis of variance was p = 0.05.

[0132] The experimental results (Table 11) showed that, 30 days after emergence, the high-dose seed treatments of 200ml (30g active ingredient) and 100ml (15g active ingredient) of 15% WML-01·pyraclostrobin suspension had control efficacy of 82.29% and 75.72% against soybean root rot, respectively, which were significantly better than those of WML-01 (20g active ingredient) and pyraclostrobin (25g active ingredient).

[0133] A survey conducted 60 days after sowing showed that seed treatments with 100ml and 200ml of 15% WML-01·pyraclostrobin suspension exhibited excellent control efficacy against soybean root rot, with efficacy rates of 81.46% and 85.82%, respectively, significantly superior to single-agent treatments with WML-01 and pyraclostrobin.

[0134] When WML-01 and pyraclostrobin are combined, they show excellent control efficacy against soybean root rot, and the amount of pesticide used in the field is significantly reduced.

[0135] Example 4: Field control efficacy of WML-01 and azoxystrobin combined with wheat stem rot

[0136] 1. Materials and Methods

[0137] 1.1 Test Materials

[0138] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% Witconol NP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% Witconol NP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, produced by Adama Anbang (Jiangsu) Co., Ltd.

[0139] 1.2 Overview of the test site

[0140] The experiment was conducted in Feixiang District, Handan City, Hebei Province. The soil was clay loam with moderate fertility. The wheat variety was Woyu No. 3, and the seed rate was 20 kg per mu.

[0141] 1.3 Experimental Design

[0142] Table 12 Experimental protocols for seed treatments against wheat stem rot

[0143]

[0144] 1.4 Survey Methods and Calculations

[0145] Stem base rot survey

[0146] a) Survey on the efficacy of stem base rot control during the grain-filling stage

[0147] Five random sampling points were taken from each treatment area, with 100 plants at each point. The number of diseased plants and the disease index were investigated, and the disease index and control effect were calculated.

[0148] Grading standards for stem base rot:

[0149] Grade 0, the plant is not diseased;

[0150] Grade 1, the stem in the ground is noticeably brown;

[0151] Level 3, the first intersegment turns brown;

[0152] Level 5, second interphalangeal joint turns brown;

[0153] Level 7, browning symptoms appeared in the third internode, but no white ears were observed.

[0154] Level 9: White ears or plant death due to disease.

[0155] Methods for calculating drug efficacy:

[0156] Disease index (%) = ∑(Number of diseased plants (ears) at each level × Representative value at each level) / (Total number of plants (ears) surveyed × Highest representative value) × 100

[0157] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0158] b) Survey of white ear rate during grain-filling period

[0159] Five random sampling points were taken from each treatment area, with 100 plants at each point. The total number of ears and the number of white ears were investigated, and the control effect was calculated.

[0160] White ear percentage (%) = Number of white ears / Total number of plants surveyed × 100

[0161] Control efficacy (%) = [White ear rate in control area - White ear rate in treated area] / White ear rate in control area × 100

[0162] II. Test Results

[0163] Table 13. Control efficacy of pesticide treatments against wheat stem rot.

[0164]

[0165]

[0166] Note: * The average of 4 repetitions; ** The significance level for the analysis of variance was p = 0.05.

[0167] The experimental results (Table 13) show that, during the grain-filling stage, the high-dose (200ml) seed dressing treatment of 15% WML-01·pyraclostrobin suspension (30g active ingredient) achieved a control efficacy of 92.54% against wheat stem base rot, while the low-dose (100ml) treatment (15g active ingredient) achieved a control efficacy of 88.93%. Both were significantly superior to the single-agent seed dressing treatment of pyraclostrobin (25g active ingredient) and the single-agent treatment of WML-01 (20g active ingredient).

[0168] The results of the white ear rate survey showed that the high-dose (200ml) treatment of 15% WML-01·pyraclostrobin suspension achieved a white ear control efficacy of 96.55%; the low-dose (100ml) treatment achieved a white ear control efficacy of 86.04%. This was superior to WML-01 single agent treatment and significantly superior to pyraclostrobin single agent treatment.

[0169] When WML-01 and pyraclostrobin are combined, they show excellent control efficacy against wheat stem rot, and the amount of pesticide used in the field is significantly reduced.

[0170] Example 5: Field control efficacy of WML-01 and azoxystrobin combined with rice bakanae disease

[0171] 1. Materials and Methods

[0172] 1.1 Test Materials

[0173] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, produced by Adama Anbang (Jiangsu) Co., Ltd.

[0174] 1.2 Overview of the test site

[0175] The experiment was conducted in Zhaoyuan Village, Yaojiang Town, Zhuji City, Zhejiang Province. The soil was clay loam with moderate fertility. The rice variety was Zhongzao 39, and the seed rate was 5 kg per mu. Seedlings were raised and transplanted.

[0176] 1.3 Experimental Design

[0177] Table 14 Experimental protocols for seed treatment against rice bakanae disease

[0178]

[0179] 1.4 Survey Methods and Calculations

[0180] Bakanae disease: Five random sampling points were taken from each plot in the seedbed, with each sampling point measuring 10cm*10cm. The sampling was repeated three times. The disease incidence rate was recorded, and the control efficacy was calculated.

[0181] Disease incidence rate (%) = (Number of diseased plants / Total number of plants surveyed) × 100%

[0182] Control efficacy (%) = (Disease rate in the blank control area - Disease rate in the treatment area) / Disease rate in the blank control area × 100%

[0183] 2. Experimental Results

[0184] Table 15. Control efficacy of pesticide treatments against rice bakanae disease.

[0185]

[0186] Note: * The average of 4 repetitions; ** The significance level for the analysis of variance was p = 0.05.

[0187] The experimental results (Table 15) showed that during the seedling stage, the high-dose seed treatments of 200ml (30g active ingredient) and 100ml (15g active ingredient) of 15% WML-01·pyraclostrobin suspension achieved 100% control efficacy against rice bakanae disease, which was significantly better than the single-agent treatment of pyraclostrobin (12.5g) and the single-agent treatment of WML-01 (20g active ingredient).

[0188] The combination of WML-01 and pyraclostrobin showed excellent control efficacy against bakanae disease, and significantly reduced the amount of pesticide used in the field.

[0189] Example 6: Field control efficacy of WML-01 and azoxystrobin combined with rice blast.

[0190] I. Materials and Methods

[0191] 1.1 Test Materials

[0192] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, product of Adama Anbang (Jiangsu) Co., Ltd.

[0193] 1.2 Overview of the test site

[0194] The experiment was conducted in Chadi Township, Shanghang County, Longyan City, Fujian Province, using the rice variety Changliangyou 8.

[0195] 1.3 Experimental Design

[0196] Table 16 Experimental protocols for pesticide treatments against rice blast

[0197]

[0198] 1.4 Survey Methods and Calculations

[0199] A five-point sampling method was used, with at least 20 panicles sampled at each point. Each plot consisted of 100 panicles, and the total number of panicles, the number of diseased panicles, and the disease severity were recorded. The disease index and control effectiveness were calculated.

[0200] The grading standards for rice neck blast are as follows:

[0201] Level 0: No disease;

[0202] Level 1: Less than 5% loss per ear (individual branches affected);

[0203] Level 3: 6%–20% loss per ear (about one-third of the branches and stalks are affected);

[0204] Grade 5: 21%–50% loss per ear (disease on the neck or main axis, grains half-empty);

[0205] Level 7: 51%–70% loss per ear (neck disease, most ears are empty);

[0206] Grade 9: 71% to 100% loss per ear (caused by disease at the neck of the ear, resulting in white ears).

[0207] Methods for calculating drug efficacy:

[0208] Disease index (%) = ∑(Number of diseased plants (ears) at each level × Representative value at each level) / (Total number of plants (ears) surveyed × Highest representative value) × 100

[0209] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0210] II. Test Results

[0211] Table 17. Efficacy of pesticide treatments in controlling rice blast.

[0212]

[0213] Note: * The result is the average of four replicates; the significance level for the analysis of variance is p = 0.05.

[0214] The experimental results (Table 17) show that after two applications, the high-dose (200ml) treatment (30g active ingredient) of 15% WML-01·pyraclostrobin suspension showed an efficacy of 89.99% against rice blast, significantly better than the single-agent treatment (12.5g active ingredient) and the single-agent treatment (20g active ingredient) of WML-01. The low-dose (100ml) treatment (15g active ingredient) showed an efficacy of 80.15%, slightly better than the single-agent treatments of pyraclostrobin and WML-01.

[0215] When WML-01 is combined with pyraclostrobin, it shows excellent control efficacy against rice blast, and the amount of pesticide used in the field is significantly reduced.

[0216] Example 7: Field control efficacy of WML-01 and azoxystrobin combined with rapeseed sclerotinia stem rot

[0217] I. Materials and Methods

[0218] 1.1 Test Materials

[0219] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, product of Adama Anbang (Jiangsu) Co., Ltd.

[0220] 1.2 Overview of the test site

[0221] The experiment was conducted in Dafasi Town, Wuxue City, Huanggang City, Hubei Province, in a loamy soil.

[0222] The rapeseed variety Huayouza 62 was sown at a rate of 400g / mu.

[0223] 1.3 Experimental Design

[0224] Table 18 Experimental protocols for chemical treatments against Sclerotinia stem rot in rapeseed.

[0225]

[0226] 1.4 Survey Methods and Calculations

[0227] A five-point sampling method was used, with at least 40 plants sampled at each point, and 200 plants surveyed per plot. The number of diseased plants and the disease severity were recorded for each plot. The disease index and control effect were calculated.

[0228] The grading criteria are as follows:

[0229] Level 0: No disease;

[0230] Grade 1: The affected area accounts for less than 5% of the surface area of ​​the main stem;

[0231] Level 3: The diseased area accounts for more than 5% but less than 15% of the surface area of ​​the main stem;

[0232] Level 5: The diseased area accounts for more than 15% but less than 30% of the main stem surface area;

[0233] Level 7: The diseased area accounts for more than 30% but less than 50% of the surface area of ​​the main stem;

[0234] Level 9: The diseased area accounts for more than 50% of the surface area of ​​the main stem.

[0235] Methods for calculating drug efficacy:

[0236] Disease index (%) = ∑(Number of diseased plants at each level × Representative value at each level) / (Total number of plants surveyed × Highest representative value) × 100

[0237] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0238] II. Test Results

[0239] Table 19. Efficacy of pesticide treatments in controlling sclerotinia stem rot in rapeseed.

[0240]

[0241] Note: * The result is the average of four replicates; the significance level for the analysis of variance is p = 0.05.

[0242] The experimental results (Table 19) show that after two applications, the high-dose (100ml) treatment of 15% WML-01·pyraclostrobin suspension (15g active ingredient) achieved a control efficacy of 89.96% against Sclerotinia sclerotinia in rapeseed, significantly better than the single-agent treatment of pyraclostrobin (7.5g active ingredient) and the single-agent treatment of WML-01 (20g active ingredient). The low-dose (50ml) treatment (7.5g active ingredient) achieved a control efficacy of 82.76% against Sclerotinia sclerotinia in rapeseed, better than the single-agent treatment of pyraclostrobin and significantly better than the single-agent treatment of WML-01.

[0243] When WML-01 and pyraclostrobin are combined, they show excellent control efficacy against sclerotinia stem rot in rapeseed, and the amount of pesticide used in the field is significantly reduced.

[0244] Example 8: Field control efficacy of WML-01 and pyraclostrobin combined with pepper anthracnose.

[0245] I. Materials and Methods

[0246] 1.1 Test Materials

[0247] 15% WML-01 azoxystrobin suspension concentrate, self-made, comprising 10% WML-01, 5% azoxystrobin, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, prepared by sand milling process; 10% WML-01 suspension concentrate, self-made, comprising 10% WML-01, 3% MorwetD-425, 1% WitconolNP-100, 0.3% xanthan gum, and water balance, % being mass percentage, prepared by sand milling process; 25% azoxystrobin suspension concentrate, product of Adama Anbang (Jiangsu) Co., Ltd.

[0248] 1.2 Overview of the test site

[0249] The experiment was conducted in Daxin Town, Fugou County, Zhoukou City, Henan Province, in loam soil, using the chili pepper variety Mibao 19.

[0250] 1.3 Experimental Design

[0251] Table 20 Experimental protocols for chemical treatments against anthracnose in peppers

[0252]

[0253] 1.4 Survey Methods and Calculations

[0254] Five samples were taken from each area, and 20 fruits were investigated at each point. The fruits were graded based on the percentage of diseased area to the total fruit area.

[0255] Grading method:

[0256] Grade 0: No lesions;

[0257] Grade 1: The area of ​​lesions is less than 5% of the total fruit area;

[0258] Grade 3: The area of ​​lesions accounts for more than 5% but less than 10% of the total fruit area;

[0259] Grade 5: The area of ​​lesions accounts for more than 10% but less than 25% of the total fruit area;

[0260] Grade 7: The area of ​​lesions accounts for more than or equal to 25% and less than 50% of the total fruit area;

[0261] Level 9: The area of ​​lesions accounts for more than 50% of the total fruit area, or the fruit falls off.

[0262] Methods for calculating drug efficacy:

[0263] Disease index (%) = ∑(Number of diseased fruits at each level × Representative value at each level) / (Total number of fruits surveyed × Highest representative value) × 100

[0264] Control efficacy (%) = (Disease index after treatment in control area - Disease index after treatment in treatment area) / Disease index after treatment in control area × 100

[0265] II. Test Results

[0266] Table 21. Efficacy of chemical treatments in controlling anthracnose in peppers.

[0267]

[0268] Note: * The result is the average of four replicates; the significance level for the analysis of variance is p = 0.05.

[0269] The experimental results (Table 21) show that after two applications, the 200ml treatment (30g active ingredient) of 15% WML-01·pyraclostrobin suspension showed an efficacy of 88.04% against anthracnose in peppers, significantly better than the single-agent treatment (7.5g active ingredient) and the single-agent treatment (20g active ingredient) of WML-01. The 100ml treatment (7.5g active ingredient) showed an efficacy of 79.26% against anthracnose, also better than the single-agent treatments of pyraclostrobin and WML-01.

[0270] When WML-01 and pyraclostrobin are combined, they show excellent control efficacy against anthracnose in peppers, and the amount of pesticide used in the field is significantly reduced.

[0271] Although the present invention has been described in detail above with general descriptions and specific embodiments, 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 fall within the scope of protection claimed by the present invention.

Claims

1. A bactericidal composition comprising WML-01 and a methoxyacrylate agent, wherein the structural formula of WML-01 is shown in Formula I:

2. The bactericidal composition according to claim 1, characterized in that, The methoxyacrylate agent is selected from one of azoxystrobin, pyraclostrobin, oxadiazon, and pyrimethanil.

3. The bactericidal composition according to claim 1, characterized in that, The mass ratio of WML-01 to methoxyacrylate agents is 10:1 to 1:

10.

4. The bactericidal composition according to claim 1, characterized in that, The methoxyacrylate agent is azoxystrobin, and the mass ratio of WML-01 to azoxystrobin is 10:1 to 1:10, preferably 10:1 to 1:2, more preferably 10:1 to 1:1, particularly preferably 3:1 to 2:1, and most preferably 10:

5.

5. A fungicide formulation, wherein the active ingredient is the fungicide composition according to any one of claims 1-3, wherein the fungicide composition is processed into an agriculturally acceptable formulation, the formulation including seed coating agents, suspensions, water-in-oil emulsions, microemulsions, emulsifiable concentrates, microcapsule suspensions, nanoformulations, granules, wettable powders, or water-dispersible granules; wherein the fungicide formulation components include adjuvants or carriers that enhance the efficacy of WML-01 and methoxyacrylate agents.

6. The application of the bactericidal composition according to any one of claims 1-3 or the bactericidal agent according to claim 5 in the prevention and control of plant diseases.

7. The application according to claim 6, characterized in that, The plant diseases mentioned are one or more of the following: rice bakanae disease (Fusarium fujikuroi), wheat stem base rot (Fusarium pseudograminearum), wheat scab (Fusarium graminearum), plant root rot (Fusarium spp.), rice blast (Pyricularia oryzae), plant anthracnose (Coletotrichum spp.), and rapeseed sclerotinia sclerotiorum.

Images