Application of cuminic acid in preparing nematicides and nematicide preparations

Abstract

The present invention discloses the use of cuminic acid for preparing nematicides and nematicide preparations. Research has found that cuminic acid and related nematicide preparations can be used not only to control the southern root-knot nematode Meloidogyne incognita (Kofold & White) Chitwood on crops such as tomatoes, pumpkins, and cucumbers, but also to control the potato stem nematode Ditylenchus destructor on crops such as potatoes and sweet potatoes. The preparation contains 0.1% to 1% cuminic acid, with the remainder being adjuvants. The nematicide of the present invention is not only effective in killing nematodes but also safe for the environment, humans, livestock, natural enemies of pests, and other beneficial organisms. The preparation process is simple, the cost is low, and it is suitable for widespread application.

Description

Technical Field

[0001] The invention belongs to the field of plants and their applications, and particularly relates to the application of cuminic acid in preparing a nematicide and the nematicide. Background Art

[0002] The southern root-knot nematode, Meloidogyne incognita (Kofold & White) Chitwood, belongs to the order Meloidogyne, family Heterodera, genus Meloidogyne. It is a major parasitic nematode with a wide host range, damaging hundreds of plant species. In mild cases, the root-knot nematode does not cause noticeable changes in the aboveground parts of the plant, making it difficult to detect. However, in severe cases, leaves turn yellow and wilt, the plant becomes stunted, and fruit yield and quality decline, sometimes even leading to plant death. Due to its strong adaptability, reproductive capacity, hidden nature, and resistance to pesticides, the control of root-knot nematodes is very difficult.

[0003] The potato nematode, Ditylenchus destructor Thorne, belongs to the order Ceratophyllum, family Graminaceae, genus Ditylenchus. It is an important parasitic nematode that attacks potatoes and sweet potatoes. However, in my country, the sweet potato is the primary target, and it occurs in both northern and southern potato regions. The potato nematode only infects the host's underground tissues. Infected sweet potatoes rot, dry out, shrink, crack, and darken internally. This is often accompanied by secondary infections with fungi, bacteria, and mites. Summary of the Invention

[0004] The present invention finds that cuminic acid and related nematicide preparations can be used not only for controlling the southern root-knot nematode Meloidogyne incognita (Kofold & White) Chitwood on crops such as tomatoes, pumpkins and cucumbers, but also for controlling the potato stem nematode Ditylenchus destructor on crops such as potatoes and sweet potatoes.

[0005] Based on this, the present invention provides the use of cuminic acid for preparing a nematicide preparation, wherein the nematodes are Ditylenchus potatoeus and / or Meloidogyne incognita.

[0006] The present invention also provides a corresponding nematocide preparation, which contains cuminic acid, and the mass percentage of cuminic acid is 0.1% to 1%. The nematocide preparation is in the form of a soluble liquid, a microemulsion or an emulsifiable concentrate preparation.

[0007] An optional solution is that the nematicide preparation is in the form of a microemulsion, and the microemulsion is prepared from the following raw materials in percentage by mass:

[0008] Cuminic acid 0.1% to 1%, organic solvent 0% to 30%, emulsifier 5% to 20%, antifreeze 5% to 10%, the balance being water, and the sum of the mass percentages of each raw material is 100%, excluding the endpoint value 0;

[0009] The organic solvent is selected from one or a mixture of two or more of ethyl acetate, cyclohexanone, methyl oleate, ethanol, methanol, acetone, ethylbenzene, N,N-dimethylformamide and xylene;

[0010] The emulsifier is a nonionic surfactant with a hydrophilic-lipophilic balance value of 12-18; for example: polyoxyethylene styrene acid ether, alkyl polyglycoside A, OP-10, concentrated milk 1601, polyoxyethylene (16EO) lanolin ether, calcium dodecylbenzenesulfonate, sodium dodecylbenzenesulfonate, ethylene glycol phenyl ether, fatty alcohol polyoxyethylene ether, etc.

[0011] The antifreeze agent is selected from one or a mixture of two or more of ethylene glycol, propylene glycol, glycerol and glycerin.

[0012] An optional solution is that the nematicide preparation is in the form of a soluble liquid, which is prepared from the following raw materials in percentage by mass:

[0013] Cuminic acid 0.1% to 1%, organic solvent 0% to 20%, surfactant 5% to 25%, antifreeze 3% to 10%, the balance being water, and the sum of the mass percentages of the raw materials is 100%, excluding the endpoint value 0;

[0014] The organic solvent is selected from one or a mixture of two or more of ethyl acetate, cyclohexanone, methyl oleate, ethanol, methanol, acetone, ethylbenzene, N,N-dimethylformamide and xylene;

[0015] The surfactant is selected from at least one of alkylbenzene sulfonates, alkylphenol polyoxyethylene ethers, phenethylphenol polyoxyethylene ethers, phenethylphenol polyoxyethylene ethers, polyoxypropylene ethers, adducts of castor oil and ethylene oxide, and arylalkylphenol polyoxyethylene ether formaldehyde condensates;

[0016] The antifreeze agent is selected from one or a mixture of two or more of ethylene glycol, propylene glycol, glycerol and glycerin.

[0017] An optional solution is that the nematicide preparation is in the form of an emulsifiable concentrate, which is prepared from the following raw materials in percentage by mass:

[0018] Cuminic acid 0.1% to 1%, organic solvent: 30% to 85%, emulsifier 10% to 30%, antifreeze 5% to 10%, the total mass percentage of the raw materials is 100%;

[0019] The organic solvent is selected from one or a mixture of two or more of ethyl acetate, cyclohexanone, methyl oleate, ethanol, methanol, acetone, ethylbenzene, N,N-dimethylformamide and xylene;

[0020] The emulsifier is a nonionic surfactant with a hydrophilic-lipophilic balance value of 12-18; for example: polyoxyethylene styrene acid ether, alkyl polyglycoside A, OP-10, concentrated milk 1601, polyoxyethylene (16EO) lanolin ether, calcium dodecylbenzenesulfonate, sodium dodecylbenzenesulfonate, ethylene glycol phenyl ether, fatty alcohol polyoxyethylene ether, etc.

[0021] The antifreeze agent is selected from one or a mixture of two or more of ethylene glycol, propylene glycol, glycerol and glycerin. DETAILED DESCRIPTION

[0022] Unless otherwise specified, the scientific and technical terms used herein are understood according to the knowledge of ordinary technicians in the relevant fields.

[0023] Cuminic acid (p-isopropylbenzoic acid, cuminic acid) is a small molecule organic acid isolated from cumin seeds. The present invention has found that cuminic acid has a good control effect on southern root-knot nematodes and potato stem nematodes. A certain proportion of adjuvants are further added to form the dosage form, and the dosage form can be an environmentally friendly preparation microemulsion, soluble liquid or emulsifiable concentrate. The inventor's experiments have shown that 0.1% cuminic acid microemulsion, 0.5% cuminic acid soluble liquid and 1.0% cuminic acid emulsifiable concentrate have a good control effect on southern root-knot nematodes and potato stem nematodes. In order to better understand the essence of the invention, the technical content of the invention is described in detail with examples below, but the invention is not limited to these examples. The substances and raw materials used in the following examples are all commercially available products.

[0024] Example 1: Indoor bioassay of cumic acid against nematodes:

[0025] The nematode activity of cuminic acid preparations was determined using the drug immersion method, with healthy, uniformly sized nematodes selected as test nematodes. The specific method is as follows:

[0026] The experiment was carried out in a 24-well cell culture plate. First, the same volume (V1) of the nematode suspension (ultrapure water, concentration: 1 nematode / μl) was injected into the sample well. Then, according to the test concentration [calculated according to formula (3)], sterile water (V2) was added as needed. Finally, the same volume of the drug mother solution (V3, specifically 50μl of the drug mother solution) with a series of concentrations was added. The mixture was evenly mixed and placed in a 25℃ incubator. Three replicates were performed for each treatment, with 50 to 100 nematodes per replicate. 50μl of methanol was used as a control.

[0027] 24h and 48h after treatment, the death and survival of nematodes in each treatment were checked, and the nematode mortality rate and the nematode adjusted mortality rate were calculated according to formula (1) and formula (2), respectively.

[0028] Method for judging whether nematodes are dead or alive: observe under a microscope. Those that are swimming and in an "S" shape are recorded as alive; for nematodes in a "straight line" or "half-moon" shape, use eyelashes to touch the nematode body repeatedly. If the nematode changes shape obviously after being touched, it is recorded as alive, otherwise it is recorded as dead.

[0029]

[0030] V = volume of nematode suspension V1 + volume of supplemented sterile water V2 + volume of drug stock solution V3; V = 400 μL; V1 = 100 μL; N is set in advance in the experiment, and in this embodiment, N = 400 mg / L; to ensure consistency in the number of treated nematodes, N0 is known, and in this embodiment, N0 = 400 mg / L, so V is fixed, and V2 and V3 of different treatments can generally be derived based on this.

[0031] The toxicity of cuminic acid to nematodes was determined by the drug solution immersion method, and the toxicity regression equation curve was drawn. The results are shown in Tables 1, 2 and 3.

[0032] The horizontal axis of the toxicity curve is the logarithm of the concentration, and the vertical axis is the probability value of the mortality rate. The data in each column shown in Table 3 were obtained by processing with DPS data processing software.

[0033] Table 1 Toxic effect of cuminic acid on Ditylenchus potatoe

[0034]

[0035] Table 2 Toxic effect of cumic acid on southern root-knot nematode

[0036]

[0037] Table 3 Toxicity curve of cuminic acid's nematode killing activity

[0038]

[0039] From the above table, we can see that cuminic acid has strong toxicity against Ditylenchus potatoeum and Meloidogyne incognita. When Ditylenchus potatoeum is treated with 400 mg / L, the corrected mortality rate after 72 hours is 91.95%; when Meloidogyne incognita is treated with 400 mg / L, the corrected mortality rate after 72 hours is 92.45%. 50 The LC values ​​of incognita root-knot nematode at 48h and 72h were 167.5211 and 78.0028 mg / L respectively. 50They are 168.7735 and 94.0782 mg / L respectively.

[0040] Example 2: Preparation of 0.1% cuminic acid microemulsion

[0041] 0.1 kg of cuminic acid was weighed and dissolved in 30 kg of methanol. 15 kg of calcium dodecylbenzenesulfonate, 5 kg of polyoxyethylene styrene ether, and 5 kg of ethylene glycol were then added and mixed under high-speed stirring. The mixture was heated to approximately 40°C. Deionized water was then added dropwise to the mixture until 100 kg was reached, with the addition rate controlled to maintain the temperature at approximately 40°C. After the addition of water was complete, the mixture was heated to 50°C and stirred for 1 hour to produce 100 kg of a 0.1% cuminic acid microemulsion. The stability and appearance of the preparation met the requirements for commercial preparations.

[0042] Example 3: Preparation of 0.5% cuminic acid soluble solution

[0043] Weigh 0.5 kg of cuminic acid and dissolve it in 20 kg of methanol. Add 15 kg of sodium dodecylbenzenesulfonate and 4 kg of propylene glycol while stirring. Make up to 100 kg with deionized water. Mix thoroughly and stir at 800-1000 rpm for 10-30 minutes to produce 100 kg of a 0.5% cuminic acid soluble liquid. The stability and appearance of the preparation meet the requirements of commercial preparations.

[0044] Example 4: Preparation of 1% cuminic acid emulsifiable concentrate

[0045] Weigh 1 kg of cuminic acid, 25 kg of ethanol, and 25 kg of ethyl acetate, then add 20 kg of sodium dodecylbenzenesulfonate, 5 kg of alkyl polyglycoside A, 10 kg of ethylene glycol, and make up to 100 kg with No. 150 solvent oil. After mixing thoroughly, stir at 800-1000 rpm for 10-30 minutes to produce 100 kg of cuminic acid emulsifiable concentrate. The stability and appearance of the preparation meet the requirements of commercial preparations.

[0046] Example 5: Example 2-4 Field test results of cuminic acid preparation against southern root-knot nematode

[0047] 1. Test reagents

[0048] Cuminic acid microemulsion, the cuminic acid microemulsion in Example 1;

[0049] Cuminic acid soluble liquid, Example 2 Cuminic acid soluble liquid;

[0050] Cuminic acid emulsifiable concentrate, the cuminic acid emulsifiable concentrate in Example 3;

[0051] 41.7% fluopyram suspension concentrate (Bayer CropScience (China) Co., Ltd. Beijing Branch);

[0052] 2. Test crops

[0053] Tomato, variety "Jinpin Maofen 802"

[0054] 3. Targets of prevention and control

[0055] Southern root-knot nematode (Meloidogyne incognita);

[0056] 4. Selection of experimental plots and experimental treatment

[0057] 4.1 Selection of experimental plots

[0058] This experiment was conducted in a greenhouse in Dali Modern Agricultural Park, Weinan City, Shaanxi Province. A representative tomato plot with disease symptoms was selected as the test site. The previous crop of tomatoes was planted, and the southern root-knot nematode occurred evenly, single and serious, while other diseases and pests were mild. The soil was sandy loam, with a light texture and high nutrient content, and normal fertilizer and water management was carried out.

[0059] 4.2 Experimental treatment and plot design

[0060] The test agents were 100× (“100×” means 100-fold dilution, the same below), 300×, and 500× cuminic acid microemulsion; 100×, 300×, and 500× cuminic acid soluble concentrate; 300× cuminic acid emulsifiable concentrate; 1500× 41.7% fluopyram suspension concentrate; and a blank control (water). The experimental plots were arranged in randomized blocks, with each plot measuring 25 m2. 2 , repeat 3 times.

[0061] 4.3 Application method

[0062] Before treatment, each plot was irrigated to ensure soil moisture reached 60%-70% of its saturated water holding capacity. Before application, the soil was tilled to a depth of 20-30 cm. The pesticide was sprayed evenly onto the soil surface at a rate of 40 liters / mu and tilled again to thoroughly mix the pesticide with the soil before transplanting the tomatoes. After application of the emulsifiable concentrate, the plots were quickly covered with plastic film and compacted with soil to prevent air leakage. The greenhouse doors were closed for 10 days, then opened and the film was removed for five days to ensure that no toxic gases remained in the soil before transplanting the tomatoes. A 1500× suspension concentrate of 41.7% fluopyram and a blank control were used, respectively, under the same conditions and with normal water and fertilizer management. During the experimental period, greenhouse temperatures were relatively low, reaching a maximum of 30°C and a minimum of 20°C. Samples were collected and investigated two months later.

[0063] 4.4 Survey Methods

[0064] Random sampling was used, with 5 points in each plot and 5 plants in each point. The number of root knots on each tomato plant was checked according to the following standards and the classification was recorded. The classification method is shown in Table 4 below:

[0065] Table 4 Root knot grading standards

[0066]

[0067] The control effect is calculated according to the following formula:

[0068] Root knot index (%) = {[∑(number of diseased plants × corresponding level value)] / total number of plants × 9} × 100; corresponding level values ​​are 0 / 1 / 3 / 5 / 7 or 9;

[0069] Control effect (%) = [(root knot index of the clear water control area - root knot index of the chemical treatment area) / root knot index of the control area] × 100;

[0070] 5. Experimental results and statistics:

[0071] The control effects of 0.1% cuminic acid microemulsion, 0.5% cuminic acid soluble solution and 1% cuminic acid emulsifiable concentrate on southern root-knot nematode were determined. The results are shown in Table 5.

[0072] Table 5 Control effects of three cuminic acid preparations on southern root-knot nematodes

[0073]

[0074]

[0075] The data in Table 5 are the averages of three replicates; in the "Effective" column, the same letter after the value in the same column means that the difference is not significant at the P = 0.05 level (LSD method). (Same as in the following tables)

[0076] As shown in Table 5, all three cuminic acid preparations showed good control efficacy against southern root-knot nematodes. The 0.5% cuminic acid soluble solution diluted 100-fold showed the best control efficacy, while the other treatments all achieved efficacy rates exceeding 60%. Therefore, the above three cuminic acid preparations are recommended.

[0077] Example 6: Field test results of Example 2-4 cuminic acid preparations against potato stem nematodes

[0078] 1. Test reagents

[0079] Cuminic acid microemulsion, cuminic acid microemulsion in Example 1

[0080] Cuminic acid soluble liquid, Example 2 Cuminic acid soluble liquid

[0081] Cuminic acid emulsifiable concentrate, cuminic acid emulsifiable concentrate in Example 3

[0082] 1.8% Abamectin EC (Jiangsu Suling Pesticide Chemical Co., Ltd.)

[0083] 2. Test crops

[0084] Potatoes, variety "Purple Flower White"

[0085] 3. Targets of prevention and control

[0086] Potato stem nematode (Ditylenchus destructor)

[0087] 4. Selection of experimental plots and experimental treatment

[0088] 4.1 Selection of experimental plots

[0089] This experiment was conducted in the greenhouse of Dali Modern Agricultural Park in Weinan City, Shaanxi Province. A representative potato plot with disease incidence was selected as the test site. The previous crop of potatoes was planted, and the potato stem nematode occurred evenly, single and serious, while other diseases and pests were mild. The soil was sandy loam, with a light texture and high nutrient content, and normal fertilizer and water management was carried out.

[0090] 4.2 Experimental treatment and plot design

[0091] The test agents were 100×, 300×, and 500× cuminic acid microemulsion; 100×, 300×, and 500× cuminic acid soluble concentrate; 300× cuminic acid emulsifiable concentrate; 1000× 1.8% avermectin emulsifiable concentrate and a blank control (water). The experimental plots were arranged in randomized blocks, with each plot having an area of ​​25 m 2 , repeat 3 times.

[0092] 4.3 Application method

[0093] Before treatment, each plot was irrigated to ensure soil moisture reached 60%-70% of its saturated water holding capacity. Before application, the soil was tilled to a depth of 20-30 cm. The pesticide was evenly sprayed onto the soil surface at a rate of 40 liters / mu and tilled again to thoroughly mix the pesticide with the soil before planting. Following application of the emulsifiable concentrate, the plots were quickly covered with plastic film and compacted with soil to prevent air leakage. The greenhouse doors were closed for 10 days, then opened and the film was removed for five days to ensure that no toxic gases remained in the soil before planting. A 1.8% avermectin emulsifiable concentrate (1000x) and a water treatment served as control and blank controls, respectively, with the same water and fertilizer management practices. Temperatures within the greenhouse remained low during the experiment, reaching a maximum of 30°C and a minimum of 20°C. Potato samples were collected after harvest.

[0094] 4.4 Survey Methods

[0095] After 45 days, sampling was carried out using the "Z"-shaped sampling method, with 5 sampling points in each area. The sampling depths were 0-10 cm, 10-20 cm, and 20-30 cm, and the soil drill diameter was 2.5 cm. 200 mg of soil was taken each time, and the shallow plate method was used to separate the nematodes. The nematodes were killed with warm water at 60°C and fixed with formaldehyde, and the nematode population reduction rate was calculated.

[0096] During the potato harvest period, samples were taken at five points in each plot of the experimental area. Ten potato tubers were taken at each point, and each tuber was cross-sectioned and graded according to the disease severity of the cross-section:

[0097] Level 0: no symptoms (no hollows or cracks);

[0098] Grade 1: The affected area occupies less than 25% of the cross-section;

[0099] Grade 2: The affected area accounts for 25% to 50% of the cross-sectional area;

[0100] Grade 3: The affected area accounts for 50% to 75% of the cross-section;

[0101] Level 4: The affected area occupies more than 75% of the cross-section.

[0102] Record the number of tubers at each level and calculate the disease index and control effect. The calculation formula is as follows:

[0103] Disease index = [∑(disease grade value × number of tubers of corresponding grade)] / (total number of tubers surveyed × highest disease grade value) × 100;

[0104] Control effect = (disease index in control area - number of diseased areas in treatment area) / disease index in control area

[0105] ×100%;

[0106] 5. Experimental results and statistics

[0107] The control effects of 0.1% cuminic acid microemulsion, 0.5% cuminic acid soluble liquid and 1% cuminic acid emulsifiable concentrate on potato stem nematode were determined. The results are shown in Table 6.

[0108] Table 6 Control effects of three cumin extract nematicides on Ditylenchus potatoein

[0109]

[0110] The data in Table 6 are the averages of three replicates; in the "Effect" column, the same letter after the value in the same column means that the difference is not significant at the P = 0.05 level (LSD method)

[0111] As shown in Table 6, all three cuminic acid preparations demonstrated significant efficacy against Ditylenchus potatoevarii, with no significant differences in efficacy compared to the control agent. The efficacy of a 100-fold dilution of 0.1% cuminic acid microemulsion, a 100-fold dilution of 0.5% cuminic acid soluble solution, and a 300-fold dilution of 1% cuminic acid emulsifiable concentrate (EC) all exceeded 80%, significantly exceeding the efficacy of the control agent, 1.8% avermectin EC. Furthermore, the efficacy of these preparations at different dilutions was above 60%, exceeding or comparable to that of the control agent, 1.8% avermectin EC. Therefore, the above three cuminic acid preparations are recommended for the control of Ditylenchus potatoevarii.

[0112] The preferred embodiments of the present disclosure are described in detail above. However, the present disclosure is not limited to the specific details of the above embodiments. Within the technical concept of the present disclosure, various simple modifications can be made to the technical solutions of the present disclosure, and these simple modifications all fall within the scope of protection of the present disclosure.

[0113] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present disclosure will not further describe various possible combinations.

[0114] In addition, the various embodiments of the present disclosure may be arbitrarily combined, and as long as they do not violate the concept of the present disclosure, they should also be regarded as the contents disclosed by the present disclosure.

Claims

1. Use of cuminic acid in preparing a nematode preparation, wherein the nematodes are Ditylenchus potatoeus and / or Meloidogyne incognita.

2. The use according to claim 1, characterized in that The nematocidal preparation contains cuminic acid, and the mass percentage of cuminic acid is 0.1% to 1%. The nematocidal preparation is in the form of a soluble liquid, a microemulsion or an emulsifiable concentrate preparation.

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