Compound composition of carvacrol and hinokitiol, bactericide and application

Abstract

The invention provides a compound composition of carvacrol and hinokitiol, a bactericide and application. The mass ratio of the carvacrol to the hinokitiol is (1-8): (6-1). The carvacrol and the natural product hinokitiol are compounded, so that the tomato fusarium wilt can be prevented and treated, and powdery mildew, gray mold and the like of various crops can be prevented and treated. The binary compound agent has a remarkable synergistic effect on preventing and treating the diseases, the use amount of a single agent can be remarkably reduced, and the compound agent is expected to be developed into a new generation of green agricultural botanical fungicide.

Description

Technical Field

[0001] The present invention belongs to the field of agricultural fungicides, and specifically relates to a compound composition of carvacrol and hinokitiol, a fungicide and its application. The two compounds are binary compounded to prevent and treat various crop diseases such as tomato wilt, crop powdery mildew and gray mold, and belong to the invention of new use of fungicides and their application technology. Background Art

[0002] Crop diseases cause serious economic losses to the world's food production every year. The application of chemical fungicides is an important means of plant disease prevention and control at this stage. Due to the long-term single use of chemical fungicides, plant pathogens have become more and more serious, such as resistance to fungicides and excessive pesticide residues. Mixing or compounding pesticides can not only complement each other, expand the prevention and control spectrum, improve drug efficacy, reduce costs, reduce labor, and save costs, but more importantly, it can delay the occurrence and spread of pathogen resistance to drugs.

[0003] Carvacrol, also known as 2-methyl-5-isopropylphenol, is a volatile monoterpenoid phenol. Studies have shown that carvacrol is widely present in a variety of Lamiaceae and Verbenaceae plants, such as oregano, thyme, mint and winter campanula. Carvacrol has a simple structure and is easy to synthesize artificially. At present, carvacrol is widely used in food preservation, agricultural and forestry crop protection, antioxidant, anti-inflammatory and anti-tumor activities. According to the China Pesticide Information Network, carvacrol has been registered as a botanical fungicide for the prevention and control of tobacco virus disease, tomato gray mold and potato late blight and other crop diseases.

[0004] Hinokitiol is a natural tropolone compound extracted from the cypress tree. It is not only environmentally friendly, but also a good metal chelator. So far, researchers have discovered that hinokitiol has multiple activities, such as a wide range of bacterial and fungal inhibitors, a preservative for postharvest vegetables and fruits, an excellent wood preservative, a good heart protection drug, anti-cancer and anti-tumor capabilities, and an additive for cosmetics and shower gels. Our research group's previous studies have found that hinokitiol has a wide range of biological activities, especially for the prevention and control of soil-borne diseases. Because hinokitiol has a simple structure, is easy to synthesize, is environmentally friendly, and is safe for non-target organisms, it is a pure natural, safe, and efficient substance with great potential for development as a botanical fungicide. Summary of the invention

[0005] The present invention aims to provide a binary compound of carvacrol and the natural product hinokitiol, which can be used for preventing and treating tomato wilt, crop powdery mildew, gray mold and other crop diseases at the same time.

[0006] To achieve the above object, the technical solution adopted by the present invention includes:

[0007] A composite composition of carvacrol and hinokitiol, wherein the mass ratio of carvacrol to hinokitiol is (1-8): (6-1).

[0008] Optionally, carvacrol:hinokitiol=(1-3):(2-1).

[0009] Optionally, carvacrol:hinokitiol=4:1.

[0010] The use of any composite composition of carvacrol and hinokitiol described in the present invention for preparing a plant fungicide.

[0011] Optionally, by mass percentage, the bactericide contains 0.1% to 30% carvacrol and 0.1% to 30% hinokitiol.

[0012] Optionally, the plant fungicide is used to prevent and control tomato wilt, tomato gray mold, strawberry powdery mildew and / or strawberry gray mold.

[0013] A carvacrol and hinokitiol compound fungicide, wherein the active ingredient of the fungicide is any compound composition of carvacrol and hinokitiol described in the present invention;

[0014] Calculated by mass percentage, the bactericide contains 0.1% to 30% carvacrol and 0.1% to 30% hinokitiol.

[0015] Optionally, the fungicide is an aqueous emulsion, and the mass percentage is:

[0016] Carvacrol 0.1% to 30%, hinokitiol 0.1% to 30%, emulsifier 5% to 10%, dispersant 1% to 10%, antifreeze 0.5% to 5%, preservative 0.5% to 5%, solvent 5% to 15%, water to make up to 100%;

[0017] The dispersant is polyvinyl alcohol, gum arabic, xanthan gum or magnesium aluminum silicate;

[0018] The solvent is at least one of sec-butyl acetate, isoamyl citrate, ethyl acetate, cyclohexanone, furfural, 5-methylfurfural, ethanol and ethyl acetate;

[0019] The antifreeze agent is ethylene glycol, propylene glycol, glycerin or urea;

[0020] The preservative is sorbic acid, benzoic acid or benzaldehyde;

[0021] The emulsifier is at least one of alkyl polyoxyethylene ether, fatty acid polyoxyethylene ester, TX-10, 7781T, 4103, OP-10, Well201, Well205 and M30.

[0022] Optionally, the bactericide is a soluble liquid, and the mass percentage is:

[0023] Carvacrol 0.1% to 30%, hinokitiol 0.1% to 30%, emulsifier 5% to 25%, antifreeze 1% to 5%, and solvent to make up to 100%;

[0024] The emulsifier is at least one of phenylethylphenol polyoxyethylene ether, TX-10, 2201, Ningru 700, Nongru 500, Nongru 600, Nongru BY, Nongru EL and NP-30;

[0025] The solvent is at least one of methanol, ethanol, cyclohexanone, ethyl acetate, sec-butyl acetate, furfural and 5-methylfurfural;

[0026] The antifreeze agent is ethylene glycol, propylene glycol, glycerin or urea.

[0027] Compared with the prior art, the advantages of the present invention are:

[0028] (1) The present invention has found through a series of ratio studies that when carvacrol and hinokitiol are mixed in a mass ratio of 1:6 to 8:1, the synergistic coefficient is greater than 1.5, and both have obvious synergistic effects.

[0029] (2) The binary compound agent has significantly enhanced efficacy in controlling the above-mentioned diseases and can significantly reduce the amount of a single agent used. It is expected to be developed into a new generation of green agricultural plant-derived fungicide.

[0030] (3) Currently, there is only one pesticide registered on the China Pesticide Information Network for the prevention and treatment of tomato wilt. There is a shortage of pesticides for the prevention and treatment of tomato wilt. This compound preparation has a good prevention and treatment effect on tomato wilt and has great potential to be registered as a pesticide for the prevention and treatment of tomato wilt. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] The compound fungicide of carvacrol and hinokitiol of the present invention is a preparation compounded with carvacrol and hinokitiol in a mass ratio of 1:6 to 8:1, which can not only prevent and treat tomato wilt, but also can be used to prevent and treat powdery mildew and gray mold of various crops. The binary compound agent has significant synergy in preventing and treating the above diseases, and can significantly reduce the usage of a single agent. The compound preparation is expected to be developed into a new generation of green agricultural plant-derived fungicides.

[0033] For example, the dosage forms of fungicides include aqueous emulsions and soluble liquids;

[0034] The components and weight percentages of the aqueous emulsion are: 0.1% to 30% of carvacrol, 0.1% to 30% of hinokitiol, 5% to 10% of emulsifier, 1% to 10% of dispersant, 0.5% to 5% of antifreeze, 0.5% to 5% of preservative, 5% to 15% of solvent, and deionized water is used to make up to 100%; polyvinyl alcohol, gum arabic, xanthan gum or magnesium aluminum silicate is used as a dispersant; one or more of sec-butyl acetate, isoamyl citrate, ethyl acetate, cyclohexanone, furfural, 5-methylfurfural, ethanol and ethyl acetate is used as a solvent; ethylene glycol, propylene glycol, glycerol or urea is used as an antifreeze; sorbic acid, benzoic acid or benzaldehyde is used as a preservative; one or more of alkyl polyoxyethylene ether, fatty acid polyoxyethylene ester, TX-10, 7781T, 4103, OP-10, Well201, Well205 and M30 is used as an emulsifier.

[0035] The components and weight percentages of the soluble liquid are: 0.1% to 30% of carvacrol, 0.1% to 30% of hinokitiol, 5% to 25% of emulsifier, 1% to 5% of antifreeze, and the solvent is made up to 100%; one or more of TX-10, phenethylphenol polyoxyethylene ether, 2201, Ningru 700, Nongru 500, Nongru 600, Nongru BY, Nongru EL, NP-30, etc. are emulsifiers; one or more of methanol, ethanol, cyclohexanone, ethyl acetate, sec-butyl acetate, furfural, 5-methylfurfural, etc. are solvents; one or more of ethylene glycol, propylene glycol, glycerin, urea, etc. are antifreeze.

[0036] Example 1: Toxicity of carvacrol and hinokitiol to tomato wilt pathogen

[0037] The mycelium growth rate method was used to determine the toxicity of carvacrol and hinokitiol to pathogens. The original drugs of carvacrol and hinokitiol were dissolved in methanol to a series of concentrations. The mother solution was pipetted with a pipette and added to the PDA medium to prepare drug-containing medium of different concentrations. A bacterial cake with a diameter of 0.5 cm of pathogens was picked and placed in the above drug-containing medium. The inoculated culture dish was placed in a 25°C incubator for culture, and the colony diameters were measured and the inhibition rate was calculated using the cross method. The effective inhibition concentration (EC 50 ).

[0038] Table 1 Toxicity test of different ratios of carvacrol and hinokitiol against tomato wilt pathogen

[0039]

[0040] As shown in Table 2, the EC values ​​of carvacrol and hinokitiol against tomato wilt pathogens are 50The concentrations of carvacrol and hinokitiol were 35.1868 μg / mL and 19.1762 μg / mL, respectively, indicating that carvacrol and hinokitiol had good antibacterial activity against tomato Fusarium wilt.

[0041] Example 2: Synergistic effect of different ratios of carvacrol and hinokitiol

[0042] The toxicity of carvacrol and hinokitiol to pathogens at different ratios was determined by referring to the mycelium growth rate method. The original drugs of carvacrol and hinokitiol were dissolved in methanol to the same concentration of mother liquor, and the mixture was made according to different volume ratios. A series of concentrations of drug-containing culture medium were prepared according to Example 1. The bacterial cake was transferred to a PDA plate containing a series of concentrations of compounded drugs and cultured at 25°C for 2 constant temperature cultures. When the colony diameter of the blank control group exceeded 2 / 3 of the diameter of the entire culture dish, all colony diameters were measured. Each treatment was repeated 3 times, and the mycelium growth inhibition rate was calculated. The effective inhibition concentration (EC 50 ), the synergistic coefficient (SR) of the compound agent was calculated using the following formula according to the Wadley method. When SR is greater than 1.5, it is a synergistic effect; SR between 0.5 and 1.5 is an additive effect; SR less than 0.5 is an antagonistic effect (a and b are the proportions of carvacrol and hinokitiol in the compound, EC 50 (A) and EC 50 (B) is an effective medium concentration of carvacrol and hinokitiol, EC 50 (Exp) is the theoretical inhibitory concentration, EC 50 (Obs) is the actual measured inhibition median concentration).

[0043]

[0044] Table 2 Toxicity test of different ratios of carvacrol and hinokitiol against tomato wilt pathogen

[0045]

[0046]

[0047] The data in Table 2 show that different ratios of carvacrol and hinokitiol have different toxicity to tomato wilt pathogens, and the synergistic coefficient values ​​are between 0.8855 and 2.3659. The synergistic coefficients of the above ratios are all greater than 0.5, indicating that the combined use of carvacrol and hinokitiol in the above ratios exhibits an additive effect, especially when the ratio of carvacrol to hinokitiol is 1:6 to 8:1, the synergistic coefficients are all greater than 1.5, indicating that in the above ratios, carvacrol and hinokitiol have obvious synergistic effects on tomato wilt pathogens.

[0048] Example 3: Production of 10% carvacrol and 20% hinokitiol composition aqueous emulsion

[0049] Accurately weigh 102 kg of 98% carvacrol and 204 kg of 98% hinokitiol. Mix the two active ingredients in 2M 3 Mix thoroughly in the mixing tank. Accurately weigh 10 kg of sorbic acid, put it into the mixing tank, and mix thoroughly again. Then accurately add 80 kg of emulsifier fatty acid polyoxyethylene ester into the mixing tank, then add 10 kg of gum arabic, 10 kg of propylene glycol, 100 kg of ethyl acetate, add water to make up to 1000 kg, and mix thoroughly in the mixing tank. Continue stirring for 1 hour. Prepare 1 ton of water emulsion product of carvacrol and hinokitiol composition.

[0050] Sampling analysis showed that the content of carvacrol was 10% and the content of hinokitiol was 20%. The product was packaged into 500 mL double barrier glass bottles, sealed, and labeled to obtain a water emulsion product of the composition with a content of 30%.

[0051] Example 4: Production of a soluble liquid composition of 10% carvacrol and 20% hinokitiol

[0052] Accurately weigh 102 kg of 98% carvacrol and 204 kg of 98% hinokitiol. Mix the two active ingredients in 2M 3 Mix thoroughly in a blending tank. Slowly add the premixed active ingredient mixture to 400 kg of ethanol solvent, stirring continuously until the active ingredient is completely dissolved to form a uniform transparent solution. Then slowly add 244 kg of TX-10 emulsifier to the solution in which the active ingredient is completely dissolved, add 50 kg of antifreeze glycerin during the addition process, and then continue to stir to make it evenly dispersed. After the emulsifier is added, gradually increase the stirring speed to high speed and perform an emulsification operation for 30 minutes. A stable emulsified system is formed by high-speed stirring, and the active ingredient is evenly dispersed in the solvent to obtain a soluble liquid with good stability and fluidity. A soluble liquid product of 1 ton of carvacrol and hinokitiol composition is prepared.

[0053] Sampling analysis showed that the content of carvacrol was 10% and the content of hinokitiol was 20%. The product was packaged into 500 mL double barrier glass bottles, sealed, and labeled to obtain a soluble liquid product with a composition content of 30%.

[0054] Example 5: Effect of 30% aqueous emulsion and soluble liquid of carvacrol and hinokitiol on the prevention of tomato wilt and gray mold

[0055] Greenhouse pot experiments were conducted to determine the efficacy of 30% aqueous emulsion and soluble liquid on tomato wilt and gray mold. Three concentrations were set for the test agents. Bacillus amyloliquefaciens water-dispersible granules were used as the positive control agent for the prevention and control of tomato wilt, and 5% carvacrol aqueous solution was used as the positive control agent for the prevention and control of tomato gray mold. Each treatment was repeated in 5 pots. The tomato wilt inoculation and disease index survey were conducted with reference to the literature report of Luo Man et al.

[0056] Table 3 The efficacy of carvacrol and hinokitiol compound against tomato wilt

[0057]

[0058] Note: According to Fisher's least significant difference test, different letters after the values ​​in the same column indicate that there are significant differences between the values ​​(P=0.05).

[0059] As shown in Table 3, the greenhouse pot experiment proved that the water emulsion and soluble liquid of carvacrol and hinokitiol compound had good control effects on tomato wilt. When the dosage of the active ingredient of the compound preparation was 100 μg / mL, there was no significant difference in the control effect of tomato wilt with the control effect of the control agent; when the dosage was 200 μg / mL, the control effect of the compound preparation on tomato wilt was significantly better than that of the control agent. At the same dosage, the control effect of the soluble liquid of the compound preparation was slightly higher than that of the water emulsion.

[0060] The design of the tomato gray mold prevention test is as follows. After the seedling stage and before the flowering stage of tomatoes, a concentration of 5x 10 6 cfu / mL of tomato gray mold spore suspension was evenly sprayed on tomato leaves with a backpack sprayer. After 48 hours, different agents were sprayed with a backpack sprayer. Six tomato plants were selected for each treatment, and repeated 3 times. The disease condition of the plants was investigated 5 to 7 days after inoculation, and 10 leaves were collected from top to bottom for each plant to investigate the disease. The disease classification standard refers to GB / T17980.28-2000.

[0061] Table 4 The efficacy of carvacrol and hinokitiol compound against tomato gray mold

[0062]

[0063] Note: According to Fisher's least significant difference test, different letters after the values ​​in the same column indicate that there is a significant difference between the values ​​(P = 0.05). Control effect (%) = (control lesion diameter - treatment lesion diameter) / (control lesion diameter) * 100.

[0064] As shown in Table 4, the compound preparation of carvacrol and hinokitiol also has a good control effect on tomato gray mold. When the dosage of the compound preparation is 50μg / mL, its control effect is more than 50%. The control effect of the compound preparation of 100μg / mL is better than the control effect of the same dose of the control agent. The control effect of the compound preparation of 200μg / mL is significantly higher than that of the control agent. At the same dose, the control effect of the soluble liquid of the compound preparation is also slightly higher than the control effect of the water emulsion at the same dose.

[0065] Example 6: Effect of 30% aqueous emulsion and soluble liquid of carvacrol and hinokitiol on the prevention of strawberry powdery mildew

[0066] The efficacy test of compound 30% water emulsion and soluble liquid on strawberry powdery mildew was carried out in a greenhouse. The test agents were set at three concentrations of 100, 200, and 400 μg / mL. 30% fenthion wettable powder was used as the positive control agent for the prevention and control of strawberry powdery mildew. Four replicates were set for each concentration treatment, and 50 young fruits were randomly surveyed in each replicate, for a total of 200 fruits. The incidence of young fruits was investigated 7 days after application and the control effect was calculated.

[0067] Table 5 The efficacy of carvacrol and hinokitiol compound against strawberry powdery mildew

[0068]

[0069] Note: According to Fisher's least significant difference test, different letters after the values ​​in the same column indicate that there are significant differences between the values ​​(P=0.05).

[0070] As shown in Table 5, the compound preparation of carvacrol and hinokitiol also has a good control effect on strawberry powdery mildew. When the dosage of the compound preparation is 100 and 200 μg / mL, the control effect of different dosage forms is more than 55% and 75% respectively; when the compound preparation is 400 μg / mL, the control effect is more than 85%, and there is no significant difference with the control effect of the control agent. At the same dosage, the control effect of the soluble liquid of the compound preparation is equivalent to that of the water emulsion, and there is no obvious pattern.

[0071] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A composite composition of carvacrol and hinokitiol, characterized in that: By mass ratio, carvacrol:hinokitiol = (1-8): (6-1).

2. The composite composition of carvacrol and hinokitiol according to claim 1, characterized in that: Carvacrol:hinokitiol = (1-3): (2-1).

3. The composite composition of carvacrol and hinokitiol according to claim 1, characterized in that: Carvacrol:hinokitiol = 4:

1.

4. Use of the composite composition of carvacrol and hinokitiol according to any one of claims 1 to 3 for preparing a plant fungicide.

5. The use according to claim 4, characterized in that: Calculated by mass percentage, the bactericide contains 0.1% to 30% carvacrol and 0.1% to 30% hinokitiol.

6. The use according to claim 4, characterized in that: The plant fungicide is used for preventing and controlling tomato wilt, tomato gray mold, strawberry powdery mildew and / or strawberry gray mold.

7. A compound fungicide of carvacrol and hinokitiol, characterized in that: The active ingredient of the fungicide is a composite composition of carvacrol and hinokitiol as described in any one of claims 1 to 3; Calculated by mass percentage, the bactericide contains 0.1% to 30% carvacrol and 0.1% to 30% hinokitiol.

8. The composite fungicide of carvacrol and hinokitiol according to claim 7, characterized in that: The fungicide is a water emulsion, and the mass percentage is: Carvacrol 0.1% to 30%, hinokitiol 0.1% to 30%, emulsifier 5% to 10%, dispersant 1% to 10%, antifreeze 0.5% to 5%, preservative 0.5% to 5%, solvent 5% to 15%, water to make up to 100%; The dispersant is polyvinyl alcohol, gum arabic, xanthan gum or magnesium aluminum silicate; The solvent is at least one of sec-butyl acetate, isoamyl citrate, ethyl acetate, cyclohexanone, furfural, 5-methylfurfural, ethanol and ethyl acetate; The antifreeze agent is ethylene glycol, propylene glycol, glycerin or urea; The preservative is sorbic acid, benzoic acid or benzaldehyde; The emulsifier is at least one of alkyl polyoxyethylene ether, fatty acid polyoxyethylene ester, TX-10, 7781T, 4103, OP-10, Well201, Well205 and M30.

9. The composite fungicide of carvacrol and hinokitiol according to claim 7, characterized in that: The bactericide is a soluble liquid, and the mass percentage is: Carvacrol 0.1% to 30%, hinokitiol 0.1% to 30%, emulsifier 5% to 25%, antifreeze 1% to 5%, and solvent to make up to 100%; The emulsifier is at least one of phenylethylphenol polyoxyethylene ether, TX-10, 2201, Ningru 700, Nongru 500, Nongru 600, Nongru BY, Nongru EL and NP-30; The solvent is at least one of methanol, ethanol, cyclohexanone, ethyl acetate, sec-butyl acetate, furfural and 5-methylfurfural; The antifreeze agent is ethylene glycol, propylene glycol, glycerin or urea.

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