Biological herbicide and preparation method thereof
By mixing plant powder with enzymatic solution and microbial fermentation broth, and adding deep eutectic solvents and nanoclay minerals and other ingredients, an efficient and environmentally friendly biological herbicide was prepared, which solved the problems of poor dispersion, insufficient stability and unstable herbicide effects of existing biological herbicide components, and achieved significant herbicidal effect and environmental protection performance.
Patent Information
- Application Number
- CN202510185270.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-19
- Publication Date
- 2025-05-23
AI Technical Summary
In actual application, existing biological herbicides have problems such as poor dispersion of components, insufficient stability, and unstable herbicide effect.
A biological herbicide preparation method is adopted. By mixing plant powders such as mugwort, marshmallow, mugwort, cassia, etc. with pure water, acid buffer and complex enzyme, after enzymatic treatment, it is mixed with deep eutectic solvent and microbial fermentation broth, microwave treatment and ultrasonic demulsification, and finally nanoclay minerals, biostimulators, dispersants and stabilizers to form biological herbicides.
The biological herbicide prepared by this method showed significant herbicidal effect in field experiments. The number of weeds in the number of weeds can reach more than 90% within 3 days after application, and the effect can reach more than 95% after 7 days. After 14 days, the weeds are completely inhibited and do not contain chemically synthesized toxic and harmful substances, which meets the development needs of green agriculture.
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Abstract
Description
[0001] The invention relates to the field of pesticides, and in particular to a method for preparing a biological herbicide. Background Art
[0002] The present invention relates to a biological herbicide and a preparation method thereof, and belongs to the field of agricultural biotechnology. With the rapid development of global agriculture, the negative impact of weeds on crop growth is becoming increasingly significant. The long-term use of traditional chemical herbicides not only causes environmental pollution, but also may cause weed resistance problems, posing a threat to ecological balance and sustainable agricultural development. Therefore, the development of efficient, environmentally friendly and sustainable biological herbicides has become an important research direction of modern agricultural technology.
[0003] In recent years, bioherbicides have attracted widespread attention due to their environmental friendliness, safety to non-target organisms, and renewability. Bioherbicides mainly use natural plant extracts, microbial metabolites, and biostimulants to inhibit weed growth and promote the healthy growth of crops.
[0004] However, existing bioherbicides still face some challenges in practical applications, such as poor dispersion of ingredients, insufficient stability, and unstable weed control effects. Summary of the invention
[0005] In view of the above problems, the present invention provides a method for preparing a biological herbicide.
[0006] To achieve the above objectives, the present invention is implemented through the following technical solutions:
[0007] A method for preparing a biological herbicide comprises the following steps:
[0008] S1: Dry mugwort, marigold, mugwort and cassia seed, then grind them through a 60-mesh sieve to obtain plant powder, add the Asteraceae plant powder into purified water, add an acidic buffer pH regulator and a complex enzyme, adjust the pH to 5-5.5, stir at a constant temperature of 40°C for 2-3h, inactivate the enzyme at 80°C for 10min, centrifuge at high speed for 15-20min, discard the supernatant and retain the precipitate to obtain an enzymatic precipitate;
[0009] S2: Choline chloride and lactic acid are mixed in proportion, stirred at 80°C until a transparent liquid is formed, deionized water is added to reduce the viscosity, and a deep eutectic solvent is obtained for use, the enzymatic hydrolysis precipitate and the deep eutectic solvent are mixed, and transferred to a microwave reactor, and the microwave power is set to 500W, the temperature is 60°C to 70°C, and the time is 15 to 20min. After the reaction, it is quickly cooled to room temperature, and ultrasonic-assisted demulsification is performed at 40kHz for 10 to 15min. After ultrasonic-assisted demulsification, high-speed centrifugation is performed for 20 to 25min, and the supernatant is taken and rotary evaporated to remove the deep eutectic solvent to obtain a plant extract;
[0010] S3: Cultivating Paecilomyces lilacinus in a fermentation tank for 72 to 96 hours, controlling the fermentation temperature at 28°C to 30°C, pH 6.5 to 7.0, aeration volume 1.0 vvm (volume / volume minute), stirring speed 200 rpm, and after the fermentation is completed, removing the bacteria by high-speed centrifugation, and retaining the supernatant as the microbial fermentation liquid;
[0011] S4: mixing the plant extract prepared in S2 and the microbial fermentation liquid prepared in S3, adding nano clay minerals, biostimulants, dispersants and stabilizers, and stirring evenly to obtain a bio-herbicide.
[0012] The CFU number of Paecilomyces lilacinus in the microbial fermentation broth is 10 7 ~10 8 Pieces / ml.
[0013] The terpenoid substances in the Asteraceae plants of Artemisia argyi, Tagetes erecta and Artemisia argyi have the function of inhibiting the germination of weed seeds and the growth of seedlings, and can hinder the growth and development of weeds by affecting the hormone balance in the weeds.
[0014] The phenolic acid substances extracted from the leguminous plant Cassia seed, such as protocatechuic acid and vanillic acid, can inhibit the photosynthesis and respiration of weeds, thereby inhibiting the growth of weeds.
[0015] The humic acid and fulvic acid both contain a variety of active groups, such as carboxyl, phenolic hydroxyl, carbonyl, etc., which can form stable chemical bonds or intermolecular forces with the components in the plant extract, such as hydrogen bonds, chelation, etc. Biostimulants can interact with the metabolites of Paecilomyces lilacinus to enhance its regulatory effect on weed growth; biostimulants can also promote the absorption of nutrients by plants, improve the stress resistance of plants, and indirectly enhance the weed control effect. Humic acid and fulvic acid serve as a bridge connecting plant extracts and microbial fermentation broth, enhancing the synergistic effect between the two; humic acid and fulvic acid improve the overall performance of biological herbicides, so that while removing weeds, they have a certain promoting effect on the growth of crops.
[0016] The sodium lignin sulfonate is an anionic surfactant having hydrophilic and hydrophobic groups. In the biological herbicide system, it can reduce the surface tension of the liquid, so that the plant extract and the microbial fermentation liquid are better dispersed in the solvent. At the same time, it may form a micellar structure with the components in the plant extract, encapsulating the insoluble or poorly soluble components therein, thereby improving their solubility in the solution. When mixed with nano-clay minerals, sodium lignin sulfonate can be adsorbed on the surface of nano-particles, preventing the agglomeration of nano-particles, and further improving the dispersibility and stability of the nano-clay minerals.
[0017] Preferably, the mass proportions of the wormwood, marigold, mugwort, cassia seed, purified water, complex enzyme, choline chloride, lactic acid, and deionized water are 20-30 parts, 16-20 parts, 20-24 parts, 10-15 parts, 150-200 parts, 0.5-0.1 parts, 10-12 parts, 10-12 parts, and 10-15 parts.
[0018] Preferably, the mass proportions of the plant extract, microbial fermentation broth, nano-clay mineral, biostimulant, dispersant and stabilizer are 10-15 parts, 30-40 parts, 2-5 parts, 1-2 parts, 0.5-1 parts, 0.5-1 parts.
[0019] Preferably, the acidic buffer pH regulator is a citric acid-sodium citrate buffer.
[0020] Preferably, the complex enzyme is composed of cellulose, hemicellulose and pectin, and the mass ratio of cellulose, hemicellulose and pectin is 2:1:1.
[0021] Preferably, the nano clay mineral is nano montmorillonite.
[0022] Preferably, the biostimulant consists of humic acid and fulvic acid, and the mass ratio of humic acid to fulvic acid is 1:1.
[0023] Preferably, the dispersant is sodium lignin sulfonate.
[0024] Preferably, the stabilizer is gum arabic.
[0025] Beneficial Effects
[0026] The present invention provides a method for preparing a biological herbicide, which has the following beneficial effects:
[0027] The bio-herbicide combines a variety of natural plant extracts (such as mugwort, marigold, mugwort, cassia seed) and microbial fermentation liquid (Paecilomyces lilacinus). These ingredients work synergistically to effectively inhibit the growth of a variety of common weeds. Field tests have shown that the weed control effect can reach more than 90% within 3 days after application, more than 95% after 7 days, and the weeds are completely suppressed after 14 days, showing a significant weed control effect.
[0028] 1. Compared with traditional chemical herbicides, this biological herbicide is composed entirely of natural plant extracts, microbial fermentation liquid and natural minerals, and does not contain any chemically synthesized toxic and harmful substances. During use, it will not pollute the soil, water sources and air, nor will it have a negative impact on non-target organisms (such as beneficial insects, birds, etc.), which meets the development needs of modern green agriculture. DETAILED DESCRIPTION
[0029] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. 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.
[0030] Embodiment 1:
[0031] A method for preparing a biological herbicide comprises the following steps:
[0032] S1: Take 25 parts of mugwort, 18 parts of longevity flower, 22 parts of mugwort, and 12 parts of cassia seed by mass, dry and grind through a 60-mesh sieve to obtain plant powder. Add the plant powder to 160 parts of purified water, add citric acid-sodium citrate buffer to adjust the pH to 5.3, and add 0.8 parts of a complex enzyme composed of cellulose, hemicellulose and pectin in a ratio of 2:1:1. After constant temperature stirring at 40°C for 2.5h, inactivate the enzyme at 80°C for 10min, centrifuge at high speed for 18min, discard the supernatant and retain the precipitate to obtain the enzymatic precipitate.
[0033] S2: 11 parts of choline chloride and 11 parts of lactic acid were mixed, stirred at 80°C to form a transparent liquid, 12 parts of deionized water were added to reduce the viscosity to form a deep eutectic solvent, the enzymatic precipitate was mixed with the deep eutectic solvent, and transferred to a microwave reactor, and the microwave power was set to 500W, the temperature was 65°C, and the time was 18min. After the end, it was cooled to room temperature, ultrasonic-assisted demulsification (40kHz, 12min), and then high-speed centrifugation was performed for 22min, and the supernatant was taken and rotary evaporated to remove the deep eutectic solvent to obtain a plant extract.
[0034] S3: Cultivate Paecilomyces lilacinus in a fermentation tank for 72 hours, control the fermentation temperature at 29°C, pH 6.8, ventilation volume at 1.0 vvm, and stirring speed at 200 rpm. After the fermentation is completed, remove the bacteria by high-speed centrifugation, and retain the supernatant as the microbial fermentation liquid.
[0035] S4: 12 parts of plant extract and 35 parts of microbial fermentation liquid were mixed, and 3 parts of nano-montmorillonite, 1.5 parts of biostimulant composed of humic acid and fulvic acid in a ratio of 1:1, 0.8 parts of sodium lignin sulfonate, and 0.7 parts of gum arabic were added, and the mixture was stirred evenly to obtain a biological herbicide.
[0036] Embodiment 2:
[0037] A method for preparing a biological herbicide comprises the following steps:
[0038] S1: Weigh 20 parts of mugwort, 16 parts of longevity flower, 20 parts of mugwort, and 10 parts of cassia seed, dry and crush them through a 60-mesh sieve to obtain plant powder. Add them to 150 parts of purified water, adjust the pH to 5.2 with citric acid-sodium citrate buffer, and add 0.6 parts of a composite enzyme with a mass ratio of cellulose, hemicellulose and pectin of 2:1:1. After constant temperature stirring at 40℃ for 2h, inactivate the enzyme at 80℃ for 10min, centrifuge at high speed for 16min, discard the supernatant and retain the precipitate to obtain the enzymatic precipitate.
[0039] S2: Mix 10 parts of choline chloride and 10 parts of lactic acid, stir at 80°C to form a transparent liquid, and then add 10 parts of deionized water to reduce the viscosity to obtain a deep eutectic solvent. Mix the enzymatic precipitate and the deep eutectic solvent, and treat them in a microwave reactor at 500W power and 62°C for 16 minutes. After the end, cool to room temperature, ultrasonically demulsify (40kHz, 11 minutes), centrifuge at high speed for 21 minutes, and take the supernatant and rotary evaporate to remove the deep eutectic solvent to obtain a plant extract.
[0040] S3: Paecilomyces lilacinus was cultured in a fermentation tank for 84 hours at a fermentation temperature of 28°C, pH 6.6, aeration volume of 1.0 vvm, and stirring speed of 200 rpm. After the fermentation was completed, high-speed centrifugation was performed to retain the supernatant, which was the microbial fermentation liquid.
[0041] S4: Mix 10 parts of plant extract with 32 parts of microbial fermentation liquid, add 2.5 parts of nano-montmorillonite, 1.2 parts of biostimulant (humic acid and fulvic acid mass ratio of 1:1), 0.6 parts of sodium lignin sulfonate, and 0.5 parts of gum arabic, and stir well to prepare a biological herbicide.
[0042] Embodiment 3:
[0043] A method for preparing a biological herbicide comprises the following steps:
[0044] S1: Take 30 parts of mugwort, 20 parts of longevity flower, 24 parts of mugwort, and 15 parts of cassia seed, dry and grind them through a 60-mesh sieve to obtain plant powder. Put them into 200 parts of pure water, adjust the pH to 5.4 with citric acid-sodium citrate buffer, add 11 parts of complex enzyme with a mass ratio of cellulose, hemicellulose and pectin of 2:1:1, stir at a constant temperature of 40°C for 3h, inactivate the enzyme at 80°C for 10min, centrifuge at high speed for 20min, discard the supernatant and retain the precipitate to obtain the enzymatic precipitate.
[0045] S2: 12 parts of choline chloride and 12 parts of lactic acid were mixed, stirred at 80°C to form a transparent liquid, and then 15 parts of deionized water were added to reduce the viscosity to obtain a deep eutectic solvent. The enzymatic precipitate was mixed with the deep eutectic solvent, and treated in a microwave reactor at 500 W power and 68°C for 15 minutes. After the treatment, it was cooled to room temperature, ultrasonically demulsified (40kHz, 13 minutes), and centrifuged at high speed for 24 minutes. The supernatant was taken and rotary evaporated to remove the deep eutectic solvent to obtain a plant extract.
[0046] S3: Paecilomyces lilacinus was cultured in a fermenter for 96 hours at a fermentation temperature of 30°C, pH 7.0, aeration volume of 1.0 vvm, and stirring speed of 200 rpm. After the fermentation was completed, high-speed centrifugation was performed and the supernatant was retained as the microbial fermentation liquid.
[0047] S4: Mix 15 parts of plant extract with 38 parts of microbial fermentation liquid, add 4 parts of nano-montmorillonite, 2 parts of biostimulant (humic acid and fulvic acid mass ratio of 1:1), 1 part of sodium lignin sulfonate, and 1.2 parts of gum arabic, and stir well to prepare a biological herbicide.
[0048] Embodiment 4:
[0049] A method for preparing a biological herbicide comprises the following steps:
[0050] S1: Weigh 28 parts of mugwort, 19 parts of longevity flower, 23 parts of mugwort, and 14 parts of cassia seed, dry and grind them through a 60-mesh sieve to obtain plant powder. Place them in 170 parts of pure water, adjust the pH to 5.5 with citric acid-sodium citrate buffer, and add 0.9 parts of a complex enzyme with a ratio of cellulose, hemicellulose and pectin of 2:1:1. After constant temperature stirring at 40℃ for 2.8h, inactivate the enzyme at 80℃ for 10min, centrifuge at high speed for 19min, discard the supernatant and retain the precipitate to obtain the enzymatic precipitate.
[0051] S2: 11.5 parts of choline chloride and 11.5 parts of lactic acid were mixed, stirred at 80°C to form a transparent liquid, and then 12 parts of deionized water were added to reduce the viscosity to obtain a deep eutectic solution. The enzymatic hydrolysis precipitate was mixed with the deep eutectic solvent, and treated in a microwave reactor at a power of 500 W and a temperature of 67°C for 19 minutes. After the treatment, it was cooled to room temperature, ultrasonic-assisted demulsification (40 kHz, 14 minutes), and high-speed centrifugation was performed for 23 minutes. The supernatant was taken and rotary evaporated to remove the deep eutectic solvent to obtain a plant extract.
[0052] S3: Paecilomyces lilacinus was cultured in a fermentation tank for 80 hours at a fermentation temperature of 28.5°C, pH 6.7, aeration volume of 1.0 vvm, and stirring speed of 200 rpm. After the fermentation was completed, high-speed centrifugation was performed to retain the supernatant, i.e., the microbial fermentation liquid.
[0053] S4: 13 parts of plant extract were mixed with 36 parts of microbial fermentation liquid, 3.5 parts of nano-montmorillonite, 1.8 parts of biostimulant (humic acid and fulvic acid mass ratio of 1:1), 0.9 parts of sodium lignin sulfonate and 0.8 parts of gum arabic were added, and the mixture was stirred evenly to prepare a biological herbicide.
[0054] Embodiment 5:
[0055] A method for preparing a biological herbicide comprises the following steps:
[0056] S1: Take 22 parts of mugwort, 17 parts of longevity flower, 21 parts of mugwort, and 13 parts of cassia seed, dry and grind them through a 60-mesh sieve to obtain plant powder, add them to 150 parts of purified water, adjust the pH to 5.1 with citric acid-sodium citrate buffer, and add 0.7 parts of a composite enzyme with a ratio of cellulose, hemicellulose and pectin of 2:1:1. After constant temperature stirring at 40°C for 2.2 hours, inactivate the enzyme at 80°C for 10 minutes, centrifuge at high speed for 17 minutes, discard the supernatant and retain the precipitate to obtain an enzymatic precipitate.
[0057] S2: 10.5 parts of choline chloride and 10.5 parts of lactic acid were mixed, stirred at 80°C to form a transparent liquid, and then 13 parts of deionized water were added to reduce the viscosity to obtain a deep eutectic solvent. The enzymatic precipitate was mixed with the deep eutectic solvent and treated in a microwave reactor at 500W power and 63°C for 17 minutes. After the end, it was cooled to room temperature, ultrasonic-assisted demulsification (40kHz, 10min), high-speed centrifugation for 20min, and the supernatant was rotary evaporated to remove the deep eutectic solvent to obtain a plant extract.
[0058] S3: Paecilomyces lilacinus was cultured in a fermenter for 78 hours at a fermentation temperature of 29.5°C, pH 6.9, aeration volume of 1.0 vvm, and stirring speed of 200 rpm. After the fermentation was completed, high-speed centrifugation was performed to retain the supernatant, i.e., the microbial fermentation liquid.
[0059] S4: Mix 11 parts of plant extract with 33 parts of microbial fermentation liquid, add 3.2 parts of nano-montmorillonite, 1.6 parts of biostimulant (humic acid and fulvic acid mass ratio of 1:1), 0.7 parts of sodium lignin sulfonate, and 0.6 parts of gum arabic, and stir well to prepare a biological herbicide.
[0060] Comparative Example 1:
[0061] The deep eutectic solvent in step S1 was replaced with 70% ethanol aqueous solution, and the remaining raw material ratios, preparation methods, and operating conditions were the same as those in Example 1.
[0062] Comparative Example 2:
[0063] No biostimulant was added (the mass ratio of humic acid to fulvic acid was 1:1), and the remaining raw material ratios, preparation methods, and operating conditions were the same as those in Example 1.
[0064] Comparative Example 3:
[0065] No nano clay mineral was added, and the remaining raw material ratios, preparation methods, and operating conditions were the same as those in Example 1.
[0066] Field efficacy test
[0067] Barnyard grass, crabgrass, Amaranthus retroflexus, Erigeron cantoniensis, Artemisia chrysanthemum, Cyperus rotundus, and Chenopodium album were sown in the field. When the weeds grew to the 3-5 leaf stage, the biological herbicides prepared in Examples 1-5 and Comparative Examples 1-3 were sprayed (repeated 3 times in each area). The number of weeds was investigated 3d, 7d, and 14d after the application, and the control effect per plant was calculated: The weed control effect is shown in Table 1:
[0068] Table 1
[0069]
[0070]
[0071] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the equivalents of the appended claims.
Claims
1. A method for preparing a biological herbicide, characterized in that ： A method for preparing a biological herbicide comprises the following steps: S1: Dry mugwort, marigold, mugwort and cassia seed, then grind them through a 60-mesh sieve to obtain plant powder, add the Asteraceae plant powder into purified water, add an acidic buffer pH regulator and a complex enzyme, adjust the pH to 5-5.5, stir at a constant temperature of 40°C for 2-3h, inactivate the enzyme at 80°C for 10min, centrifuge at high speed for 15-20min, discard the supernatant and retain the precipitate to obtain an enzymatic precipitate; S2: Choline chloride and lactic acid are mixed in proportion, stirred at 80°C until a transparent liquid is formed, deionized water is added to reduce the viscosity, and a deep eutectic solvent is obtained for use, the enzymatic hydrolysis precipitate and the deep eutectic solvent are mixed, and transferred to a microwave reactor, and the microwave power is set to 500W, the temperature is 60°C to 70°C, and the time is 15 to 20min. After the reaction, it is quickly cooled to room temperature, and ultrasonic-assisted demulsification is performed at 40kHz for 10 to 15min. After ultrasonic-assisted demulsification, high-speed centrifugation is performed for 20 to 25min, and the supernatant is taken and rotary evaporated to remove the deep eutectic solvent to obtain a plant extract; S3: Cultivating Paecilomyces lilacinus in a fermentation tank for 72 to 96 hours, controlling the fermentation temperature at 28°C to 30°C, pH 6.5 to 7.0, aeration volume 1.0 vvm (volume / volume minute), stirring speed 200 rpm, and after the fermentation is completed, removing the bacteria by high-speed centrifugation, and retaining the supernatant as the microbial fermentation liquid; S4: mixing the plant extract prepared in S2 and the microbial fermentation liquid prepared in S3, adding nano clay minerals, biostimulants, dispersants and stabilizers, and stirring evenly to obtain a bio-herbicide.
2. The method for preparing a biological herbicide according to claim 1, characterized in that: The acidic buffer pH regulator is a citric acid-sodium citrate buffer.
3. The method for preparing a biological herbicide according to claim 1, characterized in that: The complex enzyme consists of cellulose, hemicellulose and pectin, and the mass ratio of cellulose, hemicellulose and pectin is 2:1:
1.
4. The method for preparing a biological herbicide according to claim 1, characterized in that: The nano clay mineral is nano montmorillonite.
5. The method for preparing a biological herbicide according to claim 1, characterized in that: The biostimulant consists of humic acid and fulvic acid, and the mass ratio of humic acid to fulvic acid is 1:
1.
6. The method for preparing a biological herbicide according to claim 1, characterized in that: The dispersant is sodium lignin sulfonate.
7. The method for preparing a biological herbicide according to claim 1, characterized in that: The stabilizer is gum arabic.
8. A biological herbicide, characterized in that: The herbicide is obtained by the preparation method of the biological herbicide according to any one of claims 1 to 7.