Specialized bio-enzyme insecticide for leaf-eating grasses, its preparation method and application
Patent Information
- Application Number
- CN202610928717.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-06-25
- Publication Date
- 2026-09-15
Abstract
Description
Technical Field
[0001] This invention relates to the field of agricultural biological insecticides, specifically to a biological enzyme insecticide for leafy grasses, its preparation method, and its application. Background Technology
[0002] Edible leaf grass (also known as protein grass) is a perennial herbaceous plant used for both medicinal and edible purposes, as well as forage and food. It has unique growth characteristics such as salt and alkali tolerance, cold and heat resistance, tolerance to poor soil, well-developed root system, and multiple harvests. Its leaves are rich in nutrients such as protein and amino acids, and it is widely used in food, feed, and deep processing, making it of high economic value.
[0003] However, leafy greens are susceptible to various leaf pests during cultivation, mainly including cabbage caterpillars, diamondback moths, aphids, leaf miners, and flea beetles. These pests gnaw on the leaves and damage the leaf tissue, causing yellowing and damage to the leaves. This not only reduces the yield and quality of the leafy greens but also affects their subsequent edible, feed, and processing value. Furthermore, due to the special status of leafy greens as both food and medicine, the safety requirements for pest control products are extremely high. The use of pesticides containing harmful ingredients such as chemical pesticides, azadirachtin, and chemical toxins is strictly prohibited to avoid residues that could harm humans, livestock, and the ecological environment.
[0004] Currently, agricultural pesticides are mainly divided into two categories: chemical pesticides and biological pesticides. (1) Chemical insecticides: Although they kill insects quickly, they have problems such as high residue, environmental pollution, and easy development of insect resistance in pests. They are not suitable for planting leafy grasses.
[0005] (2) Existing biological insecticides: Most are general-purpose products and are not specifically designed for the growth characteristics of leafy grasses, such as salt and alkali tolerance, strong adaptability to high and low temperatures, and multiple harvests. They have the following technical defects: Poor adaptability: It has a narrow pH range and cannot adapt to the slightly alkaline soil environment preferred by leafy grasses; its enzyme activity is unstable under high and low temperature conditions, affecting the planting effect in both northern and southern regions. Poor leaf protection effect: Due to the lack of leaf nutrition replenishment and damage repair function, the leaves are still prone to yellowing and damage after pest control; Single function: It cannot meet the needs of perennial herbaceous plant cultivation, such as soil improvement and root care. Safety concerns: Some products still contain small amounts of plant pesticides or chemical additives, which does not meet the safety positioning of edible leaves as "food and medicine of the same origin" with no residue and no toxins. Large-scale production is difficult: the formula and process are complex and prone to foaming, layering and sedimentation, making it difficult to meet the needs of factory mass production and commercial supply.
[0006] Therefore, developing a specialized bio-enzyme insecticide that is adapted to the growth characteristics of leafy grasses, is free of pesticide residues and toxins, has stable insecticidal effects, and also has the functions of protecting leaves and improving soil, and can be mass-produced, has become an urgent technical problem to be solved. Summary of the Invention
[0007] In order to solve the problems of the prior art, the present invention provides a biological enzyme insecticide for leafy grass, its preparation method and application.
[0008] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution: Firstly, a biological enzyme insecticide specifically for leafy grasses, based on 1000 kg of finished product, is composed of the following raw materials in parts by weight: 140-150 parts of compound enzyme, 60-68 parts of auxiliary agent, and 782-800 parts of commercial-grade dechlorinated purified water; The complex enzyme assembly includes chitinase, neutral protease, amylase, lipase, and polypeptide enzyme; wherein, the chitinase activity is ≥6500U / g, the neutral protease activity is ≥28000U / g, the amylase activity is ≥12000U / g, the lipase activity is ≥9000U / g, and the polypeptide enzyme activity is ≥18000U / g. The adjuvant assembly includes a penetration system, an antibacterial and repellent system, an enzyme stabilizing system, a pH adjuster, leaf-protecting and synergistic ingredients, and commercial auxiliary ingredients; The osmotic system is composed of tea saponin and natural trehalose; The antibacterial and repellent system is composed of a compound plant extract of Sophora flavescens, lemongrass, and peppermint, licorice extract, and artemisia argyi extract. The enzyme stabilizing system is composed of sorbitol, xanthan gum, and natural plant moisturizing factors. The pH adjuster is composed of citric acid, sodium bicarbonate, and a natural buffer. The leaf-protecting and synergistic ingredient is a natural plant amino acid; The commercial auxiliary ingredients include food-grade defoamers and natural biological preservatives.
[0009] In one specific embodiment of the first aspect, based on 1000 kg of finished product, the total amount of the compound enzyme is 145 kg, including 42 kg of chitinase, 50 kg of neutral protease, 18 kg of amylase, 13 kg of lipase, and 22 kg of peptidase; the total amount of the adjuvants is 63.5 kg, including 28 kg of osmotic system, 24 kg of antibacterial and repellent system, 7 kg of enzyme stabilizing system, 3 kg of pH adjuster, 1 kg of leaf-protecting and synergistic ingredient, and 0.5 kg of commercial auxiliary ingredients; and the commercial-grade dechlorinated purified water is 790 kg.
[0010] In one specific embodiment of the first aspect, the permeation system consists of 24 kg of commercial-grade tea saponin and 4 kg of commercial-grade natural trehalose; the antibacterial and repellent system consists of 16 kg of commercial-grade Sophora flavescens-citronella-peppermint compound plant extract, 5 kg of commercial-grade licorice extract, and 3 kg of commercial-grade Artemisia argyi extract; the enzyme stabilizing system consists of 4 kg of commercial-grade sorbitol, 2 kg of commercial-grade xanthan gum, and 1 kg of commercial-grade natural plant moisturizing factor; the pH adjuster consists of 1.5 kg of commercial-grade citric acid, 0.5 kg of commercial-grade sodium bicarbonate, and 1 kg of commercial-grade natural buffer; the leaf-protecting synergistic ingredient is 1 kg of commercial-grade natural plant amino acids; and the commercial auxiliary ingredients consist of 0.3 kg of food-grade defoamer and 0.2 kg of commercial-grade natural biological preservative.
[0011] In one specific embodiment of the first aspect, the chitinase, neutral protease, amylase, lipase, and polypeptide enzyme are all commercially available bulk-purchased raw materials; the tea saponin, trehalose, plant extracts, licorice extract, artemisia extract, sorbitol, xanthan gum, natural plant moisturizing factor, natural buffer, natural plant amino acids, defoamer, and natural biological preservative are all food-grade or agricultural biological-grade, free of alcohol, chemical preservatives, strong acid and strong alkali components, and biodegradable.
[0012] In one specific embodiment of the first aspect, the natural buffer can adjust the pH value of the insecticide to 5.5-8.0; the natural plant moisturizing factor can enhance the activity stability of the enzyme under high and low temperature environments; and the natural biological preservative can extend the shelf life of the finished product to 18 months.
[0013] Secondly, a method for preparing a bio-enzyme insecticide specifically for leafy grasses includes the following steps: Step 1: Pre-production preparation, sampling and testing of raw materials, cleaning, disinfecting and drying of production equipment, and adjusting the temperature of the production workshop to 15℃-25℃; Step 2: Dechlorination treatment. Treat tap water to dechlorinated purified water, test the pH value to 5.5-7.5, pour it into a mixing tank, and add 70%-85% of the total amount of dechlorinated purified water. Step 3: Enzyme solution preparation. Turn on the mixing tank and stir at medium speed. Add chitinase, neutral protease, amylase, lipase and peptidase in sequence. Stir until dissolved evenly after each addition. Step 4: Add penetrating ingredients, including tea saponin and natural trehalose, along with an antifoaming agent, and stir well; Step 5: Add antibacterial and repellent ingredients. Add the compound plant extracts of Sophora flavescens, lemongrass, and peppermint, licorice extract, and artemisia argyi extract, and stir well. Step 6: Add enzyme-stabilizing and leaf-protecting ingredients, including sorbitol, xanthan gum, natural plant moisturizing factors, and natural plant amino acids, and stir until completely dissolved; Step 7: Precisely adjust the pH by slowly adding citric acid, sodium bicarbonate, and natural buffer while stirring and measuring the pH value to stabilize it between 5.5 and 8.0. Step 8: Adjust volume and homogenize, add the remaining dechlorinated purified water to the total volume, increase the stirring speed, and homogenize; Step 9: Defoaming and filtration. Let the mixture stand to defoam naturally, then filter it through a precision filtration device. Step 10: Filling and sealing. Fill to commercial specifications and seal immediately. Step 11: Finished product storage. Place the finished products in a cool, dry, and well-ventilated warehouse, with the temperature controlled between 5℃ and 30℃. Arrange them by batch and label them properly.
[0014] In one specific embodiment of the second aspect, the dechlorinated purified water added in step 2 accounts for 80% of the total volume; in step 3, the stirring speed is 60-80 r / min, and each enzyme is added and stirred for 5 minutes; in step 8, after replenishing the dechlorinated purified water, the speed is adjusted to 80-100 r / min, and homogenized stirring is performed for 15 minutes; in step 9, the mixture is allowed to stand for 30 minutes to naturally defoam, and then filtered through a 200-mesh precision filter.
[0015] In one specific implementation of the second aspect, the method can scale batch sizes proportionally to adapt to production in factories of different sizes.
[0016] Thirdly, an application method for a bio-enzyme insecticide specifically for leafy grasses, used for pest control at different growth stages of leafy grasses, specifically including: (1) Seedling stage prevention: Dilute with water at a ratio of 1:200 to 1:300 and spray on both sides of the leaves once every 5 to 10 days; (2) Prevention and control during the growing season: For routine prevention, dilute with water at a ratio of 1:150 to 1:250. During the peak period of pests, dilute with water at a ratio of 1:30 to 1:100. Repeat every 3 to 7 days for 2 to 3 consecutive times. (3) Post-harvest recovery period: Dilute with water at a ratio of 1:150 to 1:250 and spray on new leaves and roots once every 7 to 14 days; (4) Control of underground pests: Dilute with water at a ratio of 1:250 to 1:500 and apply as a root irrigation treatment. Use 600-800g of the original solution per mu after diluting with water.
[0017] In one specific implementation of the third aspect, seedling prevention is diluted with water at a ratio of 1:250 and applied once every 7 days; routine prevention during the growing season is diluted with water at a ratio of 1:200; during periods of high pest incidence, the solution is diluted with water at a ratio of 1:50 to 1:80 and applied once every 3-5 days; during the post-harvest recovery period, the solution is diluted with water at a ratio of 1:200 and applied once every 10 days; and underground pest control is diluted with water at a ratio of 1:300 to 1:400. The insecticide dilution solution should be prepared and used immediately, and used up within 24 hours; it should not be mixed with strong acid or strong alkali pesticides or fungicides, but can be mixed with organic fertilizers, biological agents, and neutral foliar fertilizers; use should be stopped 3 days before the harvest of leafy grasses.
[0018] The beneficial effects of this invention are as follows: 1. Highly targeted, with stable and efficient insecticidal effects; This invention targets common leaf pests (cabbage caterpillars, diamondback moths, aphids, leaf miners, flea beetles) and underground nematodes prevalent in leaf-eating grasses, using a high-activity compound bio-enzyme (chitinase ≥6500U / g, neutral protease ≥28000U / g, amylase ≥12000U / g, lipase ≥9000U / g, and peptidase ≥18000U / g). Chitinase and neutral protease work synergistically to rapidly degrade chitin and proteins in the insect's body wall and egg shell, disrupting its physical barrier; amylase and lipase interfere with the insect's energy metabolism, while peptidase enhances plant immunity. Example 2 verified that the incidence of leaf pests decreased to 3.2%, the egg inactivation rate reached 88.5%, and the underground nematode incidence rate was only 2.8%, significantly superior to commercially available general-purpose biological insecticides (infestation rate 12.5%, egg inactivation rate 65.3%). The onset time is consistently between 2 and 30 hours, meeting the rapid prevention and control needs of large-scale planting.
[0019] 2. Excellent adaptability, perfectly matching the growth characteristics of leafy plants. This invention achieves targeted adaptation through the following design: Salt and alkali tolerance adaptation: The pH adjuster stabilizes the pH of the finished product at 5.5-8.0, covering the slightly alkaline soil environment preferred by leafy grasses, without affecting their salt and alkali tolerance growth characteristics; Temperature adaptability: The synergistic effect of sorbitol, xanthan gum and natural plant moisturizing factors in the enzyme-stabilizing system results in an enzyme activity retention rate of ≥85% in an environment of 5℃~40℃, making it suitable for open-air and greenhouse cultivation of leafy grass in both northern and southern regions. Multiple harvesting adaptation: Example 2 shows that after using the present invention, the new leaf germination time after harvest is only 3.5 days (6.2 days in the control group), which significantly shortens the recovery period and fits the planting pattern of multiple harvests of leafy grass in one year; Root care: Polypeptide enzymes and Artemisia argyi extract promote root development. Root irrigation can effectively kill underground nematodes, protect the perennial roots, and help the long-term planting of leafy grass (one planting can be harvested continuously for 10-15 years).
[0020] 3. Safe and compliant, with no residue risk. All raw materials used in this invention are food-grade or agricultural biological grade, completely eliminating harmful components such as chemical pesticides, azadirachtin, and plant toxins. Residue testing in Example 2 showed that the leafy greens had no pesticide residues or toxins after harvesting, meeting the requirements of "medicinal and edible homology" and dual use as feed / food. The product is biodegradable, does not pollute soil or water sources, and is safe for non-target organisms such as bees and earthworms, meeting the requirements of green organic farming.
[0021] 4. Multifunctional, achieving "insect control + leaf protection + root nourishment + soil improvement" in one; leaf protection and enhanced efficacy: natural plant amino acids directly supplement leaf nutrition, reducing leaf yellowing and damage caused by pests. In Example 2, the leaf integrity of the experimental group reached 98.3% (compared to 82.7% in the control group). Antibacterial and repellent: The compound plant extracts of Sophora flavescens, lemongrass, and peppermint, along with licorice extract and artemisia argyi extract, work together to repel pests and inhibit leaf pathogens, reducing secondary diseases; Soil improvement: Polypeptide enzymes decompose organic matter in the soil, releasing amino acids and small peptides, thereby improving soil fertility. Example 2 shows that the soil fertility of the experimental group was slightly improved, while the control group showed no change. Root care: Root irrigation can kill underground nematodes, while natural trehalose promotes root cell repair and enhances plant resistance.
[0022] 5. Low resistance and stable long-term efficacy: This invention utilizes enzymatic hydrolysis as a physical insecticidal principle, without neurotoxins or metabolic inhibitors, preventing pests from developing resistance. Accelerated stability testing (25℃±2℃, 45%±10% relative humidity, stored in the dark for 6 months) showed an enzyme activity retention rate of ≥85%, with no stratification or precipitation. Suitable for perennial edible grasses, it eliminates the need for frequent product replacement and provides stable long-term efficacy.
[0023] 6. Adaptable to large-scale production with controllable costs; Commercial raw materials: All enzymes and auxiliaries are commercially available, ensuring batch stability and convenient procurement; Standardized processes: Clear preparation methods and steps, with a wide adjustable parameter range (e.g., pH 5.5–8.0, rotation speed 60–100 r / min, temperature 15℃–25℃), adaptable to equipment in factories of different sizes; High efficiency: Defoamer and 200-mesh filter design prevent foaming and clogging, resulting in high filling efficiency and no stratification or sedimentation issues; Cost optimization: Optimized auxiliary ratios without affecting efficacy and safety, resulting in large-scale production costs that are more than 20% lower than similar biological pesticides; Adjustable batch sizes: Batch sizes can be scaled up proportionally (e.g., 500kg, 2000kg, 5000kg) to meet commercial distribution and long-term supply needs.
[0024] 7. Flexible application methods, adaptable to the entire growth cycle; This invention provides differentiated dilution ratios and application schemes for the seedling stage, growth stage (regular and high-incidence periods), post-harvest recovery period, and underground pest control, and clarifies the contraindications (do not mix with strong acid or strong alkali pesticides, discontinue use 3 days before harvest), is easy to operate, suitable for large-scale mechanized spraying or drip irrigation, and has strong practicality. Detailed Implementation
[0025] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.
[0026] A bio-enzyme insecticide specifically for leafy grasses, its preparation method, and its application.
[0027] Example 1: Preparation of a bio-enzyme insecticide specifically for leafy grasses Raw material preparation (based on 1000kg of finished product): Chitinase 42kg (enzyme activity ≥6500U / g, commercial grade), neutral protease 50kg (enzyme activity ≥28000U / g, commercial grade), amylase 18kg (enzyme activity ≥12000U / g, commercial grade), lipase 13kg (enzyme activity ≥9000U / g, commercial grade), peptidase 22kg (enzyme activity ≥18000U / g, commercial grade); Tea saponin 24kg, natural trehalose 4kg, Sophora flavescens-citronella-peppermint compound plant extract 16kg, licorice extract 5kg, Artemisia argyi extract 3kg, sorbitol 4kg, xanthan gum 2kg, natural plant moisturizing factor 1kg, citric acid 1.5kg, sodium bicarbonate 0.5kg, natural buffer 1kg, natural plant amino acids 1kg, food-grade defoamer 0.3kg, natural biological preservative 0.2kg; Replenish commercial-grade dechlorinated purified water to 1000kg.
[0028] Preparation steps: (1) Preparation before production: Sampling and testing all raw materials to ensure that enzyme activity, purity and pH value meet the standards; cleaning and disinfecting 1500L mixing tank, dechlorination water treatment equipment, 200 mesh precision filter equipment and automated filling equipment, and drying them for later use; controlling the temperature of the production workshop at 20℃ and keeping it ventilated and dust-free; operators wearing gloves, masks and protective clothing.
[0029] (2) Dechlorination water treatment: Turn on the dechlorination water treatment equipment to treat the tap water into dechlorinated pure water. The pH value is measured to be 6.0. Pour into the mixing tank and add about 632 kg of dechlorinated pure water (about 80% of the total amount).
[0030] (3) Enzyme solution preparation: Turn on the stirring tank and adjust the speed to 70r / min. Add neutral protease, chitinase, amylase, lipase and peptidase in sequence. Stir for 5 minutes after each enzyme is added. After sampling and testing the enzyme activity, proceed to the next step.
[0031] (4) Adding penetrating ingredients: Add tea saponin and natural trehalose, stir for 8 minutes, and add defoamer at the same time to inhibit foam production.
[0032] (5) Addition of antibacterial and repellent ingredients: Add Sophora flavescens-citronella-mint compound plant extract, licorice extract, and Artemisia argyi extract, and stir for 6 minutes to mix well.
[0033] (6) Adding enzyme-stabilizing and leaf-protecting ingredients: Add sorbitol, xanthan gum, natural plant moisturizing factor and natural plant amino acids, and stir for 10 minutes until completely dissolved.
[0034] (7) Precise pH adjustment: Slowly add citric acid, sodium bicarbonate and natural buffer while stirring and measuring pH. Finally, stabilize the pH value at 6.5 and record the data.
[0035] (8) Volume adjustment and homogenization: Add dechlorinated purified water to 1000 kg, adjust the speed to 90 r / min, and homogenize for 15 minutes to ensure uniform mixing.
[0036] (9) Defoaming and filtration: Let stand for 30 minutes to defoam naturally, then filter through a 200-mesh precision filter. Sampling tests show that the purity of the finished product meets the standards.
[0037] (10) Filling and sealing: Automated filling equipment is used to fill 5L / barrel. The barrel is sealed immediately after filling and the filling accuracy is sampled and tested.
[0038] (11) Finished product storage: Place the finished products in a cool and ventilated warehouse, with the temperature controlled at 20℃, and arrange them by batch and label them properly.
[0039] The enzyme activity detection method for each enzyme component in the complex enzyme assembly used in this embodiment is as follows: (1) Chitinase activity determination: The DNS colorimetric method was used, with colloidal chitin as the substrate, and the reaction was carried out at 40℃ and pH 6.0 for 30 minutes. The amount of enzyme required to produce 1 μmol of reducing sugar (calculated as N-acetylglucosamine) per minute was defined as 1 unit of enzyme activity (U). The chitinase activity used in this example was ≥6500 U / g.
[0040] (2) Neutral protease activity assay: The Folin-Ciocalteu method was used, with casein as the substrate, and the reaction was carried out at 40°C and pH 7.0 for 10 minutes. The amount of enzyme required to hydrolyze and produce 1 μg of tyrosine per minute was defined as 1 enzyme activity unit (U). The neutral protease activity used in this example was ≥28000 U / g.
[0041] (3) Amylase activity determination: The iodine colorimetric method was used with soluble starch as the substrate. The reaction was carried out at 60℃ and pH 6.0 for 5 minutes. The amount of enzyme required to hydrolyze 1 mg of starch per minute was defined as 1 enzyme activity unit (U). The amylase activity used in this example was ≥12000 U / g.
[0042] (4) Lipase activity determination: The titration method was used with olive oil as substrate. The reaction was carried out at 40°C and pH 7.5 for 30 minutes. The amount of enzyme required to release 1 μmol of fatty acid per minute was defined as 1 enzyme activity unit (U). The lipase activity used in this example was ≥9000 U / g.
[0043] (5) Determination of peptidase activity: The ninhydrin colorimetric method was used with glutathione as the substrate. The reaction was carried out at 37°C and pH 7.0 for 10 minutes. The amount of enzyme required to produce 1 μmol of free amino acids per minute was defined as 1 unit of enzyme activity (U). The peptidase activity used in this example was ≥18000 U / g.
[0044] The above detection methods are all conventional methods in this field, and all reagents used are of analytical grade. The detection results are obtained by averaging three replicates.
[0045] Example 2: Application Test of a Special Bio-enzyme Insecticide for Leafy Grasses Experimental site: A large-scale edible leafy grass planting base with an area of 10 mu (approximately 1.65 acres). The soil is saline-alkali land. The planted varieties are conventional edible leafy grasses. The experimental group and the control group are divided into experimental group and control group, with each group having 5 mu (approximately 0.65 acres). The planting conditions (watering, fertilization, and planting density) of the two groups are the same.
[0046] Experimental methods: The experimental group used the leafy grass-specific biological enzyme insecticide prepared in Example 1 of this invention for pest control according to the application method of this invention; the control group used a commercially available general-purpose biological insecticide for pest control according to its instructions.
[0047] The trial period was 3 months, covering the seedling stage, growth stage, and recovery period after the first harvest of leafy grass.
[0048] Experimental results: (1) Pest control effect: The incidence of leaf pests (cabbage caterpillar, diamondback moth, aphid, leaf miner, flea beetle) in the experimental group was 3.2%, the inactivation rate of insect eggs was 88.5%, and the incidence of underground nematodes was 2.8%; the incidence of leaf pests in the control group was 12.5%, the inactivation rate of insect eggs was 65.3%, and the incidence of underground nematodes was 9.6%; the pest control effect of the insecticide of this invention is significantly better than that of commercially available general biological insecticides.
[0049] (2) Leaf damage: The leaves of the experimental group of leafy grass showed no obvious yellowing or damage, and the leaf integrity reached 98.3%; the leaves of the control group showed varying degrees of grazing and yellowing damage, and the leaf integrity was 82.7%; the insecticide of this invention has outstanding leaf protection effect.
[0050] (3) Growth recovery: After the leafy grass in the experimental group was harvested, the new leaves sprouted in 3.5 days, the root system grew well, and the soil fertility improved slightly; the new leaves sprouted in the control group in 6.2 days, the root system grew moderately, and the soil fertility did not change significantly; the insecticide of this invention can effectively help the leafy grass recover after harvesting, and has the functions of soil improvement and root maintenance.
[0051] (4) Residue detection: After the experimental group of leafy grass was harvested, the sample test showed no pesticide residues or toxins, which met the standards for food and feed use; the control group was found to have trace amounts of plant pesticide residues, which did not meet the "medicinal and edible" positioning of leafy grass.
[0052] (5) Stability test: After 6 months of storage, the enzyme activity retention rate of the experimental product was 85%, with no stratification or precipitation, indicating good stability; after 6 months of storage, the enzyme activity retention rate of the control group product was 62%, with slight stratification.
[0053] The experimental results show that the bio-enzyme insecticide of this invention for leafy grass has stable pest control effect, significant leaf protection effect, and is safe and residue-free. It is suitable for the growth characteristics and safety positioning of leafy grass, and is superior to existing general-purpose bio-insecticides. It can be widely used in the large-scale planting of leafy grass.
[0054] In Example 2, the enzyme activity retention rate of the finished product after 6 months of storage was tested under the following specific conditions: The sealed finished product (5L / barrel) was placed in a constant temperature and humidity chamber, with the temperature controlled at 25℃±2℃ and the relative humidity at 45%±10%, avoiding direct sunlight. Samples were taken at 0 months (i.e., the day of preparation) and 6 months after storage. After sampling, the enzyme activities of the five enzymes in the complex enzyme composition were determined according to the corresponding methods in the "Detection Methods" section above. The enzyme activity retention rate was calculated by dividing the enzyme activity after 6 months of storage by the enzyme activity at 0 months and then multiplying by 100%. In this example, the overall enzyme activity retention rate of the finished product after 6 months of storage was 85% (the average retention rate of the five enzymes), and there was no stratification or precipitation, indicating good product stability.
[0055] If you need to test the retention rate at different temperatures, you can refer to the above conditions and set storage conditions such as 5℃ and 30℃ for comparison.
[0056] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.
Claims
1. A bio-enzyme insecticide specifically for leafy grasses, characterized in that, Based on 1000kg of finished product, it consists of the following parts by weight of raw materials: 140-150 parts of compound enzyme, 60-68 parts of auxiliary agent, and 782-800 parts of commercial-grade dechlorinated purified water; The complex enzyme assembly includes chitinase, neutral protease, amylase, lipase, and polypeptide enzyme; wherein, the chitinase activity is ≥6500U / g, the neutral protease activity is ≥28000U / g, the amylase activity is ≥12000U / g, the lipase activity is ≥9000U / g, and the polypeptide enzyme activity is ≥18000U / g. The adjuvant assembly includes a penetration system, an antibacterial and repellent system, an enzyme stabilizing system, a pH adjuster, leaf-protecting and synergistic ingredients, and commercial auxiliary ingredients; The osmotic system is composed of tea saponin and natural trehalose; The antibacterial and repellent system is composed of a compound plant extract of Sophora flavescens, lemongrass, and peppermint, licorice extract, and artemisia argyi extract. The enzyme stabilizing system is composed of sorbitol, xanthan gum, and natural plant moisturizing factors. The pH adjuster is composed of citric acid, sodium bicarbonate, and a natural buffer. The leaf-protecting and synergistic ingredient is a natural plant amino acid; The commercial auxiliary ingredients include food-grade defoamers and natural biological preservatives.
2. The bio-enzyme insecticide for leafy grasses according to claim 1, characterized in that, Based on a finished product of 1000 kg, the total amount of the compound enzyme is 145 kg, including 42 kg of chitinase, 50 kg of neutral protease, 18 kg of amylase, 13 kg of lipase, and 22 kg of peptidase; the total amount of the adjuvants is 63.5 kg, including 28 kg of osmotic system, 24 kg of antibacterial and repellent system, 7 kg of enzyme stabilizing system, 3 kg of pH adjuster, 1 kg of leaf-protecting and synergistic ingredient, and 0.5 kg of commercial auxiliary ingredients; the amount of commercial-grade dechlorinated purified water is 790 kg.
3. The bio-enzyme insecticide for leafy grasses according to claim 2, characterized in that, The permeation system consists of 24 kg of commercial-grade tea saponin and 4 kg of commercial-grade natural trehalose; the antibacterial and repellent system consists of 16 kg of commercial-grade Sophora flavescens-citronella-peppermint compound plant extract, 5 kg of commercial-grade licorice extract, and 3 kg of commercial-grade Artemisia argyi extract; the enzyme stabilizing system consists of 4 kg of commercial-grade sorbitol, 2 kg of commercial-grade xanthan gum, and 1 kg of commercial-grade natural plant moisturizing factor; the pH adjuster consists of 1.5 kg of commercial-grade citric acid, 0.5 kg of commercial-grade sodium bicarbonate, and 1 kg of commercial-grade natural buffer; the leaf-protecting synergistic ingredient is 1 kg of commercial-grade natural plant amino acids; and the commercial auxiliary ingredients consist of 0.3 kg of food-grade defoamer and 0.2 kg of commercial-grade natural biological preservative.
4. The bio-enzyme insecticide for leafy grasses according to claim 1, characterized in that, The chitinase, neutral protease, amylase, lipase, and polypeptide enzymes mentioned are all commercially available bulk-purchased raw materials; the tea saponin, trehalose, plant extracts, licorice extract, artemisia extract, sorbitol, xanthan gum, natural plant moisturizing factors, natural buffers, natural plant amino acids, defoamers, and natural biological preservatives mentioned are all food-grade or agricultural biological-grade, free of alcohol, chemical preservatives, strong acids and alkalis, and biodegradable.
5. The bio-enzyme insecticide for leafy grasses according to claim 1, characterized in that, The natural buffer can adjust the pH of the insecticide to 5.5-8.0; the natural plant moisturizing factor can enhance the activity and stability of enzymes under high and low temperature environments; the natural biological preservative can extend the shelf life of the finished product to 18 months.
6. A method for preparing a bio-enzyme insecticide specifically for leafy grasses as described in any one of claims 1-5, characterized in that, Includes the following steps: Step 1: Pre-production preparation, sampling and testing of raw materials, cleaning, disinfecting and drying of production equipment, and adjusting the temperature of the production workshop to 15℃-25℃; Step 2: Dechlorination treatment. Treat tap water to dechlorinated purified water, test the pH value to 5.5-7.5, pour it into a mixing tank, and add 70%-85% of the total amount of dechlorinated purified water. Step 3: Enzyme solution preparation. Turn on the mixing tank and stir at medium speed. Add chitinase, neutral protease, amylase, lipase and peptidase in sequence. Stir until dissolved evenly after each addition. Step 4: Add penetrating ingredients, including tea saponin and natural trehalose, along with an antifoaming agent, and stir well; Step 5: Add antibacterial and repellent ingredients. Add the compound plant extracts of Sophora flavescens, lemongrass, and peppermint, licorice extract, and artemisia argyi extract, and stir well. Step 6: Add enzyme-stabilizing and leaf-protecting ingredients, including sorbitol, xanthan gum, natural plant moisturizing factors, and natural plant amino acids, and stir until completely dissolved; Step 7: Precisely adjust the pH by slowly adding citric acid, sodium bicarbonate, and natural buffer while stirring and measuring the pH value to stabilize it between 5.5 and 8.
0. Step 8: Adjust volume and homogenize, add the remaining dechlorinated purified water to the total volume, increase the stirring speed, and homogenize; Step 9: Defoaming and filtration. Let the mixture stand to defoam naturally, then filter it through a precision filtration device. Step 10: Filling and sealing. Fill to commercial specifications and seal immediately. Step 11: Finished product storage. Place the finished products in a cool, dry, and well-ventilated warehouse, with the temperature controlled between 5℃ and 30℃. Arrange them by batch and label them properly.
7. The preparation method according to claim 6, characterized in that, In step 2, the amount of dechlorinated purified water added is 80% of the total volume; in step 3, the stirring speed is 60-80 r / min, and each enzyme is added and stirred for 5 minutes; in step 8, after replenishing the dechlorinated purified water, the speed is adjusted to 80-100 r / min, and homogenized and stirred for 15 minutes; in step 9, it is allowed to stand for 30 minutes to naturally defoam, and then filtered through a 200-mesh precision filter.
8. The preparation method according to claim 6, characterized in that, The method can scale batch sizes proportionally to adapt to production in factories of different sizes.
9. A method for applying a bio-enzyme insecticide specifically for leafy grasses as described in any one of claims 1-5, characterized in that, Pest control for different growth stages of leafy plants, specifically including: (1) Seedling stage prevention: Dilute with water at a ratio of 1:200 to 1:300 and spray on both sides of the leaves once every 5 to 10 days; (2) Prevention and control during the growing season: For routine prevention, dilute with water at a ratio of 1:150 to 1:
250. During the peak period of pests, dilute with water at a ratio of 1:30 to 1:
100. Repeat every 3 to 7 days for 2 to 3 consecutive times. (3) Post-harvest recovery period: Dilute with water at a ratio of 1:150 to 1:250 and spray on new leaves and roots once every 7 to 14 days; (4) Control of underground pests: Dilute with water at a ratio of 1:250 to 1:500 and apply as a root irrigation treatment. Use 600-800g of the original solution per mu after diluting with water.
10. The application method according to claim 9, characterized in that, For seedling prevention, dilute with water at a ratio of 1:250 and apply once every 7 days; for routine prevention during the growing season, dilute with water at a ratio of 1:200; during periods of high pest incidence, dilute with water at a ratio of 1:50 to 1:80 and apply once every 3-5 days; during the recovery period after harvest, dilute with water at a ratio of 1:200 and apply once every 10 days; for control of underground pests, dilute with water at a ratio of 1:300 to 1:
400. The insecticide dilution solution should be prepared and used immediately, and used up within 24 hours; it should not be mixed with strong acid or strong alkali pesticides or fungicides, but can be mixed with organic fertilizers, biological agents, and neutral foliar fertilizers; use should be stopped 3 days before the harvest of leafy grasses.