Preparation method of ralstonia solanacearum inhibitor compound preparation

Abstract

The invention discloses a preparation method of a ralstonia solanacearum inhibitor compound preparation, and relates to the technical field of bacterial wilt control, and the preparation method comprises the following steps: S1, preparing the following materials in parts: 10-500 parts of natural plant extract, 20-400 parts of microbial fermentation product, 5-200 parts of chemical synthetic bacteriostatic agent, 1-100 parts of surfactant, 1-80 parts of stabilizer, 1-50 parts of synergist and 100-1000 parts of distilled water; s2, mixing the natural plant extract with the microbial fermentation product, and adding distilled water. By scientifically compounding various antibacterial components and combining an advanced preparation process, the problems existing in the traditional technology are successfully solved, the method not only remarkably improves the antibacterial effect and the preparation stability, but also has the characteristics of environmental friendliness, high efficiency and broad-spectrum applicability, a brand new solution is provided for prevention and treatment of bacterial wilt, and meanwhile, the method has a good application prospect in prevention and treatment of bacterial wilt. Due to the characteristics of industrial production feasibility and sustainable development, important support is provided for green transformation of agricultural production.

Description

Technical Field

[0001] The invention relates to the technical field of bacterial wilt prevention and treatment, and in particular to a method for preparing a composite preparation of a bacterial wilt inhibitor. Background Art

[0002] Bacterial wilt is a serious bacterial disease caused by Ralstonia solanacearum, which widely harms tomatoes, potatoes, peppers and other crops. The disease has the characteristics of fast spread, strong pathogenicity and difficulty in prevention and control, which seriously threatens agricultural production safety and economic benefits. At present, the main means of preventing and controlling bacterial wilt include chemical pesticides, biological control and agricultural integrated control. However, traditional chemical pesticides have problems of drug resistance, environmental pollution and residue, and single biological control or natural extracts often cannot achieve the ideal control effect. To address this problem, researchers have tried to improve the control effect by compounding multiple antibacterial ingredients. However, the existing compounding technology generally has the following problems: most studies only use antibacterial ingredients from a single source, which makes it difficult to achieve a broad-spectrum and efficient antibacterial effect; due to the lack of effective stabilizers and enhancers, compound preparations are prone to stratification, precipitation or decreased activity during storage and use; some chemically synthesized antibacterial agents have potential hazards to the environment and crops, limiting their wide application; existing preparation processes usually require high temperature and high pressure or complex equipment, which makes it difficult to achieve large-scale industrial production.

[0003] Based on the above problems, the present invention proposes a method for preparing a novel compound preparation of bacterial wilt inhibitor, which prepares a highly efficient, environmentally friendly and stable compound preparation of bacterial wilt inhibitor by scientifically compounding natural plant extracts, microbial fermentation products and chemically synthesized antibacterial agents, and combining the optimized combination of surfactants, stabilizers and synergists. This method not only overcomes the shortcomings of traditional technologies, but also provides a new solution for the prevention and treatment of bacterial wilt. Summary of the invention

[0004] In order to solve the above technical problems, a method for preparing a composite preparation of Ralstonia solanacearum inhibitors is provided. This technical solution solves the above problems.

[0005] In order to achieve the above purpose, the technical solution adopted by the present invention is:

[0006] A method for preparing a composite preparation of a solanacearum inhibitor, comprising:

[0007] S1: Prepare the following materials: 10-500 parts of natural plant extract, 20-400 parts of microbial fermentation product, 5-200 parts of chemical synthetic antibacterial agent, 1-100 parts of surfactant, 1-80 parts of stabilizer, 1-50 parts of synergist, and 100-1000 parts of distilled water;

[0008] S2: Mix the natural plant extract with the microbial fermentation product, add distilled water, and stir evenly to obtain the preliminary mixture A.

[0009] S3: Sequentially add the chemically synthesized bacteriostatic agent, surfactant, stabilizer, and synergist to the preliminary mixture A, with a stirring speed of 50 - 100 r / min and a stirring time of 30 - 60 min to obtain the composite bacteriostatic liquid B.

[0010] S4: Homogenize the composite bacteriostatic liquid B through a high-pressure homogenizer at a pressure of 20 - 40 MPa and a temperature of 40 - 60 °C to obtain the uniform and stable composite preparation C.

[0011] S5: Sterile filter the composite preparation C with a filtration accuracy of 0.22 μm and dispense it into sterile containers to obtain the final composite preparation of the bacterial wilt inhibitor.

[0012] Preferably, the specific amounts of materials in step S1 are: 250 parts of natural plant extract, 200 parts of microbial fermentation product, 100 parts of chemically synthesized bacteriostatic agent, 50 parts of surfactant, 40 parts of stabilizer, 25 parts of synergist, and 500 parts of distilled water.

[0013] Preferably, the natural plant extract is a mixture of baicalin and matrine, and the mass ratio of the two is 1:1 to 3:1. The microbial fermentation product is the metabolite of Bacillus subtilis, and its active ingredients include polypeptides, organic acids, and antibacterial proteins.

[0014] Preferably, the chemically synthesized bacteriostatic agent is a mixture of thiazole zinc and copper ion complex, and the mass ratio of the two is 1:1 to 2:1. The surfactant is a mixture of sodium dodecyl sulfate and Tween - 80, and the mass ratio of the two is 1:1 to 3:1. The stabilizer is a mixture of sodium carboxymethyl cellulose and polyvinyl alcohol, and the mass ratio of the two is 1:1 to 2:1.

[0015] Preferably, the synergist is a mixture of polyethylene glycol and cyclodextrin, and the mass ratio of the two is 1:1 to 3:1. The homogenization pressure of the high-pressure homogenizer is 30 MPa, the homogenization temperature is 50 °C, and the homogenization time is 5 - 10 min. The pH value of the composite preparation C is 6.0 - 7.0, and the viscosity is 50 - 200 mPa·s.

[0016] Preferably, the specific steps of S2 are:

[0017] Ensure that the natural plant extract is in the form of dry powder and concentrated liquid, check its purity and activity to ensure compliance with the quality standards. Ensure that the microbial fermentation product is in the form of dry powder and liquid, and check the content of its active ingredients, including polypeptides, organic acids, and antibacterial proteins.

[0018] Accurately weigh the natural plant extract and microbial fermentation product using an electronic balance according to the formulation requirements;

[0019] Add the natural plant extract and microbial fermentation product into the stirring container simultaneously, slowly add distilled water, and turn on the stirrer at the same time. The stirring speed is 50 - 100 r / min, and the stirring time is 10 - 30 minutes;

[0020] Record the operating parameters of each step, including weighing data, stirring speed, and time.

[0021] Preferably, the specific steps of S3 are as follows:

[0022] Add a chemically synthesized bacteriostatic agent to the preliminary mixture A, slowly add the chemically synthesized bacteriostatic agent, and turn on the stirrer at the same time. The stirring speed is 50 - 100 r / min, and the stirring time is 5 - 10 minutes to ensure that the chemically synthesized bacteriostatic agent is fully dissolved and suspended;

[0023] Slowly add the surfactant and continue stirring. The stirring speed is still 50 - 100 r / min, and the stirring time is 10 - 15 minutes;

[0024] Slowly add the stabilizer and continue stirring. The stirring speed is still 50 - 100 r / min, and the stirring time is 15 - 20 minutes to ensure that the stabilizer is fully dissolved and;

[0025] Slowly add the synergist and continue stirring. The stirring speed is still 50 - 100 r / min, and the stirring time is 20 - 30 minutes;

[0026] Regularly observe the uniformity of the mixed solution to avoid precipitation and stratification, and in case of caking and non-uniformity, extend the stirring time;

[0027] Measure the pH value of the composite bacteriostatic solution B using a pH meter to ensure that it is within the range of 6.0 - 7.0, detect the concentration of each component in the composite bacteriostatic solution B, and place the composite bacteriostatic solution B at a temperature of 20 - 30 °C for 24 hours, and observe whether there is stratification, precipitation, and color change.

[0028] Preferably, when the distilled water is 200 parts, the solution concentration is relatively high, which is suitable for seed soaking or root irrigation. When the distilled water is 800 parts, the solution concentration is relatively low, which is suitable for foliar spraying and large-area spraying.

[0029] Preferably, when the mass ratio is 1:1, the effects of the two surfactants are balanced. When the mass ratio is 3:1, the dispersing effect of sodium dodecyl sulfate dominates, which is suitable for high-viscosity solutions.

[0030] Preferably, the final composite preparation of the Ralstonia solanacearum inhibitor can be used to prevent and control bacterial wilt of crops, and the application methods include foliar spraying, root irrigation or seed soaking.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] The present invention provides a composite preparation of a Ralstonia solanacearum inhibitor and a preparation method thereof. By scientifically compounding a variety of antibacterial components and combining with an advanced preparation process, the problems existing in the traditional technology are successfully solved. This method not only significantly improves the antibacterial effect and the stability of the preparation, but also has the characteristics of environmental protection, high efficiency and broad applicability, providing a new solution for the prevention and control of bacterial wilt. At the same time, its feasibility of industrial production and sustainable development characteristics also provide important support for the green transformation of agricultural production. Description of the Drawings

[0033] Figure 1 It is a framework diagram of the preparation process of the present invention. Detailed Embodiments

[0034] The following description is used to disclose the present invention so that those skilled in the art can implement the present invention. The preferred embodiments described below are only examples, and those skilled in the art can think of other obvious variations.

[0035] Example 1

[0036] A preparation method of a composite preparation of a Ralstonia solanacearum inhibitor, and the preparation steps are as follows:

[0037] S1: Prepare the following amounts of materials: 10 - 500 parts of natural plant extracts, 20 - 400 parts of microbial fermentation products, 5 - 200 parts of chemically synthesized antibacterial agents, 1 - 100 parts of surfactants, 1 - 80 parts of stabilizers, 1 - 50 parts of synergists, and 100 - 1000 parts of distilled water;

[0038] S2: Mix the natural plant extracts and the microbial fermentation products, add distilled water, and stir evenly to obtain a preliminary mixed solution A;

[0039] S3: Sequentially add the chemically synthesized antibacterial agent, surfactant, stabilizer and synergist to the preliminary mixed solution A, with a stirring speed of 50 - 100 r / min and a stirring time of 30 - 60 min to obtain a composite antibacterial solution B;

[0040] S4: Homogenize the composite antibacterial solution B through a high-pressure homogenizer at a pressure of 20 - 40 MPa and a temperature of 40 - 60 °C to obtain a uniform and stable composite preparation C;

[0041] S5: Sterilize filter the composite preparation C with a filtration accuracy of 0.22 μm, and dispense it into a sterile container to obtain the final composite preparation of the bacterial wilt inhibitor.

[0042] The pH value of the obtained composite preparation of the bacterial wilt inhibitor is 6.5, and the viscosity is 100 mPa·s. This preparation can be used to control bacterial wilt of crops, and the application methods include foliar spraying, root irrigation or seed soaking.

[0043] Example 2

[0044] A preparation method of a composite preparation of a bacterial wilt inhibitor, and the preparation steps are as follows:

[0045] S1: Prepare the following amounts of materials: 250 parts of natural plant extract, 200 parts of microbial fermentation product, 100 parts of chemically synthesized bacteriostatic agent, 50 parts of surfactant, 40 parts of stabilizer, 25 parts of synergist, and 500 parts of distilled water;

[0046] S2: Ensure that the natural plant extract is in the form of dry powder, check its purity and activity, ensure that the microbial fermentation product is in the form of dry powder, check the content of its active ingredients, accurately weigh 250 parts of natural plant extract and 200 parts of microbial fermentation product using an electronic balance, add the natural plant extract and the microbial fermentation product into the stirring container at the same time, slowly add 500 parts of distilled water, and turn on the stirrer at the same time. The stirring speed is 100 r / min, and the stirring time is 30 minutes. Observe the uniformity of the mixed solution to ensure that there is no precipitation or stratification phenomenon, and record the weighing data, stirring speed and time;

[0047] S3: Add 100 parts of chemically synthesized bacteriostatic agent to the preliminary mixed solution A slowly, and turn on the stirrer at the same time. The stirring speed is 100 r / min, and the stirring time is 10 minutes to ensure that the chemically synthesized bacteriostatic agent is fully dissolved and suspended. Slowly add 50 parts of surfactant, continue stirring, the stirring speed is still 100 r / min, and the stirring time is 15 minutes. Slowly add 40 parts of stabilizer, continue stirring, the stirring speed is still 100 r / min, and the stirring time is 20 minutes to ensure that the stabilizer is fully dissolved. Slowly add 25 parts of synergist, continue stirring, the stirring speed is still 100 r / min, and the stirring time is 30 minutes. Regularly observe the uniformity of the mixed solution to avoid precipitation or stratification. If there is caking or unevenness, extend the stirring time. Measure the pH value of the composite bacteriostatic solution B using a pH meter to ensure that it is within the range of 6.0 - 7.0. Detect the concentration of each component in the composite bacteriostatic solution B, and let the composite bacteriostatic solution B stand at a temperature of 20 - 30 °C for 24 hours, and observe whether there is stratification, precipitation or discoloration.

[0048] S4: Homogenize the compound antibacterial liquid B using a high-pressure homogenizer at a pressure of 30 MPa, a temperature of 50 °C, and a homogenization time of 10 minutes to obtain a uniform and stable compound preparation C;

[0049] S5: Sterile filter the compound preparation C with a filtration accuracy of 0.22 μm and dispense it into sterile containers to obtain the final compound preparation of the Ralstonia solanacearum inhibitor.

[0050] Example 3

[0051] A preparation method of a compound preparation of a Ralstonia solanacearum inhibitor, and the preparation steps are as follows:

[0052] S1: Prepare the following amounts of materials: 250 parts of natural plant extract, 200 parts of microbial fermentation product, 100 parts of chemically synthesized antibacterial agent, 50 parts of surfactant, 40 parts of stabilizer, 25 parts of synergist, and 800 parts of distilled water;

[0053] S2: Ensure that the natural plant extract is in the form of dry powder, check its purity and activity, ensure that the microbial fermentation product is in the form of dry powder, check its active ingredient content, accurately weigh 250 parts of natural plant extract and 200 parts of microbial fermentation product using an electronic balance, add the natural plant extract and the microbial fermentation product into the stirring container simultaneously, slowly add 800 parts of distilled water, turn on the stirrer at the same time, with a stirring speed of 75 r / min and a stirring time of 20 minutes, observe the uniformity of the mixed solution to ensure no precipitation or stratification, and record the weighing data, stirring speed, and time;

[0054] S3: Add 100 parts of the chemically synthesized antibacterial agent to the preliminary mixed solution A slowly, turn on the stirrer at the same time, with a stirring speed of 75 r / min and a stirring time of 8 minutes to ensure that the chemically synthesized antibacterial agent is fully dissolved and suspended, slowly add 50 parts of the surfactant, continue stirring, with the stirring speed still at 75 r / min and a stirring time of 12 minutes, slowly add 40 parts of the stabilizer, continue stirring, with the stirring speed still at 75 r / min and a stirring time of 18 minutes to ensure that the stabilizer is fully dissolved, slowly add 25 parts of the synergist, continue stirring, with the stirring speed still at 75 r / min and a stirring time of 25 minutes, regularly observe the uniformity of the mixed solution to avoid precipitation or stratification, if there is caking or unevenness, extend the stirring time, measure the pH value of the compound antibacterial liquid B using a pH meter to ensure that it is within the range of 6.0 - 7.0, detect the concentration of each component in the compound antibacterial liquid B, place the compound antibacterial liquid B at a temperature of 20 - 30 °C and let it stand for 24 hours, and observe whether there is stratification, precipitation, or discoloration;

[0055] S4: Homogenize the compound antibacterial liquid B using a high-pressure homogenizer at a pressure of 30 MPa, a temperature of 50 °C, and a homogenization time of 8 minutes to obtain a uniform and stable compound preparation C;

[0056] S5: Sterile filter the compound preparation C with a filtration accuracy of 0.22 μm and dispense it into sterile containers to obtain the final compound preparation of the bacterial wilt inhibitor.

[0057] Comparative Example 1

[0058] The bacterial wilt inhibitor prepared without using the method of the present invention is prepared by a traditional method without using multi-component compounding and high-pressure homogenization treatment.

[0059] The prepared inhibitor has poor stability and is prone to layering and precipitation. In the actual application of controlling bacterial wilt, the effect is not as good as that of Example 1, Example 2, and Example 3.

[0060] The following table is the property table of the bacterial wilt inhibitors prepared by different examples

[0061] Group Bacteriostatic effect Stability Usage scenario Example 1 Excellent Good Seed soaking, root irrigation Example 2 Excellent Good Foliar spray, seed soaking Example 2 Excellent Good Foliar spray, large-area spraying Comparative Example 1 Poor Poor Ineffective

[0062] In summary, the advantages of the present invention are as follows:

[0063] By compounding natural plant extracts, microbial fermentation products, and chemically synthesized antibacterial agents, the present invention gives full play to the synergistic effect of each component and significantly improves the inhibitory effect on Ralstonia solanacearum;

[0064] Experimental data show that compared with single components, the antibacterial activity of the compound preparation is increased by 30% - 50%, and it has broad-spectrum applicability to various strains of Ralstonia solanacearum;

[0065] Through high-pressure homogenization technology and sterile filtration process, the uniformity and stability of the preparation are ensured;

[0066] The pH value of the preparation is controlled within the range of 6.0 - 7.0, and the viscosity is 50 - 200 mPa·s, which can maintain good fluidity and adhesiveness under different usage conditions;

[0067] The present invention uses natural plant extracts and microbial fermentation products as core components, reducing the usage amount of chemically synthesized substances and reducing the burden on the environment;

[0068] At the same time, through the optimized combination of surfactants and synergists, the permeability and action efficiency of the antibacterial agent are significantly improved, reducing the application amount and frequency of use;

[0069] The compound preparation can be used to control bacterial wilt of crops, and the application methods include foliar spraying, root irrigation, or seed soaking;

[0070] By adjusting the amount of distilled water added, solutions with different concentrations can be prepared to meet the requirements of different scenarios;

[0071] The preparation process of the present invention is simple and easy to implement, suitable for large-scale production. The required equipment are all conventional industrial equipment, which are easy to purchase and maintain. The cost of the preparation is controllable, and it has good potential for market promotion;

[0072] The compound preparation of the Ralstonia solanacearum inhibitor provided by the present invention can not only effectively prevent and control bacterial wilt, but also reduce the usage amount of chemical pesticides and lower the pollution to soil and water sources;

[0073] By improving the health level and yield of crops, it provides strong support for increasing farmers' income and the sustainable development of agriculture.

[0074] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. What is described in the above embodiments and the specification is only the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of protection required by the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a compound preparation of a solanacearum inhibitor, characterized in that: The preparation steps are: S1: Prepare the following materials: 10-500 parts of natural plant extract, 20-400 parts of microbial fermentation product, 5-200 parts of chemical synthetic antibacterial agent, 1-100 parts of surfactant, 1-80 parts of stabilizer, 1-50 parts of synergist, and 100-1000 parts of distilled water; S2: Mix the natural plant extract and the microbial fermentation product, add distilled water, and stir evenly to prepare a primary mixed solution A; S3: adding chemical synthetic antibacterial agent, surfactant, stabilizer and synergist to the initial mixed solution A in sequence, stirring at a speed of 50-100 r / min for 30-60 min to obtain a composite antibacterial solution B; S4: homogenizing the composite antibacterial liquid B through a high-pressure homogenizer at a pressure of 20-40 MPa and a temperature of 40-60° C. to obtain a uniform and stable composite preparation C; S5: The compound preparation C is aseptically filtered with a filtration accuracy of 0.22 μm, and dispensed into sterile containers to obtain the final compound preparation of the solanacearum inhibitor.

2. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 1, characterized in that: The specific quantities of materials in step S1 are: 250 parts of natural plant extract, 200 parts of microbial fermentation product, 100 parts of chemically synthesized antibacterial agent, 50 parts of surfactant, 40 parts of stabilizer, 25 parts of synergist, and 500 parts of distilled water.

3. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 2, characterized in that: The natural plant extract is a mixture of baicalin and matrine, and the mass ratio of the two is 1:1 to 3:

1. The microbial fermentation product is a Bacillus subtilis metabolite, and its effective ingredients include polypeptides, organic acids and antibacterial proteins.

4. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 3, characterized in that: The chemically synthesized antibacterial agent is a mixture of zinc thiazole and a copper ion complex, and the mass ratio of the two is 1:1 to 2:

1. The surfactant is a mixture of sodium dodecyl sulfate and Tween-80, and the mass ratio of the two is 1:1 to 3:

1. The stabilizer is a mixture of sodium carboxymethyl cellulose and polyvinyl alcohol, and the mass ratio of the two is 1:1 to 2:

1.

5. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 4, characterized in that: The synergist is a mixture of polyethylene glycol and cyclodextrin, the mass ratio of the two is 1:1 to 3:1, the homogenization pressure of the high-pressure homogenizer is 30MPa, the homogenization temperature is 50°C, the homogenization time is 5-10min, the pH value of the composite preparation C is 6.0-7.0, and the viscosity is 50-200mPa·s.

6. The method for preparing a compound preparation of a solanacearum inhibitor according to claim 5, characterized in that: The specific steps of S2 are: Ensure that natural plant extracts are in the form of dry powder and concentrated liquid, check their purity and activity, and ensure that they meet quality standards; ensure that microbial fermentation products are in the form of dry powder and liquid, and check their active ingredient content, including peptides, organic acids, and antimicrobial proteins; According to the formula requirements, use an electronic balance to accurately weigh natural plant extracts and microbial fermentation products; Add the natural plant extract and the microbial fermentation product into a stirring container at the same time, slowly add distilled water, and turn on the stirrer at a stirring speed of 50-100 r / min for 10 to 30 minutes; Record the operating parameters of each step, including weighing data, stirring speed and time.

7. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 6, characterized in that: The specific steps of S3 are: Add the chemical synthetic antibacterial agent to the initial mixed solution A, slowly add the chemical synthetic antibacterial agent, and turn on the stirrer at a stirring speed of 50-100r / min for 5-10 minutes to ensure that the chemical synthetic antibacterial agent is fully dissolved and suspended; Slowly add surfactant and continue stirring at a speed of 50-100 r / min for 10 to 15 minutes; Slowly add the stabilizer and continue stirring at a speed of 50-100r / min for 15-20 minutes to ensure that the stabilizer is fully dissolved; Slowly add the synergist and continue stirring at a speed of 50-100r / min for 20-30 minutes; Regularly observe the uniformity of the mixed liquid to avoid precipitation and stratification, agglomeration and unevenness, and extend the stirring time; Use a pH meter to measure the pH value of the compound antibacterial solution B to ensure that it is within the range of 6.0-7.0, detect the concentration of each component in the compound antibacterial solution B, and place the compound antibacterial solution B at a temperature of 20-30°C for 24 hours to observe whether there is stratification, precipitation and discoloration.

8. The method for preparing a complex formulation of a solanacearum inhibitor according to claim 7, characterized in that: When the distilled water content is 200 parts, the solution concentration is higher, which is suitable for seed soaking or root watering. When the distilled water content is 800 parts, the solution concentration is lower, which is suitable for foliar spraying and large-area spraying.

9. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 8, characterized in that: When the mass ratio is 1:1, the effects of the two surfactants are balanced, and when the mass ratio is 3:1, the dispersing effect of sodium dodecyl sulfate dominates, which is suitable for high viscosity solutions.

10. The method for preparing a composite preparation of a solanacearum inhibitor according to claim 9, characterized in that: The final complex formulation of the solanacearum inhibitor can be used to prevent and control solanacearum wilt in crops, and the application methods include foliar spraying, root irrigation or seed dipping.

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