Toosendanin microbial biopesticide as well as preparation method and application thereof

Abstract

The invention relates to a toosendanin microbial biopesticide, which is prepared from the following raw materials in parts by weight: 70 to 90 parts of microbial fermentation liquor, 200 to 250 parts of toosendanin solution and 250 to 300 parts of mineral-derived potassium fulvate carrier, the surface of mineral-derived potassium fulvate is coated with a chitosan protective film, the microbial fermentation liquor comprises bacillus thuringiensis, and the chitosan protective film is coated with a chitosan protective film. The toosendanin solution is prepared from 40 to 60 weight percent of toosendanin. The application has the effects of prolonging the storage period of the microbial biopesticide and improving the stability of the control effect.

Description

Technical Field

[0001] The present application relates to the field of biological pesticides, and in particular to toosendanin microbial biological pesticides and preparation methods and applications thereof. Background Art

[0002] In agricultural production, pesticides used to control pests and diseases mainly include chemical pesticides and biological pesticides. Among them, biological pesticides have attracted much attention because of their safety, effectiveness and pollution-free characteristics. The use of biological pesticides to control pests and diseases has developed into one of the important research topics of plant protection scientists at home and abroad.

[0003] Biopesticides are classified according to their ingredients and sources, including plant-based pesticides, animal-based pesticides and microbial pesticides. Among them, microbial pesticides have the advantages of being easily degradable in the natural environment and harmless, and have become one of the important components of biopesticides.

[0004] Regarding the above-mentioned related technologies, since microbial pesticides are easily affected by the external environment during storage, the storage period is short, resulting in unstable control effects, which limits the promotion and use of biological pesticides. Summary of the invention

[0005] In order to improve the storage period and stability of the control effect of microbial pesticides, the present application provides a toosendan microbial pesticide and a preparation method thereof.

[0006] In the first aspect, the present application provides a Toosendan microbial biological pesticide, which adopts the following technical solution:

[0007] A toosendan microbial biological pesticide comprises the following raw materials in parts by weight: 70-90 parts of microbial fermentation liquid, 200-250 parts of toosendan solution and 250-300 parts of mineral source potassium humate carrier, wherein the surface of the mineral source potassium humate is coated with a chitosan protective film, the microbial fermentation liquid comprises Bacillus thuringiensis, and the toosendan solution comprises 40-60wt% of toosendan.

[0008] By adopting the above technical scheme, the microbial fermentation liquid and the toosendan solution are used in combination to synergistically play an insecticidal role. Mineral potassium humate serves as a carrier of active ingredients such as the microbial fermentation liquid and the toosendan solution, which helps to reduce the loss of active ingredients of the biological pesticide due to the influence of the external environment during storage. In addition, the chitosan protective film wrapped on the surface of the mineral potassium humate further improves the storage period of the biological pesticide and the stability of the control effect.

[0009] Bacillus thuringiensis can secrete toxin proteins, which in turn affect the growth and development of pests, and has a good preventive and control effect on pests that harm the development and production of crops; toosendanin is an active ingredient extracted from the root bark of the toosendan tree. When pests come into contact with toosendanin, it can block the conduction of nerve centers, destroy intestinal tissue and various respiratory and metabolic functions, affect digestion and absorption, and cause pests to refuse to eat, resulting in abnormal growth and development and death.

[0010] Mineral potassium humate has a unique cavity structure, and has a strong adsorption capacity for toosendan solution and microbial fermentation liquid, and can release them in a controlled manner. In addition, due to the strong hydrophilicity of mineral potassium humate, it is more helpful for biological pesticides to penetrate into the body of pests when used, thereby achieving the effect of preventing and controlling pests.

[0011] Optionally, it comprises the following raw materials in parts by weight: 75-85 parts of microbial fermentation broth, 220-240 parts of toosendan solution and 260-290 parts of mineral source potassium humate.

[0012] By adopting the above technical scheme, when the present application selects 75-85 parts of microbial fermentation liquid, 220-240 parts of toosendan solution and 260-290 parts of mineral potassium humate, the storage period and stability of the control effect of the biological pesticide prepared therefrom have no obvious substantial difference, and when 80 parts of microbial fermentation liquid, 230 parts of toosendan solution and 275 parts of mineral potassium humate are used, the biological pesticide prepared has the best effect.

[0013] Optionally, the surface of the mineral source potassium humate is coated with a chitosan protective film, and the preparation method thereof is as follows:

[0014] Chitosan is mixed with 1wt% acetic acid solution, and then mineral potassium humate is added, wherein the ratio of mineral potassium humate to chitosan is 1:(0.8-1.2), and ultrasonic treatment is performed in a water bath for 0.8-1.2h, and the mixture is dried after cooling to obtain mineral potassium humate wrapped with a chitosan protective film.

[0015] By adopting the above technical scheme, chitosan is connected by intramolecular and intermolecular hydrogen bonds to form a film-forming solution, and after drying, a protective film is formed and wrapped on the surface of the mineral-derived potassium humate. This is because the amino and hydroxyl groups on the chitosan molecules interact with the hydroxyl groups on the surface of the mineral-derived potassium humate through hydrogen bonds to form a tight bond, which protects the active ingredients adsorbed in the mineral-derived potassium humate and prolongs the storage period of the biological pesticide and the stability of the control effect.

[0016] Optionally, the surface of the mineral source potassium humate is also embedded with methionine iodine particles, and the preparation method thereof is as follows:

[0017] Chitosan and methionine iodine particles are mixed with 1wt% acetic acid solution, and then mineral potassium humate is added, wherein the amount ratio of mineral potassium humate to chitosan and methionine iodine particles is 1:(0.8-1.2):(0.2-0.4), and ultrasonic treatment is performed in a water bath for 0.8-1.2h, and the mixture is cooled and dried to obtain mineral potassium humate with methionine iodine particles embedded on the surface.

[0018] By adopting the above technical solution, since the methionine iodine particles are volatile, when the biological pesticide is spread in the field, the methionine iodine particles come into contact with the air and are affected by the humidity, the volume of the methionine iodine particles gradually decreases, making the chitosan protective film less dense, and having many tiny micropores on its surface, which facilitates the continuous diffusion of the active ingredients in the mineral potassium humate carrier, so that the biological pesticide can kill pests efficiently and durably.

[0019] Optionally, the microbial fermentation broth further comprises the following raw materials in percentage by weight: 40-60 wt % of Bacillus thuringiensis, 20-30 wt % of Bacillus brevis and 20-30 wt % of Bacillus sphaericus.

[0020] By adopting the above technical scheme, Bacillus thuringiensis, Bacillus brevis and Bacillus sphaericus can all secrete toxin proteins, thereby affecting the growth and development of pests, further improving the effect of pest control.

[0021] Optionally, the toosendan solution comprises raw materials in the following percentages: 40-60 wt % toosendan, 10-20 wt % glycine and 10-20 wt % methylpropanediol.

[0022] By adopting the above technical solution, since Toosendan itself is insoluble in water, Toosendan solution obtained by compounding Toosendan, methylpropanediol and glycine can promote better dispersion of Toosendan, so as to better adsorb on the mineral potassium humate carrier. Glycine is an amphoteric organic compound. The addition of glycine reduces the use of methylpropanediol and promotes the dispersion of Toosendan. Glycine can provide organic nitrogen to the soil, provide nutrition to crops, and promote crop growth.

[0023] In the second aspect, the present application provides a method for preparing toosendan microbial biological pesticide, using the following technical scheme:

[0024] A method for preparing toosendan microbial biological pesticide comprises the following steps:

[0025] Under a sterile environment, the microbial fermentation liquid is mixed with the mineral source potassium humate carrier, and then the toosendanin solution is added, mixed, dried at room temperature, and set aside;

[0026] Then the surface of the mineral potassium humate carrier is coated with a chitosan protective film, and the biological pesticide powder is obtained after drying.

[0027] By adopting the above technical scheme, mineral potassium humate is used as a carrier to protect the active ingredients in the microbial fermentation liquid, and then toosendan solution is added to continue adsorption, so that the active ingredients that kill pests can be preserved for a longer time. In addition, the chitosan protective film can ensure its protective effect on the active ingredients. At the same time, during the use of biological pesticides, since the chitosan protective film is not dense, it is beneficial for biological pesticides to kill pests more persistently and ensure the prevention and control effect.

[0028] In a third aspect, the present application provides an application of toosendanin microbial biological pesticide.

[0029] In summary, the present application includes at least one of the following beneficial technical effects:

[0030] 1. The present application uses a compound of microbial fermentation broth and toosendan solution to synergistically kill insects. Mineral potassium humate is used as a carrier of active ingredients such as microbial fermentation broth and toosendan solution, which helps to reduce the loss of active ingredients caused by the external environment during the storage of biological pesticides. The chitosan protective film wrapped on the surface of mineral potassium humate further improves the storage period of the biological pesticide and the stability of the control effect.

[0031] 2. When the present application uses 75-85 parts of microbial fermentation liquid, 220-240 parts of Toosendan solution and 260-290 parts of mineral-derived potassium humate, the storage period and stability of the control effect of the biological pesticide prepared therefrom have no significant substantial difference, and when 80 parts of microbial fermentation liquid, 230 parts of Toosendan solution and 275 parts of mineral-derived potassium humate are used, the biological pesticide prepared has the best effect;

[0032] 3. The present application uses a chitosan protective film made by adding methionine iodine particles to the surface of the carrier. When the biological pesticide is spread in the field, the methionine iodine particles come into contact with the air and are affected by humidity. The volume of the methionine iodine particles gradually decreases, making the chitosan protective film less dense. There are many tiny micropores on its surface, which facilitates the continuous diffusion of the active ingredients in the mineral potassium humate carrier, so that the biological pesticide can kill pests efficiently and durably. DETAILED DESCRIPTION

[0033] The present application is further described in detail below in conjunction with specific embodiments and comparative examples.

[0034] The following raw materials in this application are all commercially available products, and are intended to fully disclose the raw materials of this application, and should not be construed as limiting the sources of the raw materials.

[0035] Bacillus thuringiensis, bacterial content of 10 billion CFU / g; Bacillus brevis, bacterial content of 10 billion CFU / g; Bacillus sphaericus, bacterial content of 10 billion CFU / g; Toosendanin CAS No.: 58812-37-6, particle size of 120 mesh; mineral potassium fulvic acid carrier, particle size of 60 mesh, humic acid ≥65%, fulvic acid ≥50%, potassium oxide ≥12%, pH: 8-10, purchased from Jinan Qinghai Chemical Co., Ltd.; methionine iodine granules with a molecular formula of C5H 10 INO2S, molecular weight is 275.10787, particle size is 80 mesh; chitosan CAS number: 9012-76-4; glycine CAS number: 56-40-6.

[0036] Example

[0037] Examples 1.1-1.5

[0038] A Toosendan microbial biological pesticide, comprising the following raw materials by weight:

[0039] Microbial fermentation liquid 70, 75, 80, 85, 90 kg, Toosendan solution 200, 220, 230, 240, 250 kg and mineral source potassium humate carrier 250, 260, 275, 290, 300 kg;

[0040] The surface of the mineral-source potassium humate is coated with a chitosan protective film, the microbial fermentation liquid comprises bacillus thuringiensis, and the toosendan solution comprises 40-60wt% toosendan.

[0041] It comprises the following preparation steps:

[0042] Under a sterile environment, the microbial fermentation liquid (Bacillus thuringiensis) and the mineral source potassium humate carrier are mixed, and then the toosendan solution (the content of toosendan is 50wt%) is added, mixed, dried at room temperature, and set aside;

[0043] Then, the surface of the mineral source potassium humate carrier is coated with a chitosan protective film, and the preparation steps are as follows:

[0044] Chitosan was mixed with 1 wt% acetic acid solution, and then the mineral potassium humate prepared above was added. The amount of mineral potassium humate and chitosan was as shown in Table 1. The mixture was ultrasonically treated in a water bath for 1 hour, cooled and dried for 5 hours to obtain mineral potassium humate wrapped with a chitosan protective film, thereby preparing a biological pesticide powder.

[0045] As shown in Table 1, the difference between Examples 1.1-1.5 is that the proportions of the raw materials used are different.

[0046] Table 1 Raw material dosage of Examples 1.1-1.5 (unit: kg)

[0047]

[0048] Examples 2.1-2.3

[0049] A Toosendan microbial biological pesticide, which is different from Example 1.3 in that methionine iodine particles are embedded on the surface of the mineral source potassium humate, and the preparation method thereof is as follows:

[0050] Chitosan, methionine iodine particles and 1wt% acetic acid solution were mixed to a solid-liquid ratio of 2:50, and then potassium humate from mineral sources was added. The dosage ratio of potassium humate from mineral sources to chitosan and methionine iodine particles is shown in Table 2. The mixture was ultrasonically treated in a water bath for 1h, cooled and dried to obtain potassium humate from mineral sources with methionine iodine particles embedded in the chitosan protective film.

[0051] The specific dosage is shown in Table 2

[0052] Table 2 Amount of each raw material used in Examples 2.1-2.3 (unit: kg)

[0053] Example 2.1 Example 2.2 Example 2.3 Mineral Potassium Fulvic Acid 275 275 275 Chitosan 275 275 275 Methionine Iodine Granules 55 82.5 110

[0054] Examples 3.1-3.3

[0055] A toosendan microbial biological pesticide, which is different from Example 2.2 in that Bacillus thuringiensis, Bacillus brevis and Bacillus sphaericus are added to the microbial fermentation broth, and the other raw materials are exactly the same as those in Example 2.2.

[0056] The specific dosage is shown in Table 3.

[0057] Table 3 Amount of each raw material in microbial fermentation broth (unit: kg)

[0058] Example 3.1 Example 3.2 Example 3.3 Bacillus thuringiensis 40 50 60 Bacillus brevis 30 25 20 Bacillus sphaericus 30 25 20

[0059] Examples 4.1-4.3

[0060] A toosendan microbial biological pesticide, which is different from Example 3.2 in that toosendan, glycine and methylpropanediol are added to the toosendan solution, and the other raw materials are the same as those in Example 3.2.

[0061] The specific dosage is shown in Table 4.

[0062] Table 4 The amount of each raw material in the Toosendan solution (unit: kg)

[0063] Example 4.1 Example 4.2 Example 4.3 Toosendanin 40 50 60 Glycine 10 15 20 Methylpropanediol 10 15 20

[0064] Example 5

[0065] A toosendan microbial biopesticide, which is different from Example 4.2 in that glycine is replaced by an equal amount of leucine.

[0066] Comparative Example

[0067] Comparative Example 1

[0068] The preparation method of the biological pesticide in Comparative Example 1 is exactly the same as that in Example 1.3, except that the surface of the mineral potassium humate is not coated with a chitosan protective film.

[0069] Comparative Example 2

[0070] The preparation method of the biological pesticide in Comparative Example 2 is exactly the same as that in Example 1.3, except that the amount of the mineral source potassium humate carrier and chitosan used is 0 kg.

[0071] Comparative Example 3

[0072] The preparation method of the biological pesticide in Comparative Example 2 is exactly the same as that in Example 1.3, except that the amount of toosendan solution used is 0 kg.

[0073] Application Examples

[0074] Application Examples 1.1-1.5

[0075] An application of toosendan microbial biopesticide, the biopesticides prepared in Examples 1.1-1.5 were stored in an environment of 30°C for 1 month, 3 months and 6 months respectively, then mixed with water and spread on the soil around the roots of green vegetables and on the leaves for field experiments, the application rate of the biopesticide was 1kg / mu.

[0076] Application Examples 2.1-2.3

[0077] An application of toosendan microbial biological pesticide, wherein the biological pesticide used is prepared by Examples 2.1-2.3.

[0078] Application Examples 3.1-3.3

[0079] An application of toosendan microbial biological pesticide, wherein the biological pesticide used is prepared by Examples 3.1-3.3.

[0080] Application Examples 4.1-4.3

[0081] An application of toosendan microbial biological pesticide, wherein the biological pesticide used is prepared by Examples 4.1-4.3.

[0082] Application Example 5

[0083] An application of toosendan microbial biological pesticide, wherein the biological pesticide used is prepared by Example 5.

[0084] Comparative Application Examples 1-3

[0085] An application of toosendan microbial biological pesticide, wherein the biological pesticide is prepared by comparative examples 1-3.

[0086] Performance testing

[0087] 1. The field experiment adopts a divided area group test design, with each area of ​​1m 2 The application period is when the vegetables are in the 6-7 leaf stage, and the application is once every 10 days. The efficacy is investigated 10 days and 30 days after the application.

[0088] The biological pesticides corresponding to Example 1.1-1.5, Application Example 2.1-2.3, Application Example 3.1-3.3, Application Example 4.1-4.3, and Application Example 5 were stored in an environment of 30°C for 1 month, 3 months, and 6 months respectively before field experiments. The test results are shown in Table 5.

[0089] 2. The yield increase rate test was carried out according to the corresponding examples 4.1-4.3, application example 5 and application example 3.3. The test results are shown in Table 6.

[0090] Table 5 Field test results of biopesticide application after storage for 1 month, 3 months and 6 months

[0091]

[0092]

[0093] Table 6 Test results of yield increase of biological pesticides in application examples 4.1-4.3, application example 5 and application example 3.3

[0094] Yield increase Application Example 3.2 35.80％ Application Example 4.1 40.60％ Application Example 4.2 41.30％ Application Example 4.3 40.80％ Application Example 5 38.50％

[0095] Combining Application Examples 1.1-1.5 and Comparative Application Examples 1-3 and Table 5, it can be seen that as the storage time of the biological pesticide becomes longer, the pest control effect of Application Examples 1.1-1.5 is significantly better than that of Comparative Application Examples 1-3, and the pest control is durable. When the dosage of each raw material of the biological pesticide adopts the ratio of Application Example 1.3, the effect is better. The analysis reason may be that the microbial fermentation broth and the toosendan solution are used in combination to synergistically play an insecticidal role. Mineral potassium humate serves as a carrier of active ingredients such as microbial fermentation broth and toosendan solution, which helps to reduce the loss of active ingredients in the biological pesticide caused by the influence of the external environment during storage. In addition, the chitosan protective film wrapped on the surface of the mineral potassium humate further improves the storage period of the biological pesticide and the stability of the control effect.

[0096] Bacillus thuringiensis can secrete toxin proteins, which in turn affect the growth and development of pests, and has a good preventive and control effect on pests that harm the development and production of crops; toosendanin is an active ingredient extracted from the root bark of the toosendan tree. When pests come into contact with toosendanin, it can block the conduction of nerve centers, destroy intestinal tissue and various respiratory and metabolic functions, affect digestion and absorption, and cause pests to refuse to eat, resulting in abnormal growth and development and death.

[0097] Mineral potassium humate has a unique cavity structure, and has a strong adsorption capacity for toosendan solution and microbial fermentation liquid, and can release them in a controlled manner. In addition, due to its strong hydrophilicity, it is more conducive to the penetration of biological pesticides into the body of pests when used, thus achieving the effect of pest control.

[0098] Chitosan forms a film-forming solution through intramolecular and intermolecular hydrogen bonds. After drying, it forms a protective film and wraps around the surface of mineral potassium humate. This is because the amino and hydroxyl groups on the chitosan molecules interact with the hydroxyl groups on the surface of mineral potassium humate through hydrogen bonds to form a tight bond, which protects the active ingredients adsorbed in the mineral potassium humate and prolongs the storage period of the biological pesticide and the stability of the control effect.

[0099] Combining Application Examples 2.1-2.3 and Application Example 1.3 and Table 5, it can be seen that the biological pesticides in Application Examples 2.1-2.3 have better control effects. The reason may be that the methionine iodine particles are volatile. When the biological pesticide is spread in the field, the methionine iodine particles come into contact with the air and are affected by humidity. The volume of the methionine iodine particles gradually decreases, making the chitosan protective film less dense. There are many tiny micropores on its surface, which facilitates the continuous diffusion of the active ingredients in the mineral potassium humate carrier, so that the biological pesticide can kill pests efficiently and persistently.

[0100] Combining Application Examples 3.1-3.3 and Application Example 2.2 with Table 5, it can be seen that when the microbial fermentation broth uses Bacillus thuringiensis, Bacillus brevis and Bacillus sphaericus, as the number of microbial species increases, it does not have an adverse effect on the storage stability and control effect of the biological pesticide, and it still has a better control effect after long-term storage.

[0101] Combining Application Examples 4.1-4.3 with Application Examples 3.2 and 5 and Tables 5-6, it can be seen that the biological pesticides in Application Examples 4.1-4.3 are better than those in Application Examples 3.2 and 5. The reason may be that the Toosendan solution obtained by compounding Toosendan, methylpropanediol and glycine can promote better dispersion of Toosendan, so that it can be better adsorbed on the mineral potassium humate carrier. Glycine is an amphoteric organic compound. The addition of glycine reduces the use of methylpropanediol and promotes the dispersion of Toosendan. Glycine can provide organic nitrogen to the soil, provide nutrition to crops, and promote crop growth.

[0102] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereto. Therefore, any equivalent changes made according to the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. A Toosendan microbial biological pesticide, characterized in that: The invention comprises the following raw materials in parts by weight: 70-90 parts of microbial fermentation liquid, 200-250 parts of toosendan solution and 250-300 parts of mineral source potassium humate carrier, the surface of the mineral source potassium humate is coated with a chitosan protective film, the microbial fermentation liquid comprises Bacillus thuringiensis, and the toosendan solution comprises 40-60wt% of toosendan.

2. The toosendan microbial biological pesticide according to claim 1, characterized in that: The method comprises the following raw materials in parts by weight: 75-85 parts of microbial fermentation liquid, 220-240 parts of toosendan solution and 260-290 parts of mineral source potassium fulvic acid.

3. The toosendan microbial biological pesticide according to claim 1, characterized in that: The surface of the mineral source potassium humate is coated with a chitosan protective film, and the preparation method thereof is as follows: Chitosan and 1wt% acetic acid solution are mixed, and then mineral potassium humate is added, wherein the amount ratio of mineral potassium humate to chitosan is 1:(0.8-1.2), and the mixture is ultrasonically treated in a water bath for 0.8-1.2h, cooled and dried to obtain mineral potassium humate wrapped with a chitosan protective film.

4. The toosendan microbial biological pesticide according to claim 3, characterized in that: The surface of the mineral source potassium humate is also embedded with methionine iodine particles, and the preparation method thereof is as follows: Chitosan and methionine iodine particles are mixed with 1wt% acetic acid solution, and then mineral potassium humate is added, wherein the usage ratio of mineral potassium humate to chitosan and methionine iodine particles is 1:(0.8-1.2):(0.2-0.4), and the mixture is ultrasonically treated in a water bath for 0.8-1.2h, cooled and dried to obtain mineral potassium humate embedded with methionine iodine particles.

5. The toosendan microbial biological pesticide according to claim 1, characterized in that: The microbial fermentation liquid also includes the following raw materials in percentage by weight: 40-60 wt% of Bacillus thuringiensis, 20-30 wt% of Bacillus brevis and 20-30 wt% of Bacillus sphaericus.

6. The toosendan microbial biological pesticide according to claim 1, characterized in that: The toosendan solution comprises the following raw materials in percentage: 40-60 wt % toosendan, 10-20 wt % glycine and 10-20 wt % methyl propanediol.

7. A method for preparing the Toosendanin microbial biological pesticide according to any one of claims 1 to 6, characterized in that: The following steps are involved: Under a sterile environment, the microbial fermentation liquid is mixed with the mineral source potassium humate carrier, and then the toosendanin solution is added, mixed, dried at room temperature, and set aside; Then the surface of the mineral source potassium humate carrier is coated with a chitosan protective film, and the biological pesticide powder is obtained after drying.

8. Use of the Toosendan microbial biological pesticide according to any one of claims 1 to 6 in field pest control.

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