阅读说明：本技术 一种颗粒剂、其制备方法及其应用 (Granules, preparation method and application thereof ) 是由 张�杰 杨代斌 耿丽丽 束长龙 袁会珠 王泽宇 于 2021-09-02 设计创作，主要内容包括：本发明涉及一种颗粒剂、其制备方法及其应用。颗粒剂为粒径0.2至2mm的球状物,包括活性组分、第一组分、第二组分、第三组分和第四组分；活性组分为苏云金芽胞杆菌；第一组分为聚乙烯醇、羧甲基纤维素钠、甲基纤维素、黄原胶、聚丙烯酸钠、可溶性淀粉和明胶中的至少一种；第二组分为乙基纤维素、羟乙基纤维素、羟丙基纤维素、醋酸纤维素、预胶化淀粉、交联聚乙烯吡咯烷酮和交联聚丙烯酸钠中的至少一种；第三组分为十二烷基苯磺酸钠、烷基萘磺酸盐甲醛缩合物、木质素磺酸钠和月桂醇聚氧乙烯基醚硫酸钠等中的至少一种；第四组分为膨润土、高岭土、凹凸棒土、硅藻土、陶土、海泡石粉和沸石粉的至少一种。(The invention relates to granules, a preparation method and application thereof. The granules are spheres with the particle size of 0.2 to 2mm and comprise an active component, a first component, a second component, a third component and a fourth component; the active component is bacillus thuringiensis; the first component is at least one of polyvinyl alcohol, sodium carboxymethyl cellulose, methyl cellulose, xanthan gum, sodium polyacrylate, soluble starch and gelatin; the second component is at least one of ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, pregelatinized starch, cross-linked polyvinylpyrrolidone and cross-linked sodium polyacrylate; the third component is at least one of sodium dodecyl benzene sulfonate, alkyl naphthalene sulfonate formaldehyde condensate, sodium lignin sulfonate, sodium lauryl polyoxyethylene ether sulfate and the like; the fourth component is at least one of bentonite, kaolin, attapulgite, diatomite, pottery clay, sepiolite powder and zeolite powder.)

1. A granule which is a sphere having a particle size of 0.2 to 2mm, comprising an active ingredient, a first component, a second component, a third component and a fourth component; wherein the content of the first and second substances,

the active component is one of bacillus thuringiensis raw powder, bacillus thuringiensis crystal protein and bacillus thuringiensis cell crystal mixture;

the first component is at least one of polyvinyl alcohol, sodium carboxymethylcellulose, methylcellulose, xanthan gum, sodium polyacrylate, soluble starch and gelatin;

the second component is at least one of ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, pregelatinized starch, cross-linked polyvinylpyrrolidone and cross-linked sodium polyacrylate;

the third component is at least one of sodium dodecyl benzene sulfonate, alkyl naphthalene sulfonate formaldehyde condensate, sodium lignosulfonate, calcium lignosulfonate, sodium laureth sulfate, alkyl naphthalene formaldehyde condensate sulfonate, sodium dodecyl sulfate, potassium oleate, sodium oleate, alkyl polyoxyethylene ether sulfonate, alkylphenol polyoxyethylene ether formaldehyde condensate, long-chain polycarboxylate and acrylic acid-sulfonate copolymer;

the fourth component is at least one of bentonite, kaolin, attapulgite, diatomite, pottery clay, sepiolite powder and zeolite powder;

preferably, the granules are spheres having a particle size of 0.27 to 0.55 mm.

2. The granule according to claim 1, wherein the active ingredient is present in an amount of 5% or more, the first component is present in an amount of 0.3% to 1%, the second component is present in an amount of 0.1% to 10%, the third component is present in an amount of 0.1% to 10%, and the balance is the fourth component, based on 100% of the total mass of the granule;

preferably, the content of the active component is 10% to 20%;

preferably, the content of the first component is 0.5% to 0.8%.

3. A granule according to claim 1 or 2, further comprising a warning colour.

4. A granule according to claim 3, wherein the warning colour is at least one of methyl violet, bromophenol blue, carmine, congo red and rose bengal.

5. A granule according to claim 3, wherein the warning color is present in an amount of 0.01 to 3% based on 100% by mass of the total mass of the granule.

6. A process for preparing a granulate according to any one of claims 1 to 5, comprising the steps of:

1) uniformly mixing the active component, the second component, the third component and the fourth component, and crushing to obtain a first mixture;

2) mixing the first component with water to obtain a second mixture;

3) and putting the first mixture into a granulator, uniformly spraying the second mixture onto the first mixture, granulating, and drying to obtain the granules.

7. A method as claimed in claim 6, wherein the granulator is a disk granulator and the second mixture is sprayed onto the first mixture uniformly while the disk is rotating.

8. The method of claim 6, wherein the particle size of the first mixture is 250 to 600 mesh; the content of the first component in the second mixture is 0.3% to 5%; preferably, the content of the first component in the second mixture is 1% to 2%.

9. Use of a granulate according to any of claims 1 to 5 or prepared by a process according to any of claims 6 to 8 for protection against uv light.

10. Use of a granulate according to any one of claims 1 to 5 or prepared by a process according to any one of claims 6 to 8 for combating corn pests, in particular spodoptera frugiperda.

Technical Field

The invention relates to a granule, in particular to an agricultural anti-ultraviolet granule applied to corn.

Background

Agricultural pests are important factors for limiting high and stable yield of crops. The food ingested by the pest accounts for about 1/4 of the total yield each year. Chemical pesticides have been expected for a long time, but problems of environmental pollution, increased residual toxicity, poisoning of people and livestock and the like are faced when the chemical pesticides are singly used, the drug resistance of pests is also increased linearly, the drug concentration is increased continuously, and the prevention and treatment cost is increased continuously. While preventing and controlling pests, the pesticide also kills natural enemies and destroys ecological balance.

Bacillus thuringiensis (Bt for short) is a gram-positive bacterium widely distributed in nature, and is characterized in that parasporal crystallin can be produced in the process of spore formation. Parasporal crystallins are composed of one or more proteins with highly specific insecticidal activity, such proteins are commonly referred to as insecticidal crystallins. The parasympathetic crystallin exists in a protoxin state, and after it enters the digestive tract of a sensitive insect, it is dissolved in an alkaline environment and activated by protease into a toxic polypeptide, which binds to a receptor on the striatal membrane of the intestinal tract in the insect and forms a pore on the cell membrane, disrupting the osmotic balance of the cell, causing cell lysis, and finally leading to death of the larva. Therefore, as a new technical means for preventing and controlling agricultural and forestry pests, a series of microbial pesticide preparations represented by bacillus thuringiensis are widely regarded in the world.

At present, Bt pesticides sold in the market are mainly prepared by enriching fermentation products in Bt fermentation liquor and then preparing preparations, various components such as Bt spores, parasporal crystals, nutrients and the like are mixed, wherein the parasporal crystals have low content and poor stability, and when the Bt pesticides are used in fields, the Bt preparations are easy to cause unstable pesticide effect and short duration due to the problems of ultraviolet radiation, rain wash and the like, so that the popularization and the use of the biological pesticides are influenced.

Disclosure of Invention

One of the present invention provides a granule which is a sphere having a particle size of 0.2 to 2mm, comprising an active component, a first component, a second component, a third component and a fourth component; wherein the content of the first and second substances,

the active component is one of bacillus thuringiensis raw powder, bacillus thuringiensis crystal protein and bacillus thuringiensis cell crystal mixture;

the first component is at least one of polyvinyl alcohol, sodium carboxymethylcellulose, methylcellulose, xanthan gum, sodium polyacrylate, soluble starch and gelatin;

the second component is at least one of ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, cellulose acetate, pregelatinized starch, cross-linked polyvinylpyrrolidone and cross-linked sodium polyacrylate;

the third component is at least one of sodium dodecyl benzene sulfonate, alkyl naphthalene sulfonate formaldehyde condensate, sodium lignosulfonate, calcium lignosulfonate, sodium laureth sulfate, alkyl naphthalene formaldehyde condensate sulfonate, sodium dodecyl sulfate, potassium oleate, sodium oleate, alkyl polyoxyethylene ether sulfonate, alkylphenol polyoxyethylene ether formaldehyde condensate, long-chain polycarboxylate and acrylic acid-sulfonate copolymer;

the fourth component is at least one of bentonite, kaolin, attapulgite, diatomite, pottery clay, sepiolite powder and zeolite powder.

In one embodiment, the granules are spheres having a particle size of 0.27 to 0.55 mm.

In one embodiment, the active ingredient is present in an amount of 5% or more, the first component is present in an amount of 0.3% to 1%, the second component is present in an amount of 0.1% to 10%, the third component is present in an amount of 0.1% to 10%, and the balance is the fourth component, based on 100% of the total mass of the granule.

In one embodiment, the active ingredient is present in an amount of 10% to 20%.

In one embodiment, the first component is present in an amount of 0.5% to 0.8%.

In a specific embodiment, the granules further comprise a warning color.

In a specific embodiment, the warning color is at least one of methyl violet, bromophenol blue, carmine, congo red, and rose bengal.

In a specific embodiment, the warning color is contained in an amount of 0.01 to 3% by mass based on 100% by mass of the total mass of the granule.

The second aspect of the invention provides a process for the preparation of a granulate according to any one of the first to the second aspects of the invention, comprising the steps of:

1) uniformly mixing the active component, the second component, the third component and the fourth component, and crushing to obtain a first mixture;

2) mixing the first component with water to obtain a second mixture;

3) and putting the first mixture into a granulator, uniformly spraying the second mixture onto the first mixture, granulating, and drying to obtain the granules.

In a particular embodiment, the first component is present in the second mixture in an amount of 1% to 2%.

In one embodiment, the pelletizer is a disk pelletizer, and the second mixture is uniformly sprayed onto the first mixture while the disk is rotating.

In one embodiment, the particle size of the first mixture is 250 to 600 mesh; the first component is present in the second mixture in an amount of 0.3% to 5%.

The third invention provides the use of the granules according to any one of the first invention or the granules prepared by the method of any one of the second invention in ultraviolet resistance.

The fourth invention provides the application of the granules in one aspect of the invention or the granules prepared by the method in the second aspect of the invention in preventing and controlling corn pests, in particular spodoptera frugiperda.

The invention has the beneficial effects that:

the invention firstly discovers that the granules prepared from the components of the invention can effectively protect the active components from being rapidly degraded due to the irradiation of ultraviolet rays.

It is worth mentioning that when the granules provided by the invention are used for controlling crop pests, the technical effect of controlling the pests is not completely consistent with the anti-ultraviolet technical effect of the granules provided by the invention, but the particle size of the granules can produce unexpected technical effect on controlling the pests. Specifically, although the invention does not show the experimental results of the granules with the particle sizes of more than 2mm and less than 0.2mm, the experimental results prove that the data results of the granules with the particle sizes of more than 2mm and less than 0.2mm are significantly worse than the data results of the embodiments of the invention, especially the results are worse when the particle sizes are less than 0.2mm, and the technical effect of controlling pests brought by the particle sizes of the embodiments of the invention is combined, so that the selection of the particle sizes of the granules plays an important role in playing the activity of microbial pesticides, especially bacillus thuringiensis, the control of the pests is not facilitated when the particle sizes are too large or too small, and the control of the pests is greatly facilitated when the granules with the proper particle sizes are selected.

Drawings

FIG. 1 shows the amount of Cry1Aa before and after UV treatment.

Detailed Description

The above-described aspects of the invention are explained in more detail below by means of preferred embodiments, but they are not intended to limit the invention.

The reagents in the examples of the present invention were all commercially available unless otherwise specified.

The raw powder of G033A was produced by Wuhan Kenuo Biotech GmbH.

Dissolving a small amount of G033A raw powder in 20mM Na₂CO₃-NaHCO₃SDS-PAGE was performed in aqueous solution (pH 9.8) and quantitated with BSA. The results showed that the mass content of crystallin in the raw powder of G033A was 10%.

Example 1

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G ethyl cellulose, 40G pregelatinized starch, 10G sodium lauryl sulfate, 5G rose bengal and 815G sepiolite powder were mixed, pulverized to 325 mesh, and put into a disk pelletizer.

2) 10g of polyvinyl alcohol 17-88 was dissolved in 190mL of water to prepare a 5% aqueous solution.

3) Gradually spraying 5% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 10-mesh sieve, sieving the granules below 10 meshes with a 20-mesh sieve to obtain granules below 10 meshes and above 20 meshes, and drying to obtain granules with the particle size of 0.8-1.7 mm.

Example 2

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G ethyl cellulose, 40G pregelatinized starch, 10G sodium lauryl sulfate, 5G rose bengal and 817G sepiolite powder were mixed, pulverized to 325 mesh, and put into a disk pelletizer.

2) 8g of polyvinyl alcohol 17-88 was dissolved in 392mL of water to prepare a 2% aqueous solution.

3) Gradually spraying 2% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 40-mesh sieve to obtain granules below 30 meshes and above 40 meshes, and drying to obtain granules with the particle size of 0.38-0.55 mm.

Example 3

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G ethyl cellulose, 40G pregelatinized starch, 10G sodium lauryl sulfate, 5G rose bengal and 820G sepiolite powder were mixed, pulverized to 325 mesh, and put into a disk pelletizer.

2) 5g of polyvinyl alcohol 17-88 was dissolved in 495mL of water to prepare a 1% aqueous solution.

3) Gradually spraying 1% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 50-mesh sieve to obtain granules below 30 meshes and above 50 meshes, and drying to obtain granules with the particle size of 0.27-0.55 mm.

Example 4

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G ethyl cellulose, 40G pregelatinized starch, 10G sodium lauryl sulfate, 5G rose bengal and 913.5G sepiolite powder were mixed, pulverized to 325 mesh, and put into a disk pelletizer.

2) 1.5g of polyvinyl alcohol 17-88 was dissolved in 498 water to make a 0.3% aqueous solution.

3) Gradually spraying 0.3% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 60-mesh sieve, sieving the granules below 60 meshes with a 80-mesh sieve to obtain granules below 60 meshes and above 80 meshes, and drying to obtain granules with the particle size of 0.18-0.25 mm.

Example 5

Granule of 10% Bt G033A

1) 100G033A raw powder, 0.5G ethyl cellulose, 0.5G pregelatinized starch, 1G sodium lauryl sulfate, 0.1G rose bengal and 892.9G sepiolite powder were mixed, pulverized to 250 mesh, and put into a disk pelletizer.

2) 5g of polyvinyl alcohol 17-88 was dissolved in 495mL of water to prepare a 1% aqueous solution.

3) Gradually spraying 1% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 50-mesh sieve to obtain granules below 30 meshes and above 50 meshes, and drying to obtain granules with the particle size of 0.27-0.55 mm.

Example 6

Granule of 10% Bt G033A

1) 100G033A raw powder, 60G ethyl cellulose, 40G pregelatinized starch, 100G sodium lauryl sulfate, 30G rose bengal and 665G sepiolite powder were mixed, pulverized to 600 mesh, and put into a disk pelletizer.

2) 5g of polyvinyl alcohol 17-88 was dissolved in 495mL of water to prepare a 1% aqueous solution.

3) Gradually spraying 1% polyvinyl alcohol 17-88 water solution onto the powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 50-mesh sieve to obtain granules below 30 meshes and above 50 meshes, and drying to obtain granules with the particle size of 0.27-0.55 mm.

Example 7

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G of crosslinked polyvinylpyrrolidone, 40G of crosslinked sodium polyacrylate, 5G of sodium dodecyl sulfate, 5G of sodium lignosulfonate, 5G of rose bengal and 820G of attapulgite were mixed, pulverized to 325 meshes, and put into a disc granulator.

2) 5g of xanthan gum was dissolved in 495mL of water to make a 1% aqueous solution.

3) Gradually spraying 1% xanthan gum water solution onto powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 50-mesh sieve to obtain granules below 30 meshes and above 50 meshes, and drying to obtain granules with the particle size of 0.27-0.55 mm.

Example 8

Granule of 10% Bt G033A

1) 100G033A raw powder, 20G of cross-linked polyvinylpyrrolidone, 40G of cross-linked sodium polyacrylate, 10G of lauryl alcohol polyoxyethylene ether sodium sulfate, 5G of rose bengal and 820G of attapulgite are mixed, crushed to 325 meshes and put into a disc granulator.

2) 5g of soluble starch was dissolved in 495mL of water to make a 1% aqueous solution.

3) Gradually spraying 1% soluble starch water solution onto powder in a disc granulator under the rotation state of a disc, sieving the obtained granules with a 30-mesh sieve, sieving the granules below 30 meshes with a 50-mesh sieve to obtain granules below 30 meshes and above 50 meshes, and drying to obtain granules with the particle size of 0.27-0.55 mm.

Comparative example 1

G033A raw powder.

Comparative example 2

Respectively sieving attapulgite, sodium dodecyl sulfate, sodium lignin sulfonate and xanthan gum with a 400-mesh standard sieve, adding 5g of sodium dodecyl sulfate, 5g of sodium lignin sulfonate and 5g of xanthan gum into 875g of attapulgite, uniformly stirring, then adding 100g of dried bacillus thuringiensis raw drug into the mixture, uniformly mixing and processing into mixed wettable powder.

Ultraviolet irradiation

50mg of the products obtained in examples 1 to 8 and comparative example 2 and 5mg of the raw G033A powder in comparative example 1 were added to 10ml of ddH₂After O, diluted, the amount of the Cry1Aa protein in each product was measured by ELISA method using Cry 1-type antibody, and then the average value of the amount (mg/mL) of the Cry1Aa protein in each product of examples 1 to 8 and comparative examples 1 and 2 in water was calculated and used as a Control (CK).

50mg of the products obtained in examples 1 to 8 and comparative example 2, 5mg of the raw G033A powder of comparative example 1 were placed in a 9cm disposable plastic dish at an ultraviolet intensity of 240. mu.w/cm²The ultraviolet irradiation was performed 1 time per 4 hours with an interval of 0.5 hours, 3 times for a total of 12 hours, respectively, under a 30cm ultraviolet lamp of (1) to obtain ultraviolet-irradiated samples of each example or comparative example. The UV-irradiated samples of each example and comparative example were added with 10ml of ddH₂After O, diluting, and detecting the content (mg/mL) of the Cry1Aa protein in water in each ultraviolet irradiation sample by an ELISA method by using Cry1 antibodies.

The results are shown in FIG. 1.

As can be seen from the results of fig. 1, the uv resistance effect of the active ingredients in the granules prepared in examples 1 to 8 was significantly superior to that of the comparative example; however, in the granules prepared in each example, there was a significant difference in the ultraviolet resistance, except for the granules prepared in examples 5 and 6, that is, the granule prepared in example 1 > the granule prepared in example 2 > the granule prepared in example 3 > the granule prepared in example 5 ≈ the granule prepared in example 6 > the granule prepared in example 4 > the granule prepared in example 7 > the granule prepared in example 8.

Experiment in the field

The test area is 1 mu in total, each treatment is repeated for 3 times, and a plot without drug application is used as a blank control. The control object is corn crop pest Spodoptera frugiperda.

The application period is as follows: in the corn jointing stage, the average height of corn plants is about 40cm, and the corn density is about 4000 plants/mu.

The application method comprises the following steps: examples 1 to 10 and comparative example 2 an agricultural unmanned aerial vehicle anyang quanfeng shin ZP equipped with a spraying system and a particle sowing system was used. The spreading amount of the granules is 1 Kg/mu. The spraying amount of the wettable powder is 1 Kg/mu. Wherein, the flying height of the agricultural unmanned aerial vehicle particles is 3m when the agricultural unmanned aerial vehicle particles are spread, and the flying speed is 3 m/S; the flying height of the agricultural unmanned aerial vehicle is 2m when the agricultural unmanned aerial vehicle sprays, and the flying speed is 3 m/S. The raw powder of comparative example 1 was uniformly sprayed using a hand-held knapsack sprayer (100g G033A raw powder/30L water/acre).

The investigation method comprises the following steps: investigating population cardinality of spodoptera frugiperda larvae before application, investigating the latest 2 leaf damage conditions 7d after application, investigating 50 plants in each cell, scoring (0-9 points) the investigated plant damage according to the Davis damage grading standard of Table 2, and calculating the control effect according to the Davis index. The results are shown in Table 3.

TABLE 2

TABLE 3

Examples	Number of insects on day of drug administration (head/50 plants)	Control effect (%) (7 days after drug administration)
			Example 1	167±23	68.7±2.4c
Example 2	158±18	85.6±4.9a
			Example 3	179±15	87.4±5.9a
Example 4	165±15	76.6±3.5b
			Example 7	172±21	60.7±5.1d
Comparative example 1	180±9	56.4±5.2d
			Comparative example 2	179±14	60.5±2.2d
Blank control	162±11	——e