A composition for preventing and treating anthracnose of pepper and a preparation method thereof

Abstract

This invention provides a composition for controlling anthracnose in peppers and its preparation method, belonging to the field of plant disease control technology. The composition of this invention, by weight, comprises the following components: 30-55 parts of Bacillus fermentation culture broth, 5-17 parts of loquat leaf extract, 3-8 parts of hawthorn leaf extract, 2-6 parts of milk thistle extract, 1-5 parts of sodium aluminosilicate, 4-7 parts of polyethylene glycol, and 5-8 parts of ε-polylysine. This invention, by mixing Bacillus fermentation culture broth, compound plant extracts, sodium aluminosilicate, polyethylene glycol, and ε-polylysine, can effectively control anthracnose in peppers. The addition of sodium aluminosilicate, polyethylene glycol, and ε-polylysine to the formula prevents flocculation and precipitation, and improves the storage stability of the composition.

Description

Technical Field

[0001] This invention relates to the field of plant disease control technology, specifically to a composition for controlling anthracnose in peppers and its preparation method. Background Technology

[0002] Anthracnose, commonly known as ring spot or ring rot, is a major disease affecting the growth of chili peppers and sweet peppers, caused by fungi of the genera *Colletotrichum* and *Discocephalum*. It primarily damages the fruit and leaves of nearly mature peppers, leading to fruit rot, leaf drop, and reduced yield, with mature fruits being most severely affected. Anthracnose is prone to outbreaks in hot and humid environments and is a significant limiting disease in chili-producing areas of my country. Based on the color of the lesions on the affected fruit, anthracnose is generally classified into black spot anthracnose, black anthracnose, and red anthracnose. Black spot anthracnose mainly affects mature fruits, with larger, darker black spots that ooze a sticky substance under humid conditions. Black anthracnose can affect both leaves and fruits, with irregular, sunken lesions featuring slightly raised concentric rings, numerous small black dots, and a moist, discolored ring around the perimeter. Red anthracnose can affect both mature and young fruits. The lesions appear as water-soaked, yellowish-brown circular spots that are sunken. Small orange-red dots grow on the lesions in concentric rings. Under humid conditions, a light red sticky substance will ooze from the surface of the entire lesion.

[0003] Currently, the main method for controlling anthracnose in peppers is chemical control. To reduce the occurrence of anthracnose on the fruit, pepper growers usually apply chemical agents 1-2 days before harvest. This leaves a large amount of fungicide residue on the surface of the harvested peppers, thus preventing damage from the anthracnose pathogen. However, while this method achieves the effect of controlling anthracnose, the residual chemical fungicides on the pepper surface pose risks such as carcinogenicity and teratogenicity, seriously affecting human health. Therefore, providing a safe and effective composition for controlling anthracnose in peppers is of great significance. Summary of the Invention

[0004] In view of this, the present invention provides a composition for preventing and controlling anthracnose in peppers, comprising, by weight, the following components:

[0005] 30-55 parts of Bacillus fermentation broth, 5-17 parts of loquat leaf extract, 3-8 parts of hawthorn leaf extract, 2-6 parts of milk thistle extract, 1-5 parts of sodium aluminosilicate, 4-7 parts of polyethylene glycol, and 5-8 parts of ε-polylysine.

[0006] Furthermore, the method for preparing the Bacillus fermentation culture broth includes the following steps:

[0007] A culture medium was prepared using maltose, ammonium sulfate, soybean meal powder, magnesium sulfate, and dipotassium hydrogen phosphate as raw materials. Bacillus licheniformis was inoculated into the culture medium and fermented to obtain a Bacillus fermentation culture broth.

[0008] Furthermore, the fermentation conditions include: fermentation temperature 30-37℃, pH 6.5-7.5, rotation speed 200-300 r / min, and fermentation time 36-48 h.

[0009] Furthermore, the maltose has a mass concentration of 1-3 g / L, the ammonium sulfate has a mass concentration of 3-8 g / L, the soybean meal has a mass concentration of 5-12 g / L, the magnesium sulfate has a mass concentration of 7-10 g / L, the dipotassium hydrogen phosphate has a mass concentration of 1-2 g / L, and the Bacillus licheniformis inoculation amount is 1-2%.

[0010] Furthermore, the viable count of Bacillus licheniformis in the Bacillus fermentation culture broth is 1.5 × 10⁻⁶. 9 cfu / g.

[0011] Furthermore, the method for preparing the loquat leaf extract includes the following steps: pulverizing the collected dried loquat leaves, sieving them to obtain loquat leaf powder; firstly, extracting the loquat leaf powder by reflux with 75% ethanol, filtering, collecting the residue and filtrate, extracting the residue by reflux with n-hexane, combining the two filtrates, concentrating under reduced pressure to recover the solvent, and drying to obtain the loquat leaf extract.

[0012] Furthermore, the ethanol reflux extraction is performed for 2-3 hours, the n-hexane reflux extraction is performed for 1-1.5 hours, the ratio of loquat leaf powder to 75% ethanol is 1:8-10, the ratio of filter residue to n-hexane is 1:5-7, and the drying temperature is 60-75℃.

[0013] Furthermore, the preparation method of the hawthorn leaf extract includes the following steps:

[0014] The dried hawthorn leaves were pulverized and sieved to obtain hawthorn leaf powder. Water was added to the hawthorn leaf powder and decocted twice. The filtrates were combined, concentrated under reduced pressure, and dried to obtain hawthorn leaf extract.

[0015] Furthermore, the first decoction time is 2-3 hours with a material-to-liquid ratio of 1:6-8, the second decoction time is 1-2 hours with a material-to-liquid ratio of 1:3-5, the decoction temperature is 100℃, and the drying temperature is 60℃.

[0016] Furthermore, the preparation method of the milk thistle extract includes the following steps:

[0017] Milk thistle is dried, pulverized, and sieved to obtain milk thistle powder. Water is added to the milk thistle powder and soaked for a certain period of time. After soaking, microwave extraction is performed, followed by filtration. The residue and filtrate are collected. Water is added to the residue, and the soaking and microwave extraction processes are repeated. The filtrate is collected twice, concentrated, and dried to obtain milk thistle extract.

[0018] Furthermore, the drying temperature is 50-65℃, the material-to-liquid ratio is 1:6-10, the soaking time is 20-40 minutes, the microwave power is 400-500W, the microwave extraction time is 5-10 minutes, and the drying temperature is 60℃.

[0019] The present invention also provides a method for preparing the composition for preventing and controlling anthracnose in peppers, the method comprising: mixing Bacillus fermentation culture broth, loquat leaf extract, hawthorn leaf extract, milk thistle extract, sodium aluminosilicate, polyethylene glycol, and ε-polylysine in a certain proportion to obtain the composition for preventing and controlling anthracnose in peppers.

[0020] Furthermore, the method of using the composition for controlling anthracnose in peppers includes preparing a mixture with water to a concentration of 2.0-3.8 wt%, applying it to the roots once a week after transplanting the peppers and when no symptoms appear on the leaves, followed by applying it to the roots once every 10 days, for a total of three applications, with an application rate of 30 mL per plant.

[0021] Compared with the prior art, the present invention has the following beneficial effects: The present invention mixes Bacillus fermentation culture medium, compound plant extracts, sodium aluminosilicate, polyethylene glycol and ε-polylysine, and achieves the effect of effectively preventing and controlling anthracnose in peppers through the synergistic effect between the raw materials.

[0022] Furthermore, the inventors discovered during their research that the Bacillus fermentation broth contains a large amount of macromolecules such as proteins and polysaccharides, which are prone to flocculation or precipitation during long-term storage. To address this issue, this invention introduces sodium aluminosilicate, polyethylene glycol, ε-polylysine, and plant extracts. Through the synergistic effect of these raw materials, the dispersibility of the system is improved, particle aggregation is prevented, and the storage stability of the composition is enhanced. Detailed Implementation

[0023] This invention provides a composition for preventing and controlling anthracnose in peppers, comprising the following components by weight: 30-55 parts of Bacillus fermentation culture broth, 5-17 parts of loquat leaf extract, 3-8 parts of hawthorn leaf extract, 2-6 parts of milk thistle extract, 1-5 parts of sodium aluminosilicate, 4-7 parts of polyethylene glycol, and 5-8 parts of ε-polylysine.

[0024] Preferably, the composition for preventing and controlling anthracnose in peppers comprises, by weight, the following components: 50 parts of Bacillus fermentation culture broth, 12 parts of loquat leaf extract, 5 parts of hawthorn leaf extract, 6 parts of milk thistle extract, 3 parts of sodium aluminosilicate, 5 parts of polyethylene glycol, and 7 parts of ε-polylysine.

[0025] Furthermore, the inventors discovered during their research that the Bacillus fermentation broth contains a large amount of macromolecules such as proteins and polysaccharides, which are prone to flocculation or precipitation during long-term storage. To address this issue, this invention introduces sodium aluminosilicate, polyethylene glycol, ε-polylysine, and plant extracts. Sodium aluminosilicate adsorbs macromolecules such as proteins and polysaccharides in the fermentation broth, reducing the probability of their binding and mitigating charge-induced flocculation by regulating the ionic strength in the system. Polyethylene glycol molecules can adsorb onto the surface of macromolecules, preventing interparticle aggregation through steric hindrance. ε-polylysine, through charge regulation, binds to negatively charged proteins and other substances to form stable complexes, reducing aggregation and improving the homogeneity of the composition by lowering interfacial tension. ε-polylysine also possesses antibacterial properties, which can enhance the composition's effectiveness against anthracnose in peppers to some extent. Under low-temperature conditions, the plant extracts synergistically interact with sodium aluminosilicate, polyethylene glycol, and ε-polylysine, improving the stability of the molecular structure and enhancing the antibacterial activity of ε-polylysine.

[0026] In some embodiments of the present invention, the method for preparing the Bacillus fermentation culture broth includes the following steps: preparing a culture medium using maltose, ammonium sulfate, soybean meal powder, magnesium sulfate, and dipotassium hydrogen phosphate as raw materials; inoculating Bacillus licheniformis into the culture medium; and carrying out fermentation culture to obtain the Bacillus fermentation culture broth.

[0027] In some embodiments of the present invention, the fermentation conditions include: fermentation temperature 30-37℃, pH 6.5-7.5, rotation speed 200-300 r / min, and fermentation time 36-48 h. Preferably, the fermentation conditions include: fermentation temperature 35℃, pH 7.5, rotation speed 230 r / min, and fermentation time 36 h. The present invention controls the fermentation conditions within the above ranges to ensure fermentation effectiveness and simultaneously enhance the composition's control effect against anthracnose in peppers.

[0028] In some embodiments of the present invention, the maltose has a mass concentration of 1-3 g / L, the ammonium sulfate has a mass concentration of 3-8 g / L, the soybean meal powder has a mass concentration of 5-12 g / L, the magnesium sulfate has a mass concentration of 7-10 g / L, and the dipotassium hydrogen phosphate has a mass concentration of 1-2 g / L; the inoculum size of Bacillus licheniformis is 1-2%. Preferably, the maltose has a mass concentration of 2 g / L, the ammonium sulfate has a mass concentration of 5 g / L, the soybean meal powder has a mass concentration of 7 g / L, the magnesium sulfate has a mass concentration of 8 g / L, the dipotassium hydrogen phosphate has a mass concentration of 1 g / L, and the inoculum size of Bacillus licheniformis is 1%.

[0029] In some embodiments of the present invention, the viable count of Bacillus licheniformis in the Bacillus fermentation culture broth is 1.5 × 10⁻⁶. 9 cfu / g.

[0030] In some embodiments of the present invention, the method for preparing the loquat leaf extract includes the following steps: pulverizing and sieving the collected dried loquat leaves to obtain loquat leaf powder; firstly, extracting the loquat leaf powder by reflux with 75% ethanol, filtering, collecting the filter residue and filtrate, extracting the filter residue by reflux with n-hexane, combining the two filtrates, concentrating under reduced pressure to recover the solvent, and drying to obtain the loquat leaf extract.

[0031] In some embodiments of the present invention, the ethanol reflux extraction is performed for 2-3 hours, the n-hexane reflux extraction for 1-1.5 hours, the ratio of loquat leaf powder to 75% ethanol is 1:8-10, the ratio of filter residue to n-hexane is 1:5-7, and the drying temperature is 60-75°C. Preferably, the ethanol reflux extraction is performed for 2 hours, the n-hexane reflux extraction for 1 hour, the ratio of loquat leaf powder to 75% ethanol is 1:8, the ratio of filter residue to n-hexane is 1:5, and the drying temperature is 60°C. The present invention limits the extraction process parameters within the above ranges, which helps to improve extraction efficiency.

[0032] In some embodiments of the present invention, the preparation method of the hawthorn leaf extract includes the following steps:

[0033] The dried hawthorn leaves were pulverized and sieved to obtain hawthorn leaf powder. Water was added to the hawthorn leaf powder and decocted twice. The filtrates were combined, concentrated under reduced pressure, and dried to obtain hawthorn leaf extract.

[0034] In some embodiments of the present invention, the first decoction time is 2-3 hours with a material-to-liquid ratio of 1:6-8, the second decoction time is 1-2 hours with a material-to-liquid ratio of 1:3-5, the decoction temperature is 100°C, and the drying temperature is 60°C. Preferably, the first decoction time is 3 hours with a material-to-liquid ratio of 1:8, the second decoction time is 2 hours with a material-to-liquid ratio of 1:5, the decoction temperature is 100°C, and the drying temperature is 60°C. The present invention limits the extraction process parameters within the above ranges, which helps to improve extraction efficiency.

[0035] In some embodiments of the present invention, the method for preparing the milk thistle extract includes the following steps:

[0036] Milk thistle is dried, pulverized, and sieved to obtain milk thistle powder. Water is added to the milk thistle powder and soaked for a certain period of time. After soaking, microwave extraction is performed, followed by filtration. The residue and filtrate are collected. Water is added to the residue, and the soaking and microwave extraction processes are repeated. The filtrate is collected twice, concentrated, and dried to obtain milk thistle extract.

[0037] In some embodiments of the present invention, the drying temperature is 50-65°C, the material-to-liquid ratio is 1:6-10, the soaking time is 20-40 min, the microwave power is 400-500 W, the microwave extraction time is 5-10 min, and the drying temperature is 60°C. Preferably, the drying temperature is 65°C, the material-to-liquid ratio is 1:7, the soaking time is 20 min, the microwave power is 400 W, the microwave extraction time is 6 min, and the drying temperature is 60°C. The present invention limits the extraction process parameters within the above ranges, which helps to improve extraction efficiency.

[0038] This invention also provides a method for preparing the composition for controlling anthracnose in peppers, the method comprising: mixing Bacillus fermentation culture broth, loquat leaf extract, hawthorn leaf extract, milk thistle extract, sodium aluminosilicate, polyethylene glycol, and ε-polylysine in a specified ratio until homogeneous, thereby obtaining the composition for controlling anthracnose in peppers. This invention does not impose any particular limitation on the mixing method; homogeneous mixing of the components can be achieved using techniques well-known in the art.

[0039] In some embodiments of the present invention, the method of using the composition for controlling anthracnose in peppers involves preparing a mixture with water to a concentration of 2.0-3.8%, and then drenching the roots once a week after transplanting the peppers, when no disease symptoms appear on the leaves. This is repeated three times, with a dosage of 30 mL per plant. The present invention does not impose any special restrictions on the timing of root drenching. However, to improve the control effect, it is recommended to perform the drenching on sunny days between 9-11 am or 3-5 pm, avoiding rainy days, periods of high or low temperature.

[0040] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0041] Unless otherwise specified, the test methods or experimental methods described in the following examples are all conventional methods; unless otherwise specified, the raw materials and additives are obtained from conventional commercial sources or prepared by conventional methods.

[0042] The sodium aluminosilicate, CAS No.: 1344-00-9, was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0043] The polyethylene glycol is polyethylene glycol 400, CAS number: 25322-68-3, purchased from Jinan Xinke Chemical Co., Ltd.

[0044] The ε-polylysine, CAS No.: 28211-04-3, was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0045] Imazalil was purchased from Dongguan Ruidefeng Biotechnology Co., Ltd.

[0046] Bacillus licheniformis XGY13-2 is deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC NO.8974.

[0047] The method for preparing the Bacillus fermentation culture medium is as follows:

[0048] A culture medium was prepared using 2 g / L maltose, 5 g / L ammonium sulfate, 7 g / L soybean meal, 8 g / L magnesium sulfate, and 1 g / L dipotassium hydrogen phosphate as raw materials. *Bacillus licheniformis* was inoculated into the culture medium at an inoculum size of 1%. Fermentation was carried out at 35°C, pH 7.5, and a rotation speed of 230 r / min for 36 h to obtain a *Bacillus licheniformis* fermentation broth. The viable count of *Bacillus licheniformis* in the fermentation broth was 1.5 × 10⁻⁶. 9 cfu / g.

[0049] The preparation method of the loquat leaf extract is as follows: Collected dried loquat leaves are pulverized and passed through a 20-mesh sieve to obtain loquat leaf powder; the loquat leaf powder is first extracted with 75% ethanol under reflux for 2 hours at a material-to-liquid ratio of 1:8, filtered, and the residue and filtrate are collected; the residue is then extracted with n-hexane under reflux for 1 hour at a material-to-liquid ratio of 1:5; the two filtrates are combined, concentrated under reduced pressure to recover the solvent, and dried at 80℃ to obtain the loquat leaf extract.

[0050] The preparation method of the hawthorn leaf extract is as follows:

[0051] The dried hawthorn leaves were pulverized and passed through a 20-mesh sieve to obtain hawthorn leaf powder. Water was added to the hawthorn leaf powder and decocted twice. The first decoction lasted for 3 hours with a material-to-liquid ratio of 1:8, and the second decoction lasted for 2 hours with a material-to-liquid ratio of 1:5. The decoction temperature was 100℃. The filtrates were combined, concentrated under reduced pressure, and dried at 80℃ to obtain hawthorn leaf extract.

[0052] The method for preparing the milk thistle extract is as follows:

[0053] Milk thistle was dried in an oven at 65°C, then pulverized and passed through a 20-mesh sieve to obtain milk thistle powder. Water was added to the milk thistle powder at a material-to-liquid ratio of 1:7, and the mixture was soaked for 20 minutes. After soaking, it was extracted by microwave at 400W for 6 minutes, filtered, and the residue and filtrate were collected. Water was added to the residue again, and the soaking and microwave extraction processes were repeated. The filtrates were collected twice, concentrated, and dried at 80°C to obtain milk thistle extract.

[0054] Example 1

[0055] A method for preparing a composition for preventing and controlling anthracnose in peppers, comprising the following specific steps:

[0056] A composition for preventing and controlling anthracnose in peppers is obtained by mixing 50 parts of Bacillus fermentation culture broth, 12 parts of loquat leaf extract, 5 parts of hawthorn leaf extract, 6 parts of milk thistle extract, 3 parts of sodium aluminosilicate, 5 parts of polyethylene glycol, and 7 parts of ε-polylysine in a certain proportion.

[0057] Example 2

[0058] A method for preparing a composition for preventing and controlling anthracnose in peppers, comprising the following specific steps:

[0059] A composition for preventing and controlling anthracnose in peppers is obtained by mixing 30 parts of Bacillus fermentation culture broth, 5 parts of loquat leaf extract, 3 parts of hawthorn leaf extract, 2 parts of milk thistle extract, 1 part of sodium aluminosilicate, 4 parts of polyethylene glycol, and 5 parts of ε-polylysine in a certain proportion.

[0060] Example 3

[0061] A method for preparing a composition for preventing and controlling anthracnose in peppers, comprising the following specific steps:

[0062] A composition for preventing and controlling anthracnose in peppers is obtained by mixing 55 parts of Bacillus fermentation culture broth, 17 parts of loquat leaf extract, 8 parts of hawthorn leaf extract, 6 parts of milk thistle extract, 5 parts of sodium aluminosilicate, 7 parts of polyethylene glycol, and 8 parts of ε-polylysine in a certain proportion.

[0063] Comparative Example 1

[0064] The difference from Example 1 is that no Bacillus fermentation broth was added, while the rest of the steps are the same as in Example 1.

[0065] Comparative Example 2

[0066] The difference from Example 1 is that loquat leaf extract, hawthorn leaf extract, and milk thistle extract were not added; the remaining steps are the same as in Example 1.

[0067] Comparative Example 3

[0068] The difference from Example 1 is that hawthorn leaf extract was not added, but the rest of the steps are the same as in Example 1.

[0069] Comparative Example 4

[0070] The difference from Example 1 is that milk thistle extract was not added, but the rest of the steps are the same as in Example 1.

[0071] Comparative Example 5

[0072] The difference from Example 1 is that loquat leaf extract was not added, but the rest of the steps are the same as in Example 1.

[0073] Performance testing

[0074] (a) The compositions provided in Examples 1-3 and Comparative Examples 1-5 were used to control anthracnose in peppers. The test methods are as follows:

[0075] Experimental area: Chili pepper growing area in Ninghe District, Tianjin. It has a warm temperate monsoon continental climate with four distinct seasons, abundant sunshine, an average annual temperature of 11.1℃, an average annual ground temperature of 2.7℃~4.9℃, an average annual sunshine duration of 2801.7 hours, an annual sunshine percentage of 63%, an average annual precipitation of 637 mm, a frost-free period of 240 days, and a flat terrain with an elevation of about 1.65-2.0 meters.

[0076] Experimental method: The selected chili pepper planting areas were randomly divided into 8 experimental areas, 1 blank control area, and 1 positive control area.

[0077] The compositions provided in Examples 1-3 and Comparative Examples 1-5 were diluted with water to prepare a 3.0% mixture. One week after transplanting, when no disease symptoms appeared on the leaves, the mixture was applied as a root drench. This was repeated three times, with a dosage of 30 mL per plant. The compositions provided in Examples 1-3 and Comparative Examples 1-5 were applied to eight experimental areas, respectively. An equal volume of water was applied to the blank control area, and an equal volume of 0.2 g / L prochloraz was applied to the positive control area. Field management was carried out according to conventional chili pepper cultivation methods. Each treatment was replicated five times, and the average value was taken. Thirty days after the application of the pesticides, the incidence of anthracnose in each area was counted, and the results are shown in Table 1.

[0078] Group Anthrax incidence Blank control group 62.05% Example 1 0.15% Example 2 0.38% Example 3 0.21% Comparative Example 1 31.60% Comparative Example 2 49.25% Comparative Example 3 16.41% Comparative Example 4 10.85% Comparative Example 5 12.32% Positive control group 0.81%

[0079] As shown in Table 1, compared with Comparative Examples 1-5, the compositions prepared in Examples 1-3 are more effective in controlling anthracnose in peppers. Furthermore, the control effect of the compositions in Examples 1-3 is superior to commonly used chemical agents. This indicates that the compositions provided by the present invention can effectively control anthracnose in peppers and have the potential to replace chemical agents. This provides support for the research and development of safe, environmentally friendly, and highly efficient biological pesticides for controlling anthracnose in peppers, and is of great significance to the sustainable development of the pepper industry. In addition, the dosage of the anthracnose control compositions provided by the present invention is relatively small, which helps to reduce the cost of controlling anthracnose in peppers.

[0080] Comparative Example 6

[0081] The difference from Example 1 is that sodium aluminosilicate was not added, but the rest of the steps are the same as in Example 1.

[0082] Comparative Example 7

[0083] The difference from Example 1 is that polyethylene glycol was not added, but the rest of the steps are the same as in Example 1.

[0084] Comparative Example 8

[0085] The difference from Example 1 is that ε-polylysine was not added, while the rest of the steps are the same as in Example 1.

[0086] (ii) The compositions provided in Examples 1-3 and Comparative Examples 6-8 were mixed with water to prepare a 200 mL mixture with a concentration of 3%, which was then sealed and stored in the dark at an ambient temperature of 25±2℃. The state of the mixture was observed at the 1st month, 3rd month, 6th month, 9th month and 12th month, and the results are shown in Table 2.

[0087] Table 2

[0088] Group Month 1 3rd month 6th month 9th month 12th month Example 1 Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Example 2 Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Example 3 Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Comparative Example 6 Clear, without flocculation or sedimentation Small amount of flocculation, small amount of sedimentation Small amount of flocculation, small amount of sedimentation More flocculation, less sedimentation More flocculation, more sedimentation Comparative Example 7 Clear, without flocculation or sedimentation Clear, without flocculation or sedimentation Small amount of flocculation, no sedimentation Small amount of flocculation, small amount of sedimentation More flocculation, less sedimentation Comparative Example 8 Clear, without flocculation or sedimentation A small amount of flocculation, no amount of sedimentation Small amount of flocculation, small amount of sedimentation Small amount of flocculation, small amount of sedimentation More flocculation, less sedimentation

[0089] As shown in Table 2, the use of sodium aluminosilicate, polyethylene glycol, and ε-polylysine plays an important role in improving the stability of the composition in this invention. Furthermore, the storage stability of the compositions provided in Examples 1-3 and Comparative Examples 2 and 6-8 was determined at -10°C. The results showed that the compositions of Examples 1-3 did not exhibit flocculation or precipitation after 12 months of storage. However, the compositions prepared in Comparative Examples 2 and 6-8 all showed flocculation and precipitation by the third month of testing, indicating that the use of plant extracts, sodium aluminosilicate, polyethylene glycol, and ε-polylysine plays a crucial role in improving the stability of the composition under low-temperature conditions.

[0090] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition for preventing and controlling anthracnose in peppers, characterized in that, By weight, it includes the following components: 30-55 parts of Bacillus fermentation broth, 5-17 parts of loquat leaf extract, 3-8 parts of hawthorn leaf extract, 2-6 parts of milk thistle extract, 1-5 parts of sodium aluminosilicate, 4-7 parts of polyethylene glycol, and 5-8 parts of ε-polylysine.

2. The composition for controlling anthracnose in peppers according to claim 1, characterized in that, The method for preparing the Bacillus fermentation culture broth includes the following steps: A culture medium was prepared using maltose, ammonium sulfate, soybean meal powder, magnesium sulfate, and dipotassium hydrogen phosphate as raw materials. Bacillus licheniformis was inoculated into the culture medium and fermented to obtain a Bacillus fermentation culture broth.

3. The composition for controlling anthracnose in peppers according to claim 2, characterized in that, The maltose has a mass concentration of 1-3 g / L, the ammonium sulfate has a mass concentration of 3-8 g / L, the soybean meal has a mass concentration of 5-12 g / L, the magnesium sulfate has a mass concentration of 7-10 g / L, the dipotassium hydrogen phosphate has a mass concentration of 1-2 g / L, and the Bacillus licheniformis inoculation amount is 1-2%.

4. The composition for controlling anthracnose in peppers according to claim 1, characterized in that, The method for preparing the loquat leaf extract includes the following steps: pulverizing and sieving the collected dried loquat leaves to obtain loquat leaf powder; firstly, extracting the loquat leaf powder by reflux with 75% ethanol, filtering, collecting the filter residue and filtrate, extracting the filter residue by reflux with n-hexane, combining the two filtrates, concentrating under reduced pressure to recover the solvent, and drying to obtain the loquat leaf extract.

5. The composition for controlling anthracnose in peppers according to claim 4, characterized in that, The ethanol reflux extraction is carried out for 2-3 hours, the n-hexane reflux extraction is carried out for 1-1.5 hours, the ratio of loquat leaf powder to 75% ethanol is 1:8-10, the ratio of filter residue to n-hexane is 1:5-7, and the drying temperature is 60-75℃.

6. The composition for controlling anthracnose in peppers according to claim 1, characterized in that, The preparation method of the hawthorn leaf extract includes the following steps: The dried hawthorn leaves were pulverized and sieved to obtain hawthorn leaf powder. Water was added to the hawthorn leaf powder and decocted twice. The filtrates were combined, concentrated under reduced pressure, and dried to obtain hawthorn leaf extract.

7. The composition for controlling anthracnose in peppers according to claim 1, characterized in that, The method for preparing the milk thistle extract includes the following steps: Milk thistle is dried, pulverized, and sieved to obtain milk thistle powder. Water is added to the milk thistle powder and soaked for a certain period of time. After soaking, microwave extraction is performed, followed by filtration. The residue and filtrate are collected. Water is added to the residue, and the soaking and microwave extraction processes are repeated. The filtrate is collected twice, concentrated, and dried to obtain milk thistle extract.

8. The composition for controlling anthracnose in peppers according to claim 7, characterized in that, The drying temperature is 50-65℃, the material-to-liquid ratio is 1:6-10, the soaking time is 20-40 minutes, the microwave power is 400-500W, the microwave extraction time is 5-10 minutes, and the drying temperature is 60℃.

9. A method for preparing the composition for controlling anthracnose in peppers according to any one of claims 1-8, characterized in that, A composition for preventing and controlling anthracnose in peppers is obtained by mixing Bacillus fermentation culture broth, loquat leaf extract, hawthorn leaf extract, milk thistle extract, sodium aluminosilicate, polyethylene glycol, and ε-polylysine in a certain proportion.

10. The method of using the composition for controlling anthracnose in peppers according to any one of claims 1-8, characterized in that, Prepare a mixture with a concentration of 2.0-3.8% by adding water. Apply the mixture to the roots once a week after transplanting the peppers, when no disease symptoms appear on the leaves. Then apply the mixture to the roots once every 10 days for a total of three applications, with an application rate of 30 mL per plant.