Compound preparation for preventing and treating fungal diseases of tobaccos and use method of compound preparation

Abstract

The invention provides a compound preparation for preventing and treating tobacco fungal diseases and a use method thereof, and belongs to the technical field of tobacco disease prevention and treatment. The composite preparation for preventing and treating the fungal diseases of the tobaccos comprises the following raw materials: quercetin, toosendanin and a composite microbial preparation, and the composite microbial preparation is prepared from 1.5-4.6 billion cfu / g of bacillus subtilis, 0.4-1.5 billion cfu / g of rhizobium and 1.7-3.5 billion cfu / g of bacillus amyloliquefaciens. The compound preparation for preventing and treating the tobacco fungal diseases has the characteristics of prevention and treatment of the tobacco fungal diseases, slow release, long-acting administration, cost reduction, effect improvement, environmental protection and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of tobacco disease control, and particularly relates to a composite preparation for controlling tobacco fungal diseases and a method for using the same. Background Art

[0002] Tobacco (Nicobatiana tabacum) is an annual herbaceous crop, belonging to the class Dicotyledons, the order Tubiflorae, the Solanaceae family, and the genus Nicotiana. It originated in South America and has a history of large-scale cultivation for more than 500 years. Tobacco is an economic crop with a relatively large number of diseases and serious damage. The economic losses caused by tobacco diseases in China are huge every year.

[0003] It is reported that there are about 116 kinds of tobacco diseases in the world, and about 60 kinds of tobacco infectious diseases have been identified in China. Among tobacco diseases, the most numerous and harmful ones are fungal diseases. With the development of the intensive development model of tobacco agricultural production, the cultivation conditions and planting methods of tobacco have been constantly changing. And large-scale planting usually relies on a highly standardized cultivation model. The concentrated planting of single varieties and the extensive use of chemical fertilizers and pesticides have damaged the balance of the soil ecosystem while meeting the high-yield demand. At the same time, the continuous cropping planting method for many years has further exacerbated the deterioration of the soil environment, resulting in a large accumulation of pathogenic bacteria in the soil, and various new and old diseases have occurred one after another, and the tobacco root and stem diseases have become more and more serious. The fungal root and stem diseases of tobacco in China include black shank, root rot, southern blight, gray mold, brown spot, etc.

[0004] The invention patent with the application number CN201710538217.1 discloses a bactericidal composition containing isopyrazam and metalaxyl and its application. The bactericidal composition takes isopyrazam and metalaxyl as the main active ingredients and can be used to control diseases of cereals, fruit trees and vegetables. It has the characteristics of fast bactericidal speed, long-lasting efficacy and low application cost. Although this method can also control tobacco fungal diseases, the disclosed bactericidal composition is mainly composed of chemical agents, and after long-term use, it is prone to disadvantages such as pesticide residues, environmental pollution and enhanced drug resistance. The invention patent with the application number CN201110404047.0 discloses an essential oil agent for controlling tobacco fungal diseases and a method for using the same in the technical field of biological pesticides. The essential oil agent contains: aromatic plant essential oil, emulsifier and dispersant, and its using method is to dilute the essential oil agent with water and then apply it to tobacco. The essential oil agent prepared by this patent has the effect of controlling tobacco fungal diseases. However, the main raw material, plant essential oil, used in it belongs to volatile substances, resulting in poor storage stability of the agent and easy failure.

[0005] Biological control technology has gradually become a hot research direction in the prevention and control of plant diseases due to its advantages such as low cost and environmental friendliness. The principles of biological control of plant diseases include inhibiting harmful bacteria by utilizing the antagonistic relationship formed during the long-term co-evolution of microorganisms, or enhancing the disease resistance of plants by inducing the activity of plant defense enzyme systems. Tobacco occupies an important position in the field of traditional cash crop cultivation in China. How to combine biological control technology to provide a low-cost, highly efficient and environmentally friendly composite preparation for preventing and controlling tobacco fungal diseases and its use method is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0006] In view of this, the present invention provides a composite preparation for preventing and controlling tobacco fungal diseases, and the composite preparation includes the following components: quercetin, toosendanin, and a composite microbial preparation.

[0007] Further, the composite microbial preparation is composed of 150 - 460 million cfu / g of Bacillus subtilis, 40 - 150 million cfu / g of Rhizobium, and 170 - 350 million cfu / g of Bacillus amyloliquefaciens.

[0008] Further, the Bacillus subtilis comes from the China Center for Agricultural Culture Collection, and the preservation number is ACCC10118; the Rhizobium yanglingense GBXD - 30 comes from the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2021669; the Bacillus amyloliquefaciens comes from the China Center for Type Microorganism Collection, and the preservation number is CCTCC M2019877.

[0009] Further, the proportion of the composite microbial preparation in the composite bactericide is 1 - 8 wt%.

[0010] Preferably, the proportion of quercetin in the composite bactericide is 0.1 - 0.5 wt%, and the proportion of toosendanin is 0.3 - 0.8 wt%.

[0011] Further, the preparation method of the composite microbial preparation is as follows:

[0012] (1) Uniformly disperse montmorillonite in deionized water, and while stirring, add N - acetylglucosamine to modify montmorillonite to obtain modified montmorillonite. Then, successively perform filtration, washing with deionized water, drying, pulverization, and sieving to obtain modified montmorillonite powder;

[0013] (2) Disperse the modified montmorillonite powder in deionized water, then add the composite microorganisms, stir evenly, after sufficient adsorption, add sodium alginate, and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0014] (3) Drop the mixed solution into a calcium chloride solution for solidification, filter and dry to obtain a composite microbial agent.

[0015] Furthermore, in step (1), the temperature of the modification is 60 - 90 °C, the mass ratio of N-acetylglucosamine to montmorillonite is 1:3 - 8, the drying temperature is 55 - 65 °C, the drying time is 8 - 24 h, and the sieving refers to passing through a standard sieve of 150 - 200 mesh.

[0016] Furthermore, in step (2), the temperature is controlled within the range of 20 - 30 °C during adsorption.

[0017] Furthermore, in step (3), the concentration of the calcium chloride solution is 0.3 - 0.5 mol / L, the solidification time is 15 - 25 min, and the drying refers to drying in an oven at 65 °C to constant weight.

[0018] The present invention also provides a preparation method and a usage method of the composite agent for preventing and treating tobacco fungal diseases. The preparation method is: weigh each component in proportion, and after sufficient mixing, a composite agent for preventing and treating tobacco fungal diseases is obtained. The usage method is: mix the prepared composite agent with water to prepare a dispersion with a concentration of 0.5 - 1.5 wt%, and spray it on the roots of tobacco, and thus the prevention and treatment of tobacco fungal diseases can be achieved.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The raw materials used in the composite agent for preventing and treating tobacco fungal diseases provided by the present invention include quercetin, toosendanin, and a composite microbial agent. The composite agent has the characteristics of preventing and treating tobacco fungal diseases, slow release, long-acting drug delivery, cost reduction and efficiency increase, environmental protection, etc.

[0021] The present invention modifies montmorillonite with a modifier, enhancing the adsorption ability of montmorillonite to microorganisms. The present invention uses sodium alginate to coat the modified montmorillonite, which not only further strengthens the adsorption ability but also reduces the harm of the external environment to microorganisms, and helps to improve the colonization effect of microorganisms in the tobacco rhizosphere microenvironment.

[0022] The composite microbial agent adopted by the present invention is compounded from specific types of microorganisms, and they play a synergistic role with each other, further improving the colonization effect of the composite microorganisms in the tobacco rhizosphere microenvironment. Specific Embodiments

[0023] A composite preparation for preventing and controlling tobacco fungal diseases, the composite preparation comprising the following components: quercetin, toosendanin, and a composite microbial preparation.

[0024] In some embodiments of the present invention, the composite microbial preparation is composed of 150 - 460 million cfu / g of Bacillus subtilis, 40 - 150 million cfu / g of Rhizobium, and 170 - 350 million cfu / g of Bacillus amyloliquefaciens. Preferably, the composite microbial preparation is composed of 230 - 410 million cfu / g of Bacillus subtilis, 110 - 140 million cfu / g of Rhizobium, and 190 - 250 million cfu / g of Bacillus amyloliquefaciens. More preferably, the composite microbial preparation is composed of 350 million cfu / g of Bacillus subtilis, 120 million cfu / g of Rhizobium, and 230 million cfu / g of Bacillus amyloliquefaciens.

[0025] In some embodiments of the present invention, the Bacillus subtilis comes from the China Center for Agricultural Culture Collection, with the preservation number ACCC10118; the Rhizobium yanglingense GBXD - 30 comes from the China Center for Type Culture Collection, with the preservation number CCTCC NO: M2021669; the Bacillus amyloliquefaciens comes from the China Center for Type Microorganism Collection, with the preservation number CCTCC M2019877.

[0026] In some embodiments of the present invention, the proportion of the composite microbial preparation in the composite bactericide is 1 - 8 wt%, preferably, the proportion of the composite microbial preparation in the composite bactericide is 3 - 6 wt%, more preferably, the proportion of the composite microbial preparation in the composite bactericide is 5 wt%.

[0027] In some embodiments of the present invention, the proportion of quercetin in the composite bactericide is 0.1 - 0.5 wt%, preferably, the proportion of quercetin in the composite bactericide is 0.2 - 0.5 wt%, more preferably, the proportion of quercetin in the composite bactericide is 0.3 wt%.

[0028] In some embodiments of the present invention, the proportion of toosendanin in the composite bactericide is 0.3 - 0.8 wt%, preferably, the proportion of toosendanin in the composite bactericide is 0.4 - 0.6 wt%, more preferably, the proportion of toosendanin in the composite bactericide is 0.5 wt%.

[0029] The quercetin used in the present invention, also known as quercetin, quercetagetin, quercetin, with the CAS number 117 - 39 - 5, purity 95%, product model S33752, is purchased from Shanghai Yuanye Bio - technology Co., Ltd.

[0030] Toosendanin used in the present invention has a CAS number of 58812-37-6, a purity of ≥98%, and a product model of B20201. It is purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0031] Those skilled in the art know that microbial agents mainly control fungal diseases by means of the growth and reproduction of microorganisms to produce antibacterial substances or induce tobacco to produce resistance. However, this process is relatively slow to take effect compared to chemical pesticides. When fungal diseases break out suddenly and the disease spreads rapidly, applying traditional microbial agents may not be able to control the development of the disease in time. Therefore, in the present invention, a certain proportion of quercetin and toosendanin are added to the compound microbial agent. The results show that the compound microbial agent prepared by combining the three can quickly exert an antibacterial effect and effectively control tobacco fungal diseases. The reason may be that the addition of quercetin and toosendanin has the effect of stimulating the growth and development of microorganisms, promoting the rapid formation of dominant colonies by microorganisms at the roots of tobacco plants, competing for the growth space of pathogenic bacteria, and at the same time producing antibacterial substances, effectively inhibiting the occurrence of tobacco fungal diseases. And the inventors have found that the addition of quercetin and toosendanin can also promote the growth of tobacco roots and enhance the nutrient absorption ability of tobacco plants, thereby further improving the resistance of tobacco plants to fungal diseases.

[0032] In some embodiments of the present invention, the preparation method of the compound microbial agent is as follows:

[0033] (1) Montmorillonite is evenly dispersed in deionized water, and N-acetylglucosamine is added while stirring to modify montmorillonite to obtain modified montmorillonite. Then, filtration, washing with deionized water, drying, pulverizing, and sieving are carried out in sequence to obtain modified montmorillonite powder;

[0034] (2) The modified montmorillonite powder is dispersed in deionized water, and then compound microorganisms are added and stirred evenly. After sufficient adsorption, sodium alginate is added and stirred until sodium alginate is fully dissolved to obtain a mixed solution;

[0035] (3) The mixed solution is dropped into calcium chloride solution for curing, filtered, and dried to obtain the compound microbial agent.

[0036] In some embodiments of the present invention, the temperature of the modification in step (1) is 60-90°C. Preferably, the temperature of the modification is 65-80°C. More preferably, the temperature of the modification is 70°C.

[0037] In some embodiments of the present invention, the mass ratio of N-acetylglucosamine to montmorillonite is 1:3-8. Preferably, the mass ratio of N-acetylglucosamine to montmorillonite is 1:3-5. More preferably, the mass ratio of N-acetylglucosamine to montmorillonite is 1:3.5.

[0038] In some embodiments of the present invention, the drying temperature is 55 - 65°C, and the drying time is 8 - 24 h. Preferably, the drying temperature is 60 - 65°C, and the drying time is 12 - 24 h. More preferably, the drying temperature is 63°C, and the drying time is 18 h.

[0039] In some embodiments of the present invention, the sieving refers to sieving through a standard sieve of 150 - 200 mesh. Preferably, the sieving refers to sieving through a standard sieve of 150 mesh.

[0040] In some embodiments of the present invention, during the adsorption in step (2), the temperature is controlled within the range of 20 - 30°C. Preferably, the temperature during adsorption is controlled at 25°C.

[0041] In some embodiments of the present invention, the concentration of the calcium chloride solution in step (3) is 0.3 - 0.5 mol / L. Preferably, the concentration of the calcium chloride solution is 0.3 mol / L. The curing time in step (3) is 5 - 15 min. Preferably, the curing time is 8 min. The drying in step (3) refers to drying in an oven at 65°C until constant weight.

[0042] The present invention controls the curing time and the concentration of the calcium chloride solution within a reasonable range, which plays a role in ensuring that the external structure is loose and helps the release of microorganisms. In addition, those skilled in the art know that for beneficial microorganisms to exert their antibacterial effects, they first need to colonize in the rhizosphere of tobacco plants or at the site of pathogen infection. However, due to the influence of various factors in the soil environment, it is difficult for microorganisms to colonize in the soil. Therefore, the present invention uses modified montmorillonite and sodium alginate to encapsulate microorganisms, which can not only provide a relatively stable microenvironment for microorganisms, effectively isolate the interference of adverse factors in the environment such as temperature, humidity, light, oxygen, and harmful substances in the soil, play a role in protecting the microbial activity and improving the survival rate of microorganisms under different environmental conditions, but also, the modified montmorillonite can adsorb nutrients and moisture in the soil, which helps microorganisms to colonize and grow in the soil of tobacco roots.

[0043] The present invention can achieve the slow release of microorganisms by selecting appropriate materials and preparation processes, thereby improving the utilization rate of microbial agents, reducing the usage amount and frequency of use.

[0044] The present invention also provides a preparation method and a usage method for the composite agent for preventing and treating tobacco fungal diseases. The preparation method is: weighing each component according to the proportion and fully mixing them to obtain the composite agent for preventing and treating tobacco fungal diseases. The usage method is: mixing the prepared composite agent with water to prepare a dispersion with a concentration of 0.5 - 1.5 wt%, and spraying it on the roots of tobacco plants to achieve the prevention and treatment of tobacco fungal diseases.

[0045] To make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without any creative work fall within the scope of protection of the present invention.

[0046] In the following embodiments, the test methods or testing methods, unless otherwise specified, are all conventional methods; the raw materials and auxiliaries, unless otherwise specified, are all obtained from conventional commercial channels or prepared by conventional methods.

[0047] Example 1

[0048] A preparation method of a composite preparation for preventing and controlling tobacco fungal diseases, which consists of the following steps: mixing 0.3 wt% of quercetin, 0.5 wt% of toosendanin, 5 wt% of a composite microbial agent, and 94.2 wt% of water in proportion and dispersing them evenly to obtain a composite preparation for preventing and controlling tobacco fungal diseases.

[0049] The composite microbial agent consists of 350 million cfu / g of Bacillus subtilis, 120 million cfu / g of Rhizobium, and 230 million cfu / g of Bacillus amyloliquefaciens.

[0050] The preparation method of the composite microbial agent is as follows:

[0051] (1) Uniformly disperse 1.75 g of montmorillonite in 250 ml of deionized water, add 0.5 g of N-acetylglucosamine while stirring, modify the montmorillonite at a temperature of 70 °C, with the stirring speed of 600 r / min, to obtain modified montmorillonite. Then, filter, wash with deionized water three times, dry at a temperature of 63 °C for 18 h, and pulverize and pass through a 150-mesh standard sieve to obtain modified montmorillonite powder;

[0052] (2) Disperse the modified montmorillonite powder in 380 ml of deionized water, then add 0.5 g of composite microorganisms, stir evenly, control the environmental temperature at 25 °C for adsorption. After the adsorption is completed, add 5.0 g of sodium alginate and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0053] (3) Drop the mixed solution into a calcium chloride solution with a concentration of 0.3 mol / L, cure for 8 min, filter, and dry in an oven at 65 °C to constant weight to obtain a composite microbial agent.

[0054] Example 2

[0055] A preparation method of a composite preparation for preventing and controlling tobacco fungal diseases, which consists of the following steps: Mix 0.2 wt% of quercetin, 0.4 wt% of toosendanin, 2 wt% of a composite microbial preparation, and 97.4 wt% of water in proportion and disperse them evenly to obtain a composite preparation for preventing and controlling tobacco fungal diseases.

[0056] The composite microbial preparation consists of 150 million cfu / g of Bacillus subtilis, 40 million cfu / g of Rhizobium, and 170 million cfu / g of Bacillus amyloliquefaciens.

[0057] The preparation method of the composite microbial preparation is as follows:

[0058] (1) Uniformly disperse 2.4 g of montmorillonite in 300 ml of deionized water, add 0.8 g of N-acetylglucosamine while stirring, modify montmorillonite at a temperature of 60 °C, with the stirring speed of 600 r / min, to obtain modified montmorillonite. Then, successively carry out filtration, wash with deionized water 3 times, dry at a temperature of 55 °C for 8 h, and pulverize and pass through a 150-mesh standard sieve to obtain modified montmorillonite powder;

[0059] (2) Disperse the modified montmorillonite powder in 400 ml of deionized water, then add 0.5 g of composite microorganisms, stir evenly, control the environmental temperature at 20 °C for adsorption. After the adsorption is completed, add 5.0 g of sodium alginate and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0060] (3) Drop the mixed solution into a calcium chloride solution with a concentration of 0.3 mol / L, cure for 5 min, filter, and dry in an oven at 65 °C to constant weight to obtain a composite microbial preparation.

[0061] Example 3

[0062] A preparation method of a composite preparation for preventing and controlling tobacco fungal diseases, which consists of the following steps: Mix 0.3 wt% of quercetin, 0.5 wt% of toosendanin, 3 wt% of a composite microbial preparation, and 96.2 wt% of water in proportion and disperse them evenly to obtain a composite preparation for preventing and controlling tobacco fungal diseases.

[0063] The composite microbial preparation consists of 200 million cfu / g of Bacillus subtilis, 120 million cfu / g of Rhizobium, and 180 million cfu / g of Bacillus amyloliquefaciens.

[0064] The preparation method of the composite microbial preparation is as follows:

[0065] (1) Disperse 2.0 g of montmorillonite evenly in 350 ml of deionized water. While stirring, add 0.5 g of N - acetylglucosamine and modify the montmorillonite at a temperature of 70 °C. The stirring speed is 600 r / min to obtain modified montmorillonite. Then, successively carry out filtration, wash with deionized water three times, dry at a temperature of 58 °C for 12 h, pulverize and pass through a 150 - mesh standard sieve to obtain modified montmorillonite powder;

[0066] (2) Disperse the modified montmorillonite powder in 420 ml of deionized water. Then add 0.5 g of composite microorganisms, stir evenly, control the environmental temperature at 25 °C for adsorption. After adsorption is completed, add 5.0 g of sodium alginate and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0067] (3) Drop the mixed solution into a calcium chloride solution with a concentration of 0.4 mol / L, cure for 8 min, filter, and dry in an oven at 65 °C until constant weight to obtain a composite microbial agent.

[0068] Example 4

[0069] A preparation method of a composite agent for preventing and controlling tobacco fungal diseases, which consists of the following steps: Mix 0.5 wt% of quercetin, 0.5 wt% of toosendanin, 3.5 wt% of the composite microbial agent, and 95.5 wt% of water in proportion and disperse evenly to obtain a composite agent for preventing and controlling tobacco fungal diseases.

[0070] The composite microbial agent consists of 350 million cfu / g of Bacillus subtilis, 120 million cfu / g of Rhizobium, and 250 million cfu / g of Bacillus amyloliquefaciens.

[0071] The preparation method of the composite microbial agent is as follows:

[0072] (1) Disperse 3.5 g of montmorillonite evenly in 250 ml of deionized water. While stirring, add 0.5 g of N - acetylglucosamine and modify the montmorillonite at a temperature of 80 °C. The stirring speed is 600 r / min to obtain modified montmorillonite. Then, successively carry out filtration, wash with deionized water three times, dry at a temperature of 65 °C for 18 h, pulverize and pass through a 200 - mesh standard sieve to obtain modified montmorillonite powder;

[0073] (2) Disperse the modified montmorillonite powder in 450 ml of deionized water. Then add 0.5 g of composite microorganisms, stir evenly, control the environmental temperature at 25 °C for adsorption. After adsorption is completed, add 5.0 g of sodium alginate and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0074] (3) Drop the mixed solution into a calcium chloride solution with a concentration of 0.5 mol / L, solidify for 10 min, filter, and dry in an oven at 65 °C until constant weight to obtain the composite microbial agent.

[0075] Example 5

[0076] A preparation method of a composite agent for preventing and controlling tobacco fungal diseases, which consists of the following steps: Mix 0.5 wt% of quercetin, 0.6 wt% of toosendanin, 8 wt% of the composite microbial agent, and 90.9 wt% of water in proportion and disperse them evenly to obtain the composite agent for preventing and controlling tobacco fungal diseases.

[0077] The composite microbial agent consists of 460 million cfu / g of Bacillus subtilis, 150 million cfu / g of Rhizobium, and 350 million cfu / g of Bacillus amyloliquefaciens.

[0078] The preparation method of the composite microbial agent is as follows:

[0079] (1) Uniformly disperse 4.0 g of montmorillonite in 550 ml of deionized water, add 0.5 g of N-acetylglucosamine while stirring, modify the montmorillonite at a temperature of 90 °C, with the stirring speed of 600 r / min, to obtain modified montmorillonite. Then, filter, wash with deionized water 3 times, dry at a temperature of 65 °C for 24 h, and pulverize and pass through a 200-mesh standard sieve to obtain modified montmorillonite powder;

[0080] (2) Disperse the modified montmorillonite powder in 650 ml of deionized water, then add 0.5 g of the composite microorganism, stir evenly, control the environmental temperature at 30 °C for adsorption. After adsorption, add 5.0 g of sodium alginate and stir until the sodium alginate is fully dissolved to obtain a mixed solution;

[0081] (3) Drop the mixed solution into a calcium chloride solution with a concentration of 0.5 mol / L, solidify for 15 min, filter, and dry in an oven at 65 °C until constant weight to obtain the composite microbial agent.

[0082] Comparative Example 1

[0083] The difference from Example 1 is that quercetin and toosendanin are not added.

[0084] Comparative Example 2

[0085] The difference from Example 1 is that quercetin is not added.

[0086] Comparative Example 3

[0087] The difference from Example 1 is that toosendanin is not added.

[0088] Control effect test: The composite bacterial agent prepared in Examples 1-5 and Comparative Examples 1-3 was mixed with water to prepare a dispersion with a concentration of 0.5 wt%, and sprayed on the roots of tobacco at an application rate of 150 mL per plant.

[0089] The control effect of the composite bacterial agent prepared in Examples 1-5 and Comparative Examples 1-3 was tested, and the control effects of tobacco black shank, root rot, anthracnose, wilt, gray mold, white rot, brown spot disease, and virus diseases were statistically analyzed. The results are shown in Table 1.

[0090] Table 1

[0091]

[0092] As shown in Table 1, the combined use of quercetin and toosendanin has a positive effect on the prevention and treatment of tobacco fungal diseases, and can significantly improve the control effect of the composite microbial preparation on various tobacco fungal diseases. In addition, the control effect of the composite preparation with toosendanin added in Comparative Example 2 on tobacco fungal diseases is not as good as that of Comparative Example 1 and Examples 1-5, indicating that the addition of toosendanin alone may inhibit the growth of microorganisms, and when quercetin, toosendanin and the composite microbial preparation are used together, the three play a synergistic role, so that the control effect of the composite preparation on tobacco fungal diseases is significantly improved.

[0093] The present invention also tested the tobacco fungal disease prevention and control effect of the composite microorganisms coated with unmodified montmorillonite and sodium alginate, and the prevention and control effect was obviously inferior to that of the present invention.

[0094] In addition, the type of microorganism has an important impact on the prevention and control of fungal diseases. Specific types of microorganisms work synergistically with each other to improve the prevention and control of tobacco fungal diseases.

[0095] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composite preparation for preventing and controlling tobacco fungal diseases, characterized in that: The compound preparation comprises the following components: quercetin, toosendanin and compound microbial preparation.

2. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 1, characterized in that: The composite microbial preparation consists of 150-460 million cfu / g of Bacillus subtilis, 40-150 million cfu / g of Rhizobium, and 170-350 million cfu / g of Bacillus amyloliquefaciens.

3. The composite formulation for preventing and controlling tobacco fungal diseases according to claim 2, characterized in that: The Bacillus subtilis comes from the China Agricultural Microorganism Culture Collection Center with a collection number of ACCC10118; the Rhizobium comes from the China Center for Type Culture Collection with a collection number of CCTCCNO: M2021669; and the Bacillus amyloliquefaciens comes from the China Center for Type Microorganism Collection with a collection number of CCTCCM2019877.

4. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 1, characterized in that: The composite microbial preparation accounts for 1-8wt% of the composite bacterial agent.

5. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 1, characterized in that: The proportion of quercetin in the composite bacterial agent is 0.1-0.5wt%, and the proportion of toosendanin is 0.3-0.8wt%.

6. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 1, characterized in that: The preparation method of the composite microbial preparation is as follows: (1) uniformly dispersing montmorillonite in deionized water, adding N-acetylglucosamine while stirring to modify the montmorillonite to obtain modified montmorillonite, and then filtering, washing with deionized water, drying, crushing, and sieving in sequence to obtain modified montmorillonite powder; (2) dispersing the modified montmorillonite powder in deionized water, then adding the composite microorganisms, stirring evenly, and adding sodium alginate after sufficient adsorption, stirring until the sodium alginate is fully dissolved to obtain a mixed solution; (3) dropping the mixed solution into a calcium chloride solution, solidifying, filtering, and drying to obtain a composite microbial preparation.

7. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 6, characterized in that: The modification temperature in step (1) is 60-90° C., the mass ratio of N-acetylglucosamine to montmorillonite is 1:3-8, the drying temperature is 55-65° C., the drying time is 8-24 hours, and the sieving refers to sieving through a 150-200 mesh standard sieve.

8. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 6, characterized in that: During the adsorption in step (2), the temperature is controlled within the range of 20-30°C.

9. The composite preparation for preventing and controlling tobacco fungal diseases according to claim 6, characterized in that: The concentration of the calcium chloride solution in step (3) is 0.3-0.5 mol / L, the curing time is 15-25 min, and the drying refers to drying in an oven at 65° C. to constant weight.

10. The method for preparing the composite preparation for preventing and controlling tobacco fungal diseases according to claim 1: weigh each component in proportion, mix them thoroughly, and then obtain the composite preparation for preventing and controlling tobacco fungal diseases.

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