Composition containing sterol and amino-oligosaccharin as well as preparation and application of composition

Abstract

The invention discloses a composition containing sterol and amino-oligosaccharin, and belongs to the field of pesticides. The invention provides a composition containing sterol and amino-oligosaccharin, the active ingredients of the composition comprise the sterol and the amino-oligosaccharin, and the mass ratio of the sterol to the amino-oligosaccharin is (1: 25)-(1: 400). The composition provided by the invention shows a synergistic interaction effect on tobacco mosaic virus (TMV) within a specific ratio range.

Description

Technical Field

[0002] This invention belongs to the field of pesticides, specifically relating to a composition containing sterols and amino oligosaccharides, its preparation and application. Background Technology

[0004] Plant viral diseases have caused serious damage to agriculture in my country and are among the most difficult plant diseases to control in agricultural production worldwide. They mainly affect crops such as wheat, rice, tobacco, tomatoes, peppers, and potatoes, leading to problems such as declining quality, reduced yield, weakened resistance, and smaller or deformed fruits.

[0005] To control plant viral diseases, comprehensive measures are needed, including selecting virus-resistant varieties, seed and seedling treatment, field management, crop rotation, pesticide application, and controlling insect vectors. Among these, pesticide control is a crucial method. Currently, viral disease control in my country mainly relies on chemical pesticides such as morpholine guanidine hydrochloride. Long-term, single-use of morpholine guanidine hydrochloride can lead to increased plant virus resistance and weakened control efficacy. Furthermore, the extensive use of chemical pesticides results in pesticide residues, impacting food safety. Therefore, there is an urgent need to develop highly active, pollution-free, bio-based antiviral drugs, which are of great significance in green ecological agriculture.

[0006] Sterols are plant-derived sterol compounds that directly inhibit viral replication and induce host resistance, thereby indirectly preventing viral infection.

[0007] Amino oligosaccharides are natural polysaccharides derived from the deacetylation of chitin. They stimulate plant immunity (resistance inducers), inducing the production of disease-resistant proteins and phenolic substances, thereby enhancing systemic resistance. They are environmentally friendly, promote growth, and are less likely to induce pesticide resistance.

[0008] Sterenols and amino oligosaccharides, when used alone, have some effect in controlling crop viral diseases, but these diseases easily develop resistance. Through in-depth research, the inventors of this patent application have discovered that a specific ratio of sterols and amino oligosaccharides exhibits a synergistic effect in controlling plant viral diseases. Currently, there are no reports of combinations of sterols and amino oligosaccharides. Summary of the Invention

[0010] The purpose of this invention is to provide a composition containing sterols and amino oligosaccharides, which overcomes the drug resistance problem that is easily caused by single products, reduces the dosage, reduces the risk of drug residues, and improves the efficacy.

[0011] As is readily known to those skilled in the art, improving the efficacy of a composition and preventing the development of drug resistance often involves using multiple pesticides in combination or in combination. However, due to the lack of understanding of the components and their underlying mechanisms, it is difficult to predict whether the control effect of using multiple pesticides in combination or in combination is synergistic. Through in-depth experimental research, the inventors of this invention have discovered that the composition described in this invention exhibits a synergistic effect against tobacco mosaic virus (TMV) under specific formulations.

[0012] To achieve the technical objective of this invention, the invention is implemented through the following technical solution:

[0013] The present invention provides a composition containing sterol and amino oligosaccharide, wherein the active ingredients are composed of sterol and amino oligosaccharide, and the mass ratio of sterol and amino oligosaccharide is 1:25-1:400.

[0014] Furthermore, the present invention provides a composition containing sterol and amino oligosaccharide, wherein the mass ratio of sterol and amino oligosaccharide is 1:50-1:200.

[0015] Furthermore, the present invention provides a microemulsion containing the above composition, the preparation method of which is as follows: adding 5%-35% cyclohexanone, adding 0.01%-0.06% sterol, stirring to dissolve the sterol, adding 3%-6% calcium dodecylbenzenesulfonate, adding 8%-15% dodecyl alcohol polyoxyethylene polyoxypropylene ether, to prepare a sterol organic phase liquid; adding 40%-80% deionized water, adding 1%-5% amino oligosaccharide, stirring to dissolve the amino oligosaccharide, to prepare an amino oligosaccharide aqueous phase liquid; under continuous stirring of the sterol organic phase liquid, slowly adding the amino oligosaccharide aqueous phase liquid to the sterol organic phase liquid, finally preparing a transparent sterol-amino oligosaccharide microemulsion.

[0016] Furthermore, the present invention provides an application of the above composition in the prevention and control of tobacco mosaic virus disease (TMV).

[0017] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects:

[0018] 1. Reduced Risk of Resistance: In agricultural practice, long-term use of a single antiviral agent can easily lead to decreased efficacy. Sterols directly inhibit and indirectly induce resistance to plant viral diseases, while amino oligosaccharides indirectly induce resistance. Their mechanisms of action differ, and their combination reduces the risk of plant viruses developing resistance.

[0019] 2. Achieving synergistic effects: The combination of the two has a synergistic effect on tobacco mosaic virus (TMV), improving the control effect.

[0020] 3. In line with the trend of green agriculture: Both are plant-derived or natural derivatives, which are safer for agricultural products, non-target organisms and the environment compared with traditional chemical pesticides, and are in line with the direction of reducing pesticide use and green prevention and control. Detailed Implementation

[0022] The technical solution of the present invention will be described below with reference to embodiments. However, the present invention is not limited to the following embodiments. Unless otherwise specified, the experimental methods and detection methods described in each embodiment are conventional methods; unless otherwise specified, the reagents and materials can be purchased commercially.

[0023] Sterenol was purchased from Shaanxi Shangge Road Biotechnology Co., Ltd. Specification: 0.66%, Registration Certificate Number: PD20181614 .

[0024] Amino oligosaccharides were purchased from Shandong Jingbo Agricultural Chemical Technology Co., Ltd. Specifications: 85%, Registration Certificate Number: PD20240014 .

[0025] Formulation Examples

[0026] Example 1: 1.3% sterol-amino oligosaccharide microemulsion (1:25)

[0027] Add 280 kg of cyclohexanone and 75.8 kg of 0.66% sterol precursor, stir to dissolve the sterol, add 45 kg of calcium dodecylbenzenesulfonate and 135 kg of dodecyl alcohol polyoxyethylene polyoxypropylene ether to prepare a sterol organic phase; add 449.5 kg of deionized water and 14.7 kg of 85% amino oligosaccharide precursor, stir to dissolve the amino oligosaccharide to prepare an amino oligosaccharide aqueous phase; while continuously stirring the sterol organic phase, slowly add the amino oligosaccharide aqueous phase to the sterol organic phase to finally prepare a transparent 1.3% sterol·amino oligosaccharide microemulsion.

[0028] Example 2: 2.04% sterol-amino oligosaccharide soluble concentrate (1:50)

[0029] Add 250 kg of cyclohexanone and 60.7 kg of 0.66% sterol precursor, stir to dissolve the sterol, add 45 kg of calcium dodecylbenzenesulfonate and 135 kg of dodecyl alcohol polyoxyethylene polyoxypropylene ether to prepare a sterol organic phase; add 485.7 kg of deionized water and 23.6 kg of 85% amino oligosaccharide precursor, stir to dissolve the amino oligosaccharide to prepare an amino oligosaccharide aqueous phase; while continuously stirring the sterol organic phase, slowly add the amino oligosaccharide aqueous phase to the sterol organic phase to finally prepare a transparent 2.04% sterol·amino oligosaccharide microemulsion.

[0030] Example 3: 1.01% Sterenol·Amino Oligosaccharide Soluble Solution (1:100)

[0031] Add 70 kg of cyclohexanone and 15.2 kg of 0.66% sterol precursor, stir to dissolve the sterol, add 35 kg of calcium dodecylbenzenesulfonate and 105 kg of dodecyl alcohol polyoxyethylene polyoxypropylene ether to prepare a sterol organic phase; add 763 kg of deionized water and 11.8 kg of 85% amino oligosaccharide precursor, stir to dissolve the amino oligosaccharide to prepare an amino oligosaccharide aqueous phase; while continuously stirring the sterol organic phase, slowly add the amino oligosaccharide aqueous phase to the sterol organic phase to finally prepare a transparent 1.01% sterol·amino oligosaccharide microemulsion.

[0032] Example 4: 4.02% Sterenol·Amino Oligosaccharide Soluble Solution (1:200)

[0033] Add 150 kg of cyclohexanone and 30.4 kg of 0.66% sterol precursor, stir to dissolve the sterol, add 40 kg of calcium dodecylbenzenesulfonate and 120 kg of dodecyl alcohol polyoxyethylene polyoxypropylene ether to prepare a sterol organic phase; add 612.5 kg of deionized water and 47.1 kg of 85% amino oligosaccharide precursor, stir to dissolve the amino oligosaccharide to prepare an amino oligosaccharide aqueous phase; while continuously stirring the sterol organic phase, slowly add the amino oligosaccharide aqueous phase to the sterol organic phase to finally prepare a transparent 4.02% sterol·amino oligosaccharide microemulsion.

[0034] Example 5: 4.01% Sterenol·Amino Oligosaccharide Soluble Solution (1:400)

[0035] Add 70 kg of cyclohexanone and 15.2 kg of 0.66% sterol precursor, stir to dissolve the sterol, add 35 kg of calcium dodecylbenzenesulfonate and 105 kg of dodecyl alcohol polyoxyethylene polyoxypropylene ether to prepare a sterol organic phase; add 727.7 kg of deionized water and 47.1 kg of 85% amino oligosaccharide precursor, stir to dissolve the amino oligosaccharide to prepare an amino oligosaccharide aqueous phase; under continuous stirring of the sterol organic phase, slowly add the amino oligosaccharide aqueous phase to the sterol organic phase to finally prepare a transparent 4.01% sterol·amino oligosaccharide microemulsion.

[0036] Biometrics Example

[0037] Reagent preparation: Dissolve the test reagent sterol in cyclohexanone, dissolve the amino oligosaccharide in water, and then dilute with 0.5% Tween-80 aqueous solution to prepare single-agent stock solutions and stock solutions of each proportioned mixture. Set up 5 mass concentration gradients for both single-agent and each proportioned mixture according to the same ratio method for later use.

[0038] Target of control: Tobacco mosaic virus (TMV)

[0039] Preparation of inoculated virus solution: Select upper tobacco leaves that have been inoculated with TMV for more than 3 weeks, homogenize them in phosphate buffer, filter them with double gauze, centrifuge at 8000 r / min, treat the supernatant twice with polyethylene glycol, centrifuge again, and suspend the precipitate in phosphate buffer to obtain the TMV extract. The entire experiment was carried out at 4℃.

[0040] Experimental crop: Select tobacco plants with good growth and uniform leaf size.

[0041] Experimental Method: A spraying method was used. The test solution was sprayed onto the tobacco plants using a small sprayer until the entire leaf was wetted. Each treatment was repeated three times, with a water control included. After 24 hours, the tobacco leaves were sprinkled with carborundum (emery), and then brushed with a 500-fold diluted virus solution using a brush. After 30 minutes, the carborundum was rinsed off, and the leaves were placed in an artificial climate chamber for 2-3 days. The spots on the tobacco leaves were counted, and the inhibition rate was calculated.

[0042] Regression analysis was performed based on the logarithmic values ​​of each drug concentration and the corresponding inhibition rate probabilities to calculate the EC50 of each drug. 50 The interaction of the mixtures was evaluated using the Wadley method, where SR = 0.5-1.5 indicates an additive effect between the two agents; SR ≤ 0.5 indicates an antagonistic effect; and SR ≥ 1.5 indicates a synergistic effect. The combined effects of sterols and amino oligosaccharides on tobacco mosaic virus (TMV) are shown in Table 1.

[0043] EC 50 (Theoretical value) = (a+b) / (a / EC) 50 a+b / EC 50 b);

[0044] SR=EC 50 (Theoretical value) / EC 50 (Actual value);

[0045] Where SR is the synergistic coefficient, and a and b are the percentage contents of each component in the mixture.

[0046] Table 1. Results of the combined effects of sterols and amino oligosaccharides on tobacco mosaic virus (TMV).

[0047] Drug Name and Proportion <![CDATA[EC 50 Actual value (mg / L) <![CDATA[EC 50 Theoretical value (mg / L) SR Sterenol 6.26 / / Amino oligosaccharides 19.74 / / Sterenol: Amino oligosaccharide = 1:5 9.95 14.53 1.46 Sterenol: Amino oligosaccharide = 1:25 8.48 18.23 2.15 Sterenol: Amino oligosaccharide = 1:50 5.70 18.94 3.32 Sterenol: Amino oligosaccharide = 1:100 4.77 19.33 4.05 Sterenol: Amino oligosaccharide = 1:200 6.16 19.53 3.17 Sterenol: Amino oligosaccharide = 1:400 10.02 19.63 1.96 Sterenol: Amino oligosaccharide = 1:600 14.25 19.67 1.38

[0048] Combined crop assays showed that sterols and amino oligosaccharides, when combined in specific ratios, exhibited good control efficacy against tobacco mosaic virus (TMV). When sterols and amino oligosaccharides were combined in a ratio of 1:25–1:400, the synergistic effect was greater than 1.5, demonstrating a significant synergistic effect. In particular, when the ratios were combined in a ratio of 1:50–1:200, the synergistic effect was greater than 3.0, indicating a substantial synergistic effect.

[0049] As described above, the present invention can be well implemented. The above embodiments are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, all kinds of changes and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the present invention.

Claims

1. A composition containing a sterol and an aminosaccharide, the effective components of which consist of the sterol and the aminosaccharide, the mass ratio of the sterol and the aminosaccharide being 1:25-1:

400. The agricultural composition of the present application is characterized in that it contains a sterol registered under the registration number: PD20181614 , and an amino-oligosaccharide registered under the registration number: PD20240014 .

2. The agricultural composition of claim 1, wherein: The mass ratio of the sterol and the aminosaccharide is 1:50-1:

200.

3. A microemulsion comprising the composition of claim 1, characterized in that: The preparation method is as follows: adding cyclohexanone 5%-35%, adding sterol 0.01%-0.06%, stirring to dissolve the sterol, adding calcium dodecylbenzenesulfonate 3%-6%, adding dodecanol polyoxyethylene polyoxypropylene ether 8%-15%, to prepare a sterol organic phase liquid; adding water 40%-80%, adding aminosaccharide 1%-5%, stirring to dissolve the aminosaccharide, to prepare an aminosaccharide aqueous phase liquid; under continuous stirring of the sterol organic phase liquid, slowly adding the aminosaccharide aqueous phase liquid into the sterol organic phase liquid, to finally prepare a transparent sterol-aminosaccharide microemulsion.

4. Use of a composition according to claim 1, characterized in that: For the prevention and treatment of tobacco mosaic virus (TMV).