Preparation for preventing and treating plant virus disease and application thereof

Abstract

The application discloses a preparation for preventing and treating plant viral diseases and application thereof, and belongs to the technical field of plant disease prevention and treatment. The preparation for preventing and treating plant viral diseases has at least one of 4-vinyl-2,6-dimethoxyphenol, (3beta, 24R)-ergosta-5-en-3-ol, (3beta, 22E)-stigmasta-5,22-dien-3-ol and (3beta)-stigmasta-5-en-3-ol as an effective component, and the content of the effective component is 0.625-20 mg / mL. The active component in the preparation has a good prevention and treatment effect on tobacco mosaic virus disease and pepper vein banding virus disease, wherein the combination of 4-vinyl-2,6-dimethoxyphenol and (3beta, 22E)-stigmasta-5,22-dien-3-ol has a prevention and treatment effect on tobacco mosaic virus disease and pepper vein banding virus disease greater than that of a single component, and has an important application prospect in the prevention and treatment of plant viral diseases.

Description

Technical Field

[0001] This invention belongs to the field of plant disease control technology, specifically relating to an agent for the prevention and control of plant viral diseases and its application. Background Technology

[0002] Plant viral diseases are among the major threats to crops, often referred to as "plant cancer." Tobacco mosaic virus (TMV) and chili veinal mottle virus (ChiVMV) are two particularly representative plant viruses that can infect various economic crops such as tobacco, chili peppers, and tomatoes, causing mosaic patterns, blisters, yellowing, wrinkling, necrosis, and stunted growth, severely impacting crop yield and quality. Currently, there are no specific treatments for plant viral diseases. Production mainly adopts a "prevention-oriented, integrated control" strategy, including the breeding of resistant varieties, agricultural measures, and chemical control. However, resistant varieties are limited and easily lose their resistance due to viral mutations; agricultural measures are inconsistent in effectiveness; and conventional chemical agents have weak direct inhibitory effects on the viruses themselves, making it difficult to fundamentally control the occurrence and spread of viral diseases.

[0003] In recent years, plant-derived bioactive substances have become a hot topic in plant viral disease control research due to their environmental friendliness, unique mechanisms of action, and low likelihood of developing resistance. Therefore, developing a highly efficient, broad-spectrum, and stable plant-derived antiviral agent has significant application value and practical implications. Summary of the Invention

[0004] In view of the problems existing in the prior art, the purpose of this invention is to provide an agent for preventing and controlling plant viral diseases and its application, so as to solve the technical problems such as the unsatisfactory effect of antiviral agents and the insufficient activity of single components in the prior art.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: An agent for the prevention and control of plant viral diseases, wherein the active ingredient is at least one of 4-vinyl-2,6-dimethoxyphenol, (3β,24R)-ergoster-5-en-3-ol, (3β,22E)-stigmaster-5,22-dien-3-ol, and (3β)-stigmaster-5-en-3-ol, and the content of the active ingredient is 0.625~20 mg / mL.

[0006] Based on the above scheme, the active ingredients are 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol.

[0007] Based on the above scheme, the concentration ratio of 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol is 1:1 to 1:8.

[0008] Based on the above scheme, it further includes pesticide-acceptable solvents or adjuvants.

[0009] The application of the above-mentioned agents for controlling plant viral diseases.

[0010] Based on the above scheme, the plant viral diseases mentioned are plant diseases caused by tobacco mosaic virus or pepper vein mottle virus.

[0011] Based on the above scheme, the plant mentioned is tobacco.

[0012] Based on the above plan, the above-mentioned agents for preventing and controlling plant viral diseases are sprayed onto the plant leaves to achieve the purpose of preventing and controlling plant viral diseases.

[0013] Advantages of the technical solution of this invention This invention provides a formulation for controlling plant viral diseases. The active ingredient in this formulation is a plant-derived compound that is easy to extract, highly active, and possesses the characteristics of being green and economical. It also exhibits good control effects against tobacco mosaic virus and pepper vein mottle virus. Specifically, the combination of 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol demonstrates a greater control effect against tobacco mosaic virus and pepper vein mottle virus than a single component. The two components have a synergistic effect, which can compensate for the antiviral effect achieved by higher concentrations of a single component. This formulation has significant application prospects in the control of plant viral diseases. Attached Figure Description

[0014] Figure 1 The diagram shows the control effect of the combined solution on tobacco mosaic virus and pepper vein mottle virus. Detailed Implementation

[0015] The terminology used in this invention, unless otherwise specified, generally has the meanings commonly understood by those skilled in the art. The invention is further described in detail below with reference to specific embodiments and data. The following embodiments are merely illustrative and are not intended to limit the scope of the invention in any way.

[0016] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the experimental materials, reagents, and chemicals used in the following embodiments can be obtained through general channels.

[0017] In the following examples, 4-vinyl-2,6-dimethoxyphenol (rapeseed polyphenol, CAS No. 28343-22-8), (3β,24R)-ergoster-5-en-3-ol (rapeseed sterol CAS No.: 474-62-4), (3β,22E)-stigmaster-5,22-dien-3-ol (stigmasterol, CAS No. 83-48-7), and (3β)-stigmaster-5-en-3-ol (β-sitosterol CAS No.: 83-46-5) were purchased from Shanghai Yuanye Biotechnology Co., Ltd., and all had a purity of ≥95% by HPLC.

[0018] The tobacco varieties Sansheng NN and Zhongyan 100 were derived from germplasm resources of the Tobacco Research Institute of the Chinese Academy of Agricultural Sciences, while the plant viruses TMV and ChiVMV were preserved by the Virus Research Group of the Tobacco Research Institute of the Chinese Academy of Agricultural Sciences.

[0019] Example 1

[0020] A compound preparation for preventing and treating tobacco viral diseases, wherein the active ingredient is any one of 4-vinyl-2,6-dimethoxyphenol, (3β,24R)-ergoster-5-en-3-ol, (3β,22E)-stigmaster-5,22-dien-3-ol, and (3β)-stigmaster-5-en-3-ol, and the content of the active ingredient is 0.625~20 mg / mL.

[0021] 4-Vinyl-2,6-dimethoxyphenol, (3β,24R)-ergoster-5-en-3-ol, (3β,22E)-stigmaster-5,22-dien-3-ol, and (3β)-stigmaster-5-en-3-ol were dissolved in small amounts of anhydrous ethanol or dimethyl sulfoxide, and then prepared into 20 mg / mL solutions with phosphate-buffered saline (PBS). The inhibition rate of different compound solutions on TMV was detected by the plaque assay. The specific method is as follows: Different compound solutions were mixed with an equal volume of TMV extract (TMV extract was obtained by grinding 1g of diseased leaves, adding 60mL of phosphate buffer, homogenizing, and filtering through lens paper, the same below) to form treatment groups, and PBS mixed with an equal volume of TMV extract to form a control group. 30-45 minutes after mixing, the treatment solution was rubbed onto one side of a leaf of the NN plant (a host plant with 6-7 leaves) at the necrotic stage, and the control solution was rubbed onto the other side. Each treatment consisted of 5 leaves, with 4 replicates. Three days after inoculation, the number of necrotic spots was recorded using a counter, and the inhibition rate against TMV was calculated.

[0022] Table 1. Inhibitory effects of different compounds on TMV

[0023] The inhibitory effects of different compounds on TMV are shown in Table 1. At a concentration of 20 mg / mL, all four compounds showed significant inhibitory effects on TMV. Among them, 4-vinyl-2,6-dimethoxyphenol showed the best effect, with an inhibition rate of 93.52%. (3β,24R)-ergoster-5-en-3-ol, (3β,22E)-stigmaster-5,22-dien-3-ol and (3β)-stigmaster-5-en-3-ol also showed certain inhibitory effects on TMV, with inhibition rates of 50.72%, 52.06% and 51.23%, respectively, and there was no significant difference among the three.

[0024] Example 2

[0025] A compound preparation for preventing and treating tobacco viral diseases, the active ingredients being 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol; wherein the concentration of 4-vinyl-2,6-dimethoxyphenol is 0.625~20 mg / mL, the concentration of (3β,22E)-stigmaster-5,22-dien-3-ol is 0.625~20 mg / mL, and the concentration ratio of 4-vinyl-2,6-dimethoxyphenol to (3β,22E)-stigmaster-5,22-dien-3-ol is 1:1~1:8.

[0026] Inhibitory effect of compound preparations for the prevention and treatment of tobacco viral diseases on TMV Prepare the following solutions respectively: 4-Vinyl-2,6-dimethoxyphenol 0.625, 1.25, 2.5, 5, 10, 20 mg / mL; (3β,22E)-stigmaster-5,22-dien-3-ol 0.625, 1.25, 2.5, 5, 10, 20 mg / mL; 4-Vinyl-2,6-dimethoxyphenol + (3β,22E)-stigmaster-5,22-dien-3-ol 2.5+2.5 mg / mL, 2.5+5 mg / mL, 2.5+10 mg / mL, 2.5+20 mg / mL.

[0027] The compounds were first dissolved in a small amount of anhydrous ethanol or dimethyl sulfoxide, and then different compound formulation solutions were prepared with phosphate-buffered saline (PBS).

[0028] The inhibition rate of different formulations against TMV was determined using the necrotic spot method. The specific method is as follows: Different compound formulations were mixed with an equal volume of TMV extract to form treatment groups, while a mixture of PBS and an equal volume of TMV extract served as a control group. 30-45 minutes after mixing, the treatment solution was rubbed onto one side of a leaf of the 6-7 leaf stage host plant (Trifolium NN) with necrotic spots, and the control solution was inoculated onto the other side. Each treatment consisted of 5 leaves, with 4 replicates. Three days after inoculation, the number of necrotic spots was recorded using a counter, and the inhibition rate against TMV was calculated.

[0029] Table 2. Inhibitory effects of different compound formulations on TMV

[0030] The experimental results are shown in Table 2: the inhibitory effect of different concentrations of compounds on TMV decreased significantly with decreasing concentration. Among them, 4-vinyl-2,6-dimethoxyphenol showed an inhibition rate of 50.33% at 2.5 mg / mL; (3β,22E)-stigmaster-5,22-dien-3-ol showed an inhibition rate of 52.29% at 20 mg / mL; the combination of 4-vinyl-2,6-dimethoxyphenol + (3β,22E)-stigmaster-5,22-dien-3-ol showed a significantly higher inhibitory effect on TMV than the single compound, with an inhibition rate of 93.56% at 2.5 ± 5 mg / mL.

[0031] Application of compound preparations in the prevention and treatment of tobacco viral diseases To prepare a compositional solution containing 2.5 mg / mL of 4-vinyl-2,6-dimethoxyphenol and 5 mg / mL of (3β,22E)-stigmaster-5,22-dien-3-ol, the compounds were first dissolved in a small amount of anhydrous ethanol or dimethyl sulfoxide, and then prepared into a compositional solution using phosphate-buffered saline (PBS).

[0032] For tobacco plants at the 6-7 leaf stage, the above-mentioned composition solution was sprayed on the leaves of Zhongyan 100, with three consecutive sprays at 3-5 day intervals. Water spraying served as a blank control, and a 750-fold dilution of 0.06% sterol microemulsion served as a control. Each treatment consisted of 15 plants, replicated four times. 15-30 minutes after the last spray, TMV or ChiVMV disease sap was rubbed inoculated (ChiVMV disease sap was obtained by grinding 1g of diseased leaves, adding 20mL of phosphate buffer, homogenizing, and filtering through lens paper). 14-21 days after inoculation, the disease severity level of all tobacco plants in each treatment was investigated, and the control effect was calculated. Disease severity levels were determined on a per-plant basis, according to the criteria shown in Table 3.

[0033] Table 3. Criteria for Determining the Severity Levels of Tobacco Mosaic Virus and Pepper Vein Mottle Virus

[0034] Note: The range of 1 / 3 to 1 / 2 includes 1 / 3. The rest are the same.

[0035] The incidence rate, disease index, and prevention effect are calculated using the following formulas:

[0036] The test results are as follows Figure 1 As shown in Table 4.

[0037] Table 4. Control efficacy of the combined solution against tobacco mosaic virus and pepper vein mottle virus.

[0038] Figure 1 The efficacy of the composition solution against tobacco mosaic virus (TMV) was demonstrated. As shown in the figure, 14 days after TMV inoculation, the mosaic and blister symptoms in the treatment group sprayed with the antiviral composition and the control group sprayed with 0.06% sterol microemulsion were significantly milder than those in the blank control group sprayed with water. 21 days after ChiVMV inoculation, compared with the water-sprayed control, both the antiviral composition and the 0.06% sterol microemulsion significantly inhibited the spread of yellow spot and mottling symptoms.

[0039] Table 4 shows the control efficacy of the composition solution against tobacco mosaic virus (TMV) and pepper vein mottle virus (PMV). As shown in Table 4, 14 days after TMV inoculation, the disease index of the composition-treated group was 17.36, which was not significantly different from the control group's (17.78); both were significantly lower than the water control's (36.67). The antiviral composition achieved a preventive effect of 52.66% against TMV. 21 days after ChiVMV inoculation, the disease index of the antiviral composition-treated group was 17.04, and the control group's was 20.93; both were significantly lower than the water control's (44.81). The antiviral composition achieved a preventive effect of 61.97% against ChiVMV, slightly higher than the control group's (53.29%).

[0040] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A preparation for preventing and controlling plant viral diseases, characterized in that, The active ingredients are 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol, wherein the concentration ratio of 4-vinyl-2,6-dimethoxyphenol and (3β,22E)-stigmaster-5,22-dien-3-ol is 1:1 to 1:8, and the content of the active ingredients is 2.5 to 20 mg / mL.

2. The preparation for preventing and controlling plant viral diseases according to claim 1, characterized in that, It may further include pesticide-acceptable solvents or adjuvants.

3. The application of the preparation for controlling plant viral diseases according to any one of claims 1 to 2 in the control of plant viral diseases, characterized in that, The plant viral diseases mentioned are plant diseases caused by tobacco mosaic virus or pepper vein mottle virus; the plant mentioned is tobacco.

4. The application of the preparation for controlling plant viral diseases according to claim 3 in the control of plant viral diseases, characterized in that, The preparation for preventing and controlling plant viral diseases according to any one of claims 1 to 2 is sprayed onto the leaves of plants to achieve the purpose of preventing and controlling plant viral diseases.

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