阅读说明：本技术 小茴香提取物制备抗植物病毒剂的应用及抗植物病毒剂 (Application of fennel extract in preparation of anti-plant virus agent and anti-plant virus agent ) 是由 闫合 郭文慧 何军 周一万 吴华 冯俊涛 于 2020-05-06 设计创作，主要内容包括：本发明公开了小茴香提取物制备抗植物病毒剂的应用及抗植物病毒剂。小茴香提取物用于制备抗植物病毒剂的应用；所述的小茴香提取物为：小茴香经有机溶剂浸提后得到的浓缩浸膏。用于烟草、黄瓜、马铃薯等作物上防治烟草花叶病毒病(Tobacco mosaic virus,TMV)、黄瓜花叶病毒病(Cucumber mosaic virus,CMV)及马铃薯病毒病(Potato Virus X,PVX；Potato Virus Y,PVY)等多种植物病毒病。本发明的抗病毒剂抗病毒效果好,制作方式简单,成本低廉,具有开发为植物源抗病毒剂的潜力。(The invention discloses application of fennel extract in preparation of an anti-plant virus agent and the anti-plant virus agent. Use of fennel extract for the preparation of an anti-plant virus agent; the fennel extract is as follows: extracting fennel with organic solvent to obtain concentrated extract. The composition is used for preventing and treating various plant Virus diseases such as Tobacco Mosaic Virus (TMV), Cucumber Mosaic Virus (CMV) and Potato Virus (PVX) on crops such as Tobacco, Cucumber, Potato and the like. The antiviral agent has good antiviral effect, simple preparation method and low cost, and has the potential of being developed into a plant source antiviral agent.)

1. Use of fennel extract for the preparation of an anti-plant virus agent;

the fennel extract is as follows: extracting fennel with organic solvent to obtain concentrated extract.

The content of the fennel extract in the plant virus resisting agent is 10-60% by mass.

2. The use according to claim 1, wherein the fennel extract is: adding an extraction solvent into fennel powder with the particle size of 1-5 mm according to a ratio of 1:10(w/v), wherein the extraction solvent is any one or a mixture of more than two of methanol, ethanol, ethyl acetate, chloroform and petroleum ether, and standing at room temperature for 24 hours;

performing ultrasonic extraction at 40 deg.C and frequency of 100Hz for 60min, filtering, extracting the residue for 2 times, mixing the filtrates, and concentrating under reduced pressure to obtain plant extract.

3. The use according to claim 1 or 2, wherein the anti-plant virus agent is a soluble powder prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, stabilizer: 0-5%, surfactant: 5 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

4. The use of claim 1 or 2, wherein the plant virus resisting agent is water dispersible granules prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, surfactant: 7 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

5. The use according to claim 1 or 2, wherein the anti-plant virus agent is a soluble liquid agent, and is prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -40%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

6. The use of claim 1 or 2, wherein the plant virus resisting agent is an aqueous emulsion prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -30%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

7. An anti-plant virus agent is characterized in that fennel extract is used as a medicinal component, and the content of the fennel extract in percentage by mass is 10% -60%;

the fennel extract is as follows: extracting fennel with organic solvent to obtain concentrated extract.

8. The plant virus resisting agent is characterized in that the plant virus resisting agent is soluble powder and is prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, stabilizer: 0-5%, surfactant: 5-20 percent, and the sum of the weight percentages of the raw materials is 100 percent;

or the plant virus resisting agent is water dispersible granules prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, surfactant: 7 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

9. The plant virus resisting agent is characterized by being a soluble liquid agent and prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -40%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water, and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%;

or the plant virus resisting agent is an aqueous emulsion prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -30%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

10. The anti-plant virus agent according to any one of claims 7 to 9, wherein the fennel extract is: adding an extraction solvent into fennel powder with the particle size of 1-5 mm according to a ratio of 1:10(w/v), wherein the extraction solvent is any one or a mixture of more than two of methanol, ethanol, ethyl acetate, chloroform and petroleum ether, and standing at room temperature for 24 hours;

performing ultrasonic extraction at 40 deg.C and frequency of 100Hz for 60min, filtering, extracting the residue for 2 times, mixing the filtrates, and concentrating under reduced pressure to obtain plant extract.

Technical Field

The invention belongs to the field of plants and application thereof, and particularly relates to application of fennel extract in preparation of an anti-plant virus agent and the anti-plant virus agent.

Background

Plant viral diseases are important diseases in crop production, with economic losses due to viral diseases amounting to about four billion dollars each year worldwide. Several common plant viruses such as Tobacco Mosaic Virus (TMV), Cucumber Mosaic Virus (CMV), potato virus (PVX, PVY), etc. can infect over 800 crops, causing significant losses. Because viruses are replicated and amplified in plants by depending on host nutritional substrates, the commonly used medicaments for preventing and treating the viruses at present are difficult to directly act on virus particles so as to achieve ideal prevention and treatment effects.

Chemical pesticides still play no alternative role in disease control, but face the pressure of safety, environment, ecology and the like. At present, methods such as antiviral breeding, cross protection of attenuated strains, improvement of cultivation technology, control of virus transmission media and transgenosis by pesticides, and the like are all included, and the control of viral diseases by using natural products is a research hotspot in recent years. The plant extract contains various natural products with biological activity, generally has various and novel chemical structures, unique biological activity mechanism and capability of improving crop resistance; and the pesticide composition has the characteristics of good environmental compatibility, delaying the generation of drug resistance, promoting the healthy growth of crops and the like, and is an important resource for developing medicines and pesticides.

Fennel (Foeniculum vulgare) is a perennial herb of Umbelliferae, is distributed in most regions of the country, and the fruit, stem and leaf parts of the fennel can be used as medicines. The main metabolites in fennel comprise various compounds such as fatty oil, volatile oil, sterol, glucoside, amino acid, triterpene, tannin, flavone, cardiac glycoside, alkaloid, saponin, coumarin, volatile alkali, anthraquinone, etc. Pharmacological studies show that the fennel has the effects of dispelling cold, relieving pain, invigorating stomach, dispelling wind, clearing heat, reducing phlegm and the like. The literature reports that volatile oil in fennel has good antibacterial activity, and flavonoids, phenols, saponins, tannins, alkaloids and polysaccharides have antioxidant capacity. In addition, the fennel crude extract has a killing effect on the pine wood nematodes and also has an inhibition effect on bacteria carried by the pine wood nematodes. However, the research of fennel on the aspect of resisting plant viruses has not been reported, and the application of fennel on agricultural pest control needs to be further developed.

The efficient plant source natural product has the characteristics of safety, lasting effect, slow resistance development and the like. Some natural products can enhance the resistance of plants to diseases and insect pests and reduce economic loss by exciting the defense mechanism of plant bodies. Therefore, the application of the natural product can effectively reduce the application of chemical pesticides, thereby reducing a series of negative effects caused by the pesticides and having good ecological significance. Therefore, the application of the plant source natural product antiviral agent is a plant protection method which is good in ecological environment compatibility in plants. The research on the disease resistance of natural products has very important significance in the aspects of agricultural sustainable development, ecological environment protection, grain and food safety and the like.

Disclosure of Invention

The first purpose of the invention is to use fennel extract as a raw material for preparing an anti-plant virus agent;

the second purpose of the invention is to research the anti-plant virus agent with antiviral activity by using fennel extract as a raw material.

In order to achieve the purpose, the technical scheme adopted by the invention comprises the following steps:

according to a first object of the present invention, the use of an extract of fennel for the preparation of an anti-plant virus agent;

the fennel extract is as follows: extracting fennel with organic solvent to obtain concentrated extract.

The content of the fennel extract in the plant virus resisting agent is 10-60% by mass.

Optionally, the fennel extract is: adding an extraction solvent into fennel powder with the particle size of 1-5 mm according to a ratio of 1:10(w/v), wherein the extraction solvent is any one or a mixture of more than two of methanol, ethanol, ethyl acetate, chloroform and petroleum ether, and standing at room temperature for 24 hours;

performing ultrasonic extraction at 40 deg.C and frequency of 100Hz for 60min, filtering, extracting the residue for 2 times, mixing the filtrates, and concentrating under reduced pressure to obtain plant extract.

Optionally, the plant virus resisting agent is soluble powder prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, stabilizer: 0-5%, surfactant: 5 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

Optionally, the plant virus resisting agent is water dispersible granules prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, surfactant: 7 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

Optionally, the plant virus resisting agent is a soluble liquid agent, and is prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -40%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

Optionally, the plant virus resisting agent is an aqueous emulsion prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -30%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

An anti-plant virus agent, which takes fennel extract as medicinal component, the mass percentage content of the fennel extract is 10-60%;

the fennel extract is as follows: extracting fennel with organic solvent to obtain concentrated extract.

The plant virus resisting agent is soluble powder and is prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, stabilizer: 0-5%, surfactant: 5-20 percent, and the sum of the weight percentages of the raw materials is 100 percent;

or the plant virus resisting agent is water dispersible granules prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, and filler: 10% -60%, dispersant: 0-10%, surfactant: 7 to 20 percent, and the sum of the weight percentages of the raw materials is 100 percent.

The plant virus resisting agent is a soluble liquid and is prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -40%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water, and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%;

or the plant virus resisting agent is an aqueous emulsion prepared from the following raw materials in percentage by weight:

10-60% of fennel extract, organic solvent: 10% -30%, surfactant: 10% -30%, antifreeze: 4% -10%, antimicrobial agent: 0-1% of water and the balance of water, wherein the sum of the weight percentages of the raw materials is 100%.

Optionally, the fennel extract is: adding an extraction solvent into fennel powder with the particle size of 1-5 mm according to a ratio of 1:10(w/v), wherein the extraction solvent is any one or a mixture of more than two of methanol, ethanol, ethyl acetate, chloroform and petroleum ether, and standing at room temperature for 24 hours;

performing ultrasonic extraction at 40 deg.C and frequency of 100Hz for 60min, filtering, extracting the residue for 2 times, mixing the filtrates, and concentrating under reduced pressure to obtain plant extract.

The fennel extract anti-plant virus agent prepared by the invention has the following advantages:

the antiviral agent has obvious activity, can be used for preventing and treating various plant Virus diseases such as Tobacco Mosaic Virus (TMV), Cucumber Mosaic Virus (CMV) and Potato Virus (Potato Virus X, PVX; Potato Virus Y, PVY) on crops such as Tobacco, pepper, tomato, Potato and the like; the fennel extract plant virus resisting agent can improve the activity of defense enzymes in the tobacco body and enhance the disease resistance of plants; the preparation method of the fennel extract plant virus resisting agent is simple and is suitable for popularization and use.

Drawings

The accompanying drawings, which are included to provide a further understanding of the disclosure and are incorporated in and constitute a part of this specification, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure without limiting the disclosure. In the drawings:

FIG. 1 shows the activity change of SOD after spraying fennel extract anti-plant virus agent on tobacco leaves;

FIG. 2 shows the POD activity change after spraying fennel extract on tobacco leaves with anti-plant virus agent.

Detailed Description

The following detailed description of specific embodiments of the present disclosure is provided in connection with the accompanying drawings. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.

Fennel referred to in the present invention is dried ripe fruit of fennel fruit, fennel of family Umbelliferae, Foeniculum vulgare Mill. In autumn, the plant is harvested when the fruit is just ripe, dried in the sun, and the fruit is removed. The fennel extract plant virus resisting agent provided by the invention is prepared by taking a fennel extract as an active substance and adding a certain proportion of auxiliaries into the fennel extract plant virus resisting agent for formulation processing, and the formulation comprises soluble powder, water dispersible granules, soluble liquid and aqueous emulsion

The tests of the inventor prove that the 10% fennel extract soluble powder, the 60% fennel extract soluble powder, the 10% fennel extract water dispersible granules, the 60% fennel extract water dispersible granules, the 10% fennel extract soluble liquid, the 60% fennel extract soluble liquid, the 10% fennel extract water emulsion and the 60% fennel extract water emulsion have good control effects on various plant virus diseases such as Tobacco Mosaic Virus (TMV), Cucumber Mosaic Virus (CMV), potato virus (PVX, PVY) and the like.

The anti-plant virus agent of the invention takes the fennel extract as an effective component, and the extraction method of the fennel extract comprises the following steps:

a: crushing fennel into powder with the particle size of 1-5 mm;

b: adding an extraction solvent at room temperature at a ratio of 1:10(w/v), wherein the extraction solvent is one or a mixture of more than two of methanol, ethanol, ethyl acetate, chloroform and petroleum ether, and standing at room temperature for 24 hours;

c: extracting in an ultrasonic cleaner with frequency of 100Hz at 40 deg.C for 60 min. Filtering, extracting the residue for 2 times, and mixing the filtrates;

d: concentrating the filtrate under reduced pressure to obtain plant extract as effective component of fructus Foeniculi extract for resisting plant virus.

The extract is prepared by extracting perfume plant tissue (such as flower, leaf branch, stem, bark, root, fruit, etc.) without exudate with organic solvent, and the final product contains no original solvent and water; the plant extract mentioned in the invention refers to a substance which is obtained by extracting fennel powder with an organic solvent and concentrating under reduced pressure and does not contain the original organic solvent and water, and the substance is treated by the conventional reduced pressure concentration method in the field.

Unless otherwise specified, the auxiliaries in the present invention are selected from the following classes:

the filler is one or a mixture of more than two of sodium sulfate, ammonium sulfate, attapulgite, kaolin, white carbon, argil and light calcium carbonate;

the dispersant is one or more of alkyl sulfonate, alkyl phenol or fatty alcohol ethylene oxide adduct sulfonate, and phosphate of alkyl phenol and fatty alcohol ethylene oxide adduct;

the stabilizer is one or a mixture of more than two of organic acid, fatty alcohol, diphenyl phosphate, alkyl sulfonate, diisopropyl phosphate and isopropyl phosphate;

the surfactant is one or more of alkyl benzene sulfonate, carboxylate, sulfate, alkyl polyoxyethylene ether, alkylphenol polyoxyethylene ether, polyaryl polyoxyethylene ether and tween;

the organic solvent is one or a mixture of more than two of methanol, ethanol, acetone, ethyl acetate, isopropanol and N, N-dimethylformamide;

the antifreezing agent is one or a mixture of more than two of ethylene glycol, glycerol, urea and glycerol;

the antimicrobial agent is one or a mixture of more than two of 2-hydroxybiphenyl, sorbic acid and benzoic acid.

The preparation method of the soluble powder comprises the following steps: mixing fennel extract, stabilizer, filler, dispersant and surfactant in proportion, dissolving in 0.01mol/L hydrochloric acid solution at room temperature, and stirring to dissolve completely. And drying for 5-10 days at 50-60 ℃ by using a box type dryer to obtain a solid. Pulverizing the solid matter, (the water insoluble filler does not participate in the previous step, and is directly and uniformly mixed with the solid matter), and sieving with a 320-mesh sieve to obtain soluble powder of the fennel extract.

The preparation method of the water dispersible granule comprises the following steps: mixing fructus Foeniculi extract, filler, dispersant and surfactant at a certain ratio, and pulverizing with pulverizer. Adding water, stirring uniformly and granulating. Drying the fennel extract by a dryer at 50-60 ℃ and then sieving the fennel extract to obtain particles with the particle size of 2-4 mm, namely fennel extract water dispersible granules.

The preparation method of the soluble liquid agent comprises the following steps: mixing the fennel extract, the organic solvent, the surfactant, the antifreezing agent and the antimicrobial agent in a mixing kettle in proportion, fully stirring for 30min at the rotating speed of 60-80 r/min, and uniformly stirring to obtain the soluble fennel extract liquid.

The preparation method of the aqueous emulsion comprises the following steps: mixing the fennel extract, the organic solvent, the surfactant and the antimicrobial agent in a mixing kettle in proportion, and stirring until the fennel extract, the organic solvent, the surfactant and the antimicrobial agent are uniformly dissolved; mixing an antifreezing agent and water in proportion, and stirring to dissolve the antifreezing agent and the water into a water phase; slowly adding the water phase into a mixing kettle (the temperature of the mixing kettle is 35-50 ℃), and uniformly stirring for 30min to obtain the fennel extract water emulsion.

In the following examples, "%" means mass percent unless otherwise specified.

For better understanding of the essence of the invention, the technical contents of the invention will be described in detail with examples, but the invention is not limited to these examples.