CN112889833A - Method for preventing and treating plant virus diseases - Google Patents
Method for preventing and treating plant virus diseases Download PDFInfo
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- CN112889833A CN112889833A CN202110133480.9A CN202110133480A CN112889833A CN 112889833 A CN112889833 A CN 112889833A CN 202110133480 A CN202110133480 A CN 202110133480A CN 112889833 A CN112889833 A CN 112889833A
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- compound
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- marine
- natural marine
- sterol
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
- A01N43/54—1,3-Diazines; Hydrogenated 1,3-diazines
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a method for preventing and treating plant virus diseases, in particular to the prevention and treatment by combining a marine sterol extract with an antiviral agent, and the invention also relates to the optimization of a natural marine sterol extraction method. The combined medication result shows that 0.75ug of marine sterol and 1-3mg of antiviral agent can inhibit the tomato spotted wilt virus to the maximum extent, and the effect of fully inhibiting and preventing the virus is achieved.
Description
Technical Field
The invention belongs to an extraction and application method of an antiviral medicament for crops, and particularly relates to an extraction and application method of a preparation for preventing and treating plant virus diseases.
Background
Tomato Spotted Wilt Virus (TSWV) is a harmful organism, named after being found on infected tomatoes firstly, has wide distribution range, can attack 160 dicotyledonous plants and 10 monocotyledonous plants, and crop diseases caused by the infection of the tomato spotted wilt virus are called spotted wilt. Tomato, pepper, tobacco, leaf tobacco, zinnia elegans, lettuce and the like can be systemically infected, and special varieties for resisting TSWV are not bred at present.
The natural sterol medicine is extracted from natural plant sterol, has antiviral activity in different degrees, and is expected to become new generation plant antiviral medicine.
Disclosure of Invention
The invention aims to: extracting natural marine sterol extract, and developing application of natural marine sterol extract in preventing and treating plant virus diseases.
One aspect of the invention relates to a method for controlling plant viral diseases, comprising:
(1) administering an antiviral agent to a diseased tomato;
(2) continuing to apply the natural sterol extract to the tomatoes of step (1);
preferably, the antiviral agent is BTH or ningnanmycin;
preferably, the concentration of the natural sterol extract is 0.75ug/ml, and the application amount is 1 ml/plant;
preferably, the antiviral agent is administered in an amount of 1-3mg per strain;
preferably, the time period for administering the natural sterol extract in step (2) is 1h after administration of the antiviral agent.
Preferably, the extraction method of the natural sterol extract comprises the following steps:
(1) drying 5kg of marine plants to obtain dry powder;
(2) leaching the freeze-dried powder by adopting a normal butane solution, and eluting by using a chloroform-methanol solvent with a certain proportion to obtain a mixture of a compound I and a compound II;
(3) separating the mixture of the first compound and the second compound by a silica gel column, and separating and eluting by ethyl acetate-n-hexane;
(4) adjusting chloroform-methanol ratio for elution to obtain compounds III and IV, and eluting with dichloromethane-acetone;
(5) adjusting the chloroform-methanol ratio again to obtain five and six compounds, and eluting and separating by using dichloromethane-acetone. The first to sixth compounds are natural marine sterol.
Preferably, the chloroform-methanol ratios in steps (2), (4) and (5) are 10:1,20:1 and 15:1, respectively;
preferably, the ethyl acetate-n-hexane ratio in the step (3) is 1: 3;
preferably, the ratio of dichloromethane-acetone in step (4) (5) is 1:2,1: 3;
preferably, the marine organism is herba Zosterae Marinae, Corallium japonicum Kishinouye, and herba Lysimachiae Fortunei.
Preferably, compounds one to five have the following structures, wherein compound one R1 ═ H, R2 ═ OH, compound two R1 ═ OH, R2 ═ H, compound three R1 ═ R2 ═ H, compound four R1 ═ OAc, R2 ═ H, compound five R1 ═ OH, and compound R2 ═ H.
The structure of compound six is shown below:
advantageous effects
The invention adopts the combination of the natural marine sterol extract and the chemical agent to prevent and treat the tomato virus diseases, has better prevention and treatment effect compared with the single agent, and can reduce the side effect and pollution of the chemical agent to crops to a certain extent.
The invention further optimizes the parameters of the extraction method of the marine sterol, and improves the extraction and separation effects.
Detailed Description
Example 1:
(1) taking 5kg of coral, and drying to obtain dry powder;
(2) leaching the freeze-dried powder by using a normal butane solution, and eluting by using a chloroform-methanol solvent (10:1) to obtain a mixture of a compound I and a compound II;
(3) separating the mixture of the first compound and the second compound by a silica gel column, and separating and eluting by ethyl acetate-n-hexane (1: 3);
(4) adjusting chloroform-methanol ratio (20:1) to obtain compounds III and IV, and eluting with dichloromethane-acetone (1: 2);
(5) adjusting the chloroform-methanol ratio again (15:1) to obtain five and six compounds, and eluting and separating by using dichloromethane-acetone (1: 3).
The first to sixth compounds and ningnanmycin are respectively applied in a combined mode to prevent and treat the tomato spotted wilt virus, the application concentration/application amount of the compounds is 1ug/ml, 1 ml/plant, the application amount of the ningnanmycin is 1 mg/plant, 2ml of the compounds and 2mg of the ningnanmycin are respectively applied to a control group, tomato stems and leaves are observed for 6 hours, 12 hours, 24 hours, 48 hours and 72 hours after application (wherein the results show that no obvious effect is observed, + shows that a slight effect is observed, + shows that an effect is obvious, and + + + shows that a basic cure effect is achieved), and the results are shown in the following table:
TABLE 1 controlling effect of natural marine sterols in combination with ningnanmycin
From the experimental results, the natural marine sterol can play a role in preventing and treating tomato spotted wilt virus, the effect time of the natural marine sterol is equivalent to that of ningnanmycin, and the safety problem of chemical drugs is avoided. Compared with the single chemical medicament, the combination of the natural sterol and the antiviral medicament has better effect on preventing the tomato spotted wilt virus, which is probably related to different antiviral mechanisms of the two antiviral medicaments.
On the basis of the above experiment, further exploring the optimum concentration of the combined application, selecting the compound two which performs the best in the early stage, applying 1ml of the compound two with the concentration of 0.25ug/ml, 0.5ug/ml, 0.75ug/ml and 1ug/ml respectively, observing the growth condition by taking the infected tomato which is not applied as a negative control, and showing the following results (-indicating no significant effect observed, + indicating slight effect, + indicating significant effect, + indicating basic cure):
TABLE 2 comparison of control effects for different application rates
As can be seen from the above, the application amount of 1ml, 0.75ug/ml can substantially meet the demand, and further, the application amount is increased without significant improvement, preferably 0.75ug/ml in view of the application cost.
On the basis of the above, the administration amount of the antiviral drugs of the combination is further considered, and the growth condition is observed by using 0.75ug/ml of compound two 1ml and combining 0mg, 1mg, 2mg, 3mg and 5mg of Zhouqi for infected tomatoes, and the results are as follows (-indicating no obvious effect is observed, + indicating a slight effect, + indicating an obvious effect, and + + + indicating a basic cure):
| treatment/time | 12h | 24h | 48h | 96h | 120h |
| 0mg ningnanmycin | - | - | + | + | ++ |
| 1mg ningnanmycin | - | + | + | ++ | ++ |
| 2mg ningnanmycin | - | + | + | ++ | ++ |
| 3mg ningnanmycin | + | + | ++ | ++ | +++ |
| 5mg ningnanmycin | + | + | ++ | ++ | +++ |
From the above, the compound II of 0.75ug/ml and the ningnanmycin of 3mg can realize the economic and maximum tomato spotted wilt virus prevention and treatment effect.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A method of controlling viral diseases in plants, comprising: natural marine sterols are administered.
2. A method according to claim 1 for controlling viral diseases in plants, characterized in that: antiviral agents are administered prior to administration of the natural marine sterols.
3. The method for controlling a plant viral disease according to claim 2, wherein: the plant viral disease is caused by tomato spotted wilt virus.
4. A method according to claim 2 for controlling viral diseases in plants, characterized in that: the natural marine sterol is selected from compound one to compound six, compound one to compound five are shown in the left formula, compound six is shown in the right formula, wherein compound one R1 ═ H, R2 ═ OH, compound two R1 ═ OH, R2 ═ H, compound three R1 ═ R2 ═ H, compound four R1 ═ OAc, R2 ═ H, compound five R1 ═ OH, R2 ═ H
5. A method according to claim 2 for controlling viral diseases in plants, characterized in that: the antiviral agent is BTH or ningnanmycin.
6. A method according to claim 2 for controlling viral diseases in plants, characterized in that: the dosage of antiviral agent is 1-3mg, the concentration of natural marine sterol is 0.75ug/ml, the dosage is 1ml, and the application interval is 1 h.
7. A method according to claim 2 for controlling viral diseases in plants, characterized in that: the extraction method of the natural marine sterol comprises the following steps:
(1) drying 5kg of marine plants to obtain dry powder;
(2) leaching the freeze-dried powder by adopting a normal butane solution, and eluting by using a chloroform-methanol solvent with a certain proportion to obtain a mixture of a compound I and a compound II;
(3) separating the mixture of the first compound and the second compound by a silica gel column, and separating and eluting by ethyl acetate-n-hexane;
(4) adjusting chloroform-methanol ratio for elution to obtain compounds III and IV, and eluting with dichloromethane-acetone;
(5) adjusting the chloroform-methanol ratio again to obtain five and six compounds, and eluting and separating by using dichloromethane-acetone.
8. The method of extracting natural marine sterols according to claim 7, wherein: in the steps (2), (4) and (5), the ratio of chloroform to methanol is 10:1,20:1 and 15:1 respectively, and the ratio of ethyl acetate to n-hexane in the step (3) is 1: 3.
9. The method of extracting natural marine sterols according to claim 7, wherein: the ratio of dichloromethane to acetone in the steps (4) and (5) is 1:2 and 1: 3.
10. The method of extracting natural marine sterols according to claim 7, wherein: the marine organism is herba Zosterae Marinae, Corallium japonicum Kishinouye, and herba Lysimachiae Fortunei.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110133480.9A CN112889833A (en) | 2021-02-01 | 2021-02-01 | Method for preventing and treating plant virus diseases |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110133480.9A CN112889833A (en) | 2021-02-01 | 2021-02-01 | Method for preventing and treating plant virus diseases |
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| CN112889833A true CN112889833A (en) | 2021-06-04 |
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Citations (8)
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|---|---|---|---|---|
| CN101560236A (en) * | 2009-05-22 | 2009-10-21 | 中山大学 | Method for synthesizing marine polyhydroxyl sterol (25R)-5alpha -cholest-3 beta, 5alpha, 6beta, 26-tetrol |
| CN102742475A (en) * | 2011-05-04 | 2012-10-24 | 蔡英明 | Plant disease classification prevention method |
| CN102812969A (en) * | 2012-08-29 | 2012-12-12 | 陕西上格之路生物科学有限公司 | Botanical antiviral agent, and preparation method and application thereof |
| CN102838648A (en) * | 2012-08-29 | 2012-12-26 | 陕西上格之路生物科学有限公司 | Steroid enol compound as well as preparation method thereof and application thereof |
| CN104004042A (en) * | 2014-05-17 | 2014-08-27 | 中国海洋大学 | Steroidal compound as well as preparation method and application of steroidal compound taken as antiviral agent |
| CN104262443A (en) * | 2014-09-02 | 2015-01-07 | 广西壮族自治区海洋研究所 | Preparation method of polyhydroxy sterol monomeric compound and application |
| CN107568234A (en) * | 2017-09-30 | 2018-01-12 | 京博农化科技股份有限公司 | A kind of antiviral composition |
| CN111418595A (en) * | 2020-04-27 | 2020-07-17 | 中国科学院昆明植物研究所 | Application of C21 steroid compounds 1-8 in paniculate swallowwort root in preparing anti-TSWV active product |
-
2021
- 2021-02-01 CN CN202110133480.9A patent/CN112889833A/en active Pending
Patent Citations (8)
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|---|---|---|---|---|
| CN101560236A (en) * | 2009-05-22 | 2009-10-21 | 中山大学 | Method for synthesizing marine polyhydroxyl sterol (25R)-5alpha -cholest-3 beta, 5alpha, 6beta, 26-tetrol |
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| CN107568234A (en) * | 2017-09-30 | 2018-01-12 | 京博农化科技股份有限公司 | A kind of antiviral composition |
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Application publication date: 20210604 |