CN103141524B - The agent of a kind of Chinese Abelia anti cucumber mosaic virus - Google Patents
The agent of a kind of Chinese Abelia anti cucumber mosaic virus Download PDFInfo
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Abstract
The invention discloses a kind of is plant antiviral agent of active ingredient and preparation method thereof with Chinese Abelia (Abelia chinensis R.Br.) extract.This obtained Chinese Abelia anti-plant virus agent adds auxiliary agent with Chinese Abelia extract process for principal component, dosage form is soluble liquid, aqua and microemulsion, can be used for preventing and treating the Cucumber Mosaic Virus (Cucumber mosaic virus, CMV) on the crops such as tobacco, capsicum, tomato, cucumber.Not only anti cucumber mosaic virus disease is effective for anti-plant virus agent of the present invention, and to environment, people, animal, pest natural enemy and other beneficial organism safety.This anti-plant virus agent preparation technology is simple, with low cost, is suitable for applying.
Description
Technical field
The invention belongs to botanical pesticide and application thereof, be specifically related to the agent of a kind of Chinese Abelia anti cucumber mosaic virus.
Background technology
Plant virus a kind ofly infects active cytozoicus pathogen by having of forming of nucleic acid and protein, and according to the 8th report of ICTV (ICTV), the plant virus kind that the current whole world finds reaches 1122.Plant disease caused by plant virus have the title of " plant cancer ", and it is only second to the mycosis original to the extent of injury of crops, and almost often kind of crops are all subject to 2 ~ 3 kinds of viral harm.The loss that the whole world is caused by plant virus every year about 400 hundred million dollars, cucumber mosaic virus (Cucumber mosaic virus, CMV) be host range at most, one of the plant virus of the widest, the most Economic Importance of distribution.
Because plant virus has obligate parasitism to plant cell, the matter and energy place required for virus replication is provided by host plant completely, causes the metabolism of virus and host to combine together; In addition plant lacks complete immune system, and virus, once infect plant, just can be survived, until host is dead indefinite duration in plant cell.Therefore, the medicine of control of plant virosis is only difficult to optionally challenge virus and do not injure host cell, causes the anti-virus formulation research that screening is efficient, single-minded, safe at present to face great difficulty.
At present except taking traditional Anti-virus Disease Breeding, attenuated strain system cross protection, improvement culture technique, insecticide to control to pass except the method such as virus mediator and transgenosis, scientific research personnel also investigated the chemical prevention and control method of some viral diseases of plants.But up to the present, only have sub-fraction anti-plant virus agent to obtain practical application, these preparations are greatly mainly with putting prevention first, and how undesirable drug effect is.Due to people to antivirotic research in medicine or veterinary drug comparatively early, some anti-plant virus agents (as agricultural virazole) are exactly develop from anti-human or zoosis toxic agent.Therefore, further investigated Antiphytoviral natural materials, is conducive to the research of anti-plant virus agent, exploitation and application.
Chinese Abelia (Abelia chinensis R.Br.) belongs to Caprifoliaceae, Abelia plant.Be generally machaka, some place is evergreen shrubs, such as Taiwan Chinese Abelia.Slender lobule multi-branched shrub, up to 2m.Spray by microtriche, bronzing, old branch bark lobe.Leaf to life, sometimes 3 pieces verticillate; Petiole long 1 ?5mm; Blade circle is avette avette to ellipticity, long 2 ?5cm, wide 1 ?3.5cm, the anxious point of tip or short gradually point, basal circular or heart, there is sparse crenature at edge, dredges by undercoat above, below along in the close raw pubescence of base portion of arteries and veins and lateral vein.Cyme is born in sprig top axil, assembles circular cone flowers by most inflorescence; Total bennet is by pubescence, and the fruit phase is smooth; Spend fragrance, tool 3 pairs of bractlets; Calyx tube is cylindrical, by pubescence, slightly flat, tool vertical stripe, and limb 5 splits, and sliver ellipse or the shape of falling ovum Long Circle, be about 5mm, fruit phase curved redness; Corolla white extremely pink, funnel-form.
At present, there is not yet Chinese Abelia is the report that Antiphytoviral toxic agent aspect prepared by raw material.
Summary of the invention
To the object of the invention is to Chinese Abelia (Abelia chinensis R.Br.), for raw material, work out high, nuisanceless, cheap plant resource anti-plant virus agent of Antiphytoviral effect and preparation method thereof.Can be used for preventing and treating the Cucumber Mosaic Virus (Cucumbermosaic virus, CMV) on the crops such as tobacco, capsicum, tomato, cucumber.
In order to realize above-mentioned task, the present invention takes following technical solution:
The agent of a kind of Chinese Abelia anti cucumber mosaic virus, is characterized in that, this obtained Chinese Abelia anti-plant virus agent adds auxiliary agent with Chinese Abelia extract process for principal component, and dosage form is soluble liquid, aqua and microemulsion; Wherein:
Described microemulsion is prepared by following raw materials by weight:
Chinese Abelia extract: 20% ~ 40%, organic solvent: 30% ~ 60%, surfactant: 5% ~ 30%, antifreezing agent: 5% ~ 10%, surplus is water, and the percentage by weight sum of raw material is 100%;
Described soluble liquid is prepared by following raw materials by weight:
Chinese Abelia extract: 20% ~ 40%, organic solvent: 30% ~ 60%, surfactant: 5% ~ 30%, the percentage by weight sum of raw material is 100%;
Described aqua is prepared by following raw materials by weight:
Chinese Abelia extract: 10% ~ 30%, organic solvent: 30% ~ 40%, surfactant: 5% ~ 30%, antifreezing agent: 5% ~ 10%, surplus is water, and the percentage by weight sum of raw material is 100%.
Above-mentioned organic solvent is ethyl acetate, cyclohexanone, methyl oleate, ethanol, methyl alcohol, acetone, the mixture of one or more in dimethyl formamide;
Described emulsifier is that alkyl benzene sulfonate, alkylphenol polyoxyethylene class, multi-styrene phenol APEO, phenethyl phenol polyethenoxy ethers, phenethyl phenol polyethenoxy ether polyoxypropylene ethers and similar substance thereof, this Pan's series, tween, agricultural the moon are from the mixture of one or more in sub-?phosphate;
Described surfactant is one or more mixture in alkyl benzene sulfonate, alkylphenol polyoxyethylene class, phenethyl phenol polyethenoxy ethers, the addition product of phenethyl phenol polyethenoxy ether polyoxypropylene ethers and similar substance, castor oil and oxirane, aralkyl phenol polyethenoxy ether formaldehyde condensation products;
Described antifreezing agent is ethylene glycol or glycerine.
The preparation method of above-mentioned Chinese Abelia anti-plant virus agent, is characterized in that:
The preparation method of described microemulsion is:
A, meal Chinese Abelia acrial part plant tissue cracker being crushed to 1 ~ 5mm mince.
B, to be minced by the meal of steps A gained with extraction, the solvent that lixiviate adopts is ethanol, any one or a few in ethyl acetate, chloroform, benzinum, water; Extracting temperature is 50 ~ 80 DEG C, and extraction time is obtain Chinese Abelia extract, through Vacuum Concentration to being equivalent to 1kg dry powder/kg concentrate in 6 ~ 12 hours.
C, by the concentrate of step B gained and solvent, surfactant, antifreezing agent and water in proportion in mixture still (mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes) stir after namely obtain microemulsion.
The preparation method of described soluble liquid is:
A ~ step B is with the preparation process of microemulsion;
C, by the concentrate of step B gained and solvent, surfactant in proportion in mixture still (mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes), namely obtain soluble liquid after stirring.
The preparation method of described aqua is:
A ~ step B is with the preparation process of microemulsion;
C, by the Chinese Abelia concentrate of step B gained, solvent, surfactant, antifreezing agent and water in proportion in mixture still (mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes) stir after namely obtain aqua.
Chinese Abelia anti cucumber mosaic virus agent made by the present invention has the following advantages:
1. anti-phytoviral activity is remarkable, can be used for the crops such as tobacco, capsicum, tomato, cucumber prevent and treat Cucumber Mosaic Virus (Cucumber mosaic virus, CMV);
2. pair people, animal, pest natural enemy and other beneficial organism safety, environment compatibility is good;
3. biogenic pesticide, uses rear noresidue to poison;
4. aboundresources, preparation method are simple, with low cost, suitable promotes the use of.
Embodiment
In order to better understand essence of an invention, describe the technology contents of invention in detail below in conjunction with embodiment, but invention is not limited to these embodiments.
Chinese Abelia anti-plant virus agent of the present invention is with Chinese Abelia acrial part for raw material, obtains Chinese Abelia extract, add certain proportion auxiliary agent through traditional solvent method lixiviate, through certain process, makes environment-friendly type soluble liquid, aqua or microemulsion.
Described microemulsion is prepared by following raw materials by weight:
Chinese Abelia extract: 20% ~ 40%, organic solvent: 30% ~ 60%, surfactant: 5% ~ 30%, antifreezing agent: 5% ~ 10%, surplus is water, and the percentage by weight sum of raw material is 100%;
Described soluble liquid is prepared by following raw materials by weight:
Chinese Abelia extract: 20% ~ 40%, organic solvent: 30% ~ 60%, surfactant: 5% ~ 30%, the percentage by weight sum of raw material is 100%;
Described aqua is prepared by following raw materials by weight:
Chinese Abelia extract 10% ~ 30%, organic solvent: 30% ~ 40%, surfactant: 5% ~ 30%, antifreezing agent: 5% ~ 10%, surplus is water, and the percentage by weight sum of raw material is 100%;
Described organic solvent is ethyl acetate, cyclohexanone, methyl oleate, ethanol, methyl alcohol, acetone, the mixture of one or more in dimethyl formamide;
Described emulsifier is that alkyl benzene sulfonate, alkylphenol polyoxyethylene class, multi-styrene phenol APEO, phenethyl phenol polyethenoxy ethers, phenethyl phenol polyethenoxy ether polyoxypropylene ethers and similar substance thereof, this Pan's series, tween, agricultural the moon are from the mixture of one or more in sub-?phosphate;
Described surfactant is one or more mixture in alkyl benzene sulfonate, alkylphenol polyoxyethylene class, phenethyl phenol polyethenoxy ethers, the addition product of phenethyl phenol polyethenoxy ether polyoxypropylene ethers and similar substance, castor oil and oxirane, aralkyl phenol polyethenoxy ether formaldehyde condensation products;
Described antifreezing agent is ethylene glycol or glycerine.
The preparation method of above-mentioned soluble liquid, aqua or microemulsion is:
A, meal Chinese Abelia acrial part plant tissue cracker being crushed to 1 ~ 5mm mince.
B, to be minced by the meal of steps A gained and use solvent method lixiviate, the solvent that lixiviate adopts is ethanol, any one or a few in ethyl acetate, chloroform, benzinum, water; Extracting temperature is 50 ~ 80 DEG C, and extraction time is obtain Chinese Abelia extract, through Vacuum Concentration to being equivalent to 1kg dry powder/kg concentrate in 6 ~ 12 hours;
C, by the concentrate of step B gained and solvent, surfactant, antifreezing agent and water in proportion in mixture still, mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes, namely obtains Chinese Abelia Antiphytoviral microemulsion after stirring;
D, by the concentrate of step B gained and solvent, surfactant in proportion in mixture still, mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes, namely obtains Chinese Abelia Antiphytoviral soluble liquid after stirring;
E, by the concentrate of step B gained and solvent, surfactant, antifreezing agent and water in proportion in mixture still, mixture still temperature be 35 ?60 DEG C, mixing time is 30 minutes, namely obtains Chinese Abelia Antiphytoviral aqua after stirring.
Test through inventor proves, the 30% Chinese Abelia microemulsion, 30% Chinese Abelia soluble liquid and the 20% Chinese Abelia aqua that adopt the present invention to prepare have good control efficiency to cucumber mosaic virus (CMV).
It is below the embodiment that inventor provides.
The preparation of embodiment 1:30% Chinese Abelia microemulsion
Take Chinese Abelia concentrate 30kg, be dissolved in 45kg ethyl acetate, then add 15kg calcium dodecyl benzene sulfonate, 5kg styrene acid APEO, ethylene glycol 5kg, mixes under high velocity agitation, is heated to about 40 DEG C; Drip deionized water 30kg under high-speed stirred, control the rate of addition of water, make temperature remain on about 40 DEG C; Water dropwises, and is warming up to 50 DEG C, stirs 1 hour, obtained 20% Chinese Abelia microemulsion 100kg.The stability, outward appearance etc. of preparation meet the requirement of commercial preparation.
The preparation of embodiment 2:30% Chinese Abelia soluble liquid
Take Chinese Abelia concentrate 30kg, be dissolved in 45kg ethyl acetate, add 5kg alkylphenol polyoxyethylene again, 1kg APG A, ethanol polishing is to 100Kg, after mixing, stir 10 ~ 30 minutes with mixing speed 800 ~ 1000 revs/min, Chinese Abelia soluble liquid 100kg can be obtained.The stability, outward appearance etc. of preparation meet the requirement of commercial preparation.
The preparation of embodiment 3:20% Chinese Abelia aqua
Take Chinese Abelia concentrate 20kg, be dissolved in 30kg ethyl acetate, add 5kg alkylphenol polyoxyethylene again, 1kg APG A, 5kg ethylene glycol, water polishing to 100Kg, after mixing, stir 10 ~ 30 minutes with mixing speed 800 ~ 1000 revs/min, Chinese Abelia aqua 100kg can be obtained.The stability, outward appearance etc. of preparation meet the requirement of commercial preparation.
Embodiment 4:3 kind Chinese Abelia antivirotic is to the indoor bioassay of CMV
(1) to the external passivation effect of CMV
Adopt the withered spot method of half leaf to measure, choose that growth is vigorous, growing way is consistent 5 ~ 6 the leaf phase Chenopodium amaranticolor be withered spot host.Take vein as boundary, Zuo Banye inoculates liquid and viral equal-volume mixed liquor, and right half leaf inoculation distilled water and viral equal-volume mixed liquor compare, and virus inoculation concentration is 10 μ g/mL, and passivation time is 5min, rinses inoculation blade surface immediately after inoculation with clear water.Often process inoculation 4 leaves, test repetition 3 times, adds up withered spot number after 3d, calculates inhibiting rate.
Inhibiting rate (%)=(contrast withered spot number-process withered spot number)/contrast withered spot number × 100%
Measurement result is in table 1.
Table 1:3 kind Chinese Abelia antivirotic is to CMV inactivation in vitro
Note: in table, data are the mean value of 3 repetitions; In " preventive effect " hurdle, refer in the level of α=0.05 in significant difference with letter different after ordered series of numbers mark.Following table is same.
(2) to the inhibition of CMV First aggression
Adopt the withered spot method of half leaf, choose growing way consistent, healthy 5 ~ 6 the leaf phase Chenopodium amaranticolor be withered spot host, virus inoculation after smearing liquid 6h, 12h, 24h, blank is clear water process, and 4 leaves are inoculated in each process, repeats 3 times, add up withered spot number after 3d, calculate inhibiting rate.The results are shown in Table 2.
Table 2:3 kind Chinese Abelia antivirotic is to the inhibition of CMV First aggression
(3) to the inhibition of CMV propagation
Adopt the withered spot method of half leaf, choose growing way consistent, healthy 5 ~ 6 the leaf phase Chenopodium amaranticolor be withered spot host, after virus inoculation 6h, 12h, 24h, smear liquid, blank is clear water process, and 4 leaves are inoculated in each process, repeats 3 times, add up withered spot number after 3d, calculate inhibiting rate.The results are shown in Table 3.
The inhibition that table 3:3 kind Chinese Abelia antivirotic is bred CMV
From table 1,2,3,3 kinds of Chinese Abelia antivirotics are to the passivation of CMV, and First aggression, inhibited proliferation are good, apparently higher than contrast medicament moroxydine. second copper.
Embodiment 4:3 kind Chinese Abelia antivirotic is to the pot experiment of CMV
(1) to the protective effect of cucumber
Select 5 ?6 leaf phases, consistent, the healthy and strong cucumber (Cucumis sativus Linn.) of growing way is for examination.Prevention group (dispenser 3 times then inoculate CMV) and control group (only inoculating not dispenser) 2 groups are established in test.With the 200 times of liquid sprayings of 3 kinds of Chinese Abelia antivirotics, contrast medicament moroxydine. second copper 500 times of liquid spraying process cucumber seedlings, every 5d dispenser once, is total to dispenser three times.Inoculate CMV after last dispenser 24h, inoculum density is 1:20 (W/V).After inoculation, 14d, 21d check incidence, statistics disease index and control efficiency.Often process 60 strain cucumber seedlings, test repetition 3 times.Calculate preventive effect.
The incidence of disease=(diseased plant number of getting a haircut everywhere/process total strain number) × 100%
Disease index=∑ (sick progression × diseased plant number)/(the highest sick level × total strain number of each process) × 100
Disease index growth rate=(spray medicine after disease refer to-spray medicine before disease refer to)/spray medicine before disease refer to × 100%
Control efficiency=(contrast disease refers to-process that disease refers to)/contrast disease to refer to × 100%
Protective effect measurement result is as shown in table 4.
Table 4:3 kind Chinese Abelia antivirotic is to the protective effect measurement result of cucumber
(2) to the therapeutic action of cucumber
Select 5 ?6 leaf phases, consistent, the healthy and strong cucumber (Cucumis sativus Linn.) of growing way is for examination.Treatment group (after inoculation CMV dispenser 3 times) and control group (only inoculating not dispenser) 2 groups are established in test.Inoculate malicious source CMV, inoculum density is 1:20 (W/V), and after inoculation, every 5d dispenser once, dispenser three times altogether.Medicament is the 200 times of liquid sprayings of 3 kinds of Chinese Abelia antivirotics, contrast medicament moroxydine. the 500 times of liquid sprayings of second copper.Incidence is checked, statistics disease index and control efficiency during 14d, 21d respectively at last dispenser.Often process 60 strain cucumber seedlings, test repetition 3 times.Calculate preventive effect.Therapeutic action measurement result is as shown in table 5.
Table 5:3 kind Chinese Abelia antivirotic is to the therapeutic action measurement result of cucumber
Results from pot experiment test shows (table 4, table 5), and 3 kinds of Chinese Abelia antivirotics show good prevention and therapy effect to Cucumber Mosaic Virus, and its prevention and therapy effect is all better than contrast medicament, and relative control effect is higher than contrast medicament about 10%.
Embodiment 5:3 kind Chinese Abelia antivirotic is to the drug effect test of field zone of CMV
Community crop is cucumber, and small plot experiment is randomized arrangement, and repeat 3 times, plot area is depending on actual conditions, but area can not be less than 30m
2, experimental field select to require fertility evenly, crop-planting is consistent with managerial skills, state of an illness generation and the extent of injury more even; Protection row will be established between each process and around test region.Blade face conventional spray is carried out, with the moroxydine of 20% with 3 kinds of Chinese Abelia antivirotics, 100,200,400 times of liquid. second copper wetting powder 500 times of liquid carry out blade face conventional spray for contrasting medicament, and establish clear water to contrast; All reagent agents must carry out secondary dilution.The idiopathy initial stage starts to spray medicine, after spray 1 time every 7d, totally 3 times.10d Investigate incidence before first time spray medicine and after last spray medicine and statistics disease index, calculate preventive effect, the results are shown in Table 6.
Table 6:3 kind Chinese Abelia antivirotic control cucumber virus disease drug effect test of field zone
As can be known from the above table, 3 kinds of Chinese Abelia antivirotics have good control efficiency to cucumber mosaic virus, be obviously better than medicament contrast 20% moroxydine when dilution 100,200 times. second copper wetting powder, preventive effect significant difference.
Claims (1)
1. the agent of Chinese Abelia anti cucumber mosaic virus, is characterized in that, this obtained Chinese Abelia anti cucumber mosaic virus agent adds auxiliary agent with Chinese Abelia extract process for principal component, and dosage form is soluble liquid and aqua; Wherein:
Described soluble liquid is prepared by following raw material: take Chinese Abelia concentrate 30kg, be dissolved in 45kg ethyl acetate, add 5kg alkylphenol polyoxyethylene again, 1kg APG A, ethanol polishing is to 100Kg, after mixing, stir 10 ~ 30 minutes with mixing speed 800 ~ 1000 revs/min, can soluble liquid be obtained;
Described aqua is prepared by following raw material: take Chinese Abelia concentrate 20kg, be dissolved in 30kg ethyl acetate, add 5kg alkylphenol polyoxyethylene again, 1kg APG A, 5kg ethylene glycol, water polishing to 100Kg, after mixing, stir 10 ~ 30 minutes with mixing speed 800 ~ 1000 revs/min, can aqua be obtained;
The preparation method of described Chinese Abelia concentrate is:
A, meal Chinese Abelia acrial part plant tissue cracker being crushed to 1 ~ 5mm mince;
B, to be minced by the meal of steps A gained and use solvent method lixiviate, the solvent that lixiviate adopts is ethanol, any one or a few in ethyl acetate, chloroform, benzinum, water; Extracting temperature is 50 ~ 80 DEG C, and extraction time is obtain Chinese Abelia extract, through Vacuum Concentration to being equivalent to 1kg dry powder/kg concentrate in 6 ~ 12 hours.
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| CN101438718A (en) * | 2008-12-30 | 2009-05-27 | 贵州大学 | Medicament for resisting plant virus and formulating method |
| CN102578116A (en) * | 2012-01-20 | 2012-07-18 | 山西德威生化有限责任公司 | Compounded botanical synergistic fungicide containing sophocarpidine and osthol and preparation method of compounded botanical synergistic fungicide |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101438718A (en) * | 2008-12-30 | 2009-05-27 | 贵州大学 | Medicament for resisting plant virus and formulating method |
| CN102578116A (en) * | 2012-01-20 | 2012-07-18 | 山西德威生化有限责任公司 | Compounded botanical synergistic fungicide containing sophocarpidine and osthol and preparation method of compounded botanical synergistic fungicide |
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| 植物源病毒抑制剂VFB活性成分研究及抗TMV植物样品筛选;景炳年;《中国博士学位论文全文数据库农业科技辑》;20121115(第11期);第27页第2.2.1.1节、第28页第2.1.2.1节、第35页表2-1、第43页表2-2、第45页表2-3以及第48页第2.3.2节 * |
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