CN104738102B - Chinese gooseberry root bark extract is used for the application for preparing anti-plant virus agent - Google Patents
Chinese gooseberry root bark extract is used for the application for preparing anti-plant virus agent Download PDFInfo
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Abstract
It is used for the application for preparing anti-plant virus agent the invention discloses Chinese gooseberry root bark extract, take Chinese gooseberry root skin, crushed after drying, through certain process, it is prepared into the antiviral soluble liquid of Kiwi berry root skin, aqua and microemulsion, available for preventing and treating tobacco mosaic virus disease (Tobacco mosaic virus in tobacco, capsicum, tomato, cucurbita pepo, potatoes and other crops, TMV), Cucumber Mosaic Virus (Cucumber mosaic virus,) and potato virus disease (Potato Virus X, PVX CMV；Potato Virus Y, PVY) etc. various plants virosis；Not only antiviral effect is good for the anti-plant virus agent of the present invention, and to environment, people, animal, pest natural enemy and other beneficial organisms safety.Anti-plant virus agent preparation technology is simple, and cost is cheap, suitable popularization and application.
Description
Technical field
The invention belongs to plant and its application field, and in particular to Chinese gooseberry root bark extract is used for Antiphytoviral
Application.
Background technology
Plant virus be it is a kind of by nucleic acid and protein form have infect activity cytozoicus pathogen, according to state
8th report of the border virus taxis committee (ICTV), the plant virus species that the whole world is found at present is up to 1122.Plant
Plant disease caused by virus is known as the title of " plant cancer ", and its extent of injury to crops is only second to the nosomycosis original,
Almost every kind of crops are all by 2~3 kinds of viral harm.It is beautiful that the whole world loses about 400 hundred million as caused by plant virus every year
Harm is just more than 100,000,000 dollars caused by member, only tobacco mosaic virus (TMV) (Tobacco mosaic virus, TMV).
Because plant virus has strict parasitics, it, which is colonized in, carries out nucleic acid in plant cell even nucleus
The duplication of (RNA or DNA) and the synthesis of protein coat, form new virion.This just determines the anti-of the viroses of plant
It is big to control difficulty, general medicament does not have effect.Early 20th century, Allard reported the inhibiting substances that TMV in Phytolacca acinosa juice be present, afterwards
The emphasis of preventing and treating plant virus is increasingly turned to natural plant extract direction.Particularly since the nineties in last century, Ren Menlu
The continuous inhibiting substances that many Antiphytovirals are have found from natural products, it is natural for efficient, single-minded, the safe plant virus of exploitation
Inhibitor provides the foundation.
Chinese gooseberry Actinidia chinensis Planch. are Actinidiaceae actinidia, and its root is also known as
Chinese actinidia root, it is Chinese unique wheat, is mainly distributed on Chinese Shaanxi, Jiangxi, Hunan, Hubei, Fujian, Zhejiang, Henan, Anhui etc.
Save.Chinese gooseberry complete stool is pharmaceutically acceptable, and the traditional Chinese medical science thinks its root and root skin is cold in nature, bitter and puckery flavor, has activating blood circulation and reducing swelling, wind-dispelling profit
The effect of wet, reducing fever and causing diuresis, radiating hemostasis, available for treatment hepatitis, rheumatic arthritis, dysentery, filariasis, stomach cancer, mammary gland
The illnesss such as cancer.Modern age pharmaceutical research finds that Chinese gooseberry root has scavenging capacity oxygen radical, antiviral, liver protection, suppression
Growth of tumour cell, improve the functions such as immunity.
It has been reported that Chinese gooseberry root chemical composition be mainly flavones, triterpene, steroidal, anthraquinone, phenolic acid, carbohydrate and its
His polytype compound, wherein based on triterpenes components.The pharmacological activity for the reactive compound having found mainly shows
In terms of medicinal activity, such as antitumor, enhancing body's immunity, protection liver, anti-oxidant, removing free radical, anti-colyliform disease
The effect of malicious.Up to the present, on Chinese gooseberry root bark extract and its metabolite to disease, worm, grass in agricultural production
Harmful effect has no report.
The content of the invention
It is an object of the invention to Chinese gooseberry (Actinidia chinensis Planch.) root bark extract
For raw material, high, nuisanceless, the cheap plant source anti-plant virus agent of Antiphytoviral effect is worked out.
To achieve the above object, the technical scheme taken of the present invention is：
Chinese gooseberry root bark extract is used for the application for preparing anti-plant virus agent.
Specifically, described Chinese gooseberry root bark extract is to be obtained after Chinese gooseberry root skin crushed material is carried out into alcohol extracting
The material arrived.
More specifically, the particle diameter of described Chinese gooseberry root skin crushed material is 1~5mm, and the solvent that alcohol extracting uses is second
Alcohol, alcohol extracting temperature are 50~70 DEG C.
Further, the solvent that alcohol extracting uses is the ethanol of mass fraction 95%, and the alcohol extracting time is 18~20 hours.
Meanwhile described plant virus includes tobacco mosaic virus (TMV), cucumber mosaic virus and Potyvirus.
A kind of anti-plant virus agent, the formulation of the anti-plant virus agent is microemulsion, and described microemulsion is by following raw material
Prepare by weight percentage：
Chinese gooseberry root bark extract：15%~30%, organic solvent：0~55%, surfactant：5%~
20%, antifreezing agent：5%~10%, surplus is water, and the percentage by weight sum of raw material is 100%.
Or the formulation of the anti-plant virus agent is soluble liquid, described soluble liquid is by following raw material by weight
Percentage is measured to prepare：
Chinese gooseberry root bark extract 15%~30%, organic solvent：0%~55%, surfactant：5%~
30%, surplus is ethanol, and the percentage by weight sum of raw material is 100%.
Or the formulation of the anti-plant virus agent is aqua, described aqua is prepared by following raw materials by weight：
Chinese gooseberry root bark extract：10%~20%, organic solvent：0%~40%, surfactant：5%~
30%, antifreezing agent：5%~10%, surplus is water, and the percentage by weight sum of raw material is 100%.
Kiwi berry root skin anti-plant virus agent made by the present invention has advantages below：
(1) present invention has found that the root skin of Chinese gooseberry is extracted by carrying out research to the Kiwi extract of different cultivars
Thing is notable to the preventive effect of plant virus, hence it is evident that the preventive effect better than other kind Kiwi extracts to plant virus；
(2) in addition, inventor has also carried out Antiphytoviral work to the extract of the different parts of Chinese kiwi fruit plant
Property research, find only have Chinese gooseberry root skin position extract it is notable to the preventive effect of plant virus, other positions are very
Extremely do not have anti-phytoviral activity；
(3) the anti-phytoviral activity wide spectrum of Chinese gooseberry extract of the invention, available for tobacco, capsicum, tomato,
Tobacco mosaic virus disease (Tobacco mosaic virus, TMV), cucumber mosaic virus are prevented and treated on cucurbita pepo, potatoes and other crops
Viral disease (Cucumber mosaic virus, CMV) and potato virus disease (Potato Virus X, PVX；Potato Virus
Y, PVY) etc. various plants virosis.
Embodiment
The plant source anti-plant virus agent of the present invention, using perennial Chinese gooseberry root bark extract as active material, adds
Enter certain proportion auxiliary agent and carry out formulation, environment-friendly type preparation microemulsion, aqua or soluble liquid, institute of the present invention can be made
The Chinese gooseberry root bark extract stated is the material after alcohol extracting, and optional alcohol extraction solvent is ethanol, and the plant extracts is resisted
The effect of virus has no adverse effects.
Plant virus of the present invention includes：Tobacco mosaic virus disease (Tobacco mosaic virus, TMV), Huang
Melon mosaic virus (Cucumber mosaic virus, CMV) and potato virus disease (Potato Virus X, PVX；
Potato Virus Y, PVY).
The extracting method of Chinese gooseberry root bark extract of the present invention includes：
A. the coarse powder for Chinese gooseberry root skin being crushed to 1~5mm with plant tissue pulverizer minces；
B. it is 50~70 DEG C the coarse powder obtained by step A to be minced with 95% alcohol steep, Extracting temperature, extraction time 18
~20 hours, extract was through being concentrated in vacuo to equivalent to 1kg dry powder/kg concentrates.
Organic solvent of the present invention is ethyl acetate, cyclohexanone, methyl oleate, ethanol, methanol, acetone and dimethyl
Mixture more than one or both of formamide；
Described emulsifying agent is alkyl benzene sulfonate, APES class, multi-styrene phenol polyoxyethylene
Ether, phenethyl phenol polyethenoxy ether class, phenethyl phenol polyethenoxy ether polyoxypropylene ethers and the like matter, this Pan series,
Mixture more than one or both of tween and agricultural anion-phosphate；
Described surfactant is alkyl benzene sulfonate, APES class, phenethyl phenol polyoxyethylene
The addition product and virtue of ethers, phenethyl phenol polyethenoxy ether polyoxypropylene ethers and the like matter, castor oil and oxirane
Mixture more than one or both of APES formaldehyde condensation product；
Described antifreezing agent is ethylene glycol or glycerine.
In order to clear and definite Chinese gooseberry root skin antiviral activity particularity and further explore its active active component,
Inventor has made the work of several lower three aspects in advance.
First, Chinese gooseberry root skin belongs to the difference of other plant antiviral activity together：
Inventor compares the root skin of Chinese gooseberry and other several actinidias nearer with its affiliation
The antiviral activity of extract, as a result shows, the antiviral activity of Chinese gooseberry root bark extract is significantly better than other kinds.
It the results are shown in Table 1.
Several Chinese actinidia root bark extracts of table 1 are to TMV inactivation in vitro
Note：1st, reagent agent is 20% root bark extract microemulsion；
2nd, data are the average value of 3 repetitions in table；In " preventive effect " column, with different letters after ordered series of numbers mark refer to α=
It is in significant difference in 0.05 level.
2nd, the determination at Chinese gooseberry antiviral activity position：
Inventor determines external passivation activity of the Chinese gooseberry different parts extract to TMV, the results showed that, it is different
The antiviral activity of extractive part is widely different, and under same concentrations, root bark extract has significant activity, root, stem, Ye Jun
Activity is not shown, and complete stool shows certain activity, may be also due to the effect (table 2) of antiviral activity in root skin.Accordingly,
Inventor speculates that antiviral activity concentrates on root skin position in Chinese gooseberry, therefore, present invention determine that Chinese gooseberry root skin
Extract is the primary raw material of antiviral activity.
The Chinese gooseberry different parts extract of table 2 is to TMV inactivation in vitro
Note：1st, "-" represents not show passivation activity.
2nd, data are the average value of 3 repetitions in table；In " preventive effect " column, with different letters after ordered series of numbers mark refer to α=
It is in significant difference in 0.05 level.
3rd, the broad spectrum activity of Chinese gooseberry root bark extract antiviral activity：
Inventor determines external passivation activity of the Chinese gooseberry root bark extract to several frequently seen plants virosis, knot
Fruit shows that it shows certain activity to TMV, CMV, PVX and PVY, this explanation, contains wide spectrum in Chinese gooseberry root skin
The anti-phytoviral activity composition of property.
Inactivation in vitro of the Chinese gooseberry root bark extract of table 3 to several plant virosis
Note：Data are the average value of 3 repetitions in table；In " preventive effect " column, with different letters after ordered series of numbers mark refer to α=
It is in significant difference in 0.05 level, it is as follows.
On the basis of being tested more than, in order to further develop the antiviral activity system of Chinese gooseberry root bark extract
Agent, microemulsion, soluble liquid and aqua is respectively prepared.Experiment through inventor proves, 20% Kiwi berry root skin micro emulsion
Agent, 20% Chinese actinidia root skin soluble liquid and 10% Chinese actinidia root severe edema due to hypofunction of the spleen agent have good to tobacco mosaic virus (TMV) (TMV)
Prevention effect, wherein optimal with the prevention effect of 20% Kiwi berry root skin microemulsion.In order to be better understood from the essence of invention, under
Face describes the technology contents of invention in detail with embodiment, but invention is not limited to these embodiments.
4th, specific preparation preparating example：
Embodiment 1：The preparation of 20% Kiwi berry root skin microemulsion
Kiwi berry root skin concentrate 20kg is weighed, is dissolved in 30kg ethyl acetate, adds 15kg detergent alkylate sulphurs
Sour calcium, 5kg styrene acid APEOs, 5kg ethylene glycol, mixes, is heated to 40 DEG C or so under high velocity agitation；High-speed stirred
Lower dropwise addition deionized water 25kg, controls the rate of addition of water, keeps the temperature at 40 DEG C or so；Water is added dropwise, and is warming up to 50
DEG C, stir 1 hour, 20% Kiwi berry root skin microemulsion 100kg is made.The stability of preparation, outward appearance etc. meet commercial preparation
It is required that.
Embodiment 2：The preparation of 20% Chinese actinidia root skin soluble liquid
Kiwi berry root skin concentrate 20kg is weighed, is dissolved in 30kg ethyl acetate, adds 5kg alkylphenol-polyethenoxies
Ether, 1kg APG A, ethanol polishing to 100kg, after being well mixed, with 800~1000 revs/min of stirrings 10 of mixing speed
~30 minutes, you can Chinese actinidia root skin soluble liquid 100kg is made.The stability of preparation, outward appearance etc. meet commercial preparation
It is required that.
Embodiment 3：The preparation of 10% Chinese actinidia root severe edema due to hypofunction of the spleen agent
Kiwi berry root skin concentrate 10kg is weighed, is dissolved in 30kg ethyl acetate, adds 5kg alkylphenol-polyethenoxies
Ether, 1kg APGs A, 5kg ethylene glycol, water polishing to 100kg, after being well mixed, with 800~1000 revs/min of mixing speed
Clock stirs 10~30 minutes, you can Chinese actinidia root skin soluble liquid 100kg is made.The stability of preparation, outward appearance etc. meet business
The requirement of product preparation.
5th, indoor bioassay of 3 kinds of Kiwi berry root skin antivirotics to TMV：
(1) to the external passivation effects of TMV
Determined using half leaf withered spot method, choose 5~6 leaf phase Nicotiana glutinosa (Nicotiana that growth is vigorous, growing way is consistent
Glutinosa) it is withered spot host.Using vein as boundary, left half leaf inoculation decoction and the isometric mixed liquor of virus, right half leaf inoculation are steamed
Distilled water compares with viral isometric mixed liquor, and virus inoculation concentration is 10 μ g/mL, passivation time 5min, after inoculation immediately
Inoculation blade surface is rinsed with clear water.4 leaves of often processing inoculation, experiment are repeated 3 times, and withered spot number is counted after 3d, calculates and suppresses
Rate.
Inhibiting rate (%)=(control withered spot number-processing withered spot number)/control withered spot number × 100
Measurement result is shown in Table 4.
43 kinds of Kiwi berry root skin antivirotics of table are to TMV inactivation in vitro
(2) to the inhibition of TMV First aggressions
Using half leaf withered spot method, it is withered spot host to choose 5~6 leaf phase Nicotiana glutinosas that growing way is consistent, healthy, is smearing decoction
Virus inoculation after 6h, 12h, 24h, blank control are clear water processing, 4 leaves of each processing inoculation, are repeated 3 times, are counted after 3d
Withered spot number, calculate inhibiting rate.It the results are shown in Table 5.
Inhibition of the 53 kinds of Kiwi berry root skin antivirotics of table to TMV First aggressions
(3) to the inhibition of TMV propagation
Using half leaf withered spot method, it is withered spot host to choose 5~6 leaf phase Nicotiana glutinosas that growing way is consistent, healthy, in virus inoculation
Decoction is smeared after 6h, 12h, 24h, blank control is clear water processing, 4 leaves of each processing inoculation, is repeated 3 times, is counted after 3d
Withered spot number, calculate inhibiting rate.It the results are shown in Table 6.
The inhibition that 63 kinds of Kiwi berry root skin antivirotics of table are bred to TMV
From table 4,5,6, passivation of 3 kinds of Kiwi berry root skin antivirotics to TMV, First aggression, proliferation inhibiting effect are good
It is good, wherein, the effect of 20% Kiwi berry root skin microemulsion is better than other two kinds of formulations, hence it is evident that higher than comparison medicament moroxydine second
Copper.
(4) to TMV pot experiment
1) to the protective effect of common cigarette
The 5-6 leaf phases are selected, the common cigarette (Nicotiana tabacum) that growing way is consistent, healthy and strong is for examination.Experiment sets prevention group
2 groups of (dispenser 3 times then be inoculated with TMV) and control group (only inoculation not dispenser).Sprayed with 3 kinds of Kiwi berry root skin antivirotics, 200 times of liquid
Mist, comparison medicament moroxydine second 500 times of common cigarettes of liquid spraying treatment of copper, per 5d dispensers once, common dispenser is three times.For the last time
TMV, inoculum density 1 are inoculated with after dispenser 24h：20(W/V).After inoculation 14d, 21d check incidence, count disease index and
Prevention effect.60 plants of tobacco seedlings are often handled, experiment is repeated 3 times.Calculate preventive effect.
The incidence of disease=(diseased plant number/processing total strain number of getting a haircut everywhere) × 100%
Disease index=∑ (sick series × diseased plant number)/(highest disease level × respectively handle total strain number) × 100
Disease refers to disease index growth rate=(disease refers to after spray-before spraying) disease refers to × 100% before/spray
Prevention effect=(control disease refer to-handles disease and referred to)/control disease refers to × 100%
Protective effect measurement result is as shown in table 7.
Protective effect measurement result of the 73 kinds of Kiwi berry root skin antivirotics of table to common cigarette
2) to the therapeutic action of common cigarette
The 5-6 leaf phases are selected, the common cigarette (Nicotiana tabacum) that growing way is consistent, healthy and strong is for examination.Experiment sets treatment group
(dispenser 3 times after inoculation TMV) and 2 groups of control group (only inoculation not dispenser).It is inoculated with malicious source TMV, inoculum density 1：20 (W/V),
Once, common dispenser is three times for every 5d dispensers after inoculation.Medicament is that 3 kinds of Kiwi berry root skin antivirotics, 200 times of liquid are sprayed, comparison medicament
The 500 times of liquid sprayings of moroxydine second copper.Incidence is checked when 14d, 21d of last time dispenser, counts disease index
And prevention effect.60 plants of tobacco seedlings are often handled, experiment is repeated 3 times.Calculate preventive effect.Therapeutic action measurement result is as shown in table 8.
Therapeutic action measurement result of the 83 kinds of Kiwi berry root skin antivirotics of table to common cigarette
Results from pot experiment test shows (table 7, table 8) that 3 kinds of Kiwi berry root skin antivirotics are shown to tobacco mosaic virus disease
Good prevention and treatment effect, it is protected and therapeutic effect is superior to comparison medicament, wherein, 20% Kiwi berry root skin microemulsion
Protective effect relative control effect is higher than comparison medicament 25% or so.
6th, drug effect test of field zone of 3 kinds of Kiwi berry root skin antivirotics to TMV：
Plot experiment is random alignment, is repeated 3 times, and plot area is depending on actual conditions, but area can not be less than 30
㎡, experimental field selection require that fertility is uniform, crop-planting is consistent with managerial skills, and the state of an illness occurs and the extent of injury is than more uniform；
Protection row is set between each processing and around trial zone.Blade face is carried out with 3 kinds of Kiwi berry root skin antivirotics, 100,200,400 times of liquid
Conventional spray, blade face conventional spray is carried out by comparison medicament of 20% moroxydine 500 times of liquid of second copper wettable powder, and set
Clear water compares；All reagent agents must carry out secondary dilution.Idiopathy starts to spray initial stage, after sprayed 1 time, totally 3 times every 7d.
10d Investigate incidences and statistics disease index before spraying for the first time and after last time spray, calculate preventive effect, the results are shown in Table 9.
93 kinds of Kiwi berry root skin antivirotic preventing and treating tobacco virus drug effect test of field zone of table
As can be known from the above table, 3 kinds of Kiwi berry root skin antivirotics have good prevention effect to tobacco mosaic virus (TMV), dilute
Medicament comparison virus A, preventive effect significant difference are substantially better than in the case of releasing 100,200,400 times, and 20% Kiwi berry root skin is micro-
The average preventive effect of emulsion is better than 20% Chinese actinidia root skin soluble liquid and the average preventive effect of 10% Chinese actinidia root severe edema due to hypofunction of the spleen agent, it is seen that
Chinese actinidia root bark extract is prepared into microemulsion advantageously in the preventing and treating to plant virus.
Claims (4)
1. Chinese gooseberry root bark extract is used for the application for preparing anti-plant virus agent；
Described Chinese gooseberry root bark extract is the material that will be obtained after the progress alcohol extracting of Chinese gooseberry root skin crushed material；
The particle diameter of described Chinese gooseberry root skin crushed material is 1~5mm, and the solvent that alcohol extracting uses is ethanol, and alcohol extracting temperature is
50~70 DEG C；The solvent that alcohol extracting uses is the ethanol of mass fraction 95%, and the alcohol extracting time is 18~20 hours；
Described plant virus includes tobacco mosaic virus (TMV), cucumber mosaic virus and Potyvirus.
A kind of 2. anti-plant virus agent, it is characterised in that the formulation of the anti-plant virus agent is microemulsion, described microemulsion by
Following raw materials by weight is prepared：
Chinese gooseberry root bark extract：15%~30%, organic solvent：0~55%, surfactant：5%~20%, prevent
Freeze agent：5%~10%, surplus is water, and the percentage by weight sum of raw material is 100%.
3. a kind of anti-plant virus agent, it is characterised in that the formulation of the anti-plant virus agent is soluble liquid, and described is solvable
Property liquor is prepared by following raw materials by weight：
Chinese gooseberry root bark extract 15%~30%, organic solvent：0%~55%, surfactant：5%~30%, it is remaining
Measure as ethanol, the percentage by weight sum of raw material is 100%.
4. a kind of anti-plant virus agent, it is characterised in that the formulation of the anti-plant virus agent is aqua, and described aqua is by following
Raw materials by weight is prepared：
Chinese gooseberry root bark extract：10%~20%, organic solvent：0%~40%, surfactant：5%~30%,
Antifreezing agent：5%~10%, surplus is water, and the percentage by weight sum of raw material is 100%.
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| CN105145675A (en) * | 2015-09-02 | 2015-12-16 | 孙凯 | Poultry raising house air disinfectant and preparation method thereof |
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| CN111657305B (en) * | 2020-05-06 | 2021-09-07 | 西北农林科技大学 | Application of fennel extract in preparation of anti-plant virus agent and anti-plant virus agent |
| CN112369437B (en) * | 2020-10-23 | 2021-11-23 | 江西省林业科学院 | Method and medicament for treating citrus huanglongbing |
| CN115251053B (en) * | 2022-07-19 | 2023-09-26 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Sclareol plant antiviral preparation and application thereof |
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