CN114982790A - Microemulsion containing tobacco cis-abienol and application thereof - Google Patents
Microemulsion containing tobacco cis-abienol and application thereof Download PDFInfo
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- CN114982790A CN114982790A CN202210845060.8A CN202210845060A CN114982790A CN 114982790 A CN114982790 A CN 114982790A CN 202210845060 A CN202210845060 A CN 202210845060A CN 114982790 A CN114982790 A CN 114982790A
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
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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
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/06—Oxygen or sulfur directly attached to a cycloaliphatic ring system
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/38—Solanaceae [Potato family], e.g. nightshade, tomato, tobacco or chilli pepper
- A01N65/385—Tobacco
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses a microemulsion containing tobacco cis-abienol, which comprises the following components in percentage by weight: 2-4% of cis-fir alcohol extract, 30-40% of ethanol, 10-20% of fatty alcohol-polyoxyethylene ether, 3-7% of sodium dodecyl sulfate and the balance of water. The invention takes the alcohol extract of the cis-fir of the tobacco as the raw material, only takes ethanol as the solvent, and selects the non-toxic or low-toxic emulsifier and the coemulsifier as the auxiliary agent to prepare the microemulsion, the process is simple, the industrialized production is convenient, and the environment is not polluted; the tobacco cis-abienol microemulsion is used for preventing and treating potato virus Y, tobacco mosaic disease and cucumber mosaic disease, has better prevention effect than amino-oligosaccharin, and enriches the types of Chinese plant-derived virus preparations.
Description
Technical Field
The invention relates to a microemulsion containing tobacco cis-abienol and application thereof, belonging to the technical field of application of plant secondary metabolites.
Background
Cis-abienol is a main labdane compound secreted by tobacco glandular hairs, is named as being originally found in fir oleoresin canada (GRAY et al, 1964), is mainly found in aromatic tobacco and cigar, is an important precursor of tobacco aroma components, and has various biological activities of bacteriostasis, insect resistance, virus resistance and the like. SEO et al (2012)863-864 applying cis-abienol to roots of tobacco, tomato and Arabidopsis infected with blight, the results indicate that cis-abienol can slow down the occurrence of blight by inhibiting the growth of bacteria; cis-abienol can be used as an endogenous signal to rapidly increase the waf1 protein in tobacco leaves infected with Tobacco Mosaic Virus (TMV), so as to enhance the defense and resistance of the tobacco leaves to the TMV (SEO et al, 2003). The research of Liu Yan Hua and the like shows that the tobacco mosaic disease prevention effect of the cis-abienol preparation is enhanced along with the increase of the concentration of the cis-abienol within the concentration range of 50-300 ppm. In the application of cis-abienol, only application No. 202010360951.5 proposes the application of cis-abienol in preventing and controlling crop bacterial wilt, and the bacterial wilt for preventing and controlling is that of tomato in solanaceae.
The Potato Virus Y (PVY) belongs to the genus of the potato virus Y (Poteyvirus), has a wide distribution range, is distributed in each main tobacco area in China, infects tobacco to form necrotic or mosaic symptoms, seriously influences the yield and quality of crops, and is one of main virus diseases of the tobacco. In order to prevent and cure the virus, the prior art mainly adopts a vaccine, such as application No. 201810949941.8, an attenuated vaccine for resisting the potyvirus, the attenuated vaccine is based on TVBMV attenuated mutant, and effective gene segments which can induce the potyvirus to generate cross protection are embedded in the TVBMV attenuated mutant. For another example: application No. 201910819867.2 relates to a target for screening a drug for preventing and treating potato virus Y, in particular to a drug for preventing and treating potato virus Y with potato virus Y coat protein (PVY-CP) as a target. For another example: application No. 201410362991.8A method for making tobacco obtain potato virus Y resistance, spraying the Agrobacterium tumefaciens and PBNTRA6 Agrobacterium tumefaciens in a volume ratio of 1:1, infecting the cut tobacco seedlings with agrobacterium tumefaciens, and introducing the CP gene segment into tobacco through infection.
The plant source active substances are generally insoluble in water, can not be directly applied to crops at low concentration, and need to be prepared into pesticide preparations, the forms of the preparations are called dosage forms, the pesticide preparations are different dosage forms which are distinguished by different forms and purposes of original medicine after being processed, and can be divided into powder, wettable powder, missible oil, suspending agents, emulsion in water and the like, and the dosage forms of the plant source pesticides are processed by imitating chemical pesticides for most parts. With the attention of people to the environment protection and the requirement of sustainable development, the traditional pesticide formulations such as missible oil and the like contain toxic solvents such as toluene, xylene and the like, so that the problems of easy phytotoxicity generation, environmental pollution and the like exist. The pesticide Microemulsion (ME) belongs to a thermodynamically stable single-phase system, takes water as a dispersoid, has high safety, simple processing technology, low cost and good stability over time, and becomes an important development direction of green dosage forms.
Disclosure of Invention
In order to overcome the problems that the type and the effect of the tobacco cis-abienol for preventing and treating the virus diseases are not clear in the development process of the plant source pesticide and a green formulation with commercial value is not formed, the invention researches and provides the microemulsion containing the tobacco cis-abienol, and provides the preparation method and the application of the tobacco cis-abienol antiviral microemulsion in preventing and treating the plant virus diseases.
In order to achieve the above purpose, the invention provides the following technical scheme: a microemulsion containing tobacco cis-abienol comprises the following components in percentage by weight: 2-4% of cis-fir alcohol extract, 30-40% of ethanol, 10-20% of fatty alcohol-polyoxyethylene ether, 3-7% of sodium dodecyl sulfate and the balance of water.
Further, the preparation method of the microemulsion containing the tobacco cis-abienol comprises the following steps:
step one, mixing the cis-fir alcohol extract, ethanol and fatty alcohol-polyoxyethylene ether in proportion, stirring and dissolving to prepare an oil phase;
step two, adding sodium dodecyl sulfate into water, stirring and dissolving to prepare a water phase;
and step three, slowly pouring the oil phase into the water phase under the stirring condition, and fully stirring and mixing to obtain the tobacco cis-abienol microemulsion.
Further, after mixing, stirring is continuously carried out for 30-35 min.
Further, the tobacco cis-abienol microemulsion is applied to prevention and treatment of plant virus diseases.
Further, the plant disease is potato virus Y disease.
Furthermore, the plant disease is one of tobacco mosaic virus and cucumber mosaic virus.
Furthermore, the tobacco cis-abienol microemulsion is diluted by 300-500 times and sprayed on leaf surfaces to prevent and treat virus diseases.
Furthermore, the concentration of an effective component cis-abienol in the tobacco cis-abienol microemulsion is 40-150 ppm.
Compared with the prior art, the invention has the beneficial technical effects that: the tobacco cis-fir alcohol extract is used as a raw material, only ethanol is used as a solvent, and a non-toxic or low-toxic emulsifier and a co-emulsifier are selected as auxiliaries to prepare the microemulsion, so that the process is simple, the industrial production is facilitated, and the environment is not polluted;
the tobacco cis-abienol microemulsion is used for preventing and treating potato virus Y, tobacco mosaic disease and cucumber mosaic disease, has the prevention effect of more than 2 times of amino-oligosaccharin, and enriches the types of Chinese botanical virus preparations.
Drawings
FIG. 1 is a photograph of a leaf surface tested in example 2 of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The formula of the tobacco cis-abienol microemulsion and the method for preparing the tobacco cis-abienol microemulsion are provided in the embodiment, the tobacco cis-abienol extract, ethanol and fatty alcohol-polyoxyethylene ether (AEO) are mixed according to the proportion of three sequences in the table 1, and are stirred by a magnetic bar for 6min to be dissolved, and then are continuously stirred for 20-30 min; adding sodium dodecyl sulfate into water, and stirring to dissolve; slowly adding the oil phase into the water phase under stirring, and continuously stirring for 30min after mixing to obtain 3 tobacco cis-abienol microemulsions with different proportions, as shown in Table 1 below.
TABLE 1 Nicotiana cis-abienol microemulsion formulation
| Serial number | The extract of the tobacco cis-fir alcohol is% | Ethanol% | Fatty alcohol polyoxyethylene ether% | Sodium dodecyl sulfate | Water% |
| 1 | 2% | 30% | 10% | 3% | Allowance of |
| 2 | 3% | 35% | 15% | 5% | Balance of |
| 3 | 4% | 40% | 20% | 7% | Allowance of |
The quality of the obtained tobacco cis-abienol microemulsion is detected, and the quality technical index of the 4% tobacco cis-abienol microemulsion is given in table 2.
TABLE 2, 4% tobacco cis-abienol microemulsion quality technical index
| Quality technical index | Measured value | Detection method |
| Transparent temperature range | -4℃-60℃ | Refrigerator, water bath detection |
| Microemulsion stability | Qualified | GB/T1603 |
| pH value | 7.0 | GB/T1601 |
| Low temperature stability (0 ℃. + -. 1 ℃ C. storage 10 d) | Qualified | GB/T19137, |
| Heat storage stability (storage at 50 ℃. + -. 2 ℃ C. for 10 d) | Qualified | GB/T19136 |
The prepared microemulsion meets the national standard requirements.
Example 2
The microemulsion prepared in the example 1 is taken, the influence of different dilution concentrations on tobacco leaves is tested, the microemulsion is diluted into 100, 200, 300, 400 and 500 times of liquid by water, a small amount of microemulsion is dipped by a sterilized cotton stick and is lightly rubbed on the surface of the tobacco leaves, and only epidermal cells of the leaves are required to cause micro wounds but not die.
Observing the change of the leaf surface after 48 hours, wherein 100 times of the liquid has damage to the leaf surface, and 200 times of the liquid has slight damage; 300. 400 and 500 times of the solution has no damage to the leaf surface, as shown by comparison of the pictures in figure 1. It can be seen that the micro-emulsion has an influence on the leaf surface when the dilution factor is less than 200 times.
Example 3
This example was used in a reaction greenhouse with the ability to control viral disease using the microemulsion prepared in example 1.
And (3) experimental design:
diluting the microemulsion into 150, 200, 250, 300, 350 and 400 times of solution respectively, preparing application solution with virus juice at a ratio of 1:1, and performing PVY, CMV and TMV virus disease prevention and treatment tests respectively.
Preparation of test materials:
virus strain: TMV-U1 severe strain, which is transferred to NC89 (Nicotiana tabacum var. NC89) and other tobacco seedlings for rejuvenation for 1 time 15 days before use;
CMV-IB, which is transferred to Sansheng-NN (Nicotiana tabacum var. Samsun NN) tobacco seedlings for rejuvenation once 15 days before use to prepare virus juice for standby;
PVY N the virus is transferred to K326 (Nicotiana tabacum var. K326) tobacco seedlings for rejuvenation once 15 days before use, and virus juice is prepared for standby.
Inoculation:
the test material was triclosan-NN, and when 3 to 4 true leaves of tobacco seedlings were grown and transplanted in a flowerpot with a diameter of 4 cm, 1 leaf per pot was grown to a leaf stage of 5 to 6 for testing.
Passivation test:
preparing a mixed solution (medicament) of virus juice and microemulsion diluent with the same volume, wherein a negative control solution is the virus juice and the microemulsion diluent with the same volume, and a positive control solution is the virus juice and amino-oligosaccharin (0.5 percent and 300 times of solution) with the same volume, mixing for 30min, and then inoculating the tobacco leaves. TMV used a half-leaf inoculation method: inoculating a mixed solution of virus juice and the medicament on half leaves, inoculating a negative control solution on half leaves, and counting withered spots after 48 hours; the CMV, PVY were whole-plant smeared, 10 cigarettes each were treated, repeated 3 times, and disease indices were investigated 14 and 21 days after inoculation.
Prevention experiments:
NC89 is used as a test material, the test material is sprayed once every 3 days before inoculation, the spraying is carried out 3 times in total, 10 cigarettes are treated each time, the spraying is repeated for 3 times, the inoculation is carried out after 12-24 hours of the last spraying, and the disease condition is investigated 14 days and 21 days after the inoculation.
Treatment experiments:
the test material was 100 cigarettes, virus sap was inoculated 24 hours before drug treatment, and disease incidence was investigated 14 days and 21 days after inoculation.
The statistical method comprises the following steps: the disease index is executed according to GB/T23222-2008 tobacco pest classification and investigation method.
Disease Index (DI)
In the formula: DI-disease index, Ni-number of diseased leaves (plants) at each level, i-corresponding severity grade value of disease, N-number of total investigated leaves (plants), control effect = (medicament treatment disease index-blank control disease index)/medicament treatment disease index = 100%
And (3) test results:
TABLE 3 tobacco mosaic Virus inhibition by Nicotiana cis-abienol microemulsion
TABLE 4 preventive effect of nicotiana cis-abienol microemulsion on cucumber mosaic virus
TABLE 5 prevention of Potato Virus Y by Nicotiana tabacum cis-abienol microemulsion
The results of the indoor inhibition measurement of the nicotianol microemulsion on the tobacco mosaic virus, the cucumber mosaic virus and the potato virus Y are shown in tables 3, 4 and 5, and the results show that: the inhibition rate of the 300-800 times dilution liquid on the in vitro passivation of three virus diseases ranges from 82.94% to 30.72%, and the in vitro passivation of the virus is reduced along with the reduction of the use concentration. In addition, the tobacco cis-abienol microemulsion has a protection effect on the three virus diseases by 300-800 times, the prevention effect range is 70.78-15.48%, but no treatment effect is generated.
Example 4
The field antiviral control of the tobacco cis-abienol microemulsion is carried out in 2019-2020 by Qingdao ink-test farm of tobacco institute of China academy of agricultural sciences, and PVY, CMV and TMV virus disease control tests are respectively carried out.
The test materials are 300/400/500 times of solution of Honghuadajinyuan, Nicotiana cis-abienol microemulsion and 300 times of medicament of amino-oligosaccharin. The fertilizer is applied in the tobacco plant block period, sprayed once every 10 days for 2-3 times continuously, and naturally attacks diseases in the field until the disease condition is investigated for a long time. The statistical method was the same as in example 3.
The test results are shown in table 7.
TABLE 7 field antiviral control of nicotiana cis-abienol microemulsion
And (4) conclusion: the field prevention effect of the tobacco cis-abienol microemulsion on three virus diseases reaches 2 times or more of that of the amino-oligosaccharin, and the maximum effect can reach more than 77%.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes in the embodiments and/or modifications of the invention can be made, and equivalents and modifications of some features of the invention can be made without departing from the spirit and scope of the invention.
Claims (8)
1. A microemulsion containing tobacco cis-abienol, which is characterized in that: according to the weight percentage: 2-4% of cis-fir alcohol extract, 30-40% of ethanol, 10-20% of fatty alcohol-polyoxyethylene ether, 3-7% of sodium dodecyl sulfate and the balance of water.
2. The method of preparing a microemulsion containing nicotiana cis-abienol as claimed in claim 1, wherein the steps are as follows: mixing the tobacco cis-fir alcohol extract, ethanol and fatty alcohol-polyoxyethylene ether in proportion, and stirring for dissolving to prepare an oil phase;
step two, adding sodium dodecyl sulfate into water, stirring and dissolving to prepare a water phase;
and step three, slowly pouring the oil phase into the water phase under the stirring condition, and fully stirring and mixing to obtain the tobacco cis-abienol microemulsion.
3. The method of preparing a microemulsion containing nicotiana cis-abienol as set forth in claim 2, wherein: and in the third step, after mixing, continuously stirring for 30-35 min.
4. Use of the tobacco cis-abienol microemulsion according to claim 1 for the control of plant viral diseases.
5. Use according to claim 4, characterized in that: the plant disease is potato virus Y.
6. Use according to claim 4, characterized in that: the plant disease is one of tobacco mosaic disease and cucumber mosaic disease.
7. Use according to any one of claims 4 to 6, characterized in that: the tobacco cis-abienol microemulsion is diluted by 300-500 times, and is sprayed on leaf surfaces to prevent and treat virus diseases.
8. The nicotiana cis-abienol-containing microemulsion according to claim 1, wherein: the concentration of the effective component cis-abienol is 40-150 ppm.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103120189A (en) * | 2013-02-06 | 2013-05-29 | 杨凌农科大无公害农药研究服务中心 | Cephalotaxus sinensis Li. plant-virus-resisting agent and preparation method thereof |
| CN103766414A (en) * | 2014-01-08 | 2014-05-07 | 杨凌农科大无公害农药研究服务中心 | Plant source antiviral agent containing phyllanthi fructus and preparation method thereof |
| CN111387183A (en) * | 2020-04-30 | 2020-07-10 | 中国农业科学院烟草研究所 | Application of labdanum compounds in preventing and treating crop bacterial wilt |
| CN111700078A (en) * | 2020-05-14 | 2020-09-25 | 中国农业科学院烟草研究所 | Application of firework extract in plant virus disease prevention and control |
| CN112704065A (en) * | 2020-12-30 | 2021-04-27 | 中国农业科学院烟草研究所 | Preparation method and application of tobacco cembrane diterpene microemulsion |
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- 2022-07-19 CN CN202210845060.8A patent/CN114982790A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103120189A (en) * | 2013-02-06 | 2013-05-29 | 杨凌农科大无公害农药研究服务中心 | Cephalotaxus sinensis Li. plant-virus-resisting agent and preparation method thereof |
| CN103766414A (en) * | 2014-01-08 | 2014-05-07 | 杨凌农科大无公害农药研究服务中心 | Plant source antiviral agent containing phyllanthi fructus and preparation method thereof |
| CN111387183A (en) * | 2020-04-30 | 2020-07-10 | 中国农业科学院烟草研究所 | Application of labdanum compounds in preventing and treating crop bacterial wilt |
| CN111700078A (en) * | 2020-05-14 | 2020-09-25 | 中国农业科学院烟草研究所 | Application of firework extract in plant virus disease prevention and control |
| CN112704065A (en) * | 2020-12-30 | 2021-04-27 | 中国农业科学院烟草研究所 | Preparation method and application of tobacco cembrane diterpene microemulsion |
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