CN108244103B - Use of tsaoko essential oil and its emulsion as pesticide - Google Patents
Use of tsaoko essential oil and its emulsion as pesticide Download PDFInfo
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- CN108244103B CN108244103B CN201810019834.5A CN201810019834A CN108244103B CN 108244103 B CN108244103 B CN 108244103B CN 201810019834 A CN201810019834 A CN 201810019834A CN 108244103 B CN108244103 B CN 108244103B
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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
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- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
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- A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof
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Abstract
The invention relates to the use of tsaoko amomum fruit essential oil and emulsion thereof as pesticide, which can be widely used for preventing and treating pathogenic bacteria of agriculture, forestry and animal husbandry crops. The tsaoko essential oil contains the following characteristic components in percentage by weight: (1)1, 8-cineole: 45 to 65 percent; and (2) citral: 1 to 3.5 percent. The amomum tsao-ko essential oil emulsion comprises the following components in percentage by weight: (1) 10 parts of tsaoko amomum fruit essential oil; (2) 20-35 parts of a solvent; (3) 5-15 parts of an auxiliary agent; and (4) 40-55 parts of water.
Description
Technical Field
The invention relates to a use of plant-derived tsaoko essential oil and an emulsion thereof as pesticides, which can be widely used for preventing and treating pathogenic bacteria of agriculture, forestry and animal husbandry crops.
Background
The pesticide plays an important role in agriculture, forestry and animal husbandry production, and can be used for preventing and treating pests which harm the agriculture, forestry and animal husbandry production. Currently, the commonly used pesticides are mainly chemically synthesized pesticides, wherein the bactericide pesticides mainly comprise organophosphorus, disulfide carbamate, sulfamic acid, thiosulfonate, trichloromethyl sulfide, organophosphate, organoarsenic, organotin, organosulfur, benzene, heterocycle and the like. These types of chemical germicides often have some toxicity associated with the germicidal and bacteriostatic effects.
Long-term consumption of pesticide-contaminated vegetables is an important cause of cancer, arteriosclerosis, cardiovascular diseases, fetal deformity, stillbirth, premature death, senilism and other diseases. Therefore, the security of agricultural products is very important in more and more countries in the world, and the limitation on pesticide residues is very strict because the security of agricultural products is related to the nationality. Meanwhile, the pesticide residue exceeding the standard is also a problem often encountered by import and export of agricultural products, and the competitiveness of the agricultural products in the international market is influenced. With the increase of health consciousness of people, the efforts of reducing the pesticide residue and developing novel pesticides become urgent.
Natural plants are a resource treasure house on the earth, and the human beings have already studied natural plant source substances for thousands of years and are the main sources of medicines. In recent years, with the understanding of people on the characteristics of nature, health, low toxicity, no chemical residue and the like of plant source preparations, the global market has increasingly increased demand for natural healthy raw material products, and the development of the plant extract industry is faster and faster. Therefore, in recent years, research, development and application of plant-derived preparations have been gradually regarded and greatly developed by those skilled in the art.
The botanical pesticide is a pesticide developed by utilizing plant resources, is natural, and if the plant has no toxicity, the botanical pesticide prepared by the plant does not pollute the environment and agricultural products, does not accumulate toxicity in the environment and human body, and is relatively safe to human and livestock. Moreover, the plant-derived pesticide is not a single compound, is all or a part of organic substances of plant organisms, has a complex composition, is difficult to generate resistance to pests or harmful microorganisms, and has the characteristics of low toxicity and low residue, so the plant-derived pesticide has a wide market prospect.
At present, the development of botanical bacteriostatic pesticide is mainly focused on the families of Compositae, Leguminosae, Umbelliferae, Gramineae, Labiatae, Magnoliaceae, Aristolochiaceae, Polygonaceae, Oleaceae, Liliaceae, Cucurbitaceae, Cyperaceae, Brassicaceae, Lauraceae and the like. Wherein the substance with antibacterial activity is terpenes, alkaloids, phenols, polyacetylenes, unique polypeptide and polysaccharide, etc.
Tsaoko (Amomum tsaoko) is a perennial herb of Zingiberaceae (Zingifera) and Amomum (Amomum), and is mainly distributed in provinces of Yunnan, Guangxi, Guizhou, etc. in China. The tsaoko amomum fruit has special and strong spicy flavor, is mainly used as a flavoring and is an excellent product in cooking seasonings. At present, there are few reports of plant extracts obtained from plants of the family zingiberaceae as botanical pesticides.
Disclosure of Invention
The present invention relates to the use of essential oil of tsaoko amomum and its emulsion extracted from the fruit of dry ripe tsaoko amomum as a botanical pesticide.
In a first aspect, the present invention provides the use of a tsaoko essential oil as a plant-derived pesticide, said tsaoko essential oil containing the following characteristic components by weight:
(1)1, 8-cineole: 45 to 65 percent;
(2) alpha-pinene: 2 to 4 percent;
(3) geraniol: 0 to 4 percent; and
(4) geranial: 5 to 8 percent.
Preferably, the content of 1, 8-cineole is 50-63% by weight. Preferably, the content of 1, 8-cineole is 58-63% by weight.
In a second aspect, the present invention provides the use of an essential oil of tsaoko fruit as a plant source, prepared by a method comprising:
(1) weighing a certain amount of dried tsaoko amomum fruits, and knocking into small pieces or crushing into coarse powder;
(2) extracting tsaoko essential oil from small pieces or coarse powder of dried tsaoko; and
(3) and collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield.
The method for extracting the amomum tsao-ko essential oil is selected from: steam distillation method, solvent extraction method, CO2Supercritical extraction technology, subcritical water extraction and microwave-assisted extraction.
In the extraction method using a steam distillation method:
the weight ratio of the dried small pieces or coarse powder of the tsaoko amomum fruit to the water is 1:3-1:20, and the dried small pieces or coarse powder of the tsaoko amomum fruit are soaked in the water for 0-24 hours; then distilling and extracting for 0.5-10 hours by steam; and collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield.
Preferably, the dried small tsaoko pieces or coarse powder are soaked in water for 0-4 hours in a weight ratio of the dried small tsaoko pieces or coarse powder to water of 1:3-1: 10; then distilling and extracting for 3-8 hours by steam; and collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield.
Preferably, the weight ratio of the dried small pieces or coarse powder of the tsaoko amomum fruits to the water is 1:4, and the dried small pieces or coarse powder of the tsaoko amomum fruits are soaked in the water for 1 hour; then distilling and extracting for 7 hours by steam; and collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield.
The dried tsaoko can be dried tsaoko obtained by air-drying or dried tsaoko obtained by oven-drying.
The oil yield of the amomum tsao-ko essential oil of the invention is in the range of 1.0-2.5% by weight. The tsaoko amomum essential oil is a light yellow transparent oily substance, has special strong fragrance and abnormal mellow fragrance, and is refreshing.
The tsaoko essential oil of the invention is used as an active ingredient of pesticide and can be prepared into solid, liquid, emulsion, spraying agent and aerosol. For example, the liquid may be used as it is, or may be prepared into a solid preparation by a process such as embedding. In the case of sprays, the propellant used is selected from the group consisting of chlorofluorocarbons, hydrocarbons and compressed gas.
The pesticide with the amomum tsao-ko essential oil as an active ingredient can be used for preventing and treating plant diseases caused by the following plant pathogenic bacteria: aspergillus flavus, Cladosporium cladosporioides, Penicillium commune, Penicillium chrysogenum, Rhizoctonia solani, Botrytis cinerea, Alternaria ginseng (Alternaria panax), Fusarium solani (Fusarium solani), Fusarium oxysporum (Fusarium oxysporum), Rhizoctonia solani (Fusarium oxysporum), Rhizoctonia pseudochinensis (Ilyocytria mors-panicis) and/or Haemophilus robusta (Ilyocytria robusta).
Specifically, the pesticide using the amomum tsao-ko essential oil of the invention as an active ingredient can be used for preventing and treating the following plant diseases: medicinal plants selected from Ginseng radix, radix Panacis Quinquefolii, Notoginseng radix, Bulbus Fritillariae Cirrhosae, radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, Atractylodis rhizoma, Bulbus Lilii, radix Rhodiolae, peony and Aloe; a field crop selected from wheat, corn, rice, peanut and soybean; vegetable crops selected from potato, tomato, cucumber, celery, pepper, eggplant, onion; selected from commercial crops such as tobacco; a fruit crop selected from grapes, citrus, strawberry, apple, banana, loquat and passion flower.
Specifically, the pesticide using the amomum tsao-ko essential oil of the invention as an active ingredient can be used for preventing and treating the following plant diseases:
damping off, gray mold, black spot and root rot of ginseng, American ginseng and pseudo-ginseng; gray mold and bulb rot of fritillary bulb; radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, radix Rhodiolae, Atractylodis rhizoma, peony and Aloe root rot; lily wilt; wheat basal stem rot and root rot; ear rot and grain rot of corn; rice ear rot; peanut root rot and rot; soybean root rot; potato dry rot; tomato gray mold, damping off, blight and leaf spot; gray mold, damping off and root rot of cucumber; root rot and damping off of celery, pepper and eggplant; onion penicilliosis; tobacco root rot; grape gray mold and black root disease; penicillium citrinum; gray mold and root rot of strawberry; apple mildew heart disease and gray mold; banana wilt; loquat rot disease; passion fruit rot.
In a third aspect, the present invention provides the use of an emulsion of tsaoko essential oil as a botanical pesticide, the emulsion of tsaoko essential oil having the following composition by weight:
(1) 10 parts of tsaoko amomum fruit essential oil;
(2) 20-35 parts of a solvent;
(3) 5-15 parts of an auxiliary agent; and
(4) 40-55 parts of water.
The solvent is selected from: methanol, ethanol, N-butanol, N-propanol, isopropanol, isobutanol, N-pentanol, isopentanol, N-hexanol, acetone, dichloromethane, ethyl acetate, dimethyl sulfoxide, tetrahydrofuran, dimethylformamide, cyclohexanone, pyrrolidone, N-methylpyrrolidone, or a mixture of two or more thereof. Preferably, the solvent is selected from: ethanol, methanol, propanol, isopropanol, butanol, glycerol, acetone, etc., or a mixture of two or more thereof. Preferably, the solvent is methanol, ethanol or a mixture of both.
The auxiliary agent is a surfactant, a carrier and/or a dispersing agent, the surfactant is selected from tweens, spans, sodium dodecyl sulfate, alcohols, propylene glycol, polyethylene glycol and glycerol, the dispersing agent is selected from sodium hexametaphosphate, sodium pyrophosphate, sodium polyacrylate and polyvinyl alcohol, and the carrier is selected from liposomes, cyclodextrin and microcapsules. Preferably, the surfactant is selected from the group consisting of tweens, alkylphenol polyoxyethylene ethers, spans, sodium lauryl sulfate. Preferably, the surfactant is selected from the group consisting of tweens, alkylphenol ethoxylates.
The emulsion of the amomum tsao-ko essential oil of the invention is selected from:
1 part of tsaoko essential oil, 3.5 parts of ethanol, 1.5 parts of tween surfactant and 4 parts of water, and uniformly stirring to obtain the composition;
1 part of tsaoko essential oil, 2 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene ether surfactant and 5.5 parts of water, and uniformly stirring to obtain the product;
1 part of tsaoko essential oil, 3.5 parts of ethanol, 0.5 part of alkylphenol polyoxyethylene ether surfactant and 4 parts of water, and uniformly stirring to obtain the product;
1 part of tsaoko essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene ether surfactant and 4 parts of water, and uniformly stirring to obtain the product; and
1 part of tsaoko essential oil, 2.5 parts of ethanol, 1 part of methanol, 1.5 parts of alkylphenol polyoxyethylene ether surfactant and 4 parts of water, and uniformly stirring to obtain the composition.
The pesticide containing the amomum tsaoko essential oil emulsion of the invention as an active ingredient can be applied to the following objects: medicinal plants selected from Ginseng radix, radix Panacis Quinquefolii, Notoginseng radix, Bulbus Fritillariae Cirrhosae, radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, Atractylodis rhizoma, Bulbus Lilii, radix Rhodiolae, peony and Aloe; a field crop selected from wheat, corn, rice, peanut and soybean; vegetable crops selected from potato, tomato, cucumber, celery, pepper, eggplant, onion; selected from commercial crops such as tobacco; a fruit crop selected from grapes, citrus, strawberry, apple, banana, loquat and passion flower.
The pesticide with the amomum tsao-ko essential oil emulsion as an active ingredient can be used for preventing and treating plant diseases caused by the following plant pathogenic bacteria: aspergillus flavus, Cladosporium cladosporioides, Penicillium commune, Penicillium chrysogenum, Rhizoctonia solani, Botrytis cinerea, Alternaria ginseng (Alternaria panax), Fusarium solani (Fusarium solani), Fusarium oxysporum (Fusarium oxysporum), Rhizoctonia solani (Fusarium oxysporum), Rhizoctonia pseudochinensis (Ilyocytria mors-panicis) and/or Haemophilus robusta (Ilyocytria robusta). The tsaoko amomum essential oil emulsion has an inhibiting effect on all pathogenic fungi, and particularly has an excellent inhibiting effect on fungi which are difficult to control, such as fusarium and cuprum.
Specifically, the pesticide using the amomum tsaoko essential oil emulsion of the invention as an active ingredient can be used for preventing and treating the following plant diseases:
damping off, gray mold, black spot and root rot of ginseng, American ginseng and pseudo-ginseng; gray mold and bulb rot of fritillary bulb; radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, radix Rhodiolae, Atractylodis rhizoma, peony and Aloe root rot; lily wilt; wheat basal stem rot and root rot; ear rot and grain rot of corn; rice ear rot; peanut root rot and rot; soybean root rot; potato dry rot; tomato gray mold, damping off, blight and leaf spot; gray mold, damping off and root rot of cucumber; root rot and damping off of celery, pepper and eggplant; onion penicilliosis; tobacco root rot; grape gray mold and black root disease; penicillium citrinum; gray mold and root rot of strawberry; apple mildew heart disease and gray mold; banana wilt; loquat rot disease; passion fruit rot.
The plant diseases caused by the fungi can be specifically as follows:
1. and (3) aspergillus flavus: corn ear rot, grain rot, and peanut rot;
2. blue mold: citrus penicillium, onion penicillium;
3. producing penicillium chrysogenum: penicillium citrinum;
4. cladosporium dendritic: tomato leaf spot, apple core rot;
5. fusarium solani: ginseng, American ginseng, pseudo-ginseng, astragalus, lily, peony, liquorice, platycodon grandiflorum, salvia miltiorrhiza, safflower, aloe root rot and fritillaria bulb rot; wheat stem and root rot, soybean root rot; pepper, eggplant, cucumber, sweet potato, peanut, celery root rot, tomato wilt and root rot, potato wilt; tobacco root rot; root rot of watermelon, orange and strawberry;
6. fusarium oxysporum including radix Ginseng, radix Panacis Quinquefolii, radix Notoginseng, rhizoma Chuanxiong, radix astragali, radix Glycyrrhizae, radix Rhodiolae, radix Ophiopogonis, rhizoma Bletillae, rhizoma Atractylodis Macrocephalae, flos Farfarae, radix Isatidis root rot, and Bulbus Lilii wilt; wheat stem and root rot, rice ear rot; tomato fusarium wilt and root rot, cucumber fusarium wilt and root rot, onion dry rot, potato dry rot; tobacco root rot; strawberry root rot, banana wilt;
7. pseudo-ginseng and red husks: red skin disease of ginseng, root rot of American ginseng;
8. strengthening soil and red husk: ginseng red skin disease, root rot of American ginseng and pseudo-ginseng, and grape black root disease;
9. botrytis cinerea: ginseng, American ginseng, pseudo-ginseng, fritillaria, apple, tomato, grape, strawberry gray mold, loquat rot, passion fruit rot;
10. rhizoctonia solani: ginseng, American ginseng, pseudo-ginseng, celery, hot pepper, eggplant and tomato damping off;
11. ginseng alternaria alternate: black spot of ginseng, American ginseng and notoginseng.
Description of the drawings:
FIG. 1 shows a GC-MS chromatogram of the tsaoko essential oil of the present invention.
Figure 2 is a graph showing the comparison of the inhibitory effect of the tsaoko essential oil emulsion on 6 plant pathogens.
Figure 3 illustrates the control effect of the tsaoko essential oil emulsion on Fusarium sp.
Detailed Description
The technical solution of the present invention is further described below with reference to the following embodiments and the accompanying drawings. These examples are given solely for the purpose of illustration and are not intended to limit the scope of the invention.
Example 1: preparation of tsaoko essential oil
Weighing 0.5kg of each of the air-dried and oven-dried fructus Tsaoko, pulverizing into coarse powder, and soaking in 3.5L water for 1 h. Steam distilling for 7h, collecting fructus Tsaoko essential oil with volatile oil extractor, and calculating oil yield (oil yield/weight of fructus Tsaoko) x 100%, wherein the oil yield of air-dried fructus Tsaoko is 2.0%, and the oil yield of oven-dried fructus Tsaoko is 1.3%.
Example 2: characterization of Amomum tsaoko essential oil
Analysis of the composition of essential oil of Amomum tsao-ko
The components and contents of the essential oil of the tsaoko amomum fruits extracted from the air-dried and oven-dried tsaoko amomum fruits are analyzed by a GC/MS method. Diluting the fructus Tsaoko essential oil with n-hexane by 10000 times, and injecting 1 μ l. GC conditions were as follows: a Thermo TR-5MS (30m x 0.32mm, 0.25 μm) chromatographic column is adopted, the initial temperature of the column is 50 ℃, the temperature is programmed to 6 ℃/min and is increased to 220 ℃, then the temperature is increased to 250 ℃ at 10 ℃/min, and the constant temperature is kept for 5 min. The injection port temperature is 250 ℃, the carrier gas He, the carrier gas flow rate is 2.0ml/min, and the split ratio is 20: 1. MS conditions: EI ion source, scanning mode full scan, scanning range m/z 50-600amu, ion source temperature 250 ℃ and GC/MS interface temperature 250 ℃.
The GC-MS chromatogram of the tsaoko essential oil is shown in figure 1. The results show that 29 chromatographic peaks are detected in the amomum tsao-ko essential oil extracted from the air-dried amomum tsao-ko, wherein 25 volatile matters are identified, which account for 98.7 percent of the total relative content of the essential oil, wherein the relative content of the 1, 8-cineole is 58.11 percent, and the relative content of the citral is 3.33 percent. The first sample of the amomum tsao-ko essential oil extracted from the roasted amomum tsao-ko was co-detected with 33 peaks, of which 30 volatiles were identified, accounting for 97.75% of the total relative content of the essential oil, of which the relative content of 1, 8-cineole is 62.73% and the relative content of citral is 2.92%. The second sample of oven-dried tsaoko was examined for 32 peaks of spectrum for the tsaoko essential oil, of which 30 volatiles were identified, accounting for 97.12% of the total relative content of the essential oils, with a relative content of 1, 8-eucalyptol of 50.29% and a relative content of citral of 3.34%.
The specific components and contents are shown in the following table 1:
table 1 comparison of chemical composition of tsaoko essential oil extracted from air-dried and oven-dried tsaoko:
as can be seen from the above table, the three tsaoko essential oils obtained from the three tsaoko samples had the following two main components and contents:
1, 8-cineole: 50 to 63 percent of
Citral: 2.9-3.4 percent.
Evaluation of odor of tsaoko essential oil: special and strong spicy flavor, abnormal mellow flavor and refreshing mind and spleen.
Example 3: preparation of tsaoko essential oil emulsion
1 part of tsaoko essential oil, 3.5 parts of ethanol, 801.5 parts of tween and 4 parts of water, and uniformly stirring to obtain the composition.
1 part of tsaoko amomum fruit essential oil, 2 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 5.5 parts of water, and uniformly stirring to obtain the composition.
1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 0.5 part of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
1 part of tsaoko amomum fruit essential oil, 2.5 parts of ethanol, 1 part of methanol, 1.5 parts of alkylphenol polyoxyethylene ether (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
Evaluation of odor of tsaoko essential oil emulsion: special and strong spicy flavor, abnormal mellow flavor and refreshing mind and spleen.
Example 4: amomum tsao-ko essential oil and Amomum tsao-ko essential oil emulsion fungus inhibition test
Experimental strains: aspergillus flavus (Aspergillus flavus) and Cladosporium cladosporioides (Cladosporium cladosporioides).
And (3) amomum tsao-ko essential oil emulsion: the following emulsion formulation of example 3 was used:
1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
Toxicity test- -hypha growth rate method
1) Adding the tsaoko essential oil into the culture medium according to the dilution multiple of 5000 times, uniformly mixing, cooling, similarly, adding the tsaoko essential oil emulsion into the culture medium according to the dilution multiple of 500 times (wherein the concentration of the tsaoko essential oil in the tsaoko essential oil emulsion is the same as that of the tsaoko essential oil diluted by 5000 times), uniformly mixing, and cooling;
2) adding the tsaoko essential oil into the culture medium according to the dilution multiple of 4000 times, uniformly mixing, cooling, similarly, adding the tsaoko essential oil emulsion into the culture medium according to the dilution multiple of 500 times (wherein the concentration of the tsaoko essential oil in the tsaoko essential oil emulsion is lower than that of the tsaoko essential oil diluted by 4000 times), uniformly mixing, and cooling;
3) inoculating aspergillus flavus in the culture medium of the step 1), inoculating cladosporium cladosporioides in the culture medium of the step 2), inoculating 10 mu L of spore suspension into each plate, and repeating 6 times to serve as a test sample group;
3) the plates without the medicament were inoculated with 3 strains, each plate was inoculated with 10 μ L spore suspension, 6 replicates were used as blank control;
4) the cells were cultured in an incubator at 25 ℃ for 7 days.
And (4) judging and stipulating:
the colony diameter (mm) was measured, and the inhibition ratio was calculated as ((control group colony diameter-test group colony diameter)/control group colony diameter) × 100%.
TABLE 2 comparison of the antifungal Effect of the Amomum tsao-ko essential oil and the Amomum tsao-ko essential oil emulsion
From the results in the table 2, it can be seen that the inhibition rate of the tsaoko amomum fruit essential oil on aspergillus flavus is 39.8% when the tsaoko amomum fruit essential oil is diluted by 5000 times, the same concentration is achieved when the emulsion containing 10% of the tsaoko amomum fruit essential oil is diluted by 500 times, the inhibition rate on aspergillus flavus is up to 72.2%, and the bacteriostasis effect is superior to that of the tsaoko amomum fruit essential oil. The inhibition rate of the amomum tsao-ko essential oil to the cladosporium cladosporioides is 70.19 percent when the amomum tsao-ko essential oil is diluted by 4000 times, the concentration of the emulsion containing 10 percent of the amomum tsao-ko essential oil is lower than that when the amomum tsao-ko essential oil is diluted by 500 times, the inhibition rate to the cladosporium cladosporioides is as high as 100 percent, and the bacteriostasis effect is better than that of the amomum tsao-ko essential oil.
Example 5: bacteriostatic action of tsaoko essential oil emulsion on fungi
Experimental strains:
penicillium polonicum (Penicillium polonicum).
Culture medium: potato agar culture medium
The formula of the tsaoko essential oil emulsion is as follows:
1 part of tsaoko essential oil, 3.5 parts of ethanol, 801.5 parts of tween and 4 parts of water, and uniformly stirring to obtain the composition.
Virulence determination test- -EC50(half maximal effect concentration):
1) preparing a suspension of the blue penicillium, wherein the concentration of the bacterial liquid is 108cfu/mL；
2) Adding the tsaoko amomum essential oil emulsion into a PDA culture medium at the concentration gradient of 500, 200, 100, 50 and 25 mu L/L, uniformly mixing and cooling;
3) inoculating Penicillium poluense at each concentration, inoculating 10 μ L of each plate, and repeating 6 times to obtain test sample groups;
4) respectively inoculating penicillium poluense to the plates without the amomum tsao-ko essential oil preparation, wherein each plate is inoculated with 10 mu L of penicillium poluense, and 6 times of the plates are used as blank control groups;
5) the cells were cultured in an incubator at 25 ℃ for 7 days.
And (4) judging and stipulating:
the colony diameter (mm) was measured, and the inhibition ratio was calculated as ((control group colony diameter-test group colony diameter)/control group colony diameter) × 100%. The inhibition ratio was converted into a several-degree value (Y), and the drug mass concentration (μ L/L) was converted into a paired value (X), and a regression equation of virulence (Y ═ a + bX) andcorrelation coefficient (R), calculating the concentration of the agent (EC) that inhibits the growth of the test strain by 50%50)。
The bacteriostatic results of the tsaoko essential oil emulsion on penicillium pullulans are shown in table 3 below.
TABLE 3 bacteriostatic effect of tsaoko amomum fruit essential oil emulsion bacteriostatic agent on penicillium polish
Example 6: bacteriostatic action of tsaoko essential oil emulsion on fungi
Experimental strains:
aspergillus flavus (Aspergillus flavus), Cladosporium cladosporides (Cladosporium cladosporioides) and Penicillium chrysogenum (Penicillium chrysogenum).
Culture medium: potato agar culture medium
The formula of the tsaoko essential oil emulsion is as follows:
1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
Toxicity test- -hypha growth rate method
1) Adding the tsaoko amomum essential oil emulsion into a culture medium according to the dilution times of 500 times and 1000 times, uniformly mixing, and cooling;
2) inoculating 3 strains at each concentration, inoculating 10 mu L of each plate, and repeating 6 strains to serve as a test sample group;
3) the plates without the drug were inoculated with 3 strains, 10 μ L of each plate, 6 replicates, as a blank control;
4) the cells were cultured in an incubator at 25 ℃ for 7 days.
And (4) judging and stipulating:
the colony diameter (mm) was measured, and the inhibition ratio was calculated as ((control group colony diameter-test group colony diameter)/control group colony diameter) × 100%.
The experimental results show (table 4) that the tsaoko amomum fruit essential oil emulsion has strong inhibition effect on plant pathogenic bacteria of Aspergillus flavus (Aspergillus flavus), Cladosporium cladosporioides (cladosporides) and Penicillium chrysogenum (Penicillium chrysogenum).
TABLE 4 bacteriostatic effect of tsaoko essential oil emulsion on fungi
The results of examples 4-6 show that the tsaoko essential oil and tsaoko essential oil emulsion of the present invention exhibit bacteriostatic effects on the phytopathogenic fungi aspergillus flavus, cladosporium cladosporioides, penicillium polans and penicillium chrysogenum.
Example 7: inhibition effect of tsaoko amomum fruit essential oil emulsion on 7 plant pathogenic bacteria
Experimental strains: rhizoctonia solani, Botrytis cinerea, Alternaria ginseng (Alternaria panax), Fusarium solani, Fusarium oxysporum (Fusarium oxysporum), Pseudocercus elatus (Ilyonectria moore-panax), and Pseudocercus robusta (Ilyonectria robusta) isolated from diseased Panax quinquefolium or Panax ginseng.
And (3) amomum tsao-ko essential oil emulsion: an emulsion of the following formulation in example 3 was used:
1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
Culture medium: potato agar culture medium
Toxicity test- -hypha growth rate method
1) Beating the pathogenic bacteria cultured for 7 days into a fungus cake with the diameter of 6 mm;
2) adding the tsaoko amomum essential oil emulsion into a culture medium according to the dilution multiple of 500 times, uniformly mixing, and cooling;
3) each plate was inoculated with 1 cake at 5 replicates per concentration as a test sample set;
4) plates without the agent were inoculated with 7 strains, 5 replicates, respectively, as blank controls;
5) the cells were cultured in an incubator at 25 ℃ for 5 days.
Rules for judgment
The diameter (mm) of the bacterial colony is measured, and the bacteriostasis rate is calculated, wherein the bacteriostasis rate is (the diameter of the control group bacterial colony-the diameter of the test group bacterial colony)/(the diameter of the control group bacterial colony-the diameter of the bacterial cake) multiplied by 100 percent.
The results of the experiment are shown in table 5 below:
table 5 bacteriostatic effect of the amomum tsao-ko essential oil emulsion on 7 plant pathogens:
# denotes that the control colony diameter reached 9cm before this time point.
The table 5 shows that the tsaoko amomum fruit essential oil emulsion has an inhibiting effect on 7 pathogenic fungi, and particularly has an excellent bacteriostatic effect on fungi which are difficult to control, such as fusarium and cuprum.
Example 8: prevention and treatment effect of tsaoko essential oil emulsion on root rot of American ginseng
The formula of the tsaoko essential oil emulsion is as follows: 1 part of tsaoko amomum fruit essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene (OP10) and 4 parts of water, and uniformly stirring to obtain the composition.
Sample preparation:
healthy 4-year-old American ginseng root.
Evaluation of bacteriostatic Effect
1) Sterilizing the surface of American ginseng root, placing in a culture dish paved with 2 layers of sterilized filter paper, preparing Fusarium sp spore suspension, and soaking American ginseng root in the spore suspension for 2min for inoculation;
2) 1ml of the American ginseng root after the tsaoko amomum fruit essential oil emulsion is sprayed and inoculated is used as a medicament treatment sample, 1ml of sterile water is used for replacing the tsaoko amomum fruit essential oil emulsion as a control, and each group is provided with 3 American ginseng roots;
3) 1ml of sterile water was added to each dish, and the mixture was cultured in an incubator at 25 ℃ for 20 days. According to the disease grading standard of the American ginseng root disease in the following table 6, the disease index is calculated every 10 days, and the formula is as follows:
the disease index is 100 × Σ (the number of disease in each stage × the disease level)/(the total number × the highest value of disease level) × 100%;
the preventing and treating effect (%) is (contrast disease index-treatment disease index)/contrast disease index multiplied by 100 percent
Table 6 disease grading criteria:
the experimental results show that (table 7), after the tsaoko essential oil emulsion is subjected to spray treatment for 10 days, the prevention effect on the American ginseng root rot caused by Fusarium sp is 75%, and after 20 days, the prevention effect reaches 80%, which shows that the tsaoko essential oil emulsion has a better prevention effect on the American ginseng root rot.
Table 7 control of Fusarium (Fusarium sp.) infestation on the roots of panax quinquefolius by tsaoko essential oil emulsion:
Claims (8)
1. the application of the tsaoko essential oil emulsion as a botanical pesticide comprises the following steps:
an emulsion is obtained by uniformly stirring 1 part of tsaoko essential oil, 3.5 parts of ethanol, 1.5 parts of alkylphenol polyoxyethylene ether surfactant and 4 parts of water;
the tsaoko essential oil contains the following characteristic components in percentage by weight:
(1)1, 8-cineole: 50-63%;
(2) citral: 2.9-3.4 percent.
2. The amomum tsao-ko essential oil emulsion according to claim 1, having a content of 1, 8-cineole of 58-63% by weight.
3. Use according to claim 1 or 2, the preparation method of the tsaoko essential oil comprising:
(1) weighing a certain amount of dried tsaoko amomum fruits, and knocking into small pieces or crushing into coarse powder;
(2) extracting tsaoko essential oil from small pieces or coarse powder of dried tsaoko; and
(3) collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield;
the method for extracting the amomum tsao-ko essential oil is a steam distillation method, and the extraction method adopting the steam distillation method comprises the following steps:
soaking the dried small pieces or coarse powder of fructus Tsaoko in water for 0-4 hr at a weight ratio of 1:4-1: 10; then distilling and extracting for 3-8 hours by steam; collecting the amomum tsao-ko essential oil by using a volatile oil extractor, and calculating the oil yield; the dried tsaoko can be dried tsaoko obtained by air drying, or dried tsaoko obtained by oven drying,
the dried tsaoko can be dried tsaoko obtained by air drying or dried tsaoko obtained by oven drying.
4. Use according to claim 3, in an extraction process using steam distillation:
soaking the dried small tsaoko pieces or coarse powder in water for 1 hour, wherein the weight ratio of the dried small tsaoko pieces or coarse powder to the water is 1: 7; then distilling and extracting for 7 hours by steam; collecting fructus Tsaoko essential oil with volatile oil extractor, and calculating oil yield within 1.0-2.5 wt%,
the dried tsaoko can be dried tsaoko obtained by air drying or dried tsaoko obtained by oven drying.
5. Use according to any one of claims 1 to 4, wherein the plant-derived pesticide is useful for controlling plant diseases caused by a phytopathogen selected from the group consisting of: penicillium genus (Penicillium) Aspergillus (a), (b) and (c)Aspergillus) Sickle belongs toFusarium) Genus Geranium (A. sup.), (B. sup.), (C. sup.), (A. sup.), (B. sup.), (C. sup.), (A. sup.), (B. sup.), (C. sup., (B. sup.), (C. sup.), (B)Ilyonectria) Alternaria, Alternaria (Alternaria) Cladosporium species (A), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C)Cladosporium) Rhizoctonia solani core (Rhizoctoniasolani) And/or Botrytis cinerea (A), (B), (C)Botrytis cinerea）。
6. The use according to claim 5, wherein the pathogenic bacterium of Aspergillus is Aspergillus flavus (Aspergillus flavus) ((Aspergillus flavus))Aspergillusflavus) The pathogenic bacteria of the genus Penicillium are Penicillium Polianioides (Penicilliumpolonicum) And Penicillium chrysogenum (Penicilliumchrysogenum) The pathogenic bacteria of the fusarium are fusarium solani (f)Fusariumsolani) And Fusarium oxysporum (f.sp.) (Fusariumoxysporum) The pathogenic bacteria of the genus erythrocuprum are pseudoginseng erythrocuprum (Ilyonectriamors- panacis) And strengthening the root of Ottelia (Haw et Zhang)Ilyonectriarobusta) The pathogenic bacteria of Alternaria is Alternaria ginseng (A)Alternariapanax) The pathogenic bacteria of Cladosporium are Cladosporium cladosporioides (A), (B), (C) and (C) a)Cladosporiumcladosporioides）。
7. The use according to any one of claims 1 to 4, wherein the plant-derived pesticide is capable of being applied to a subject having: medicinal plants selected from Ginseng radix, radix Panacis Quinquefolii, Notoginseng radix, Bulbus Fritillariae Cirrhosae, radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, Atractylodis rhizoma, Bulbus Lilii, radix Rhodiolae, peony and Aloe; a field crop selected from wheat, corn, rice, peanut and soybean; vegetable crops selected from potato, tomato, cucumber, celery, pepper, eggplant, onion; selected from commercial crops such as tobacco; a fruit crop selected from grapes, citrus, strawberry, apple, banana, loquat and passion flower.
8. Use according to any one of claims 1 to 4, wherein the plant-derived pesticide is useful for controlling plant diseases caused by the following phytopathogens: damping off, gray mold, black spot and root rot of ginseng, American ginseng and pseudo-ginseng; gray mold and bulb rot of fritillary bulb; radix astragali, Glycyrrhrizae radix, radix Platycodi, flos Farfarae, radix Isatidis, Saviae Miltiorrhizae radix, Carthami flos, radix Ophiopogonis, rhizoma Bletillae, rhizoma Ligustici Chuanxiong, radix Rhodiolae, Atractylodis rhizoma, peony and Aloe root rot; lily wilt; wheat basal stem rot and root rot; ear rot and grain rot of corn; rice ear rot; peanut root rot and rot; soybean root rot; potato dry rot; tomato gray mold, damping off, blight and leaf spot; gray mold, damping off and root rot of cucumber; root rot and damping off of celery, pepper and eggplant; onion penicilliosis; tobacco root rot; grape gray mold and black root disease; penicillium citrinum; gray mold and root rot of strawberry; apple mildew heart disease and gray mold; banana wilt and loquat rot; passion fruit rot.
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| CN114532372B (en) * | 2022-02-18 | 2022-08-12 | 华南农业大学 | Application of amomum tsao-ko and waste thereof in preparation of plant source herbicide |
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