CN116965407B - Application of artemisia rupestris extract in preparation of preparation for preventing and treating tobacco black shank - Google Patents
Application of artemisia rupestris extract in preparation of preparation for preventing and treating tobacco black shank Download PDFInfo
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- CN116965407B CN116965407B CN202311217689.9A CN202311217689A CN116965407B CN 116965407 B CN116965407 B CN 116965407B CN 202311217689 A CN202311217689 A CN 202311217689A CN 116965407 B CN116965407 B CN 116965407B
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- black shank
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- 241001670235 Artemisia rupestris Species 0.000 title claims abstract description 61
- 235000015784 Artemisia rupestris Nutrition 0.000 title claims abstract description 61
- 239000000284 extract Substances 0.000 title claims abstract description 61
- 241000208125 Nicotiana Species 0.000 title claims abstract description 41
- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 41
- 241000233647 Phytophthora nicotianae var. parasitica Species 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000010992 reflux Methods 0.000 claims abstract description 5
- NPNUFJAVOOONJE-ZIAGYGMSSA-N β-(E)-Caryophyllene Chemical compound C1CC(C)=CCCC(=C)[C@H]2CC(C)(C)[C@@H]21 NPNUFJAVOOONJE-ZIAGYGMSSA-N 0.000 claims description 22
- NVEQFIOZRFFVFW-UHFFFAOYSA-N 9-epi-beta-caryophyllene oxide Natural products C=C1CCC2OC2(C)CCC2C(C)(C)CC21 NVEQFIOZRFFVFW-UHFFFAOYSA-N 0.000 claims description 11
- FAMPSKZZVDUYOS-UHFFFAOYSA-N alpha-Caryophyllene Natural products CC1=CCC(C)(C)C=CCC(C)=CCC1 FAMPSKZZVDUYOS-UHFFFAOYSA-N 0.000 claims description 11
- NPNUFJAVOOONJE-UHFFFAOYSA-N beta-cariophyllene Natural products C1CC(C)=CCCC(=C)C2CC(C)(C)C21 NPNUFJAVOOONJE-UHFFFAOYSA-N 0.000 claims description 11
- 229940117948 caryophyllene Drugs 0.000 claims description 11
- NPNUFJAVOOONJE-UONOGXRCSA-N caryophyllene Natural products C1CC(C)=CCCC(=C)[C@@H]2CC(C)(C)[C@@H]21 NPNUFJAVOOONJE-UONOGXRCSA-N 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 8
- 235000003261 Artemisia vulgaris Nutrition 0.000 claims description 6
- 238000007796 conventional method Methods 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 235000003826 Artemisia Nutrition 0.000 claims description 5
- 235000009052 artemisia Nutrition 0.000 claims description 5
- 239000000469 ethanolic extract Substances 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 238000007605 air drying Methods 0.000 claims description 3
- 244000030166 artemisia Species 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 15
- 238000000605 extraction Methods 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 231100000053 low toxicity Toxicity 0.000 abstract description 2
- 239000000419 plant extract Substances 0.000 abstract description 2
- 241000233645 Phytophthora nicotianae Species 0.000 description 26
- 230000000844 anti-bacterial effect Effects 0.000 description 25
- 230000000694 effects Effects 0.000 description 20
- 230000005764 inhibitory process Effects 0.000 description 15
- 230000012010 growth Effects 0.000 description 14
- 241000196324 Embryophyta Species 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 239000004480 active ingredient Substances 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 5
- 239000003208 petroleum Substances 0.000 description 5
- GHNBEBDYYSVNEK-XPQLPGEHSA-N Capillone Chemical compound C\C=C\C=C\C(=O)C1=CC=CC=C1 GHNBEBDYYSVNEK-XPQLPGEHSA-N 0.000 description 4
- RAZOKRUZEQERLH-UHFFFAOYSA-N Capillone Natural products CC#CC#CC(=O)C1=CC=CC=C1 RAZOKRUZEQERLH-UHFFFAOYSA-N 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 240000006891 Artemisia vulgaris Species 0.000 description 3
- 101800004637 Communis Proteins 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000001018 virulence Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- VUSBHGLIAQXBSW-UHFFFAOYSA-N 2,6-dimethyloct-7-en-4-one Chemical compound CC(C)CC(=O)CC(C)C=C VUSBHGLIAQXBSW-UHFFFAOYSA-N 0.000 description 2
- 235000001405 Artemisia annua Nutrition 0.000 description 2
- 240000000011 Artemisia annua Species 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000195474 Sargassum Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000010413 mother solution Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
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- 238000010898 silica gel chromatography Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
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- 230000001988 toxicity Effects 0.000 description 2
- KGRVJHAUYBGFFP-UHFFFAOYSA-N 2,2'-Methylenebis(4-methyl-6-tert-butylphenol) Chemical compound CC(C)(C)C1=CC(C)=CC(CC=2C(=C(C=C(C)C=2)C(C)(C)C)O)=C1O KGRVJHAUYBGFFP-UHFFFAOYSA-N 0.000 description 1
- RJASFPFZACBKBE-UHFFFAOYSA-N 2-Methylpropyl phenylacetate Chemical compound CC(C)COC(=O)CC1=CC=CC=C1 RJASFPFZACBKBE-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 235000009051 Ambrosia paniculata var. peruviana Nutrition 0.000 description 1
- 235000003097 Artemisia absinthium Nutrition 0.000 description 1
- 241000092668 Artemisia capillaris Species 0.000 description 1
- 235000008658 Artemisia capillaris Nutrition 0.000 description 1
- 235000017731 Artemisia dracunculus ssp. dracunculus Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- DULCUDSUACXJJC-UHFFFAOYSA-N Ethyl phenylacetate Chemical compound CCOC(=O)CC1=CC=CC=C1 DULCUDSUACXJJC-UHFFFAOYSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 244000038594 Phyllanthus urinaria Species 0.000 description 1
- 241000233614 Phytophthora Species 0.000 description 1
- 241000233622 Phytophthora infestans Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000000954 anitussive effect Effects 0.000 description 1
- 230000001088 anti-asthma Effects 0.000 description 1
- 239000000924 antiasthmatic agent Substances 0.000 description 1
- 229940124584 antitussives Drugs 0.000 description 1
- 239000001138 artemisia absinthium Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- MAQMEXSLUSZDQM-UHFFFAOYSA-N diethoxymethylbenzene Chemical compound CCOC(OCC)C1=CC=CC=C1 MAQMEXSLUSZDQM-UHFFFAOYSA-N 0.000 description 1
- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000012259 ether extract Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Classifications
-
- 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
- A01N27/00—Biocides, pest repellants or attractants, or plant growth regulators containing hydrocarbons
-
- 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
- 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
- A01N35/04—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 aldehyde or keto groups, or thio analogues thereof, directly attached to an aromatic ring system, e.g. acetophenone; Derivatives thereof, e.g. acetals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
- C07C45/78—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C7/00—Purification; Separation; Use of additives
- C07C7/005—Processes comprising at least two steps in series
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- 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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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Wood Science & Technology (AREA)
- Plant Pathology (AREA)
- Zoology (AREA)
- Pest Control & Pesticides (AREA)
- General Health & Medical Sciences (AREA)
- Dentistry (AREA)
- Health & Medical Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Water Supply & Treatment (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses an application of a artemisia rupestris extract in preparing a preparation for preventing and treating tobacco black shank, and belongs to the field of application of plant extracts. The main active components of the artemisia rupestris extract of the invention are as follows: the artemisia rupestris L extract is extracted from artemisia rupestris L by adopting an ethanol reflux extraction method; the artemisia rupestris extract is used for preventing and treating tobacco black shank and preparing corresponding medicines; experimental results show that the artemisia rupestris extract has a good inhibiting effect on tobacco black shank, can provide a new thought for preventing and treating the tobacco black shank, and has the characteristics of high efficiency, low toxicity and environmental friendliness.
Description
Technical Field
The invention relates to application of a artemisia rupestris extract in preparation of a preparation for preventing and treating tobacco black shank, and belongs to the field of application of plant extracts.
Background
The common sargassum herb is called tea velvet, and the whole herb is used as a medicine, and generally, the common sargassum herb is on the order of 1900-2300 m; distributed in northeast, north China, northwest and southwest provinces; has Wen Xin and has antitussive, expectorant, antiasthmatic, and heat and toxic materials clearing away effects; the researches on the artemisia rupestris are mainly focused on the field of medicine, relatively few researches on agriculture are carried out, and no report on the researches on the artemisia rupestris for inhibiting tobacco black shank is currently available.
The tobacco black shank causes great loss to the tobacco leaf production in each place every year, is one of important soil-borne rhizome diseases in the flue-cured tobacco production, and is very difficult to control in production; the active ingredients of the botanical pesticide are derived from the nature, have unique metabolic pathways, cannot cause environmental pollution, not only have the activities of sterilization and disinsection, but also can regulate plant growth and the like, can not kill natural enemies in the action process, are relatively safe to non-target organisms, and are not easy to generate drug resistance to harmful organisms, thereby solving a series of problems generated in the use process of chemical agents; the plant is a natural treasury of bioactive compounds, the active ingredient of the plant-derived pesticide is a natural active substance, and the plant-derived pesticide has the advantages of quick response, easy degradation, difficult generation of drug resistance of harmful organisms, safety to non-target organisms, environmental friendliness and the like, is paid attention to, and gradually forms a special chemical defense substance for resisting plant diseases and insect pests in long-term evolution and natural selection; therefore, there is an urgent need to find a plant source compound for preventing and controlling tobacco black shank.
Disclosure of Invention
In order to solve the technical problems, the invention aims at providing an application of a artemisia rupestris extract in preparation of a preparation for preventing and treating tobacco black shank, wherein the artemisia rupestris extract comprises the following main active components: the structural formula of the caryophyllene and/or the capillaris diacetylenone is as follows:
the structural formula of the capillaris diacetylenone is as follows:
the artemisia rupestris L extract is extracted from artemisia rupestris L by adopting an ethanol reflux extraction method, and the extraction method is a conventional method.
Preferably, the preparation method of the artemisia rupestris extract specifically comprises the following steps:
(1) Air drying aerial parts of Artemisia Bulleyana, pulverizing, and reflux extracting with ethanol.
(2) Mixing the extractive solutions, concentrating under reduced pressure to obtain extract, and collecting ethanol extract.
(3) The obtained ethanol extract of the artemisia rupestris is fully dissolved by distilled water, is extracted by petroleum ether, and is separated by silica gel column chromatography.
As a further preferred aspect of the present invention, the extraction in step (1) is carried out 3 to 4 times, each time for 3.5 to 4.5 hours.
As a preferred embodiment of the present invention, the artemisia rupestris extract is used for controlling tobacco black shank.
As a preferred embodiment of the invention, the artemisia rupestris extract is prepared into a drug for preventing and treating tobacco black shank by adopting a conventional method and adding conventional auxiliary materials, and the dosage form of the drug is not limited and can be liquid or powder.
As a preferred embodiment of the invention, the caryophyllene and the capillone (the mass ratio of the caryophyllene to the capillone is arbitrary) in the artemisia rupestris extract are used for preventing and treating tobacco black shank, and the caryophyllene and the capillone extracted from other plants or the artificially synthesized caryophyllene and capillone can also be used for preventing and treating tobacco black shank.
As a preferred embodiment of the invention, the preparation of the medicine for preventing and treating tobacco black shank by adding the conventional auxiliary materials into the artemisia rupestris extract with the conventional method, wherein the mass ratio of the artemisia rupestre to the artemisia rupestris extract is arbitrary, and the artemisia rupestre and the artemisia rupestris extract or the artificially synthesized artemisia rupestris and the artemisia rupestris extract can also be used for preparing the medicine for preventing and treating tobacco black shank.
As a preferred embodiment of the invention, the biggest herb in the artemisia rupestris extract is used for preventing and controlling tobacco black shank, and the biggest herb extracted from other plants or the biggest herb synthesized by other plants can also be used for preventing and controlling tobacco black shank.
As a preferred embodiment of the invention, the caryophyllus in the artemisia rupestris extract is prepared into a medicine for preventing and treating tobacco black shank by adopting a conventional method and adding conventional auxiliary materials, and the caryophyllus extracted from other plants or artificially synthesized caryophyllus can also be used for preparing the medicine for preventing and treating tobacco black shank.
As a preferred embodiment of the present invention, the capillarinone in the artemisia rupestris extract is used for controlling tobacco black shank, and capillarinone extracted from other plants or capillarinone artificially synthesized can also be used for controlling tobacco black shank.
As a preferred embodiment of the invention, the capillarinone in the artemisia rupestris extract is prepared into a medicine for preventing and treating tobacco black shank by adopting a conventional method and adding conventional auxiliary materials, and the capillarinone extracted from other plants or the capillarinone artificially synthesized can also be used for preparing the medicine for preventing and treating tobacco black shank.
The invention has the beneficial effects that:
the invention extracts and separates chemical components in the artemisia rupestris, and then adopts a toxic flat plate method and a growth rate method to screen the bacteriostatic activity of the potato late blight and the tobacco black shank, and the result shows that the artemisia rupestris extract has good inhibitory effect on the tobacco black shank, can provide a new thought for preventing and controlling the tobacco black shank, and has the characteristics of high efficiency, low toxicity and environmental friendliness.
Drawings
FIG. 1 is a mass spectrum of the Artemisia annua extract;
FIG. 2 is a graph showing the inhibitory effect of the Artemisia annua extract on tobacco black shank pathogenic bacteria;
FIG. 3 shows the rate of inhibition of Phytophthora nicotianae at a concentration of 0.2mg/mL of Artemisia niruri extract;
FIG. 4 shows the antibacterial rate of virgate wormwood herb diynone against Phytophthora nicotianae;
FIG. 5 shows the antibacterial rate against Phytophthora nicotianae at a concentration of cemetery of 10. Mu.g/ml;
FIG. 6 is a graph showing the effect of capillarity on the inhibition of Phytophthora nicotianae;
FIG. 7 is a graph showing the inhibitory effect of the cultivation of the moxarene at different concentrations for 4 days.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the scope of the present invention is not limited to the above.
Example 1
1. The preparation method of the artemisia rupestris extract specifically comprises the following steps of:
(1) Air drying and pulverizing aerial parts of radix et rhizoma Rhei Hao, extracting with ethanol under reflux for 3 times each for 4 hr.
(2) Mixing the 3 times of extractive solutions, concentrating under reduced pressure with rotary evaporator to obtain ethanol extract.
2. The determination of the active ingredients of the artemisia rupestris extract specifically comprises the following steps:
(1) The ethanol extract of artemisia rupestris obtained in this example was sufficiently dissolved in distilled water, extracted with petroleum ether, and the petroleum ether extract was separated by silica gel column chromatography.
(2) Gradient elution is adopted, 100% petroleum ether, petroleum ether with different proportions, ethyl acetate (100:1, 50:1, 25:1, 10:1, 5:1, 3:1, 2:1 and 1:1) and 100% ethyl acetate are sequentially used for elution, and the same components are combined through TLC analysis to obtain 18 components.
(3) And (3) screening the 18 components for antibacterial activity, and selecting the component with the best antibacterial effect.
(4) The GC-MS combined technology is adopted to analyze the chemical composition and the content of the components with the best antibacterial effect.
The specific experimental method comprises the following steps:
gas chromatography conditions: the carrier gas is helium, the flow rate of the column is 1.2mL/min, and the temperature of the sample inlet is: 250 ℃; the split ratio is 200:1, and the detector temperature is 220 ℃; the temperature-raising program is as follows: the initial temperature is 40 ℃, the temperature is kept for 1min, then the temperature is increased to 70 ℃ at 3 ℃/min, the temperature is kept for 1min, and then the temperature is increased to 220 ℃ at 5 ℃/min, and the temperature is kept for 10min; the sample loading was 1. Mu.L.
Mass spectrometry conditions: the interface temperature is 220 ℃; ion source temperature 200 ℃; an electron bombardment (EI) ion source; electron energy 70eV; the mass scanning range m/z is 5-450; comparing the mass spectrograms (shown in figure 1) of the components with reference substances in NIST107 and NIST21 mass spectrograms, taking similarity larger than 80%, and identifying the components by reference documents, wherein the relative percentage content of each component is determined by adopting a peak area normalization method.
2. Experimental results
TABLE 1 analysis of the composition of the components with the best antibacterial effect
| Sequence number | Peak time/min | Names of Compounds | Molecular formula | Relative content% |
| 1 | 10.66 | dihydrotagetone | C 10 H 18 O | 0.35 |
| 2 | 16.73 | Benzaldehyde diethyl acetal | C 11 H 16 O 2 | 0.20 |
| 3 | 17.31 | Phenylacetic acid ethyl ester | C 10 H 12 O 2 | 0.71 |
| 4 | 23.41 | Phenylacetic acid isobutyl ester | C 12 H 16 O 2 | 0.26 |
| 5 | 25.39 | Eben-zene | C 12 H 10 | 49.11 |
| 6 | 29.55 | Capillary artemisia diacetylenone | C 12 H 8 O | 33.63 |
| 7 | 52.25 | 2,2' -methylenebis- (4-methyl-6-tert-butylphenol) | C 23 H 32 O 2 | 2.75 |
From table 1, it can be found that the compounds with relatively high content in the active ingredient are: the moxarene and the capillaris diynone; the extract has the main components of the caryophyllus communis which are taken as active ingredients, wherein the relative content of the caryophyllus communis is 49.11 percent, the relative content of the capillaris diynone is 33.63 percent, and the active ingredients of the caryophyllus communis and the capillaris diynone are found to be effective when the extract is used singly or mixed in different proportions.
3. The prevention and treatment effect of the artemisia rupestris extract on the tobacco black shank is verified, and the phytophthora nicotianae is a main cause of the caused tobacco black shank, so that the prevention and treatment effect of the artemisia rupestris extract on the tobacco black shank is further proved by researching the inhibition effect of the artemisia rupestris extract on the phytophthora nicotianae, and the specific steps are as follows:
(1) The artemisia rupestris extract is prepared into a mother solution of 100 mug/mL by using dimethyl sulfoxide (DMSO) and the mother solution is sterilized by a pasteurization method for standby.
(2) A rye culture medium was prepared at a ratio of 60g of rye, 18g of agar, 15g of white sugar and 1000mL of water.
(3) And autoclaving the prepared culture medium and glassware for standby.
(4) Adding a certain amount of mother liquor into a rye culture medium to be cooled under aseptic condition, fully mixing to make the liquid medicine containing different concentrations of each component in the culture medium, taking the rye culture medium containing equivalent DMSO solvent as a blank Control (CK), inoculating 1 bacterial cake (phi=5 mm) in the center of each dish after the culture medium is solidified, and repeating each treatment for 3 times; culturing in an electrothermal constant temperature incubator at 18deg.C, measuring the diameter of each treated colony by crisscross method every day from the 2 nd day after inoculation, calculating antibacterial rate until the control group grows up to the plate, and stopping counting.
Pure growth = colony mean diameter-5
The control pure growth amount of the invention is as follows: subtracting the diameter of the bacterial cake from the average diameter of the colony of the control group; the pure growth amount of the treatment is as follows: the average diameter of the treated colonies minus the diameter of the patties.
(5) Toxicity measurement: converting the calculated bacteriostasis rate of each concentration into a mechanical value, taking the logarithm of the concentration as an X axis, taking the mechanical value of the bacteriostasis rate as a Y axis, and drawing by Excel to obtain a virulence equation to obtain EC 50 。
TABLE 2 antibacterial Rate of Artemisia rupestris extract against Phytophthora nicotianae
Table 3 determination of virulence of artemisia rupestris extract on phytophthora nicotianae
Analysis of experimental results:
fig. 2 is a graph showing the inhibitory effects of the artemisia rupestris extract on pathogenic bacteria of tobacco black shank, wherein the graphs (a), (b), (c), (d) and (e) respectively show that the concentration of the artemisia rupestris extract is 0, 20, 40, 80 and 160 mug/mL, and the inhibitory effects of the artemisia rupestris extract on the flat plate of a control group (graph a) are shown by the graph, and the higher the concentration, the more remarkable the inhibitory effects of the artemisia rupestris active component on the phytophthora rupestis.
From Table 2, it can be seen that the active components of the artemisia rupestris have certain inhibition effect on the phytophthora nicotianae at the concentrations of 0, 20, 40, 80 and 160 mug/mL, and the higher the concentration is, the more obvious the inhibition effect is, and the obvious dosage effect exists; the antibacterial rate of the artemisia rupestris extract at the same concentration is slightly reduced along with the extension of the culture time, and the antibacterial rate of a treatment group with 160 mug/mL is still more than 50% when the 6d control is cultivated to grow up to a flat plate; therefore, the artemisia rupestris extract has strong antibacterial activity on phytophthora nicotianae.
From Table 3, it can be seen that the effective medium concentration of the artemisia rupestris extract (EC 50 ) Between 18.085 μg/mL and 132.148 μg/mL, with the virulence determined by culture 2d being strongest; when the phytophthora nicotianae silk on the control flat plate is full, the hypha growth of the treatment group is obviously inhibited, and the higher the concentration is, the better the inhibition effect on the phytophthora nicotianae silk growth is; microscopic observation shows that the mycelium morphology of the active component treatment group is obviously changed, rupture and distortion occur, the integrity of cell membranes is damaged, and the spore yield is obviously reduced compared with that of a control group, so that the active component achieves the antibacterial effect by inhibiting the normal growth of mycelium, destroying the thallus structure, inhibiting the spore production of bacteria and the like.
Fig. 3 shows the antibacterial rate of the increased dose, namely the antibacterial rate of the artemisia rupestris when the concentration of the artemisia rupestris extract is 0.2mg/mL, and as can be seen from the graph, the phytophthora nicotianae hyphae grows on the control plate in 2 days, the hyphae growth of the treatment group is inhibited, and no hyphae are found, so that the antibacterial efficiency is 100%, the hyphae of the treatment group slowly grow out along with the increase of time, but compared with the control group, the growth of the hyphae is obviously inhibited, the antibacterial effect still exists on the 6 th day, and the antibacterial efficiency is weakened along with the increase of time.
The experiment shows that the artemisia rupestris extract has very good placing effect on phytophthora nicotianae, and the phenomena of mycelium rupture, mycelium distortion, cell membrane integrity damage, spore yield reduction and the like are found after the extract is treated; the extract can be used for inhibiting the normal growth of phytophthora nicotianae, destroying the thallus structure, inhibiting the spore production of pathogenic bacteria and the like.
Example 2
In order to further prove the active ingredients of the artemisia rupestris extract, the invention adopts a growth rate method to measure the antibacterial toxicity of the artemisia rupestris to the phytophthora nicotianae through artificially synthesizing the caryophyllene and the capillaris diacetylenone.
In the embodiment, the bacteriostasis experiment is carried out on phytophthora nicotianae by adopting artificially synthesized caryophyllene and capillaris diynone, the experimental method is the same as that in the embodiment 1, and the results are shown in fig. 4 and 5.
Fig. 4 shows the antibacterial rate of phytophthora nicotianae at a concentration of 10 μg/ml of capillaris, and shows that the phytophthora nicotianae hyphae grow on the control plate at 2 days due to the relatively short time, the hypha growth of the treatment group is inhibited and does not grow, so that the antibacterial efficiency is 100%, the hypha of the treatment group grows slowly with the increase of time, compared with the control group, the growth of the hypha is obviously inhibited, the antibacterial effect still exists at 6 days, and the antibacterial efficiency is weakened with the increase of time.
Fig. 5 shows the antibacterial rate of Phytophthora nicotianae at a concentration of 10 μg/ml, and it can be seen from the graph that the Phytophthora nicotianae has a certain inhibition effect on Phytophthora nicotianae, and the antibacterial efficiency is reduced with the increase of time.
The inhibition effect graphs of the capillarity ketone concentrations of 0, 5, 10, 15 and 20 mug/mL from left to right in FIG. 6 are respectively shown, and the inhibition effect graph shows that the capillarity ketone has a certain inhibition effect on two strains of the phytophthora nicotianae when the concentration is 5, 10, 15 and 20 mug/mL on the (CK) flat plate of the control group and the inhibition effect is more obvious when the concentration is higher.
From left to right in FIG. 7, the inhibition effect graphs of the concentration of the moxarene at 0, 10, 25, 40, 55 and 70 mug/mL are shown, and the inhibition effect graph shows that the moxarene has a certain inhibition effect on two strains of phytophthora nicotianae when the concentration of the moxarene at 10, 25, 40, 55 and 70 mug/mL on the (CK) flat plate of the control group is higher, and the inhibition effect is more obvious when the concentration is higher.
The results show that both the pure caryophyllene and the pure capillaris diynone have an inhibiting effect on phytophthora nicotianae.
The main active components of the extract are the caryophyllene and the capillaris diacetylenone, and the caryophyllene and the capillaris diacetylenone are used independently or mixed in different proportions, so that the extract has remarkable antibacterial effect, and the phenomena of mycelium rupture, mycelium distortion, cell membrane integrity damage, spore yield reduction and the like can be found after the treatment of the extract by the medicament; the compound can be used for inhibiting the normal growth of phytophthora nicotianae, destroying the thallus structure, inhibiting the spore production of pathogenic bacteria and the like.
Claims (4)
1. The application of the artemisia rupestris extract in preparing a preparation for preventing and treating tobacco black shank comprises the following main active components: the structural formulas of the caryophyllene and the capillaris diacetylenone are as follows:the method comprises the steps of carrying out a first treatment on the surface of the The structural formula of the capillaris diacetylenone is as follows: />The preparation method of the artemisia rupestris extract specifically comprises the following steps:
(1) Air drying aerial parts of Artemisia Bulleyana, pulverizing, and reflux extracting with ethanol;
(2) Mixing the extractive solutions, concentrating under reduced pressure with rotary evaporator to obtain ethanol extract.
2. The use of the artemisia rupestris extract according to claim 1 for preparing a preparation for preventing and treating tobacco black shank, wherein the artemisia rupestris extract is extracted 3-4 times in step (1) for 3.5-4.5 hours each time.
3. Use of an artemisia rupestris extract according to claim 1 for the preparation of a formulation for controlling tobacco black shank, characterized in that: the artemisia rupestris extract is directly used for preventing and treating tobacco black shank.
4. Use of an artemisia rupestris extract according to claim 3 for the preparation of a formulation for controlling tobacco black shank, characterized in that: the artemisia rupestris extract is prepared into a medicine for preventing and treating tobacco black shank by adopting a conventional method and adding conventional auxiliary materials.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100102411A (en) * | 2009-03-11 | 2010-09-24 | 영농조합법인 초락도 약쑥작목반 | Agricultural composition comprising wormwood extracts |
| CN104938525A (en) * | 2015-05-26 | 2015-09-30 | 湖北省烟草科学研究院 | Preparation method and application of plant fungicide |
| WO2019180420A1 (en) * | 2018-03-19 | 2019-09-26 | M H Poskitt Limited | Pesticide |
-
2023
- 2023-09-20 CN CN202311217689.9A patent/CN116965407B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100102411A (en) * | 2009-03-11 | 2010-09-24 | 영농조합법인 초락도 약쑥작목반 | Agricultural composition comprising wormwood extracts |
| CN104938525A (en) * | 2015-05-26 | 2015-09-30 | 湖北省烟草科学研究院 | Preparation method and application of plant fungicide |
| WO2019180420A1 (en) * | 2018-03-19 | 2019-09-26 | M H Poskitt Limited | Pesticide |
Non-Patent Citations (8)
| Title |
|---|
| 27种植物提取物抗烟草花叶病毒活性分析;左安建;宗同铠;谭亚婷;何彦仪;李雅丽;闫俊伊;李晋芳;闫晓慧;;西南林业大学学报(自然科学)(第05期);99-105 * |
| 5种蒿属植物提取物对马铃薯晚疫病的抑菌活性研究;何彦仪等;西南林业大学学报(自然科学);162-167 * |
| Pesticidal activities and active ingredients of Artemisia;Zhou LiJuan等;Acta Agriculturae Universitatis Jiangxiensis;699-705 * |
| 烟草黑胫病绿色防控技术研究进展;王正浩;王维超;毋彦彬;姚云鹏;张朝阳;蔡宝才;孟黎明;;安徽农业科学(第10期);26-29 * |
| 烟草黑胫病防治研究进展;高文华;李江美;;热带农业科技(第01期);54-58 * |
| 臭蒿水乳剂抗烟草黑胫病的作用;徐清辉;胡国元;杨春雷;余君;杨锦鹏;;湖北大学学报(自然科学版)(第01期);55-60 * |
| 茵陈蒿的化学成分和生物学活性研究概述;王兰, 陈炅然, 陶菊蓉;中兽医医药杂志(第01期);43-44 * |
| 蒿属植物的农药活性及其有效成分;周利娟;桑晓清;孙永艳;杨文杰;;江西农业大学学报(第04期);86-92 * |
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