CN102283256A - Star anise extract for killing Meloidogyne spp. and preparation method thereof - Google Patents
Star anise extract for killing Meloidogyne spp. and preparation method thereof Download PDFInfo
- Publication number
- CN102283256A CN102283256A CN2011102140370A CN201110214037A CN102283256A CN 102283256 A CN102283256 A CN 102283256A CN 2011102140370 A CN2011102140370 A CN 2011102140370A CN 201110214037 A CN201110214037 A CN 201110214037A CN 102283256 A CN102283256 A CN 102283256A
- Authority
- CN
- China
- Prior art keywords
- ethyl alcohol
- chinese anise
- absolute ethyl
- extract
- nematode
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a star anise extract for killing Meloidogyne spp. and a preparation method thereof, and provides a method for preparing a star anise absolute ethanol extract. The method comprises the following steps of: 1) uniformly mixing star anise and absolute ethanol to obtain a mixture; and 2) performing ultrasonic treatment on the mixture obtained in step 1), and suction-filtering and concentrating to obtain the star anise absolute ethanol extract. Through the experiment, star anise is extracted by using absolute ethanol, the poisoning effect of extracting solution on the Meloidogyne spp. is tested, and the result is that the extract has strong poisoning activity on the Meloidogyne spp.; therefore, the extract lays a foundation for the development and application of Meloidogyne spp. biocontrol preparations in the future. In the method, an active substance for killing the Meloidogyne spp. is extracted by using a natural harmless plant material, so that the pollution caused by chemical pesticides can be reduced; and the extraction method is simple, and can be easily popularized and applied to the control of plant rootknot nematodes.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of Chinese anise nematicide extract and preparation method thereof.
Background technology
Root-knot nematode (Meloidogyne spp.) is the root system of plant obligate parasite, is the current important pathogen of a class that influences agricultural production.
Since the eighties in 20th century, utilize the research of plant extraction liquid control nematode to cause people's attention gradually, so far the nematicide plant of having reported has 102 sections 226 to belong to 316 kinds approximately, wherein study more have composite family, Meliaceae, pulse family, Cruciferae, Solanaceae, grass family, Euphorbiaceae, Papaveraceae, Apocynaceae, Mimosaceae, Liliaceae etc., but the natural plant resource is very abundant, many plants with nematocidal effect are also undiscovered, " Chinese poisonous plant " book is listed poisonous plant kind more than 1300 in, wherein, numerous species all has insecticidal activity.And a lot of plants can be gathered materials on the spot, it is convenient to use, and effect is obvious, therefore, there is more research at present domestic screening and aspect such as cytotoxicity mensuration to the nematicide plant resources, fully investigates and excavate the direction that how more effective new nematicide floristics will become future development.
Chinese anise is the fruit of Magnoliaceae Chinese anise plant (Illicium verum).Gather autumn, dries to be used as medicine." national Chinese herbal medicine compilation " described its fruit and contained volatile oil 4～9%, generally is about 5%, fat oil about 22% and protein, natural gum and resin etc.Main Ingredients and Appearance is that anisole (anethole) is about 80～90% in the volatile oil, and all the other are α-terpineol, d-firpene and a small amount of estragole (methyl chavicol) etc.Its active component is used more on physianthropy, and the ethanol extract of Chinese anise all has stronger inhibitory action to staphylococcus aureus, comma bacillus, typhoid bacillus, pneumococcus, diphtheria bacillus, shigella dysenteriae, Escherichia coli and some common pathogenic dermatophytes.In plant application medically, have only a small amount of research report of control pest of stored grain and pest diamond-back moth, in the sick control of plant root-knot nematode, do not appear in the newspapers.
Summary of the invention
An object of the present invention is to provide a kind of method for preparing Chinese anise absolute ethyl alcohol extract.
Method provided by the invention comprises the steps:
1) with Chinese anise and absolute ethyl alcohol mixing, obtains mixture;
2) mixture that step 1) is obtained carries out ultrasonic processing, promptly obtains Chinese anise absolute ethyl alcohol extract.
Described Chinese anise is the fruit of Chinese anise.
In the step 1), the proportioning of described Chinese anise and absolute ethyl alcohol is 10g: (45-55) ml;
Step 2) in, described ultrasonic power is 300-500W, and described ultrasonic operating frequency is 28KHz-100KHz, and the described ultrasonic time is 25min-35min; Described ultrasonic temperature is 23 ℃-28 ℃.
In the step 1), the proportioning of described Chinese anise and absolute ethyl alcohol is 10g: (45,50 or 55) ml;
Step 2) in, described ultrasonic power is 300W, 400W or 500W, and described ultrasonic operating frequency is 28KHz, 45KHz or 100KHz, and the described ultrasonic time is 25min, 30min or 35min; Described ultrasonic temperature is 23 ℃, 25 ℃ or 28 ℃.
Before step 1), also comprise the step that described Chinese anise is crushed to 35 orders-45 order size; Described Chinese anise is crushed to and is specially 35 orders, 40 orders or 45 order sizes;
In step 2) after, also comprise the steps: the ultrasonic product that described ultrasonic processing obtains is concentrated into paste through suction filtration, filtrate successively, obtain Chinese anise absolute ethyl alcohol extract.
Chinese anise absolute ethyl alcohol extract by described method preparation also is a scope of protection of the invention.
The application of described Chinese anise absolute ethyl alcohol extract in nematicide and/or anti-nematode also is the scope of protection of the invention.
The application of described Chinese anise absolute ethyl alcohol extract in the biological prevention and control agent of preparation nematode also is the scope of protection of the invention.
Described nematode is root-knot nematode (Meloidogyne spp.).
Another object of the present invention provides the biological prevention and control agent of a kind of nematode.
Biological prevention and control agent provided by the invention, its active component are described Chinese anise absolute ethyl alcohol extract.
Described nematode is root-knot nematode (Meloidogyne spp.).
The present invention of experiment showed, of the present invention adopts absolute ethyl alcohol that Chinese anise is extracted, and with the toxic effect test of extract to root-knot nematode, the result has strong cytotoxicity for this extract to root-knot nematode; Illustrate that extract of the present invention lays the foundation for the development and application of root-knot nematode biological prevention and control agent from now on.
Embodiment
Employed experimental technique is conventional method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The acquisition of embodiment 1, Chinese anise absolute ethyl alcohol extract
High speed disintegrator, 6202 types, Beijing is encircled inferior Tianyuan Mechanical Technology Inc. and is made; Desk-top three frequency controlled ultrasonic wave washers, KQ-500VDV type, Kunshan Ultrasonic Instruments Co., Ltd.; Rotary Evaporators, RE-2000 type, Shanghai Yarong Biochemical Instrument Plant; Stereomicroscope, Motic SMZ-140 type, Maike Aodi Industry Group Co Ltd; The trace adjustable pipette, GILSON, France; The disposable Tissue Culture Plate in 96 holes, JET BIOFIL
Type, Canada; Electro-heating standing-temperature cultivator, SKP-02.420 type, Hengfeng Medical Instruments Co., Ltd., Huangsih City; Absolute ethyl alcohol (analyzing pure), the Beijing Chemical Plant;
Available from the big pharmacy of Beijing Tongrentang, belong to Magnoliaceae for the Chinese anise of trying (Illicium verum), use fruit.
Gather the old complaint of tomato root-knot eelworm disease from morbidity plot, field for examination root-knot nematode (Meloidogyne spp.), (two kinds of agricultural chemicals are to the Toxicity Determination of Meloidogyne incognita according to people such as horse Zhe, Beijing Agricultural College's journal, 2009,24 (3): the 17-19) method of Jie Shaoing, by choose egg capsule and hatching acquisition from old complaint.
1, the preparation of root-knot nematode second instar larvae suspension
Gather the old complaint of tomato root-knot eelworm disease from the field, get the tomato root system that produces a large amount of root knots, running water is rinsed well gently, with the dissecting needle milky egg capsule on the picking old complaint gently, put into the little culture dish of diameter 6cm, add a small amount of sterile water, in 25 ℃ of insulating boxs, hatch 3d, collect 2 instar larvaes (be called for short J2), and add a certain amount of sterile water and it is mixed with finite concentration (root-knot nematode second instar larvae suspension 500/mL) is standby.
2, the preparation of plant extracts
Method one:
1) pulverizes
The Chinese anise of drying is put into high speed disintegrator pulverizing back cross 40 mesh sieves, it is standby to put into sealed bag.
2) extract
Adopt ultrasonic extraction, accurately take by weighing the plant drymeal 10g that crushes, add absolute ethyl alcohol 50mL in iodine flask, put into supersonic generator, ultrasonic power is 400W, operating frequency is 45KHz, and ultrasonic temperature is 25 ℃, behind the ultrasonic processing 30min, obtain the absolute ethyl alcohol extract of Chinese anise, through suction filtration (use bottle,suction and Buchner funnel, fill up 2 metafiltration paper), then filtrate is evaporated to thick paste shape (Chinese anise absolute ethyl alcohol extract) in rotary evaporator.Before carrying out insecticidal activity assay, paste is added water be mixed with the aqueous solution that concentration is 20mg/mL, the brown wide-mouth bottle sealing of the ground of packing into then, it is standby to place 4 ℃ of refrigerators to preserve, and is plant extraction liquid (Chinese anise absolute ethyl alcohol extract).
Method two:
1) pulverize: basic identical with method one, different is to pulverize the back to cross 35 mesh sieves, and it is standby to put into sealed bag.
2) extract: basic identical with method one, that different is plant drymeal 10g, adds absolute ethyl alcohol 45mL, and ultrasonic power is 300W, and operating frequency is 28KHz, and ultrasonic temperature is 23 ℃, ultrasonic processing 25min; Obtain plant extraction liquid (Chinese anise absolute ethyl alcohol extract).
Method three:
1) pulverize: basic identical with method one, different is to pulverize the back to cross 45 mesh sieves, and it is standby to put into sealed bag.
2) extract: basic identical with method one, that different is plant drymeal 10g, adds absolute ethyl alcohol 55mL, and ultrasonic power is 500W, and operating frequency is 100KHz, and ultrasonic temperature is 28 ℃, ultrasonic processing 35min; Obtain plant extraction liquid (Chinese anise absolute ethyl alcohol extract).
Embodiment 2, cytotoxicity assay method
The employing method of tagging is measured, concrete steps are: get 96 porocyte culture plates, it is the plant extraction liquid 150 μ L that the method one by embodiment 1 step 2 of 20mg/mL obtains that every hole adds the concentration for preparing respectively, add isopyknic root-knot nematode second instar larvae suspension that obtains by step 1 again, mixing, 25 ℃ leave standstill, respectively under stereomicroscope, observing behind 12h, 24h and the 48h, check nematode (needle contact method) anyway, stiff or curling motionless polypide is considered as death, is calculated as follows the nematode corrected mortality.With the sterile water is contrast, and every processing repeats 3 times, results averaged ± standard error.
Data are handled and analyzed: experimental data is by the DPS2000 analysis and arrangement, the result by the multiple comparison method of the new multipole difference of Deng Kenshi (Duncan ' s multiple range test, DMRT) carry out significance of difference analysis.The strong and weak grade scale of nematicide is pressed (Chandravadana M V such as Chandravadana, Eugene S, Nidiry J, et al.Nematicidal activity of some plant extracts[J] .Indian J Nematol, 1996,26 (2): method 148-151.) is carried out, and concrete grade scale is: "-" expression non-activity, corrected mortality≤10%; "+" expression is weak active, and corrected mortality is 10%～30%; " ++ " expression medium activity, corrected mortality is 30%～50%; " +++" the strong activity of expression, corrected mortality is 50%～80%; " ++ ++ " the strong activity of expression, corrected mortality＞80%.Indicate different lowercase alphabet differentials different significantly (p＜0.05, DMRT method) after the same column data, capitalization is represented difference extremely significantly (p＜0.01, DMRT method).
When being 10mg/mL processing nematode with the extract concentrations, Chinese anise absolute ethyl alcohol extract is as follows to the cytotoxicity result of root-knot nematode:
After handling back 12h, Chinese anise absolute ethyl alcohol extract to the cytotoxicity intensity of root-knot nematode is ++ ++, corrected mortality is 97.7 ± 2.30aA,
After handling back 24h, Chinese anise absolute ethyl alcohol extract to the cytotoxicity intensity of root-knot nematode is ++ ++, corrected mortality is 98.85 ± 1.15aA,
After handling back 48h, Chinese anise absolute ethyl alcohol extract to the cytotoxicity intensity of root-knot nematode is ++ ++, corrected mortality is 100 ± 0.00aA.
From above result as seen, the absolute ethyl alcohol extract of Chinese anise shows strong cytotoxicity to root-knot nematode, and behind the processing 24h, the nematode corrected mortality is respectively 98.85%; After handling 48h, the nematode corrected mortality is respectively 100%.
Show also that according to the DPS The result of multiple comparisons absolute ethyl alcohol extract of Chinese anise difference on 0.05 and 0.01 level is not remarkable, all shows strong cytotoxicity.
Employing detects by the method two of embodiment 1 step 2 and the plant extraction liquid of method three acquisitions, result and method one no significant difference with quadrat method.
Claims (10)
1. a method for preparing Chinese anise absolute ethyl alcohol extract comprises the steps:
1) with Chinese anise and absolute ethyl alcohol mixing, obtains mixture;
2) mixture that step 1) is obtained carries out ultrasonic processing, promptly obtains Chinese anise absolute ethyl alcohol extract.
2. method according to claim 1 is characterized in that:
In the step 1), the proportioning of described Chinese anise and described absolute ethyl alcohol is 10g: (45-55) ml;
Step 2) in, described ultrasonic power is 300W-500W, and described ultrasonic operating frequency is 28KHz-100KHz, and the described ultrasonic time is 25min-35min; Described ultrasonic temperature is 23 ℃-28 ℃.
3. method according to claim 1 and 2 is characterized in that:
In the step 1), the proportioning of described Chinese anise and absolute ethyl alcohol is 10g: (45,50 or 55) ml;
Step 2) in, described ultrasonic power is 300W, 400W or 500W, and described ultrasonic operating frequency is 28KHz, 45KHz or 100KHz, and the described ultrasonic time is 25min, 30min or 35min; Described ultrasonic temperature is 23 ℃, 25 ℃ or 28 ℃.
4. according to arbitrary described method among the claim 1-3, it is characterized in that:
Before step 1), also comprise the step that described Chinese anise is crushed to 35 orders-45 order size; Described Chinese anise is crushed to and is specially 35 orders, 40 orders or 45 order sizes;
In step 2) after, also comprise the steps: the ultrasonic product that described ultrasonic processing obtains is concentrated into paste through suction filtration, filtrate successively, obtain Chinese anise absolute ethyl alcohol extract.
5. by the Chinese anise absolute ethyl alcohol extract of arbitrary described method preparation among the claim 1-4.
6. the application of the described Chinese anise absolute ethyl alcohol of claim 5 extract in nematicide and/or anti-nematode.
7. the application of the described Chinese anise absolute ethyl alcohol of claim 5 extract in the biological prevention and control agent of preparation nematode.
8. according to claim 6 or 7 described application, it is characterized in that: described nematode is root-knot nematode (Meloidogyne spp.).
9. the biological prevention and control agent of a nematode, its active component is the described Chinese anise absolute ethyl alcohol of claim 5 extract.
10. biological prevention and control agent according to claim 9 is characterized in that: described nematode is root-knot nematode (Meloidogyne spp.).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110214037 CN102283256B (en) | 2011-07-28 | 2011-07-28 | Star anise extract for killing Meloidogyne spp. and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110214037 CN102283256B (en) | 2011-07-28 | 2011-07-28 | Star anise extract for killing Meloidogyne spp. and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102283256A true CN102283256A (en) | 2011-12-21 |
| CN102283256B CN102283256B (en) | 2013-05-01 |
Family
ID=45330236
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110214037 Expired - Fee Related CN102283256B (en) | 2011-07-28 | 2011-07-28 | Star anise extract for killing Meloidogyne spp. and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102283256B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102894015A (en) * | 2012-11-01 | 2013-01-30 | 彭万喜 | Method for preparing antibacterial agent |
| CN109997882A (en) * | 2019-01-22 | 2019-07-12 | 郑州昊天化工产品有限公司 | The method that Chinese herbal medicine prevents and treats root nematode by spraying |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102018266A (en) * | 2009-09-11 | 2011-04-20 | 天津科技大学 | All-natural plant composite preservative and preparation method and preservation method thereof |
| CN102031196A (en) * | 2010-12-02 | 2011-04-27 | 北京航空航天大学 | Method for extracting effective ingredients in industrial residue of Chinese anise |
-
2011
- 2011-07-28 CN CN 201110214037 patent/CN102283256B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102018266A (en) * | 2009-09-11 | 2011-04-20 | 天津科技大学 | All-natural plant composite preservative and preparation method and preservation method thereof |
| CN102031196A (en) * | 2010-12-02 | 2011-04-27 | 北京航空航天大学 | Method for extracting effective ingredients in industrial residue of Chinese anise |
Non-Patent Citations (1)
| Title |
|---|
| 徐芃等: "索式提取与超声波提取对茴香抑菌活性成分的影响", 《中国医药导报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102894015A (en) * | 2012-11-01 | 2013-01-30 | 彭万喜 | Method for preparing antibacterial agent |
| CN109997882A (en) * | 2019-01-22 | 2019-07-12 | 郑州昊天化工产品有限公司 | The method that Chinese herbal medicine prevents and treats root nematode by spraying |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102283256B (en) | 2013-05-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lazreg Aref et al. | In vitro cytotoxic and antiviral activities of Ficus carica latex extracts | |
| Pinto et al. | Identification and structural characterization of novel cyclotide with activity against an insect pest of sugar cane | |
| Argentieri et al. | Evaluation of nematicidal properties of saponins from Medicago spp. | |
| Bajwa | Appraisal of antifungal activity of Aloe vera | |
| Mavundza et al. | Larvicidal activity against Anopheles arabiensis of 10 South African plants that are traditionally used as mosquito repellents | |
| Shukla et al. | Hepatoprotective and antioxidant activities of crude fractions of endophytic fungi of Ocimum sanctum Linn. in rats | |
| CN103636660A (en) | Environmental-friendly particulate agent for controlling sugarcane pests | |
| CN102246831B (en) | Peony bark nematicidal extract and preparation method thereof | |
| Santana et al. | Antitermitic activity of extractives from three Brazilian hardwoods against Nasutitermes corniger | |
| CN103609597A (en) | Botanical synergistic compound pesticide | |
| Devakumar et al. | Nematicidal principles frcm neem (Azadirachta indica A. Juss). Part I. Screening of neem kernel fractions against Meloidogyne incognita | |
| Cheng et al. | Optimization of the microemulsion formulation of curcuma oil and evaluation of its acaricidal efficacy against Tetranychus cinnabarinus (Boisduval)(Acari: Tetranychidae) | |
| Caputo et al. | Mentha pulegium L.: A plant underestimated for its toxicity to be recovered from the perspective of the circular economy | |
| CN102283256B (en) | Star anise extract for killing Meloidogyne spp. and preparation method thereof | |
| Younus et al. | Extracts of Euphorbia nivulia Buch.-Ham. showed both phytotoxic and insecticidal capacities against Lemna minor L. and Oxycarenus hyalinipennis Costa | |
| Francine et al. | Larvicidal activities of hydro-ethanolic extracts of three Cameroonian medicinal plants against Aedes albopictus | |
| Lim et al. | Mosquito larvicidal activity and cytotoxicity of the extracts of aromatic plants from Malaysia | |
| CN102246829B (en) | Clove extract for killing nematode and preparation method thereof | |
| CN101336644A (en) | Plant insecticide prepared by humulus and its preparing process horehound | |
| Caramelo et al. | Potential applications of the Cytisus shrub species: Cytisus multiflorus, Cytisus scoparius, and Cytisus striatus | |
| Cornara et al. | Micromorphological investigation on the leaves of the rock samphire (Crithmum maritimum L.): Occurrence of hesperidin and diosmin crystals | |
| CN102318651B (en) | Nematicidal extract of radix-polygoni multiflori and preparation method thereof | |
| CN101352179A (en) | Potentiating agent of plant source pesticides | |
| Teklani et al. | Mosquito repellent and larvicidal activities of Acalypha indica leaf extracts | |
| CN107897217A (en) | A kind of natural volatile mosquito driving and killing agent of plant source property and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20111221 Assignee: The middle peasants lvkang Biotechnology Co. Ltd. (Beijing) Assignor: Beijing University Of Agriculture Contract record no.: 2013990000869 Denomination of invention: Star anise extract for killing Meloidogyne spp. and preparation method thereof Granted publication date: 20130501 License type: Exclusive License Record date: 20131224 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130501 Termination date: 20150728 |
|
| EXPY | Termination of patent right or utility model |