CN101385471A - Active material extracted from Acutangular Anisodus root and uses thereof - Google Patents
Active material extracted from Acutangular Anisodus root and uses thereof Download PDFInfo
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- CN101385471A CN101385471A CNA2008100589788A CN200810058978A CN101385471A CN 101385471 A CN101385471 A CN 101385471A CN A2008100589788 A CNA2008100589788 A CN A2008100589788A CN 200810058978 A CN200810058978 A CN 200810058978A CN 101385471 A CN101385471 A CN 101385471A
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Abstract
The invention relates to an active matter which is extracted from acutangular anisodus root and the application thereof, which belongs to the technical field of biological pesticide. The active matter is obtained by the following steps: a. the collected entire plant of acutangular anisodus root is washed by water, and then the washed acutangular anisodus root is put into an oven and baked for 10 hours at the temperature of 50 DEG C; b. the baked acutangular anisodus root is sheared into small sections with the length of 0.5cm; c.10g of the sheared acutangular anisodus root is weighed is taken and placed into 150ml of triangular flasks respectively; wherein, one flask is added with 100ml of 100 percent acetone for extracting, and the other flask is added with 100ml of 100 percent methanol for extracting; the samples extracted by the two solvents are placed for 72 hours at the room temperature; d. the leach liquor of the samples processed under different conditions is respectively filtered, and concentrated at the temperature of 50 DEG C so as to obtain active extract of the acutangular anisodus root; and the extract of acetone is dissolved by the acetone, the extract of methanol uses the methanol to dissolve the sample, the quantity is fixed to be 0.1g/ml and the obtained sample is put into the refrigerator for spare. The active extract of the invention has the advantages of simple and convenient preparation, high efficiency and low toxicity and the like, and can be applied to preparing biological pesticide used for killing nematodes.
Description
Technical field:
The present invention relates to a kind of active matter that from scopolia acutangula, extracts and application, belong to biological pesticide technical field.
Background technology:
Pine wood nematode (Bursaphelenchus xylophilus) belongs to Nematoda (Nematoda), Tylenchida (Tylenchida), Aphelenchidae (Aphelenchoididae), Bursaphelenchus (Bursaphelenchus).The pine nematode that is caused by pine wood nematode is one of forest disease of tool danger in the world, is a kind of destructive popular disease of pine tree, can cause pine forest withered in flakes, is called as " cancer " of pine tree.This disease period of disease weak point, dead fast, the easy propagation of disease tree, difficult control.Therefore, pine wood nematode (B.xylophilus) agent of killing of research and development high-efficiency low-toxicity is subjected to common attention.Plant is one of main source that produces bioactive natural product, and seeking to have from plant has the natural metabolite of cytotoxicity to become one of emphasis of biocontrol of nematodes research to pine wood nematode (B.xylophilus).
Scopolia acutangula (Anisodus acutangulus
.Y.Wu et C.Chen), mainly be distributed in the Yunnan Province of China northwestward for the Solanaceae Tangut Anisodus Radix belongs to (Anisodus) herbaceos perennial.The effects of have the analgesia, control traumatic injury, dispelling rheumatism etc. are spasmolysis and analgesia medicine commonly used among the people.But this plant useless so far has active report to nematode.
Summary of the invention:
The objective of the invention is from scopolia acutangula (A.acutangulus), to extract activity extract, be used for preparation and develop the biologic product that high-efficiency low-toxicity kills pine wood nematode (B.xylophilus) with strong inhibition pine wood nematode (B.xylophilus) effect.
The present invention is achieved in that
1, preparation activity extract
(1) sample collecting and preliminary treatment: the scopolia acutangula complete stool that collects is put into baking oven after with the running water flushing, and baking is 10 hours under 50 ℃ of temperature;
(2) shred: the scopolia acutangula of oven dry is cut into the long segment of 0.5cm;
(3) bottling is soaked: the scopolia acutangula of getting respectively after 10g shreds is loaded in the 150ml triangular flask; Wherein: one bottle of adding 100ml 100% acetone extracts, and other one bottle of adding 100ml 100% methyl alcohol extracts; The sample that two kinds of solvents are extracted respectively is placed on and left standstill under the room temperature 72 hours;
(4) concentrate dissolving: the sample leaching liquor after handling under the above-mentioned different condition is filtered respectively, after concentrating under 50 ℃, be the activity extract of scopolia acutangula; The extract acetone solution of acetone, methanol extraction use the dissolve with methanol sample, quantitatively arrive 0.1g/ml, it is standby to put into refrigerator.
2, control experiment
Since plant sample through organic solvent soak to concentrate the back with acetone and dissolve with methanol quantitatively and measure active, so carry out control experiment for reference with the nematode suspension that does not add the scopolia acutangula extract, contains 5% and 10% acetone, 5% and 10% methanol concentration (volume ratio) respectively.
3, the pine wood nematode used of preparation experiment
(Botrytis cinerea) is inoculated into potato culture (potato: 200g with the fungi Botrytis cinerea, glucose: 20g, agar: 18-20g, water: 1000ml) on the flat board, 25 ℃ are cultured to and cover with flat board, insert a medium that has pine wood nematode, are cultured to mycelia under 25 ℃ to disappear, media surface is glossy to be nematode and to cover with flat board, and it is standby to put into 4 ℃ of refrigerators.The medium that will be loaded with nematode during use is chosen, and wraps up medium with lens wiping paper, puts into the clean flat board that fills sterile water nematode is washed out, and the concentration of adjusting nematode suspension is standby at 100/ml.
4, activity determination method
Do active mensuration respectively with the extract of two kinds of concentration: get 1.9ml pine wood nematode suspension and 100ul scopolia acutangula extract (0.1g/ml) and be added in the culture dish that diameter is 6cm, this moment, activity extract concentration was 5mg/ml, behind the mixing culture dish was put under the room temperature gently and left standstill.Under anatomical lens, select 5-7 the visual field at random, regularly observed respectively, count dead borer population (stiff motionless, be aided with mechanical stimulus, find fault as needle point etc.) and bus borer population as standard with nematode in 12 hours, 24 hours, 48 hours and 72 hours; Other gets 1.8ml pine wood nematode suspension and 200ul scopolia acutangula extract (0.1g/ml) is added in the culture dish that diameter is 6cm, this moment, extract concentrations was 10mg/ml, other experimental techniques are identical with the activity determination method of the extract of 5mg/ml concentration, add up the nemic death rate in the different time respectively.Each specimen is established two repetitions, calculates the average lethality rate of specimen to nematode, just evaluates the virulence of sample to nematode with average lethality rate.Do respectively simultaneously and contain 5% and the control experiment of the nematode suspension of the acetone of 10% two kind of concentration and methyl alcohol (volume ratio).Nemic death rate calculates by following formula:
Lethality %=(observed verge of death borer population/observed bus borer population) * 100%
Activity extract of the present invention has advantages such as preparation is easy, high-efficiency low-toxicity, can be used as the nematicide biopesticide preparation of preparation and uses.
Embodiment:
The scopolia acutangula that present embodiment uses is picked up from the Kunming Inst. of Botany, Chinese Academy of Sciences botanical garden.
Embodiment one:
The furnace drying method oven dry that the scopolia acutangula sample is described through the foregoing invention content part.With 10g length is that the scopolia acutangula oven dry sample of 0.5cm segment is packed in the 150ml triangular flask, 100% acetone soaks, left standstill under the room temperature 72 hours, after the soak filtration, be concentrated into weight not being higher than under 50 ℃ the temperature, weighing, use the acetone solution sample, quantitatively to 0.1g/ml, do the cytotoxicity of the extract of 5mg/ml concentration and 10mg/ml concentration according to above-mentioned experimental technique to pine wood nematode, and with the acetone that contains respective concentration and the nematode suspension that do not add the scopolia acutangula extract in contrast, the results are shown in Table 1.
Table 1 acetone extract is to the lethality rate (%) of pine wood nematode
| 12 hours time | 24 hours time | 48 hours time | 72 hours time | |
| Extract (5mg/ml) | 24.1 | 49.3 | 72.4 | 83.1 |
| Extract (10mg/ml) | 40.7 | 64.1 | 80.3 | 97.2 |
| The contrast of 5% acetone | 1.12 | 2.86 | 4.32 | 6.17 |
| The contrast of 10% acetone | 4.65 | 7.34 | 9.38 | 12.6 |
| The clear water contrast | 1.28 | 3.24 | 4.11 | 6.32 |
The result shows: scopolia acutangula activity extract of the present invention has higher toxic effect to pine wood nematode, its acetone extract has strong inhibitory action to nematode, reaches 97.2% (10mg/ml extract concentrations) and 83.1% (5mg/ml extract concentrations) in 72 hours.To as directed, acetone has certain influence to pine wood nematode, but lethality rate significantly is lower than the scopolia acutangula activity extract under identical time, same concentrations condition, illustrates that activity extract of the present invention has stronger toxic effect to pine wood nematode.
Embodiment two:
Substantially with embodiment one, difference is that the organic solvent that is used to extract is 100% methyl alcohol.The 150ml triangular flask 10g scopolia acutangula oven dry sample of packing into, 100ml methyl alcohol soaks, and leaves standstill 72h under the room temperature, filtering and concentrating quantitatively (0.1g/ml) methanolic extract of obtaining after handling as specimen.Do the cytotoxicity of the extract of 5mg/ml concentration and 10mg/ml concentration according to above-mentioned experimental technique, and with the methyl alcohol that contains respective concentration and the nematode suspension that do not add the scopolia acutangula extract in contrast, activity the results are shown in Table 2 to pine wood nematode.
Table 2 methanolic extract is to the lethality rate (%) of pine wood nematode
| 12 hours time | 24 hours time | 48 hours time | 72 hours time | |
| Extract (5mg/ml) | 22.7 | 54.2 | 68.7 | 81.6 |
| Extract (10mg/ml) | 38.2 | 61.8 | 79.2 | 96.7 |
| The contrast of 5% methyl alcohol | 1.58 | 3.56 | 7.42 | 9.21 |
| The contrast of 10% methyl alcohol | 7.62 | 11.4 | 13.7 | 15.8 |
| The clear water contrast | 1.28 | 3.24 | 4.11 | 6.32 |
The result shows: scopolia acutangula activity extract of the present invention has higher toxic action to pine wood nematode, its methanolic extract has stronger inhibitory action to nematode, reaches 96.7% (10mg/ml extract concentrations) and 81.6% (5mg/ml extract concentrations) in 72 hours.To as directed, methyl alcohol has certain influence to pine wood nematode, but lethality rate significantly is lower than the scopolia acutangula activity extract under identical time, same concentrations condition, illustrates that activity extract of the present invention has stronger toxic action to pine wood nematode.
Claims (2)
1, a kind of active matter that extracts from scopolia acutangula is characterized in that this active matter obtains through following steps:
A. put into baking oven after the scopolia acutangula complete stool that collects being washed with running water, baking is 10 hours under 50 ℃ of temperature;
B. after will toasting scopolia acutangula be cut into the long segment of 0.5cm;
The scopolia acutangula of c. getting respectively after 10g shreds is loaded in the 150ml triangular flask; Wherein: one bottle of adding 100ml 100% acetone extracts, and other one bottle of adding 100ml100% methyl alcohol extracts; The sample that two kinds of solvents are extracted respectively is placed on and left standstill under the room temperature 72 hours;
D. the sample leaching liquor after handling under the above-mentioned different condition is filtered respectively, after concentrating under 50 ℃, be the activity extract of scopolia acutangula; The extract acetone solution of acetone, methanol extraction use the dissolve with methanol sample, quantitatively arrive 0.1g/ml, it is standby to put into refrigerator.
2, the application of the described active matter that extracts from scopolia acutangula of claim 1 is characterized in that the application of this active matter as preparation control pine wood nematode preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100589788A CN101385471A (en) | 2008-09-27 | 2008-09-27 | Active material extracted from Acutangular Anisodus root and uses thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100589788A CN101385471A (en) | 2008-09-27 | 2008-09-27 | Active material extracted from Acutangular Anisodus root and uses thereof |
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| CN101385471A true CN101385471A (en) | 2009-03-18 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276342A (en) * | 2021-12-29 | 2022-04-05 | 锦州拾正生物科技有限公司 | Method for separating and purifying anisodamine monomer from plant extract |
| CN114478515A (en) * | 2022-02-22 | 2022-05-13 | 锦州拾正生物科技有限公司 | Method for separating and purifying anisodine from acutangular anisodine plants |
| CN114507228A (en) * | 2022-02-15 | 2022-05-17 | 锦州拾正生物科技有限公司 | Method for separating and purifying scopolamine monomer from acutangular anisodus root plants |
-
2008
- 2008-09-27 CN CNA2008100589788A patent/CN101385471A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276342A (en) * | 2021-12-29 | 2022-04-05 | 锦州拾正生物科技有限公司 | Method for separating and purifying anisodamine monomer from plant extract |
| CN114507228A (en) * | 2022-02-15 | 2022-05-17 | 锦州拾正生物科技有限公司 | Method for separating and purifying scopolamine monomer from acutangular anisodus root plants |
| CN114478515A (en) * | 2022-02-22 | 2022-05-13 | 锦州拾正生物科技有限公司 | Method for separating and purifying anisodine from acutangular anisodine plants |
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