阅读说明：本技术 南木香在生物防治方面的应用及其生物防治药剂与制备方法 (Application of costus root in biological control, biological control medicament and preparation method thereof ) 是由 刘畅 刘志龙 张治科 刘晓丽 陈宏灏 吴晓燕 魏淑花 黄文广 宋双 王颖 王芳 于 2021-09-22 设计创作，主要内容包括：本发明“南木香AristolochiayunnanensisFranch.在生物防治方面的应用及其生物防治药剂与制备方法”属于生物防治领域。本发明提供南木香AristolochiayunnanensisFranch.在生物防治领域的用途,并基于该用途,本发明进一步提供一种生物防治药剂,其包括活性成分；所述活性成分包括南木香精油。利用本发明的南木香AristolochiayunnanensisFranch.新用途或采用本发明的杀虫剂/杀虫方法可高效杀灭西花蓟马或木虱,且南木香为野生植物,分布范围广泛,简单易得,成本低廉,适于大规模推广应用。(The invention relates to application of Aristolochia yunnanensis Franch, a biological control medicament and a preparation method thereof, belonging to the field of biological control. The invention provides the application of Aristolochia yunnanensis Franch, which belongs to the field of biological control, and based on the application, the invention further provides a biological control medicament which comprises active ingredients; the active ingredient comprises essential oil of costus root. The Aristolochia hancei can be used for efficiently killing Frankliniella occidentalis or psyllid by utilizing the new application or the pesticide/pesticide method, and the Aristolochia hancei is a wild plant, has wide distribution range, is simple and easy to obtain, has low cost and is suitable for large-scale popularization and application.)

1. Application of Aristolochiayunannensis Franch in biological control is provided.

2. Use of aristolochia yunnanensis franch in biocontrol according to claim 1, wherein biocontrol means insecticidal;

preferably, the biological control refers to killing insects by using the essential oil of costus root;

preferably, the insect is selected from psyllid or thrips occidentalis.

3. Use of Aristolochia yunnanensis Franch of Aristolochia according to claim 2 for biological control, wherein said Aristolochia manshurica essential oil is an essential oil extracted from the root of the plant of Aristolochia yunnanensis;

preferably, the active ingredients of the costus root essential oil comprise: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

4. Use of aristolochia yunnanensis franch. according to claim 2, wherein the insecticidal means is selected from fumigation or contact killing;

preferably, the insecticidal semilethal concentration of the costus root essential oil is 0.30mg/L or 0.97 mu g/head.

5. A biological control agent, comprising: an active ingredient; the active ingredient comprises essential oil of costus root.

6. The biocontrol agent of claim 5, wherein said essential oil of Aristolochia debilis comprises as active ingredients: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

7. The biocontrol agent of claim 5 or 6, further comprising: pharmaceutically acceptable adjuvants;

the pharmaceutically acceptable auxiliary materials are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, permeation promoters, pH regulators, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants;

preferably, the biological control agent is an insecticide.

8. A method for preparing biological control medicament is characterized in that the common aucklandia root is extracted with essential oil.

9. The method for producing a biocontrol agent as described in claim 8, wherein said Aristolochia manshurica refers to the root of Aristolochia debilis;

preferably, the essential oil extraction refers to extraction after steam distillation.

10. The process for the preparation of a biocontrol agent as claimed in claim 9 wherein said steam distillation is carried out at an extraction temperature of 70-120 ℃, preferably 100 ℃; the extraction time is 2-8h, preferably 6 h;

preferably, the extracting agent is n-hexane, and the extracting time is 0.2-1h, preferably 0.5 h;

preferably, the n-hexane layer after extraction is dried and concentrated; preferably, the drying refers to removing water from the n-hexane layer with anhydrous sodium sulfate; the concentration refers to that the dried product is subjected to rotary evaporation by using a rotary evaporator to remove n-hexane to obtain the elecampane essential oil;

preferably, the roots of Yunnan Ma Dolichos plants are naturally dried in the shade and crushed before the extraction by the steam distillation method;

preferably, the steam distillation extraction refers to: and (3) putting the roots of the Yunnan aristolochia plants dried and crushed in the shade into a volatile oil extractor for extraction.

Technical Field

The invention belongs to the field of biological control, and particularly relates to application of AristolochiayuanyunnanensissFranch to Aristolochia manshuriensis in biological control, a biological control medicament and a preparation method thereof.

Background

Psyllids are an important group in the homoptera of the Insecta class. It is also a generic term for the homoptera psyllidae family. In the case of metamorphosis insects, individuals develop through the stages of ova, nymphs and adults. The mouth organ is of piercing-sucking type. The adult is small in size and has active energy jumping. If the body is round and flat, waxy secretions cover the body surface. The plant is mainly harmful to woody plants, and important plants include pear psylla harmful to pear trees, mulberry psylla harmful to mulberry trees and the like.

Frankliniella occidentalis is a group of animals of the family thrips, Thripidae. Also known as alfalfa thrips, belongs to the order Thysanoptera. As many as 500 or more host plants are known for the feeding habit of Frankliniella occidentalis. The insect was native to north america, was first discovered in 1955 in hawaii, and was once the most common thrips in california in the united states.

The south costus root is a Chinese medicament prepared from roots, rhizomes and vines of aristolochia yunnanensis franch, has pungent and warm nature and mainly treats the following effects: warming middle energizer, regulating qi-flowing, relieving pain, resolving food stagnation, relaxing muscles and tendons, and activating collaterals; it is indicated for gastritis, abdominal distention, abdominal pain, rheumatic bone pain, etc.

However, the application of the costus root in the field of biological control is not reported in the field, and the application of the costus root in controlling psyllids or frankliniella occidentalis is a blank.

Disclosure of Invention

In order to fill the blank of the prior art in the field, the invention provides application of aristolochiayuanyunnanensissFranch, a biological control medicament and a preparation method thereof.

The technical scheme of the invention is as follows:

application of Aristolochiayunannensis Franch in biological control is provided.

Biological control refers to disinsection;

preferably, the biological control refers to killing insects by using the essential oil of costus root;

preferably, the insect is selected from psyllid or thrips occidentalis.

The Aristolochia nankinensis essential oil refers to essential oil extracted from roots of Yunnan Aristolochia nankinensis plants;

preferably, the active ingredients of the costus root essential oil comprise: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

The insect killing mode is selected from fumigation or contact killing;

preferably, the insecticidal semilethal concentration of the costus root essential oil is 0.30mg/L or 0.97 mu g/head.

A biological control agent, comprising: an active ingredient; the active ingredient comprises essential oil of costus root.

The active ingredients of the costus root essential oil comprise: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

The biological control agent further comprises: pharmaceutically acceptable adjuvants;

the pharmaceutically acceptable auxiliary materials are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, permeation promoters, pH regulators, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants;

preferably, the biological control agent is an insecticide.

A method for preparing biological control medicament is characterized in that the common aucklandia root is extracted with essential oil.

The Aristolochia debilis refers to the root of the Yunnan Madouling plant;

preferably, the essential oil extraction refers to extraction after steam distillation.

The extraction temperature of the steam distillation method is 70-120 ℃, preferably 100 ℃; the extraction time is 2-8h, preferably 6 h;

preferably, the extracting agent is n-hexane, and the extracting time is 0.2-1h, preferably 0.5 h;

preferably, the n-hexane layer after extraction is dried and concentrated; preferably, the drying refers to removing water from the n-hexane layer with anhydrous sodium sulfate; the concentration refers to that the dried product is subjected to rotary evaporation by using a rotary evaporator to remove n-hexane to obtain the elecampane essential oil;

preferably, the roots of Yunnan Ma Dolichos plants are naturally dried in the shade and crushed before the extraction by the steam distillation method;

preferably, the steam distillation extraction refers to: and (3) putting the roots of the Yunnan aristolochia plants dried and crushed in the shade into a volatile oil extractor for extraction.

The invention provides and claims a new application of AristolochiayuanyunnanensissFranch, which is realized by fumigating or contact killing of Siphonothrips cichorii or psyllid with essential oil. The invention also extracts and identifies the active insecticidal components in the AristolochiayuanyunnanensissFranch essential oil for the first time, which comprises the following components: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifoln-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene, and the like. Meanwhile, the invention proves the high lethal efficiency of the AristolochiayannensisFranch essential oil on Franch of Franch frankliniella occidentalis or psyllid through a lethal test, and the half lethal concentration of the Aristolochia yenne essential oil is obviously lower than that of the conventional insecticide. The new application of the Aristolochia yunnanensis Franch of the southern costustoot or the pesticide/insecticidal method of the invention can efficiently kill the Frankliniella occidentalis and the psyllids, and the corresponding Yunnan aristolochia of the southern costustoot is a wild plant, has wide distribution range, is simple and easy to obtain, has low cost and is suitable for large-scale popularization and application.

Detailed Description

The following detailed description of the present invention will be made in conjunction with specific examples and experimental examples, but the scope of the present invention is not limited thereto.

Sources of biological material

The southern costus root of the invention: plants of aristolochia yunnanensis franch, yunnan were purchased from yunnan drug net, yuntang.

Group 1 example, New use of Aristolochia Kamtschatica for biological control

The present group of embodiments provides the use of aristolochia yunnanensis franch.

In some embodiments, biological control refers to killing pests;

preferably, the biological control refers to killing insects by using the essential oil of costus root;

preferably, the insect is selected from psyllid or thrips occidentalis.

In a specific embodiment, the aristolochia manshuriensis essential oil refers to an essential oil extracted from roots of aristolochia manshuriensis plants;

preferably, the active ingredients of the costus root essential oil comprise: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

In other embodiments, the insecticidal regimen is selected from fumigation or contact killing;

preferably, the insecticidal semilethal concentration of the costus root essential oil is 0.30mg/L or 0.97 mu g/head.

Based on the teachings of the present invention, one skilled in the art can use aristolochia yunnanensis franch, or any part of the plant thereof, or an extract thereof, for biocontrol applications including, but not limited to: insecticidal, acaricidal, attractant for animals (including but not limited to insects), repellant, and the like; such insects include, but are not limited to, thrips occidentalis, psyllid, and the like; any action of using aristolochia yunnanensis franch, or derivative products derived from aristolochia yunnanensis franch (e.g., extracts, extracted essential oils, volatile oils, formulations containing any part or tissue of aristolochia yunnanensis franch) for biological control falls within the scope of the present invention.

Group 2 examples of biocontrol agents of the invention

The present group of embodiments provides a biological control agent. All embodiments of this group share the following common features: the biocontrol agent comprises: an active ingredient; the active ingredient comprises essential oil of costus root.

In a specific embodiment, the active ingredients of the costus root essential oil comprise: 9-isopropylbicyclo [ 6.1.0 ] nonaene, (3R) -1-4,4a,5,6,7, 8-hexahydro-4, 4 a-dimethyl-6- (1-methylvinyl) -2(3H) -naphthalenone, 3,7, 7-trimethyl-11-methylene- (6R) -spiro [ 5.5 ] undec-2-ene, methylene-3-prop-1-en-2-yl-cyclohexane, bornyl acetate, (1R,4aR,5R,9aS) -4,5,9, 9-tetramethyl-1, 4a,5,6,7,8,9,9 a-octahydro-1, 5-ring, (-) -4-terpineol, Eudesmol, (-) -isolongifolen-9-one, perillaldehyde, 2-sagittanol, 1,7, 7-trimethylbicyclo [ 2.2.1 ] heptan-2-yl formate, eucalyptol, (1R) -3 alpha-vinyl-3-methyl-2 beta- (1-methylvinyl) -6 beta-isopropylcyclohexane-1 beta-ol, 8-dimethyl-9-methylene-1, 5-cycloundecadiene.

In a further embodiment, the biocontrol agent further comprises: pharmaceutically acceptable adjuvants;

the pharmaceutically acceptable auxiliary materials are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integration agents, permeation promoters, pH regulators, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants;

preferably, the biological control agent is an insecticide.

According to the description of the present invention, the skilled in the art can select and adjust the above-mentioned auxiliary materials according to the needs of practical production and application, for example, the dosage form of the biological control agent of the present invention can be further prepared as follows: liquid, spray, powder, tablet and other conventional dosage forms, and the required supplementary material is selected and the supplementary material amount is prepared based on the conventional dosage forms.

Group 3 examples, Process for the preparation of the biocontrol agents of the present invention

The present group of embodiments provides a method of preparing a biological control agent. All embodiments of this group share the following common features: the preparation method of the biological control medicament comprises the following steps: extracting essential oil from radix aucklandiae.

In specific embodiments, the aristolochia manshurica refers to the root of the yunnan aristolochia plant;

preferably, the essential oil extraction refers to extraction after steam distillation.

In some embodiments, the steam distillation process has an extraction temperature of 70-120 ℃, preferably 100 ℃; the extraction time is 2-8h, preferably 6 h;

preferably, the extracting agent is n-hexane, and the extracting time is 0.2-1h, preferably 0.5 h;

preferably, the n-hexane layer after extraction is dried and concentrated; preferably, the drying refers to removing water from the n-hexane layer with anhydrous sodium sulfate; the concentration refers to that the dried product is subjected to rotary evaporation by using a rotary evaporator to remove n-hexane to obtain the elecampane essential oil;

preferably, the roots of Yunnan Ma Dolichos plants are naturally dried in the shade and crushed before the extraction by the steam distillation method;

preferably, the steam distillation extraction refers to: and (3) putting the roots of the Yunnan aristolochia plants dried and crushed in the shade into a volatile oil extractor for extraction.

Experimental example, concrete preparation process, component identification and effect verification of biological control medicament

1. Preparation of plant essential oil

The method comprises the steps of taking roots of a costus root plant, naturally drying in the shade, crushing, weighing, placing in a volatile oil extractor, extracting by adopting a steam distillation method for 6 hours generally, extracting by adopting n-hexane to obtain an n-hexane layer, adding anhydrous sodium sulfate to remove water, concentrating, measuring the volume, placing the volatile oil in a dry glass bottle, and placing in a refrigerator at 4 ℃ for storage.

2. GC-MS testing

The volatile oil sample adopts GC-MS to analyze the component composition of the sample, and the test conditions are as follows: GCMS-QP2010 plus (Shimadzu); a chromatographic column: DB-5MS (60m 0.25um 0.32mm) (Agilent); the heating rate is as follows: 60-10 ℃/min-300-20 min; the split ratio is as follows: 30: 1; constant linear velocity mode: 30 cm/sec; carrier gas: he; ion source temperature: 200 ℃; interface temperature: 300 ℃; sample inlet temperature: 320 ℃; mass scan range: m/z 29-650 sample size: 1 mu L of the solution; sample pretreatment: filtering the sample by a filter head containing a 0.22 mu m organic filter membrane of a disposable syringe; and (3) searching a spectrum library: the Nist 2017 standard library.

The compositions of the costus root essential oil are shown in the following table 1 after detection:

TABLE 1

The total percentage of all the components identified in table 1 above is less than 100% because unknown components still exist in the essential oil of aristochip ayunnanensis franch, which may be new compounds and cannot be identified by the existing GC-MS detection method.

4. Determination of fumigating activity of volatile oil of Aristolochia nankinensis on Frankliniella occidentalis

A fumigation method is adopted, 10 adults of Frankliniella occidentalis are sucked into a 20mL small bottle, the small bottle is sleeved with gauze and placed into a 250mL wide-mouth bottle, a 1cm multiplied by 6cm filter paper strip is fixed in the bottle cap, the filter paper strip with 20 microliter of liquid medicine is aired for 10-20s, the bottle cap is covered after the solvent is volatilized, the filter paper strip is suspended in the center of the wide-mouth bottle, and the wide-mouth bottle cap is wrapped by a sealing film. After treatment, the jar is placed in a plant incubator, and the temperature (25 +/-0.5) DEG C, the relative humidity 50%, the illumination L: d is 16h:8h of illumination climate box. The negative and positive controls are negative and positive, respectively, with no volatile oil solvent and neem oil added. Repeating each treatment for 6 times, repeating each time for 10 thrips, checking death and survival condition of thrips in each treatment 24 hr later, calculating death rate, and calculating semilethal concentration LC by DPS₅₀。

5. Determination of touch killing activity of volatile oil of common aucklandia root to psyllid

The contact activity was determined by the microdroplet method. The test insects are anesthetized by freezing for 10min, 10 psyllids/tube are dripped on the forethorax and the backboard of the insect body by adopting a dripping method, and 2 mu L of the test insects are dripped into each head. And (3) placing the treated psyllids in a centrifuge tube filled with the fresh and tender branches and buds of the Chinese wolfberry, and sealing the centrifuge tube by using a gauze. After treatment, the centrifuge tube is placed in an intelligent artificial illumination climate box, and the illumination climate box is set at a temperature (25 +/-0.5 ℃), has a relative humidity of 60% and has an illumination L: D of 16h:8 h. The treatment without drug was used as a blank control, the group added with neem oil was used as a positive control, and the group added with the solution of essential oil of Aristolochia debilis was used as an experimental group. Repeating each treatment for 6 times, repeating 10 psyllids, checking death and survival condition of each treated psyllid 24h after treatment, calculating death rate, and calculating semi-lethal concentration LC by DPS₅₀。

The comparative results are shown in table 2 below:

TABLE 2