阅读说明：本技术 马兰在生物防治领域的用途及其生防药剂、制备方法及杀虫方法 (Application of Kalimeris indica in biological control field, biocontrol agent, preparation method and insecticidal method thereof ) 是由 刘畅 马建华 张治科 白小军 周涛 许泽华 王颖 李百云 郭鑫年 王贵荣 于 2021-08-17 设计创作，主要内容包括：本发明“马兰Aster indicus在生物防治领域的用途及其生防药剂、制备方法及杀虫方法”属于生物防治领域。本发明提供马兰Aster indicus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；所述活性成分包括马兰精油。本发明还提供一种生物防治药剂的制备方法,其特征是,以马兰精油做为所述生物防治药剂的活性成分。本发明还提供一种杀虫方法,其特征是采用马兰精油杀虫。利用本发明的马兰Aster indicus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、棉蚜或木虱,且马兰为野生植物,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 discloses application of Indian kalimeris herb in the field of biological control, a biological control medicament, a preparation method and an insecticidal method thereof, and belongs to the field of biological control. The invention provides the use of the Aster indicus of Kalimeris in the field of biological control, and based on the use, the invention further provides a biological control medicament which comprises active ingredients; the active ingredient comprises Kalimeris essential oil. The invention also provides a preparation method of the biological control medicament, which is characterized in that the kalimeris indica essential oil is used as an active ingredient of the biological control medicament. The invention also provides a disinsection method which is characterized in that the kalimeris indica essential oil is adopted to kill the pests. The novel application of the Indian kalimeris Aster indicus or the pesticide/insecticidal method can efficiently kill Frankliniella occidentalis, Aphis gossypii or psyllid, and the Indian kalimeris 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 Aster indicus in the field of biological control.

2. Use of the Indian orchid Aster indicus according to claim 1, characterized by the use of Indian orchid essential oil in the field of biological control;

preferably, the kalimeris indica essential oil refers to an essential oil extracted from the overground part of kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone.

3. Use of blue Aster indicus according to claim 1 or 2 for biocontrol, wherein biocontrol refers to disinfestation;

the insect is preferably an agricultural pest;

preferably, the agricultural pest is selected from: cotton aphid, thrips occidentalis, or psyllid;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the pesticidal semilethal concentration of said Kalimeris indica essential oil is between 0.53 and 10.11mg/mL, preferably 0.53mg/mL, 0.96mg/mL, or 10.11 mg/mL.

4. A biological control agent characterized by comprising an active ingredient; the active ingredient comprises Kalimeris essential oil.

5. The biocontrol agent of claim 4, wherein said Kalimeris indica essential oil is an essential oil extracted from the aerial parts of Kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the dosage form of the biocontrol agent is selected from biopesticides, insecticides; the insect is preferably cotton aphid, frankliniella occidentalis or psylla;

preferably, the biocontrol agent further comprises a pharmaceutically acceptable adjuvant.

6. A preparation method of a biological control medicament is characterized in that Kalimeris indica essential oil is used as an active ingredient of the biological control medicament.

7. The process for the preparation of a biocontrol agent as claimed in claim 7 wherein the above ground parts of kalimeris indica are subjected to essential oil extraction;

preferably, the aerial parts of kalimeris indica are extracted with an extractant; preferably, the extracting agent is n-hexane, and the extracting time is 4-12h, preferably 6 h;

preferably, the aerial parts of kalimeris indica are dried in the shade and crushed before extraction; the extraction refers to placing the overground part of the kalimeris indica in a volatile oil extractor for extraction;

preferably, the n-hexane layer obtained after extraction is dried and concentrated; preferably, the drying refers to adding anhydrous sodium sulfate into the n-hexane layer to remove water; preferably, the concentration refers to that the dried product is evaporated by a rotary evaporator to remove n-hexane to obtain the kalimeris indica essential oil.

8. An insecticidal method is characterized in that kalimeris indica essential oil is adopted to kill insects.

9. A method as claimed in claim 9, wherein said insects are selected from the group consisting of: cotton aphid, thrips occidentalis, or psyllid;

the Kalimeris essential oil is preferably an essential oil extracted from the aerial parts of Kalimeris;

preferably, the kalimeris indica essential oil comprises the following compounds with insecticidal activity: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the pesticide refers to killing with a kalimeris indica essential oil solution; preferably, the maranta essential oil solution has a semi-lethal concentration of 0.53-10.11mg/ml, preferably 0.53mg/ml, 0.96mg/ml, or 10.11 mg/ml.

Technical Field

The invention belongs to the technical field of biological control, and particularly relates to application of Indian kalimeris herb Aster indicus in the field of biological control, a biological control medicament, a preparation method and an insecticidal method thereof.

Background

The Aster indicus of the Indian Aster genus is the Aster plant of Asteraceae family of Antiospermae of angiosperm phylum, and the whole herb or root thereof can be used as a medicine, and is recorded in the book of medicine such as national Chinese herbal medicine compilation and Chinese medicine dictionary. The plant is mostly grown in low mountainous areas, flat dams or wet areas of hills. Distributed in each province of China.

Malan is pungent, bitter and cold in nature and mainly treats functions: clear heat and remove toxicity, dissipate blood stasis and stop bleeding, and remove food retention. Can be used for treating common cold, fever, cough, acute pharyngitis, tonsillitis, epidemic parotitis, infectious hepatitis, gastric ulcer, duodenal ulcer, infantile malnutrition, enteritis, dysentery, hematemesis, metrorrhagia, and menoxenia; it is used externally to treat sore, furuncle, swelling and pain, mastitis, traumatic hemorrhage.

However, the use of kalimeris has not been reported in the field of biological control.

Disclosure of Invention

Based on the blank existing in the prior art in the field, the invention firstly provides the new application of the Indian kalimeris herb Aster indicus in the field of biological control, and provides a biological control medicament taking the Indian kalimeris herb as a raw material and a preparation method thereof based on the new application.

The technical content of the invention is as follows:

application of Aster indicus in the field of biological control.

The application of the Indian kalimeris herb Aster indicus in the aspect of biological control refers to the application of kalimeris herb essential oil in the field of biological control;

preferably, the kalimeris indica essential oil refers to an essential oil extracted from the overground part of kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone.

The biological control refers to disinsection;

the insect is preferably an agricultural pest;

preferably, the agricultural pest is selected from: cotton aphid, thrips occidentalis, or psyllid;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the pesticidal semilethal concentration of said Kalimeris indica essential oil is between 0.53 and 10.11mg/mL, preferably 0.53mg/mL, 0.96mg/mL, or 10.11 mg/mL.

A biological control agent characterized by comprising an active ingredient; the active ingredient comprises Kalimeris essential oil.

The Kalimeris indica essential oil is extracted from aerial parts of Kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the dosage form of the biocontrol agent is selected from biopesticides, insecticides; the insect is preferably cotton aphid, frankliniella occidentalis or psylla;

preferably, the biocontrol agent further comprises a pharmaceutically acceptable adjuvant.

A preparation method of a biological control medicament is characterized in that Kalimeris indica essential oil is used as an active ingredient of the biological control medicament.

The preparation method of the biological control medicament comprises the steps of extracting essential oil from the overground part of the kalimeris indica;

preferably, the aerial parts of kalimeris indica are extracted with an extractant; preferably, the extracting agent is n-hexane, and the extracting time is 4-12h, preferably 6 h;

preferably, the aerial parts of kalimeris indica are dried in the shade and crushed before extraction; the extraction refers to placing the overground part of the kalimeris indica in a volatile oil extractor for extraction;

preferably, the n-hexane layer obtained after extraction is dried and concentrated; preferably, the drying refers to adding anhydrous sodium sulfate into the n-hexane layer to remove water; preferably, the concentration refers to that the dried product is evaporated by a rotary evaporator to remove n-hexane to obtain the kalimeris indica essential oil.

An insecticidal method is characterized in that kalimeris indica essential oil is adopted to kill insects.

The insect is selected from: cotton aphid, thrips occidentalis, or psyllid;

the Kalimeris essential oil is preferably an essential oil extracted from the aerial parts of Kalimeris;

preferably, the kalimeris indica essential oil comprises the following compounds with insecticidal activity: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the pesticide refers to killing with a kalimeris indica essential oil solution; preferably, the maranta essential oil solution has a semi-lethal concentration of 0.53-10.11mg/ml, preferably 0.53mg/ml, 0.96mg/ml, or 10.11 mg/ml.

The invention discovers and requests to protect the new application of the Aster indicus in the aspect of biological control for the first time, and the new application is mainly realized by fumigating or contact killing of frankliniella occidentalis, cotton aphid or psylla. The invention also extracts and identifies the active insecticidal components in the Indian kalimeris essential oil for the first time, wherein the active insecticidal components comprise linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaiene, eucalyptol, epoxidized lupinene II, arylturmerone and the like. Meanwhile, the lethal test proves that the high lethal efficiency of the Aster indicus essential oil on Frankliniella occidentalis, Aphis gossypii or psylla is high, and the half lethal concentration of the Aster indicus essential oil is obviously lower than that of the conventional pesticide. The novel application of the Indian kalimeris Aster indicus or the pesticide/insecticidal method can efficiently kill Frankliniella occidentalis, Aphis gossypii or psyllid, and the Indian kalimeris 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.

Group 1 examples, New use of Marantan of the invention for biological control

The embodiment of the group provides application of the Indian kalimeris in the field of biological control.

In a specific embodiment, the use refers to the use of the kalimeris indica essential oil in the field of biological control;

preferably, the kalimeris indica essential oil refers to an essential oil extracted from the overground part of kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone.

The compounds listed above are components of the kalimeris indica essential oil accounting for 2% by mass or more, and components accounting for 2% by mass or more are listed as the insecticidal active ingredients of the kalimeris indica essential oil of the present invention on the assumption that the main components are active ingredients, but the components of the kalimeris indica essential oil or the insecticidal active ingredients are not limited to the above compounds, and all compounds of the kalimeris indica essential oil detectable under the existing detection precision conditions are specifically shown in experimental example table 1.

In some embodiments, the biological control refers to disinsection;

the insect is preferably an agricultural pest;

preferably, the agricultural pest is selected from: cotton aphid, thrips occidentalis, or psyllid;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the malan essential oil has an insecticidal semilethal concentration of 0.53-10.11mg/ml, preferably 0.53mg/ml, 0.96mg/ml, or 10.11 mg/ml.

Based on the teachings of the present invention, one skilled in the art can use kalimeris indica or any part of its plant, or an extract thereof, for biocontrol uses 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, cotton aphids, thrips occidentalis, psyllids, and the like; any action of using kalimeris indica, or derived products derived from kalimeris indica (e.g., extracts, extractives, extracted essential oils, formulations containing any part or tissue of kalimeris indica) for biological control is 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 biological control agent comprises an active ingredient; the active ingredient comprises Kalimeris essential oil.

In some embodiments, the kalimeris indica essential oil refers to an essential oil extracted from the aerial parts of kalimeris indica;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the dosage form of the biocontrol agent is selected from biopesticides, insecticides; the insect is preferably cotton aphid, frankliniella occidentalis or psylla;

preferably, the biocontrol agent further comprises a pharmaceutically acceptable adjuvant;

preferably, the pharmaceutically acceptable excipients include: 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 enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, antifoaming agents, thickeners, encapsulation agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retardants.

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 Indian kalimeris herb essential oil is used as an active ingredient of the biological control medicament.

In a specific embodiment, the above-ground parts of kalimeris indica are subjected to essential oil extraction;

preferably, the aerial parts of kalimeris indica are extracted with an extractant; preferably, the extracting agent is n-hexane, and the extracting time is 4-12h, preferably 6 h;

preferably, the aerial parts of kalimeris indica are dried in the shade and crushed before extraction; the extraction refers to placing the overground part of the kalimeris indica in a volatile oil extractor for extraction;

preferably, the n-hexane layer obtained after extraction is dried and concentrated; preferably, the drying refers to adding anhydrous sodium sulfate into the n-hexane layer to remove water; preferably, the concentration refers to that the dried product is evaporated by a rotary evaporator to remove n-hexane to obtain the kalimeris indica essential oil.

Group 4 examples, pesticidal methods of the invention

The present group of embodiments provides a method of killing insects. All embodiments of this group share the following common features: the kalimeris indica essential oil is adopted to kill pests.

In some specific embodiments, the insect is selected from: thrips occidentalis, psyllid, or aphis gossypii;

the Kalimeris essential oil is preferably an essential oil extracted from the aerial parts of Kalimeris;

preferably, the kalimeris indica essential oil comprises the following compounds with insecticidal activity: linalool, a, 4-trimethyl-1-cyclohexene-1-methanol, (-) -caryophyllene oxide, n-pentadecanoic acid, phytol, alpha-guaialene, eucalyptol, epoxidized lupinene II and ar-turmerone;

preferably, the biological control active ingredients of the kalimeris indica essential oil comprise the following compounds in percentage by weight: 20.415% of linalool, 15.2095% of a, a, 4-trimethyl-1-cyclohexene-1-methanol, 14.7719% of (-) -eugenol oxide, 11.3685% of n-pentadecanoic acid, 5.0026% of phytol, 3.0973% of alpha-guaiene, 2.9425% of eucalyptol, 2.4088% of epoxidized lupinene II and 2.1337% of arylturmerone;

preferably, the insecticidal mode is contact killing or fumigating killing;

preferably, the pesticide refers to killing with a kalimeris indica essential oil solution; preferably, the maranta essential oil solution has a semi-lethal concentration of 0.53-10.11mg/ml, preferably 0.53mg/ml, 0.96mg/ml, or 10.11 mg/ml.

Wherein the semilethal concentration of the herba Kalimeridis essential oil solution on cotton aphid is 10.11mg/ml, the semilethal concentration of psyllid is 0.96mg/ml, and the semilethal concentration of Frankliniella occidentalis is 0.53 mg/ml.

The herba Kalimeridis essential oil solution is formed by dissolving herba Kalimeridis essential oil in alcohol solvent.

Experimental example, preparation of Kalimeris indica essential oil and verification of effect thereof

1. Preparation of plant essential oil

Drying aerial parts of Kalimeris indica in the shade, pulverizing, weighing, placing in volatile oil extractor, extracting by steam distillation for 6 hr, extracting with n-hexane to obtain n-hexane layer, adding anhydrous sodium sulfate to remove water, concentrating, measuring volume, placing volatile oil in dry glass bottle, and storing in refrigerator at 4 deg.C.

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 components of the kalimeris indica essential oil are detected as shown in the following table 1:

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 Kalimeris essential oil, which may be new compounds that cannot be identified by the existing GC-MS detection method.

3. Determination of contact killing activity of Kalimeris indica volatile oil on cotton aphids

Measuring insecticidal activity by leaf disc method, coating polytetrafluoroethylene on four walls of 12-pore plate, air drying, spreading agar on plate bottom, taking fresh cotton leaf without pesticide, perforating the leaf to obtain circular leaf disc with diameter of 3cm, soaking the leaf in pesticide (i.e. herba Kalimeridis essential oil solution formed by dissolving herba Kalimeridis essential oil in alcohol solvent) for 10s, air drying, placing into pore plate, inoculating 10 fructus Lycii Aphis gossypii, and standing at 25 deg.C with light/dark for 16h/8hAnd the humidity is 60-70%. The negative control and the positive control are respectively the solvent without adding volatile oil and pyrethrin. Repeating each treatment for 6 times, repeating each time for 10 cotton aphids, checking the death number and survival condition of each cotton aphid after 24h, calculating the death rate, and calculating the semi-lethal concentration LC by DPS₅₀。

4. Determination of fumigating activity of Kalimeris occidentalis volatile oil on Frankliniella occidentalis

The insecticidal activity is measured by adopting a fumigation method, 10 heads of adult frankliniella occidentalis are sucked into a 20mL small bottle, the adult frankliniella occidentalis is sleeved with a 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 dropping 20 mu l of liquid medicine (namely a malan essential oil solution formed by dissolving the malan essential oil in an alcohol solvent) 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 control and the positive control are respectively the solvent without adding volatile oil and pyrethrin. 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 contact killing activity of Kalimeris indica volatile oil on psyllids

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 forebreast and back plate of the insect body by adopting a dripping method, and 2 mu L of an Indian kalimeris herb essential oil solution is dripped on each head of the insect body. 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. Negative and positive controls are set with solvent without volatile oil and neem oil. 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