Application of Artemisia sphaerocephala in preventing and controlling agricultural pests, its microcapsule, preparation method and prevention method

Abstract

The present invention, "Application of Artemisia sphaerocephala in biological control, its biological control agent, and preparation method," belongs to the field of biological control. The present invention provides the use of Artemisia sphaerocephala in the field of biological control, and based on this use, the present invention further provides a biological control agent comprising an active ingredient; the active ingredient comprises Artemisia sphaerocephala essential oil. Utilizing the use of Artemisia sphaerocephala in the present invention or employing the insecticide / insecticide method of the present invention can effectively kill cotton aphids, western flower thrips, or wolfberry psyllids. Artemisia sphaerocephala is widely distributed, easily available, and inexpensive, making it suitable for large-scale promotion and application.

Description

Technical Field

[0001] The invention belongs to the technical field of biological control, and relates to the application of Artemisia sphaerocephala in preventing and controlling agricultural pests, and a microcapsule preparation thereof, a preparation method and a control method thereof. Background Art

[0002] Artemisia sphaerocarpa ( Artemisia ordosica Artemisia ordosica, commonly known as Artemisia ordosica, is a plant of the Asteraceae family. The entire plant is used medicinally, often for its anti-inflammatory and swelling properties, chest-relieving and qi-invigorating properties, insecticide, and hemostasis. It can treat rheumatoid arthritis, mumps, scabies, abdominal distension, intestinal obstruction, and urinary retention. It is a major and dominant species of vegetation in sandy areas. Artemisia ordosica is a small shrub, 50-80 cm tall, with deep roots, numerous and thick woody taproots, robust rhizomes, and dense, soft, creeping branches above ground. Previous literature reports indicate that Artemisia ordosica contains trace elements, volatile components, and non-volatile chemical compounds.

[0003] Lycium barbarum psyllid, scientific name: Poratriozasinica Yang et Li. This insect, belonging to the order Hemiptera and family Lycium barbarum, infests a variety of fruit trees, including wolfberries and nightshades. Adults and nymphs insert their mouthparts into leaf tissue on the underside of leaves, sucking sap. This causes yellowing leaves and branches, weakening the tree, inhibiting berry development and quality, and causing branch dryness in spring.

[0004] The western flower thrips is a member of the Thripidae family, Thysanoptera order. It has a varied diet and is known to host over 500 plant species. Native to North America, it was first discovered on the Hawaiian island of Kauai in 1955 and was once the most common thrips in California.

[0005] Cotton Aphid, scientific name: Aphis gossypii Glover Aphids are a species of insect in the Aphididae family, commonly known as the sticky insect. Aphids are a worldwide cotton pest. They occur in all cotton-growing areas of China and are a major pest of cotton seedlings. Their host plants include pomegranates, Sichuan peppercorns, hibiscus, rhamnosus, cotton, and melons.

[0006] Currently, pest control methods primarily include physical and mechanical methods and chemical methods. Physical and mechanical control, which exploits the behavior of pests, involves physically killing insects and mites, degrading their habitat, or mechanically separating them from food to prevent or mitigate pest infestations. Chemical control, which utilizes insecticides, offers rapid, thorough, and broad-spectrum pest control. However, insecticides are toxic to humans and higher animals. In the 1990s, the long-term use of highly toxic chemical pesticides led to over 50,000 poisonings and 5,000 to 7,000 deaths worldwide each year. These pesticides also contribute to the development of pesticide resistance in pests.

[0007] Botanical pesticides are primarily composed of naturally occurring compounds. These active substances are primarily composed of elements such as C, H, and O. Derived from nature, they have developed fixed energy and material circulation metabolic pathways through long evolutionary processes. Therefore, they are less likely to leave residues when applied, are easily degraded, and are less likely to accumulate toxicity in the environment. They are environmentally compatible, maintaining the high quality of agricultural products and meeting people's needs for improving their quality of life and earning foreign exchange through exports. Since China's entry into the WTO, botanical pesticides have helped to support the growing international demand for organic foods (AA grade) and pollution-free products (A grade), avoiding the negative impacts of highly toxic chemical pesticides. Compared to chemical pesticides, botanical pesticides are generally non-toxic, residue-free, and safe for humans and animals. The active ingredients in botanical pesticides are easily broken down into inactive substances that cannot accumulate in pests. Therefore, they are less likely to harm beneficial insects or natural enemies of pests, thus contributing to ecological protection.

[0008] Currently, there is no report in the art that Artemisia sphaerocarpa can be used to kill pests in the field of biological control. Summary of the Invention

[0009] In response to the above-mentioned needs and gaps in the field, the present invention provides a method and agent for controlling pests that are safe and non-toxic to humans and animals, friendly to the environment, and effective.

[0010] The technical solutions claimed in the present invention are as follows:

[0011] Application of Artemisia sphaerocephala in preventing and controlling agricultural pests; the agricultural pests are selected from western flower thrips, wolfberry psyllids or cotton aphids.

[0012] Using Artemisia sphaerocarpa essential oil to control agricultural pests;

[0013] Preferably, the Artemisia sphaerocarpa essential oil is an essential oil extracted from the aerial part of the plant.

[0014] The active ingredients of the black artemisia essential oil include biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cube alcohol, and α-pinene.

[0015] A medicament for preventing and controlling agricultural pests, characterized in that its active ingredient comprises: black artemisia essential oil; the agricultural pests are selected from western flower thrips, wolfberry psyllids, and cotton aphids.

[0016] The black artemisia essential oil is an essential oil extracted from the above-ground part of the plant;

[0017] Preferably, the active ingredients of the Artemisia sphaerocarpa essential oil include: biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cubediol, and α-pinene.

[0018] The dosage form of the biological control agent is selected from the group consisting of: spray, powder, tablet, capsule, and liquid preparation;

[0019] Preferably, the biological control agent further comprises a pharmaceutically acceptable excipient.

[0020] A method for preparing a medicament for preventing and controlling agricultural pests is characterized by comprising: using black artemisia essential oil as an active ingredient of the biological control medicament.

[0021] The black artemisia essential oil is an essential oil extracted from the above-ground part of the plant;

[0022] Preferably, the aerial parts of the Artemisia sphaerocarpa plant are extracted by steam distillation and then extracted;

[0023] Preferably, the extraction temperature of the steam distillation method is 70-120°C, preferably 90°C; the extraction time is 4-12h, preferably 6h;

[0024] Preferably, the extractant is n-hexane, and the extraction time is 2-6 hours, preferably 3 hours;

[0025] Preferably, the extracted n-hexane layer is dried and concentrated;

[0026] Preferably, the drying refers to removing moisture from the n-hexane layer with anhydrous sodium sulfate;

[0027] The concentration refers to removing n-hexane from the dried product by rotary evaporation using a rotary evaporator to obtain black artemisia essential oil;

[0028] Preferably, the above-ground part of the Artemisia sphaerocephala plant is naturally dried in the shade and chopped before extraction by steam distillation;

[0029] Preferably, the steam distillation extraction method comprises: placing the air-dried and chopped above-ground parts of the Artemisia sphaerocephala plant in a round-bottom flask and extracting using an essential oil extractor;

[0030] Preferably, the method for preparing the biological control agent further comprises: adding pharmaceutically acceptable excipients.

[0031] The agent for controlling agricultural pests is selected from a liquid agent and a microcapsule agent; preferably, the liquid agent is an Artemisia sphaerocephala essential oil solution obtained by dissolving Artemisia sphaerocephala essential oil in ethanol; preferably, the microcapsule agent adopts an inclusion complexation method to add a core material solution containing Artemisia sphaerocephala essential oil dropwise to a wall material solution, stir, let stand, filter, remove unreacted essential oil, and then dry;

[0032] The weight ratio of the core material solution to the wall material solution is 1:4-1:8;

[0033] The core material solution is obtained by dissolving black artemisia essential oil in ethanol solution and stirring until it is completely dissolved;

[0034] The wall material solution is obtained by dissolving β-cyclodextrin in deionized water and stirring until completely dissolved;

[0035] The dissolution temperature is 30-60°C, preferably 50°C;

[0036] The cyclodextrin mass concentration is 2.83% w / v; the mass concentration of the obtained microcapsules is measured, and no selection conditions are given.

[0037] The stirring speed is 200-1000 r / min, preferably 500 r / min;

[0038] The stirring time is 1-2 h;

[0039] The standing is standing in a 4°C refrigerator for 24 hours;

[0040] The unreacted essential oil is removed by washing with ethanol;

[0041] The drying temperature is 30-60°C, preferably 40°C;

[0042] The drying time is 3-6 h, preferably 4 h.

[0043] A method for controlling agricultural pests, characterized in that Artemisia sphaerocephala is used to control agricultural pests; the agricultural pests are selected from western flower thrips, wolfberry psyllids, and cotton aphids;

[0044] Preferably, the insecticide is carried out using a solution of Artemisia sphaerocarpa essential oil;

[0045] Preferably, the insecticide method is selected from: contact killing or fumigation killing;

[0046] Preferably, the half-lethal concentration of the Artemisia sphaerocephala essential oil solution for killing cotton aphids is 5.87 mg / mL;

[0047] Preferably, the half lethal concentration of the Artemisia sphaerocephala essential oil solution for killing western flower thrips is 1.57 mg / mL;

[0048] Preferably, the half-lethal concentration of the Artemisia sphaerocephala essential oil solution for killing Lycium barbarum psyllids is 0.81 mg / mL.

[0049] This invention proposes and claims protection for the novel use of Artemisia sphaeroceae in biological control, primarily through the fumigation or contact killing of western flower thrips, wolfberry psyllids, and cotton aphids using black Artemisia sphaeroceae essential oil. The invention also extracts and identifies for the first time the active insecticidal components of black Artemisia sphaeroceae essential oil, which include biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cubediol, and α-pinene. Furthermore, lethality tests confirm the high lethality of black Artemisia sphaeroceae essential oil against western flower thrips, wolfberry psyllids, and cotton aphids. The toxicity of black Artemisia sphaeroceae essential oil against cotton aphids was lower than that of the control agent, pyrethrum extract, but higher than that of the control agent, pyrethrum extract, against wolfberry psyllids. Its toxicity against western flower thrips was higher than that of the control agent, neem oil. The new use of Artemisia sphaerocephala or the insecticide / insecticide method of the present invention can effectively kill western flower thrips, wolfberry psyllids, and cotton aphids. Artemisia sphaerocephala is widely distributed, easily available, and low in cost, making it suitable for large-scale promotion and application. DETAILED DESCRIPTION

[0050] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are only for explanation and illustration and should not be construed as limiting the scope of protection of the present invention.

[0051] Sources and acquisition methods of experimental materials

[0052] Cotton Aphid ( Aphis gossypii ) were inoculated on cotton leaves in the greenhouse, and the plants were subcultured and propagated for three generations before use. The rearing conditions were: temperature (25±1)℃, humidity (40±5)%, L / / D = 16 h / / 8 h.

[0053] Lycium barbarum Psyllid ( Bactericeragobica ) were collected from the pesticide-free wolfberry garden of the Lycium barbarum Research Institute of Ningxia Academy of Agriculture and Forestry Sciences. They were subcultured and propagated for three generations before use. The rearing conditions were: temperature (26±1)℃, humidity (40±5)%, L / / D = 16 h / / 8 h.

[0054] Western flower thrips ( Frankliniella occidentalis ) were collected from the pesticide-free wolfberry garden of the Lycium barbarum Research Institute of the Ningxia Academy of Agricultural and Forestry Sciences and reared with common bean pods for at least three generations in an artificial climate chamber. Rearing conditions were: temperature (25 ± 1)°C, humidity (40 ± 5)%, L / / D (L / / D) = 16 h / / 8 h.

[0055] The applicant declares that it may be distributed to the public for necessary verification tests within twenty years from the date of application.

[0056] Artemisia sphaerocarpa ( Artemisia ordosica ), the collection time was May 28, 2021, and the collection location was Dakouzigou, Yuding Township, Zhongwei City, Ningxia Hui Autonomous Region.

[0057] Black Artemisia sphaeroceae essential oil: The method of the present invention is suitable for extracting only volatile components that can be distilled with steam without being destroyed, do not react with water, and are poorly soluble or insoluble in water. The method of preparing black Artemisia sphaeroceae essential oil of the present invention is as shown in Experimental Example 1.

[0058] Test agents: Pyrethrum extract (25% pyrethrin I + pyrethrin II, Sigma); neem oil (Jiangxi Xuesong Natural Medicinal Oil Co., Ltd.); food-grade β-cyclodextrin (Shandong Tongwang Biotechnology Co., Ltd.); pyrethrin aqueous emulsion (1.5%, Yunnan Nanbao Biotechnology Co., Ltd.)

[0059] Instruments: GC-MS (Shimadzu QP2010 plus, Japan); microdroplet analyzer (Burkard, Burkard Manufacturing Co., Ltd., UK); digital constant-temperature magnetic stirrer (LC-DMS-H, Shanghai Lichen Bangsi Instrument Technology Co., Ltd.); circulating water multi-purpose vacuum pump (SHZ-D(Ⅲ), Tianjin Huaxin Instrument Factory); electric constant-temperature blast drying oven (DHG-9141, Shanghai Pudong Rongfeng Scientific Instrument Co., Ltd.); rotary evaporator (Hei-VAP(ML), Heidolph, Germany).

[0060] All experimental reagents not specifically described in the present invention are conventional reagents in the art, or are prepared using conventional methods in the art, are commercially available, and are of laboratory pure grade.

[0061] The first group of examples, application of Artemisia sphaerocephala in preventing and controlling agricultural pests

[0062] This group of embodiments provides the use of Artemisia sphaerocephala in preventing and controlling agricultural pests, wherein the agricultural pests are selected from western flower thrips, wolfberry psyllids or cotton aphids.

[0063] In some embodiments, the controlling refers to killing;

[0064] In other embodiments, the killing method is contact killing or fumigation killing.

[0065] In a further embodiment, Artemisia sphaerocarpa essential oil is used for insecticide.

[0066] Preferably, the Artemisia sphaerocarpa essential oil is an essential oil extracted from the aerial part of the plant;

[0067] Preferably, the active ingredients of the Artemisia sphaerocarpa essential oil include: biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cubediol, and α-pinene.

[0068] Second Group of Examples: Agents for Controlling Agricultural Pests of the Present Invention

[0069] All embodiments in this group have the following common features: the active ingredient of the agent for controlling agricultural pests includes: black artemisia essential oil; the agricultural pests are selected from western flower thrips, wolfberry psyllids or cotton aphids.

[0070] In a specific embodiment, the Artemisia sphaerocarpa essential oil is an essential oil extracted from the aerial part of the plant;

[0071] Preferably, the active ingredients of the Artemisia sphaerocarpa essential oil include: biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cubediol, and α-pinene.

[0072] In a specific embodiment, the dosage form of the agent for controlling agricultural pests is selected from the group consisting of: sprays, powders, tablets, capsules, and liquid preparations;

[0073] Preferably, the agent for controlling agricultural pests further comprises a pharmaceutically acceptable excipient.

[0074] In a specific embodiment, the pesticide for controlling agricultural pests includes but is not limited to: pesticides.

[0075] More specifically, the pesticide for controlling agricultural pests can be prepared into conventional dosage forms in the art, including but not limited to: dusts, wettable powders, drops, suspension concentrates, emulsifiable concentrates, sprays, aerosols, poison baits, mosquito coils, sticky traps, smoke agents, insecticidal paints, etc.

[0076] The third embodiment of the present invention is a method for preparing the biological control agent

[0077] This group of embodiments provides a pesticide for controlling agricultural pests. All embodiments in this group have the following common feature: Artemisia sphaerocarpa essential oil is used as the active ingredient of the pesticide for controlling agricultural pests.

[0078] In a specific embodiment, the aerial parts of the Artemisia sphaerocarpa plant are extracted by steam distillation and then extracted;

[0079] Preferably, the extraction temperature of the steam distillation method is 70-120°C, preferably 90°C; the extraction time is 4-12 h, preferably 6 h;

[0080] Preferably, the extractant is n-hexane, and the extraction time is 2-6 hours, preferably 3 hours;

[0081] Preferably, the n-hexane layer after extraction is dried and concentrated; preferably, the drying refers to removing moisture from the n-hexane layer with anhydrous sodium sulfate; and the concentration refers to removing n-hexane from the dried product by rotary evaporation using a rotary evaporator to obtain the black artemisia essential oil;

[0082] Preferably, the above-ground part of the Artemisia sphaerocephala plant is naturally dried in the shade and chopped before extraction by steam distillation;

[0083] Preferably, the steam distillation extraction method comprises: placing the air-dried and chopped above-ground parts of the Artemisia sphaerocephala plant in a round-bottom flask and extracting using an essential oil extractor;

[0084] Preferably, the method for preparing the agent for controlling agricultural pests further comprises: adding pharmaceutically acceptable excipients;

[0085] In a further embodiment, the agent for controlling agricultural pests further comprises excipients commonly accepted in the field of pesticides in the art, including but not limited to: emulsifiers, organic solvents, fillers, wetting agents, diluents, and the like.

[0086] In some embodiments, the method for preparing a pesticide for controlling agricultural pests further comprises: preparing a liquid or microcapsule formulation of the pesticide;

[0087] The preparation of the liquid preparation comprises: dissolving the black artemisia essential oil in a solvent; the solvent is preferably ethanol;

[0088] The preparation of the microcapsule comprises: using the inclusion complexation method to dropwise add a core material solution containing black artemisia essential oil into a wall material solution, stirring, standing, filtering, removing unreacted essential oil, and drying.

[0089] In a specific embodiment, the weight ratio of the core material solution to the wall material solution is 1:4-1:8;

[0090] Preferably, the core material solution is obtained by dissolving Artemisia sphaerocarpa essential oil in ethanol solution and stirring until completely dissolved;

[0091] Preferably, the wall material solution is obtained by dissolving β-cyclodextrin in deionized water and stirring until completely dissolved;

[0092] Preferably, the dissolution temperature is 30-60°C, preferably 50°C;

[0093] Preferably, the cyclodextrin mass concentration is 2.83% w / v;

[0094] Preferably, the stirring speed is 200-1000 r / min, preferably 500 r / min;

[0095] Preferably, the stirring time is 1-2 h;

[0096] Preferably, the standing is standing in a refrigerator at 4°C for 24 hours;

[0097] Preferably, the removal of unreacted essential oil is performed by washing with ethanol to remove the unreacted essential oil;

[0098] Preferably, the drying temperature is 30-60°C, preferably 40°C;

[0099] Preferably, the drying time is 3-6 h, preferably 4 h.

[0100] The fourth group of embodiments, the method for controlling agricultural pests of the present invention

[0101] This group of embodiments provides a method for controlling agricultural pests. All embodiments in this group have the following common features: the method for controlling agricultural pests uses Artemisia sphaerocephala to control agricultural pests; the agricultural pests are selected from western flower thrips, wolfberry psyllids, or cotton aphids.

[0102] Preferably, the black artemisia essential oil solution is used to control agricultural pests; the control of agricultural pests refers to killing insects;

[0103] In a specific embodiment, the insecticide method is selected from: contact killing or fumigation killing;

[0104] In some embodiments, the half-lethal concentration of the Artemisia sphaerocephala essential oil solution for killing cotton aphids is 5.87 mg / mL;

[0105] In some embodiments, the LC50 of the Artemisia sphaerocephala essential oil solution for killing western flower thrips is 1.57 mg / mL;

[0106] In some embodiments, the half-lethal concentration of the Artemisia sphaerocephala essential oil solution for killing Lycium barbarum psyllids is 0.81 mg / mL.

[0107] Experimental Example 1: Preparation of Artemisia sphaerocarpa essential oil and verification of insecticidal effect

[0108] 1.1 Preparation of Artemisia sphaerocarpa essential oil

[0109] The essential oil of 10 kg of the aerial parts of Artemisia sphaerocephala plants was naturally dried in the shade and obtained by steam distillation for 6 h. The essential oil was extracted with n-hexane and concentrated, then placed in dry glass bottles and stored in a refrigerator at 4°C. The oil yield (v / w) and its density were calculated.

[0110] 1.2 GC-MS analysis of Artemisia sphaerocarpa essential oil

[0111] GC conditions: GCMS-QP2010 plus (Shimadzu), chromatographic column DB-5MS (60 m´0.25 um´0.32 mm) (Agilent), source temperature 200 °C, interface temperature 300 °C, injection port temperature 320 °C, heating rate 60 °C to 10 °C / min, first at 300 °C for 20 min, followed by split injection with a ratio of 30:1, and constant linear velocity mode of 30 cm / s.

[0112] MS conditions: carrier gas He ion, mass scan range 29-650 m / z, injection volume 1 μL, the sample was filtered through a disposable syringe containing a 0.22 μm organic filter membrane filter, and the spectral library was searched against the NIST 2017 standard spectral library.

[0113] Artemisia sphaeroceae essential oil was extracted from Artemisia sphaeroceae by steam distillation for 6 hours. The calculated oil yield was 0.453%, and the oil density was 0.86 g / mL, calculated by weight-to-volume ratio. GC-MS analysis of the essential oil components was performed. The mass spectra were searched against the NIST 2017 mass spectral database and compared with standard spectra. Twenty-eight components were detected in the essential oil (Table 1), accounting for 99.35% of the total essential oil. The main component was biphenyl (15.62%). Other major components included bisabolol (10.11%), (-)-spartol (7.10%), and α-curcumene (6.06%).

[0114] Table 1. Components of Artemisia sphaerocarpa essential oil

[0115]

[0116]

[0117] The total percentage of all components identified in Table 1 above is less than 100% because there are still unknown components in the essential oil of Artemisia sphaerocarpa, which may be new compounds that cannot be identified using the existing GC-MS detection method.

[0118] 1.3 Determination of the insecticidal activity of Artemisia sphaerocephala essential oil against three pests: cotton aphid, wolfberry psyllid, and western flower thrips

[0119] Cotton aphid contact activity assay: Using the leaf disc method, ethanol was used as the solvent. The black artemisia essential oil prepared in step 1.1 was dissolved in the solvent to prepare five concentrations (2.14, 4.27, 8.55, 17.10, and 34.20 mg / mL). Leaf discs with a diameter of 12 mm were cut from cotton leaves and immersed in each concentration solution for 10 seconds. After drying naturally, the discs were placed in a twelve-well culture plate covered with an agar layer. The front of the leaf was completely attached to the bottom of the culture plate and in contact with the wall of the culture plate. Ten cotton aphid adults were placed in each well, and the culture plate was sealed with a rice paper strip. An aqueous solution containing equal amounts of anhydrous ethanol and DMSO was used as a blank control, and pyrethrum extract was used as a control. Each treatment was repeated 6 times, and the culture plate was placed in an incubator. The incubator conditions were: temperature (26±0.5)℃, relative humidity 50%, and light L:D of 16:8. The number of test insect deaths was observed and recorded after 24 hours. The LC was calculated using DPS software. 50 values and 95% confidence intervals.

[0120] Contact toxicity assay for Lycium barbarum psyllids: Five concentrations (0.24, 0.60, 1.52, 3.80, and 9.50 mg / mL) were prepared using a microdrip method. Test insects were anesthetized by freezing for 5–10 minutes. Ten Lycium barbarum psyllids were placed in a tube and the drug was dripped onto the pronotum. A control treatment was included, and the experiment was repeated six times. Treated Lycium barbarum psyllid adults were placed in a centrifuge tube lined with fresh Lycium barbarum buds and sealed with gauze. A control treatment containing no drug was also included. After treatment, the centrifuge tubes were placed in a plant incubator. Incubation conditions and data processing methods were as described above.

[0121] Fumigation activity assay for western flower thrips: Acetone was prepared at five concentrations (8.90, 17.80, 35.60, 71.20, and 142.40 mg / mL) using a fumigation method. Ten adult western flower thrips were inhaled into 20 mL vials, which were then covered with mesh and placed in a 250 mL wide-mouth bottle. A 1 cm × 6 cm filter paper strip was secured in the bottle cap. The filter paper strip, dripped with 20 μL of the solution, was allowed to air-dry for 10–20 s. After the solvent evaporated, the bottle cap was closed, leaving the filter paper strip suspended in the center of the wide-mouth bottle. The bottle cap was then covered with parafilm. Acetone solution served as a blank control, and neem oil served as a control. Six replicates were performed for each concentration. After treatment, the bottles were placed in an incubator set at (26 ± 0.5)°C, 50% relative humidity, and a light intensity of 16 h / / 8 h L / / D. After 24 hours, the number of dead insects in each treatment was investigated and recorded, and the data processing method was the same as above.

[0122] Indoor bioactivity test showed that the LC value of Artemisia sphaerocephala essential oil against cotton aphids was 50 The value is 5.87 mg / mL, which is higher than the control agent pyrethrum extract (3.54 mg / mL), and the toxicity is lower than the control agent pyrethrum extract; the LC 50 The value is 0.81 mg / mL, which is lower than the control agent pyrethrum extract (1.11 mg / mL) and higher in toxicity than the control agent pyrethrum extract; the LC 50 The value was 1.57 mg / mL, which was lower than that of the control agent pyrethrum extract (3.42 mg / mL) and higher in toxicity than that of the control agent neem oil.

[0123] Table 2. Insecticidal activity of Artemisia sphaerocephala essential oil against cotton aphids, wolfberry psyllids, and western flower thrips

[0124]

[0125] 1.4 Preparation of Artemisia sphaerocarpa essential oil microcapsules

[0126] Preparation by inclusion complexation: 8 g of β-cyclodextrin (2.83% w / v) was dissolved in deionized water at 50°C and stirred at 500 rpm until completely dissolved to obtain the wall material solution. 1.0 g of Artemisia sphaerocephala essential oil was dissolved in 25% ethanol and stirred until completely dissolved to obtain the core material solution. The core material solution was added dropwise to the wall material solution at a core-to-wall ratio of 1:4-1:8. Stirring was continued at 40-60°C for 1-2 hours, followed by storage in a refrigerator at 4°C. After 24 hours, the solution was filtered and rinsed with ethanol to remove unreacted essential oil. The solution was then placed in an oven at 40°C for 4 hours to obtain a white solid powder, which is the Artemisia sphaerocephala essential oil microcapsule. The prepared complex was placed in a round-bottom flask containing zeolite, added with an appropriate amount of distilled water, and connected to an essential oil analyzer. The mixture was boiled until the oil level in the analyzer no longer increased. The mixture was then removed from the oven and allowed to stand for a while. Open the piston and allow the oil layer to drop until its upper end is flush with the 0 line on the scale, and read the oil level.

[0127]

[0128]

[0129] Artemisia sphaerocarpa essential oil was prepared into microcapsules by inclusion complexation method with a core-wall ratio of 1:6, a preparation temperature of 60℃, and a stirring time of 120 min. The maximum encapsulation efficiency was 46% and the loading efficiency was 7.06%.

[0130] 1.5 Evaluation of the control effect of Artemisia sphaerocephala essential oil microcapsule preparation in the field

[0131] Experimental Design:

[0132] There are two test agents and one control agent (pyrethrin emulsion). The test agents are set at three concentrations (3, 6, 12 kg·hm -2 ), the control agent was treated with one concentration (6 kg·hm -2 ), spraying with clean water was used as the control. There were 4 treatments in total, and each treatment was repeated 4 times.

[0133] Survey time, frequency and method:

[0134] Before spraying, the baseline insect population on branches was surveyed. The remaining insect population was then surveyed six times on days 1, 3, 7, 14, and 21 after spraying. The specific survey method was as follows: Five branches were sampled from different directions within each plot, and the number of wolfberry pests within 30 cm of the top of each branch was recorded.

[0135] And calculate according to the following formula:

[0136]

[0137]

[0138] DPS statistical software was used to conduct variance analysis and significance analysis on the field control effect.

[0139] Field efficacy tests showed that the control effect of black artemisia essential oil microcapsules on wolfberry aphids was significantly lower than that of the control agent pyrethrum at the same dose after 1, 3 and 7 days of application; 14 days after application, the control effect of black artemisia essential oil microcapsules was significantly higher than that of the control agent at the same dose, and the insecticidal effect of the control agent pyrethrum was significantly reduced; 21 days after application, the control effect of black artemisia essential oil microcapsules at 6 kg·hm -2 The control effect of the wolfberry cotton aphid reached the maximum at the used dosage, which was 52.20%. The control effect was significantly higher than that of the control agent and the duration of effect reached more than 21 days.

[0140] Artemisia sphaerocarpa essential oil microcapsule preparation at 6 kg·hm -2 The control effect of the black artemisia essential oil microcapsules on wolfberry psyllids at the used doses was significantly different from that of the control agent pyrethrum; after 1 day, 7 days, 14 days, and 21 days of application, the control effect of the black artemisia essential oil microcapsules on wolfberry cotton aphids was significantly higher than that of the control agent pyrethrum at the same dose. 7 days after application, the black artemisia essential oil microcapsules achieved the maximum control effect of 58.41% on wolfberry cotton aphids; 14 days after application, there was no control effect on wolfberry cotton aphids; the control effect on wolfberry psyllids was significantly higher than that of the control agent, while the control agent pyrethrum had no insecticidal activity 14 days after application.

[0141] After 1, 3, and 7 days of application, the control effect of black artemisia essential oil microcapsules on thrips was significantly lower than that of the control agent pyrethrum at the same dose; after 14 and 21 days of application, the control effect of black artemisia essential oil microcapsules on western flower thrips was significantly higher than that of the control agent pyrethrum at the same dose. The control agent pyrethrum had no insecticidal effect, and the control effect improved with the increase of the dosage.

[0142] Table 3. Control effects of Artemisia sphaerocephala essential oil microcapsules on wolfberry aphids, wolfberry psyllids, and western flower thrips in the field

[0143]

[0144] Note: Data in the table are mean ± standard error. Different letters after the data in the same column indicate significant differences at the P < 0.05 level after one-way ANOVA test, while the same letters indicate no significant differences (P > 0.05).

[0145] Experimental Example 2: Killing experiment of Artemisia selengensis essential oil on Mythimum selengensis

[0146] In Experimental Example 2, the leaf disc method was used. Five concentrations of Artemisia sphaerocephala essential oil were prepared (12.5, 25, 50, 100, and 200 mg / mL). Adult Mythimna separata was used as the test insect. Other operations were the same as those in Section 1.3 "Determination of contact activity against cotton aphids" of Experimental Example 1.

[0147] Table 4. Determination of the killing activity of Artemisia sphaerocephala essential oil against armyworms

[0148]

Claims

1. Application of Artemisia sphaerocephala in preventing and controlling agricultural pests, wherein the agricultural pests are selected from western flower thrips, wolfberry psyllids or cotton aphids; using Artemisia sphaerocephala essential oil to prevent and control agricultural pests; the Artemisia sphaerocephala essential oil is an essential oil extracted from the aerial parts of the plant; the aerial parts of the Artemisia sphaerocephala plant are extracted by steam distillation and then extracted; the extractant is n-hexane.

2. The use of Artemisia sphaerocephala for preventing and controlling agricultural pests according to claim 1, characterized in that: The active ingredients of the black artemisia essential oil include biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cube alcohol, and α-pinene.

3. A medicament for controlling agricultural pests, characterized in that: Its active ingredients include: black wormwood essential oil; the agricultural pests are selected from western flower thrips, wolfberry psyllids, and cotton aphids; black wormwood essential oil is used to control agricultural pests; the black wormwood essential oil is extracted from the aboveground part of the plant; the aboveground part of the black wormwood plant is extracted by steam distillation and then extracted; the extractant is n-hexane.

4. The agent for controlling agricultural pests according to claim 3, characterized in that: The active ingredients of the black artemisia essential oil include biphenyl, bisabolol, (-)-spartol, α-curcumene, nerolidol, β-pinene, cube alcohol, and α-pinene.

5. The agent for controlling agricultural pests according to claim 3 or 4, characterized in that: The pesticide for preventing and controlling agricultural pests is selected from the group consisting of sprays, powders, tablets, microcapsules, and liquid preparations.

6. The agent for controlling agricultural pests according to claim 3 or 4, characterized in that: The agent for controlling agricultural pests further comprises pharmaceutically acceptable adjuvants.

7. A method for preparing a medicament for controlling agricultural pests, characterized in that: include: Using black artemisia essential oil as the active ingredient of a medicament for controlling agricultural pests; the agricultural pests are selected from western flower thrips, wolfberry psyllids or cotton aphids; Black Artemisia sphaeroceae essential oil is used to control agricultural pests; the black Artemisia sphaeroceae essential oil is essential oil extracted from the above-ground part of the plant; the above-ground part of the black Artemisia sphaeroceae plant is extracted by steam distillation and then extracted; the extraction agent is n-hexane.

8. The method for preparing a medicament for controlling agricultural pests according to claim 7, characterized in that: The extraction temperature of the steam distillation method is 70-120° C.; the extraction time is 4-12 hours.

9. The method for preparing a medicament for controlling agricultural pests according to claim 8, characterized in that: The extraction temperature of the steam distillation method is 90° C. and the extraction time is 6 hours.

10. The method for preparing a medicament for controlling agricultural pests according to claim 7, characterized in that: The extraction time is 2-6h.

11. The method for preparing a medicament for controlling agricultural pests according to claim 10, characterized in that: The extraction time is 3h.

12. The method for preparing a medicament for controlling agricultural pests according to claim 7, characterized in that: The extracted n-hexane layer was dried and concentrated.

13. The method for preparing a medicament for controlling agricultural pests according to claim 12, characterized in that: The drying refers to removing moisture from the n-hexane layer with anhydrous sodium sulfate; The concentration refers to removing n-hexane from the dried product by rotary evaporation using a rotary evaporator to obtain the black artemisia essential oil.

14. The method for preparing a pesticide for controlling agricultural pests according to claim 7, characterized in that: Before extraction by steam distillation, the above-ground part of the Artemisia sphaerocephala plant is naturally dried in the shade and cut into pieces.

15. The method for preparing a pesticide for controlling agricultural pests according to claim 14, characterized in that: The steam distillation extraction method comprises placing the shade-dried and chopped above-ground parts of the Artemisia sphaerocephala plant in a round-bottom flask and extracting the essential oil using an essential oil extractor.

16. A method for preparing a pesticide for controlling agricultural pests according to any one of claims 7 to 14, characterized in that: Also includes: Add pharmaceutically acceptable excipients.

17. A method for preparing a pesticide for controlling agricultural pests according to any one of claims 7 to 14, characterized in that: The agent for controlling agricultural pests is selected from a liquid agent and a microcapsule agent; the liquid agent is a black artemisia essential oil solution obtained by dissolving black artemisia essential oil in ethanol; the microcapsule agent adopts an inclusion complexation method to add a core material solution containing black artemisia essential oil dropwise to a wall material solution, stir, and then stand, and then filter to remove unreacted essential oil and then dry; The weight ratio of the core material solution to the wall material solution is 1:4-1:8; The core material solution is obtained by dissolving black artemisia essential oil in ethanol solution and stirring until it is completely dissolved; The wall material solution is obtained by dissolving β-cyclodextrin in deionized water and stirring until completely dissolved; The cyclodextrin mass concentration is 2.83% w / v; The standing is standing in a 4°C refrigerator for 24 hours; The unreacted essential oil is removed by washing with ethanol; The drying temperature is 30-60°C; The drying time is 3-6 h.

18. The method for preparing a pesticide for controlling agricultural pests according to claim 17, wherein: The drying temperature is 40° C. and the drying time is 4 h.

19. A method for controlling agricultural pests, characterized in that: Black Artemisia ordosica is used to control agricultural pests; the agricultural pests are selected from western flower thrips, wolfberry psyllids, and cotton aphids; black Artemisia ordosica essential oil solution is used for pest control; the black Artemisia ordosica essential oil is extracted from the aboveground part of the plant; the aboveground part of the black Artemisia ordosica plant is extracted by steam distillation and then extracted; the extractant is n-hexane.

20. The method for controlling agricultural pests according to claim 19, characterized in that: The insecticide method is selected from: contact killing or fumigation killing.

21. The method for controlling agricultural pests according to claim 20, wherein: The half-lethal concentration of the Artemisia sphaerocephala essential oil solution for killing cotton aphids is 5.87 mg / mL.

22. The method for controlling agricultural pests according to claim 20, wherein: The half lethal concentration of the Artemisia sphaerocephala essential oil solution for killing western flower thrips is 1.57 mg / mL.

23. The method for controlling agricultural pests according to claim 20, wherein: The half lethal concentration of the black artemisia essential oil solution for killing wolfberry psyllids is 0.81 mg / mL.

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