Method for preventing and treating diaphorina citri

Abstract

The invention provides a biological source compound for preventing and treating diaphorina citri and a method thereof, the biological source compound for preventing and treating diaphorina citri comprises the component of Asperparaline A. The Asperparaline A is biosynthesized through Aspergillus Fiji YS-1, the Asperparaline A is efficiently purified and prepared into the compound with the concentration of 95 ppm, the compound is used for preventing and treating diaphorina citri adults, the prevention and treatment effect is extremely good, and the compound has a good application prospect in prevention and treatment of diaphorina citri adults. And the composition also has a relatively good prevention and treatment effect on diaphorina citri nymphs. The biosynthesized compound is applied to prevention and treatment of diaphorina citri for the first time, the prevention and treatment effect is ideal, and due to the fact that the compound is a biological source compound, the compound is environmentally friendly and green and has wide application prospects.

Description

[Technical field]

[0001] The invention belongs to the technical field of green control of fruit tree pests, and relates to a method for controlling citrus psyllids, a method for controlling citrus psyllids by utilizing biogenic compounds, a method for biosynthesizing Asperparaline A by utilizing Aspergillus Fijiensis YS-1 and using the Asperparaline A for controlling citrus psyllids, and an application method for creating a new control agent for citrus psyllids. [Background technology]

[0002] Citrus Huanglongbing (HLB) is an infectious and devastating disease in citrus production, and there is currently no complete cure. Therefore, the prevention and control of citrus psyllids has become the focus of preventing the rampant occurrence and spread of citrus Huanglongbing. The abuse and repeated use of pesticides have caused citrus psyllids to develop a high degree of resistance to many pesticides, and there is an urgent need to develop new biogenic compounds for the prevention and control of citrus psyllids.

[0003] Asperparaline A is an isopentenyl indole alkaloid containing a [2,2,2]-cyclodiazooctane ring system. It was first reported as a metabolite produced by Aspergillus japonicus JV-23 and has anthelmintic activity against the third-instar larvae of parasitic nematodes in the digestive tract of mammals. Studies have further found that the substance exhibits insecticidal activity against agricultural pests such as brown planthoppers (International Patent Publication No. 2010 / 071218). In addition, studies have also reported that the substance exhibits insecticidal activity against animal parasitic pests such as hard ticks and fleas, and is safe for mammals (International Patent Publication No. 2010 / 071219).

[0004] In summary, Asperparaline A has the potential to be used as an insecticide and can be developed as a new microbial pesticide. [Summary of the invention]

[0005] In view of the fact that there are few pesticides registered for the control of citrus psyllids in the prior art, and the only microbial pesticide is avermectin, and the citrus psyllids have increased resistance to existing agents, resulting in the gradual weakening of the effects of most agents, it is urgent to develop new microbial pesticides. The purpose of the present invention is to provide a method for controlling citrus psyllids, a method for efficiently controlling citrus psyllids using a microbial compound Asperparaline A, and a method for efficiently controlling Asian citrus psyllids.

[0006] During a field investigation in Yangshuo County, Guilin City, the inventor discovered dead adult citrus psyllids, and isolated multiple fungi from the corpses. One of them was identified as Aspergillus fijiensis, named AspergillusfijiensisYS-1, and was preserved in the Guangdong Provincial Microbiological Culture Collection Center on July 7, 2020, with the preservation number GDMCCNo.61081.

[0007] The technical solution of the present invention is as follows:

[0008] A method for controlling citrus psyllids comprises the following steps:

[0009] S1. Fermentation culture of biocontrol fungi: Fermentation culture of Aspergillus fijiensis YS-1 strain to obtain fungal spores;

[0010] The Aspergillus fijiensis YS-1 strain is a fungus isolated from the dead bodies of adult citrus psyllids in the field in Yangshuo County, Guilin City. One of the strains was identified as Aspergillus fijiensis, named Aspergillus fijiensis YS-1, and was preserved in the Guangdong Provincial Microbiological Culture Collection Center on July 7, 2020. The preservation address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, and the preservation number is GDMCC No. 61081;

[0011] S2. Extracting Asperparaline A: extracting Aspergillus fijiensis YS-1 spores with an alcohol solution and concentrating to obtain an extract; dispersing and dissolving the extract with water, performing column chromatography gradient elution, collecting, combining and concentrating specific eluates to obtain a concentrated eluate; standing the concentrated eluate, recrystallizing, filtering the mother liquor, and obtaining crystals that are Asperparaline A;

[0012] S3. Preparation of control agent: dilute Asperparaline A with water to 95 ppm;

[0013] S4. Control of citrus psyllids: During each citrus shoot growth period, when the new shoots grow to 0.5-1.0 cm, use a sprayer to spray the entire citrus tree with Asperparaline A diluted to 95 ppm. Spray each tree until both sides of the leaves are completely soaked and dripping.

[0014] In step S1, the fermentation culture is carried out indoors using potato dextrose agar medium (PDA) or other solid culture medium, the culture time is 3-15 days, and the culture temperature is 20-38°C.

[0015] In step S2, the alcohol solution is a methanol or ethanol solution with a volume fraction of 30-95% and a solid-liquid ratio (g:mL) of 1:10-1:40.

[0016] In step S2, the extraction method is ultrasonic-assisted extraction, and the ultrasonic-assisted extraction conditions are as follows: 100-300 g of YS-1 spore raw material is taken into a conical flask, 1000-6000 mL of a methanol solution with a volume fraction of 30-95% is added thereto, and ultrasonic extraction is performed at 30-80° C. for 30-90 min, with a power of 200-800 W and a frequency of 20-80 KHz. The extraction is repeated 2-5 times, and then the solvent is removed by negative pressure distillation to obtain an extract.

[0017] In step S2, the extract is dispersed and dissolved with water by weighing the mass (g) of the extract, adding 5-20 times volume (mL) of pure water to dissolve it, and waiting for wet column loading.

[0018] In step S2, column chromatography gradient elution is performed, wherein the resin for column chromatography is selected from D101 macroporous resin.

[0019] In step S2, the column chromatography gradient elution is performed, and specific eluents are collected, combined and concentrated. Specifically, D101 macroporous resin is used as the stationary phase, and the elution conditions are: elution is performed with ethanol solutions with volume fractions of 15%, 25%, 30%, and 40% in sequence, each gradient elution volume is 1.0-10.0BV, the flow rate is controlled to be 0.5-5.0BV / h, and the eluents of 30% and 40% ethanol are collected, concentrated and combined.

[0020] In step S2, the purification method includes crystallization, precipitation, membrane separation, preparative liquid chromatography, silica gel column chromatography or ODS column chromatography.

[0021] In step S2, the recrystallization is replaced by waiting for natural precipitation crystallization process.

[0022] More specifically,

[0023] The method for controlling citrus psyllids of the present invention is a method for using Aspergillus fijiensis YS-1 to biosynthesize Asperparaline A, and preparing it into a medicament after purification for controlling citrus psyllids, comprising the following steps:

[0024] S1: Acquisition of Aspergillus Fijiensis YS-1:

[0025] Dead citrus psyllid adults were found in the fields of Yangshuo County, Guilin City. Several fungi were isolated from the dead insects, one of which was identified as Aspergillus fijiensis and named Aspergillus fijiensis YS-1.

[0026] S2: Cultivation and spore collection of Aspergillus Fijiensis YS-1:

[0027] The Aspergillus fijiensis YS-1 spores from the citrus psyllid were inoculated on 2000 PDA culture media, and after continuous cultivation in the room for 7 days, the Aspergillus fijiensis YS-1 spores were shaken off into a conical flask to obtain spores with strong infectivity.

[0028] S3: Extraction of secondary metabolites from Aspergillus Fijiensis YS-1:

[0029] Weigh 150g of the YS-1 spore material obtained in the previous step, add 4L of 85% methanol by volume, divide into 4 portions in a conical flask, perform ultrasonic extraction at 50°C for 60min, repeat the extraction three times, and then remove the solvent by negative pressure distillation. After combining, 50.0g of extract is obtained, and 500ml of pure water is added to dissolve and disperse to obtain a concentrated solution;

[0030] S4: Purification and extraction of Asperparaline A:

[0031] S4-1: Preparation of macroporous resin chromatography column:

[0032] Select 5.0L of D101 macroporous resin, add 95% ethanol to it, let it stand for 24 hours, then add it to an empty chromatography column, wait for it to settle naturally, then use ethanol to pass through the resin layer at a flow rate of 2.0BV / h, wash until the effluent is free of white turbidity, then replace the ethanol with distilled water, and rinse until the effluent has no alcohol taste; then use 2.0BV 5% (v / v) HCl solution to pass through the resin layer at a flow rate of 4.0-6.0BV / h, and soak for 2-4 hours, then use water to wash at the same flow rate until the pH of the effluent is neutral, then use 2.0BV of 2.0% (w / v) NaOH solution to pass through the resin layer at a flow rate of 4.0-6.0BV / h, and soak for 2-4 hours, then wash with water at the same flow rate until the pH of the effluent is neutral;

[0033] S4-2: Separation and purification:

[0034] The concentrated solution obtained from S3 was slowly drained into the macroporous resin chromatography column with a glass rod. After the sample was added, it was allowed to stand for 30 minutes to allow the sample to be fully adsorbed on the macroporous resin; gradient elution was performed with 15%, 25%, 30%, and 40% ethanol solutions by volume fraction in sequence, and finally the chromatography column was rinsed with 95% ethanol solution. The volume of each gradient elution was 1.0 BV, and the flow rate was controlled to be 2.0 BV / h. The eluates of 30% and 40% ethanol were concentrated by rotary evaporator under reduced pressure and then combined, and allowed to stand overnight at 4°C to wait for the crystals to precipitate automatically. The chromatography column rinsed with 95% ethanol solution can be reused.

[0035] S5: Confirmation of extracted Asperparaline A:

[0036] Single crystals were selected and analyzed for purity and structure by HPLC, X-ray single crystal diffraction, nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HR-MS), and compared with literature and Pubchem database to confirm that the extracted and purified compound was Asperparaline A.

[0037] S5: Preparation of citrus psyllid control agent:

[0038] Asperparaline A was diluted with water to 95 ppm.

[0039] S6: Control of citrus psyllids:

[0040] During each citrus shoot growth period, when the new shoots grow to 0.5-1.0 cm, Asperparaline A diluted to 95 ppm is sprayed on the entire citrus tree using a sprayer. Each tree is sprayed until both sides of the leaves are completely soaked and dripping.

[0041] Compared with the prior art, the present invention has the following advantages:

[0042] 1. A method for controlling citrus psyllids described in the present invention is the application of Asperparaline A in controlling citrus psyllids, which has strong toxicity and excellent effect: since the present invention is a new agent, citrus psyllids have no resistance to it, so that the insecticidal toxicity of the compound is strong. Indoor biological test results show that the lethal effect of Asperparaline A on citrus psyllids is better than that of common pesticides such as imidacloprid.

[0043] 2. The Asperparaline A of the present invention is environmentally friendly: Although the chemical control methods used in the prior art have played a certain role in preventing and controlling the damage of citrus psyllids, they also cause pollution, damage and residues to humans, livestock and farmland ecosystems; the present invention is a microbial source pesticide, which is derived from biosynthesis and contains only four common chemical elements: C, H, O and N. Compared with synthetic organophosphorus pesticides, it is easier to degrade in the environment and is more environmentally friendly.

[0044] 3. The method for controlling citrus psyllids of the present invention has stable effects and is easy to store: plant-derived pesticides have the disadvantage of being easily ineffective, such as matrine, which easily loses its efficacy in adverse environments and must be supplemented with additives to stabilize its chemical properties, while Asperparaline A has a relatively stable chemical structure.

[0045] 4. The method for controlling citrus psyllids of the present invention has no odor: artificially synthesized insecticides (especially some organic phosphorus and sulfur-containing pesticides) often have a foul odor and have an impact on the environment. Asperparaline A has no obvious odor and has potential application prospects in urban landscaping pest control.

[0046] 5. The method for controlling citrus psyllids of the present invention has good water solubility: Asperparaline A has high polarity and is easily soluble in methanol solution. It also has good solubility in water without adding organic solvents, co-solvents and surfactants, which is conducive to the development of aqueous pesticides, reducing formulation costs and reducing pressure on the environment.

[0047] 6. The method for controlling citrus psyllids described in the present invention can provide a novel lead compound for the artificial synthesis of pesticides: Asperparaline A is a special isopentenyl indole alkaloid containing a [2.2.2]-cyclodiazooctane ring system, and its molecular structure is different from that of existing neonicotinoids, organophosphorus, and organochlorine insecticides. It can provide a lead compound for subsequent artificial synthesis and further structural modification in order to create new pesticides.

Brief Description of the Drawings

[0048] Figure 1 The growth morphology of Aspergillus fijiensis YS-1 on PDA medium in the embodiment of the present invention;

[0049] Figure 2 The molecular structure diagram of Asperparaline A in the embodiment of the present invention;

[0050] Figure 3This is a graph showing the change in mortality of citrus psyllid adults over time after treatment with different concentrations of Asperparaline A in an example of the present invention. [Specific implementation method]

[0051] The specific implementation modes of the present invention are further described below in conjunction with examples.

[0052] Example:

[0053] A method for controlling citrus psyllids comprises the following steps:

[0054] 1. Brief description:

[0055] This study provides a new biogenic compound Asperparaline A for controlling citrus psyllids and a method thereof. The method comprises the steps of biosynthesizing Asperparaline A from Aspergillus fijiensis YS-1, preparing an agent for controlling citrus psyllids, and applying it in the field.

[0056] 2. Obtaining Aspergillus Fijiensis YS-1:

[0057] Dead citrus psyllid adults were found in the fields of Yangshuo County, Guilin City. Several fungi were isolated from the dead insects. One of them was identified as Aspergillus fijiensis and named Aspergillus fijiensis YS-1 (see Figure 1 ).

[0058] 3. Cultivation and spore collection of Aspergillus Fijiensis YS-1:

[0059] The Aspergillus fijiensis YS-1 spores from the citrus psyllid were inoculated onto 2000 PDA culture media respectively. After continuous cultivation indoors for 7 days, the Aspergillus fijiensis spores were shaken off into a conical flask to obtain spores with strong infectivity.

[0060] 4. Extraction of secondary metabolites of Aspergillus Fiji YS-1:

[0061] Weigh 150g of the YS-1 spore raw material obtained in the previous step, add 4L of 85% methanol by volume, divide it into 4 parts in a conical flask, and ultrasonically extract it at 50°C for 60min. Repeat the extraction three times, and then remove the solvent by negative pressure distillation. After combining, 50.0g of extract is obtained, and 500ml of pure water is added to dissolve and disperse it to obtain a concentrated solution.

[0062] 5. Purification and extraction of Asperparaline A:

[0063] 5.1 Macroporous resin pretreatment:

[0064] Prepare about 5.0L of newly purchased D101 macroporous resin, add 95% ethanol to it, let it stand for 24 hours, and then add it to an empty chromatography column; after it is naturally precipitated, use ethanol to pass through the resin layer at a flow rate of 2.0BV / h, wash until the effluent is free of white turbidity, then replace the ethanol with distilled water, and rinse until the effluent has no alcohol taste; then use 2.0BV of 5% HCl solution to pass through the resin layer at a flow rate of 4.0-6.0BV / h, and soak for 2-4 hours, and then use water to wash at the same flow rate until the pH of the effluent is neutral; then use 2.0BV of 2.0% NaOH solution to pass through the resin layer at a flow rate of 4.0-6.0BV / h, and soak for 2-4 hours, and then wash with water at the same flow rate until the pH of the effluent is neutral;

[0065] 5.2 Separation and purification

[0066] The concentrated solution obtained in step 4 is slowly drained into the macroporous resin chromatography column with a glass rod. After the sample is added, it is allowed to stand for 30 minutes to allow the sample to be fully adsorbed on the macroporous resin; gradient elution is performed with ethanol solutions with volume fractions of 15%, 25%, 30%, 40%, and finally the chromatography column is rinsed with 95% ethanol solution. The volume of each gradient elution is 1.0BV, and the flow rate is controlled to be 2.0BV / h. The eluate of 30% and 40% ethanol solution is collected by equal flow fraction method (1L), and concentrated by rotary evaporator under reduced pressure. The concentrated solution is combined and allowed to stand overnight at 4°C to wait for the crystals to precipitate automatically. The chromatography column rinsed with 95% ethanol solution can be reused.

[0067] 6. Purity test and structural identification of Asperparaline A extract:

[0068] 6.1 HPLC Purity Test of Asperparaline A:

[0069] Instrument model: Shimadzu LC-20A;

[0070] Chromatographic column: inertsil ods-3 (4.6 mm id × 250 mm, 5 μm);

[0071] Mobile phase: water (phase A), methanol (phase B);

[0072] Gradient program: 0-25 min, B%: 5-100%;

[0073] Flow rate: 1 mL min -1 ;

[0074] Column temperature: 35°C;

[0075] Injection volume: 5.0 μL;

[0076] Wavelength: 206nm.

[0077] 6.2 Asperparaline A structure identification:

[0078] The extracted and purified Asperparaline A was analyzed by X-ray single crystal diffraction, nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HR-MS), and compared with the literature and Pubchem database to clarify the structure of Asperparaline A. Figure 2 shown.

[0079] 7. Toxicity determination of Asperparaline A against citrus psyllids:

[0080] 7.1 Toxicity of Asperparaline A to Citrus Psyllid Adults and Comparison with Other Insecticides:

[0081] 7.1.1 Test method:

[0082] (1) Pick out citrus psyllid adults of similar age and freeze them in a -20°C refrigerator for 1-2 minutes (short-term paralysis with CO2), and use a brush to pick out 30 of them and place them in a culture dish;

[0083] (2) Prepare 5 levels of the drug with clean water, with the control group being clean water;

[0084] (3) Place the culture dish device containing the test insects on the stage of the spray tower, take 1 mL of spore suspension and add it into the spray tower for spraying. Treat 90 test insects for each concentration, and the spraying time is 1 min.

[0085] (4) The treated test insects were placed at room temperature for 30 min, and then the psyllids were transferred to a disposable cup containing fresh citrus leaves containing psyllids;

[0086] (5) The test insects were raised in an artificial climate chamber at a temperature of 25±1°C, a relative humidity of 75%-85%, and a photoperiod of 16:8h (light:dark). The survival rate was counted every 12h until 72h.

[0087] 7.1.2 Test results:

[0088] Asperparaline A was more toxic to adults of citrus psyllid than imidacloprid, avermectin, sulfoxaflor and flupyralidone (see Table 1).

[0089] Table 1 Toxicity of Asperparaline A and other pesticides to citrus psyllid adults

[0090]

[0091] When df = 3, the χ2 value when P = 0.05 was 7.81. The actual χ2 values ​​of imidacloprid, thiamethoxam, avermectin, dinotefuran, sulfoxaflor, and flupyralidone were all smaller than the theoretical χ2, indicating that the toxicity regression equations were consistent with the actual situation.

[0092] The cumulative mortality of citrus psyllid adults after treatment with different concentrations of Asperparaline A was shown in Figure 3 The results showed that the cumulative mortality of citrus psyllid adults treated with Asperparaline A increased with the increase of treatment time (24, 48 and 72 hours). The corrected mortality rate after 72 hours of treatment with the highest concentration (50 mg / L) of Asperparaline A was 84.68%.

[0093] 7.1.3 Conclusion:

[0094] Asperparaline A, which is biosynthesized and purified by Aspergillus fijiensis YS-1, has a good control effect on adult citrus psyllids. It is superior to the commonly used pesticides for killing citrus psyllids in the field, such as imidacloprid, avermectin, sulfoxaflor and flupyradone, and therefore has great application potential.

[0095] 7.2 Toxicity of Asperparaline A to Citrus Psyllid Nymphs and Comparison with Other Insecticides:

[0096] 7.2.1 Test method:

[0097] (1) Use a brush to pick up 20 3-year-old citrus psyllid nymphs and place them in a culture dish;

[0098] (2) Prepare 5 levels of the drug with clean water;

[0099] (3) Place the petri dish device containing the test insects on the stage of the spray tower, take 1 mL of spore suspension and add it to the spray tower for spraying. Treat 60 test insects for each concentration, and the spraying time is 1 min;

[0100] (4) The treated test insects were placed at room temperature for 30 min, and then the citrus psyllid nymphs were transferred to a disposable cup containing fresh citrus leaves containing psyllids;

[0101] (5) The test insects were raised in an artificial climate chamber at a temperature of 25±1°C, a relative humidity of 75%-85%, and a photoperiod of 16:8h (light:dark). The survival rate was counted every 12h until 48h.

[0102] 7.2.2 Test results:

[0103] Asperparaline A was more toxic to citrus psyllid nymphs than dinotefuran and avermectin (see Table 2). 48 hours after treatment with 20 mg / L Asperparaline A, the corrected mortality of citrus psyllid nymphs was 77.92%.

[0104] Table 2 Toxicity of Asperparaline A and other agents to citrus psyllid nymphs

[0105]

[0106] 7.2.3 Conclusion:

[0107] Asperparaline A, which is biosynthesized and purified by Aspergillus fijiensis YS-1, also has a good control effect on citrus psyllid nymphs.

[0108] 7.3 Asperparaline A for controlling citrus psyllids in the field:

[0109] 7.3.1 Test method:

[0110] During the spring shoot period of citrus, when the new shoots grow to 0.5-1.0 cm, Asperparaline A diluted with water to 95 ppm was sprayed on the whole citrus tree with a sprayer. Each tree was sprayed until both sides of the leaves were completely soaked and dripping. In the test orchard, 9 points (including the plants on the edge of the orchard) were surveyed using the "Z" shaped sampling method, with 2 plants at each sampling point, and 1 new shoot of each plant was surveyed in each of the four directions of "east, south, west and north". The number of adult citrus psyllids was investigated.

[0111] 7.3.2 Test results:

[0112] Three days after spraying, through investigation in the experimental orchard, the reduction rate of citrus psyllid adults reached 97.2%, and the effect of field control of citrus psyllid was very good.

[0113] The above description is only a preferred embodiment of the present invention. It should be pointed out that it is possible for a person skilled in the art to make several improvements and changes without departing from the creative concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. A method for controlling citrus psyllids, characterized in that: The following steps are involved: S1. Fermentation culture of biocontrol fungi: Fermentation culture of Aspergillus fijiensis YS-1 strain to obtain fungal spores; The Aspergillus fijiensis YS-1 strain is a fungus isolated from the dead bodies of adult citrus psyllids in the field in Yangshuo County, Guilin City. One of the strains was identified as Aspergillus fijiensis, named Aspergillus fijiensis YS-1, and was preserved in the Guangdong Provincial Microbiological Culture Collection Center on July 7, 2020. The preservation address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, and the preservation number is GDMCC No. 61081; S2. Extracting Asperparaline A: extracting Aspergillus fijiensis YS-1 spores with an alcohol solution and concentrating to obtain an extract; dispersing and dissolving the extract with water, performing column chromatography gradient elution, collecting, combining and concentrating specific eluates to obtain a concentrated eluate; standing the concentrated eluate, recrystallizing, filtering the mother liquor, and obtaining crystals, which are Asperparaline A; S3. Preparation of control agent: dilute Asperparaline A with water to 95 ppm; S4. Control of citrus psyllids: During each citrus shoot growth period, when the new shoots grow to 0.5-1.0 cm, use a sprayer to spray the entire citrus tree with Asperparaline A diluted to 95 ppm. Spray each tree until both sides of the leaves are completely soaked and dripping.

2. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S1, the fermentation culture is carried out indoors using potato dextrose agar medium, the culture time is 3-15 days, and the culture temperature is 20-38°C.

3. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the alcohol solution is a methanol or ethanol solution with a volume fraction of 30-95%, and a solid-liquid ratio g:mL of 1:10-1:

40.

4. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the extraction method is ultrasonic-assisted extraction, and the ultrasonic-assisted extraction conditions are as follows: 100-300 g of YS-1 spore raw material is taken into a conical flask, 1000-6000 mL of a methanol solution with a volume fraction of 30-95% is added thereto, and ultrasonic extraction is performed at 30-80° C. for 30-90 min, with a power of 200-800 W and a frequency of 20-80 KHz. The extraction is repeated 2-5 times, and then the solvent is removed by negative pressure distillation to obtain an extract.

5. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the extract is dispersed and dissolved with water by weighing the mass of the extract in g, adding 5-20 times volume mL of pure water to dissolve it, and waiting for wet column loading.

6. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, column chromatography gradient elution is performed, wherein the resin for column chromatography is selected from D101 macroporous resin.

7. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the column chromatography gradient elution is performed, and specific eluents are collected, combined and concentrated. Specifically, D101 macroporous resin is used as the stationary phase, and the elution conditions are: elution is performed with ethanol solutions with volume fractions of 15%, 25%, 30%, and 40% in sequence, each gradient elution volume is 1.0-10.0BV, the flow rate is controlled to be 0.5-5.0BV / h, and the eluents of 30% and 40% ethanol are collected, concentrated and combined.

8. The method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the purification method includes crystallization, precipitation, membrane separation, preparative liquid chromatography, silica gel column chromatography or ODS column chromatography.

9. A method for controlling citrus psyllid according to claim 1, characterized in that: In step S2, the recrystallization is replaced by waiting for natural precipitation crystallization process.

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