A kind of wolfberry psyllid sex pheromone and its application

Abstract

The present invention discloses a sex pheromone for wolfberry psyllids and its application. The sex pheromone for wolfberry psyllids comprises 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, toluene, 2-hexanone, and / or 1-methylcyclopentanol. The present invention also provides a wolfberry psyllid sex attractant comprising 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, toluene, 2-hexanone, and / or 1-methylcyclopentanol as active ingredients, and further provides a wolfberry psyllid sex attractant core comprising a carrier and the wolfberry psyllid sex attractant loaded on the carrier. The method for monitoring and controlling wolfberry psyllids provided by the present invention has the advantages of being easily available raw materials for the discovered sex pheromone active ingredients, low cost, and a significant attractant effect. The method also has the advantages of being simple in technology, easy to operate, easy to commercialize and promote, and being environmentally friendly and sensitive. In addition, the method does not cause pesticide residues, can reduce the amount of chemical pesticides required for controlling wolfberry psyllids, and is conducive to alleviating the problem of high pesticide residues in wolfberries.

Description

Technical Field

[0001] The invention relates to a sex pheromone of wolfberry psyllid and application thereof, and belongs to the technical field of green pest prevention and control / pest monitoring and trapping. Background Art

[0002] The goji berry psyllid, Bactericera gobica, belongs to the family Psyllidae in the order Hemiptera. Its feeding can weaken plant growth and severely reduce branching in the current and following years. During outbreaks, the prevalence can reach over 80%, often leading to reduced goji berry yield and quality, and in severe cases, even complete crop failure (Liu Sai et al., 2020; Li Jianling et al., 2021). Goji berry psyllids reproduce sexually, making their control difficult due to their high reproductive capacity and high population density. Field surveys have found that a single goji berry leaf can contain 20 to 50 eggs, with four to five generations occurring annually. Therefore, the damage caused by goji berry psyllids has become a critical issue that urgently needs to be addressed in goji berry production. Currently, chemical pesticides are the primary control method for goji berry psyllids. Goji berry psyllids are characterized by their high reproductive capacity and significant generational overlap. Often, before the previous generation of adults is controlled, the next generation of nymphs has already emerged, making pest control extremely challenging. Furthermore, due to the specialized feeding habits of the wolfberry psyllid, which is highly resistant to pesticides, the indiscriminate and overuse of chemical pesticides has led to a rapid development of resistance. Consequently, current chemical pesticide control of the wolfberry psyllid is often ineffective. Furthermore, the infestation period of the wolfberry psyllid overlaps significantly with the wolfberry harvest season. The extensive use of chemical pesticides during this period to control the wolfberry psyllid leads directly to excessive pesticide residues in the wolfberries, seriously impacting their quality. Therefore, to reduce the use of chemical pesticides and address the problem of high pesticide residues in wolfberries, there is an urgent need to research and develop green control methods for the wolfberry psyllid.

[0003] Mating is a prerequisite for psyllid egg-laying and reproduction. Successful mating in insects requires courtship and copulation, processes often regulated by pheromones. The use of pheromones for pest control has been a new biological pest control technology developed since the 1960s (Karlson and Butenandt, 1959). Commercial products have now been developed and widely used in pest control (Fassotte et al., 2016; Mann et al., 2013). Compared with chemical pesticides, pheromone control offers advantages such as environmental friendliness, high efficiency, non-toxicity, high sensitivity, and resistance to pesticide resistance. It has become one of the preferred methods for pest monitoring and ecological control in modern agricultural production (Benelli et al., 2019; Liu et al., 2022; Knight et al., 2019; Zisopoulou et al., 2020). Researchers at home and abroad have conducted extensive research on the identification of pheromones in Psyllid pests. Pheromones have been reported in four species of psyllids (Hoton et al., 2008; Bazzaz et al., 2019). Male psyllids have been found to be attracted to females, and these pheromones have been identified as sex pheromones. The active components identified include relatively low-volatility long-chain cuticular hydrocarbons (CHCs), such as 13-methylheptacosane, found in the pear psyllid Cacopsylla chinensis (Wan et al., 2013), and highly volatile lauric acid and acetic acid, found in the citrus psyllid Diaphorina citri (Zanardi et al., 2018). However, the presence of pheromones in the wolfberry psyllid and the specific pheromone compounds have not been reported. It is urgent to identify the specific pheromone species and active components of these pheromones in the wolfberry psyllid, thereby developing new monitoring and green control methods based on the pheromones of the wolfberry psyllid to alleviate the problem of high pesticide residues in chemical pest control, which affects the quality of wolfberries. Summary of the Invention

[0004] The purpose of the present invention is to provide a sex pheromone of wolfberry psyllid. The present invention identifies the active component of the sex pheromone of wolfberry psyllid, thereby obtaining a sex attractant of wolfberry psyllid, and develops a monitoring and green control method based on the wolfberry psyllid pheromone, thereby alleviating the problem of high pesticide residues in chemical control that affects the quality of wolfberries.

[0005] The present invention first provides the use of 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, toluene, 2-hexanone and / or 1-methylcyclopentanol as or in the preparation of wolfberry psyllid sex pheromones.

[0006] When used, the 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, toluene, 2-hexanone and / or 1-methylcyclopentanol are preferably prepared into a solution using n-hexane;

[0007] When used, the concentration of the 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, 2-hexanone and / or 1-methylcyclopentanol in the solution is 10 to 10 4 μg / mL, preferably 10 to 5000 μg / mL.

[0008] The present invention further provides a wolfberry psyllid sex attractant, the active ingredients of which are 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, 2-hexanone and / or 1-methylcyclopentanol;

[0009] The active ingredients of the wolfberry psyllid sex attractant are preferably 2,5-dimethyltetrahydrofuran, 2-hexanone and / or 1-methylcyclopentanol.

[0010] Furthermore, the active ingredient of the wolfberry psyllid sex attractant is preferably 2,5-dimethyltetrahydrofuran, 2-hexanone, 1-methylcyclopentanol or a mixture of the three;

[0011] In the mixture, the mass ratio of the 2,5-dimethyltetrahydrofuran, the 2-hexanone and the 1-methylcyclopentanol is 1-2:1-3:1-6.5;

[0012] In the mixture, the mass ratio of the 2,5-dimethyltetrahydrofuran, the 2-hexanone and the 1-methylcyclopentanol is preferably any one of the following:

[0013] 1)1~1.9:1~2.6:1~6.1; 2)1:1:1; 3)1.8:2.6:5.6; 4)1.9:2:6.1.

[0014] Based on the wolfberry psyllid sex attractant, the present invention further provides a wolfberry psyllid sex attractant attractant core, comprising a carrier and the wolfberry psyllid sex attractant loaded on the carrier;

[0015] The carrier is rubber;

[0016] The mass ratio of the wolfberry psyllid sex attractant to the carrier is 1:5000-700000, for example, 1 μg-1000 μg of the wolfberry psyllid sex attractant is loaded on a 0.5-0.7 g rubber attractant carrier.

[0017] The wolfberry psyllid sex attractant lure core can be prepared according to the following steps:

[0018] The wolfberry psyllid sex attractant is prepared into a solution using a solvent (such as n-hexane), and then the carrier is immersed in the solution to obtain the product.

[0019] The use of the wolfberry psyllid sex attractant and the wolfberry psyllid sex attractant core provided by the present invention in monitoring and forecasting, interfering with mating, trapping, killing and / or preventing wolfberry psyllid also falls within the protection scope of the present invention.

[0020] The present invention has the following beneficial effects:

[0021] The present invention extracts, screens, and identifies the active ingredient of a sex pheromone from a lycium arvense extract. Using n-hexane as a solvent, the resulting attractant exhibits a significant indoor attractant effect. When used in conjunction with sticky insect traps in the field, it effectively attracts lycium arvense. The present invention also provides a new method for monitoring and controlling lycium arvense. The discovered active ingredient of the sex pheromone is readily available and low-cost, exhibits a significant attractant effect, and offers advantages such as simplicity, ease of operation, ease of commercialization, and a green, sensitive process. Furthermore, the present method does not cause pesticide residues, can reduce the amount of chemical pesticides required for lycium arvense control, and helps alleviate the problem of high pesticide residues in lycium arvense. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Figure 4 shows the mating and egg-laying rhythms of Lycium barbarum psyllid. Figure A shows the mating rhythm, Figure B shows the egg-laying rhythm, Figure C shows the mating duration, and Figure D shows the egg-laying amount.

[0023] Figure 2 Figure 1 shows the EAG responses of the wolfberry psyllid to body surface volatiles of male and female insects in Example 1 of the present invention. Figure A shows the EAG responses of unmated female insects to body surface volatiles of mated and unmated female and male insects, Figure B shows the EAG responses of unmated male insects to body surface volatiles of mated and unmated female and male insects, Figure C shows the EAG responses of mated female insects to body surface volatiles of mated and unmated female and male insects, and Figure D shows the EAG responses of mated male insects to body surface volatiles of mated and unmated female and male insects.

[0024] Figure 3 Figure 1 shows the behavioral responses of the wolfberry psyllid to live male and female insects in Example 1 of the present invention, wherein Figure A shows the behavioral responses of female insects to live male and female insects, and Figure B shows the behavioral responses of male insects to live male and female insects.

[0025] Figure 4 This is the behavioral response of Lycium barbarum to volatiles from the female insect's body surface in Example 2 of the present invention, wherein white is the control and gray is the female insect's body surface extract.

[0026] Figure 5 Figure 3 shows the GC-EAD response of the wolfberry psyllid to volatiles from the surface of females. Figure A shows the antennal electrophysiological response of male wolfberry psyllids to volatiles from the surface of unmated females. The five locations outlined by dotted lines represent distinct antennal potential responses of the wolfberry psyllids that correspond exactly to the peak times of the substances above. Figure B shows the structures of the five active components identified by GC-MS.

[0027] Figure 6 This is a comparison of volatiles from the body surfaces of male and female Lycium barbarum psyllids in Example 3 of the present invention. Figure A is a GC-MS total ion current chromatogram of volatiles from the body surfaces of female insects, Figure B is a GC-MS total ion current chromatogram of volatiles from the body surfaces of female insects, and Figure C is a comparison of the contents of 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, 2-hexanone, and 1-methylcyclopentanol in male and female insects.

[0028] Figure 7 The field attracting effect of the lure prepared with the active component of the sex pheromone of the wolfberry psyllid in Example 5 of the present invention on the male wolfberry psyllid, wherein CK is the n-hexane control, L1-L18 are lure cores configured in corresponding proportions, and CK+ is the positive control of one female wolfberry psyllid.

[0029] Figure 8 This is the field attracting effect of the lure prepared with the active component of the sex pheromone of the wolfberry psyllid in Example 5 of the present invention on female wolfberry psyllids, wherein CK is the n-hexane control, L1-L18 are lure cores configured in corresponding proportions, and CK+ is the positive control of one female wolfberry psyllid.

[0030] Figure 9 The results show the field attracting effect of the lure prepared with the active component of the sex pheromone of the wolfberry psyllid in Example 5 of the present invention on male and female wolfberry psyllids, wherein CK is a n-hexane control, L1-L18 are lure cores configured in corresponding proportions, and CK+ is a positive control of one female wolfberry psyllid. DETAILED DESCRIPTION

[0031] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0032] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0033] The extraction conditions of the wolfberry psyllid pheromone in the present invention are clarified as follows:

[0034] To observe the mating rhythm of the wolfberry psyllid, the tip of a wolfberry branch was wrapped with wet cotton wool to keep it moist and placed in a petri dish. A newly emerged male and female pair of wolfberry psyllids were collected and placed in the dish. An infrared camera was used to record the psyllids' diurnal mating rhythm and duration for 72 hours. Twelve replicates were used to observe the oviposition rhythm of the wolfberry psyllid. A pot of wolfberry seedlings was placed in a nylon cage (30 × 30 × 40 cm). Three newly emerged male and female pairs of wolfberry psyllids were introduced. The number of eggs on the wolfberry seedlings was checked every hour for 72 hours. Four replicates were used. Daily egg production was observed for a total of 38 replicates. The cage experiment was used to observe the egg production and rhythm of female wolfberry psyllids. A pot of wolfberry seedlings was placed in a nylon cage (30 × 30 × 40 cm). Three pairs of newly emerged, male and female wolfberry psyllids (2, 3, and 4 days old) were placed in the cage and allowed to move freely. Egg production by the wolfberry psyllids on the wolfberry seedlings was checked and recorded every hour after the experiment began. Four or more replicates were performed per day of age.

[0035] Since pheromones are signal substances that regulate the behavior of the wolfberry psyllid, the peak period of pheromone production of the wolfberry psyllid should be related to the peak period of the psyllid's mating behavior. Figure 1 (Figures A and B in the middle) Their mating behavior begins 1 hour after light exposure. In dark conditions, the wolfberry psyllid is inactive, and the psyllid's mating and egg-laying behavior are less frequent. The mating behavior of the wolfberry psyllid mainly occurs during the light period, starting at 8 a.m., with two active peaks at noon and 8 p.m. In addition, the study found that the wolfberry psyllid becomes sexually mature and lays eggs one day after emerging, which clarified the time conditions for the next step of collecting and extracting the wolfberry psyllid's pheromones.

[0036] Example 1: Antennae electrophysiological and behavioral responses of Lycium barbarum to male and female insects

[0037] 1. Experimental Methods

[0038] 1. Test insect source

[0039] The Lycium barbarum psyllids used in this experiment were collected from Yinchuan, Ningxia, and identified morphologically and molecularly. They were reared indoors in the laboratory at a temperature of 26±2°C, a humidity of 60±5%, and a photoperiod of 16 L:8 D. Potted Lycium barbarum seedlings served as food and oviposition substrate. Adults were immediately removed and sexed after eclosion. Males and females were caged separately and used as virgins. Some virgins were placed together in a cage to mate and serve as mateds. All male and female Lycium barbarum psyllids tested were 2-3 days old.

[0040] 2. Collection of volatile substances on the body surface

[0041] Volatiles from the body surface of the wolfberry psyllid were extracted using an extraction method. After being stunned with CO2, the insects were separated into males and females. Three hundred mated and unmated female and male wolfberry psyllids were collected and placed into 2 mL brown sample bottles. The samples were extracted with 1 mL of n-hexane (chromatographic grade, Fisher Scientific) for 10 minutes. The supernatant was collected and stored at -20°C. The sample was concentrated to 500 μL using a nitrogen purge prior to antennal electrophysiological measurements. Four treatments were used: unmated females and males, and mated females and males.

[0042] 3. Antennae electrophysiological responses of Lycium barbarum to volatiles from the body surfaces of male and female insects

[0043] The electrophysiological responses of the wolfberry psyllid to volatiles collected from the bodies of mated and unmated male and female insects were measured using a potentiometer. 10 μL of the collected n-hexane solvent and volatiles from mated and unmated male or female insects were dripped onto a 2×10 cm filter paper strip. After the solution evaporated, the strip was placed in a Pasteur tube and sealed with Parafilm. The Parafilm was removed during the experiment to serve as the test odorant. The head of the wolfberry psyllid was removed, and the distal ends of the antennae were excised with a scalpel. The head was connected to a reference electrode, and the distal ends of the antennae were connected to a recording electrode. A glass electrode was filled with 0.1 mol / L KCl solution as the electrolyte. After the antennae were connected and the baseline stabilized, the test odorant was added for measurement. The pedal was pressed for 0.2 s stimulation, with a minimum of 30 s between stimulations to allow the antennae to stabilize. The response to the n-hexane solvent was measured at the beginning and end of each antennae as a control. Each insect served as a replicate, with 15 biological replicates for each sex. The electrophysiological responses of the antennae were detected by IDAC2 (Syntech, Germany), and the results were recorded and processed by EAGPro 2 software. The differences between the treatment and control groups were tested by Student's t test.

[0044] 4. Behavioral selection of male and female wolfberry psyllids

[0045] A Y-shaped olfactometer was used to observe the behavioral responses of male and female Lycium barbarum psyllids to volatiles produced by both male and female Lycium barbarum psyllids. The Y-shaped glass tube had a diameter of 1 cm, with two 10 cm long arms at a 60° angle. Each arm was connected to a different odor source bottle. The experimental airflow was generated by an atmospheric sampler (300 mL / min, QC-1B, Beijing Labor Protection Institute). The airflow passed sequentially through activated carbon, a distilled water washing bottle, and a gas flowmeter. It was then split through the odor source bottle and finally entered the two arms of the Y-shaped tube. During the experiment, 30 male or female Lycium barbarum psyllids were placed in one arm of the Y-shaped tube as the odor source, while the other arm served as a blank control. After the experiment, all glassware was cleaned with detergent, distilled water, and alcohol, followed by drying at 200°C for 2 hours. The entire apparatus was placed in a behavioral chamber (100 × 90 × 65 cm) illuminated with 5000 lux. The experimental environment was maintained at 26 ± 2°C. During the experiment, the wolfberry psyllids were gently placed one by one at the starting point of the Y-shaped tube, given 1 minute to adapt, and observed for 5 minutes. When the wolfberry psyllid entered 1 / 3 of the length of the arm and stayed there for more than 30 seconds, it was recorded as a choice. Each group of 3 psyllids was divided into 1 group. After each group was tested, the Y-shaped glass tube was reversed 180° to eliminate the influence of position effect on the behavior of the wolfberry psyllids. A new Y-shaped glass tube was replaced after every 2 groups. More than 39 psyllids were tested for each odor, and the behavioral test time was 9:00-22:00. The experimental data results were tested for significance by chi-square test. "*" indicates that the odor source has a significant attraction to the wolfberry psyllids (P < 0.05).

[0046] 2. Experimental Results

[0047] The results of the electrophysiological experiment showed that the mated male insects had a significant electrophysiological response to the extracts from the body surface of the unmated female insects (t 18 =-3.245, p = 0.004), and mated females also showed significant electrophysiological responses to the surface extraction of unmated females (t 28 =-2.119, p = 0.043), unmated male and female insects did not show obvious electrophysiological responses to the odors of mated or unmated male and female insects, such as Figure 2 shown.

[0048] The results of indoor behavioral experiments showed that female Lycium barbarum psyllids were not significantly affected by females (X 2 = 0.471, p = 0.493) or male attraction (X 2 = 0.133, p = 0.715), but males were significantly attracted to females (X 2 = 4.5, p = 0.034), and were not significantly attracted to males (X 2 =0.133, p=0.715), as Figure 3 shown.

[0049] Example 2: Behavioral responses of Lycium barbarum to a mixture of sexual pheromones

[0050] 1. Experimental Methods

[0051] A Y-shaped olfactometer was used to observe the behavioral responses of adult male Lycium barbarum psyllids to volatiles from the body surfaces of female Lycium barbarum psyllids, using the same method as in Example 1. The Y-shaped glass tube had a diameter of 1 cm, with two 10 cm arms at a 60° angle. Each arm was connected to a different odor source bottle. The experimental airflow was generated by an atmospheric sampler (300 mL / min, QC-1B, Beijing Labor Insurance Institute) and passed sequentially through activated carbon, a distilled water wash bottle, and a gas flowmeter. The airflow then passed through the odor source bottle and entered the two arms of the Y-shaped tube. During the experiment, one arm of the Y-shaped tube was filled with 10 μL of a female volatile extract (containing volatiles from approximately three females), while the other arm was filled with a filter paper piece (2 cm × 1 cm) with 10 μL of n-hexane as a blank control. A total of 50 individuals were tested, and behavioral testing was conducted from 9:00 AM to 10:00 PM. The experimental data were tested for significance by chi-square test, and “*” indicates that the odor source has a significant attraction effect on the wolfberry psyllid (P<0.05).

[0052] 2. Experimental Results

[0053] The results of indoor behavioral experiments showed that male insects were significantly attracted to the volatile extracts on the surface of female insects (X 2 =6.737, p=0.009), as Figure 4 shown.

[0054] Example 3: Screening and identification of active substances in the sexual semiochemicals of Lycium barbarum Psyllid

[0055] 1. Experimental Methods

[0056] 1. Collection of volatile substances on the body surface

[0057] Volatiles from the body surface of Lycium barbarum psyllids were extracted using an extraction method. Lycium barbarum psyllids were stunned with CO2 and then sexed. 300 mated and unmated female and male Lycium barbarum psyllids were collected and placed into 2 mL brown sample bottles. The samples were extracted with 1 mL of n-hexane (chromatographic grade, Fisher Scientific) for 10 minutes. The supernatant was collected and stored at -20°C. The sample was concentrated to 100 μL using a nitrogen purifier before injection. Four treatments were set up: unmated females and males, and mated females and males, with six biological replicates per treatment.

[0058] 2. GC-EAD screening of active substances in semiochemicals

[0059] The electrophysiological responses of male antennae of the wolfberry psyllid to volatile samples collected from the female insect's body surface were measured using a GC-EAD coupled device. The GC-FID was an Agilent 7890 gas chromatograph with a flame detector and a DB-5 column model (30 m × 0.25 mm × 0.25 μm). The antennae of male wolfberry psyllids were processed in the same manner as described above (Example 1: Antennae Electrophysiological Response). The column oven temperature program was as follows: hold at 40°C for 2 minutes, then increase to 200°C at a rate of 20°C / min, then increase to 280°C at a rate of 10°C / min and hold for 15 minutes. The carrier gas was nitrogen at a flow rate of 1 mL / min. Data were analyzed using GC / EAD32 software (Syntech), with at least four biological replicates.

[0060] 3. Identification and content comparison of active substances

[0061] Volatiles collected from male and female Lycium barbarum psyllids were separated and identified using an Agilent 7200B GC / Q-TOF (HP-5MS) and a Shimadzu GC-MS QP2020 (RTX-5MS: 30 m × 0.25 mm × 0.25 μm). The inlet temperature was 260°C, the injection volume was 2 μL, the mass spectrometer ionization mode was EI70 eV, and the carrier gas was helium at a flow rate of 1.0 mL / min. The heating program was the same as above. Volatiles were identified by searching the NIST14 database, comparing the differences in volatiles between male and female insects, and identifying active compounds by retention time and mass spectra. These were further confirmed by comparing the retention time and mass spectra of standards with those of the samples. In previous experiments, no trans-2-hexenal was found in the volatile substances on the surface of male and female insects. Therefore, when quantifying the content of active substances, 25 ng of trans-2-hexenal was added to the extracts of volatile substances on the surface of male and female insects (diluted with n-hexane). The active substances in female and male wolfberry psyllids were quantified by the internal standard method (the percentage of target substance to internal standard).

[0062] 2. Experimental Results

[0063] The GC-EAD was used to screen the active components of the pheromone of the wolfberry psyllid in the extract of the wolfberry psyllid. The study found that the male wolfberry psyllid showed obvious electrophysiological responses to various components, such as Figure 5 As shown, the structures of these active compounds were identified by GC-MS and compared with standards. The active ingredients were 2,3-dimethylpentane, 2,5-dimethyltetrahydrofuran, toluene, 2-hexanone, and 1-methylcyclopentanol. Figure 5 shown.

[0064] By comparing the components of the surface extracts of male and female Lycium barbarum psyllids through GC-MS analysis, it was found that there was no significant difference in the types of compound components between the two, such as Figure 6As shown in Figures A and B. By measuring the content of different components in the extracts of male and female body surfaces using the internal standard method, it was found that the content of 2-hexanone in the extracts of female wolfberry psyllids was significantly higher than that in the extracts of male wolfberry psyllids (t 10 =2.85, p=0.017), as Figure 6 As shown in Figure C.

[0065] Example 4: Behavioral responses of Lycium barbarum psyllids to single components of active substances in sexual semiochemicals

[0066] 1. Experimental Methods

[0067] The behavioral responses of male and female Lycium barbarum psyllids to the active ingredient standard (Sigma-Aldrich) were tested using a Y-type olfactometer. The active ingredient standard was diluted to 10 6 ng / mL, 10 5 ng / mL, 10 4 ng / mL, 10 3 ng / mL series gradient, 10 μL of the diluted standard solution was dripped onto a 2 × 2.5 cm filter paper strip as the odor source for behavioral testing. A filter paper strip dripped with 10 μL of n-hexane served as a control. Other procedures were the same as above. The data from this experiment were tested for significance using a chi-square test. At least 39 individuals of each sex and standard concentration were tested. The data were tested for significance using a chi-square test.

[0068] 2. Experimental Results

[0069] Indoor testing of the behavioral responses of male and female adults of Lycium barbarum to the identified active ingredient standards is shown in Table 1. The results show that Lycium barbarum showed obvious behavioral responses to 2,5-dimethyltetrahydrofuran, 2-hexanone, and 1-methylcyclopentanol. 3- 10 5 The attraction rate of 2,5-dimethyltetrahydrofuran at the concentration of ng / mL to both male and female insects exceeded 60%, with female insects showing a strong attraction to 10 5 ng / mL of 2,5-dimethyltetrahydrofuran showed a significant tendency (X 2 =3.903, p=0.048), males vs. 10 5 ng / mL of 2,5-dimethyltetrahydrofuran also showed a significant tendency (X 2 =3.903, p=0.048). For 2-hexanone, 10 3 ng / mL concentration of 2-hexanone and 10 4- 10 5 ng / mL concentration of 2-hexanone attracted both female and male insects by more than 60%, and compared with the control, male wolfberry psyllids attracted 10 4 ng / mL(X 2=4.235, p=0.04) and 10 5 ng / mL(X 2 =3.903, p=0.048) concentration of 2-hexanone has a significant trend. For 1-methylcyclopentanol, 10 4 The attraction rate of 1-methylcyclopentanol to both male and female insects reached more than 60%, but the attraction was not significant, and at 10 6 ng / mL concentration, it was found that female wolfberry insects were repelled by 1-methylcyclopentanol (X 2 =3.903, p=0.048).

[0070] Table 1 Behavioral responses of male and female wolfberry psyllids to single components of active substances in sexual pheromones

[0071]

[0072] Example 5: Field Attraction Effect of Lures Prepared from Active Components of Lycium barbarum Psyllid Sex Pheromone

[0073] 1. Experimental Methods

[0074] Dilute 2,5-dimethyltetrahydrofuran, 2-hexanone, and 1-methylcyclopentanol with n-hexane, and add 20 μL of the mixture according to the ratio shown in the figure (2,5-dimethyltetrahydrofuran single dose, 2-hexanone single dose, 1-methylcyclopentanol single dose, male insect ratio, female insect ratio, 2,5-dimethyltetrahydrofuran: 2-hexanone: 1-methylcyclopentanol = 1:1:1, where the male insect ratio refers to ... A single dose of 2,5-dimethyltetrahydrofuran (DMTF) and 1-methylcyclopentanol (1.9:2.0:6.1, respectively). The female insect ratio (2,5-DMTF, 2-hexanone, 1-methylcyclopentanol) was added to silicone rubber plugs (0.5-0.7 mg) to give a final dose of 1, 10, and 100 μg per lure, respectively. Hexane was used as a control. The prepared lures were stored at 4°C. White plastic sticky traps (20 cm × 25 cm) were used to eliminate the influence of color on the attraction of the wolfberry psyllid. Silicone rubber plugs with different treatments were suspended from the center of the sticky traps using wire and then hung approximately 1.0 m above the wolfberry plants, both upwind and downwind. A row of wolfberry plants formed a block, with only one replicate per block. Treatments within each block were completely randomized, with at least 15 m between treatments and at least 15 m between blocks. A sticky insect plate with a silicone rubber plug containing the solvent n-hexane was used as a negative control, and a sticky insect plate with a female wolfberry psyllid stuck on it was used as a positive control.

[0075] Before the experiment, blank sticky traps without silicone rubber stoppers were hung in the experimental plots. One day later, the number of blank sticky traps in all experimental plots was counted and statistically analyzed to confirm that there were no significant differences in the initial insect populations between plots. Sticky traps with different silicone rubber stoppers were then hung according to the experimental setup to investigate the attraction of different lures prepared with sex pheromone active components to the wolfberry psyllid. Sticky traps and lures were replaced daily, and the position of each treatment was rotated. The number of male and female wolfberry psyllids captured on the traps was recorded and analyzed daily. Each treatment represented a replicate, and the experiment was conducted over four days, for a total of 21 replicates. Because the total number of insects, females, and males attracted did not conform to normal or Poisson distributions, the nonparametric Kruskal-Wallis H test was used to test for significant differences between treatments, and the Mann-Whitney test was used for multiple comparisons between treatments.

[0076] 2. Experimental Results

[0077] like Figure 7 As shown in the results, the number of male psyllids attracted by the 1 μg female insect ratio treatment group (Mann-Whitney test, Z = -3.978, p < 0.001) and the 100 μg 2-hexanone treatment group (Mann-Whitney test, Z = -2.843, p = 0.004) was significantly higher than that of the n-hexane control group. The number of male psyllids attracted by the 1 μg female insect ratio treatment group (Mann-Whitney test, Z = -3.257, p = 0.001) and the 100 μg 2-hexanone treatment group (Mann-Whitney test, Z = -2.035, p = 0.042) was significantly higher than that of the positive control group. In addition, the 1 μg female insect ratio treatment group was significantly higher than that of the 1 μg 2,5-dimethyltetrahydrofuran treatment group (Mann-Whitney test, Z = -2.876, p = 0.004) and the 1 μg male insect ratio treatment group (Mann-Whitney test, Z = -2.8 The results showed that the male-female ratio was 100 μg (Mann-Whitney test, Z=-3.412, p=0.001) and the ratio of males to females was 100 μg (Mann-Whitney test, Z=-2.801, p=0.005). The different baits also showed an attractive effect on female psyllids. Compared with the control, 10 μg of 2,5-dimethyltetrahydrofuran alone, 100 μg of 2-hexanone alone, and the three mixtures attracted more females, with 100 μg of 2-hexanone alone attracting the most females (Mann-Whitney test, Z=-1.992, p=0.046).

[0078] The different treatments of baits significantly affected the total number of Psyllids attracted (Kruskal-Wallis H test, χ 2(19) = 31.875, p = 0.032). Among them, the total number of psyllids attracted by the 1 μg female insect ratio treatment group was the highest (3.76 ± 0.68), which was significantly higher than that of the n-hexane control group (Mann-Whitney test, Z = -4.208, p < 0.001). The total number of psyllids attracted by the 100 μg 2-hexanone treatment group was also significantly higher than that of the n-hexane control group (Mann-Whitney test, Z = -3.743, p < 0.001) ( Figure 9 The total number of psyllids attracted by the 1 μg female insect ratio treatment group (Mann-Whitney test, Z=-3.210, p=0.001) and the 100 μg 2-hexanone treatment group (Mann-Whitney test, Z=-2.614, p=0.009) was also significantly higher than that of the positive control group.

Claims

1. Use of 2,5-dimethyltetrahydrofuran, 2-hexanone, 1-methylcyclopentanol, or a mixture thereof as or in the preparation of a sex pheromone for Lycium barbarum psyllid; In the mixture, the mass ratio of 2,5-dimethyltetrahydrofuran, 2-hexanone and 1-methylcyclopentanol is 1-1.9:1-2.6:1-6.

1.

2. A wolfberry psyllid sex attractant, the active ingredient of which is a mixture of 2,5-dimethyltetrahydrofuran, 2-hexanone, and 1-methylcyclopentanol; In the mixture, the mass ratio of 2,5-dimethyltetrahydrofuran, 2-hexanone and 1-methylcyclopentanol is 1-1.9:1-2.6:1-6.

1.

3. A wolfberry psyllid sex attractant core, comprising a carrier and 2,5-dimethyltetrahydrofuran, 2-hexanone, 1-methylcyclopentanol, or the wolfberry psyllid sex attractant according to claim 2, loaded on the carrier; The carrier is rubber; The mass ratio of the wolfberry psyllid sex attractant to the carrier is 1:5000-700000; The mass ratio of the 2,5-dimethyltetrahydrofuran or the 2-hexanone or the 1-methylcyclopentanol to the carrier is 1:5000-700000.

4. Use of the wolfberry psyllid sex attractant according to claim 2 and the wolfberry psyllid sex attractant core according to claim 3 in monitoring and forecasting, interfering with mating, and trapping wolfberry psyllids.

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