Application of citrus volatile matter in repelling pardosa pseudoannulata

Abstract

The invention discloses application of citrus volatile matter to repelling pardosa pseudoannulata, and belongs to the technical field of spider repelling. The invention provides an application of a citrus volatile matter in repelling pardosa pseudoannulata, and provides an application of the citrus volatile matter in preparing a pardosa pseudoannulata repellent, an application of the citrus volatile matter alpha-terpilenol in repelling pardosa pseudoannulata and an application of the citrus volatile matter alpha-terpilenol in preparing the pardosa pseudoannulata repellent based on the application. According to the invention, alpha-terpilenol with a repelling effect on pardosa pseudoannulata is used for developing the spider repellent, so that the spider repellent has great development potential and important significance.

Description

Technical Field

[0001] The invention belongs to the technical field of spider repelling, and particularly relates to use of citrus volatiles in repelling Pardosa pseudoannulata. Background Art

[0002] Pardosa pseudoannulata ( & Strand, 1906), belongs to the order Araneae, family Lycosa. It is distributed in Guizhou, Henan, Shaanxi, Hebei, Beijing, Shanghai, Gansu, Jilin, Liaoning, Jiangxi, Yunnan, etc.

[0003] The Pardosa pseudoannulata has a ferocious habit and a terrifying appearance. Although it is a predatory natural enemy, people still hope to stay away from it in daily life. Currently, the common methods of repelling spiders are physical repelling or spraying chemical agents. The former is ineffective and passive, and the latter is prone to pollution and toxicity.

[0004] At present, the development of a repellent for Pardosa pseudoannulata has not been reported, and there is no report in the art on the use of citrus volatiles to repel Pardosa pseudoannulata. Summary of the invention

[0005] In order to solve the technical problem of filling the above-mentioned gap in the prior art in the field, the present invention provides the use of citrus volatiles to repel Pardosa pseudoannulata.

[0006] The technical solution of the present invention is as follows:

[0007] Use of citrus volatiles to repel Pardosa pseudoannulata.

[0008] The citrus volatiles are: α-terpineol.

[0009] Use of citrus volatiles in preparing a repellent for Pardosa pseudoannulata.

[0010] The Pardosa pseudoannulata repellent contains citrus volatiles as active ingredients;

[0011] Preferably, the citrus volatiles are: α-terpineol.

[0012] Use of citrus volatile α-terpineol to repel Pardosa pseudoannulata.

[0013] The effective dosage of citrus volatile α-terpineol to repel Pardosa pseudoannulata is 10-100 μL.

[0014] Application of citrus volatile alpha-pineneol in preparing Pardosa pseudoannulata repellent.

[0015] The annular Pardosa spider repellent contains citrus volatile α-terpineol as an active ingredient;

[0016] Preferably, the Pardosa annularis repellent further comprises auxiliary materials.

[0017] The beneficial effects of the present invention are as follows:

[0018] The present invention unexpectedly discovered that citrus volatiles: α-terpineol has a repellent effect on the Pseudoannularis pardosa spider. The Pseudoannularis pardosa spider's preference response to α-terpineol was tested through a choice preference behavior experiment, and the electrophysiological response of α-terpineol to the Pseudoannularis pardosa spider's sensor was tested through EAG (insect antennal potential), verifying the repellent effect of α-terpineol. The behavioral test results showed that α-terpineol had a very significant (P < 0.001) repellency to the Pseudoannularis pardosa spider in the behavioral test.

[0019] The present invention also further tests and obtains the concentration gradient of citrus extract α-pinene alcohol and the repellent effect of the volatile mixture, screens out the most suitable repellent concentration, and is mainly used for indoor repellent of Pardosa pseudoannulata. Its significance lies in: there are widespread spider phobia patients in the world, and they feel a sense of fear when they see spiders or come into contact with spiders at ordinary times, and even fear being in a closed environment where they think there may be spiders (such as the appearance of spider webs, etc., which remind people of spiders). After the fear of spiders is stimulated, some people's symptoms will last for several days, during which they will constantly suspect that there are spiders in the environment around them, causing trouble to normal life. Using the repellent effect of α-pinene alcohol on spiders, spider repellents can be made, which can be used to drive spiders away from specific environments on the one hand, and can relieve the anxiety of patients with spider phobia on the other hand, so spider repellents have greater development potential and are of great significance. The present invention reflects the behavioral activity of Pardosa pseudoannulata to citrus volatiles: α-pinene alcohol through indoor behavioral selection experiments and antenna potential experiments, and proves that α-pinene alcohol has a repellent effect on Pardosa pseudoannulata. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a physical device diagram of the two-way selection box system used in Section 1.2.1 of Experimental Example 1 of the present invention.

[0021] Figure 2 This is a physical device diagram of the glass electrode used in Section 1.2.2 of Experimental Example 1 of the present invention.

[0022] Figure 3 This is a bar graph showing the selectivity of Pardosa pseudoannulata to citrus volatiles in Section 2.1 of Experimental Example 1 of the present invention.

[0023] Figure 4 This is a bar graph of the potential values ​​of the antennal potential response of Pardosa pseudoannulata to citrus volatiles in Section 2.2 of Experimental Example 1 of the present invention.

[0024] Figure 5This is a bar chart of the number of webs built by the Pardosa pseudoannulata in the field in the treatment group and the control group in Section 2.3 of Experimental Example 1 of the present invention. DETAILED DESCRIPTION

[0025] The contents of the present invention are further described in detail below in conjunction with specific embodiments and experimental examples, but the protection scope of the present invention is not limited thereto.

[0026] Group 1 Example: Use of citrus volatiles to repel Pardosa pseudoannulata

[0027] This group of embodiments provides the use of citrus volatiles to repel Pardosa pseudoannulata.

[0028] In a specific embodiment, the citrus volatiles are: α-terpineol.

[0029] The second group of examples: Use of citrus volatiles in preparing a repellent for Pardosa pseudoannulata

[0030] This group of embodiments provides the use of citrus volatiles in preparing a repellent for Pardosa pseudoannulata.

[0031] In a specific embodiment, the Pardosa pseudoannulata repellent has citrus volatiles as active ingredients;

[0032] Preferably, the citrus volatiles are: α-terpineol.

[0033] The third group of examples: Use of citrus volatile α-terpineol to repel Pardosa pseudoannulata

[0034] This group of embodiments provides the use of citrus volatile α-pineneol to repel Pardosa pseudoannulata.

[0035] In a specific embodiment, the effective amount of citrus volatile α-terpineol for repelling Pardosa pseudoannulata is 10-100 μL.

[0036] Example 4: Use of citrus volatile α-terpineol in preparing a repellent for Pardosa pseudoannulata

[0037] This group of embodiments provides the use of citrus volatile α-pineneol in preparing a repellent for Pardosa pseudoannulata.

[0038] In a specific embodiment, the Pardosa annularis repellent has citrus volatile α-terpineol as an active ingredient;

[0039] Preferably, the Pardosa annularis repellent further comprises auxiliary materials.

[0040] Experimental Example 1

[0041] 1 Materials and methods

[0042] 1.1 Experimental materials and treatments

[0043] Collection and feeding of experimental spiders: Pardosa pseudoannulata of the genus Pardosa of the family Lycosa was collected from lawns all over the campus of Jinggangshan University and raised individually in a transparent plastic box (length × width × height = 16.5 cm × 11.5 cm × 5.8 cm). Sterile cotton balls soaked in water were placed in the box. The temperature in the laboratory was set at 25 ± 2 ° C, and the photoperiod was 14L: 10D (i.e., 14 hours of daylight and 10 hours of darkness). Young spiders or adult spiders of advanced age were selected for the experiment. They were not fed for two days before the experiment to keep them hungry.

[0044] Extraction of citrus volatiles: Citrus volatiles were extracted by organic solvent extraction for the test. Five newly grown citrus leaves were picked with alcohol-wiped tweezers and placed in a 10 mL clean glass bottle. The organic solvent dichloromethane was poured in. After 5 minutes of extraction, the leaves were taken out and the solution in the bottle was dried and concentrated with a nitrogen blower until about 1 mL of concentrated solution remained for EAG and behavioral tests.

[0045] Reagents and instruments: α-Terpineol (purity ≥ 95%), TCI (Shanghai) Chemical Industry Development Co., Ltd., Japan.

[0046] The electroantennography (EAG) instrument consisted of a stimulus airflow controller (CS-55, Syntech, the Netherlands) and a signal recording controller (Keysight 34461A, Tangshan Dinggan Technology Co., Ltd.).

[0047] 1.2 Research Methods

[0048] 1.2.1 Potential antennal effect of Pardosa pseudoannulata on citrus volatile α-terpineol

[0049] Studies have confirmed that spiders have taste hairs on their bodies that can sense chemicals in the environment, usually on the tarsus and metatarsus of their legs. Xiao Yonghong and others used antennal electroencephalography (EAG) (Syntech, the Netherlands) to successfully record the potential response of the tarsus and metatarsus of the female Beijing ghost spider to the sex pheromones of male spiders.

[0050] During the experiment, the tarsal segment of the first step of the Pardosa pseudoannulata was selected, and the claw at the end was cut off. The two ends of the walking leg were quickly connected to two homemade electrodes (a glass capillary was made into a pointed shape with a wire drawer, and a conductive silver wire and physiological saline were placed inside it, see Figure 2, so that both ends of the walking leg are fully in contact with the physiological saline. The other ends of the two glass electrodes are connected to the antenna potential electrode. The potential response of the receptors on the feet is amplified by the DG3FA-1 high-impedance microelectrode amplifier (Tangshan Dinggan Technology Co., Ltd.) and displayed on the computer. Drop the test volatiles on a clean filter paper (length × width = 2CM × 1CM, Taizhou Jinao Paper Co., Ltd.) and place it at the air flow outlet. During the experiment, the stimulation was performed every 3 minutes. After each substance was stimulated, a new filter paper was replaced to stimulate the next substance. The air test was performed 6 minutes before the start, after each group of tests, and 6 minutes before the end of the test of the walking leg. Each experiment lasted 33 minutes, with a total of 10 repetitions. In the experiment, the continuous humidification gas flow was set to 250mL / min, the stimulation gas flow was 200mL / min, and the stimulation method was foot-operated pulse delivery.

[0051] 1.2.2 Experiment on the selective behavior of Pardosa pseudoannulata to the citrus volatile α-terpineol

[0052] The spider behavior choice test device uses a two-way choice box system ( Figure 1 ), which was used to test the choice preference behavior of Pireneitega luctuosa and Hololena curta

[19] The two-way choice box system is composed of three transparent polypropylene plastic boxes side by side (top 16.5cm×11.5cm, bottom 14cm×8.5cm, box height 5.8cm), with two choice boxes on the left and right, and a release box in the middle. The release box is connected to the left and right choice boxes on both sides with a circular hole of about 4cm in diameter. The test spider can freely choose to enter and exit the left and right choice boxes in the release box. During the experiment, the left and right choice boxes were randomly set as treatment boxes (with the test agent) and control boxes (without the test agent). In order to facilitate the spider to climb in the box, scratches were made on the inner walls of the three boxes by hand to make them rough. The two-way choice box system was used to test the preference response of Pardosa pseudoannulata to α-terpineol. The test environment was a closed behavior room with low airflow, an ambient temperature of 24-27℃, a relative humidity of 50-60%, and α-terpineol was diluted 10 times with dichloromethane for use.

[0053] Before the experiment, the spiders to be tested were introduced into the release box, and the connection ports of the selection boxes on both sides were closed to allow them to adapt for 20 minutes. During the experiment, the reagent (treatment group: α-pineneol, control group: dichloromethane) was dropped on the qualitative filter paper (length × width = 3CM × 3CM, Taizhou Jinao Paper Co., Ltd.) at a dose of 10ul, and after evaporation for 2 minutes, it was placed in the selection box, and the connection ports on both sides of the selection box were opened, and the experiment began. During the experiment, records were recorded every 20 minutes, and the experiment lasted for 2 hours. After each test of a volatile substance, the filter paper was replaced, and the two-way selection box was wiped with 95% anhydrous ethanol, and the next volatile substance test was carried out after air drying. The sample size of each experimental group was 40 spiders.

[0054] 1.2.3 Effects of citrus volatile α-terpineol on outdoor web site selection by Pardosa pseudoannulata

[0055] The large lawn in the north of Jinggangshan University campus was selected as the test site, where a large number of P. pseudoannularis spiders live all year round. During the experiment, the reagent (α-terpineol for the treatment group and dichloromethane for the control group) was dropped into the rubber lure at a dose of 50ul and placed at the roots of the lawn. Each group was repeated 5 times. The area within a radius of 3 meters with the lure as the dot was used as the test area. The original P. pseudoannularis spider webs in this area were destroyed, and the number of spider webs re-woven in the area was counted three days later.

[0056] 1.3 Data Analysis

[0057] Chi-square test was used to analyze the differences between the treatment group and the control group in the behavior selection experiment, and one-way analysis of variance (ANOVA) was used to analyze the differences between the peak values ​​of EAG responses of each reagent. The significant value P<0.05 was significant, and P<0.001 was extremely significant. All data statistical analyses were completed by Excel and SPSS19.0 (IBM), and the drawings were completed by Origin2018 (OringinLab) drawing software.

[0058] 2 Results and analysis

[0059] 2.1 Electroantennographic responses of Pardosa pseudoannulata to citrus volatiles and α-terpineol

[0060] The peak values ​​of the EAG potential responses of P. pseudoannulata to citrus volatiles and α-terpineol are shown in Table 1. The data were compared between groups using one-way analysis of variance (ANOVA) and the least significant difference (LSD) method. Figure 4 ) showed that the potential response values ​​of the receptors on the legs of Pardosa pseudoannulata to citrus volatiles and α-terpineol were higher than those of the air and dichloromethane controls (P < 0.05).

[0061] Table 1. Peak antennal potentials of Pardosa pseudoannulata in response to citrus volatiles

[0062]

[0063] 2.2 Indoor choice behavior of Pardosa pseudoannulata to the citrus volatile α-terpineol

[0064] In this study, a two-way choice box system was used to study the preference of Pardosa pseudoannulata for α-terpineol, an extract from citrus fruit. The statistical analysis results showed that the number of times Pardosa pseudoannulata entered the treatment box containing α-terpineol was significantly less than the number of times it entered the control box (χ 2 =17.065, df=1, P<0.001, Figure 3 ). This indicates that α-terpineol has a very significant repellent effect on Pardosa pseudoannulata.

[0065] 2.3 Effects of citrus volatile α-terpineol on site selection and web-building behavior of P. pseudoannulata in the wild

[0066] After the web of the Pardosa pseudoannulata is destroyed, it will quickly build a new web as its main habitat. When it senses that there are factors in the environment that are not conducive to its habitat, it may abandon the original location where it wove its web and choose to weave a web in another location. In the experiment, in an area with a radius of 3 meters and α-terpineol as the center, the experimental results are as follows: Figure 5 As shown in the figure, the number of webs of the control group was 18.40±4.56, and the number of webs of the treatment group was 3.80±1.64, with a significant difference between the groups (t=6.734, df=8, P<0.001). It can be seen that the Pseudoannularis spider tends not to build webs and live in areas with citrus volatile α-terpineol, indicating that α-terpineol has a repellent effect on the Pseudoannularis spider within a range of 3 meters.

Claims

1. Use of citrus volatiles to repel Pardosa pseudoannulata.

2. The use of citrus volatiles to repel Pardosa pseudoannulata according to claim 1, characterized in that: The citrus volatiles are: α-terpineol.

3. Use of citrus volatiles in preparing a repellent for Pardosa pseudoannulata.

4. The use of citrus volatiles for preparing a Pardosa pseudoannulata repellent according to claim 3, characterized in that: The Pardosa pseudoannulata repellent contains citrus volatiles as active ingredients; Preferably, the citrus volatiles are: α-terpineol.

5. Use of citrus volatile α-pineneol to repel Pardosa pseudoannulata.

6. The use of citrus volatile α-terpineol for repelling Pardosa pseudoannulata according to claim 5, characterized in that: The effective concentration of citrus volatile α-terpineol to repel Pardosa pseudoannulata is 10-100 μL.

7. Use of citrus volatile α-pineneol in preparing a repellent for Pardosa pseudoannulata.

8. Use of the citrus volatile α-terpineol in preparing a repellent for Pardosa pseudoannulata according to claim 7, characterized in that: The annular Pardosa spider repellent contains citrus volatile α-terpineol as an active ingredient; Preferably, the Pardosa annularis repellent further comprises auxiliary materials.

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