Application of bitter gourd volatile substances in repelling liriomyza trifoliate
By screening the volatile substances of bitter melon trans-2-hexenal and myrtleenol, a repellent was developed for trilephant squid, which solved the problems of chemical control resistance and lack of green prevention and control, and achieved environmentally friendly repellent effects and product development.
Patent Information
- Application Number
- CN202510553101.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-29
- Publication Date
- 2025-08-01
AI Technical Summary
In the prior art, chemical control of trileopard flies has led to an increase in drug resistance and lacks effective green ecological prevention and control methods. The application of volatile substances of bitter melon to avoid trileopard flies has not been reported.
Using volatile substances of bitter melon, including trans-2-hexenal and myrtleenol, through screening of different concentrations and combination ratios, a repellent was developed for green prevention and control of trileaf squids. The dosage forms include powders, sustained release agents, aerosols or sprays.
It provides effective repelling effect on the three-leaf spotted flies, is environmentally friendly, harmless to the human body, has economic and ecological benefits, and supports the development of green ecological prevention and control products and field applications.
Smart Images
Figure CN120391469A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the application of Momordica charantia volatile substances in repelling Liriomyza trifolii, and belongs to the field of biotechnology. Background Art
[0002] Liriomyza trifolii is a world-wide pest that seriously harms horticultural vegetables. In recent years, with the development of protected agriculture, the harm of leafminer pests has become increasingly serious; Liriomyza trifolii has a wide range of hosts, and both its adults and larvae can cause harm to crops. The larvae mine the leaves or petioles of host plants, seriously affecting the photosynthesis of host plants, resulting in defoliation, flower dropping of host plants, delayed development and withering. The adults feed and lay eggs on the leaves, and the damaged wounds of plants also provide a way for pathogenic bacteria to invade.
[0003] At present, chemical control is still the main method for controlling Liriomyza trifolii in production. The frequent and irregular use of a large number of chemical pesticides has led to a gradual increase in its resistance level, greatly reducing the control effect of insecticides. Therefore, in view of the prominent problems of Liriomyza trifolii in agricultural production, it is imperative to explore new prevention and control technologies. At present, the prevention and control of invasive species has shifted from simply eradicating invasive species themselves in the past to ecological pest control based on biodiversity and ecological restoration of invaded and damaged habitats, including the "push-pull" prevention and control strategy using functional plants. Plant volatiles are a type of secondary substances with strong volatility in plants, which are easily detected by insects and have effects such as repelling and attracting insects. Scientific and reasonable utilization of plant volatiles can effectively control pests. In the early stage, the laboratory detected the selective differences of Liriomyza trifolii in kidney beans and different functional plants by using a "Y"-type olfactometer. The results showed that Momordica charantia had a better repellent effect on the adults of Liriomyza trifolii compared with both the air control and the preferred host, but there is no report on the application of Momordica charantia volatiles in Liriomyza trifolii at present. Summary of the Invention
[0004] Object of the Invention: The technical problem to be solved by the present invention is to provide the application of Momordica charantia volatile substances in repelling Liriomyza trifolii, so as to provide a basis for the subsequent development of green ecological control products for Liriomyza trifolii.
[0005] Technical Solution: To solve the above technical problem, the present invention provides the application of Momordica charantia volatile substances in repelling Liriomyza trifolii.
[0006] Among them, the Momordica charantia volatile substances include one or both of trans-2-hexenal and myrtenol.
[0007] Among them, the concentration of trans-2-hexenal is 0.1 - 100 μL / mL.
[0008] Furthermore, the concentration of the trans-2-hexenal is 1 to 100 μL / mL.
[0009] Furthermore, the concentration of the trans-2-hexenal is 1 to 10 μL / mL.
[0010] Among them, the concentration of the myrtenol is 0.1 to 100 μL / mL.
[0011] Furthermore, the concentration of the myrtenol is 1 to 100 μL / mL.
[0012] Furthermore, the concentration of the myrtenol is 1 μL / mL or 100 μL / mL.
[0013] Among them, when the balsam pear volatile substance is a combination of trans-2-hexenal and myrtenol, the volume ratio of trans-2-hexenal to myrtenol is 1 to 9:1 to 9.
[0014] Furthermore, the volume ratio of trans-2-hexenal to myrtenol is 9:1 or 1:9.
[0015] The present invention also provides a Liriomyza trifolii repellent, which contains the balsam pear volatile substance.
[0016] Among them, the balsam pear volatile substance includes one or both of trans-2-hexenal and myrtenol.
[0017] Among them, the dosage form of the Bemisia tabaci repellent includes powder, sustained-release agent, aerosol or spray.
[0018] Among them, the concentration of trans-2-hexenal in the Liriomyza trifolii repellent is 0.1 to 100 μL / mL.
[0019] Furthermore, the concentration of trans-2-hexenal in the Liriomyza trifolii repellent is 1 to 100 μL / mL.
[0020] Furthermore, the concentration of trans-2-hexenal in the Liriomyza trifolii repellent is 1 to 10 μL / mL.
[0021] Among them, the concentration of myrtenol in the Liriomyza trifolii repellent is 0.1 to 100 μL / mL.
[0022] Furthermore, the concentration of myrtenol in the Liriomyza trifolii repellent is 1 to 100 μL / mL.
[0023] Furthermore, the concentration of myrtenol in the Liriomyza trifolii repellent is 1 μL / mL or 100 μL / mL.
[0024] Among them, when the balsam pear volatile substances are a combination of trans-2-hexenal and myrtenol, the volume ratio of trans-2-hexenal to myrtenol is 1-9:1-9.
[0025] Through the identification and screening of the volatile substances of the balsam pear, a plant that repels Liriomyza trifolii, and the verification of different combinations and pairings of standard volatile substance products, the volatile substance standard products, concentrations, and combination ratios with the best repellent effects were obtained. In this experiment, different concentrations of volatile substances were set at 0.1 μL / mL, 1 μL / mL, 10 μL / mL, and 100 μL / mL. Finally, it was determined that the repellent effect of trans-2-hexenal and myrtenol substances at a concentration of 1 μL / mL with a ratio of 9:1 was the best.
[0026] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages: The method for identifying, screening, and combined application of the Liriomyza trifolii repellent involved in the present invention can provide a basis for the development and field application of green ecological control products for Liriomyza trifolii. It is environmentally friendly, harmless to humans, and has good economic and ecological benefits. Description of the Drawings
[0027] Figure 1 It is the behavioral response of Liriomyza trifolii of the present invention to different repellent plants;
[0028] Figure 2 It is the GC-MS result peak diagram of the balsam pear volatile substances of the present invention;
[0029] Figure 3 It is the behavioral response of Liriomyza trifolii of the present invention to different concentrations of trans-2-hexenal, trans-3-hexen-1-ol, and myrtenol;
[0030] Figure 4 It is the directional behavioral response of Liriomyza trifolii of the present invention to different ratios of a combination of 2 volatile substances. Detailed Embodiments
[0031] The technical solution of the present invention will be further described below in conjunction with the drawings.
[0032] Example 1 Identification and Screening of Balsam Pear Volatile Substances
[0033] (1) Test the behavioral response of Liriomyza trifolii to repellent plants (garlic leaves, scallion leaves, coriander, Chinese chives, balsam pears), and measure it using a "Y"-type olfactometer. Before the test, ventilate the olfactometer for 15 minutes to eliminate the deviation of other odors from the test, and the air flow rate is 200 mL / min.
[0034] A healthy host plant (Phaseolus vulgaris) and different repellent plants were placed in two odor source bottles respectively. Subsequently, Liriomyza trifolii was introduced into the straight arm of the "Y"-tube, and observations were made 10 minutes later. Liriomyza trifolii that moved more than 1 / 3 of the length in the two branch arms of the "Y"-tube was recorded as a choice. After each test, the "Y"-tube olfactometer was cleaned with absolute ethanol and then heated to 100 °C, with ventilation maintained for 5 min. The positions of the odor source bottles to the arms were periodically reversed to eliminate directional bias. A pure air control group was also established in the experiment. In each group of tests, 10 newly emerged adults were placed in one tube, and each tube of insects only participated in one test. Every 3 tests were used as one technical replicate, and there were 3 technical replicates in total. All determinations were carried out at a temperature of 25 ± 0.5 °C and a relative humidity of 70 ± 5%. One-way analysis of variance and Tukey's method were used for multiple comparisons. Among them, *** indicates a significant difference between the treatment and the control at P < 0.001, ** indicates a significant difference at P < 0.01, and * indicates a significant difference at P < 0.05 level.
[0035] As Figure 1 shown, the behavioral responses of Liriomyza trifolii to repellent plants were tested using a "Y"-tube olfactometer. Momordica charantia had the best repellent effect on Liriomyza trifolii (the selection rates of repellent plants compared with the host plant were: garlic leaves 28.89%, scallion leaves 36.67%, coriander 33.33%, Chinese chives 46.67%, and Momordica charantia 23.33%; the selection rates of repellent plants compared with air were: garlic leaves 26.67%, scallion leaves 27.78%, coriander 31.11%, Chinese chives 45.56%, and Momordica charantia 21.11%).
[0036] (2) Identification of Momordica charantia volatiles: Momordica charantia leaves were collected, and the volatile components were identified using SPME-GC-MS technology to determine its main components. According to the identification results, the components with significant repellent effects on Liriomyza trifolii were screened out.
[0037] Among them, the GC conditions were: DB-WAX fused silica capillary column (30 m × 0.25 mm, 0.25 μm); the carrier gas was helium with a flow rate of 1.0 mL / min; the septum purge flow rate was 3.0 mL / min; the injection port temperature was 230 °C; splitless injection; the temperature program was an initial temperature of 40 °C held for 3 min, then increased to 150 °C at a rate of 6 °C / min and held for 1 min, and finally increased to 220 °C at a rate of 10 °C / min and held for 3 min. The MS conditions were: the interface temperature was 230 °C; the ion source temperature was 230 °C; the quadrupole temperature was 150 °C; the ionization mode was electron impact (EI) with an electron energy of 70 eV; the mass scan range (m / z) was 35 - 500.
[0038] Determination process: Collect bitter gourd leaves, cut the bitter gourd leaves from the stem with scissors, put the leaves into a test bottle, seal the bottle mouth with tin foil, and preheat the test bottle in a water bath at 60 °C. Before extraction, age the adsorption fiber extraction head of the instrument device 50 / 30um DVB / CAR / PDMS at 250 °C for 30 min. After aging, carefully penetrate the SPME device syringe needle through the tin foil, while avoiding the fiber head contacting the bottle wall and the test material. Then carefully and slowly push the handle to gradually extend the fiber head in the syringe needle for adsorption, and the adsorption time is 40 min. After extraction is completed, perform thermal desorption on the machine for 7 min. At this time, let the GC-MS enter the state to be measured. After adsorption is completed, push the handle to retract the extraction head into the handle, carefully withdraw the SPME device from the bottle, immediately insert it into the GC injection port, and desorb for 7 min. The GC-MS is processed by TurboMass Ver 6.1.2, and the separated peak patterns are retrieved through the NIST 17.0 spectral library.
[0039] The GC-MS result peak pattern is as Figure 2 shown. Further analyze the volatile components of bitter gourd leaves.
[0040] Table 1 Volatile components of bitter gourd leaves
[0041]
[0042] As shown in Table 1, the relative contents of trans-2-hexenal (34.015%), trans-3-hexen-1-ol (21.396%), and myrtenol (7.906%) are higher than those of other substances.
[0043] Investigation on the repellent effect of volatile standards in Example 2
[0044] Select 3 kinds of volatile standards with relatively high contents (trans-2-hexenal, trans-3-hexen-1-ol, myrtenol), use liquid paraffin as the solvent, and dilute the standards into 4 concentration gradients of 0.1 μL / mL, 1 μL / mL, 10 μL / mL, and 100 μL / mL respectively. Take liquid paraffin as the control, and use a "Y"-type olfactometer to observe the behavioral responses of Liriomyza trifolii to different standards and different concentrations.
[0045] Before the experiment, the olfactometer was ventilated for 15 min to eliminate the deviation of other odors from the test, and the air flow rate was 200 mL / min. The required standard concentration was prepared with liquid paraffin. 20 μL of the sample to be tested and 20 μL of liquid paraffin were respectively pipetted and dropped onto a qualitative filter paper of 2 cm × 2 cm and placed in the odor source bottle of the "Y"-type olfactometer. Subsequently, Liriomyza trifolii was placed in the straight arm of the "Y"-type tube and observed after 10 min. The "Y"-type olfactometer was placed in the dark to reduce visual interference, and an LED light source was used to provide stable illumination; Liriomyza trifolii that moved more than 1 / 3 of the length in the two branched arms of the "Y"-type tube was recorded as a choice. After each test, the "Y"-type olfactometer was cleaned with absolute ethanol and then heated to 100 °C and ventilated for 5 min. The position from the odor source bottle to the arm was periodically reversed to eliminate the direction deviation. In each group of tests, 10 newly emerged adults were placed in one tube, and each tube of insects only participated in one test. Every 3 tests were used as a technical replicate, and there were 3 technical replicates in total. All determinations were carried out at a temperature of 25 ± 0.5 °C and a relative humidity of 70 ± 5%. Then, the data was analyzed according to the method of Example 1.
[0046] As Figure 3 shown, (E)-2-hexenal at concentrations of 1 μL / mL (selection rate: 11.11%), 10 μL / mL (selection rate: 14.44%), and 100 μL / mL (selection rate: 27.78%) and myrtenol at concentrations of 1 μL / mL (selection rate: 12.22%) and 100 μL / mL (selection rate: 23.33%) had significant repellent effects on Liriomyza trifolii, while (E)-3-hexen-1-ol had no significant repellent effect on Liriomyza trifolii at 4 different concentrations.
[0047] Example 3 Combined Application of Volatile Standards
[0048] The optimal repellent concentrations of 1 μL / mL (E)-2-hexenal and 1 μL / mL myrtenol were set at volume ratios of 1:1, 9:1, and 1:9 respectively, and the "Y"-type olfactometer was used to observe the selection behavior responses of Liriomyza trifolii under different combinations of volatile standards.
[0049] Before the experiment, the olfactometer was aerated for 15 min to eliminate the deviation of other odors from the test, and the air flow rate was 200 mL / min. The stock solutions of trans-2-hexenal at 1 μL / mL and myrtenol at 1 μL / mL were prepared with liquid paraffin. The ratios of 1:1, 9:1, and 1:9 required for the test were prepared with the stock solutions. 20 μL of the sample to be tested and 20 μL of liquid paraffin (control group) were pipetted and dropped onto qualitative filter papers of 2 cm × 2 cm respectively, and placed in the odor source bottles of the "Y"-type olfactometer. Subsequently, Liriomyza trifolii were introduced into the straight arm of the "Y"-type tube, and observations were made 10 min later. Liriomyza trifolii that moved more than 1 / 3 of the length in the two branched arms of the "Y"-type tube were recorded as selections. After each test, the "Y"-type olfactometer was cleaned with absolute ethanol and then heated to 100 °C and ventilated for 5 min. The positions from the odor source bottles to the arms were periodically reversed to eliminate directional deviation. In each group of tests, 10 newly emerged adults were placed in one tube, and each tube of insects only participated in one test. Every 3 tests were used as a technical replicate, and there were 3 replicates in total. All determinations were carried out at a temperature of 25 ± 0.5 °C and a relative humidity of 70 ± 5%. Data analysis was performed according to the method of Example 1.
[0050] As Figure 4 shown, trans-2-hexenal at 1 μL / mL and myrtenol at 1 μL / mL had significant repellent effects on Liriomyza trifolii at the ratios of 9:1 (selection rate: 10.00%) and 1:9 (selection rate: 18.89%). Among them, the selection rate was the lowest at 9:1, and it had the strongest repellent effect on Liriomyza trifolii.
Claims
1. Application of bitter gourd volatile substances in repelling Liriomyza trifolii.
2. The application according to claim 1, characterized in that, The bitter gourd volatile substances include one or both of trans-2-hexenal and myrtenol.
3. The application according to claim 2, characterized in that The concentration of trans-2-hexenal is 0.1 - 100 μL / mL.
4. The application according to claim 2, wherein The concentration of myrtenol is 0.1 - 100 μL / mL.
5. The application according to claim 2, wherein When the bitter gourd volatile substances are a combination of trans-2-hexenal and myrtenol, the volume ratio of trans-2-hexenal to myrtenol is 1 - 9:1 - 9.
6. A Liriomyza trifolii repellent, characterized in that, It contains bitter gourd volatile substances.
7. The Liriomyza trifolii repellent according to claim 6, characterized in that, The bitter gourd volatile substances include one or both of trans-2-hexenal and myrtenol.
8. The Liriomyza trifolii repellent according to claim 7, characterized in that, The concentration of trans-2-hexenal is 0.1 - 100 μL / mL.
9. The Liriomyza trifolii repellent according to claim 7, wherein The concentration of myrtenol is 0.1 - 100 μL / mL.
10. The Liriomyza trifolii repellent according to claim 7, characterized in that, When the bitter gourd volatile substances are a combination of trans-2-hexenal and myrtenol, the volume ratio of trans-2-hexenal to myrtenol is 1 - 9:1 - 9.