Application of 4-fluorophenylacetaldehyde and compositions containing same in attracting cotton bollworm

Abstract

The present invention relates to the use of 4-fluorophenylacetaldehyde and a composition containing the same in attracting cotton bollworms. The present invention finds that 4-fluorophenylacetaldehyde has a significant attracting effect on cotton bollworms, so it can be developed into an attractant for attracting cotton bollworms to achieve the purpose of controlling cotton bollworms.

Description

Technical Field

[0001] The invention relates to the field of attractants, in particular to application of 4-fluorophenylacetaldehyde and a composition containing the same in attracting cotton bollworms. Background Art

[0002] cotton bollworm Helicoverpa armigera The cotton bollworm is a major global agricultural pest, hosting over 200 plant species. Currently, control of the cotton bollworm relies primarily on chemical pesticides, but their widespread use has led to environmental pollution, ecological damage, increased pest resistance, and potential harm to non-target organisms. Furthermore, pests like the cotton bollworm are prone to developing resistance to chemical pesticides, leading to a gradual decline in control effectiveness and increasing both the cost and difficulty of control.

[0003] Insect behavior modulators are widely used in the environmentally friendly control of agricultural pests. Developed based on insect olfaction, these agents precisely control pest behavior by mimicking or interfering with chemical communication through specialized odor compounds. Due to their high sensitivity and specificity, odor compounds effectively attract or repel pests, are less likely to induce pest resistance, and do not cause long-term ecosystem contamination. They also have minimal impact on non-target organisms during use, helping to maintain ecological balance.

[0004] Therefore, screening highly effective compounds with attractant effects is of great significance for the green prevention and control of pests such as cotton bollworm. Summary of the Invention

[0005] One aspect of the present invention provides the use of p-fluorophenylacetaldehyde in attracting cotton bollworms.

[0006] The second aspect of the present invention provides an attractant composition, which includes p-fluorophenylacetaldehyde and a pesticide-acceptable carrier.

[0007] In one embodiment, the amount of p-fluorophenylacetaldehyde is 0.1 to 50 μg.

[0008] In one embodiment, the amount of p-fluorophenylacetaldehyde is 1 to 50 micrograms.

[0009] In one embodiment, the amount of p-fluorophenylacetaldehyde is 20 to 50 micrograms.

[0010] In one embodiment, the amount of p-fluorophenylacetaldehyde is 21 to 46 micrograms.

[0011] In a specific embodiment, the attractant composition further comprises butyl salicylate, eucalyptol, ALPHA-pinene, 4-methoxybenzyl alcohol and limonene.

[0012] In a specific embodiment, based on the total mass of p-fluorophenylacetaldehyde, butyl salicylate, eucalyptol, ALPHA-pinene, 4-methoxybenzyl alcohol and limonene as 100%, the amount of p-fluorophenylacetaldehyde is 18% to 24%, the amount of butyl salicylate is 22% to 26%, the amount of eucalyptol is 10% to 20%, the amount of ALPHA-pinene is 18% to 24%, the amount of 4-methoxybenzyl alcohol is 8% to 15%, and the amount of limonene is 5% to 10%.

[0013] In a specific embodiment, based on the total mass of p-fluorophenylacetaldehyde, butyl salicylate, eucalyptol, ALPHA-pinene, 4-methoxybenzyl alcohol and limonene as 100%, the amount of p-fluorophenylacetaldehyde is 21%, the amount of butyl salicylate is 24%, the amount of eucalyptol is 16%, the amount of ALPHA-pinene is 21%, the amount of 4-methoxybenzyl alcohol is 11%, and the amount of limonene is 7%.

[0014] In a specific embodiment, the attractant composition further comprises methyl 2-methoxybenzoate and isopentanol.

[0015] In a specific embodiment, based on the total mass of p-fluorophenylacetaldehyde, methyl 2-methoxybenzoate and isoamyl alcohol as 100%, the amount of p-fluorophenylacetaldehyde is 40% to 50%, the amount of methyl 2-methoxybenzoate is 20% to 25%, and the amount of isoamyl alcohol is 30% to 35%.

[0016] In a specific embodiment, based on the total mass of p-fluorophenylacetaldehyde, methyl 2-methoxybenzoate and isoamyl alcohol as 100%, the amount of p-fluorophenylacetaldehyde is 46%, the amount of methyl 2-methoxybenzoate is 22%, and the amount of isoamyl alcohol is 32%.

[0017] The third aspect of the present invention provides the use of the attractant composition described in the second aspect of the present invention in attracting cotton bollworms.

[0018] Beneficial effects of the present invention: The present invention discovered that 4-fluorophenylacetaldehyde has a significant attractant effect on cotton bollworms, so it can be developed into an attractant for attracting cotton bollworms to achieve the purpose of preventing and controlling cotton bollworms. Furthermore, 4-fluorophenylacetaldehyde has the following significant advantages: the attractant effect is significant, and it can still show significant attractant activity to cotton bollworms at an extremely low dose of 0.1 micrograms. The development of such attractant compounds can not only reduce the use of chemical pesticides, but also effectively protect natural enemy insects and promote agricultural ecological safety. Furthermore, the composition containing fluorophenylacetaldehyde is also more attractive to cotton bollworms. In general, 4-fluorophenylacetaldehyde and its composition have the advantages of significant cotton bollworm attractant effect, environmental friendliness, not easy to produce resistance and broad application prospects, and are of great significance for the green prevention and control of pests such as cotton bollworms. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Shown are the electrophysiological responses of cells heterologously expressing the cotton bollworm odorant receptor protein to the tested odor compounds.

[0020] Figure 2 The selection behavior of cotton bollworm on phenylacetaldehyde is shown.

[0021] Figure 3 The selection behavior of cotton bollworm on p-fluorophenylacetaldehyde is shown.

[0022] Figure 4 The selective behavior of cotton bollworm on 2,3-difluorophenylacetaldehyde is shown.

[0023] Figure 5 The selective behavior of cotton bollworm on 3,4-difluorophenylacetaldehyde is shown.

[0024] Figure 6 The selective behavior of cotton bollworm on 3,4,5-trifluorophenylacetaldehyde is shown.

[0025] Figure 7 The selective behavior of cotton bollworm on 2-(2,3,4-trifluorophenyl)acetaldehyde is shown.

[0026] Figure 8 Shown is the selection behavior of cotton bollworm on 2-(2,4,5-trifluorophenyl)acetaldehyde.

[0027] Figure 9 A comparison of the choice behavior of cotton bollworm on phenylacetaldehyde and p-fluorophenylacetaldehyde is shown.

[0028] Figure 10 The field selection of the p-phenylacetaldehyde attractant composition and the p-fluorophenylacetaldehyde attractant composition of cotton bollworm in Example 4 is shown.

[0029] Figure 11 The field selection of the p-phenylacetaldehyde attractant composition and the p-fluorophenylacetaldehyde attractant composition of cotton bollworm in Example 5 is shown. DETAILED DESCRIPTION

[0030] The above contents of the present invention are further described in detail below in the form of preferred implementation cases, but they do not constitute a limitation of the present invention.

[0031] Unless otherwise specified, the materials and reagents in the examples of the present invention can be purchased through commercial channels.

[0032] The compound used in the present invention is p-fluorophenylacetaldehyde, whose English name is (4-Fluoro-phenyl)-acetaldehyde and whose structural formula is as follows: .

[0033] The compound 2,3-difluorophenylacetaldehyde used in the present invention has the following structural formula: .

[0034] The compound 3,4-difluorophenylacetaldehyde used in the present invention has the English name (3,4-Difluorophenyl)acetaldehyde and has the following structural formula: .

[0035] The compound 3,4,5-trifluorophenylacetaldehyde used in the present invention has the English name (3,4,5-Trifluorophenyl)acetaldehyde and has the following structural formula: .

[0036] The compound 2,3,4-trifluorophenylacetaldehyde used in the present invention has the following structural formula: .

[0037] The compound 2,4,5-trifluorophenylacetaldehyde used in the present invention has the following structural formula: .

[0038] Phenylacetaldehyde was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0039] 4-Fluorophenylacetaldehyde was purchased from J&K Technology Co., Ltd.

[0040] 2,3-Difluorophenylacetaldehyde, 3,4-difluorophenylacetaldehyde, 3,4,5-trifluorophenylacetaldehyde, 2,3,4-trifluorophenylacetaldehyde and 2,4,5-trifluorophenylacetaldehyde were purchased from Zhengzhou Alpha Chemical Co., Ltd.

[0041] Example 1: Electrophysiological Response Activity of Cells Heterologously Expressing an Odor Receptor Protein of Helicoverpa armigera to Detected Odor Compounds

[0042] Phenylacetaldehyde, p-fluorophenylacetaldehyde, 2,3-difluorophenylacetaldehyde, 3,4-difluorophenylacetaldehyde, 3,4,5-trifluorophenylacetaldehyde, 2-(2,3,4-trifluorophenyl)acetaldehyde and 2-(2,4,5-trifluorophenyl)acetaldehyde were prepared into stock solutions with a concentration of 1 mol / L using dimethyl sulfoxide (DMSO).

[0043] 10× Ringer: NaCl 56.1 g, KCl 1.5 g, MgCl2·6H2O 10.2 g, HEPES 11.9 g, adjust the pH to 7.6 with NaOH, dilute to 1 L with sterile ultrapure water, and filter.

[0044] Tetracycline, streptomycin, gentamicin, and sodium pyruvate were prepared into stock solutions of 50 mg / mL, 100 mg / mL, 10 mg / mL, and 275 mg / mL, respectively, using sterile ultrapure water.

[0045] 100× CaCl2 stock solution: Weigh 4.41 g CaCl2·2H2O and dilute to 500 mL with distilled water to make a 60 mM CaCl2 solution. Filter and set aside.

[0046] Xenopus oocyte washing solution: 100 mL of 10× Ringer, 1 mL of gentamicin stock solution, dilute to 1 L with sterile ultrapure water, filter and set aside.

[0047] Xenopus oocyte culture medium: 100 mL of 10× Ringer medium, 10 mL of 100× CaCl2, 50 mL of horse serum, 1 mL of tetracycline stock solution, 1 mL of streptomycin stock solution, and 2 mL of sodium pyruvate stock solution. Add sterile ultrapure water to 1 L and filter.

[0048] Perfusion buffer (1×Ringer): 100 mL of 10×Ringer, 1 mL of gentamicin stock solution, and 10 mL of 100×CaCl2 stock solution. Add sterile ultrapure water to 1 L and filter.

[0049] Before testing, the stock solution of the odor compound to be tested was diluted to 1.0 × 10 -4 mol / L to prepare the solution of the odor compound to be tested.

[0050] African Xenopus oocytes were used to heterologously express the odorant receptors HarmOR42 and HarmOrco of cotton bollworm (Guo M, Du L, Chen Q et al. Odorant Receptors for Detecting Flowering Plant Cues Are Functionally Conserved across Moths and Butterflies. Mol Biol Evol. 2021 Apr13;38(4):1413-1427). Then, a two-electrode voltage clamp system was used to measure the current value of the African Xenopus oocytes in response to the test solution of the odorant compound to be screened. An equal volume of RNase-free ultrapure water was injected into the African Xenopus oocytes as a negative control. The operation was as follows: mature female African Xenopus was selected, the Xenopus oocytes were dissected and isolated, and the isolated Xenopus oocytes were washed with Xenopus oocyte wash solution to obtain clean Xenopus oocytes (stage V–VII). Among them, the Xenopus oocytes isolated from the same Xenopus were from the same batch. Equal volumes of a 2 μg / μl solution of HarmOR42 cRNA and a 2 μg / μl solution of HarmOrco cRNA were mixed and injected into Xenopus oocytes. A negative control was injected with 27.6 nL of RNase-free ultrapure water. The injected oocytes were placed in Xenopus oocyte culture medium and incubated at 18°C ​​for 3 days. The oocyte responses to the test odorant compounds were then measured using a two-electrode voltage clamp (OC-725 Coocyteclamp, Warner Instruments, Hamden, CT) as follows: The oocytes were first flushed with perfusion buffer. During the measurement, the perfusion buffer was turned off and the test odorant solution was turned on. The Digidata 1440A recorded the response for 15 s. After 15 s of recording, the test compound solution was turned off and the perfusion buffer was turned on. Each biological replicate consisted of five or six Xenopus oocytes from the same batch, and three biological replicates were performed. The recorded data were read by Digidata 1440A software and statistically analyzed using pCLAMP 10.2 software to determine the starting current value (i.e., the data measured when flushing with perfusion buffer) and the maximum current value (i.e., the data measured when flushing with the test compound solution). The difference between the two was the electrophysiological response value of the test odor compound, i.e., the current value. Among them, the current value generated by the negative control group was 0. The current values ​​(i.e., the standardized response values) of the oocytes in response to each test odor compound were statistically analyzed using the one-way ANOVA statistical method and plotted using GraphPad Prism 9.5.1. The results are shown in Figure 2. Figure 1 shown.

[0051] Figure 1 The results showed that HarmOR42 protein could be activated by phenylacetaldehyde, p-fluorophenylacetaldehyde, 2,3-difluorophenylacetaldehyde, 3,4-difluorophenylacetaldehyde, 3,4,5-trifluorophenylacetaldehyde, 2-(2,3,4-trifluorophenyl)acetaldehyde and 2-(2,4,5-trifluorophenyl)acetaldehyde. Among them, there was no significant difference between the fluorine-containing test compounds and phenylacetaldehyde.

[0052] Example 2: Selection behavior of cotton bollworm towards test odor compounds

[0053] 4-Fluorophenylacetaldehyde, 2,3-difluorophenylacetaldehyde, 3,4-difluorophenylacetaldehyde, 3,4,5-trifluorophenylacetaldehyde, 2-(2,3,4-trifluorophenyl)acetaldehyde and 2-(2,4,5-trifluorophenyl)acetaldehyde were prepared with n-hexane to prepare test odor compound solutions with concentrations of 0.01, 0.10 and 1.00 μg / μL, respectively.

[0054] The behavioral selection responses of male and female cotton bollworm adults to the tested odor compounds were determined using a Y-shaped tube.

[0055] The Y-tube consisted of a colorless, transparent glass tube with a diameter of approximately 2.5 cm. The main arm was 14 cm long, and the two arms were 27 cm long, with a 75-degree angle between the arms. The Y-tube was placed flat on a tabletop, with the main arm proximal to the operator and the two side arms distal to the operator. A 5W red light was placed midway between the two side arm outlets as a light source. A QC-1B air sampler (Beijing Institute of Labor Protection) served as the airflow dynamics system. The air flowing from the air sampler was first filtered through activated carbon, then humidified by a humidifier bottle, and finally passed through a flow meter and an odor source to the two arms of the Y-tube. The air flow rate in both arms of the Y-tube was 0.4 L / min. All devices were connected by silicone tubing. The room temperature for the behavioral experiments was 25 ± 1°C, and the humidity was 55% ± 5%.

[0056] Before the experiment begins, adjust the airflow speeds on both arms to ensure consistency. For each compound test sample, test at least 50 three-day-old male and female bollworms, four hours into the dark phase. Record at least 25 selected insects, one at a time. After testing five insects, swap the positions of the experimental group (test compound) and the blank control (n-hexane) to eliminate position effects. Replace the Y-shaped tube with a clean one after every ten insects tested. After the experiment, rinse the Y-shaped tube with water and dry it in a 180°C oven for 2 hours.

[0057] 10 μl of the odor compound solution to be tested was dropped onto a 1×1 cm filter paper. The filter paper was placed in one arm of the Y-shaped tube, and 10 μl of n-hexane was placed on the other arm as a blank control. An adult cotton bollworm was placed headfirst into the main arm of the Y-shaped tube, and the insect's choice was recorded within 5 minutes. If the cotton bollworm entered a side arm and exceeded 1 / 3 of the side wall length and remained there for 1 minute or more, it was considered that the insect had made a clear choice of the odor source connected to that side arm. Otherwise, it was recorded as no choice. The selection rate was calculated after statistics. The results are shown in the table. Figures 2 to 8 .

[0058] Figures 2 to 8 The results showed that phenylacetaldehyde could significantly attract male and female adults of cotton bollworm at 1 microgram, and female cotton bollworm at 10 micrograms; p-fluorophenylacetaldehyde could significantly attract male and female adults of cotton bollworm at 0.1 to 10 micrograms; while 2,3-difluorophenylacetaldehyde, 3,4-difluorophenylacetaldehyde, 3,4,5-trifluorophenylacetaldehyde, 2-(2,3,4-trifluorophenyl)acetaldehyde and 2-(2,4,5-trifluorophenyl)acetaldehyde did not show the effect of attracting male and female adults of cotton bollworm.

[0059] Example 3: Comparison of the selection behavior of cotton bollworms on phenylacetaldehyde and p-fluorophenylacetaldehyde

[0060] Phenylacetaldehyde and p-fluorophenylacetaldehyde were respectively prepared with n-hexane to prepare a solution of the odor compound to be tested with a concentration of 0.10 μg / μl.

[0061] Phenylacetaldehyde was placed in one arm of the Y-shaped tube; p-fluorophenylacetaldehyde was placed in the other arm of the Y-shaped tube. Other operations were the same as in Example 2 to compare the selection behavior of cotton bollworms for phenylacetaldehyde and p-fluorophenylacetaldehyde. Figure 9 .

[0062] Figure 9 The results showed that the attraction effect of 4-fluorophenylacetaldehyde on male and female adults of cotton bollworm was significantly higher than that of phenylacetaldehyde.

[0063] Example 4: Comparison of the selection behavior of cotton bollworms on compositions containing p-fluorophenylacetaldehyde or phenylacetaldehyde

[0064] 21 μg of p-fluorophenylacetaldehyde, 24 μg of butyl salicylate, 16 μg of eucalyptol, 21 μg of ALPHA-pinene, 11 μg of 4-methoxybenzyl alcohol, 7 μg of limonene and 100 μl of solvent paraffin oil were mixed evenly to prepare a p-fluorophenylacetaldehyde attractant composition.

[0065] 21 micrograms of phenylacetaldehyde, 24 micrograms of butyl salicylate, 16 micrograms of eucalyptol, 21 micrograms of ALPHA-pinene, 11 micrograms of 4-methoxybenzyl alcohol, 7 micrograms of limonene and 100 microliters of solvent paraffin oil were mixed evenly to prepare a phenylacetaldehyde attractant composition.

[0066] The attractant experiment was carried out in the soybean fields of Baoding, Hebei from September to October 2024: 20 microliters of 4-fluorophenylacetaldehyde attractant composition and 20 microliters of phenylacetaldehyde attractant composition were respectively added to the cotton roll attractant core, and then the cotton roll attractant cores with each attractant composition were placed in breathable mesh bags and numbered and recorded. The cotton roll attractant core in the mesh bag was placed in the center of the barrel trap, and 5 soybean plots sown at the same time, with the same growth and non-adjacent positions were selected. Each plot was 2 mu, and 3 4-fluorophenylacetaldehyde attractant composition traps and 3 phenylacetaldehyde attractant composition traps were evenly placed in each plot. When placed, they were scattered and numbered randomly. That is, 5 repetitions were set, and 1 plot was 1 repetition. From the 4th day after placement, the number of trapped cotton bollworm adults was investigated and recorded every 3 days, and the investigation was continued until the 11th day. The effect of attracting cotton bollworms in the field was statistically analyzed, and the results are shown in the table. Figure 10 .

[0067] from Figure 10 It can be seen that the number of cotton bollworm adults attracted by the 4-fluorophenylacetaldehyde attractant composition is significantly higher than that by the phenylacetaldehyde attractant composition.

[0068] Example 5: Comparison of the selection behavior of cotton bollworms on compositions containing p-fluorophenylacetaldehyde or phenylacetaldehyde

[0069] 46 μg of p-fluorophenylacetaldehyde, 22 μg of methyl 2-methoxybenzoate, 32 μg of isopentanol and 100 μl of solvent paraffin oil were mixed evenly to prepare a p-fluorophenylacetaldehyde attractant composition.

[0070] 46 μg of phenylacetaldehyde, 22 μg of methyl 2-methoxybenzoate, 32 μg of isopentanol and 100 μl of solvent paraffin oil were mixed evenly to prepare a phenylacetaldehyde attractant composition.

[0071] The attractant experiment was carried out in the soybean fields of Baoding, Hebei from September to October 2024: 20 microliters of 4-fluorophenylacetaldehyde attractant composition and 20 microliters of phenylacetaldehyde attractant composition were respectively added to the cotton roll attractant core, and then the cotton roll attractant cores with each attractant composition were placed in breathable mesh bags and numbered and recorded. The cotton roll attractant core in the mesh bag was placed in the center of the barrel trap, and 5 soybean fields sown at the same time, with the same growth and non-adjacent positions were selected. Each field was 2 mu, and 3 4-fluorophenylacetaldehyde attractant composition traps and 3 phenylacetaldehyde attractant composition traps were evenly placed in each field. When placed, they were scattered and numbered randomly. That is, 5 repetitions were set, and 1 field was 1 repetition. Starting from the 4th day after placement, the number of trapped cotton bollworm adults was investigated and recorded every 3 days, and the investigation was continued until the 11th day. The effect of attracting cotton bollworms in the field was statistically analyzed, and the results are shown in the table. Figure 11 .

[0072] from Figure 11 It can be seen that the number of cotton bollworm adults attracted by the 4-fluorophenylacetaldehyde attractant composition is significantly higher than that by the phenylacetaldehyde attractant composition.

Claims

1. Use of p-fluorophenylacetaldehyde in attracting cotton bollworm, wherein the amount of p-fluorophenylacetaldehyde used is 0.1 to 50 micrograms.

2. An attractant composition comprising p-fluorophenylacetaldehyde and a pesticidal acceptable carrier, wherein the amount of p-fluorophenylacetaldehyde is 0.1 to 50 micrograms.

3. The attractant composition according to claim 2, characterized in that The attractant composition also includes butyl salicylate, eucalyptol, alpha-pinene, 4-methoxybenzyl alcohol and limonene.

4. The attractant composition according to claim 3, characterized in that Based on the total mass of p-fluorophenylacetaldehyde, butyl salicylate, eucalyptol, ALPHA-pinene, 4-methoxybenzyl alcohol and limonene as 100%, the amount of p-fluorophenylacetaldehyde is 18% to 24%, the amount of butyl salicylate is 22% to 26%, the amount of eucalyptol is 10% to 20%, the amount of ALPHA-pinene is 18% to 24%, the amount of 4-methoxybenzyl alcohol is 8% to 15%, and the amount of limonene is 5% to 10%.

5. The attractant composition according to claim 3, characterized in that Based on the total mass of p-fluorophenylacetaldehyde, butyl salicylate, eucalyptol, ALPHA-pinene, 4-methoxybenzyl alcohol and limonene as 100%, the amount of p-fluorophenylacetaldehyde is 21%, the amount of butyl salicylate is 24%, the amount of eucalyptol is 16%, the amount of ALPHA-pinene is 21%, the amount of 4-methoxybenzyl alcohol is 11%, and the amount of limonene is 7%.

6. The attractant composition according to claim 2, characterized in that The attractant composition also includes methyl 2-methoxybenzoate and isoamyl alcohol.

7. The attractant composition according to claim 6, characterized in that Based on the total mass of p-fluorophenylacetaldehyde, methyl 2-methoxybenzoate and isoamyl alcohol as 100%, the amount of p-fluorophenylacetaldehyde is 40% to 50%, the amount of methyl 2-methoxybenzoate is 20% to 25%, and the amount of isoamyl alcohol is 30% to 35%.

8. The attractant composition according to claim 6, characterized in that Based on the total mass of p-fluorophenylacetaldehyde, methyl 2-methoxybenzoate and isoamyl alcohol as 100%, the amount of p-fluorophenylacetaldehyde is 46%, the amount of methyl 2-methoxybenzoate is 22%, and the amount of isoamyl alcohol is 32%.

9. Use of the attractant composition according to any one of claims 2 to 8 in attracting cotton bollworm.

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