Peach tree borer adult attractant and use thereof

Abstract

The application discloses a peach red-necked longicorn adult attractant and application thereof. In particular, the application discloses the use of hexacosane in the preparation of the peach red-necked longicorn attractant. The peach red-necked longicorn adult attractant is hexacosane n-hexane solution with a concentration of 2.5-5 micrograms / microliter. The application first discovers that hexacosane has an attracting effect on female and male adult peach red-necked longicorn, and the preparation of the attractant effectively solves the problems of high labor cost, influence on fruit quality and environmental pollution caused by artificial killing of adult insects, punching injection or drug mud smearing for the prevention and treatment of larvae, thereby providing a practical pollution-free monitoring and prevention and treatment technical means for the monitoring of the occurrence period and amount of adult insects in the forest or large-scale trapping and killing.

Description

Technical Field

[0001] This invention relates to the field of longhorn beetle pest control technology, specifically to an attractant for adult peach red-necked longhorn beetle and its application. Background Technology

[0002] The peach-necked longhorn beetle (Aromia bungii) belongs to the order Coleoptera and the family Cerambycidae. It is a significant trunk-boring pest that damages fruit trees such as peach, apricot, plum, and apricot, as well as ornamental trees such as poplar and willow, throughout China. Due to the concealed nature of the Cerambycidae trunk-boring beetle larvae, it has attracted widespread attention worldwide.

[0003] Currently, the control of the peach red-necked longhorn beetle is mainly carried out through physical and chemical methods. Physical methods such as attracting insects, manual removal, and manual capture have some control effects but are labor-intensive and inefficient. Chemical methods, such as injecting pesticides through holes or applying pesticide mud to control larvae, have potential impacts on fruit quality and are highly toxic, posing health risks. With increasing awareness of agricultural product quality and safety, biological control is a safer, more environmentally friendly, and effective method for pest control. Finding active substances from plant or insect metabolites that regulate pest behavior has gradually become a research hotspot. Using plant-derived attractants for pest control and monitoring has advantages such as high specificity, high efficiency, low resistance development, and no pollution. Therefore, suitable substances with effective attractant activity against the peach red-necked longhorn beetle will greatly contribute to its green control. Summary of the Invention

[0004] The purpose of this invention is to provide an attractant for adult peach-necked longhorn beetles and its application.

[0005] To achieve the objective of this invention, in a first aspect, this invention provides an attractant for adult peach-necked longhorn beetles, wherein the attractant contains hexacosane.

[0006] Furthermore, the attractant is a hexadecane solution prepared with n-hexane as the solvent.

[0007] Preferably, the concentration of the hexadecane solution in the attractant is 2.5–5 μg / μL.

[0008] Secondly, the present invention provides the application of the attractant in attracting and controlling adult peach red-necked longhorn beetles.

[0009] Thirdly, the present invention provides the use of hexadecane in the preparation of attractants for the peach-necked longhorn beetle.

[0010] By employing the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0011] Using plant-derived attractants for pest control and monitoring has advantages such as high specificity, high efficiency, low resistance, and no pollution. This invention is the first to discover that hexadecane has an attractant effect on both male and female adults of the peach red-necked longhorn beetle. Making it into an attractant effectively solves the problems of high labor costs for manual killing and the potential impact on fruit quality caused by drilling holes for injection or applying medicated mud to control larvae. Thus, it provides a practical and pollution-free monitoring and control technology for monitoring the occurrence period and quantity of adult beetles in the forest or for large-scale trapping. Attached Figure Description

[0012] Figure 1 The behavioral response of male and female adult peach-necked longhorn beetles to larval excrement is shown in a preferred embodiment of the present invention.

[0013] Figure 2 The behavioral response of the female peach-necked longhorn beetle to the larval excrement extract is shown in a preferred embodiment of the present invention.

[0014] Figure 3 The behavioral response of male peach-necked longhorn beetle to larval excrement extract is shown in a preferred embodiment of the present invention.

[0015] Figure 4 This is a total ion chromatogram of the extract of the excrement of the peach-necked longhorn beetle larvae in a preferred embodiment of the present invention. ①, ②, and ③ represent hexacosane, 1-heptacosanol, and octadecanaldehyde, respectively.

[0016] Figure 5 The behavioral response of a female peach-necked longhorn beetle to different concentrations of hexacosane is shown in a preferred embodiment of the present invention.

[0017] Figure 6 The behavioral response of male longhorn beetles to different concentrations of hexacosane is shown in a preferred embodiment of the present invention.

[0018] Figure 7 The behavioral response of a female peach-necked longhorn beetle to different concentrations of hexadecaneol is shown in a preferred embodiment of the present invention.

[0019] Figure 8 The behavioral response of male longhorn beetles to different concentrations of hexadecaneol is shown in a preferred embodiment of the present invention.

[0020] Figure 9 The behavioral response of a female peach-necked longhorn beetle to different concentrations of octadecaldehyde is shown in a preferred embodiment of the present invention.

[0021] Figure 10 The behavioral response of male longhorn beetles to different concentrations of octadecaldehyde is shown in a preferred embodiment of the present invention. Detailed Implementation

[0022] The present invention aims to provide a substance, hexacosane, that has an attraction effect on the peach red-necked longhorn beetle, and to provide an attractant containing hexacosane, in order to solve the problems of high cost, time and labor for manual killing of the peach red-necked longhorn beetle due to the lack of effective attraction methods in the current control process, as well as the problems of adverse effects of chemical pesticide control on fruit quality and the ecological environment.

[0023] The present invention adopts the following technical solution:

[0024] This invention provides an attractant for adult peach-necked longhorn beetles, wherein the attractant contains hexacosane.

[0025] Furthermore, the attractant for adult peach-necked longhorn beetles described in this invention is a hexane solution of hexadecane with a concentration of 2.5–5 μg / μL.

[0026] The present invention also provides the use of hexadecane in the preparation of attractants for the peach-necked longhorn beetle.

[0027] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0028] Examples 1-14: Study on attractants for adult peach-necked longhorn beetles

[0029] The excrement of *Rhizoctonia solani* larvae was soaked in hexane (chromatographic grade) at five concentration gradients: 5g, 10g, 20g, 30g, and 50g of excrement were placed in 150mL of hexane (Examples 1-5, respectively), and soaked at 4°C for 24 hours. The supernatant was aspirated with a dropper, and the excrement residue was filtered through glass wool. The extract was stored at 4°C for later use.

[0030] The total ion composition of volatile components in the branches and leaves of the host plant, peach, was determined by GC-MS. Figure 4 The main chemical components (relatively high content) and their relative contents are shown in Table 1: hexacosane; 1-heptacosanol; octadecaldehyde (Examples 6-8, respectively). Hexane solutions with concentrations of 1 μg / μL, 2.5 μg / μL, and 5 μg / μL were prepared. Experiments were conducted (Examples 6-14, respectively) to screen the key substances in the extract of the excrement of the peach-necked longhorn beetle larvae and the optimal concentration of these substances.

[0031] Test insect source:

[0032] The adult beetles used in the Y-type olfactory instrument experiment were collected directly from wild peach orchards during the peak emergence period of the peach red-necked longhorn beetle. All adult beetles used on the day of the experiment were captured the previous day, and the beetles were active and of consistent vitality. They were raised indoors at room temperature (28±3℃).

[0033] I. Behavioral Response Test

[0034] Experiment 1: The behavioral responses of male and female adult peach-necked longhorn beetles to larval feces were tested using a Y-type olfactory instrument. The results are as follows: Figure 1 As shown.

[0035] Experiment 2: The behavioral responses of male and female adult peach-necked longhorn beetles to different concentrations of larval excrement extract were tested using a Y-type olfactory instrument. The results are as follows: Figure 2 , Figure 3 As shown.

[0036] Experiment 3: Gas chromatography-mass spectrometry (GC-MS) was used to identify substances in the excrement extract of the peach-necked longhorn beetle larvae. The results are as follows: Figure 4 The total ion chromatogram and Table 1 are shown.

[0037] Experiment 4: A Y-type olfactory instrument was used to test the behavioral responses of male and female adult peach-necked longhorn beetles to chemical components in larval excrement extracts of different concentrations. The results are as follows: Figures 5-10 As shown.

[0038] Depend on Figure 1 , Figure 2 and combined Figure 3 It can be seen that the extract of the larvae of the peach-necked longhorn beetle at a concentration of 20g / 150mL has a good attraction effect on the peach-necked longhorn beetle.

[0039] Depend on Figure 4 The identification results in Table 1, combined with... Figure 5 , Figure 6 The test results show that hexadecane has the effect of attracting the red-necked longhorn beetle. The hexane solution of hexadecane at a concentration of 2.5-5 μg / μL has a good attraction effect on the red-necked longhorn beetle and can be used to prepare the red-necked longhorn beetle attractant.

[0040] Table 1. Main chemical composition of the excrement extract of the larvae of the peach-necked longhorn beetle.

[0041]

[0042] Example Data Analysis:

[0043] II. Behavioral Response Test

[0044] The response of adult peach-necked longhorn beetles to different concentrations of insect excrement extracts and their main components was tested using a Y-type olfactometer.

[0045] Test Environment: The Y-shaped tube was made of glass, with a main arm length of 40cm and two side arms lengths of 25cm each, forming a 90° angle between the arms. The inner diameter of the tube was 9cm. A spherical glass trap (approximately 2000ml in volume, with a 10cm connection point on each side) was placed at the tip of each side arm of the Y-shaped tube. This trap allowed adult beetles to fall into it when they crawled to it, preventing them from climbing back and thus capturing the pink-necked longhorn beetle moving through the Y-shaped olfactory testing instrument. Air from an atmospheric sampler (1.5L / min) was filtered through activated carbon, passed through the odor source, and finally reached the side tube walls. All devices were connected using Teflon tubing. The Y-shaped olfactory testing instrument was placed indoors, with the two arms facing a natural light source at an intensity of approximately 300–500 lux. Filter paper (cut into 2cm × 2cm squares, manufactured by Sinopharm Chemical Reagent Beijing Co., Ltd.) was used as the odor carrier. Ten μl of insect excrement extract solutions of different concentrations were added to a 2 cm × 2 cm filter paper as the experimental group, and ten μl of hexane solvent was added to another filter paper as the control. The two odor carriers were placed in a spherical glass trap.

[0046] Test conditions: This invention tested the responses of male or female individuals to different concentrations of insect excrement extract and its main components. Clean air flowed through each arm at a rate of 750 mL / min. During the bioassay, a single female or male adult *Rhizophora stylosa* was introduced into the main arm of the Y-type olfactory testing instrument. Each *Rhizophora stylosa* was observed for a maximum of 5 minutes. A selection was considered made when the insect passed halfway through the arm or entered the trap. If no selection was made within the specified time, the insect was excluded from the statistical analysis. Each insect was tested only once, and each group of experiments required at least 30 responding adult *Rhizophora stylosa*. After every 5 insects tested, the experimental and control arms were exchanged to eliminate the position effect on the behavioral test, and new chemical filter paper was used. Before each experiment, all components of the Y-type olfactory instrument were cleaned with anhydrous ethanol, rinsed with distilled water, and dried with hot air. The behavioral bioassay was conducted at 20–25°C and a relative humidity of 45%–50%. Before testing, each adult insect should be placed in a PE box measuring 18×10.5×18cm and starved for 24 hours (26±0.5℃, relative humidity 55%±5%).

[0047] Behavioral tropism analysis was performed using IBM SPSS Statistics (25) (P<0.05), and graphs were generated using Origin 2018. First, the raw data underwent methodological analysis and normality testing. Statistical analysis used a bivariate nonparametric Wilcoxon test to determine whether adult males and females of the peach-necked longhorn beetle selected larval excrement, excrement extracts of different concentrations, and the compounds contained in the extracts.

[0048] Test 1: The odor source for the experimental arm was 10 μL of the attractant prepared in Exercise 1 (5 g / 150 mL of larval excrement extract) used in the Y-type olfactometer experiment, and the control arm was 10 μL of n-hexane.

[0049] The results showed that 36 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 23 chose the experimental arm and 13 chose the control arm (Z = -1.667, P = 0.096, P > 0.05). 41 male adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 26 males chose the experimental arm and 15 chose the control arm (Z = -1.718, P = 0.087, P > 0.05).

[0050] Test 2: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 2 (10 g / 150 mL of larval excrement extract) for a Y-type olfactometer experiment, and the control arm was 10 μL of n-hexane.

[0051] The results showed that 32 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 23 chose the experimental arm and 9 chose the control arm (Z = -2.475, 0.01 < P = 0.013 < 0.05). 33 male adult *Rhododendron simsii* were also used in the Y-type olfactometer experiment, of which 23 males chose the experimental arm and 10 chose the control arm (Z = -2.475, 0.01 < P = 0.013 < 0.05).

[0052] Test 3: The odor source for the experimental arm was 10 μL of the attractant (20 g / 150 mL larval excrement extract) prepared in Example 3, tested using a Y-type olfactometer. The control arm was 10 μL of n-hexane.

[0053] The results showed that 35 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 25 chose the experimental arm and 10 chose the control arm (Z = -2.535, 0.01 < P = 0.011 < 0.05). 34 male adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 25 males chose the experimental arm and 9 chose the control arm (Z = -2.744, 0.001 < P = 0.006 < 0.01).

[0054] Test 4: The odor source for the experimental arm was 10 μL of the attractant (30 g / 150 mL larval excrement extract) prepared in Example 4, tested using a Y-type olfactometer. The control arm was 10 μL of n-hexane.

[0055] The results showed that 33 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 22 chose the experimental arm and 11 chose the control arm (Z = -1.915, P = 0.056, P > 0.05). 36 male adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 25 males chose the experimental arm and 11 chose the control arm (Z = -2.333, 0.01 < P = 0.02 < 0.05).

[0056] Test 5: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 5 (50 g / 150 mL of larval excrement extract) used in the Y-type olfactometer experiment, and the control arm was 10 μL of n-hexane.

[0057] The results showed that 35 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 20 chose the experimental arm and 15 chose the control arm (Z = -0.845, P = 0.398, P > 0.05). 37 male adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 24 males chose the experimental arm and 13 chose the control arm (Z = -1.808, P = 0.071, P > 0.05).

[0058] Based on the above test results and Figure 2 / Figure 3 It was found that the concentration of the extract of the larvae of the peach-necked longhorn beetle (20 g / 150 ml) was most effective in attracting both females and males. Therefore, when identifying the chemical components in the larval excrement by GC-MS, an extract with a concentration of 20 g / 150 mL was selected. High concentrations of excrement were less effective. Based on our long-term experience with Y-type olfactory bioassay experiments, this is because high concentrations easily fill the experimental equipment with the attractant odor, causing adult longhorn beetles to lose their sense of direction in the space where the odor is diffused.

[0059] Test 6: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 6 (a hexane solution of 1 μg / μL hexacosane) for the Y-type olfactometer experiment, and the control arm was 10 μL of hexane.

[0060] The results showed that 32 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 17 chose the experimental arm and 15 chose the control arm (Z = -0.354, P = 0.724, P > 0.05). 31 male adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 17 males chose the experimental arm and 14 chose the control arm (Z = -0.539, P = 0.590, P > 0.05).

[0061] Test 7: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 7 (a hexane solution of 2.5 μg / μL hexacosane) for the Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0062] The results showed that 31 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 19 chose the experimental arm and 12 chose the control arm (Z = -1.257, P = 0.209, P > 0.05). 36 male adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 24 males chose the experimental arm and 12 chose the control arm (Z = -2.000, 0.01 < P = 0.046 < 0.05).

[0063] Test 8: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 8 (a hexane solution of 5 μg / μL hexacosane) for the Y-type olfactory instrument experiment. The control arm was 10 μL of hexane.

[0064] The results showed that 36 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 24 chose the experimental arm and 12 chose the control arm (Z = -2.000, 0.01 < P = 0.046 < 0.05). 39 male adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 26 males chose the experimental arm and 13 chose the control arm (Z = -2.082, 0.01 < P = 0.037 < 0.05).

[0065] Test 9: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 9 (a hexane solution of 1 μg / μL heptacosanol) for the Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0066] The results showed that 32 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 17 chose the experimental arm and 15 chose the control arm (Z = -0.354, P = 0.724, P > 0.05). 30 male adult *Rhododendron simsii* were also used in the Y-type olfactometer experiment, of which 14 males chose the experimental arm and 16 chose the control arm (Z = -0.362, P = 0.715, P > 0.05).

[0067] Test 10: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 10 (a hexane solution of 2.5 μg / μgL heptadecanol) for a Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0068] The results showed that 31 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 17 chose the experimental arm and 14 chose the control arm (Z = -0.539b, P = 0.590, P > 0.05). 32 male adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 17 males chose the experimental arm and 15 chose the control arm (Z = -0.354, P = 0.724, P > 0.05).

[0069] Test 11: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 11 (a hexane solution of 5 μg / μL heptacosanol) for a Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0070] The results showed that 31 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 21 chose the experimental arm and 10 chose the control arm (Z = -1.976, 0.01 < P = 0.048 < 0.05). 31 male adult *Rhizophora purpurea* were also used in the Y-type olfactometer experiment, of which 19 males chose the experimental arm and 12 chose the control arm (Z = -1.257, P = 0.209, P > 0.05).

[0071] Test 12: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 12 (a hexane solution of 1 μg / μL octadecanaldehyde) for the Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0072] The results showed that 31 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 15 chose the experimental arm and 16 chose the control arm (Z = -0.180, P = 0.857, P > 0.05). 30 male adult *Rhizophora purpurea* were also used in the Y-type olfactometer experiment, of which 16 males chose the experimental arm and 14 chose the control arm (Z = -0.365, P = 0.715, P > 0.05).

[0073] Test 13: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 14 (a hexane solution of 2.5 μg / μL octadecanaldehyde) for the Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0074] The results showed that 32 female adult *Rhododendron simsii* were used in the Y-type olfactometer experiment, of which 15 chose the experimental arm and 17 chose the control arm (Z = -0.354, P = 0.724, P > 0.05). 32 male adult *Rhododendron simsii* were also used in the Y-type olfactometer experiment, of which 17 males chose the experimental arm and 15 chose the control arm (Z = -0.354, P = 0.724, P > 0.05).

[0075] Test 14: The odor source for the experimental arm was 10 μL of the attractant prepared in Example 14 (a hexane solution of 5 μg / μL octadecanaldehyde) for the Y-type olfactometer experiment. The control arm was 10 μL of hexane.

[0076] The results showed that 38 female adult *Rhizophora purpurea* were used in the Y-type olfactometer experiment, of which 18 chose the experimental arm and 20 chose the control arm (Z = -0.324, P = 0.746, P > 0.05). 36 male adult *Rhizophora purpurea* were also used in the Y-type olfactometer experiment, of which 20 males chose the experimental arm and 16 chose the control arm (Z = -0.667, P = 0.505, P > 0.05).

[0077] Based on the above test results Figure 4 The identification results in Table 1, combined with... Figure 5 , Figure 6 The test results show that the attractant for adult *Cephalotaxus fortunei* identified in this invention is a 2.5–5 μg / μL solution of hexane in 2-hexane, which has a good attraction effect on both female and male *Cephalotaxus fortunei*.

[0078] There are currently no reports of the attractant of this invention having an attraction effect on other species of the Cerambycidae family.

[0079] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. Application of attractants in attracting and controlling adult peach red-necked longhorn beetles; The attractant contains hexadecane.

2. The application according to claim 1, characterized in that, The attractant is a hexadecane solution prepared with n-hexane as the solvent.

3. The application according to claim 1 or 2, characterized in that, The concentration of the hexadecane solution in the attractant is 2.5~5 μg / μL.

4. The use of hexadecane in the preparation of attractants for the peach-necked longhorn beetle.

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