Apparatus and method for determining behavioral laterality in insect antennae

Abstract

The application belongs to the technical field of insect electrophysiological behavior, and discloses a device and method for measuring the behavioral lateralization of insect antennae. The device comprises a telescopic multi-section sleeve fixed at one end to an antenna potential instrument through a bayonet, and the other end of the telescopic multi-section sleeve is connected to a microtube, which is used to cover a single antenna of the measured insect and prevent the measured insect from being affected by sample airflow. The diameter of the microtube is greater than the diameter of the thickest part of the antenna of the measured insect, and the length of the microtube is greater than the length of the antenna of the insect. The method for measuring the behavior of the left and right antennae of the insect based on the device is simple and convenient, can be used for long-time measurement of various compound components of different types, can effectively solve the problem that the behavioral differences between the left and right antennae of the insect are ignored in actual tests, can enhance the signal substances ignored due to unobvious reactions, and is helpful for perfecting or even changing the composition of insect pheromones.

Description

Technical Field

[0001] The present invention belongs to the field of insect electrophysiological behavior, and more particularly relates to a device and method for measuring the laterality of insect antennae behavior. Background Art

[0002] For most insects, behavioral laterality is investigated through the study of left-right antennae asymmetry. As a crucial component of the insect sensory system, antennae play a crucial role in social behaviors such as host location, recognition, feeding, mate foraging, mating, reproduction, roosting, defense, migration, and flight rate stabilization (Ochieng et al., 2000; Lu et al., 2007; Fuller et al., 2014). Antennae are also a crucial signal carrier for identifying insect pheromones and screening for plant-derived odors. Insect antennae typically exist in pairs. Our preliminary research has revealed that left and right antennae differ in host selection, and that the left and right antennae also differ in their selection of pheromones between sexes.

[0003] Currently, laterality in insect behavior and function is a crucial area of ​​research, providing a deeper understanding of the early origins of laterality in biodiversity and holding significant scientific significance for exploring insect evolution (Anderson et al., 2021). Reports on behavioral laterality have been published in Hymenoptera, Diptera, Hemiptera, Blattodea, and Orthoptera, but these studies have focused on brain functional laterality and asymmetry in gene expression, with no reports on laterality in electrophysiological behavior. Antennae are widely used in nature to screen for active compounds. During electroantennal potential (EPA) assays, antennal activity decreases over time, with activity in isolated antennae typically lasting less than an hour. Therefore, ideally, live insects would be ideally suited for this purpose. However, this approach is extremely complex and difficult to implement. In the identification of insect sex pheromones using gas chromatography-electroantennal potential (GC-EAD), differences between left and right antennae are often overlooked, and random antennae are used for testing, exacerbating response variations between individuals. By measuring the difference in left and right antennae and thus determining the sensitivity of left and right antennae, it is possible to increase the number of signal substances that may have been overlooked due to their less obvious responses, helping to improve or even modify the composition of insect pheromones. Therefore, the present invention provides a device for isolating left and right insect antennae, and uses this device for the first time to measure the electrophysiological behavioral differences between the left and right antennae of insects, providing important research basis for the identification of trace signal substances in insects and the field of evolution.

[0004] Therefore, how to provide a device and method for measuring the laterality of insect antennae behavior is an urgent problem that needs to be solved by those skilled in the art. Summary of the Invention

[0005] To overcome the shortcomings and deficiencies of the prior art, the present invention provides a device and method for measuring the laterality of insect antennal behavior. This isolation device can completely isolate the antennae of a live insect, preventing them from being affected by other odors. The method for measuring the behavior of the left and right antennae of an insect using this device is simple and convenient, capable of performing long-term measurements on a variety of different compound components. It effectively addresses the problem of overlooked differences in insect antennal behavior in actual experiments, enhances signal substances that are often overlooked due to their lack of significant responses, and helps improve or even modify the composition of insect pheromones. For example, in a gas chromatography-electroantennary (GC-EAD) experiment verifying the composition of the diamondback moth sex pheromone, when the sex pheromone content in the gonads of female P. diamondback moths is low, the right antennae show very little, if any, reaction to the trace component cis-11-hexadecenol. However, when the left antennae are used for the experiment, a certain response value to the trace component cis-11-hexadecenol is detected, indicating a certain signal. Numerous research papers and field trials have shown that while cis-11-hexadecenol is a minor component of the diamondback moth sex pheromone, it significantly enhances the main component, enhancing the trapping of diamondback moth in the field. Therefore, identifying minor components of sex pheromones is crucial. Therefore, the device of the present invention can screen for more useful active compounds when detecting unknown pheromones or plant-derived volatiles from certain insects.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A device for measuring the laterality of insect antennae's behavior, comprising a retractable multi-section sleeve with one end fixed to an antennae potentiometer via a bayonet, the other end of the retractable multi-section sleeve being connected to a microtube, the microtube being used to sheath a single antennae of the insect being measured to prevent it from being affected by the sample airflow.

[0008] Preferably, the diameter of the microtube is larger than the diameter of the thickest part of the antennae of the insect being measured, and the length of the microtube is larger than the length of the antennae of the insect.

[0009] The beneficial effect of the above technical solution is that the microtubes can isolate the mutual interference of odors on the two antennae, and when one antennae is measured, the other antennae are not affected.

[0010] Preferably, the material of the microtube is selected from one of glass, fiber, latex, polyethylene, and polytetrafluoroethylene.

[0011] Preferably, the material of the telescopic multi-section sleeve is selected from one of silver, copper and aluminum.

[0012] The beneficial effects of the above technical solution are: the sleeve has good toughness and is retractable and fixable.

[0013] The present invention also provides a method for measuring the laterality of insect antennae behavior, using the above-mentioned device, comprising the following steps:

[0014] (1) The antennae are the left and right antennae of the same insect. A pipette tip is selected, the bottom portion is removed, and a whole live insect is used. The insect is fixed in the tip, leaving only the head exposed. The pipette tip is fixed using the micromanipulator in the potentioantenometer.

[0015] (2) Use a microtube to cover the left or right antennae of the insect, connect the positive end of the glass electrode containing conductive liquid in the antennae potentiometer to the top of the left or right antennae of the insect, and insert the negative end of the glass electrode containing conductive liquid into the base of the antennae to be measured. The direction of the sample airflow is the middle and upper part of the antennae to be measured.

[0016] It can be seen from the above technical solution that, compared with the prior art, the present invention provides a device and method for measuring the laterality of insect antennae behavior, which has the following beneficial effects:

[0017] (1) The device of the present invention is simple and convenient to use. The device is fixed on the potentioantenometer, has a certain degree of elasticity, provides good protection for non-test antennae, and can effectively isolate the interference of external odors.

[0018] (2) The assay method based on the device of the present invention is of great significance, as it can be used to measure a variety of different types of compound components over a long period of time. Conventionally, antennal potential tests are performed using excised antennae (i.e., cut antennae). This results in decreased antennal activity, and the entire test can only be completed in a short period of time. Otherwise, the antennae will lose activity, making it impossible to test multiple samples from the same batch at once.

[0019] (3) In existing practical experiments, the behavioral differences between the left and right antennae of insects are often overlooked, and the experiments have a certain degree of randomness. The present invention can utilize the differences in the electrophysiological behavior of the left and right antennae to effectively enhance signal substances that are often overlooked due to their lack of obvious reactions, helping to improve or even change the composition of insect pheromones, and has important significance for the comprehensive application of green pest control technologies. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0021] Figure 1Schematic diagram of a device for measuring the laterality of insect antennae behavior according to the present invention.

[0022] Figure 2 Schematic diagram of using the device of the present invention to measure antenna potential.

[0023] Figure 3 This is the first measurement of the right antennal potential of the diamondback moth in Example 1 of the present invention.

[0024] Figure 4 This is the first measurement of the left antennae potential of the diamondback moth in Example 1 of the present invention.

[0025] Figure 5 The right antennal potential of the diamondback moth was measured again in Example 1 of the present invention.

[0026] Figure 6 The left antennae potential of the diamondback moth was measured again in Example 1 of the present invention.

[0027] Figure 7 This is the first measurement of the right antennal potential of Plutella xylostella in Comparative Example 1 of the present invention.

[0028] Figure 8 This is the first measurement of the left antennae potential of Plutella xylostella in Comparative Example 1 of the present invention.

[0029] Figure 9 The right antennal potential of the diamondback moth was measured again for Comparative Example 1 of the present invention.

[0030] Figure 10 The left antennae potential of the diamondback moth was measured again for Comparative Example 1 of the present invention.

[0031] Among them: 1- gun tip, 2- micropipette, 3- retractable multi-section cannula, 4- bayonet, 5- positive electrode, 6- negative electrode, 7- insect antennae, 8- air flow purge port. DETAILED DESCRIPTION

[0032] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0033] Example 1

[0034] A device for measuring the laterality of insect antennae's behavior, comprising a retractable multi-section sleeve with one end fixed to an antennae potentiometer via a bayonet, the other end of the retractable multi-section sleeve being connected to a microtube, the microtube being used to sheath a single antennae of the insect being measured to prevent it from being affected by the sample airflow.

[0035] The diameter of the microtubule is greater than the diameter of the thickest part of the antennae of the measured insect, and the length of the microtubule is greater than the length of the antennae of the insect.

[0036] The material of the microtube is fiber.

[0037] The material of the telescopic multi-section sleeve is copper.

[0038] A method for determining the laterality of insect antennal behavior uses a whole live diamondback moth insect. The diamondback moth is a second-instar adult after eclosion, eclosing alone in a finger-shaped tube, and has not mated before the experiment. First, a suitable pipette tip is selected, the bottom is subtracted, and the insect to be measured is fixed in the tip, leaving only the head exposed. The pipette tip is fixed using the micromanipulator in the antennae potentiometer, and the distance between the insect antennae and the two electrodes of the potentiometer is slowly adjusted. When the distance is appropriate, the micropipette is placed around the left or right antennae of the insect. The positive end of the glass electrode containing conductive liquid in the antennae potentiometer is connected to the top of the left or right antennae of the insect, and the negative end of the glass electrode containing conductive liquid is inserted into the base of the antennae to be measured. The sample solution to be tested is the plant-derived volatile cis-3-hexenol, diluted to 10 with liquid paraffin. -2 The concentration (volume ratio) is evenly dropped on a "V"-shaped filter paper strip with a length of 5 cm and a width of 0.5 cm, and placed in a Pasteur pipette. The end of the pipette is connected to the stimulating airflow control device. The air supply tube mouth is perpendicular to the longitudinal direction of the antennae and is 1 cm away from the antennae. The airflow sweep direction is the upper and middle part of the antennae to be measured. Adjust the continuous gas flow rate to 124 ml / min, the stimulating gas flow rate to 20 ml / min, the stimulation time for each time is 0.2 s, and the interval between two stimulations is 40 s. The retractable multi-section sleeve is connected to the contact angle potentiostat through the bayonet. After the connection is completed, the laterality of the electrophysiological behavior of the left and right antennae of the insect is detected. 2 hours after the first measurement, the measurement is carried out again using the same method.

[0039] Comparative Example 1

[0040] Electrophysiological behavior of isolated insect antennae was measured using a potentioantennometer. Diamondback moths were anesthetized with carbon dioxide, and then either the left or right antenna was removed for testing. After the test was complete, the other antenna was removed for testing. Two hours after the initial measurement, the test was repeated using the same method.

[0041] from Figure 3-Figure 6 It can be seen that when the insect left / right antennae isolation device of the present invention is used to protect the left / right antennae from interference by the sample airflow and maintain the antennae's persistent activity, the electrophysiological response values ​​of the left and right antennae of the diamondback moth can be measured well, and the electrophysiological behavior differences of the left and right antennae can be compared. The antennae always maintain a stable and high activity. The electrophysiological response value of the left antennae is higher than that of the right antennae ( Figure 3 and Figure 4 , Figure 5 and Figure 6), which will allow the test substance to be detected with a certain reaction intensity even at extremely low concentrations, and ensure that the activity of certain important compounds is not easily overlooked by errors caused by random testing of the left and right antennae.

[0042] Figure 7-10 The electrophysiological response potentials of the left and right antennae of the isolated diamondback moth were measured. Compared to the electrophysiological response values ​​of the whole live insect, the antennae's electrophysiological response values ​​were smaller, and the peak shape was sometimes regular and sometimes irregular, with a double peak. When the test was repeated two hours after the experiment, the activity of the isolated antennae had greatly decreased, and the response intensity was relatively low in both the left and right antennae ( Figure 9 and Figure 10 ).

[0043] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. Reference can be made to the common and similar parts between the various embodiments. For the solutions disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple, and the relevant parts can be referred to the method section.

[0044] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A device for measuring the laterality of insect antennae behavior, characterized in that: The device includes a retractable multi-section sleeve with one end fixed to the antenna potentiometer through a bayonet, and the other end of the retractable multi-section sleeve is connected to a microtube, which is used to sheath a single antenna of the measured insect to prevent it from being affected by the sample airflow.

2. The device for measuring the laterality of insect antennae behavior according to claim 1, characterized in that: The diameter of the microtubule is greater than the diameter of the thickest part of the antennae of the measured insect, and the length of the microtubule is greater than the length of the antennae of the insect.

3. The device for measuring the laterality of insect antennae behavior according to claim 1, characterized in that: The material of the microtube is selected from one of glass, fiber, latex, polyethylene and polytetrafluoroethylene.

4. The device for measuring the laterality of insect antennae behavior according to claim 1, characterized in that: The material of the telescopic multi-section sleeve is selected from one of silver, copper and aluminum.

5. A method for determining the laterality of insect antennae behavior, characterized in that: The device according to any one of claims 1 to 4 comprises the following steps: (1) The antennae are the left and right antennae of the same insect. A pipette tip is selected, the bottom portion is removed, and a whole live insect is used. The insect is fixed in the tip, leaving only the head exposed. The pipette tip is fixed using the micromanipulator in the potentioantenometer. (2) Use a microtube to cover the left or right antennae of the insect, connect the positive end of the glass electrode containing conductive liquid in the antennae potentiometer to the top of the left or right antennae of the insect, and insert the negative end of the glass electrode containing conductive liquid into the base of the antennae to be measured. The direction of the sample airflow is the middle and upper part of the antennae to be measured.

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