Standardized high-throughput screening device and method for fruit tree borer ovicidal agent

Abstract

The application discloses a kind of fruit-tree borer kills ovicidal reagent standardization high-throughput screening device and method, belong to fruit-tree pest reagent screening test equipment technical field, the fruit-tree borer kills ovicidal reagent standardization high-throughput screening device, including cover frame mechanism, base plate mechanism and flexible imitation production egg membrane mechanism, the cover frame mechanism includes cover frame body, a plurality of flow guide ports are integrally formed on the cover frame body.The application is replaced by flexible imitation production egg membrane mechanism as the structural design of real fruit and is used as oviposition carrier, realizes zero damage egg mass acquisition function, and the natural mortality of insect eggs is greatly reduced, effectively improves the accuracy of efficacy evaluation background, by limiting the surface energy value of oviposition membrane body, and the structure layout of independent reaction cabin is formed by cooperating mesh partition and mesh convex strip separation, the function of reagent contact height consistency is realized, the spreading deviation of liquid medicine caused by the surface tension difference of fruit skin is eliminated, the error of egg particle reagent receiving amount is strictly controlled, and the repeatability of test data is improved.

Description

Technical Field

[0001] This invention belongs to the technical field of screening test equipment for fruit tree pests, specifically relating to a standardized high-throughput screening device and method for ovicidal agents against fruit tree fruit borers. Background Technology

[0002] Fruit fruit borer is a major fruit-boring pest that damages pome and stone fruit trees. Once its larvae bore into the fruit, control is extremely difficult. Therefore, the "egg-killing" strategy at the beginning of the pest's life cycle—that is, preventing eggs from hatching or killing newly hatched larvae—is a key link in breaking its chain of damage.

[0003] Currently, the screening of ovicidal agents against codling moths mainly relies on the following two traditional methods, but these methods have limitations: (1) The traditional “egg immersion method” is difficult to operate and has a high damage rate: the eggs of the fruit borer are extremely small (about 0.5-1 mm in diameter) and are flat and oval in shape, closely attached to the surface of the fruit. In order to conduct in vitro testing, the experimenters need to peel the eggs off the fruit peel under a microscope with a dissecting needle or a fine brush. This process is very likely to cause mechanical damage to the eggshell, resulting in a high natural mortality rate in the control group (often exceeding 20%), which seriously interferes with the accurate judgment of the efficacy.

[0004] (2) The "fruit-inoculated egg method" has low throughput and poor consistency: Another commonly used method is to spray pesticides directly onto fruits carrying insect eggs. However, due to individual differences in the thickness of the waxy layer and the density of the trichomes on the fruit surface, the wettability, spreadability, and adhesion of the pesticide solution vary greatly on different fruit surfaces. This interference of surface tension makes the actual deposition amount of the pesticide uneven, which cannot objectively reflect the toxicity of the pesticide itself, and the repeatability of the experimental data is poor.

[0005] (3) Hatching monitoring is cumbersome and inefficient: Traditional hatching observation requires frequent inspection under a stereomicroscope, which is not only labor-intensive, but also easily causes secondary damage to the newly hatched larvae during the sample moving process, making it impossible to achieve continuous and undisturbed monitoring of the egg development process (such as the "blackhead stage").

[0006] Therefore, the pesticide research and development field urgently needs a standardized screening device that can simulate the natural oviposition environment, avoid mechanical damage, ensure consistent pesticide application, and achieve high-throughput, visual monitoring. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide a standardized high-throughput screening device and method for fruit tree fruit moth ovicidal agents.

[0008] The technical solution adopted to solve the above technical problems is: Firstly, a technical solution is provided: a standardized high-throughput screening device for fruit tree fruit borer ovicidal agent, comprising a cover frame mechanism, a substrate mechanism, and a flexible ovicidal membrane mechanism. The cover frame mechanism includes a cover frame body, on which multiple guide ports are integrally formed. The bottom of the cover frame mechanism is provided with a substrate mechanism, which includes a substrate body. The inner side of the substrate body is integrally formed with mesh-like protrusions, and the top of the mesh-like protrusions is integrally formed with an isolation slot. The cover frame mechanism and the substrate mechanism are pressed together to hold a flexible imitation production egg membrane mechanism.

[0009] Furthermore, the guide port is a funnel-shaped structure with a large opening at the top and a small opening at the bottom. Its reverse edge structure not only facilitates the injection of medicine, but also effectively prevents larvae from escaping. Both ends of the cover frame body are integrally formed with a first force strip, which makes it easy for operators to press or separate the cover frame mechanism.

[0010] Through the above technical solution, the funnel-shaped flow guide port integrated into the cover frame body can smoothly complete the quantitative injection of the test reagent. The reverse edge structure of the flow guide port can not only ensure that the liquid flows smoothly into the corresponding independent compartment, but also prevent the hatched larvae from escaping during the cultivation and observation stage, thus improving the airtightness and standardization of the experiment. With the first force strip integrally formed at both ends of the cover frame body, the alignment and pressing between the cover frame mechanism and the base plate mechanism and the disassembly and opening operation can be easily completed, which greatly simplifies the assembly and disassembly process of the device and adapts to the continuous operation requirements of high-throughput batch screening experiments.

[0011] Furthermore, a mesh partition is integrally formed on the inner side of the cover frame body, and a frame cutter is embedded and fixed on the bottom outer edge of the cover frame body for cutting off excess oviposition membrane.

[0012] Through the above technical solution, the mesh partition set inside the cover frame body can be precisely aligned and matched with the internal structure of the substrate mechanism to achieve the regular division of multiple independent test spaces, ensuring that the test of each group of reagents does not interfere with each other. The edge cutter fixed at the bottom outer edge can directly trim and trim the edge of the flexible imitation egg production membrane mechanism that has been laid in place during the pressing and bonding process of the cover frame mechanism, quickly removing excess membrane parts, so that the bonding size of the egg production membrane body is precisely matched with the test area, saving the manual trimming process and improving the assembly efficiency and test regularity of the device.

[0013] Furthermore, at least two magnetically conductive metal sheets are embedded and fixed at the bottom of the cover frame body for attracting the cover frame mechanism and the substrate mechanism.

[0014] Through the above technical solution, the magnetically conductive metal sheet arranged at the bottom of the cover frame body can be precisely magnetically aligned with the corresponding parts of the substrate mechanism, so that the cover frame mechanism and the substrate mechanism are automatically locked and positioned after being pressed together, effectively preventing them from shifting or loosening, ensuring that the flexible imitation production egg membrane mechanism is evenly clamped and fixed. At the same time, the magnetic connection method is convenient to open and close, which not only ensures the overall assembly sealing, but also facilitates the quick disassembly and placement of test pieces after the test, adapting to the operational requirements of batch screening tests.

[0015] Furthermore, the mesh protrusions correspond to the positions of the mesh partitions. When the cover frame mechanism and the base plate mechanism are pressed together, the mesh partitions will be embedded in the isolation slots. At this time, the mesh partitions and mesh protrusions can isolate multiple independent reaction chambers.

[0016] Through the above technical solution, by utilizing the corresponding mesh protrusions and mesh partitions, the mesh partitions are precisely embedded into the isolation slots during the pressing and assembly of the cover frame mechanism and the base plate mechanism, achieving a sealed and fitted positioning of the upper and lower structures. The flexible egg-producing membrane mechanism is sandwiched between the mesh partitions and the isolation slots as a flexible filling pad, which can further improve the sealing and isolation effect between each independent reaction chamber, fill the tiny gaps at the structural fitting points, effectively block the diffusion of water vapor and the seepage of pesticide solution, and thus enclose and separate multiple independent reaction chambers that are not interconnected and have uniform specifications. This completely eliminates the mutual interference of pesticide solution, water vapor and insect eggs in different chambers, effectively avoids the problem of cross-contamination in the experiment, ensures that the environment of each group of pesticide screening experiments is independent and controllable, and greatly improves the experimental accuracy and data repeatability of high-throughput screening of ovicidal pesticides for fruit tree fruit borers.

[0017] Furthermore, the substrate body is made of ultra-clear glass, tempered high-aluminosilicate glass, or scratch-resistant PETG material, which has the characteristics of high transparency and not easy to scratch, making it easy for operators to observe the inside through the substrate body. The bottom of the substrate body is divided into multiple rectangular frames of equal width and length by mesh ridges. A certain amount of sterile water is added to the bottom of each frame to maintain the relative humidity inside the hole at more than 80%, providing the necessary environment for egg incubation.

[0018] Using the above technical solution, the substrate body is made of ultra-clear glass, tempered high-alumina-silica glass, or scratch-resistant PETG material. With its high light transmittance and scratch and wear resistance, it is convenient for testers to directly observe the development status of insect eggs in the chamber in real time through the substrate body. The entire process of microscopic examination and recording can be completed without disassembling the device. The regular rectangular frame space is formed by the mesh convex strips. A certain amount of sterile water is added to the bottom of each area to stably maintain the humidity in the chamber at a standard range of over 80%. This accurately replicates the temperature and humidity environment required for the natural hatching of codling moth eggs, ensuring the normal physiological development of the eggs. This makes the pesticide efficacy screening test more consistent with the actual growth conditions in the field, effectively improving the reference value and authenticity of the test data.

[0019] Furthermore, at least two magnetic absorbing pieces are embedded and fixed on the top of the substrate body, and the positions of the magnetic absorbing pieces correspond to the positions of the magnetically conductive metal pieces. Both ends of the substrate body are integrally formed with second force strips, which facilitates the operator to press or separate the substrate mechanism.

[0020] Through the above technical solution, the magnetic absorbing sheet arranged on the top of the substrate body can be precisely aligned and adsorbed with the magnetically conductive metal sheet at the bottom of the cover frame body, realizing the quick and accurate fitting and locking of the cover frame mechanism and the substrate mechanism. The assembly and positioning are reliable and not easy to misalign. With the second force strip integrally formed at both ends of the substrate body, it can work together with the first force strip to exert force, easily completing the pressing assembly and disassembly of the whole device. The operation is simple and labor-saving, greatly improving the assembly and disassembly efficiency of the test device, and is suitable for continuous operation of large-scale reagent screening tests.

[0021] Furthermore, the flexible oviposition membrane mechanism includes an oviposition membrane body pressed between the cover frame body and the substrate body. The oviposition membrane body is made of low-density polyethylene or polytetrafluoroethylene film. The oviposition membrane body has multiple breathable micropores punctured on it, allowing water vapor at the bottom of the chamber to pass through the breathable micropores to reach the insect eggs.

[0022] The above technical solution securely clamps the oviposition membrane between the cover frame and the substrate, achieving positioning and fixation. The oviposition membrane is made of low-density polyethylene or polytetrafluoroethylene, which is flexible, has good adhesion, and is chemically stable and will not react with the test solution. The breathable micropores on the surface of the oviposition membrane allow for smooth flow of gas and water vapor, enabling the moisture rising from the sterile water on the substrate side to penetrate the membrane and act on the surface of the insect eggs, thus maintaining a stable humidity environment for the insect eggs. This not only meets the humidity requirements for normal egg development but also prevents excessive leakage of the test solution downwards, ensuring a stable and controllable experimental environment.

[0023] Furthermore, the oviposition membrane body is formed with square grid-like or randomly distributed fruit peel-simulating grooves with a depth of 0.5-1mm through a micro-embossing process to simulate the rough texture of the fruit peel.

[0024] Through the above technical solution, a square grid pattern or random concave pattern with a depth of 0.5-1mm is formed on the surface of the oviposition membrane using a micro-embossing process. This can highly replicate the rough texture and feel of the fruit skin of fruit trees, conforming to the natural oviposition and attachment habits of adult fruit borers. This can effectively enhance the adults' willingness to actively lay eggs and the stability of egg attachment, ensuring that the attachment state and distribution of eggs are consistent with the natural oviposition state in the field. This eliminates the deviation in oviposition behavior caused by differences in substrate materials, making the results of subsequent ovicidal pesticide screening tests closer to the actual application effect in the field, and improving the simulation and practical reference of the test data.

[0025] Secondly, based on the first aspect mentioned above, a method for a standardized high-throughput screening device for fruit tree fruit borer ovicidal agents is also provided, comprising the following specific steps: Step 1: Standardized oviposition induction: The oviposition membrane is flatly attached to the inner wall of the oviposition cage, and newly emerged adult codling moths (male-to-female ratio 1:1) are introduced. Under dark conditions (photoperiod L:D=14:10), female moths are induced to lay eggs on the membrane for 24 hours. Step 2: Egg mass screening and loading: Take out the oviposition membrane body and place it under a stereomicroscope; screen out membranes with moderate egg density, uniform egg attachment, and no overlapping or pressing; lay this membrane (egg side up) flat on the base plate body and fix it with the cover frame mechanism; Step 3: High-throughput precision application: Using 12 or 8 spray guns, 5-20 μL of the test solution of a series of concentrations is precisely dripped or sprayed into each independent reaction chamber through the guide port of the cover frame body. The solution is evenly covered on the egg mass on the surface of the oviposition membrane body under the action of gravity. Step 4: Humidity incubation and dynamic monitoring: Place the assembled device in an artificial intelligence climate incubator (temperature 25±1℃, humidity 85±5%). Since the substrate is made of highly transparent material, the operator can directly place the device on the stage of an inverted microscope and continuously image the eggs in the same field of view every day without damaging the device's seal, recording their development process (from milky white → light red → blackhead). Step 5: Efficacy evaluation: Count the number of unhatched and colorless dead eggs at 24 hours and 48 hours after application to calculate the ovicidal rate.

[0026] The beneficial effects of this invention are as follows: 1. This invention achieves zero-damage egg-laying mass acquisition by replacing real fruit with a flexible oviposition membrane structure. Adult insects can directly lay eggs on the detachable oviposition membrane, eliminating the need for egg removal. This significantly reduces the natural mortality rate of insect eggs in the control group and effectively improves the accuracy of the baseline for drug efficacy evaluation.

[0027] 2. This invention achieves a consistent contact height with the agent by limiting the surface energy value of the oviposition membrane and using a structural layout that separates the reaction chambers with mesh partitions and mesh ridges. This eliminates the deviation in agent spreading caused by differences in the surface tension of the fruit peel, strictly controls the error in the amount of agent received by the eggs, and significantly improves the repeatability of experimental data.

[0028] 3. This invention uses a combination structure of a substrate body made of highly transparent material and a light-transmitting oviposition membrane body with breathable micropores to achieve the function of visual monitoring of the entire development cycle of insect eggs. Without moving or touching the egg mass, the entire development process of insect eggs can be tracked with the help of microscopic equipment, and the effects of fast-acting and slow-acting agents can be accurately distinguished.

[0029] 4. This invention uses a cover frame mechanism to arrange multiple sets of guide ports and an integrated structure that adapts to the standard orifice plate specifications to achieve high-throughput operation of reagent screening. It can be used with a multi-stage gun to complete batch and precise sample addition operations, greatly reducing test time and multiplying the overall work efficiency of reagent screening. Attached Figure Description

[0030] Figure 1 This is a three-dimensional structural diagram of the present invention; Figure 2 This is a three-dimensional sectional view of the present invention; Figure 3 This is a first-view schematic diagram of the cover frame mechanism of the present invention; Figure 4 This is a second-view schematic diagram of the cover frame mechanism of the present invention; Figure 5 This is a schematic diagram of the substrate mechanism of the present invention; Figure 6 yes Figure 2 Enlarged view of point A in the middle; Figure 7 yes Figure 2 Enlarged view of point B in the middle; Figure 8 This is a flowchart of the operation method of the present invention.

[0031] Reference numerals: 1. Cover frame mechanism; 101. Cover frame body; 102. Flow guide; 103. First force application strip; 104. Mesh partition; 105. Frame cutter; 106. Magnetic conductive metal sheet; 2. Substrate mechanism; 201. Substrate body; 202. Mesh protrusion; 203. Isolation slot; 204. Sterile water; 205. Magnetic suction piece; 206. Second force application strip; 3. Flexible simulated egg-laying membrane mechanism; 301. Egg-laying membrane body; 302. Breathable micropores; 303. Fruit peel simulated concave texture. Detailed Implementation

[0032] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0033] like Figures 1-8As shown, a standardized high-throughput screening device for fruit tree fruit borer ovicidal agent includes a cover frame mechanism 1, a substrate mechanism 2, and a flexible oviposition membrane mechanism 3. The cover frame mechanism 1 includes a cover frame body 101, and the guide port 102 is a funnel-shaped structure with a large opening at the top and a small opening at the bottom. Its reverse edge structure not only facilitates the injection of the pesticide solution but also effectively prevents larvae from escaping. Both ends of the cover frame body 101 are integrally formed with first force strips 103, which facilitates the operator to press or separate the cover frame mechanism 1. The inner side of the cover frame body 101 is integrally formed with a mesh partition 104, and the bottom end of the cover frame body 101... A frame cutter 105 is inlaid and fixed on the outer edge for cutting off excess oviposition membrane. At least two magnetically conductive metal sheets 106 are inlaid and fixed on the bottom of the cover frame body 101 for attracting the cover frame mechanism 1 and the base plate mechanism 2. The funnel-shaped guide port 102 integrally arranged on the cover frame body 101 can smoothly complete the quantitative injection of the test reagent. The reverse edge structure of the guide port 102 can ensure that the liquid can flow smoothly into the corresponding independent compartment, and can also prevent the hatched larvae from escaping during the cultivation and observation stage, thereby improving the airtightness and standardization of the experiment. Combined with the first force strip 103 integrally formed at both ends of the cover frame body 101, The alignment and pressing of the cover frame mechanism 1 and the substrate mechanism 2 can be easily completed, as well as the disassembly and opening operations, greatly simplifying the device assembly and disassembly process. It is suitable for the continuous operation requirements of high-throughput batch screening tests. The mesh partition 104 set on the inner side of the cover frame body 101 can be precisely aligned and matched with the internal structure of the substrate mechanism 2 to realize the regular division of multiple independent test spaces and ensure that the reagent tests of each group do not interfere with each other. The edge cutter 105 fixed on the bottom outer edge can directly trim and trim the edge of the flexible imitation production egg membrane mechanism 3 that has been laid in place during the pressing and bonding process of the cover frame mechanism 1, and quickly remove excess membrane parts. The design allows the egg-laying membrane body 301 to precisely match the test area, eliminating the need for manual trimming and improving the assembly efficiency and test regularity of the device. The magnetically conductive metal sheet 106 at the bottom of the cover frame body 101 can precisely magnetically attach to the corresponding parts of the substrate mechanism 2, so that the cover frame mechanism 1 and the substrate mechanism 2 are automatically locked and positioned after being pressed together, effectively preventing them from shifting or loosening. This ensures that the flexible imitation egg-laying membrane mechanism 3 is evenly clamped and fixed. At the same time, the magnetic connection method is convenient to open and close, ensuring the overall assembly is sealed and facilitating the quick disassembly and placement of test pieces after the test, which is suitable for the operation requirements of batch screening tests.

[0034] like Figure 2 and Figure 4As shown, a substrate mechanism 2 is provided at the bottom of the cover frame mechanism 1. The substrate mechanism 2 includes a substrate body 201. A mesh-like protrusion 202 is integrally formed on the inner side of the substrate body 201, and an isolation slot 203 is integrally formed on the top of the mesh-like protrusion 202. The mesh-like protrusion 202 corresponds to the position of the mesh partition 104. When the cover frame mechanism 1 and the substrate mechanism 2 are pressed together, the mesh partition 104 will be embedded in the isolation slot 203. At this time, the mesh partition 104 and the mesh protrusion 202 can isolate multiple independent reaction chambers. The substrate body 201 is made of ultra-clear glass, tempered high-alumina-silicon glass, or scratch-resistant PETG material, which has the characteristics of high transparency and scratch resistance, making it easy for operators to see through the substrate body 201. 1. Observing the interior, the bottom of the substrate body 201 is divided into multiple rectangular frames of equal width and length by mesh protrusions 202. A certain amount of sterile water 204 is added to the bottom of each frame to maintain the relative humidity inside the hole at more than 80%, providing the necessary environment for egg incubation. At least two magnetic absorbing pieces 205 are embedded and fixed on the top of the substrate body 201. The positions of the magnetic absorbing pieces 205 correspond to the positions of the magnetic conductive metal pieces 106. The two ends of the substrate body 201 are integrally formed with second force strips 206, which can facilitate the operator to press or separate the substrate mechanism 2. Using the mesh protrusions 202 and mesh partitions 104 that correspond to each other, when the cover frame mechanism 1 and the substrate mechanism 2 are pressed and assembled, the mesh... The mesh partition 104 is precisely embedded inside the isolation slot 203, achieving a sealed and fitted positioning of the upper and lower structures. A flexible, egg-producing membrane mechanism 3 is sandwiched between the mesh partition 104 and the isolation slot 203, acting as a flexible filling layer. This further enhances the sealing and isolation effect between the independent reaction chambers, filling the tiny gaps at the structural fitting points and effectively blocking moisture diffusion and pesticide seepage. This effectively separates multiple independent reaction chambers that are not interconnected and have uniform specifications, completely eliminating cross-contamination of pesticides, moisture, and insect eggs between different chambers. This effectively avoids cross-contamination during testing, ensuring that the testing environment for each pesticide screening experiment is independent and controllable, significantly improving the accuracy and quantity of high-throughput screening of ovicidal pesticides for fruit tree fruit borers. For repeatability, the substrate body 201 is made of ultra-clear glass, tempered high-alumina-silica glass, or scratch-resistant PETG material. Its high light transmittance and scratch / wear resistance allow researchers to directly observe the development of insect eggs inside the chamber in real time without disassembling the device. A regular rectangular space is formed by mesh-like raised strips 202. Adding a measured amount of sterile water 204 to the bottom of each area maintains a stable humidity level above 80%, accurately replicating the temperature and humidity environment required for the natural hatching of codling moth eggs. This ensures normal physiological development of the eggs, making pesticide efficacy screening experiments more closely resemble actual field growth conditions and effectively improving the reference value and authenticity of the experimental data.The magnetic absorbing sheet 205 arranged on the top of the substrate body 201 can be precisely aligned and attracted with the magnetically conductive metal sheet 106 at the bottom of the cover frame body 101, realizing quick and precise bonding and locking of the cover frame mechanism 1 and the substrate mechanism 2. The assembly and positioning are reliable and not prone to misalignment. In conjunction with the second force strip 206 integrally formed at both ends of the substrate body 201, it can work together with the first force strip 103 to easily complete the pressing assembly and disassembly of the entire device. The operation is simple and labor-saving, greatly improving the assembly and disassembly efficiency of the test device, and is suitable for continuous operation of large-scale reagent screening tests.

[0035] like Figure 2 and Figure 5 As shown, a flexible simulated egg-laying membrane mechanism 3 is clamped between the cover frame mechanism 1 and the base plate mechanism 2. The flexible simulated egg-laying membrane mechanism 3 includes an egg-laying membrane body 301 clamped between the cover frame body 101 and the base plate body 201. The egg-laying membrane body 301 is made of low-density polyethylene or polytetrafluoroethylene film. Multiple permeable micropores 302 are needle-punched on the egg-laying membrane body 301, allowing moisture from the bottom of the chamber to pass through the micropores 302 to reach the eggs. The egg-laying membrane body 301 has square grid-like or randomly distributed fruit peel-simulating grooves 303 with a depth of 0.5-1mm formed by a micro-embossing process to simulate the rough texture of the fruit peel. The egg-laying membrane body 301 is securely clamped between the cover frame body 101 and the base plate body 201 for positioning and fixation. The egg-laying membrane body 301 is made of low-density polyethylene or polytetrafluoroethylene, a material that is flexible, has good adhesion, and is chemically stable and will not react with the test solution. The breathable micropores 302 on the surface of the membrane body 301 allow for smooth flow of gas and water vapor, enabling the moisture rising from the sterile water 204 on the substrate side to penetrate the membrane and act on the surface of the insect eggs. This stabilizes the humidity of the environment where the insect eggs are located, meeting the humidity requirements for normal egg development while preventing excessive leakage of pesticide solution downwards, ensuring a stable and controllable experimental environment. The surface of the oviposition membrane body 301 is processed using a micro-embossing process to form a square grid pattern or random concave pattern with a depth of 0.5-1mm. This highly replicates the rough texture and feel of the fruit skin of fruit trees, conforming to the natural oviposition and attachment habits of adult fruit borers. This effectively enhances the adults' willingness to actively lay eggs and the stability of egg attachment, ensuring that the attachment state and distribution of the eggs are consistent with the natural oviposition state in the field. This eliminates the deviation in oviposition behavior caused by differences in substrate materials, making the results of subsequent ovicidal pesticide screening tests closer to the actual application effect in the field, and improving the simulation and practical reference of the experimental data.

[0036] A method for a standardized high-throughput screening device for fruit tree fruit borer ovicidal agents includes the following specific steps: Step 1: Standardized oviposition induction: The oviposition membrane body 301 is flatly attached to the inner wall of the oviposition cage, and adult codling moths (female to male ratio 1:1) are introduced. Under dark conditions (photoperiod L:D=14:10), female moths are induced to lay eggs on the membrane for 24 hours. Step 2: Egg mass screening and loading: Take out the oviposition membrane body 301 and place it under a stereomicroscope; screen out membranes with moderate egg density, uniform egg attachment, and no overlapping or pressing; lay this membrane (egg side up) flat on the substrate body 201 and fix it with the cover frame mechanism 1; Step 3: High-throughput precision application: Using 12 or 8 spray guns, through the guide port 102 of the cover frame body 101, precisely drip or spray 5-20μL of the test solution of a series of concentrations into each independent reaction chamber. Under the action of gravity, the solution is evenly covered on the egg mass on the surface of the egg-laying membrane body 301. Step 4: Constant humidity culture and dynamic monitoring: Place the assembled device in an artificial intelligence climate incubator (temperature 25±1℃, humidity 85±5%). Since the substrate body 201 is made of highly transparent material, the operator can directly place the device on the stage of an inverted microscope and continuously image the eggs in the same field of view every day without damaging the device's seal, recording their development process (from milky white → light red → blackhead). Step 5: Efficacy evaluation: Count the number of unhatched and colorless dead eggs at 24 hours and 48 hours after application to calculate the ovicidal rate.

[0037] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention.

Claims

1. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers, characterized in that: The system includes a cover frame mechanism (1), a substrate mechanism (2), and a flexible imitation production egg membrane mechanism (3). The cover frame mechanism (1) includes a cover frame body (101), on which multiple flow guides (102) are integrally formed. The bottom of the cover frame mechanism (1) is provided with a substrate mechanism (2), which includes a substrate body (201). The inner side of the substrate body (201) is integrally formed with a mesh protrusion (202), and the top of the mesh protrusion (202) is integrally formed with an isolation slot (203). The flexible imitation production egg membrane mechanism (3) is held in a pressing clamp between the cover frame mechanism (1) and the substrate mechanism (2).

2. The standardized high-throughput screening device for fruit tree fruit borer ovicidal agents according to claim 1, characterized in that, The guide port (102) is a funnel-shaped structure with a large opening at the top and a small opening at the bottom. Its reverse edge structure not only facilitates the injection of medicine, but also effectively prevents larvae from escaping. Both ends of the cover frame body (101) are integrally formed with a first force strip (103), which makes it easy for operators to press or separate the cover frame mechanism (1).

3. The standardized high-throughput screening device for fruit tree fruit borer ovicidal agents according to claim 1, characterized in that, The inner side of the cover frame body (101) is integrally formed with a mesh partition (104), and the bottom outer edge of the cover frame body (101) is inlaid with a frame cutter (105) for cutting off excess oviposition membrane.

4. The standardized high-throughput screening device for fruit tree fruit borer ovicidal agents according to claim 1, characterized in that, At least two magnetically conductive metal sheets (106) are embedded and fixed at the bottom of the cover frame body (101) for attracting the cover frame mechanism (1) and the base plate mechanism (2).

5. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers according to claim 3, characterized in that, The mesh protrusions (202) correspond to the mesh partitions (104). When the cover frame mechanism (1) and the base plate mechanism (2) are pressed together, the mesh partitions (104) will be embedded in the isolation slots (203). At this time, the mesh partitions (104) and the mesh protrusions (202) can isolate multiple independent reaction chambers.

6. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers according to claim 1, characterized in that, The substrate body (201) is made of ultra-white glass, tempered high-aluminosilicate glass or scratch-resistant PETG material, which has the characteristics of high transparency and not easy to scratch, making it easy for operators to observe the inside through the substrate body (201). The bottom of the substrate body (201) is divided into multiple rectangular frames of equal width and length by mesh protrusions (202). A certain amount of sterile water (204) is added to the bottom of each frame to maintain the relative humidity inside the hole at more than 80%, providing the necessary environment for egg incubation.

7. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers according to claim 4, characterized in that, At least two magnetic absorbing pieces (205) are embedded and fixed on the top of the substrate body (201). The position of the magnetic absorbing pieces (205) corresponds to the position of the magnetic conductive metal piece (106). Both ends of the substrate body (201) are integrally formed with a second force strip (206), which makes it easy for the operator to press or separate the substrate mechanism (2).

8. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers according to claim 1, characterized in that, The flexible imitation egg-laying membrane mechanism (3) includes an egg-laying membrane body (301) pressed between the cover frame body (101) and the substrate body (201). The egg-laying membrane body (301) is made of low-density polyethylene or polytetrafluoroethylene film. The egg-laying membrane body (301) is needled with multiple breathable micropores (302), and water vapor at the bottom of the chamber can pass through the breathable micropores (302) to reach the insect eggs.

9. A standardized high-throughput screening device for ovicidal agents against fruit tree fruit borers according to claim 8, characterized in that, The egg-laying membrane body (301) is formed with a micro-embossing process to create square grid-like or randomly distributed fruit peel-simulating grooves (303) with a depth of 0.5-1mm, in order to simulate the rough touch of the fruit peel.

10. A method of using a standardized high-throughput screening device for fruit tree fruit borer ovicidal agents, wherein the device is applied to the standardized high-throughput screening device for fruit tree fruit borer ovicidal agents according to any one of claims 1-9, characterized in that, The specific steps include the following: Step 1: Standardized oviposition induction: The oviposition membrane body (301) is flatly attached to the inner wall of the oviposition cage, and adult codling moths (male-to-female ratio 1:1) are introduced. Under dark conditions (photoperiod L:D=14:10), female moths are induced to lay eggs on the membrane for 24 hours. Step 2: Egg mass screening and loading: Take out the oviposition membrane body (301) and place it under a stereomicroscope; screen out membranes with moderate egg density, uniform egg attachment, and no overlapping or pressing; lay this membrane (egg side up) flat on the substrate body (201) and fix it with the cover frame mechanism (1); Step 3: High-throughput precision application: Using 12 or 8 spray guns, through the guide port (102) of the cover frame body (101), precisely drip or spray 5-20 μL of the test solution of a series concentration into each independent reaction chamber. The solution is evenly covered on the egg mass on the surface of the egg-laying membrane body (301) under the action of gravity. Step 4: Humidity incubation and dynamic monitoring: Place the assembled device in an artificial intelligence climate incubator (temperature 25±1℃, humidity 85±5%). Since the substrate (201) is made of highly transparent material, the operator can directly place the device on the stage of an inverted microscope and continuously image the eggs in the same field of view every day without damaging the device's seal, recording their development process (from milky white → light red → blackhead). Step 5: Efficacy evaluation: Count the number of unhatched and colorless dead eggs at 24 hours and 48 hours after application to calculate the ovicidal rate.

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