An ecdysone-binding protein-derived peptide and biosensor capable of effectively monitoring the sex pheromone of the fall armyworm

Abstract

The application discloses an information pheromone binding protein derivative peptide and a biosensor capable of effectively monitoring the sex pheromone of Hyphantria cunea, and belongs to the field of biosensors. At present, the monitoring of H. cunea is usually achieved by means of sample plot investigation, sex lures and other means, and problems such as poor timeliness, high labor cost and insufficient data continuity exist. In order to solve the above problems, the information pheromone binding protein derivative peptide provided by the application has an amino acid sequence as shown in SEQ ID NO: 2, the biosensor comprises a substrate and an interdigital electrode, single-walled carbon nanotubes are attached to the electrode, and the information pheromone binding protein derivative peptide is further connected. The sensor can specifically detect the sex pheromone of H. cunea and has no response to 17 kinds of plant volatiles, and can detect the sex pheromone released by as low as five live female H. cunea under laboratory conditions. The sensor has application potential in the fields of early-stage pest monitoring, improvement of prevention and control efficiency and guarantee of ecological safety.

Description

TECHNICAL FIELD

[0001] The present application relates to an information pheromone binding protein derived peptide and biosensor capable of effectively monitoring the sex pheromone of Hyphantria cunea, and belongs to the field of biosensors. BACKGROUND

[0002] Hyphantria cunea is one of the most notorious pests in global agriculture and forestry. As an omnivorous pest, it feeds on more than 300 species of trees and crops such as poplar, willow, elm, mulberry, apple, corn, and soybean. A single female Hyphantria cunea can lay 500-1500 eggs, and it has 2-3 generations per year. The high reproductive rate is another reason why it is a major threat to the agro-ecosystem. Therefore, the current priority is to take sustained and effective measures to curb the harm of this pest. Early detection and dynamic monitoring of the population is the key to curb the outbreak of the pest. In particular, accurate detection and understanding of the occurrence regularity and population trend of Hyphantria cunea are important prerequisites for scientific prevention and control. The current commonly used monitoring methods, such as plot investigation, field sampling, and sex lure trap monitoring, have problems such as poor timeliness, high labor cost, and insufficient data continuity, which makes it difficult to meet the needs of rapid response for prevention and control. It is of great significance to develop low-cost, high-sensitivity, real-time monitoring technology and build a multi-source data fusion implementation monitoring system to improve the efficiency of Hyphantria cunea prevention and control and protect ecological safety. It is expected to provide an innovative technical path for pest integrated management.

[0003] Biosensors have been widely used for gas detection. Metal, metal oxide and carbon-based materials have been used as sensing materials for detecting various gases. However, the selectivity of these traditional materials is usually insufficient. In recent years, odorant binding protein (OBP) and odorant binding protein-derived peptide (OBPP) are emerging as a new type of gas detection sensing material. Odorant binding protein is responsible for transporting odor molecules entering the insect sensor lymph to the vicinity of the dendritic membrane of olfactory neurons, so that the odor receptors located on the membrane of neurons can recognize odor molecules and convert chemical signals into electrical signals to the central nervous system. Due to its principle, OBP and OBPP have high specificity for specific odor substances. However, OBP can only work normally under physiological conditions, so the application of OBP as a sensing material is limited to detecting substances in liquids. OBPP is a short peptide segment from the binding odor substance site in OBP, and the OBPP modified sensor can detect target odor substances in gas phase with low detection limit (LOD) and high selectivity. In recent years, through the use of OBP of cotton bollworm and fruit fly, OBPP sensors that can monitor octanal and isoamyl alcohol have been designed. These studies show the great potential of OBPP sensors. However, biosensors developed using odorant binding protein-derived peptides that bind pheromones (i.e., pheromone binding protein (PBP) derived peptides (Pheromone Binding Protein-derived Peptide, PBPP)) have not been reported. SUMMARY

[0004] Based on the above deficiencies, the purpose of the present application is to provide a pheromone binding protein-derived peptide of the American white moth, characterized in that the amino acid sequence of the pheromone binding protein-derived peptide is shown as SEQ ID NO: 2.

[0005] Another purpose of the present application is to provide a biosensor comprising an interdigital electrode, wherein single-walled carbon nanotubes are firmly attached to the interdigital electrode, characterized in that it further comprises a pheromone binding protein-derived peptide connected to the single-walled carbon nanotubes, and the amino acid sequence of the pheromone binding protein-derived peptide is shown as SEQ ID NO: 2.

[0006] Further, the interdigital electrode is 14 pairs of gold electrodes, the electrode width is 5 μm, the electrode spacing is 0.18 mm, the electrode thickness is 5 μm, the substrate material is Al2O3, and the device size is 10 mm x 20 mm x 1 mm.

[0007] The application also provides an electronic device for detecting the sex pheromone of Malacosoma neustria, comprising the biosensor as described above.

[0008] Advantages and beneficial effects of the application: the sensor of the pheromone binding protein derived peptide of the application can sensitively and specifically detect the concentration change of the sex pheromone of Malacosoma neustria, and has the ability to monitor female Malacosoma neustria in real time. BRIEF DESCRIPTION OF DRAWINGS

[0009] Figure 1 is the design schematic and flowchart of the pheromone binding protein derived peptide of Malacosoma neustria provided in the embodiment;

[0010] Figure 2 is the response graph of the pheromone binding protein derived peptide sensor to the sex pheromone of Malacosoma neustria provided in the embodiment;

[0011] a: no response graph of the sensor of unmodified PBPP to sex pheromone 1;

[0012] b: no response graph of the sensor of unmodified PBPP to sex pheromone 2;

[0013] c: no response graph of the sensor of unmodified PBPP to sex pheromone 3;

[0014] d: response graph of the PBPP2 sensor to sex pheromone 1 with a concentration of 500 ppb to 10 ppm;

[0015] e: no response graph of the PBPP2 sensor to sex pheromone 2;

[0016] f: response graph of the PBPP2 sensor to sex pheromone 3 with a concentration of 100 ppb to 10 ppm;

[0017] g: dose-dependent response curve graph of the PBPP2 sensor to sex pheromone 1 and sex pheromone 3;

[0018] Figure 3 is the stability, humidity and temperature influence and selectivity graph of the No. 2 pheromone binding protein derived peptide sensor to the No. 3 sex pheromone of Malacosoma neustria provided in the embodiment;

[0019] a: operation stability graph of the PBPP2 sensor responding to sex pheromone 3 with a concentration of 100 ppb;

[0020] b: storage stability graph of the PBPP2 sensor responding to sex pheromone 3 with a concentration of 1 ppm;

[0021] c: influence graph of the PBPP2 sensor responding to sex pheromone 3 with a concentration of 1 ppm at different temperatures;

[0022] d: PBPP2 sensor response graph to pheromone 3 at 1 ppm concentration at different humidity levels;

[0023] e: PBPP2 sensor selectivity graph to pheromone 3 (concentration of all substances is 500 ppb);

[0024] f: PBPP2 sensor selectivity graph to pheromone 3 (concentration of all substances is 100 ppb);

[0025] Figure 4 is the response graph of the pheromone binding protein derivative peptide sensor No. 2 provided by the present embodiment to the sex pheromone released by live American white moths:

[0026] a: No response graph of the unmodified PBPP sensor to the sex pheromone released by 50 female American white moths;

[0027] b: Response graph of the PBPP2 sensor to the sex pheromone released by 5, 10, 20, 50 female and 50 male American white moths. DETAILED DESCRIPTION

[0028] The present application will be further described below according to examples:

[0029] Example 1

[0030] Test materials and methods for preparing pheromone binding protein derivative peptide sensors

[0031] 1. Construction of the tertiary structure of the American white moth pheromone binding protein HcunPBP1

[0032] Figure 1 is the American white moth pheromone binding protein derivative peptide design schematic and flowchart provided by the present embodiment.

[0033] The coding region sequence of the pheromone binding protein HcunPBP1 that recognizes the sex pheromone of the American white moth was obtained according to the reported research. The tertiary structure of HcunPBP1 was obtained by searching and comparing the coding region sequence of the pheromone binding protein HcunPBP1 in the SWISS-MODEL website (https: / / swissmodel.expasy.org). The complete coding region amino acid sequence in FASTA format is as follows.

[0034] > HcunPBP1

[0035] MSVKLAILVAACLAIRVETSQDVIKQMAINFVKPLEACKKEMDIPETVIQDFYNFWKEGYELTNRQMGCAILCMSSKLELIDGEMNLHHGNAQEFAKKHGADDAMAKQLTDIIHNCVQTSPEAPDDPCQKTLNTVICFKAEIHKLNWAPNPELLVGELLAETK (as shown in SEQ ID NO: 4).

[0036] 2. Prediction of the binding sites of three sex pheromones recognized by the pheromone binding protein HcunPBP1 of Hyphantria cunea and design of PBPP sequences

[0037] Molecular docking was performed to predict the ligand binding sites using software Discovery studio 2016. First, the 3 kinds of molecules ((9z, 12z)-octadecadienal (sex pheromone 1, SP1), (9z, 12z, 15z)-octadecatrienal (sex pheromone 2, SP2), (3z, 6z)-cis-9, 10-epoxy-3, 6-nonadecadiene (sex pheromone 3, SP3), CAS numbers are 2541-61-9 (SP1), 2423-13-4 (SP2), 81309-90-2 (SP3), and the molecular conformation is shown in the supporting information. The cDOCKER function in DS2016 was used for precise docking, and the entire protein was defined as the receptor (HcunPBP1). Figure 1 The binding site was automatically defined from the receptor cavity, and site 1 was selected for one-step molecular docking, with a binding radius of 0.5. Repeat the operation to integrate all ligand binding sites. The docking results were visualized using software Pymol 2.6. According to the ligand binding site results, a polypeptide sequence of 7-11 aa in length containing adjacent ligand binding sites was selected as the sequence of PBPP. In order to be connected to the carbon nanotube, a cysteine (Cys, C) was added to the N-terminal.

[0038] The docking results gave a total of 10 conformations, and the docking parameters of the best four conformations, -CDOCKER_ENERGY, were 17.24 (HcunPBP1 vs. SP1), 1.16 (HcunPBP1 vs. SP2), and 14.79 (HcunPBP1 vs. SP3), respectively. The values of -CDOCKER_INTERACTION_ENERGY were 51.47 (HcunPBP1 vs. SP1), 43.19 (HcunPBP1 vs. SP2), and 51.15 (HcunPBP1 vs. SP3), respectively. The predicted binding sites are as follows:

[0039] HcunPBP1 recognition SP1 predicted 8 binding sites, Ala28, Phe31, Val32, Phe52, Phe55, Trp56, Met85, Lys130. There are 4 kinds of forces, respectively, traditional hydrogen bond, carbon hydrogen bond, Alkyl and π-alkyl. There are 2 traditional hydrogen bonds, the distance between atoms is Lys130 2.3 Å, Lys130 2.3 Å, 1 carbon hydrogen bond, the distance between atoms is Met85 2.5 Å.

[0040] HcunPBP1 recognition SP2 predicted 1 binding site, Leu87. The type of force is Alkyl, the distance between atoms is Leu87 5.0 Å.

[0041] HcunPBP1 recognition SP3 predicted 1 binding site, Leu87, Ala92, Leu109, Ile113, Val117. The force is Alkyl, the distance between atoms is Leu87 5.4 Å, Ala92 4.0 Å, Leu109 5.0 Å, Ile113 5.3 Å, Val117 3.8 Å. Based on these ligand binding site results, 3 PBPPs were designed, and the specific binding sites and sequences are CQDFYNFWKE (PBPP1, SEQ ID NO: 1), CEMNLHHGNAQ (PBPP2, SEQ ID NO: 2), CQLTDIIHNCVQ (PBPP3, SEQ ID NO: 3).

[0042] 3. Preparation of PBPP

[0043] Cysteine (Cys, C) residues were added to the N-terminus of the designed PBPP to achieve chemical connection of the peptide with single-walled carbon nanotubes (SWCNT) (Steglich esterification reaction (SER) and native chemical ligation (NCL)). The sequences of the synthesized PBPPs are CQDFYNFWKE (PBPP1, SEQ ID NO: 1), CEMNLHHGNAQ (PBPP2, SEQ ID NO: 2), CQLTDIIHNCVQ (PBPP3, SEQ ID NO: 3). PBPPs were synthesized by Hangzhou Special Peptide Biological Technology Co., Ltd. (Zhejiang, China). PBPPs were stored in a refrigerator at a stable temperature of -20 °C. PBPPs were dissolved to 0.01 mg / L with distilled water to connect SWCNTs.

[0044] 4. Preparation of gas samples

[0045] (9z, 12z)-octadecadienal (SP1), (9z, 12z, 15z)-octadecatrienal (SP2), (3z, 6z)-cis-9, 10-epoxy-3, 6-nonadecadiene (SP3), nonanal, hexanal, trans-2-hexen-1-al, 1-hexanol, isopentyl acetate, citronellal, cis-3-hexen-α-ol, trans-2-penten-1-ol, cis-2-penten-1-ol, dibutyl phthalate, linolenic acid, palmitic acid, phytol, d-limonene, camphene, a-phellandrene and pinene were purchased from KemaTech Chemicals Technology Co., Ltd. (Tianjin, China). Test gas standard samples were prepared from purchased test gas stock solutions (v / v, analytical grade). The required volume of test gas stock solution was calculated by the following equation:

[0046]

[0047] where V m is the molar volume of the gas (24500 mL / mol at 25 °C and one standard atmosphere), p is the density of the test gas (g / mL), V box (18000 mL) is the volume of the measuring chamber with the size of 30 (L) x 30 (W) x 20 (H) cm, M is the molar mass of the test gas (g / mol), C ss is the concentration of the purchased test gas stock solution (v / v). C ts is the target concentration of the test gas, V ts (mL) is the volume of the test gas stock solution required. V ts was calculated by equation (1). The standard solution of the required V ts was moved into a 100 mL sealed glass bottle using a microsyringe, and then evaporated. If the concentration was very low and the volume of the required standard substance could not be measured using a microsyringe, the evaporated standard substance could be diluted 10 times with air. In order to completely evaporate the standard substance in the container, the entire closed container was placed in an oven with a temperature of 10 °C higher than the boiling point of the target substance to be evaporated. After the solution was completely evaporated, the gas in the bottle was extracted and pumped into the test chamber using a 100 mL syringe.

[0048] 5. Preparation of PBPP sensor

[0049] First, the carboxylated SWCNTs were dispersed in pure ethanol and treated by ultrasonic oscillation for 20 minutes to prepare a suspension with a concentration of 0.1 mg / mL. Then, 0.5 mL of the carboxylated SWCNT suspension was taken and dropped onto the surface of an interdigital electrode. The interdigital electrode had an alumina (Al203) substrate and gold (Au) electrode material, and had the following structure parameters: 14 pairs of electrodes, an electrode spacing of 0.18 mm, a thickness of about 5 μm, a width of 5 μm, and an overall device size of 10 mm x 20 mm x 1 mm. After the ethanol was completely naturally volatilized, nitrogen was used for blowing treatment to remove the carboxylated SWCNTs that were not stably combined on the surface of the interdigital electrode. Next, a mixed reaction solution was prepared, which included 0.2 M N,N'-dicyclohexyl carbodiimide, 0.1 M benzyl mercaptan, and 0.05 M 4-dimethylaminopyridine, and the solvent was dichloromethane. The interdigital electrode loaded with the carboxylated SWCNTs was immersed in the above mixed solution for 2 hours of soaking treatment. Then, it was transferred to a solution containing 0.01 mg / L of a derivative peptide, and 0.05 M 4-mercaptophenol was used as a catalyst, and the reaction lasted for 48 hours. The sensor used resistance change as a detection signal, and when the concentration of the American white moth sex pheromone changed, the sensor could respond in real time and output the corresponding resistance change value.

[0050] 6. Gas detection

[0051] The sensor detected signals based on resistance change. It was placed in a sealed detection box, and a direct current probe (applied current 1 mA, applied voltage < 5 V) and a resistance analyzer (HPS2518, Changzhou Haierpa Electronics Technology Co., Ltd., Changzhou, China) were used to monitor the resistance value change in real time. After the baseline of the sensor was stable, a syringe was used to inject the target gas sample into the detection box at a flow rate of about 100 mL / s, and then the detection was sealed. The response value of the sensor to the pheromone compound was calculated by the formula ΔR / R0x 100%, where ΔR was the resistance change amount, and R0 was the initial resistance value. Different concentration gradients of gas samples were prepared, and the resistance response curve of the sensor was recorded in real time through multiple cycle dynamic tests. Each test cycle included an air injection stage and an air exhaust stage: the target gas was injected into the sealed detection chamber in the air injection stage, and fresh air was introduced for purification in the air exhaust stage. All test data were the average value ± standard deviation of three independent sensors, each measured three times. The whole experiment was completed in a constant temperature environment of 25°C, effectively controlling the influence of environmental temperature on the performance of the sensor.

[0052] Example 2

[0053] Detection of American white moth sex pheromone standard by pheromone binding protein derivative peptide sensor

[0054] Experimental grouping and experimental methods

[0055] Experiment group 1: The response of the sensor of unmodified PBPP (containing only SWCNT, gold electrode, alumina substrate) to sex pheromone 1. Experimental method: The real-time resistance value of the sensor was measured using a direct current resistance analyzer (HPS2518, Changzhou Haierpa Electronic Technology Co., Ltd.). After the resistance value of the sensor was stable, the initial resistance R0 was recorded, and then different concentrations of sex pheromone were pumped into the test box using a syringe; the resistance change rate AR / R0% (AR is the resistance change amount, and R0 is the initial resistance) was used as the response parameter of the sensor to sex pheromone 1.

[0056] Experiment group 2: The response of the sensor of unmodified PBPP to sex pheromone 2. Experimental method: The same as experiment group 1.

[0057] Experiment group 3: The response of the sensor of unmodified PBPP to sex pheromone 3. Experimental method: The same as experiment group 1.

[0058] Experiment group 4: The response of the sensor of modified PBPP2 to sex pheromone 1. Experimental method: The same as experiment group 1.

[0059] Experiment group 5: The response of the sensor of modified PBPP2 to sex pheromone 2. Experimental method: The same as experiment group 1.

[0060] Experiment group 6: The response of the sensor of modified PBPP2 to sex pheromone 3. Experimental method: The same as experiment group 1.

[0061] Experiment group 7: The response of the sensor of modified PBPP2 to sex pheromone 3 for five times. Experimental method: The response of the same PBPP2 sensor to 100 ppb of sex pheromone 3 was tested for five times in succession.

[0062] Experiment group 8: The response of the sensor of modified PBPP2 to sex pheromone 3 for seven days. Experimental method: The response of the same PBPP2 sensor to 1 ppm of sex pheromone 3 was tested at the same time every day for seven consecutive days.

[0063] Experiment group 9: The response of the sensor of modified PBPP2 to sex pheromone 3 at different temperatures. Experimental method: The response of the PBPP2 sensor to 1 ppm of sex pheromone 3 was tested at 23, 24, 25, 26, and 27°C, respectively.

[0064] Experiment group 10: The response of the sensor of modified PBPP2 to sex pheromone 3 at different humidities. Experimental method: The response of the PBPP2 sensor to 1 ppm of sex pheromone 3 was tested at 38, 39, 40, 41, and 42% RH, respectively.

[0065] Experiment group 11: The response of the sensor modified with PBPP2 to 20 substances at a concentration of 500 ppb. Experimental method: First, prepare the gas of sex pheromone 1, sex pheromone 2, sex pheromone 3, nonanal, hexanal, trans-2-hexen-1-al, 1-hexanol, isoamyl acetate, citronellal, cis-3-hexen-α-ol, trans-2-penten-1-ol, cis-2-penten-1-ol, dibutyl phthalate, linolenic acid, palmitic acid, phytol, d-limonene, camphene, a-phellandrene and pinene. Test the response of the PBPP2 sensor to these 20 substances in turn according to the experimental method of experiment group 1.

[0066] Experiment group 12: The response of the sensor modified with PBPP2 to 20 substances at a concentration of 100 ppb. Experimental method: Same as experiment group 11.

[0067] Experimental results

[0068] This example measures the change in resistance of the sensor in response to different concentrations of the sex pheromone sample of the American white moth. Seven different concentrations (5 ppb-10 ppm) of sex pheromones SP1, SP2 and SP3 are prepared, and the change in resistance of the sensor in response to the sex pheromone is monitored in real time through multiple cycles. The measurement period includes the gas-in state and the gas-out state. In the gas-in state, the sex pheromone gas is filled in a closed test box, and then purified with fresh air to achieve the gas-out state. When exposed to different concentrations of sex pheromones SP1, SP2 and SP3, the resistance of the sensor without PBPP modification remains unchanged, indicating that pure SWCNT, gold electrodes and alumina substrate have no response to sex pheromones SP1 ( Figure 2 , experiment group 1), SP2 ( Figure 2 b, experiment group 2) and SP3 ( Figure 2 c, experiment group 3). At the same time, the sensors modified with PBPP1 and PBPP3 do not respond to sex pheromones SP1, SP2 and SP3. On the other hand, the sensor modified with PBPP2 shows a response to sex pheromone SP1 ( Figure 2 d, experiment group), does not respond to sex pheromone SP2 ( Figure 2 e, experiment group 5), and shows a significant response to SP3 ( Figure 2 f, experiment group 6). This is likely due to the presence of two lone pair electrons on the oxygen atom of the aldehyde group of sex pheromone SP1, and the presence of two lone pair electrons on the oxygen atom of the epoxy group and the oxygen atom of the hydroxyl group of sex pheromone SP3. When the sex pheromone molecule binds to PBPP, the electrons fill the holes in the p-type semiconductor SWCNT, resulting in a decrease in carrier concentration and thus an increase in the resistance of the sensor.

[0069] Furthermore, the LOD values ​​of the PBPP2 sensor for SP1 and SP3 were compared. The LOD values ​​of the PBPP2 sensor for detecting SP1 and SP3 gases were 500 ppb and 100 ppb, respectively, with response ranges of 500 ppb to 5 ppm and 100 ppb to 5 ppm, respectively. Figure 2 (As shown in g). When the concentration of SP1 gas increased to 5 ppm and the concentration of SP3 gas increased to 5 ppm, the resistance change rate of the PBPP2 sensor reached a stable range, indicating that SP1 and SP3 gas molecules completely occupied the relevant binding sites of PBPP2 at the corresponding concentrations. Since SP3, the sex pheromone released by the fall webworm, accounts for approximately 99% of all sex pheromones, and the LOD (100 ppb) of PBPP2 for SP3 is lower than that for SP1 (500 ppb), it has a performance advantage, and therefore SP3 was chosen for focused testing.

[0070] Secondly, the operation of the PBPP2 sensor is stable ( Figure 3 a, Experimental group 7) and storage stability ( Figure 3 b, Experimental group 8) conducted experiments, and the sensor's response to the same sex pheromone concentration over seven days and five consecutive cyclic tests showed only slight amplitude variations, indicating that the sensor's performance is stable. Then, temperature ( Figure 3 c, Experimental group 9) and humidity ( Figure 3 The effects of experimental group 10 on the PBPP2 sensor were investigated. The results showed that changes in relative humidity and temperature had a certain monotonic effect on the sensor's resistance change rate.

[0071] Finally, the PBPP2 sensor's response to sex pheromones 1, 2, and 3, and 17 host volatiles was determined to establish selectivity (including nonanal, hexanal, trans-2-hexen-1-al, 1-hexanol, isoamyl acetate, citronellal, cis-3-hexen-α-ol, trans-2-penten-1-ol, cis-2-penten-1-ol, dibutyl phthalate, linolenic acid, palmitic acid, phytol, d-limonene, camphene, α-phellandrene, and pinene). The results showed that at 500 ppb, the PBPP2 sensor responded only to sex pheromones SP1 and SP3, and did not respond to any of the other 18 host volatiles. Figure 3 e, Experimental Group 11), at 100 ppb, the PBPP2 sensor only responded to the sex pheromone SP3 ( Figure 3f, Experimental Group 12). The PBPP2 sensor responded significantly more to sex pheromones SP1 and SP3 than to these 18 host volatiles, indicating that these two sensors have good specificity for sex pheromones. This is likely because PBPP is designed based on the biological interaction between receptors and pheromone molecules, and the binding occurs through a structure formed by amino acid residues surrounding the binding site. This binding is generally considered to be highly specific.

[0072] Example 3

[0073] PBPP sensors detect sex pheromones released by live female fall webworms.

[0074] Experimental Grouping and Experimental Methods

[0075] Experimental Group 13: Response of an unmodified PBPP sensor to sex pheromones released by live fall webworms. Experimental Method: First, the unmodified PBPP sensor was continuously tested in the dark for 0.5 hours. Then, 50 female moths were placed in a test chamber for 24 hours, with the lights turned on 16 hours after the moths were introduced to create a lit environment.

[0076] Experimental Group 14: Sensor response to sex pheromones released by live fall webworms. Experimental method: First, the PBPP2-modified sensor was continuously tested in the dark for 0.5 hours. Then, 5, 10, 20, and 50 female moths and 50 male moths were placed in the test chamber for 24 hours, respectively. The test environment was illuminated 16 hours after the moths were placed in the chamber.

[0077] Experimental results

[0078] To elucidate the potential field applications of the PBPP2 sensor, we tested its response to sex pheromones released by fall webworms one day after emergence. No response was observed in 50 female fall webworms to the unmodified PBPP2 sensor, indicating that the substrate did not respond to the moths. Figure 4 a, Experimental group 13). No obvious signals were observed from male fall webworms, indicating that male fall webworms did not release the sex pheromone SP3, and that other substances released by male fall webworms had no effect on the sensor. Figure 4 b, Experimental group 14). In contrast, the PBPP2 sensor showed a clear response to female fall webworms during the dark period, detecting as few as 5 females, and exhibited a greater change in electrical resistance as the number of adults one day after emergence increased. Figure 4 b, Experimental group 14). These results indicate that the PBPP2 sensor is suitable for real-time application and features high stability, sensitivity, and specificity, making it ideal for monitoring the early occurrence of pests.

[0079] For the same purpose, the present embodiment also provides an electronic device comprising the sensor described above.

Claims

1. A pheromone-binding protein-derived peptide that can effectively monitor the sex pheromone of the fall webworm, characterized in that, The amino acid sequence of the pheromone-binding protein-derived peptide is shown in SEQ ID NO:

2.

2. A biosensor capable of effectively monitoring the sex pheromones of the fall webworm, characterized in that, The device includes an interdigitated electrode on which a single-walled carbon nanotube is firmly attached, and the single-walled carbon nanotube is connected to a pheromone-binding protein-derived peptide, the amino acid sequence of which is shown in SEQ ID NO:

2.

3. The biosensor according to claim 2, characterized in that: The interdigitated electrodes consist of 14 pairs of gold electrodes with an electrode width of 5 μm, an electrode spacing of 0.18 mm, an electrode thickness of 5 μm, an Al2O3 substrate, and device dimensions of 10 mm × 20 mm × 1 mm.

4. An electronic device for detecting the sex pheromones of the fall webworm, characterized in that, Including the biosensor as described in claim 2 or 3.

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