Odor receptor gene OR49b recognizes benzothiazoles and its application in the control of Bactrocera dorsalis

Abstract

The present invention belongs to the technical field of pest control, and discloses the recognition of benzothiazole by the odor receptor gene OR49b and its application in the prevention and control of the fruit fly. Specifically, the application of the odor receptor BdorOR49b or the odor receptor gene BdorOR49b as a target in the prevention and control of the fruit fly is disclosed. The present invention discloses the key odor receptor OR49b of the fruit fly that senses the egg-laying attractant benzothiazole, and provides a reliable molecular target for the development of a new and efficient female or bisexual trap for the fruit fly, which can provide a new and feasible approach for the prevention and control of the fruit fly. At the same time, the basic data of the protein structure of OR49b and its relationship with the binding ability of benzothiazole can provide an important theoretical basis for the modification and effectiveness improvement of the fruit fly-specific trap targeting the key odor receptor.

Description

Technical Field

[0001] The invention belongs to the technical field of pest control, and particularly relates to an odor receptor gene OR49b for recognizing benzothiazole and an application thereof in the control of fruit fly dorsalis. Background Art

[0002] Bactrocera dorsalis belongs to the order Diptera, family Tetriphitidae. It has a wide range of food habits and causes serious damage to more than 150 kinds of fruits, including citrus, guava, mango, sweet cherry, Surinam cherry, loquat, papaya, etc. In recent years, due to its strong environmental adaptability, strong invasion ability, and rapid development of resistance, the damage caused has become increasingly serious. At present, the most effective prevention and control method is to use odor trapping combined with insecticides. For example, methyl eugenol is often used in the field to form an effective trapping technology for male citrus fruit flies. Since methyl eugenol itself does not have insecticidal ability and even has a certain promoting effect on the mating of male insects, its use not only requires the selection of suitable insecticide compounding, but also the consideration of factors that are not conducive to prevention and control, such as the enhancement of pest fertility. These factors will hinder the development of male insect trapping technology to a certain extent. At the same time, given that the damage of the orange fruit fly is mainly caused by the female adult laying eggs, there is no trap on the market that can effectively attract female or both sexes of the orange fruit fly. Therefore, researching and obtaining new traps to reduce the damage of the orange fruit fly has become an urgent problem to be solved. Summary of the invention

[0003] The purpose of the first aspect of the present invention is to provide the use of odorant receptor BdorOR49b or odorant receptor gene BdorOR49b as a target.

[0004] The second aspect of the present invention aims to provide a use of a BdorOR49b promoter in controlling the fruit fly and / or preparing a product for controlling the fruit fly.

[0005] The third aspect of the present invention aims to provide a method for regulating the sensitivity of Bactrocera dorsalis to benzothiazole.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] The first aspect of the present invention is to provide the use of odorant receptor BdorOR49b or odorant receptor gene BdorOR49b as a target in at least one of (1) to (6):

[0008] (1) Control of the fruit fly;

[0009] (2) Screening or preparing products for controlling Bactrocera dorsalis;

[0010] (3) Regulate the behavior of the fruit fly;

[0011] (4) preparing a behavior regulator for the fruit fly;

[0012] (5) preparing female fruit fly traps or female egg-laying attractants and bisexual traps;

[0013] (6) Regulate the sensitivity of Bactrocera dorsalis to irritating odor molecules.

[0014] In some embodiments of the present invention, BdorOR49b is used as a target to regulate the olfactory sensitivity of Bactrocera dorsalis to stimulating odor molecules.

[0015] In some embodiments of the present invention, the odor stimulating molecules include plant odors and their derivatives or insect sex pheromones and their analogs.

[0016] In some embodiments of the present invention, the odor stimulating molecule comprises benzothiazole.

[0017] The life cycle of the orange fruit fly is closely related to plant-derived compounds. They specifically identify volatiles from host plants and obtain or isolate these chemicals. Benzothiazole is a volatile detected in mangoes, which can significantly increase the egg-laying rate of female orange fruit flies and effectively lure both male and female insects. Since benzothiazole is considered to be one of the possible factors affecting the selection of egg-laying sites for orange fruit flies, it also indirectly reflects the potential attraction of this volatile to female orange fruit flies, but whether it has an attractant effect is still unclear. The present invention has been verified for the first time to find that benzothiazole has an attractant effect on both male and female insects, and it has been found that the odor receptor BdorOR49b mediates the recognition of benzothiazole by the orange fruit fly.

[0018]

[0019] The structural formula of the compound benzothiazole.

[0020] The second aspect of the present invention is to provide the use of the BdorOR49b promoter in controlling the fruit fly and / or preparing products for controlling the fruit fly.

[0021] In some embodiments of the present invention, the BdorOR49b promoter includes a substance that promotes the activity of BdorOR49b and / or a substance that increases the expression level of BdorOR49b.

[0022] In some embodiments of the present invention, the BdorOR49b promoter achieves the control effect on the oriental fruit fly by enhancing the olfactory sensitivity of the oriental fruit fly to irritating odor molecules.

[0023] In some embodiments of the present invention, the odor stimulating molecules include plant odors and their derivatives or insect sex pheromones and their analogs.

[0024] In some embodiments of the present invention, the odor stimulating molecule comprises benzothiazole.

[0025] The third aspect of the present invention is to provide a method for regulating the sensitivity of the oriental fruit fly to benzothiazole, comprising the step of expressing or knocking out the odor receptor gene BdorOR49b in the oriental fruit fly.

[0026] In some embodiments of the present invention, CRISPR / Cas9 technology is used to knock out the BdorOR49b gene of Bactrocera dorsalis.

[0027] The fourth aspect of the present invention provides a mutant of the odorant receptor BdorOR49b of the oriental fruit fly. Compared with the wild-type oriental fruit fly, the mutant has a 314 bp base deletion and a 5 bp base (GGCAT) insertion in the BdorOR49b gene, and terminates prematurely at amino acid position 118.

[0028] The beneficial effects of the present invention are:

[0029] The present invention reveals the key odor receptor OR49b of the fruit fly that senses the egg-laying attractant benzothiazole, providing a reliable molecular target for the development of new and efficient female or bisexual traps for the fruit fly, and can provide a new and feasible approach for the control of the fruit fly. At the same time, the basic data of the protein structure of OR49b and its relationship with benzothiazole binding ability can provide an important theoretical basis for the modification and effectiveness improvement of the fruit fly-specific trap targeting the key odor receptor. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The diagram shows the egg-laying and attracting effects of benzothiazole tested by egg-laying experiment and trap experiment; A is a schematic diagram of the single-choice egg-laying experiment, the single-plate double-choice egg-laying experiment and the olfactory trap, B is the number of eggs laid by wild-type female insects stimulated by benzothiazole in the single-choice experiment, C is the number of eggs laid by wild-type female insects stimulated by benzothiazole in the single-plate double-choice experiment, D is the wild-type female insect trapping experiment, and E is the wild-type male insect trapping experiment; in the figure, * represents P < 0.05, ** represents P < 0.01, *** represents P < 0.005, and **** represents P < 0.001.

[0031] Figure 2To verify in vitro that the odor receptor BdorOR49b mediates the recognition of benzothiazole by the fruit fly; wherein, A is a schematic diagram of the ectopic expression of BdorOR in the Drosophila ab3A neuron, B is a representative response spectrum of the UAS-BdorOR49b strain, C is a representative response trace of the OR22abGAL4 strain, D is the response of neurons expressing BdoroR49b to 60 compounds (n=4), with the strongest response to benzothiazole, E is a representative trace of the dose response of neurons expressing BdoroR49b, and F is the dose response of neurons expressing BdoroR49b to benzothiazole (n=13).

[0032] Figure 3 The figure shows the construction of mutant BdorOR49b; A is a schematic diagram of the gene structure of wild-type BdorOR49b knocked out by gene editing, and the target sequences of two sgRNAs (green) and PAM sequences (red) are shown on the exons. The obtained mutations include a 314bp nucleotide fragment deletion and a 5bp non-homologous nucleotide fragment insertion (blue) in the genome. Middle right: the genotypes of wild-type, heterozygous and homozygous individuals are distinguished by PCR and agarose gel electrophoresis. B is the protein sequence of the predicted BdorOR49b WT and mutant (314bp deletion and 5bp insertion) alleles.

[0033] Figure 4 BdorOR49b - / - and wild-type Bactrocera dorsalis electrophysiological responses to benzothiazole; wherein, A is a schematic diagram of EAG measurement, B is the EAG response of female wild-type and BdorOR49b mutant to different concentrations of benzothiazole, and C is the EAG response of male wild-type and BdorOR49b mutant to different concentrations of benzothiazole; in the figure, * represents P < 0.05, and ** represents P < 0.01.

[0034] Figure 5 benzothiazole mutant BdorOR49b - / - and wild-type Bactrocera dorsalis; wherein, A is a schematic diagram of the egg-laying selection and trap experiment of the wild-type and BdorOR49b mutant, B is the egg-laying experiment of the wild-type and BdorOR49b mutant, C is the female trapping experiment of the wild-type and BdorOR49b mutant, and D is the male trapping experiment of the wild-type and BdorOR49b mutant; in the figure, * represents P < 0.05.

[0035] Figure 6To verify in vitro that the odor receptors BdorOR7a-3 and BdorOR43a-2 are not involved in mediating the recognition of benzothiazole by the oriental fruit fly; wherein, A is the response of neurons expressing BdoroR7a-3 to 61 compounds (n=4), with the strongest response to ethyl crotonate and no response to benzothiazole; B is the response of neurons expressing BdoroR7a-3 to 64 compounds (n=4), with the strongest response to 2-ethyl-3,5,6-trimethylpyrazine and no response to benzothiazole. DETAILED DESCRIPTION

[0036] The present invention will now be described in detail with reference to specific embodiments, but the scope of the present invention is not limited thereto.

[0037] Unless otherwise specified, the materials and reagents used in this example were obtained from commercial sources.

[0038] Example

[0039] In order to better understand the essence of the present invention, the following describes the recognition of benzothiazole by the odor receptor gene OR49b and its application in the prevention and control of the citrus fruit fly in combination with experimental operations and experimental results.

[0040] 1. The egg-laying effect of benzothiazole on fruit flies was determined by an experimental method of measuring the egg-laying behavior of fruit flies to specific chemical substances. The specific experimental method is as follows:

[0041] Ten 16-day-old pregnant female flies were selected and placed in an 18 cm × 12.5 cm × 14 cm oviposition cage for the experiment. Agarose disks were used as ovulation-inducing devices to determine the egg-inducing effects of chemicals. Water (100 mL) and agarose (0.8 g) were mixed to form disks and heated to boiling in a microwave. The agarose was cooled at room temperature and waited for it to solidify. A series of doses (10 μg, 50 μg, 100 μg, 100 μg, 1000 μg) of benzothiazole were added. Hexane was used as a negative control. The oviposition device containing benzothiazole and hexane was placed in a cage containing females (n=10) for a selection test and allowed to lay eggs for 16 hours. Eggs were harvested from the oviposition device and the number of eggs for each treatment was counted. Five biological replicates were performed for each dose.

[0042] 2. The experimental method of measuring the tendency behavior of fruit flies to specific chemical substances was used to determine the attracting effect of benzothiazole on fruit flies. The specific experimental method is as follows:

[0043] Use an acrylic transparent insect cage with a size of 18cm×12.5cm×14cm, and place a 50mL conical bottle inside as an insect trap. The conical bottle stopper is made of silicone and has a circular through opening with a diameter of about 2cm. Use a 1000μL pipette tip as the adult to enter the pipeline. The tip of the pipette tip is cut off, and the entire pipeline is about 5cm long. Two conical bottle traps are placed in one to place the odor to be tested and the other to place the control solvent. The odor to be tested is dissolved in paraffin oil and can be diluted step by step to the concentration you need.

[0044] 3. Use single sensor recording experimental method to determine the electrophysiological response of transgenic fruit flies to benzothiazole.

[0045] Using the transgenic fruit fly empty neuron system, the fruit fly receptor is expressed, and then the nerve is recorded by electrophysiological means. If it can be activated by benzothiazole, it proves that the fruit fly receptor can sense benzothiazole in vitro. This embodiment uses the in vivo heterologous expression system fruit fly AB3 empty neuron to further verify the function of the odor receptor BdorOR49b. The transgenic fruit fly strain is constructed according to the standard procedure, and the specific operation is as follows: BdorOR49b is subcloned into the pUAST vector to construct an expression plasmid. The plasmid was injected into a specific fruit fly strain to express BdorOR49b on the third chromosome, and finally a UAS-BdorOR49b fruit fly homozygous strain was obtained. After hybridizing the 22ab GAL4 strain carrying the balancer with the UAS-BdorOR49b fruit fly strain, a homozygous expression strain (w+ / w+, UAS-BdorOR49b / UAS-BdorOR49b, 22ab GAL4 / 22abGAL4) was obtained. For specific methods, please refer to the literature De Fouchier, A., Walker III, WB, & Montagné, N. (2017). Functional evolution of Lepidoptera olfactory receptors revealed by deorphanization of a moth repertoire. Nat Commun 8: 15709, that is, a mutant strain in which BdorOR49b is expressed in Drosophila ab3, which can be stably inherited.

[0046] The primer sequences used to construct transgenic Drosophila are as follows:

[0047]

[0048] The homozygous mutant strains obtained by hybridization were used for electrophysiological experiments. The specific process is as follows:

[0049] (1) Fixation of transgenic fruit flies: Select transgenic fruit flies 3 to 5 days after eclosion for the experiment. First, use a blade to cut off the tip of the 20μL gun tip to an appropriate size, and slowly push a single adult into the gun tip, so that its head and antennae are exposed and its body is fixed in the gun tip to ensure that it cannot move freely. Use dental wax to fix the gun tip on a glass slide, and completely pull out and fix one of the antennae. Transfer to a Leica Z16APO high-resolution stereo microscope to observe the distribution of the sensory organs to be recorded, adjust the focus and antennae position, and place the target sensory organ in the center of the field of view for easy recording.

[0050] (2) Electrode preparation: Use a needle grinder to immerse the tip of the tungsten wire electrode in a 40% KNO2 solution and grind it repeatedly. Insert the ground reference electrode into the compound eye of the transgenic fruit fly, and insert the recording electrode connected to the preamplifier into the base of the target sensor until a stable electrical signal appears on the display, that is, the frequency of the neural signal is significantly higher than the noise signal. Signal recording: When recording electrophysiological signals, connect the end of the Pasteur tube to the silicone tube of the stimulator, insert the tip of the Pasteur tube into the odor mixing tube, and step on the foot pedal for stimulation. The odor delivery tube should be facing the antennae, about 2 cm apart. Each reaction signal is recorded for 10 seconds, and the odor stimulation starts 1 second later. At the same time, the time interval between two adjacent stimulations should be no less than 30 seconds, so that the antennal neurons can return to a spontaneous state.

[0051] (3) Data analysis: The neuronal response signal was amplified ten times by a preamplifier and transmitted to a computer via an analog-to-digital converter. The recorded signal graph was analyzed using Autospike 32 (Syntech, Germany) software. The number of neurons contained in the sensory organ was determined by the difference in the amplitude of the signal impulse (Spike). The magnitude of the response of the neurons in the sensory organ to the test odor was calculated by recording the number of signal impulses one second after the stimulation minus the number of signal impulses one second before the stimulation. The response value was expressed as mean ± standard error. The graph was drawn using GraphPad PRISM 6.0 software (San Diego, California, USA).

[0052] Further knocking out the receptor (BdorOR49b) in the fruit fly, if the behavioral and electrophysiological responses to benzothiazole are weakened, it is fully proved that this receptor is an important receptor for the fruit fly to sense benzothiazole and mediate its behavior. The gene editing experimental method was used to mutate the odor receptor BdorOR49b. The specific process is as follows: (1) Preparation of injection solution: The concentration of sgRNA injected into the orange fruit fly is 300ng / μL. Before injection, take 2.5μL of gRNA for each target and mix them, add 0.3μL of 5μg / μL Cas9 protein, and centrifuge at 12000rpm in a 4℃ centrifuge for 10min. (2) Embryo collection: Generally, orange fruit fly eggs are collected once every 10min, and placed on a homemade egg stand in a certain direction. A small amount of Halocarbon Oil is added to completely immerse the eggs and avoid movement of the eggs during injection. (3) Preparation of injection needle: Set the parameters according to the instructions of the needle puller to break the glass capillary. After breaking it, use a micro-sample needle to add 3 μL of injection solution to prevent the liquid from being sucked back when grinding the needle and causing the needle to be blocked; process the needle tip on the needle grinder to make it have an ideal tip. Then connect the injection needle to the microinjection instrument. (4) Microinjection of embryos: Place the stage with arranged embryos under the microscope, adjust the position of the injection needle under a precision optical microscope, and place the injection needle and the embryo on the same plane. Press the pedal to adjust the drop volume. Insert the needle tip into the tail of the embryo. Deliver the mixture into the embryo by pressing the pedal on the microinjection instrument. Inject 200 eggs each time. The nucleotide sequence of the wild-type coding gene of BdorOR49b is as follows:

[0053]

[0054] The primer sequences used for each target gRNA and verification mutant are as follows:

[0055]

[0056] 4. The electrophysiological response of fruit flies to benzothiazole was determined by recording antennal potential of fruit flies.

[0057] Use paraffin oil as solvent to prepare the test odor into a 100μg / μL stock solution, and store it in a -20℃ refrigerator. Before the test, the storage concentration is gradient diluted to prepare the odor solution of the required concentration, which can be prepared on the spot. Prepare filter paper strips, and drop 10μL of odor solutions (test group) and paraffin oil (control group) of different concentrations on the filter paper strips, push the filter paper strips completely into the Pasteur tube, seal the tube mouth with sealing film, and mark the corresponding odor concentration on the outside of the tube for use. Use surgical scissors to cut off the heads of the female and male adults of the orange fruit fly at the appropriate age (12 days) after emergence, prepare two glass electrodes filled with 2 / 3 volume of 0.1M KCl solution, carefully insert one glass electrode into the cut part of the head, and then fix it on the metal electrode, and fix the other glass electrode on the metal electrode. Under the microscope, one of the antennae of the fruit fly (a small section of the terminal antennae can be cut off or not cut off, just unify) is covered and connected, and the baseline displayed by the instrument connection software is observed. After the baseline is stable, add the test odor for measurement. During the test, the end of the Pasteur tube is connected to the silicone tube for air intake, and the tip is inserted into the ventilated metal hole. The foot pedal is stepped on for stimulation. The flow rate of the test odor stimulation is 10mL / s, the stimulation time is 0.2s, and the time interval between two adjacent stimulations is not less than 30s, so that the antennae can return to a stable state. The EAG signal was amplified using a 10× AC / DC headstage preamplifier (Syntech, The Netherlands) and then collected using an Intelligent Data Acquisition Controller (IDAC-4-USB, Syntech, The Netherlands). Each antenna was stimulated twice with paraffin oil (the first and last time), and the order of the tested odor stimuli was from low to high concentration.

[0058] result:

[0059] Behavioral responses of Bactrocera dorsalis to benzothiazoles. We tested the oviposition-inducing effects of benzothiazoles on females in an egg-laying assay and the attractiveness of benzothiazoles to males and females in an olfactory trap assay ( Figure 1 In the double-choice oviposition and single-choice oviposition experiments, we found that female B. dorsalis laid more eggs in the area coated with benzothiazole than in the control area, and there was a significant difference in the number of eggs laid in different concentration treatments ( Figure 1In the two-choice experiment, they preferred the area containing benzothiazole and laid more eggs ( Figure 1 From 18:00 to 09:00, the olfactory trap method was used to observe the tropism of adult insects to benzothiazole at concentrations of 10, 50, 100, 500, 10000, and 10000 μg ( Figure 1 Benzothiazole in the range of 500 to 10,000 μg can effectively trap female adults ( Figure 1 D), 50-10000 μg of benzothiazole can effectively trap male adults ( Figure 1 In addition, the tropism of adults to the compound showed a dose-response ( Figure 1 D and E).

[0060] The transgenic fruit fly system was used to express the oriental fruit fly odorant receptor BdorOR49b for in vitro functional studies. Odorant receptors sensitive to similar structural fused heterocyclic compounds (indole) belong to a unique branch of mosquitoes and Drosophila melanogaster. Using phylogenetic tools, we found that oriental fruit fly BdorOR43a and BdorOR49b also belong to this branch. Fused heterocyclic organic compounds can serve as inter-kingdom signals to regulate communication between species. Benzthiazole is a derivative of fused heterocyclic compounds, so we cloned the full-length sequences of the receptors BdorOR43a (AKI29035.1) and BdorOR49b (XP_049309039.1) and further performed functional identification. We used transgenic technology to heterologously express the alleles BdorOR43a and BdorOR49b into Drosophila melanogaster ( Figure 2 We then examined the responses of ab3 neurons expressing BdorOR43a and BdorOR49b to benzothiazole. Transgenic flies expressing BdorOR43a and BdorOR49b produced correct spontaneous potentials, indicating that the ectopically expressed BdorORs were correctly localized. Among them, neurons expressing BdorOR49b showed a strong response to benzothiazole ( Figure 2 C to D), while neurons expressing BdorOR43a did not respond to benzothiazole. We defined the response spectrum of BdorOR49b using 60 chemicals, including the most common plant odors and derivatives (such as 1-octen-3-ol, 3-carene, methyl anthranilate, etc.), as well as insect sex pheromones and analogs (such as 2,3,5-trimethylpyrazine, 2-ethyl-3,5,6-trimethylpyrazine, etc.). BdorOR49b responded most strongly to benzothiazole ( Figure 2 Next, a dose-response study was performed in which BdorOR49b was activated by benzothiazole concentrations ranging from 1 to 1000 μg ( Figure 2(E) The EC50 value of BdorOR49b for benzothiazole is 4.249*10 -5 g(n>10 Figure 2 F). We further investigated whether the odorant receptors BdorOR7a-3 and BdorOR43a-2 mediate the recognition of benzothiazole by Bactrocera dorsalis. The results showed that neurons expressing BdoroR7a-3 responded to 61 compounds (n=4), with the strongest response to ethyl crotonate and no response to benzothiazole ( Figure 6 Middle A); The responses of neurons expressing BdoroR7a-3 to 64 compounds (n=4) were strongest to 2-ethyl-3,5,6-trimethylpyrazine, but not to benzothiazole ( Figure 6 (middle B).

[0061] In vivo function study of the odorant receptor BdorOR49b of the oriental fruit fly. We used CRISPR / Cas9 technology to knock out the BdorOR49b gene. sgRNAs (300 ng / μL, see the primer sequence information table for each target gRNA and verified mutant) and Cas9 protein (300 ng / μL, TrueCut TM The complex composed of Cas9 Protein v2, Catalog number: A36498) was injected into 200 eggs of the orange fruit fly, 84 eggs successfully hatched, and 61 larvae emerged as adults. Then the genome was extracted from a mid-leg of the injected G0 adult for genotyping. In terms of genetic hybridization, G0 was reversely hybridized with the wild type. Among G1, the G1 mutant carrying the same deletion was selected for subsequent experimental studies. After three generations of screening, a mutant with a long fragment deletion ( Figure 3 A). The 314 bp deletion and 5 bp insertion (sequence: GGCAT) caused a frameshift mutation in the BdorOR49b gene, leading to premature termination at amino acid position 118 and producing a non-functional BdorOR49b protein ( Figure 3 In B). To verify the function of BdorOR49b in vivo, we wanted to confirm whether the mutant had lost the ability to sense volatile chemicals by EAG (Electroantennography) detection. The EAG experiment showed that wild-type females and males had obvious EAG responses to benzothiazole, and the response trend showed a dose response. Four concentrations of benzothiazole (0.1-100 μg / L) could induce the electrophysiological response of BdorOR49b. After knocking out the BdorOR49b gene, the mutant's EAG response to benzothiazole was significantly reduced ( Figure 4In addition, we used egg-laying tests and olfactory trapping to detect the behavioral differences between BdorOR49b knockout individuals and wild-type individuals towards benzothiazole. The egg-laying test results showed that wild-type females successfully laid eggs, but BdorOR49b knockout females lost their egg-laying preference for benzothiazole ( Figure 5 (B). All concentrations of benzothiazole significantly reduced the attractiveness of BdorOR49b mutant males and females compared to wild type ( Figure 5 (C~D).

[0062] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. Application of odorant receptor BdorOR49b or odorant receptor gene BdorOR49b as a target in regulating the olfactory sensitivity of Bactrocera dorsalis to irritating odor molecules, wherein the irritating odor molecule is benzothiazole; the nucleotide sequence of BdorOR49b is shown in SEQ ID NO:

11.

2. Application of overexpression of BdorOR49b in controlling Bactrocera dorsalis; The overexpression of BdorOR49b achieves the control effect of the fruit fly by enhancing the olfactory sensitivity of the fruit fly to irritating odor molecules; The irritating odor molecule is benzothiazole; The nucleotide sequence of BdorOR49b is shown in SEQ ID NO:

11.

3. A method for regulating the sensitivity of Bactrocera dorsalis to benzothiazole, comprising the step of expressing or knocking out the odor receptor gene BdorOR49b in Bactrocera dorsalis; The nucleotide sequence of BdorOR49b is shown in SEQ ID NO:11; CRISPR / Cas9 technology was used to knock out the odorant receptor gene BdorOR49b of the fruit fly; The nucleotide sequence of the sgRNA involved in the CRISPR / Cas9 is shown in SEQ ID: 5 to SEQ ID NO: 8.

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