Use of olfactory receptors for identifying predetermined compounds in sample

Abstract

The invention relates to the technical field of biosensors, and particularly discloses application of an olfactory receptor in identification of a predetermined compound in a sample. The olfactory receptor comprises at least one selected from the group consisting of OR14I1, OR2T6, OR5H15, OR2F2 and the like, and the predetermined compound comprises at least one selected from the group consisting of isobornyl formate, L-borneol, 1-butyl-2-isobutyl phthalate, 2, 3-dihydro-2-methylbenzofuran, citral, diisobutyl phthalate, eugenol acetate, citronellyl formate and the like. The invention also provides recombinant cells for expressing the olfactory receptors and the reporter proteins, a kit containing the recombinant cells, and a method for identifying plant varieties by using the olfactory receptors, the recombinant cells or the kit. According to the method, the plant variety can be quickly and accurately identified by detecting the signal generated by activating the specific olfactory receptor by the volatile components of the plant, and the method has the advantages of high specificity and sensitivity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biosensors, and in particular, to the use of olfactory receptors in identifying a predetermined compound in a sample, more particularly to a recombinant cell, a kit for identifying a predetermined compound in a sample, a set of olfactory receptors for identifying a plant variety, and a method for identifying a plant variety. BACKGROUND

[0002] Mammals have excellent olfactory perception ability, and can sensitively and quickly identify and distinguish odors in food, individuals or environment. According to surveys, the prevalence of self-reported olfactory dysfunction is 3% to 15% in recent years, and it gradually increases with age. As a common but underestimated health problem, olfactory dysfunction is affecting the quality of life of hundreds of millions of people worldwide, and even endangering life safety.

[0003] Current treatment methods for olfactory dysfunction are diverse. Olfactory training, as a new treatment method in recent years, can increase olfactory sensitivity and improve the quality of life of patients, but its treatment mechanism is still unclear, and it is also unclear what the best combination of smells for olfactory training is. At the same time, the types of olfactory training agents are diverse, including medicinal materials, spices, essential oils, etc., which are often natural plant products with special smells. However, the authenticity of natural plant products on the market is difficult to guarantee, and the quality is uneven, which not only affects the rights and interests of consumers, but also may cause safety hazards. Therefore, more reliable and efficient identification technology is urgently needed to ensure that the source of the olfactory agent is "real, high-quality, stable, and controllable".

[0004] Olfactory receptors (ORs) play an important connecting role between odor molecules and olfaction. Combining high-throughput volatile component analysis with olfactory receptor target screening can not only promote the understanding of olfactory perception mechanisms, provide personalized treatment options for patients with olfactory dysfunction, but also help perfume, cosmetic and food companies develop products that better meet consumer preferences, and provide more reliable and difficult-to-fake identification methods for high-value natural products. SUMMARY

[0005] The present application aims to at least partially solve at least one of the technical problems existing in the prior art. To this end, the present application provides the use of olfactory receptors in identifying a predetermined compound in a sample.

[0006] The present application is based on the following findings of the inventors: The inventors, for the first time, systematically identified and verified 34 "compound-olfactory receptor" interaction pairs from 6 natural plant samples by combining SPME-GC-MS volatile component analysis technology with a high-throughput olfactory receptor functional screening platform. More importantly, among the above pairs, the inventors first discovered and verified 23 new "compound-olfactory receptor" interaction pairs, which have not been reported in the prior art. As a breakthrough contribution of the present application, the inventors successfully achieved the first "de-orphanization" of 4 olfactory receptors, i.e., the first identification of functional ligand compounds for orphan receptors OR14I1, OR2T6, OR5H15 and OR2F2. Based on the above findings, the present application not only expands the human olfactory receptor ligand atlas and provides new target information for understanding the olfactory perception mechanism, but also thereby constructs a unique fingerprint based on "volatile component-receptor activation signal", laying a solid foundation for realizing accurate and reliable identification of natural plant samples and developing targeted olfactory agents for olfactory training or treatment of olfactory disorders.

[0007] In one aspect of the present application, the present application provides use of an olfactory receptor in identifying a predetermined compound in a sample. According to embodiments of the present application, the olfactory receptor comprises at least one selected from the group consisting of OR14I1, OR2T6, OR5H15, and OR2F2; and the predetermined compound comprises at least one selected from the group consisting of isobornyl formate, levoborneol, 1-butyl 2-isobutyl o-phthalate, 2,3-dihydro-2-methylbenzofuran, citral, diisobutyl phthalate, eugenyl acetate, and citronellyl formate. The use according to embodiments of the present application has the remarkable effect of first time successfully applying a newly discovered set of specific "compound-olfactory receptor" pairings to construct a novel natural plant identification method. Specifically, the core contribution of the present application lies in completing the "de-orphaning" and functional definition of key olfactory receptors such as OR14I1, OR2T6, OR5H15, and OR2F2, and systematically constructing their corresponding identification relationships with a series of plant-derived volatile compounds such as isobornyl formate and eugenyl acetate. Based on this, we can convert complex plant chemical composition information into specific olfactory receptor-mediated biological activation signals that can be qualitatively and quantitatively determined. This conversion brings fundamental technical advantages: natural plant samples of different sources will activate a set of characteristic olfactory receptors due to differences in their volatile component compositions, thereby forming their unique "biological activation fingerprints". By analyzing whether the sample to be tested can activate core receptor combinations such as OR2T6 / OR5H15 (indicating eugenyl acetate) or OR14I1 (indicating diisobutyl phthalate), and comparing their activation profiles with standard profiles, the identification of plant species, authenticity, or origin can be achieved. This functional biological recognition-based strategy has higher inherent specificity and anti-matrix interference ability compared to simple chemical component analysis, and provides a more reliable new approach for accurate identification of natural plants.

[0008] In some aspects of the present application, the predetermined compound comprises at least one selected from the group consisting of diisobutyl phthalate and eugenyl acetate.

[0009] In some aspects of the present application, the olfactory receptor further comprises at least one selected from the group consisting of OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR12D2, OR5T2, OR4D2, OR51A7, and OR2J3.

[0010] In some aspects of the application, the OR2W1 is used to recognize at least one of the iso-bornyl formate and the laevo-borneol; or the OR2C1 is used to recognize the laevo-borneol; or the OR2M3 is used to recognize the laevo-borneol; or the OR2T11 is used to recognize the laevo-borneol; or the OR10X1 is used to recognize the laevo-borneol; or the OR52L1 is used to recognize the laevo-borneol; or the OR7A17 is used to recognize the laevo-borneol; or the OR9I1 is used to recognize the phthalic acid-1-butyl ester-2-isobutyl ester; or the OR5K1 is used to recognize the 2,3-dihydro-2-methylbenzofuran; or the OR3A3 is used to recognize at least one of the citral and the diisobutyl phthalate; or the OR7E24 is used to recognize the citral; or the OR14A2 is used to recognize the diisobutyl phthalate; or the OR14I1 is used to recognize the diisobutyl phthalate; or the OR12D2 is used to recognize the diisobutyl phthalate; or the OR5T2 is used to recognize the diisobutyl phthalate; or the OR4D2 is used to recognize the diisobutyl phthalate; or the OR2T6 is used to recognize the eugenyl acetate; or the OR5H15 is used to recognize the eugenyl acetate; or the OR2F2 is used to recognize the eugenyl acetate; or the OR51A7 is used to recognize the eugenyl acetate; or the OR2J3 is used to recognize the citronellyl formate.

[0011] In some aspects of the application, the olfactory receptor has an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 22.

[0012] In some aspects of the application, the OR14I1 has an amino acid sequence as set forth in SEQ ID NO: 14.

[0013] In some aspects of the application, the OR2T6 has an amino acid sequence as set forth in SEQ ID NO: 18.

[0014] In some aspects of the application, the OR5H15 has an amino acid sequence as set forth in SEQ ID NO: 19.

[0015] In some aspects of the application, the OR2F2 has an amino acid sequence as set forth in SEQ ID NO: 21.

[0016] In some aspects of the application, the OR2W1 has an amino acid sequence as set forth in SEQ ID NO: 1.

[0017] In some aspects of the application, the OR2C1 has an amino acid sequence as set forth in SEQ ID NO: 2.

[0018] In some aspects of the application, the OR2M3 has an amino acid sequence as set forth in SEQ ID NO: 3.

[0019] In some aspects of the application, the OR2T11 has an amino acid sequence as set forth in SEQ ID NO: 4.

[0020] In some aspects of the application, the OR10X1 has an amino acid sequence as set forth in SEQ ID NO: 5.

[0021] In some aspects of the application, the OR52L1 has an amino acid sequence as set forth in SEQ ID NO: 6.

[0022] In some aspects of the application, the OR7A17 has an amino acid sequence as set forth in SEQ ID NO: 7.

[0023] In some aspects of the application, the OR9I1 has an amino acid sequence as set forth in SEQ ID NO: 8.

[0024] In some aspects of the application, the OR5K1 has an amino acid sequence as set forth in SEQ ID NO: 9.

[0025] In some aspects of the application, the OR3A3 has an amino acid sequence as set forth in SEQ ID NO: 11.

[0026] In some aspects of the application, the OR7E24 has an amino acid sequence as set forth in SEQ ID NO: 12.

[0027] In some aspects of the application, the OR14A2 has an amino acid sequence as set forth in SEQ ID NO: 13.

[0028] In some aspects of the application, the OR12D2 has an amino acid sequence as set forth in SEQ ID NO: 15.

[0029] In some aspects of the present application, the OR5T2 has an amino acid sequence as set forth in SEQ ID NO: 16.

[0030] In some aspects of the present application, the OR4D2 has an amino acid sequence as set forth in SEQ ID NO: 17.

[0031] In some aspects of the present application, the OR51A7 has an amino acid sequence as set forth in SEQ ID NO: 22.

[0032] In some aspects of the present application, the OR2J3 has an amino acid sequence as set forth in SEQ ID NO: 20.

[0033] In some aspects of the present application, the sample is a plant sample.

[0034] In a second aspect of the present application, the present application provides a recombinant cell. According to an embodiment of the present application, the recombinant cell expresses an olfactory receptor and a reporter protein; the olfactory receptor comprises at least one selected from the group consisting of OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7 and OR2J3; the reporter protein is suitable for generating a detectable signal after the olfactory receptor recognizes the predetermined compound. According to the recombinant cell of the embodiment of the present application, by combining the newly discovered olfactory receptor of the present application with the reporter protein system, a high-efficiency, standardized biosensor core element is successfully constructed; this design directly converts the specific recognition behavior of specific odor molecules and receptors into a universal, quantifiable optical or electrochemical signal, thereby simplifying the complex chemical detection problem into an intuitive signal reading process, not only providing a stable, reusable detection tool for precise identification of natural plants, but also laying a solid foundation for realizing high-throughput, automated odor fingerprint analysis.

[0035] In some aspects of the present application, the olfactory receptor is expressed on the cell membrane of the recombinant cell.

[0036] In a third aspect, the present application provides a kit for identifying a predetermined compound in a sample. According to embodiments of the present application, the kit comprises the recombinant cell of the second aspect. According to embodiments of the present application, the kit can standardize the detection capability of the recombinant cell and productize the integration, and provide a complete solution ready for use. The kit can combine the recombinant cells expressing different olfactory receptors in a reasonable manner or be packaged separately, and necessary detection reagents are provided, so that even non-professionals can quickly and accurately complete the synchronous detection and identification of multiple characteristic odor molecules in complex samples under routine experimental conditions, greatly improving the operability and repeatability of the detection method based on olfactory receptors, and providing key support for the practical popularization and application of natural plant identification technology.

[0037] In a fourth aspect, the present application provides a set of olfactory receptors for identifying plant varieties. According to embodiments of the present application, the olfactory receptors comprise at least one selected from OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7 and OR2J3. According to embodiments of the present application, the core effect of the olfactory receptors is to provide a set of functionally verified and special receptor combinations for constructing a plant variety identification system. Each member of the receptor set has been verified to be able to specifically respond to one or more plant characteristic volatile compounds, such as the key pair of OR2T6 and eugenyl acetate, OR14I1 and diisobutyl phthalate. When these receptors are used as a whole, they are no longer isolated detection targets, but together constitute the core elements of a "biological sensor array" that can decode complex plant odor fingerprints. By selectively combining different members of the receptor set, the multi-dimensional biological response signal of the plant to be tested can be systematically collected, thereby establishing a unique identification model that can accurately distinguish different plant varieties. The establishment of this set of receptors provides the most fundamental and optimized material basis for the development of various olfactory receptor-based identification products (including but not limited to recombinant cells, kits and detection methods), ensuring the high specificity and reliability of the final identification results.

[0038] In a fifth aspect, the present application provides a method for identifying plant varieties. According to embodiments of the present application, the method comprises detecting a plant sample using the recombinant cell of the second aspect, the kit of the third aspect, or the olfactory receptors of the fourth aspect; and determining the variety of the plant sample based on the detection result.

[0039] In some aspects of the present application, the method comprises: stimulating the olfactory receptor in the recombinant cell of the second aspect of the present application or the kit of the third aspect of the present application or the olfactory receptor of the fourth aspect of the present application with the volatile component of the plant variety to be tested, obtaining a response value; determining whether the volatile component of the plant variety to be tested can activate the olfactory receptor based on the response value; determining the volatile component of the plant variety to be tested based on the category of the activated receptor; and determining the variety of the plant to be tested based on the category of the volatile component of the plant variety to be tested.

[0040] In some aspects of the present application, the response value being not lower than a preset threshold value indicates that the volatile component of the plant variety to be tested can activate the olfactory receptor.

[0041] In some aspects of the present application, the OR2W1 having an activation signal indicates that the volatile component of the plant variety to be tested has at least one of formyl isobornyl ester and levoborneol.

[0042] In some aspects of the present application, the OR2C1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0043] In some aspects of the present application, the OR2M3 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0044] In some aspects of the present application, the OR2T11 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0045] In some aspects of the present application, the OR10X1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0046] In some aspects of the present application, the OR52L1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0047] In some aspects of the present application, the OR7A17 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0048] In some aspects of the present application, the OR9I1 having an activation signal indicates that the volatile component of the plant variety to be tested has 1-butyl-2-isobutyl phthalate.

[0049] In some aspects of the present application, the OR5K1 having an activation signal indicates that the volatile component of the plant variety to be tested has 2,3-dihydro-2-methylbenzofuran.

[0050] In some aspects of the application, OR3A3 has an activation signal that is an indication that the volatile components of the plant variety under test have at least one of citral and diisobutyl phthalate.

[0051] In some aspects of the application, OR7E24 has an activation signal that is an indication that the volatile components of the plant variety under test have citral.

[0052] In some aspects of the application, OR14A2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0053] In some aspects of the application, OR14I1 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0054] In some aspects of the application, OR12D2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0055] In some aspects of the application, OR5T2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0056] In some aspects of the application, OR4D2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0057] In some aspects of the application, OR2T6 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0058] In some aspects of the application, OR5H15 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0059] In some aspects of the application, OR2F2 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0060] In some aspects of the application, OR51A7 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0061] In some aspects of the application, OR2J3 has an activation signal that is an indication that the volatile components of the plant variety under test have citronellyl formate.

[0062] In some aspects of the application, the volatile components of the plant variety under test have at least one of citronellyl formate and levorotatory borneol is an indication that the plant under test is a genus of genus Cinnamomum.

[0063] In some aspects of the application, the volatile components of the plant variety under test having 2,3-dihydro-2-methylbenzofuran are indicative of the plant under test being a Syzygium species.

[0064] In some aspects of the application, the volatile components of the plant variety under test having 2,3-dihydro-2-methylbenzofuran are indicative of the plant under test being a Syzygium species.

[0065] In some aspects of the application, the volatile components of the plant variety under test having 2,3-dihydro-2-methylbenzofuran are indicative of the plant under test being a Syzygium species.

[0066] In some aspects of the application, the volatile components of the plant variety under test having citral are indicative of the plant under test being a Zingiber species.

[0067] In some aspects of the application, the volatile components of the plant variety under test having diisobutyl phthalate are indicative of the plant under test being an Agastya species.

[0068] In some aspects of the application, the volatile components of the plant variety under test having eugenyl acetate are indicative of the plant under test being a Myristica species.

[0069] In some aspects of the application, the volatile components of the plant variety under test having citral are indicative of the plant under test being a Zingiber species.

[0070] Additional aspects and advantages of the application will be set forth in part in the description which follows, and in part will become apparent to those skilled in the art upon examination of the following and the accompanying drawings or can be learned by practice of the application. BRIEF DESCRIPTION OF DRAWINGS

[0071] The foregoing and / or additional aspects and advantages of the application will become apparent to those skilled in the art from the following description, which proceeds with reference to the accompanying drawings. Figure 1 Representative total ion chromatogram of SPME-GC-MS of natural plant sample in Example 2.

[0072] Figure 2 Representative total ion chromatogram of SPME-GC-MS of natural plant sample in Example 2.

[0073] Figure 3 Dose-dependent curve of functional compound interaction with olfactory receptors in Sample 1 in Example 3.

[0074] Figure 4 Dose-dependent curve of functional compound interaction with olfactory receptors in Sample 2 in Example 3.

[0075] Figure 5To obtain a dose-dependent curve of the functional compound of sample 3 in Example 3 interacting with olfactory receptors.

[0076] Figure 6 To obtain a dose-dependent curve of the functional compound of sample 4 in Example 3 interacting with olfactory receptors.

[0077] Figure 7 To obtain a dose-dependent curve of the functional compound of sample 5 in Example 3 interacting with olfactory receptors.

[0078] Figure 8 To obtain a dose-dependent curve of the functional compound of sample 5 in Example 3 interacting with olfactory receptors.

[0079] Figure 9 To obtain a dose-dependent curve of the functional compound of sample 6 in Example 3 interacting with olfactory receptors. DETAILED DESCRIPTION

[0080] Embodiments of the present application are described in detail below. The embodiments described below are examples only and are not intended to limit the present application, as interpreted in its broadest form.

[0081] It should be noted that the terms "first", "second", etc. are used only for descriptive purposes and are not to be construed as indicating or implying relative importance or a quantity of the specified technical features indicated. Thus, a feature defined with "first", "second", etc. can include one or more of the features implicitly or explicitly. Further, in the description of the present application, the meaning of "a plurality of" is two or more unless otherwise specified.

[0082] In order to facilitate the understanding of the present application, certain technical and scientific terms are specifically defined below. Unless otherwise apparent in the context of the present application, all other technical and scientific terms used in the present application have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs.

[0083] In the present application, the term "comprising" or "including" is an open expression, i.e., including the content indicated by the present application, but not excluding other aspects.

[0084] In the present application, the term "optionally", "optional" or "optional" generally means that the event or condition described subsequently can but need not occur, and the description includes cases where the event or condition occurs, as well as cases where it does not occur.

[0085] In the present invention, the term "olfactory receptor" refers to a class of G protein-coupled receptors (GPCRs) present on the cell membrane of olfactory sensory neurons, responsible for recognizing and binding odor molecules in the environment, and is the first step in olfactory perception. In this application, it specifically refers to human olfactory receptors, whose representative amino acid sequences are shown in SEQ ID NO: 1 to SEQ ID NO: 27. That is, the target and detection object of the present invention.

[0086] In the present invention, the term "SPME-GC-MS" refers to an analytical technique that integrates sample extraction, enrichment, separation, and identification. Among them, "SPME" refers to the use of solid-phase microextraction to non-destructively collect volatile components in the headspace of the sample, "GC" refers to the high-efficiency separation of complex mixtures, and "MS" refers to the qualitative identification and quantitative analysis of separated compounds. That is, the core analysis method used in the present invention to obtain the "volatile component spectrum" of natural plants.

[0087] In the present invention, the term "predetermined compound" refers to a group of volatile organic compounds pre-selected in the present invention, which can be recognized and activated by specific olfactory receptors (Olfactory Receptors, ORs), thereby used for detecting, identifying or analyzing specific chemical components in samples (such as plant samples). The core of this term is to emphasize that these compounds are pre-selected target molecules known to form specific pairs with olfactory receptors for constructing detection methods based on biological responses.

[0088] In the present invention, the term "key compound" refers to a compound selected from the volatile component spectrum of a natural plant sample according to pre-set standards for subsequent olfactory receptor screening. The volatile component spectrum is obtained by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS), and the relative percentage content of each compound is calculated by peak area normalization. The selection criteria are that the relative percentage content is at least greater than 0.4%, preferably greater than 0.5%, and is easily commercially available. This threshold ensures the significant presence of the compound in the sample, while improving the efficiency of the screening and the feasibility of the experiment.

[0089] Those skilled in the art should understand that in the present invention, the terms "predetermined compound" and "key compound" are synonymous terms, both referring to a set of compounds selected from natural plant samples by the above-mentioned SPME-GC-MS technique and standards, which can be specifically recognized and activated by specific olfactory receptors. The term "key compound" focuses on its screening source and initial definition, while the term "predetermined compound" focuses on its final role and function in the technical solution. Both terms in this application refer to the same material entity, and the equivalence is intended to ensure consistency in the terminology of the specification and claims, thereby clearly defining the scope of protection.

[0090] In the present invention, the term "functional compound" refers to those compounds that have been proven to be able to specifically activate one or more particular olfactory receptors in olfactory receptor screening, and the activation effect thereof presents significant dose dependence. They are the active part of "key compounds" with biological activity. That is, the active molecules with clear biological function discovered in the present invention are the core of constructing "ligand-receptor" pairs.

[0091] In the present invention, the term "biomimetic olfactory receptor cell library" refers to a cell collection containing multiple (such as 400) human olfactory receptors and downstream signal pathway elements (Golf protein, reporter gene, etc.) that are heterologously expressed in an in vitro cell model (such as HEK293T cells) by molecular biology methods (such as co-transfection). The cell library can simulate the response of in vivo olfactory neurons to odor molecules. That is, the present invention realizes the core technology platform and tool for high-throughput functional screening of olfactory receptors.

[0092] In the present invention, the term "Dual-Glo TM Dual-luciferase system" refers to a high-sensitivity gene reporter system for detecting the degree of activation of a specific signal pathway (such as the cAMP pathway) in cells. The system simultaneously uses firefly luciferase (as the main reporter gene, responding to pathway activation) and sea pansy luciferase (as the internal reference gene, for normalization, to eliminate the interference of factors such as transfection efficiency and cell number). That is, the preferred detection method for detecting whether olfactory receptors are activated by compounds in the present invention.

[0093] In the present invention, the term "dose-dependent curve / dose-response relationship" refers to the rule that the intensity of activation of olfactory receptors by functional compounds changes with the change of compound concentration. Usually, the "S" type curve is drawn with the logarithm of compound concentration as the abscissa and the normalized fluorescence signal intensity as the ordinate. This relationship is the key evidence for confirming the specific interaction of "ligand-receptor". That is, the core experimental evidence for confirming the specific biological interaction between compounds and receptors.

[0094] In the present invention, the term "‘compound-olfactory receptor (OR)’ interaction pair" refers to the dose-dependent activation relationship that is proven to exist between a specific functional compound and a specific olfactory receptor. For example, "L-carvone-OR2W1" constitutes a pair.

[0095] In the present invention, the term "new pair" refers to finding and confirming a new specific ligand compound that can activate an olfactory receptor whose ligand is known through experimental methods. That is, the important discovery of the present invention refers to the first reported pair relationship between a compound and an olfactory receptor (OR).

[0096] In the present application, the term "deorphanization" refers to the process of finding and confirming, through experimental methods, a specific ligand compound that can activate an "orphan" olfactory receptor (Orphan olfactory receptors) whose natural ligand is unknown. That is, the breakthrough contribution of the present application at the basic research level refers to the first discovery of a ligand for a receptor (such as OR14I1).

[0097] In the present application, the term "fingerprint / standard fingerprint" refers to a characteristic pattern based on biological response. It refers to a characteristic set unique to a specific natural plant sample, which is composed of all or part of the "functional compound-OR" interaction pairs. The graph can further contain the activation intensity information of each pair. That is, the present application is used to realize the information basis and comparison standard for identification and tracing of natural plant samples.

[0098] The present application proposes the use of olfactory receptors in identifying a predetermined compound in a sample, a recombinant cell, a kit for identifying a predetermined compound in a sample, a set of olfactory receptors for identifying plant varieties, and a method for identifying plant varieties, which will be described in detail below.

[0099] Use of olfactory receptors in identifying a predetermined compound in a sample In one aspect of the present application, the present application proposes the use of olfactory receptors in identifying a predetermined compound in a sample. According to embodiments of the present application, the olfactory receptors include at least one selected from the group consisting of OR14I1, OR2T6, OR5H15, and OR2F2; and the predetermined compound includes at least one selected from the group consisting of isobornyl formate, levorotatory borneol, 1-butyl-2-isobutyl phthalate, 2,3-dihydro-2-methylbenzofuran, citral, diisobutyl phthalate, eugenyl acetate, and citronellyl formate. The use according to the embodiments of the present application has the remarkable effect of successfully applying a newly discovered and specific "compound-olfactory receptor" pair to construct a new natural plant identification method for the first time.

[0100] Specifically, the core contribution of the present application is to complete the "de-orphaning" and functional definition of key olfactory receptors such as OR14I1, OR2T6, OR5H15, OR2F2, and systematically construct their corresponding recognition relationship with a series of plant-derived volatile compounds such as formic acid isobornyl ester and eugenol acetate. Based on this, we can convert complex phytochemical composition information into specific olfactory receptor-mediated biological activation signals that can be qualitative and quantitative. This conversion brings fundamental technical advantages: natural plant samples of different sources will activate a group of characteristic olfactory receptors due to the differences in their volatile component composition, thereby forming their unique "biological activation fingerprint". By analyzing whether the sample to be tested can activate core receptor combinations such as OR2T6 / OR5H15 (indicating eugenol acetate) or OR14I1 (indicating diisobutyl phthalate), and comparing its activation spectrum with the standard spectrum, the identification of plant species, authenticity or origin can be achieved. This functional biological recognition-based strategy is based on the highly specific molecular interaction between receptors and ligands, so compared with simple chemical composition analysis, it has higher intrinsic specificity and anti-matrix interference ability, providing a more reliable new way for the accurate identification of natural plants.

[0101] According to embodiments of the present application, the predetermined compound includes at least one selected from the group consisting of diisobutyl phthalate and eugenol acetate.

[0102] According to embodiments of the present application, the olfactory receptor further includes at least one of OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR12D2, OR5T2, OR4D2, OR51A7, and OR2J3.

[0103] According to embodiments of the present application, the OR2W1 is used to recognize at least one of the isobornyl formate and the levoborneol; or the OR2C1 is used to recognize the levoborneol; or the OR2M3 is used to recognize the levoborneol; or the OR2T11 is used to recognize the levoborneol; or the OR10X1 is used to recognize the levoborneol; or the OR52L1 is used to recognize the levoborneol; or the OR7A17 is used to recognize the levoborneol; or the OR9I1 is used to recognize the phthalic acid-1-butyl ester-2-isobutyl ester; or the OR5K1 is used to recognize the 2,3-dihydro-2-methylbenzofuran; or the OR3A3 is used to recognize at least one of the citral and the diisobutyl phthalate; or the OR7E24 is used to recognize the citral; or the OR14A2 is used to recognize the diisobutyl phthalate; or the OR14I1 is used to recognize the diisobutyl phthalate; or the OR12D2 is used to recognize the diisobutyl phthalate; or the OR5T2 is used to recognize the diisobutyl phthalate; or the OR4D2 is used to recognize the diisobutyl phthalate; or the OR2T6 is used to recognize the eugenyl acetate; or the OR5H15 is used to recognize the eugenyl acetate; or the OR2F2 is used to recognize the eugenyl acetate; or the OR51A7 is used to recognize the eugenyl acetate; or the OR2J3 is used to recognize the citronellyl formate.

[0104] According to embodiments of the present application, the olfactory receptor has an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, and SEQ ID NO: 22.

[0105] According to embodiments of the present application, the OR14I1 has an amino acid sequence as set forth in SEQ ID NO: 14.

[0106] According to embodiments of the present application, the OR2T6 has an amino acid sequence as set forth in SEQ ID NO: 18.

[0107] According to embodiments of the present application, the OR5H15 has an amino acid sequence as set forth in SEQ ID NO: 19.

[0108] According to embodiments of the present application, the OR2F2 has an amino acid sequence as set forth in SEQ ID NO: 21.

[0109] According to embodiments of the present application, the OR2W1 has an amino acid sequence as set forth in SEQ ID NO: 1.

[0110] According to embodiments of the present application, the OR2C1 has an amino acid sequence as set forth in SEQ ID NO: 2.

[0111] According to embodiments of the present application, the OR2M3 has an amino acid sequence as set forth in SEQ ID NO: 3.

[0112] According to embodiments of the present application, the OR2T11 has an amino acid sequence as set forth in SEQ ID NO: 4.

[0113] According to embodiments of the present application, the OR10X1 has an amino acid sequence as set forth in SEQ ID NO: 5.

[0114] According to embodiments of the present application, the OR52L1 has an amino acid sequence as set forth in SEQ ID NO: 6.

[0115] According to embodiments of the present application, the OR7A17 has an amino acid sequence as set forth in SEQ ID NO: 7.

[0116] According to embodiments of the present application, the OR9I1 has an amino acid sequence as set forth in SEQ ID NO: 8.

[0117] According to embodiments of the present application, the OR5K1 has an amino acid sequence as set forth in SEQ ID NO: 9.

[0118] According to embodiments of the present application, the OR3A3 has an amino acid sequence as set forth in SEQ ID NO: 11.

[0119] According to embodiments of the present application, the OR7E24 has an amino acid sequence as set forth in SEQ ID NO: 12.

[0120] According to embodiments of the present application, the OR14A2 has an amino acid sequence as set forth in SEQ ID NO: 13.

[0121] According to embodiments of the present application, the OR12D2 has an amino acid sequence as set forth in SEQ ID NO: 15.

[0122] According to embodiments of the present application, the OR5T2 has an amino acid sequence as shown in SEQ ID NO: 16.

[0123] According to embodiments of the present application, the OR4D2 has an amino acid sequence as shown in SEQ ID NO: 17.

[0124] According to embodiments of the present application, the OR51A7 has an amino acid sequence as shown in SEQ ID NO: 22.

[0125] According to embodiments of the present application, the OR2J3 has an amino acid sequence as shown in SEQ ID NO: 20.

[0126] According to embodiments of the present application, the sample is a plant sample.

[0127] Recombinant cell In a second aspect of the present application, the present application provides a recombinant cell. According to embodiments of the present application, the recombinant cell expresses an olfactory receptor and a reporter protein; the olfactory receptor comprises at least one selected from the group consisting of OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7 and OR2J3; the reporter protein is suitable for generating a detectable signal after the olfactory receptor recognizes the predetermined compound. The recombinant cell according to embodiments of the present application successfully constructs a high-efficiency, standardized biosensor core element by combining the newly discovered olfactory receptor of the present application with a reporter protein system; this design directly converts the specific recognition behavior of specific odor molecules and receptors into a universal, quantifiable optical or electrochemical signal, thereby simplifying the complex chemical detection problem into an intuitive signal reading process, not only providing a stable, reusable detection tool for precise identification of natural plants, but also laying a solid foundation for realizing high-throughput, automated odor fingerprint analysis.

[0128] According to embodiments of the present application, the olfactory receptor is expressed on the cell membrane of the recombinant cell.

[0129] Kit for identifying a predetermined compound in a sample In a third aspect, the present application provides a kit for identifying a predetermined compound in a sample. According to embodiments of the present application, the kit comprises the recombinant cell of the second aspect of the present application. According to the kit of embodiments of the present application, the detection capability of the recombinant cell can be standardized and productized, providing a complete solution ready for use. The kit combines the recombinant cells expressing different olfactory receptors in a reasonable manner or is packaged separately, and is equipped with necessary detection reagents, so that even non-professionals can quickly and accurately complete the synchronous detection and identification of multiple characteristic odor molecules in complex samples under routine experimental conditions, greatly improving the operability and repeatability of the olfactory receptor-based detection method, and providing key support for the practical popularization and application of natural plant identification technology.

[0130] A set of olfactory receptors for identifying plant varieties In a fourth aspect, the present application provides a set of olfactory receptors for identifying plant varieties. According to embodiments of the present application, the olfactory receptors comprise at least one selected from OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7 and OR2J3. According to the olfactory receptors of embodiments of the present application, the core effect is to provide a set of functionally verified and special receptor combinations that can be used to build a plant variety identification system. Each member of the receptor set has been proven to be able to specifically respond to one or more plant characteristic volatile compounds, such as the key pair of OR2T6 and eugenyl acetate, OR14I1 and diisobutyl phthalate. When these receptors are used as a whole, they are no longer just isolated detection targets, but together constitute the core elements of a "biological sensor array" that can decode complex plant odor fingerprints. By selectively combining different members of the receptor set, the multi-dimensional biological response signal of the plant to be tested can be systematically collected, thereby establishing a unique identification model that can accurately distinguish different plant varieties. The establishment of this set of receptors provides the most fundamental and optimized material basis for the development of various olfactory receptor-based identification products (including but not limited to recombinant cells, kits and detection methods), ensuring the high specificity and reliability of the final identification results.

[0131] A method for identifying plant varieties In a fifth aspect, the present application provides a method for identifying a plant variety. According to embodiments of the present application, the method comprises detecting a plant sample using the recombinant cell of the second aspect, the kit of the third aspect, or the olfactory receptor of the fourth aspect of the present application; and determining the variety of the plant sample based on the detection result.

[0132] According to embodiments of the present application, the method comprises stimulating the olfactory receptor in the recombinant cell of the second aspect, the kit of the third aspect, or the olfactory receptor of the fourth aspect of the present application with a volatile component of a plant variety to be tested, obtaining a response value; determining whether the volatile component of the plant variety to be tested can activate the olfactory receptor based on the response value; determining the volatile component of the plant variety to be tested based on the category of the activated receptor; and determining the variety of the plant to be tested based on the category of the volatile component of the plant variety to be tested.

[0133] According to embodiments of the present application, the response value being not lower than a preset threshold value indicates that the volatile component of the plant variety to be tested can activate the olfactory receptor.

[0134] According to embodiments of the present application, the OR2W1 having an activation signal indicates that the volatile component of the plant variety to be tested has at least one of isobornyl formate and levoborneol.

[0135] According to embodiments of the present application, the OR2C1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0136] According to embodiments of the present application, the OR2M3 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0137] According to embodiments of the present application, the OR2T11 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0138] According to embodiments of the present application, the OR10X1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0139] According to embodiments of the present application, the OR52L1 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0140] According to embodiments of the present application, the OR7A17 having an activation signal indicates that the volatile component of the plant variety to be tested has levoborneol.

[0141] According to embodiments of the application, OR9I1 has an activation signal that is an indication that the volatile components of the plant variety under test have 1-butyl 2-isobutyl phthalate.

[0142] According to embodiments of the application, OR5K1 has an activation signal that is an indication that the volatile components of the plant variety under test have 2,3-dihydro-2-methylbenzofuran.

[0143] According to embodiments of the application, OR3A3 has an activation signal that is an indication that the volatile components of the plant variety under test have at least one of citral and diisobutyl phthalate.

[0144] According to embodiments of the application, OR7E24 has an activation signal that is an indication that the volatile components of the plant variety under test have citral.

[0145] According to embodiments of the application, OR14A2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0146] According to embodiments of the application, OR14I1 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0147] According to embodiments of the application, OR12D2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0148] According to embodiments of the application, OR5T2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0149] According to embodiments of the application, OR4D2 has an activation signal that is an indication that the volatile components of the plant variety under test have diisobutyl phthalate.

[0150] According to embodiments of the application, OR2T6 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0151] According to embodiments of the application, OR5H15 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0152] According to embodiments of the application, OR2F2 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0153] According to embodiments of the application, OR51A7 has an activation signal that is an indication that the volatile components of the plant variety under test have eugenol acetate.

[0154] According to an embodiment of the present application, OR2J3 has an activation signal, and is an indication that the volatile components of the plant variety under test have citronellal formate.

[0155] According to an embodiment of the present application, the volatile components of the plant variety under test have at least one of isobornyl formate and levoborneol, and is an indication that the plant under test is a genus of Dipterocarpaceae.

[0156] According to an embodiment of the present application, the volatile components of the plant variety under test have levoborneol, and is an indication that the plant under test is a genus of Dipterocarpaceae.

[0157] According to an embodiment of the present application, the volatile components of the plant variety under test have 1-butyl 2-isobutyl phthalate, and is an indication that the plant under test is a genus of Syzygium.

[0158] According to an embodiment of the present application, the volatile components of the plant variety under test have 2,3-dihydro-2-methylbenzofuran, and is an indication that the plant under test is a genus of Syzygium.

[0159] According to an embodiment of the present application, the volatile components of the plant variety under test have citral, and is an indication that the plant under test is a genus of Zingiber.

[0160] According to an embodiment of the present application, the volatile components of the plant variety under test have diisobutyl phthalate, and is an indication that the plant under test is a genus of Agastache.

[0161] According to an embodiment of the present application, the volatile components of the plant variety under test have eugenyl acetate, and is an indication that the plant under test is a genus of Myristica.

[0162] According to an embodiment of the present application, the volatile components of the plant variety under test have citronellal formate, and is an indication that the plant under test is a genus of Magnolia.

[0163] The sequence of the present application is shown as follows:

[0164] The scheme of the present application will be explained below in conjunction with examples. Those skilled in the art will understand that the following examples are only for illustrating the present application, and should not be regarded as limiting the scope of the present application. If a specific technique or condition is not mentioned in the examples, the technique or condition described in the literature in the art or according to the product manual is used. If the manufacturer of the reagent or instrument is not mentioned, it is a conventional product that can be obtained in the market.

[0165] Example 1: Construction of a library of biomimetic olfactory receptor cells (1) Cell culture: Human embryonic kidney cells HEK293T were cultured in a 37°C, 5% CO2 incubator using DMEM high glucose medium containing 10% fetal bovine serum.

[0166] (2) Plasmid preparation: Approximately 400 human olfactory receptor (OR) coding sequences published in the NCBI database were selected, synthesized, and cloned into an expression vector. Meanwhile, plasmids expressing Golf protein, CRE-Luciferase reporter gene, and pRL-SV40 internal standard were prepared.

[0167] (3) Co-transfection: When the density of HEK293T cells in the microplate reached 80-90%, gene constructs containing olfactory receptors, Golf, CRE-Luciferase, and pRL-SV40 were prepared, and the gene constructs were co-transfected into HEK293T cells using liposome transfection reagents.

[0168] (4) Cell library preparation: 24 hours after transfection, the cells can be used for high-throughput screening. To maintain consistency, all screening experiments were completed within 24-48 hours after transfection.

[0169] Example 2: Analysis of volatile components of natural plant samples and screening of key compounds (1) Sample pretreatment: Six different natural plant samples (numbered Sample 1 to Sample 6) were ground into powder. 0.1 g of powder was accurately weighed into a 20 mL headspace sampling bottle.

[0170] (2) SPME extraction: The aged 50 / 30 μm DVB / CAR / PDMS extraction head was inserted into the headspace bottle, and headspace extraction was performed at 60°C for 40 minutes.

[0171] (3) GC-MS analysis: After extraction was completed, the extraction head was inserted into the GC sampling port, desorbed at 260°C for 5 minutes, and subjected to GC-MS analysis.

[0172] (a) Chromatographic conditions: DB-5MS chromatographic column (30 m x 0.25 mm x 0.25 μm); programmed temperature: initial 50°C for 3 minutes, increased to 120°C at 15°C / min, then increased to 205°C at 3°C / min, and then increased to 260°C at 10°C / min, and held for 7 minutes.

[0173] (b) Mass spectrometry conditions: Electron impact ion source (EI), ion source temperature 230°C, electron energy 70 eV, scan range m / z 20-450.

[0174] (4) Data processing: Qualitative analysis of each chromatographic peak was performed by NIST 20 version standard mass spectral library, and the relative percentage content of each compound was calculated by peak area normalization method. The results are shown in Table 1 and Figure 1 , Figure 2 .

[0175] (5) Key compound screening: From each sample, the compound with a relative percentage content greater than 0.5% and easy to purchase was selected as a key compound for subsequent receptor screening. For example, isobornyl formate and levoborneol were selected from sample 1.

[0176] Table 1: Main volatile odor compounds in 6 natural plant samples

[0177] Example 3: High-throughput screening and verification of olfactory receptor action target Taking levoborneol in sample 1 as an example, the screening and verification process is shown: (1) Preliminary screening: Levoborneol was prepared into a 1 M stock solution with DMSO, and then diluted into a 3 mM working concentration with a detection buffer. 25 μL of the working solution was added to the microplate with the constructed cell library, 2 replicates were set, and a well containing only the buffer was set as a negative control. After 4 hours of stimulation, the Dual-Glo TM Luciferase Assay System kit was used to detect the luminescence signals of firefly luciferase and sea pansy luciferase in turn on a multifunctional enzyme label instrument. The sea pansy luciferase signal was used as an internal reference for normalization.

[0178] (2) Result interpretation: The relative fluorescence units after normalization were calculated. Compared with the negative control group, the wells with significantly increased signal values (for example, fold change > 2 and p < 0.05) were determined as positive wells. By the OR species located in the well, it can be preliminarily determined that levoborneol activates the OR (for example, OR2W1).

[0179] (3) Dose-dependent verification: For the positive pair determined by preliminary screening (levoborneol vs OR2W1), a dose-dependent experiment was performed. Levoborneol was gradiently diluted in a series, and the cells expressing OR2W1 were stimulated respectively. Taking the logarithm of the compound concentration as the horizontal coordinate and the normalized fluorescence signal value as the vertical coordinate, the dose-response curve was drawn by fitting with the four-parameter logistic equation, confirming its typical dose-dependent nature. The results are shown in Figure 3 .

[0180] For other samples (Sample 2 to Sample 6), the same dose-dependent experiment was also performed in this embodiment to verify the interaction of each functional compound with the specific olfactory receptor. The dose-response curves are shown in Figure 4 (Sample 2), Figure 5 (Sample 3), Figure 6 (Sample 4), Figure 7 , Figure 8 (Sample 5), and Figure 9 (Sample 6). All curves show a typical dose-dependent, further confirming the specificity and strength of the compound-receptor pair.

[0181] Through the above process, all key compounds in the 6 samples were systematically screened in this embodiment, and finally 34 "compound-OR" interaction pairs were identified, as shown in Table 2. Among them, 23 are new pairs, as shown in Table 3, and 4 ORs are first "orphaned", as shown in Table 4.

[0182] Table 2: Summary of olfactory receptor target points of functional compounds in 6 natural plant samples

[0183] Table 3: Summary of new compound-olfactory receptor (OR) pairs

[0184] Table 4: Summary of first "orphaned" OR and its ligand compound

[0185] Example 4: Identification application of natural plant samples (1) Establish a standard fingerprint: based on the results of Examples 2 and 3, a standard fingerprint is established for Sample 5. This fingerprint is defined as the set of 11 ORs (OR2T6, OR5H15, OR2J3, OR2F2, OR51A7, OR6T1, OR10G4, OR10G7, OR2W1, OR10A5, OR52A4) that can be specifically activated by the functional compound eugenol acetate in Sample 5.

[0186] (2) Identification of the sample to be tested: a natural plant sample to be tested is obtained, named Sample 5. According to the same method of Examples 2 and 3, the volatile components are analyzed, and the activation of the above 11 ORs by the extract or the main component (such as eugenol acetate) thereof is tested.

[0187] (3) Result comparison and identification: (a) True product determination: if the sample to be tested can activate more than a preset threshold (for example, 9 or more) of the ORs in the standard fingerprint map, and the characteristics of the dose curve of the sample to be tested are consistent with the standard fingerprint map, the sample to be tested is determined as a true product sample 5.

[0188] (b) Fake product determination: if the sample to be tested can only activate a small number of ORs in the standard fingerprint map or cannot activate any OR, the sample to be tested is determined as a fake or inferior product.

[0189] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in the present specification and the features of the different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0190] Although the embodiments of the present application have been shown and described above, it is understood that the above-described embodiments are exemplary and are not to be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above-described embodiments within the scope of the present application.

Claims

1. The use of olfactory receptors in identifying predetermined compounds in a sample, characterized in that, The olfactory receptors include at least one selected from the following: OR14I1, OR2T6, OR5H15, OR2F2; the predetermined compound includes at least one selected from the following: isoborneol formate, levoborneol, 1-butyl-2-isobutyl phthalate, 2,3-dihydro-2-methylbenzofuran, citral, diisobutyl phthalate, eugenol acetate, citronellol formate.

2. The use according to claim 1, characterized in that, The predetermined compound includes at least one of the following: diisobutyl phthalate, eugenol acetate.

3. The use according to claim 1 or 2, characterized in that, The olfactory receptors further include at least one of the following: OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR12D2, OR5T2, OR4D2, OR51A7, and OR2J3.

4. The use according to any one of claims 1 to 3, characterized in that, The OR2W1 is used to identify at least one of isoborneol formate and levoborneol; or The OR2C1 is used to identify the left-handed borneol; or The OR2M3 is used to identify the left-handed borneol; or The OR2T11 is used to identify the left-handed borneol; or The OR10X1 is used to identify the left-handed borneol; or The OR52L1 is used to identify the left-handed borneol; or The OR7A17 is used to identify the left-handed borneol; or OR9I1 is used to identify 1-butyl phthalate-2-isobutyl phthalate; or The OR5K1 is used to identify the 2,3-dihydro-2-methylbenzofuran; or The OR3A3 is used to identify at least one of citral and diisobutyl phthalate; or OR7E24 is used to identify citral; or OR14A2 is used to identify the diisobutyl phthalate; or The OR14I1 is used to identify the diisobutyl phthalate; or OR12D2 is used to identify diisobutyl phthalate; or The OR5T2 is used to identify the diisobutyl phthalate; or The OR4D2 is used to identify the diisobutyl phthalate; or OR2T6 is used to identify the eugenol acetate; or OR5H15 is used to identify the eugenol acetate; or The OR2F2 is used to identify the eugenol acetate; or OR51A7 is used to identify the eugenol acetate; or OR2J3 is used to identify the citronellol formate.

5. The use according to claim 3, characterized in that, The olfactory receptor has an amino acid sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21 or SEQ ID NO:22; Optionally, the OR14I1 has an amino acid sequence as shown in SEQ ID NO:14; Optionally, the OR2T6 has the amino acid sequence shown in SEQ ID NO:18; Optionally, OR5H15 has an amino acid sequence as shown in SEQ ID NO:19; Optionally, the OR2F2 has an amino acid sequence as shown in SEQ ID NO:21; Optionally, the OR2W1 has an amino acid sequence as shown in SEQ ID NO:1; Optionally, the OR2C1 has an amino acid sequence as shown in SEQ ID NO:2; Optionally, the OR2M3 has an amino acid sequence as shown in SEQ ID NO:3; Optionally, the OR2T11 has an amino acid sequence as shown in SEQ ID NO:4; Optionally, the OR10X1 has an amino acid sequence as shown in SEQ ID NO:5; Optionally, the OR52L1 has an amino acid sequence as shown in SEQ ID NO:6; Optionally, the OR7A17 has the amino acid sequence shown in SEQ ID NO:7; Optionally, the OR9I1 has the amino acid sequence shown in SEQ ID NO:8; Optionally, the OR5K1 has an amino acid sequence as shown in SEQ ID NO:9; Optionally, the OR3A3 has an amino acid sequence as shown in SEQ ID NO:11; Optionally, OR7E24 has an amino acid sequence as shown in SEQ ID NO:12; Optionally, the OR14A2 has an amino acid sequence as shown in SEQ ID NO:13; Optionally, the OR12D2 has an amino acid sequence as shown in SEQ ID NO:15; Optionally, the OR5T2 has an amino acid sequence as shown in SEQ ID NO:16; Optionally, the OR4D2 has an amino acid sequence as shown in SEQ ID NO:17; Optionally, the OR51A7 has an amino acid sequence as shown in SEQ ID NO:22; Optionally, the OR2J3 has an amino acid sequence as shown in SEQ ID NO:

20.

6. The use according to any one of claims 1-5, characterized in that, The sample is a plant sample.

7. A recombinant cell, characterized in that, The recombinant cells express olfactory receptors and reporter proteins; The olfactory receptors include at least one of the following: OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7 and OR2J3; The reporter protein is adapted to generate a detectable signal after the olfactory receptor recognizes the predetermined compound.

8. The recombinant cell according to claim 7, characterized in that, The olfactory receptors are expressed on the cell membrane of the recombinant cells.

9. A kit for identifying a predetermined compound in a sample, characterized in that, It comprises: at least one recombinant cell as described in claim 7 or 8.

10. A group of olfactory receptors for identifying plant varieties, characterized in that, The olfactory receptors include at least one selected from OR2W1, OR2C1, OR2M3, OR2T11, OR10X1, OR52L1, OR7A17, OR9I1, OR5K1, OR3A3, OR7E24, OR14A2, OR14I1, OR12D2, OR5T2, OR4D2, OR2T6, OR5H15, OR2F2, OR51A7, and OR2J3.

11. A method for identifying plant varieties, characterized in that, include: Detection of plant samples using the recombinant cells according to any one of claims 7-8, the kit according to claim 9, or the olfactory receptor according to claim 10; Based on the test results, the species of the plant sample was determined.

12. The method according to claim 11, characterized in that, The method includes: The olfactory receptors in the recombinant cells of any one of claims 7-8, the kit of claim 9, or the olfactory receptors of claim 10 are stimulated by the volatile components of the plant variety to be tested to obtain a response value; Based on the response value, it is determined whether the volatile component stimulation of the tested plant variety can activate the olfactory receptor. Based on the type of activating receptor, the volatile components of the tested plant variety were determined; The variety of the plant to be tested is determined based on the types of volatile components of the plant to be tested. Optionally, the response value not being lower than a preset threshold is an indication that the volatile components of the plant variety being tested can activate the olfactory receptors; Optionally, OR2W1 has an activation signal that indicates that the volatile components of the tested plant variety contain at least one of isoborneol formate and L-borneol. Optionally, OR2C1 has an activation signal that indicates the presence of levorotatory borneol in the volatile components of the tested plant species; Optionally, OR2M3 has an activation signal that indicates the presence of levorotatory borneol in the volatile components of the tested plant species; Optionally, OR2T11 has an activation signal that indicates the presence of levorotatory borneol in the volatile components of the tested plant species; Optionally, OR10X1 has an activation signal that indicates the presence of levorotatory borneol in the volatile components of the tested plant species. Optionally, OR52L1 has an activation signal that indicates the presence of levorotatory borneol in the volatile components of the tested plant species. Optionally, OR7A17 has an activation signal and is an indicator of the presence of levorotatory borneol in the volatile components of the tested plant species; Optionally, OR9I1 has an activation signal and is an indicator of the volatile components of the tested plant species containing 1-butyl phthalate-2-isobutyl phthalate. Optionally, OR5K1 has an activation signal that indicates the presence of 2,3-dihydro-2-methylbenzofuran in the volatile components of the tested plant species; Optionally, OR3A3 has an activation signal that indicates the presence of at least one of citral and diisobutyl phthalate in the volatile components of the tested plant variety. Optionally, OR7E24 has an activation signal and is an indicator of the presence of citral in the volatile components of the tested plant species; Optionally, OR14A2 has an activation signal that indicates the presence of diisobutyl phthalate in the volatile components of the tested plant species. Optionally, OR14I1 has an activation signal and is an indicator of diisobutyl phthalate in the volatile components of the tested plant species. Optionally, OR12D2 has an activation signal and is an indicator of diisobutyl phthalate in the volatile components of the tested plant species. Optionally, OR5T2 has an activation signal that indicates the presence of diisobutyl phthalate in the volatile components of the tested plant species. Optionally, OR4D2 has an activation signal that indicates the presence of diisobutyl phthalate in the volatile components of the tested plant species. Optionally, OR2T6 has an activation signal and is an indicator of the presence of eugenol acetate in the volatile components of the tested plant species; Optionally, OR5H15 has an activation signal and is an indicator of the presence of eugenol acetate in the volatile components of the tested plant species. Optionally, OR2F2 has an activation signal and is an indicator of the presence of eugenol acetate in the volatile components of the tested plant species; Optionally, OR51A7 has an activation signal and is an indicator of the presence of eugenol acetate in the volatile components of the tested plant species; Optionally, OR2J3 has an activation signal and is an indicator of the presence of citronellol formate in the volatile components of the tested plant species. Optionally, the presence of at least one of isoborneol formate and L-borneol in the volatile components of the plant species to be tested is an indication that the plant species to be tested is a plant of the genus Dipterocarpus. Optionally, the presence of levorotatory borneol in the volatile components of the plant species to be tested is an indication that the plant species to be tested is a plant of the genus Dipterocarpus. Optionally, the volatile components of the plant variety to be tested contain 1-butyl phthalate-2-isobutyl phthalate, which is an indicator that the plant to be tested is a styrax plant. Optionally, the presence of 2,3-dihydro-2-methylbenzofuran in the volatile components of the plant species being tested is an indication that the plant being tested is a species of the genus Syringa. Optionally, the presence of citral in the volatile components of the plant species being tested is an indicator that the plant is a ginger species. Optionally, the presence of diisobutyl phthalate in the volatile components of the plant species to be tested is an indicator that the plant species to be tested is a member of the genus Patchouli. Optionally, the volatile components of the plant variety to be tested contain eugenol acetate, which is an indicator that the plant to be tested is a Myristica genus. Optionally, the volatile components of the plant species to be tested contain citronellol formate, which is an indicator that the plant species to be tested is a Magnolia species.

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