Methods for identifying modulators of insect transient receptor potential channels - Patents.com

Abstract

The present invention relates to a screening method for determining whether a candidate compound is a modulator of a complex consisting of the insect transient receptor potential channel Nanchung protein and the insect transient receptor potential channel Water witch protein. The present invention further relates to a screening method for determining whether a candidate compound directly binds to a complex consisting of the Nanchung protein and the Water witch protein. The present invention further relates to a screening method for determining whether a candidate compound is a modulator of the insect Water witch protein. The present invention further provides a method for insect control by applying an insect-specific TRP channel modulator determined by the screening method. The present invention further provides a method for classifying insecticide compounds by their mechanism of action using binding assays and / or functional assays.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 952,756, filed December 23, 2019, which is incorporated herein by reference in its entirety.

[0002] Submission of sequence listing The sequence listing associated with this application has been submitted electronically via EFS-Web and is incorporated herein by reference in its entirety. The text file containing the sequence listing is titled "SEQ LISTING 180852WO01." The text file size is 702 KB and the text file was created on December 8, 2020. [Background technology]

[0003] The present invention relates to compositions and methods for insect control. The present invention generally relates to methods for determining agents that directly bind to and / or modulate the biological activity of insect transient receptor potential A (TRPA) Water witch protein and / or a complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential A (TRPA) Water witch protein. The present invention also relates to compositions and methods of a mode-of-action class of insecticide compositions comprising compounds that can modulate the biological activity of intracellular insect transient receptor potential A (TRPA) Water witch protein and / or a complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential A (TRPA) Water witch protein. The present invention further relates to compositions and methods of a mode-of-action class of insecticide compositions comprising compounds that can directly bind to a complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential A (TRPA) Water witch protein.

[0004] Nearly all field crops, plants, and commercial agricultural areas are susceptible to attack by plant insect pests. Plant insect pests are a major cause of losses of commercially important agricultural crops worldwide, resulting in economic hardship for farmers and nutritional deficiencies for people living in many parts of the world. In other words, insect pests cause significant losses and damage to human agriculture, food supply, post-harvest storage, horticulture, animal health, and public health.

[0005] Although advances have been made in the control of these insects, they are able to adopt and evade control measures, resulting in resistant insect populations (Perry et al., 2011). Consequently, there is still a need to better understand the mechanisms underlying the feeding and other behaviors of insect pests. Such knowledge would enable the design of agents and strategies to disrupt or prevent attacks by insect pests on field crops, plants, and commercial agricultural areas, or to prevent the spread of human and animal diseases.

[0006] A wide range of chemical pesticides and insecticides have been widely used to control or eradicate insect pests. However, there is great interest in identifying and / or developing effective alternative insecticides. Therefore, an effective method for screening new insecticides would be desirable.

[0007] Several species or arthropods, including mosquitoes, flies, ticks, small insects, and fleeces, transmit various pathogenic viruses and parasites. Widely used insect repellents, such as DEET, picaridin, and permethrin, have raised several concerns regarding environmental and health risks (Lee, 2018). Therefore, there is great interest in developing new methods to screen for efficient insect repellents with potentially low toxicity to health and the environment.

[0008] Therefore, what is needed are new methods that can be used to screen for agents that can bind to and / or modulate the biological activity of insect transient receptor potential A (TRPA) Water witch proteins and / or the complex consisting of insect transient receptor potential V (TRPV) Nanchung proteins and insect transient receptor potential A (TRPA) Water witch proteins, and potentially modulate insect behavior and act as insecticides and / or insect repellents. Summary of the Invention

[0009] The present invention relates to methods for determining the Insecticide Resistance Action Committee (IRAC) mode of action classification of a candidate compound using one or more functional and / or binding assays.

[0010] For example, in certain embodiments, functional assays may be used to determine whether a candidate compound acts as a modulator of insect transient receptor potential A (TRPA) Water witch protein and / or a complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential A (TRPA) Water witch protein within a cell.

[0011] In yet another embodiment, a functional assay may be used to determine whether a candidate compound acts like an endogenous modulator of the complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential V (TRPV) Inactive protein in cells.

[0012] In yet another embodiment, functional assays may be used to determine whether a candidate compound acts like an endogenous modulator of the complex consisting of insect transient receptor potential V (TRPV) Nanchung protein and insect transient receptor potential A (TRPA) Water witch protein within cells.

[0013] The functional assay may include at least one of the following steps: (1) detecting calcium ion mobilization in the first cell in response to a candidate compound or an endogenous modulator; (2) detecting membrane potential in the first cell in response to the candidate compound on the endogenous modulator; (3) comparing calcium ion mobilization in the first cell in the absence of the candidate compound or endogenous modulator with calcium ion mobilization in the first cell in the presence of the candidate compound or endogenous modulator; and (4) comparing the membrane potential in the first cell in the absence of the candidate compound or endogenous modulator with the membrane potential in the first cell in the presence of the candidate compound or endogenous modulator. (5) comparing calcium ion mobilization in the first cell in the presence of the candidate compound or endogenous modulator with a reference level of calcium ion mobilization indicating no modulation of the TPRV channel and / or TRPA channel and / or the complex consisting of insect transient receptor potential V (TRPV) channel and insect transient receptor potential A (TRPA) channel; (6) comparing membrane potential in the first cell in the presence of the candidate compound or endogenous modulator with a reference level of membrane potential indicating no modulation of the TPRV channel and / or TRPA channel and / or the complex consisting of insect transient receptor potential V (TRPV) channel and insect transient receptor potential A (TRPA) channel. Preferably, the candidate compound or endogenous modulator can modulate calcium ion mobilization or membrane potential in the first cell by at least 20% relative to the reference level. The candidate compound or endogenous modulator can be a modulator that inhibits the activity of insect TRPV channel and / or TRPA channel and / or the complex consisting of insect transient receptor potential V (TRPV) channel and insect transient receptor potential A (TRPA) channel. Alternatively, the candidate compound or endogenous modulator may be a modulator that activates the activity of an insect TRPV channel and / or an insect TRPA channel and / or a complex consisting of an insect transient receptor potential V (TRPV) channel and an insect transient receptor potential A (TRPA) channel.Candidate compounds may be modulators that kill insects and / or inhibit insect feeding and / or interfere with insect behavior.

[0014] In another embodiment, a binning assay may be used to determine whether a candidate compound binds directly to a complex consisting of transient receptor potential (TRPV) proteins and transient receptor potential A (TRPA) proteins.

[0015] In another embodiment, binding assays can be used to determine whether a candidate compound binds to the same site as an endogenous modulator of the complex consisting of insect transient receptor potential V (TRPV) protein and insect transient receptor potential A (TRPA) protein.

[0016] In another embodiment, binding assays can be used to determine whether a candidate compound binds to the same site as an endogenous modulator of a complex or channel consisting of transient receptor potential (TRPV) proteins and transient receptor potential A (TRPA) proteins.

[0017] In another embodiment, binding assays can be used to determine whether two or more candidate compounds bind to the same or different sites on the complex formed by transient receptor potential (TRPV) proteins and transient receptor potential A (TRPA) proteins.

[0018] The binding assay may include: (1) providing one or more Nanchung proteins, transient receptor potential V (TRPV) inactive proteins, and / or transient receptor potential A (TRPA) Water witch proteins, which may be located within, derived from, or produced by a cell, or produced in a cell-free system; (2) contacting one or more TRPV Nanchung proteins and / or TRPV inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of an insect transient receptor potential V (TRPV) Nanchung protein and an insect transient receptor potential A (TRPA) Water witch protein; and the binding affinity of the candidate compound indicates the ability of the candidate compound to directly bind to one or more Nanchung proteins, TRPV inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of an insect transient receptor potential V (TRPV) Nanchung protein and an insect transient receptor potential A (TRPA) Water witch protein.

[0019] For example, in certain embodiments, the binding assay step may include at least one of the following steps: (1) contacting one or more TRPV Nanchung proteins and / or TRPV Inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPA Water witch proteins, with a compound, where the compound may be labeled or unlabeled; (2) measuring the amount of compound that binds to one or more Nanchung proteins, TRPV Inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPA Water witch proteins; (3) measuring the equilibrium dissociation constant (K d (4) calculating Kd(I) calculating (a) the maximum binding (Bmax) and (b) the concentration of the labeled compound required to reach 50% of the maximum binding, which may be determined using one or more of the following methods: (I) calculating (a) the maximum binding (Bmax) and (b) the concentration of the labeled compound required to reach 50% of the maximum binding, (II) fluorescence polarization assay, (III) surface plasmon resonance, (IV) isothermal calorimetry, (V) Schild analysis; (5) contacting one or more Nanchung proteins and / or TRPV Inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of a TRPV Nanchung protein and a TRPA Water witch protein with a protein compound having a high affinity for one or more Nanchung proteins and / or TRPV Inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of a TRPV Nanchung protein and a TRPA Water witch protein; (6) contacting one or more Nanchung proteins, TRPV Inactive proteins and / or TRPA Water witch proteins, one or more Nanchung proteins, and / or TRPV Inactive proteins and / or TRPA Water witch proteins, and / or a complex consisting of a TRPV Nanchung protein and a TRPA Water witch protein with a protein compound having a high affinity for one or more Nanchung proteins, TRPV Inactive proteins and / or TRPA Water witch proteins, (7) contacting the complex consisting of witch proteins with a candidate compound or endogenous modulator in the presence of the labeled compound of step 1; (8) contacting the complex consisting of one or more Nanchung proteins, TRPV Inactive proteins, and / or TRPA Water witch proteins, one or more Nanchung proteins, and / or TRPV Inactive proteins, and / or TRPA Water witch proteins, and / or TRPV Nanchung proteins and TRPA Waterincreasing the amount of candidate compound or endogenous modulator available to contact the complex consisting of witch protein until the binding of the labeled compound approaches 0%; (8) determining the equilibrium inhibition constant (K) for the competitive assay using one or more of the following methods: (I) (a) determining the amount of bound labeled compound in the absence of candidate compound or endogenous modulator, and (b) determining the concentration of candidate compound or endogenous modulator required to reduce the amount of bound labeled compound by 50%; (II) a fluorescence polarization assay; and / or (III) a Schild analysis. i ) and / or the equilibrium dissociation constant (K d (9) comparing the inhibition constant of the candidate compound or endogenous modulator with other candidate compounds; (10) comparing the inhibition constant of the candidate compound or endogenous modulator with the dissociation constant of the labeled compound; and (11) comparing the inhibition constant of the candidate compound with the inhibition constants of one or more candidate compounds.

[0020] In the methods described herein, one or more insect TRPV channels and / or one or more insect TRPA channels may comprise one or more TRPV proteins and / or one or more TRPA proteins in a complex. In the methods described herein, one or more insect TRPV channels and / or TRPA channels may comprise one or more Nanchung proteins, inactive proteins, water witch proteins, and / or Nanchung and inactive protein complexes, and / or Nanchung and water witch protein complexes. For clarity, TRPV and / or TRPA channels may also comprise Nanchung proteins, inactive proteins, and / or water witch proteins found in cell membranes, inserted into artificial membranes, inserted into Nanchungodisks, solubilized in detergents, solubilized in non-detergent polymers, solubilized in amphiphilic polymers (amphipols), and / or complexes consisting of TRPV Nanchung proteins and TRPA water witch proteins produced in cell-free systems. It is understood that Nanchung, Inactive, and Water witch proteins, and / or complexes consisting of one or more Nanchung proteins, and / or TRPV Inactive proteins, and / or TRPA Water witch proteins, and / or TRPV Nanchung proteins and TRPA Water witch proteins, can be found within cell membranes, in solution, or in cell-free systems.It should be understood that the Nanchung protein, Inactive protein, Water witch protein, and / or complexes consisting of TRPV Nanchung protein and TRPA Water witch protein may be contained within one or more cell membranes, within one or more pieces of cell membrane, produced in a cell system, produced in a cell-free system, and / or from cultured cells in a Petri dish, or from cells or tissues derived from a laboratory animal or laboratory insect. For purposes of determining whether a candidate compound directly binds to an insect TRPV protein and / or TRPA protein, the TRPV and / or TRPA channel may contain one or more Nanchung, Inactive, or Water witch proteins.

[0021] It should be understood that both Nanchung and Inactive Protein are insect TRPV proteins. Water Witch is an insect TRPA protein. Nanchung Protein can form a complex with Inactive Protein and / or Water Witch Protein. Candidate compounds can be organic small molecules, inorganic small molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof. Endogenous modulators can be organic small molecules, inorganic small molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

[0022] In another embodiment, the above method may use one or more of the following compounds as the labeling compound: afidopiropene, pymetrozine, and / or the N-deacetylated form of pyrifluquinazone.

[0023] In yet another embodiment, the present invention relates to a candidate compound selected by the above method. The candidate compound may be a modulator that inhibits or increases the activity of an insect TRPV channel or an insect TRPA channel. The candidate compound may act as a pesticide and / or inhibit the feeding behavior of insects. Furthermore, the candidate compound may regulate and / or directly bind to one or more TRPV Nanchung proteins, and / or TRPV Inactive proteins, and / or TRPA proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPV Inactive proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPA Water Witch proteins. The candidate compound may further regulate and / or directly bind to the same site on one or more insect Nanchung TRPV proteins, and / or insect TRPV Inactive proteins, and / or insect TRPA Water witch proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPV Inactive proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPA Water witch proteins, in particular an insect Nanchung TRPV protein, and / or insect TRPV Inactive proteins, and / or insect TRPA Water witch proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPV Inactive proteins, and / or a complex consisting of TRPV Nanchung proteins and TRPA Water witch proteins.Candidate compounds may also regulate and / or directly bind to one or more arthropod Nanchung TRPV proteins, and / or arthropod TRPV Inactive proteins, and / or arthropod TRPA Water witch proteins, and / or complexes consisting of arthropod TRPV Nanchung proteins and arthropod TRPV Inactive proteins, and / or complexes consisting of arthropod TRPV Nanchung proteins and arthropod TRPA Water witch proteins. Candidate compounds may also directly regulate and / or bind to one or more nematode Nanchung TRPV proteins, and / or nematode TRPV Inactive proteins, and / or nematode TRPA Water witch proteins, and / or complexes consisting of nematode TRPV Nanchung proteins and nematode TRPV Inactive proteins, and / or complexes consisting of nematode TRPV Nanchung proteins and nematode TRPA Water witch proteins, and / or other non-target / non-pest insect species.

[0024] In another embodiment, the invention relates to a composition containing a candidate compound identified by one or more of the methods described herein.

[0025] In another embodiment, the present invention relates to a method of insect control, comprising applying a composition containing a candidate compound selected by one or more of the methods described herein, or a compound selected by one or more of the above methods, to an insect, or to an insect habitat, feeding source, and / or environment / habitat. The compound may be an inhibitor of insect TRPV and / or TRPA channels. The compound may be an activator of insect TRPV and / or TRPA channels. The compound may directly bind to insect Nanchung protein or Water witch protein. The insect may be an agricultural / horticultural pest, a disease vector, or a parasite. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 shows a schematic diagram of an expression cassette for an insect TRP channel in an adenoviral expression vector. [Figure 2A] Figures 2A and 2B show the dose-response relationship of calcium responses induced by afidopiropene (AP) and pymetrozine (PM) in cells co-expressing Water witch and Nanchung (Wtrw+Nan), but not by flonicamide (FL) or TFNA-AM, and not in cells expressing Water witch alone (Wtrw). [Figure 2B] Continued from Figure 2A. [Figure 3] Figure 1 shows the physical association of Nanchung (Nan-HA) with Water witch (Wtrw-FLAG) protein in a co-immunoprecipitation experiment. [Figure 4A] Activation of Ca2+ mobilization as a function of time by the endogenous agonist nicotinamide in mammalian cells co-expressing Nanchung and Inactive (Nan+Iav) or Nanchung and Water witch (Nan+Wtrw) (Ca2+ mobilization is not activated in parental mammalian cells (cells before transfection with the gene or protein of interest), cells co-expressing Inactive and Water witch (Iav+Wtrw), or cells expressing Nanchung (Nan), Inactive (Iav), or Water witch (Wtrw) individually). [Figure 4B] 1 shows the dose-response relationship of calcium responses induced by nicotinamide in cells co-expressing Nanchung and Inactive (Nan+Iav) or Nanchung and Water witch (Nan+Wtrw). [Figure 5A]Figure 1 shows the saturable binding of [H]-afidopyropene to Nanchung (Nan) protein alone, Water witch (Wtrw) protein alone, and Nanchung protein and Water witch protein (Nan + Wtrw). Binding affinity (Kd) is shown in parentheses. [Figure 5B] Scatchard transformation of saturable binding and increased binding affinity of the complex formed by Nanchung protein and Water witch protein (Nan+Wtrw) compared to Nanchung protein (Nan) individually are shown. [Figure 6A] 1 shows the competition between [ 3 H]-afidopyropene and unlabeled pymetrozine for binding to a complex formed by Nanchung protein and Inactive protein, or to a complex formed by Nanchung protein and Water witch protein. [Figure 6B] 1 shows the competition between [3H]-afidopyropene and unlabeled nicotinamide for binding to the complex formed by Nanchung protein and Inactive protein or the complex formed by Nanchung protein and Water witch protein. [Figure 7A] Figures 7A and 7B show activation of Ca2+ mobilization by the plant-derived interferents cinnamaldehyde (A) and allyl isothiocyanate (B) in mammalian cells expressing Water witch protein (Wtrw), cells co-expressing Nanchung and Water witch protein (Nan+Wtrw), and cells co-expressing Inactive and Water witch (Ca2+ mobilization was not activated in parental mammalian cells, cells expressing Nancung (Nan), Inactive (Iav), and cells co-expressing Nanchung and Inactive protein (Nan+Iav)). [Figure 7B] Continued from Figure 7A. [Figure 8]The amino acid sequence between the homologous regions of the Inactive (Iav) and Water witch (Wtrw) proteins is shown, indicating limited (22%) identity. The horizontal bar indicates the position of the predicted transmembrane domain. [Figure 9A] The sequence alignment of Nanchung proteins (SEQ ID NOs: 64-90) is shown in order of appearance, respectively. [Figure 9B] Continued from Figure 9A. [Figure 9C] Continued from Figure 9B. [Figure 9D] Continued from Figure 9C. [Figure 9E] Continued from Figure 9D. [Figure 9F] Continued from Figure 9E. [Figure 9G] Continued from Figure 9F. [Figure 9H] Continued from Figure 9G. [Figure 9I] Continued from Figure 9H. [Figure 9J] Continued from Figure 9I. [Figure 9K] Continued from Figure 9J. [Figure 10A] The sequence alignment of Water witch proteins (SEQ ID NOs: 91 to 126) is shown in order of appearance. [Figure 10B] Continued from Figure 10A. [Figure 10C] Continued from Figure 10B. [Figure 10D] Continued from Figure 10C. [Figure 10E] Continued from Figure 10D. [Figure 10F] Continued from Figure 10E. [Figure 10G] Continued from Figure 10F. [Figure 10H] Continued from Figure 10G. [Figure 10I] Continued from Figure 10H. [Figure 10J] Continued from Figure 10I. [Figure 11] The alignment tree of Nanchung proteins is shown. [Figure 12]The alignment tree of the Water witch protein is shown. DETAILED DESCRIPTION OF THE INVENTION

[0027] Detailed Description of the Drawings and Presently Preferred Embodiments I. General Considerations Transient receptor potential (TRP) channels constitute a large and important class of channels involved in regulating cellular homeostasis (Wu et al., 2010). The present invention provides methods and compositions that identify or contain one or more compounds or endogenous modulators that bind to and / or modulate at least one TRP family member, including, but not limited to, TRPV and TRPA channels in insects.

[0028] Specifically, the present invention provides methods for identifying candidate compounds or endogenous modulators that can directly bind to and regulate the protein complex consisting of the TRPV protein Nanchung and the TRPA protein Water witch.

[0029] The present invention further provides methods for identifying candidate compounds capable of directly modulating the TRPA protein Water witch.

[0030] Modulating insect TRPV and / or TRPA channels can regulate intracellular calcium homeostasis, sodium homeostasis, intracellular calcium level, membrane polarization (resting membrane potential), and / or cation level. Compounds that can regulate one or more insect TRPV and / or insect TRPA proteins, and / or the complex formed by insect TRPV and TRPA proteins, are useful in many aspects, including but not limited to insect control.Knowing whether a compound directly binds to or regulates TRPV proteins and / or TRPA proteins, and / or the complex formed by insect TRPV proteins and TRPA proteins, and knowing whether a compound binds to the same or different sites of insect TRPV proteins and / or insect TRPA proteins, and / or the complex formed by insect TRPV proteins and TRPA proteins, is important for planning insect management and preventing the emergence of resistant insect populations.

[0031] The present invention identifies aphidopyropene, pymetrozine and the endogenous compound nicotinamide that regulate the biological activity of the complex formed by the insect TRPV Nanchung protein and the insect TRPA Water witch protein, with the aim of determining their biological activity by the calcium mobilization assay set out herein.

[0032] The present invention further identifies cinnamaldehyde and allyl isothiocyanate, which modulate the biological activity of TRPA Water witch protein and / or the complex formed by insect TRPV Nanchung protein and insect TRPA Water witch protein, for the purpose of determining the biological activity by the calcium mobilization assay set forth herein.

[0033] The present invention further identifies aphidopyropenes that have affinity for a complex or channel consisting of TRPV Nanchung protein and TRPA Water witch protein, such that one or more of the above compounds may be used as labeled compounds in the methods presented herein for determining the binding affinity of the labeled compounds by the saturation binding assays described herein.

[0034] The present invention further identifies afidopiropene, pymetrozine, and the endogenous modulator nicotinamide as compounds having affinity for a complex or channel consisting of TRPV Nanchung protein and TRPA Water witch, such that one or more of the above compounds may be used as labeled compounds and another compound may be used as an unlabeled compound for purposes of determining the binding affinity of the unlabeled compounds by the competitive binding assays described herein.

[0035] The present invention also provides compositions comprising the candidate compounds identified herein for use as insecticides and / or insect control agents for treating pests, plants, and crops. The present invention further includes classifying the candidate compounds identified herein by their mechanism of action.

[0036] Without being bound by theory, the modulation of insect TRPV protein and / or TRPA protein, and / or the complex formed by insect TRPV protein and insect TRPA protein, induces the signal transduction pathway that leads to the modulation of sensory neurons, and can kill insects, change insect behavior, or repel insects.Therefore, again without being bound by theory, the compound that modulates (for example, activates) insect TRPV protein and / or TRPA protein, and / or the complex formed by insect TRPV protein and insect TRPA protein can be used as insecticide or insect repellent.

[0037] Transient receptor potential (TRP) proteins form cation channels that have been classified into at least seven groups not found in mammals: TRPC (canonical), TRPV (vanilloid), TRPM (melastatin), TRPP (polycystin), TRPML (mucolipin), TRPA (ankyrin), and TRPN (does not possess a mechanosensor potential C ) ( Fowler and Montell, 2013 ).

[0038] All TRP channels exist as tetramers and can function as homotetramers or form heterotetramers with other members of the same or different subfamilies. TRP channels can be regulated by a variety of physical and chemical stimuli and are involved in virtually all sensory modalities (Fowler and Montell, 2013).

[0039] TRPV channels are present in many organisms, including, for example, insects (Drosophila, Tribolium, Pediculus, Culex, and Anopheles), crustaceans (Daphnia pulex), nematodes (Caenorhabditis elegans), mollusks (Aplysia californica, Lottia gigantea), and mammals (humans, mice, rats, monkeys, and chimpanzees).

[0040] Currently, there are six members of the mammalian TRPV family: TRPV1, TRPV2, TRPV3, TRPV4, TRPV5, and TRPV6. Mammalian TRPVs are involved in various sensory functions, including pain perception, temperature, osmotic pressure, touch, and taste (Venkatachalam and Montell, 2007). In addition to physical stimuli, mammalian TRPVs can be activated or inhibited by a variety of endogenous and exogenous compounds (Vriens et al., 2009).

[0041] The C. elegans TRPV family has five members: OCR-1, OCR-2, OCR-3, OCR-4, and OSM-9. C. elegans TRPVs are involved in various sensory functions, including osmosis, olfaction, and mecanosensation (Upadhyay et al., 2016).

[0042] The arthropod, and more specifically insect, TRPV family contains two members, Nanchung (Nanchung) and Inactive (Inactive), which are most closely related to C. elegans OCR-4 and OSM-9, respectively (Matsuura et al., 2009). Nanchung and Inactive may form closely related heteromers. Nanchung and Inactive heteromers are formed exclusively in mechanosensory organs known as chordotonal organs (Nesterov et al., 2015).

[0043] Chordotonal organs are unique to arthropods, located within insect joints, and activated by stretch resulting from the relative rotation of insect body segments (Kavlie and Albert, 2013). Activation of Nanchung and Inactive heteromers by stretch generates neural signals that transduce information about gravity, sound, and the relative position of body segments (Gong et al., 2004, Kim et al., 2003, Nesterov et al., 2015).

[0044] At least three commercially available insecticides, aphidopyropene, pymetrozine, and pyrifluquinazon, overstimulate and ultimately silence Nanchung and Inactive heteromers, disrupting insect coordination, inhibiting feeding, and ultimately leading to death by desiccation and starvation ( Nesterov et al., 2015 ).

[0045] Although afidopyropene, pymetrozine, and pyrifluquinazone can only directly bind to the Nanchung protein, the formation of Nanchung and Inactive heteromers significantly enhances binding affinity (Kandasamy et al., 2017). Because neither Nanchung nor Inactive alone is activated by any of the three insecticides, the formation of Nanchung and Inactive heteromers is essential for biological activity and insecticidal action (Kandasamy et al., 2017; Nesterov et al., 2015).

[0046] Pymetrozine can also function as an insect repellent, although the molecular mechanism of repulsion remains unclear (Chang and Snyder, 2008).

[0047] In addition to insecticides, both insect Nanchung and Inactive, as well as their nematode orthologs, OCR-4 and OSM-9, are activated by nicotinamide, a form of vitamin B3 (Upadhyay et al., 2016). Nicotinamide is absorbed from the diet and actively produced. Similar to the activation of Nanchung and Inactive by insecticides, activation by nicotinamide required coexpression of either insect Nanchung and Inactive or nematode OCR-4 and OSM-9.

[0048] The effect of either insecticides or nicotinamide requires the simultaneous presence of Nanchung and Inactive, but it has not been shown whether the aforementioned insecticides and nicotinamide bind to the same or different sites on the Nanchung and Inactive heteromer.

[0049] In chordotonal organs, Nanchung and Inactive are interdependent; without Inactive, Nanchung is not expressed, and without Nanchung, Inactive is not expressed (Gong et al., 2004; Nesterov et al., 2015). This suggests that the formation of Nanchung and Inactive heteromers is essential for the stability of both Nanchung and Inactive proteins in chordotonal organs. However, in some other insect tissues, Nanchung and Inactive can function independently of each other. Notably, Nanchung, but not Inactive, is required for the detection of food hardness by mechanosensory neurons in the labellum of fruit flies (Jeong et al., 2016). Similarly, Nanchung, but not Inactive, is required for defensive kicking behavior in fruit fly wing margin setae (Li et al., 2016). These observations raise the possibility that Nanchung partners with other TRP channels for protein stability and functional responses in specific tissues.

[0050] The commercially available insecticide flonicamid (Maienfisch, 2012) and its more active derivative, 4-trifluoronicotinamide (TFNA-AM) (Salgado et al., 2014), act on insect chordotonal organs and alter insect feeding behavior, similar to aphidpyropene, pymetrozine, and prifluquinazone. However, these compounds do not directly bind to or modulate insect TRPV channels and are thought to have different, as yet unknown, molecular targets (Kandasamy et al., 2017).

[0051] One TRPA gene has been identified in mammals (Wu et al., 2010), and six TRPA genes have been identified in insects (Matsuura et al., 2009). Various members of the TRPA family are present in various insect species. Fruit flies have four TRPA channels: TRPA1, Painless, Pyrexia, and Water witch.

[0052] Insect TRPA1 channels are involved in the sensation of temperature, mechanical stimuli, and light ( Fowler and Montell, 2013 ).

[0053] Insect TRPA1 channels can be activated by several plant-derived insecticides and / or repellents, including cinnamaldehyde and allyl isothiocyanate ( Kang et al., 2010 ).

[0054] Cinnamaldehyde and allyl isothiocyanate regulate TRPA1 channels by covalently modifying specific cysteine ​​and lysine amino acids in the TRPA1 protein ( Hinman et al., 2006 , Macpherson et al., 2007 ).

[0055] Painless is involved in the sensation of temperature, mechanical stimuli, and gravity (Fowler and Montell, 2013).

[0056] Pyrexia is involved in the sense of temperature and gravity (Fowler and Montell, 2013).

[0057] Water witch is involved in the discrimination between moist and dry air in fruit flies (Liu et al., 2007) and recovery from tonic immobility in beetles (Kim et al., 2015). In fruit flies, water witch is expressed in hydroreceptors (Liu et al., 2007) and chordotonal organs in the second antennal segment (Senthilan et al., 2012). Replacing a conserved glutamate with lysine (E754K) in fruit fly water witch (Cooper et al., 2008), a point mutation that abolishes ion conductance by other TRP channels (Li et al., 2007), impairs sound perception in fruit flies (Senthilan et al., 2012).

[0058] Modulation of the fruit fly TRPA-1 channel is thought to be the primary mechanism of action of the plant-derived pesticides cinnamaldehyde and allyl isothiocyanate ( Kang et al., 2010 ).

[0059] Based on amino acid composition, Painless, Pyrexia, and Water witch were predicted to be unresponsive to cinnamaldehyde and allyl isothiocyanate ( Kang et al., 2010 ).

[0060] Cinnamaldehyde and allyl isothiocyanate were unable to directly modulate Painless (Sokabe et al., 2008).

[0061] So far, no chemical modulators of water witch have been reported.

[0062] The TRPA channels Water with and TRPV kernel Inactive belong to different subfamilies, with Water witch having only 22% identity to Inactive (Fig. 8). However, we discovered that the TRPV protein Nanchung forms a complex with the TRPA protein Water witch that is similar to the previously described complexes of the TRPV protein Nanchung and the TRPV protein Inactive.

[0063] Furthermore, we discovered that the protein complex formed by the TRPV protein Nanchung and the TRPA protein Water witch can be regulated by the commercially available insecticides aphidpiropen and pymetrozine, similar to the complex formed by the TRPV proteins Nanchung and Inactive.

[0064] Furthermore, we discovered that the protein complex formed by the TRPV protein Nanchung and the TRPA protein Water witch can be regulated by the endogenous agonist nicotinamide, similar to the complex formed by the TRPV proteins Nanchung and Inactive.

[0065] Water witch protein is predicted not to interact with cinnamaldehyde and allyl isothiocyanate, which are electrophilic plant-derived insecticides and / or pesticides (Kang et al., 2010). However, we discovered that Water witch can regulate cinnamaldehyde and allyl isothiocyanate.

[0066] Furthermore, we discovered that the formation of a complex between the TRPV protein Nanchung and the TRPA protein Water witch significantly increases the binding affinity of the commercially available insecticide aphidpiropene.

[0067] Furthermore, we discovered that the commercially available insecticides afidopyropene and pymetrozine and the endogenous agonist nicotinamide bind to the same site in a complex containing the TRPV protein Nanchung and the TRPV protein Inactive.

[0068] Furthermore, we discovered that the commercially available insecticides aphidpiropene and pymetrozine and the endogenous agonist nicotinamide bind to the same site in a complex containing the TRPV protein Nanchung and the TRPA protein Water witch.

[0069] Furthermore, the inventors have developed methods to identify additional compounds that directly bind to and modulate a complex containing the TRPV protein Nanchung and the TRPA protein Water witch.

[0070] Furthermore, the present inventors have developed methods to identify additional compounds that directly modulate Water witch proteins or the complex formed by Nanchung proteins and Water witch proteins.

[0071] Identification of such compounds may lead to the identification of suitable compounds that may function as either insecticides or insect repellents and may be useful in both pest management programs, plant protection, crop protection, and public health.

[0072] II. Definition: Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently disclosed subject matter, representative methods, devices, and materials are now described.

[0073] The terms "a," "an," and "the" when used in this application, including the claims, refer to "one or more." Thus, for example, a reference to "a cell" ("a mammalian cell") includes a plurality of such cells (e.g., a plurality of mammalian cells in culture).

[0074] As used herein, the term "about," when referring to a value or amount of mass, weight, time, volume, concentration, or percentage, is meant to encompass variations from the specified amount of, in some embodiments, ±20%, in some embodiments, ±10%, in some embodiments, ±5%, in some embodiments, ±1%, in some embodiments, ±0.5%, and in some embodiments, ±0.1%, where such variations are appropriate for performing the disclosed methods.

[0075] The terms "insect," "insect pest," or "plant insect pest" refer to any one of numerous, usually small, arthropods of the class Insecta that are known to associate with plants and, as a result of that association, have a deleterious effect on the health and vitality of the plant. As used herein, the term plant includes whole plants and plant parts such as roots, stems, leaves, and seeds, as well as cells and tissues within the plant or plant parts. The terms "insect," "insect pest," or "plant insect pest" are used interchangeably throughout this application.

[0076] The terms "pest" and / or "pest species" refer to any one of numerous, usually small, arthropods of the class Insecta that are known to associate with plants and, as a result of that association, have a deleterious effect on the health and vitality of the plants. These terms are further intended to include mollusks, nematodes, and other arthropod classes that are known to associate with plants and, as a result of that association, have a deleterious effect on the health and vitality of the plants. The term further includes arthropods, gastropods, and nematodes, and includes insects from the order Lepidoptera, such as Achroia grisella, Acleris spp. (e.g., A. fimbriana, A. gloverana, A. variana); Acrolepiopsis assectella, Acronicta major, Adoxophyes spp. (e.g., A. cyrtosema, A. orana; Aedia leucomelas), Agrotis spp. (e.g., A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); Alabama argillacea, Aleurodicus dispersus, Alsophila pometaria, Ampelophaga rubiginosa, Amyelois transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp. (e.g. A. gemmatalis); Apamea spp, Aproaerema modicella, Archips spp. (e.g. A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyrethia conjugella, Argyroploce spp., Argyrotaenia spp. (e.g., A.velutiOntop);Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp. Cacoecia spp. (Cacoecia spp. C. muriNanchunga C. podana); Cactoblastis cactorum Cadra cautella Calingo braziliensis Caloptilis theivora Capua reticulana Carposina spp. (including C. niponensis, C. sasakii); Cephus spp., Chaetocnema aridula, Cheimatobia brumata, Chilo spp. (including C. indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp., Colias eurytheme, Conopomorpha spp.、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus); Desmia funeralis, Diaphania spp. (including D. nitidalis, D. hyalinata); Diatraea grandiosella, Diatraea saccharalis. Diphthera festiva subsignaria spp. Eupoecilia ambiguella Euproctis chrysorrhoea Euxoa spp. Evetria bouliana Faronta albilinea Feltia spp spp. (from G. funebrana, G. molesta, G. inopinata);Halysideta spp.American Harrisina, Hedylepta spp., Helicoverpa spp spp. (including H. assulta, H. subflexa, H. virescens); Hellula spp. (including H. undalis, H. rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria、Lambdina fiscellaria lugubrosa、Lamprosema indicata、Annoying Laspeyresia、Leguminivora glycinivorella、Lerodea eufala、Leucinodes orbonalis、Leucoma salicis、Leucoptera spp.(from L. coffeella, L. scitella);Leuminivora lycinivorella, Lithocolletis blancardella, Lithophane antennata, Llattia octo (=Amyna axis), Lobesia botrana, Lophocampa spp., and Loxagrotis albicosta, Loxostege spp. (including L. sticticalis, L. cereralis);monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta); Marasmia spp., Marmara spp., Maruca testulalis, Megalopyge lanata, Melanchra picta, Melanitis leda, Mocis spp. (e.g., M.lapites, M.repanda); Mocis latipes, Monochroa fragariae, Mythhimna separata, Nemapogon cloacella, Neoleucinodes elegantalis, Nepytia spp., Nymphula spp., Oiketicus spp., Omiodes indicata, Omphisa anastomosalis, Operophtera brumata, Orgyia pseudotsugata, Oria spp., Orthaga thyrisalis, Ostrinia spp. (eg O. nubilalis); Oulema oryzae、 Paleacrita vernata、 Panolis flammea、 Parnara spp.、 Papaipema nebris、 Papilio cresphontes、 Paramyelois transitella、 Paranthrene regalis、 Paysandisia archon、 Pectinophora spp. (eg P. gossypiella); Peridroma saucia、 Perileucoptera spp.、 such as P. coffeella; Phalera bucephala、 Phryganidia californica, Phthorimaea spp. (for example, P.operculella);Phyllocnistis citrella, Phyllonorycter spp. (e.g., P.blancardella, P.crataegella, P.issikii, P.ringoniella);Pieris spp. (e.g., P.brassicae, P.rapae, P.napi);Pilocrocis tripunctata, Plathypena scabra, Platynota spp. (e.g., P.flavedana, P.idaeusalis, P.stultana);Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp., Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone. Pseudaletia spp. (including P. sequax and P. unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana, Sabulodes aegrotata, Schizura concinna, and Schoenobius spp. Schreckensteinia festaliella, Scirpophaga spp. (S. incertulas, S. innotata); Scotia segetum, Sesamia spp., S. inferens, Seudyra subflava, Cytotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.ni); Tuta (= Scrobipalpula) absoluta, Udea spp. (e.g. U. rubigalis, U. rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, e.g. Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A. anxius, A. planipennis, A. sinuatus); Agriotes spp. (e.g. A. fuscicollis, A. lineatus, A. obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A. glabripennis); Anthonomus spp. (e.g. A. eugenii, A. grandis, A. pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A. linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g. B. lentis, B. pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C. assimilis, C. napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E.similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida、Megaplatypus mutates、Megascelis spp.、Melanotus communis、Meligethes spp.(eg M.aeneus); Melolontha spp.(eg M.hippocastani、M.melolontha);alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P. helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R.billineatus, R.ferrugineus, R.palmarum, R.phoenicis, R.vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S.granaria, S.oryzae, S.zeamais); Sphenophorus spp. (e.g., S.levis); Stegobium paniceum, Sternechus spp. (e.g., S.subsignatus); Strophomorphus ctenotus, Symphyletes spp.Tanymecus spp. Tenebrio molitor Tenebrioides mauretanicus Tribolium spp spp.(including X.pyrrhoderus);and Zabrus spp.(including Z.tenebrioides);Diptera species include Aedes spp.(including A.aegypti, A.albopictus, and A.vexans);Anastrepha ludens, Anopheles spp.(including A. albimanus, A. crucians, A. freeborni, A. gambiae, A. leucosphyrus, A. maculipennis, A. minimus, A. quadrimaculatus, A. sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. discalis, Chrysops silacea, Cochliomyia spp. (e.g., C. hominivorax); Contarinia spp. (e.g., C. sorghicola); Cordylobia anthropophaga, Culex spp. (e.g., C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g., D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g., D. suzukii), Fannia spp. (e.g., F. canicularis); Gastraphilus spp. (e.g., G. intestinalis); Geomyza tipunctata, Glossina Spp. spp. (for example, L. sativae, L. trifolii); Lucilia spp. (for example, L. caprina, L. cuprina, L. sericata); Lycoria pectoralis, Mansonia titillanus, Mayetiola spp. (for example, M. destructor); Musca spp. (for example, M. autumnalis, M.domestica); Muscina stabulans, Oestrus spp. (e.g., O.ovis); Opomyza florum, Oscinella spp. (e.g., O.frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis、Tipula oleracea、Tipula paludosa、and Wohlfahrtia spp.;、Microcephalothrips abdominalis、Neohydatothrips samayunkur、Pezothrips kellyanus、Rhipiphorothrips cruentatus、Scirtothrips spp. spp. (for example, T.images, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, T.tabaci); spp.(eg、A.rapidus、A.superbus); Aeneolamia spp.、Agonoscena spp.、Aulacorthum solani、Aleurocanthus woglumi、Aleurodes spp., Aleurodicus disperses, Aleurolobus barodensis, Aleurothrixus spp., Amrasca spp., Anasa tristis, Antestiopsis spp., Anuraphis cardui, Aonidiella spp., Aphanostigma piri, Aphidula nasturtii, Aphis spp. (e.g., A.craccivora, A.fabae, A.forbesi, A.gossypii, A.grossulariae, A.maidiradicis, A.pomi, A.sambuci, A.schneideri, A.spiraecola); Arboridia apicalis, Arilus critatus, Aspidiella spp., Aspidiotus spp., Atanus spp.、Aulacaspis yasumatsui、Aulacorthum solani、Bactericera cockerelli(Paratrioza cockerelli)、Bemisia spp.(eg B.argentifolii、B.tabaci(Aleurodes tabaci)); Blissus spp.(eg B.leucopterus); spp.、Brachycorynella asparagi、Brevicoryne brassicae、Cacopsylla spp. fragaefolii、Chionaspis tegalensis、Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila, Cimex spp. (e.g., C. hemipterus, C. lectularius); Coccomytilus halli, Coccus spp. (e.g., C. hesperidum, C. pseudomagnoliarum); Corythucha arcuata, Creontiades dilutus, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasynus piperis, Dialeurodes spp. (e.g., D.citrifolii); Dalbulus maidis、Diaphorina spp. spp. (for example, D.cingulatus, D. intermedius); Dysmicoccus spp., Edessa spp., Geocoris spp., Empoasca spp. (for example, E. fabae, E. solana); Epidiaspis leperii, Eriosoma spp. (for example, E. lanigerum, E. pyricola); spp. (e.g., E.heros, E.impictiventris, E.servus); Fiorinia theae, Geococcus coffeae, Glycaspis brimblecombei, Halyomorpha spp. (e.g., H.halys); Heliopeltis spp., Homalodisca vitripennis (=H.coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp. (e.g., I.purchase); Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp. (e.g., L.ulmi); Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp. (e.g., L.hesperus, L.lineolaris, L.pratensis); Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp. (e.g., M.rosae, M.avenae, M.euphorbiae); Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Mela. naphis sacchari、Melanocallis(=Tinocallis)caryaefoliae、Metcafiella spp.、Metopolophium dirhodum、Monellia costalis、Monelliopsis pecanis、Myzocallis coryli、Murgantia spp.Myzus spp.(from M. ascalonicus, M. cerasi, M. nicotianae, M. persicae, M. varians);Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp. (including N. malayanus, N. nigropictus, N. parvus, and N. virescens); Nezara spp. (including N. viridula); Nilaparvata lugens, Nysius huttoni, Oebalus spp.(O.pugnax);Oncometopia spp.Orthezia praelonga Oxycaraenus hyalinipennis Parabemisia myricae Parlatoria spp.Parthenolecanium spp.(P. corni, P. persicae);Pemphigus spp.(P. bursarius, P. populivenae);Peregrinus maidis, Perkinsiella saccharicida, Phenacoccus spp. (including P. aceris, P. gossypii); Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp. (including P. devastatrix), Piesma quadrata, Piezodorus spp.(including P.guildinii);Pinnaspis aspidistrae;Planococcus spp.(including P.citri);P.ficus); Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudacysta persea, Pseudaulacaspis pentagona, Pseudoococcus spp. (e.g., P. comstocki); Psylla spp. (e.g., P. mali); Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp. (e.g., Q. perniciosus); Quesada gigas, Rastroccus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp. (e.g., R. pseudobrassicas, R. insertum, R. maidis, R. padi); Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulatus, Sitobion avenae, Sogata spp., Sogatella furcifera, Solubea insularis, Spissistilus festinus (=Stictocephala festina); Stephanitis nashi, Stephanitis pyrioides, Stephanitis takeyai, Tenalaphara malayensis, Tetraleurodes perseae, Therioaphis maculate, Thyanta spp. (e.g., T. accerra, T. perditor); Tibraca spp., Tomaspis spp., Toxoptera spp. (e.g., T. aurantii); Trialeurodes spp.(including T. abutilonea, T. ricini, and T. vaporariorum); Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp. (including U. citri, U. yanonensis); and Viteus vitifolii;The list of Hymenoptera includes Acanthomyops interjectus, Athalia rosae, Atta spp (including A. capiguara, A. cephalotes, A. cephalotes, A. laevigata, A. robusta, A. sexdens, A. texana), Bombus spp., Brachymyrmex spp., and Camponotus spp (from C. floridanus, C. pennsylvanicus, C. modoc); Cardiocondyla nuda, Chalibion ​​sp, Crematogaster spp, Dasymutilla occidentalis, Diprion spp, Dolichovespula maculata, Dorymyrmex spp, Dryocosmus kuriphilus , Formica spp , Hoplocampa spp .(including H. minutea , H. testudinea ); invasive, Monomorium spp(from M.pharaonis), Monomorium, Nylandria fulva, Pachycondyla chinensis, Paratrechina longicornis, Paravespula spp(such as P. germanica, P. pennsylvanica, P. vulgaris);Pheidole spp(such as P.megacephala);Pogonomyrmex spp(such as P.barbatus),P.californicus), Polistes rubiginosa, Prenolepis impairs, Pseudomyrmex gracilis, Schelipron spp. spp (for example, T.caespitum, T.bicarinatum), Vespa spp. spp (for example, G.africana、G.gryllotalpa); angulifera, Locusta spp. (e.g., L.migratoria, L.pardalina); Melanoplus spp. (e.g., M.bivittatus, M.femurrubrum, M.mexicanus, M.sanguinipes, M.spretus); Nomadacris septemfasciata, Oedaleus senegalensis, Scapteriscus spp., Schistocerca spp. (e.g., S.americana, S.gregaria), Stemopelmatus spp., Tachycines asynamorus, and Zonozerus variegatus; pests from the class Arachnida, for example mites (Acari) of the families Argasidae, Ixodidae, and Sarcoptidae, for example Amblyomma spp. (A. americanum, A. variegatum, A. maculatum, etc.), Argas spp. (for example, A. persicu), Boophilus spp. (for example, B. annulatus, B. decoloratus, B. microplus), Dermacentor spp. (for example, D. silvarum, D. andersoni, D. variabilis), Hyalomma spp. (for example, H. truncatum), Ixodes spp. (e.g. I. ricinus, I. rubicundus, I. scapularis, I. holocyclus, I. pacificus), Rhipicephalus sanguineus, Ornithodorus spp. (e.g. O. moubata, O. hermsi, O. turicata), Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes Sarcoptes spp. (e.g. S. Scabiei); and Aceria spp (e.g. A. sheldoni, A. anthocoptes), Acallitus spp; Aculops spp. (e.g., A. lycopersici, A. pelekassi); Aculus spp. (e.g., A.schlechtendali); Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; Eriophyidae, including Eriophytes ribis and Eriophyes spp. (e.g. Eriophyes sheldoni); Tarsonemidae, including Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp. and Steneotarsonemus spinki; Tenuipalpidae, including Brevipalpus spp. (e.g. B. phoenicis); Eotetranychus spp., Eutetranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp. (e.g. T. cinnabarinus, T. evansi, T. kanzawai, T, pacificus, T. phaseulus, T. telarius and T. urticae); Bryobia praetiosa; Panonychus spp. (e.g. P. ulmi, P. citri); Metatetranychus spp. The Tetranychidae family includes Oligonychus spp. (e.g., O. pratensis, O. perseae); Vasates lycopersici; Raoiella indica; Carpoglyphidae family includes Carpoglyphus spp.; Penthaleidae spp. (e.g., Halotydeus destructor); Demodicidae family includes Demodex spp.; Trombicidea family includes Trombicula spp.; Macronyssidae family includes Ornothonyssus spp.; Pyemotidae family includes Pyemotes tritici; Tyrophagus putrescentiae; Acaridae family includes Acarus siro; Latrodectus mactans, Tegenaria agrestis, Chiracanthium spp., Lycosa spp., Achaearanea tepidariorum, and Loxosceles reclusa; pests from the phylum Nematoda, for example, plant-parasitic nematodes (e.g., root-knot nematodes Meloidogyne spp. (e.g., M. hapla, M. incognita, M. javanica); cyst nematodes Globodera spp. (e.g., G. rostochiensis); Heterodera spp. (e.g., H. avenae, H. glycines, H. schachtii, H. trifolii); seed gall nematodes Anguina spp.; stem and foliar nematodes Aphelenchoides spp. (e.g., A. besseyi); sting nematodes Belonolaimus spp. (e.g., B. longicaudatus); pine nematodes Bursaphelenchus spp. (e.g., B. lignicolus, B. xylophilus); Ring nematodes Criconema spp.Criconemella spp. (e.g., C. xenoplax and C. ornata); and Criconemoides spp. (e.g., Criconemoides informis; Mesocriconema spp.; Stem and bulb nematodes Ditylenchus spp. (e.g., D. destructor, D. dipsaci); Awl nematodes Dolichodorus spp.; Spiral nematodes Heliocotylenchus multicinctus; Sheath and sheathoid nematodes Hemicycliophora spp. and Hemicriconemoides spp.; Hirshmanniella spp.; Lance nematodes Hoploaimus spp.; False rootknot nematodes Nacobbus spp.; Needle nematodes Longidorus spp. (e.g., L. elongatus); Lesion nematodes Pratylenchus spp. (e.g., P. brachyurus, P. neglectus, P. penetrans, P. curvitatus, P. goodeyi); Burrowing nematodes Radopholus spp. (e.g., R. similis; Rhadopholus spp.; Rhodopholus spp.; Reniform nematodes Rotylenchus spp. (e.g., R. robustus, R. reniformis); Scutellonema spp.; Stubby-root nematodes Trichodorus spp. (e.g., T. obtusus, T. primitivus); Paratrichodorus spp. (e.g., P.minor); Stunt nematodes Tylenchorhynchus spp. (e.g., T. claytoni, T. dubius); Citrus nematodes Tylenchulus spp. (e.g., T. semipenetrans); Dagger nematodes Xiphinema spp.; and other plant-parasitic nematode species; insects from the order Isoptera, for example, Calotermes flavicollis, Coptotermes spp. (e.g., C. formosanus, C. gestroi, C. acinaciformis); Cornitermes cumulans, Cryptotermes spp. (e.g., C. brevis, C. cavifrons); Globitermes sulfureus, Heterotermes spp. (e.g., H. aureus, H. longiceps, H. tenuis); Leucotermes flavipes, Odontotermes spp., Incisitermes spp (e.g. I. minor, I. Snyder); Marginitermes hubbardi, Mastotermes spp (e.g. M. darwiniensis), Neocapritermes spp (e.g. N. opacus, N. parvus); Neotermes spp, Procornitermes spp, Zootermopsis Insects from the order Blattaria, e.g. Blatta spp (e.g. B. orientalis, B. lateralis); Blattella spp (e.g., B. asahinae, B. germanica); Leucophaea maderae, Panchlora nivea, Periplaneta spp (e.g., P.americana, P. australasiae, P. brunnea, P. fuligginosa, P. japonica); Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnoscelus surinamensis; insects from the order Siphonoptera, for example, Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp. (e.g., C. felis, C. canis), Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus; insects from the order Thysanura, for example, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica; pests from the class Chilopoda, for example, Geophilus spp., Scutigera spp. (e.g., Scutigera coleoptrata); pests from the class Diplopoda, for example, Blaniulus guttulatus, Julus spp., Narceus spp.; pests from the class Symphyla, for example, Scutigerella immaculata; insects from the order Dermaptera, for example, Forficula auricularia; insects from the order Collembola, for example, Onychiurus spp. (e.g., Onychiurus armatus); pests from the order Isopoda, for example, Armadillidium vulgare, Oniscus asellus, Porcellio scaber; insects from the order Phthiraptera, for example, Damalinia spp., Pediculus spp. (e.g., Pediculus humanus capitis, Pediculus humanus corporis, Pediculus humanus humanus); Pthirus pubis, Haematopinus spp.(e.g., Haematopinus eurysternus, Haematopinus suis); Linognathus spp. (e.g., Linognathus vituli); Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes spp. Examples of further pest species that may be controlled by the compounds and / or compositions identified herein include the phylum Mollusca, the class Bivalvia, e.g., Dreissena spp.; the class Gastropoda, e.g., Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea canaliclata, Succinea spp.; the class Helminthi, e.g., Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia Heterakis spp., Hymenolepis Nanchunga, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca volvulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborni, Strongyloides stercora lis, Stronyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella b. ritovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichuria, and Wuchereria bancrofti.

[0077] The terms "non-pest" and / or "non-target" species refer to all other animals that are not classified as "pests" and / or "pest species" as defined above.

[0078] The term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single- or double-stranded form, composed of monomers (nucleotides) containing a sugar, a phosphate, and a base that is either a purine or a pyrimidine. Unless otherwise limited, the term encompasses known analogues of natural nucleotides, conservatively modified variants thereof, complementary sequences, and nucleic acids containing degenerate codon substitutions that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides.

[0079] The terms "nucleic acid" or "nucleic acid molecule" can also be used interchangeably with gene, nucleic acid molecule, nucleic acid fragment, nucleic acid segment, or gene encoded by a polynucleotide, open reading frame (ORF), cDNA, and mRNA.

[0080] The term "gene" is broadly used to refer to any segment of nucleic acid associated with a biological function. Thus, genes include coding sequences and / or regulatory sequences required for their expression. For example, "gene" refers to a nucleic acid fragment that expresses mRNA, functional RNA, or a specific protein, including regulatory sequences. "Gene" also includes unexpressed DNA segments that, for example, form recognition sequences for other proteins. "Gene" can be obtained from a variety of sources, including cloning from a source of interest or synthesis from known or predicted sequence information, and can include sequences designed to have desired parameters.

[0081] The terms "gene delivery" or "gene transfer" refer to methods or systems for reliably introducing foreign DNA into target cells, including injection, transduction, transfection, transformation, and electroporation. Such methods can result in transient or long-term expression of the gene.

[0082] The term "transduction" refers to the delivery of a DNA molecule into a recipient cell either in vivo or in vitro, for example, via a replication-defective viral vector such as an adenoviral vector.

[0083] The term "transfection" refers to the uptake of foreign DNA by mammalian cells. A cell is "transfected" when exogenous DNA is introduced across the cell plasma membrane. Transfection can be used to introduce one or more exogenous DNA moieties, such as plasmid vectors and other nucleic acid molecules, into suitable cells. The term refers to both stable and transient uptake of genetic material.

[0084] The terms "transformation" and / or "transfection" refer to a process for introducing heterologous DNA into a cell. A transformed cell is understood to encompass not only the end product of the transformation / transfection process but also its transgenic progeny.

[0085] The term "injection" refers to the process for introducing heterologous DNA or RNA into cells using a syringe and needle.

[0086] The term "vector" refers to any genetic element, such as a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc., that is capable of replication and transfer of genetic sequences between cells when associated with appropriate control elements, such as a helper virus. Thus, the term includes cloning and expression vehicles, as well as replication-defective viral vectors.

[0087] The terms "expression vector" and "expression cassette" refer to nucleic acid molecules capable of directing the expression of a particular nucleotide sequence in an appropriate host cell, typically comprising a promoter operably linked to a nucleotide sequence of interest operably linked to a termination signal. It also typically includes sequences required for proper translation of the nucleotide sequence. The coding region usually encodes a polypeptide of interest, but can also encode a functional RNA of interest (e.g., antisense RNA or non-translated RNA) in either the sense or antisense orientation. An expression cassette containing a nucleotide sequence of interest may be chimeric, meaning that at least one of its components is heterologous to at least one of its other components. An expression cassette may also be naturally occurring, but may also be derived in a recombination form useful for heterologous expression. Typically, however, the expression cassette is heterologous to the host. That is, the particular DNA sequence of the expression cassette does not naturally occur in the host cell, but was introduced into the host cell or an ancestor of the host cell by a transformation event. Expression of the nucleotide sequence in the expression cassette may be under the control of a constitutive promoter or an inducible promoter that initiates transcription only when the host cell is exposed to a particular external stimulus. In the case of multicellular organisms such as plants, promoters can be specific to particular tissues, organs, or developmental stages.

[0088] The term "adenovirus" refers to vectors derived from adenovirus serotypes. Adenoviruses are double-stranded DNA viruses capable of infecting a wide range of mammalian cells, both dividing and non-dividing. Adenoviral DNA is linear and approximately 36,000 bp long. The most widely used adenoviral vectors for gene delivery are replication-deficient vectors in which the E1 and E3 regions of the adenoviral genome have been deleted. E1-deleted viruses grow in complementing cells such as HEK-293, which provide E1-encoded proteins in trans. E1 / E3-deleted adenoviruses can accommodate up to 7.5 kb of foreign DNA. The classical method for integrating foreign DNA into the adenoviral genome involves homologous recombination in mammalian cells. As described in this example, a more efficient method involves homologous recombination in Escherichia coli between a large plasmid containing most of the adenoviral genome and a small shuttle plasmid. The shuttle plasmid contains an expression cassette flanked by sequences homologous to the target region within the viral genome. The recombinant Ad genome is then linearized by restriction digestion and transfected into mammalian cells complemented with E1 to produce viral particles (Douglas JT., Methods Mol Biol. 2004;V246:3-14). To produce adenoviruses containing toxic genes, specialized adenoviral vectors have been developed in which the expression of the foreign gene is suppressed during viral production. One of these systems, the SpeAd Ad-teto system, is offered, for example, by Welgen, Inc.

[0089] The term "tetracycline-controlled transactivator" (tTA) refers to a fusion protein used to control nucleic acid expression in the presence or absence of doxycycline, tetracycline, and related compounds. tTA includes the Tet repressor (TetR) fused to any domain capable of activating transcription. tTA can include TetR fused to the C-terminal portion of an adenovirus.

[0090] The term "operably linked" refers to two nucleic acid sequences that are physically or functionally related. For example, a promoter or regulatory DNA sequence is said to be "operably linked" to a DNA sequence encoding an RNA or polypeptide when the two sequences are positioned so that the regulatory DNA sequence affects the expression level of the coding or structural DNA sequence. A promoter is also said to be operably linked to a nucleotide sequence if the nucleotide sequence is transcribed when RNA polymerase binds to the promoter under conditions sufficient for transcription.

[0091] The term "isolated," when applied to a nucleic acid or polypeptide, means that the nucleic acid or polypeptide is essentially free from other cellular components with which it is naturally associated. It can be in either a dry or aqueous solution, but can be in a homogeneous state. Homogeneity and molecular separation can be determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high-performance liquid chromatography. A polypeptide that is predominantly a Nanchung species present in a preparation is substantially isolated. The term "isolated" means that the nucleic acid or polypeptide gives rise to essentially one band in an electrophoretic gel. Specifically, it means that the nucleic acid or polypeptide is at least about 50% pure in some embodiments, at least about 85% pure in some embodiments, and at least about 99% pure in some embodiments.

[0092] The terms "label" and "labeled" refer to the attachment of a moiety to a molecule that can be detected by spectroscopic, radiological, or other methods. Thus, the terms "label" or "labeled" refer to the incorporation or attachment (optionally covalently or noncovalently) of a detectable marker into a molecule, such as a biomolecule. Various methods of labeling biomolecules are known in the art and can be used. Examples of biomolecule labels include, but are not limited to, radioisotopes, fluorescent labels, heavy atoms, enzyme labels or reporter genes, chemiluminescent groups, and biotin groups. In some embodiments, labels are attached by spacer arms of various lengths to reduce potential steric hindrance.Available fluorescent probes include fluorescein isothiocyanate; fluorescein dichlorotriazine and fluorinated analogs of fluorescein; naphthofluorescein carboxylic acid and its succinimidyl ester; carboxyrhodamine 6G; pyridyloxazole derivatives; Cy2, 3, 3.5, 5, 5.5, and 7; phycoerythrin; phycoerythrin-Cy conjugates; fluorescent species of succinimidyl esters, carboxylic acids, isothiocyanates, sulfonyl chlorides, and dansyl chlorides, including propionic acid succinimidyl ester and pentanoic acid succinimidyl ester; succinimidyl ester of carboxytetramethylrhodamine; rhodamine Red-X succinimidyl ester; Texas Red sulfonyl chloride; Texas Red-X succinimidyl ester; Texas Red-X sodium tetrafluorophenol ester; Red-X; Texas Red Red dyes; tetramethylrhodamine; Lissamine rhodamine B; tetramethylrhodamine; tetramethylrhodamine isothiocyanate; naphthofluorescein; coumarin derivatives (e.g., hydroxycoumarin, aminocoumarin, and methoxycoumarin); pyrene; pyridyloxazole derivatives; dapoxyl dyes; Cascade Blue and Yellow dyes; benzofuran isothiocyanate; sodium tetrafluorophenol; 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; Alexa fluor (e.g., 350, 430, 488, 532, 546, 555, 568, 594, 633, 647, 660, 680, 700, and 750); isotopes of hydrogen (e.g., tritium isotope, 3H), isotopes of carbon (e.g., 14C); green fluorescent protein; yellow fluorescent protein, or red fluorescent protein. The peak excitation and emission wavelengths vary among these compounds, and the selection of a particular fluorescent probe for a particular application can be based in part on the excitation and / or emission wavelength.

[0093] As used herein, the terms "candidate compound" or "test compound" refer to a group of compounds being screened for their ability to bind to Nanchung protein, Water witch protein, Nanchung and Inactive protein complex, Nanchung and Water witch protein complex, and / or modulate insect TRPV channels and / or insect TRPA channels. Candidate compounds can encompass numerous classes of chemical molecules, such as organic or inorganic small molecules, polysaccharides, biological macromolecules (e.g., peptides, proteins, peptide analogs and derivatives), peptidomimetics, nucleic acids, nucleic acid analogs and derivatives, biological materials such as bacterial, plant, fungal, or animal cells or tissues, naturally occurring or synthetic compositions, or combinations thereof. Generally, candidate compounds can have a molecular weight of approximately 50 to 500,000.

[0094] As used herein, the term "small molecule" refers to a compound that is "natural product-like," although the term "small molecule" is not limited to "natural product-like" compounds. Rather, small molecules are typically characterized by containing several carbon-carbon bonds and having a molecular weight greater than about 50 and less than about 5,000 daltons (5 kD). Preferably, small molecules have a molecular weight of less than 3 kD, even more preferably less than 2 kD, and most preferably less than 1 kD. In some cases, it is preferred that the molecular mass of a small molecule be 700 daltons or less.

[0095] In some embodiments, the candidate compound may be a synthetic molecule. The term "synthetic molecule" refers to a molecule that does not exist in nature.

[0096] In certain embodiments, the candidate compound may be a naturally occurring molecule. Such naturally occurring molecules may be used in purified or unpurified form, i.e., obtained from a biological source. The term "naturally occurring" refers to an entity (e.g., a cell, a biological molecule, etc.) that is found in nature, as distinct from one artificially produced by humans. For example, a polypeptide or nucleotide sequence that exists in an organism in its natural state, without being intentionally modified or isolated by humans in a laboratory, is naturally occurring. Thus, a polypeptide or nucleotide sequence is considered "non-naturally occurring" if it is encoded by or present in a recombiNanchungt molecule, even if the amino acid or nucleic acid sequence is identical to an amino acid or nucleic acid sequence found in nature.

[0097] Depending on the particular embodiment being performed, the candidate compound may be provided free in solution or attached to a carrier or solid support (e.g., beads). A number of suitable solid supports may be used for immobilizing the candidate compound. Examples of suitable solid supports include agarose, cellulose, dextran (i.e., commercially available as Sephadex or Sepharose), carboxymethylcellulose, polystyrene, polyethylene glycol (PEG), filter paper, nitrocellulose, ion exchange resins, plastic film, polyamine methyl vinyl ether maleic acid copolymer, glass beads, amino acid copolymers, ethylene-maleic acid copolymer, nylon, silk, and the like. Furthermore, for the methods described herein, candidate compounds can be screened individually or in groups. Group screening is particularly useful when multiple positive results are not expected for a given group because the hit rate for effective candidate compounds is expected to be low.

[0098] There are millions of possible candidate compounds. Methods for developing small molecule, polymer, and genome-based libraries are known and are described, for example, in Ding, et al. J. Am. Chem. Soc. 124:1594-1596 (2002) and Lynn, et al., J. Am. Chem. Soc. 123:8155-8156 (2001). Many small molecule libraries are known in the art and are commercially available. Commercially available compound libraries are available from ArQule, Pharmacopia, graffinity, Panvera, Vitas-M Lab, Biomol International, Oxford, and others. These libraries can be screened using the screening methods described herein. Chemical compound libraries, such as the 10,000 compound and 86,000 compound chemical compound libraries from the NIH Roadmap, Molecular Libraries Screening Centers Network (MLSCN), can also be used. A comprehensive list of compound libraries can be found at www.broad.harvard.edu / chembio / platform / screening / compound_libraries / index.htm. A chemical library or compound library is a stored collection of chemicals, typically for ultimate use in high-throughput screening or industrial production. A chemical library can simply consist of a set of stored chemicals. Each chemical has associated information stored in some kind of database, such as the chemical structure, purity, quantity, and physicochemical characteristics of the compound.

[0099] The term "modulate" refers to an increase, decrease, or other change in any or all of the chemical and / or biological activities and / or properties of a biomolecule, such as a TRPV or TRPA (e.g., insect TRPV or TRPA) channel of the present invention. Thus, as used herein, the term "modulate" refers to both upregulation (i.e., activation or stimulation) and downregulation (i.e., inhibition or suppression) of such activities or properties. As will be understood by those skilled in the art, modulation of the chemical and / or biological activities and / or properties of a biomolecule, such as a TRPV or TRPA channel, can result from an increase or decrease in the expression of the biomolecule within a cell. Thus, the term "modulate" and its grammatical variations are intended to encompass both direct modulation (e.g., inhibition of the chemical and / or biological activity and / or properties of a polypeptide via binding of an inhibitor to the polypeptide) and indirect modulation (e.g., upregulation or downregulation of the expression of a protein, such as a TRPV or TRPA channel, or inhibition or stimulation of a biomolecule that acts together with the biomolecule of the presently disclosed subject matter to produce a biological effect).

[0100] The term "endogenous modulator" refers to a modulator that occurs or is produced within an organism, tissue, or cell.

[0101] The term "exogenous modulator" refers to a modulator that originates or is produced outside an organism, tissue, or cell.

[0102] The terms "polypeptide," "protein," and "peptide," used interchangeably herein, refer to polymers of the 20 protein amino acids or amino acid analogs, regardless of their size or function. Although "protein" is often used in reference to relatively large polypeptides and "peptide" is often used in reference to small polypeptides, the use of these terms in the art overlaps and varies. As used herein, the term "polypeptide" refers to peptides, polypeptides, and proteins, unless otherwise specified. The terms "protein," "polypeptide," and "peptide" are used interchangeably herein when referring to gene products. Thus, exemplary polypeptides include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments, and other equivalents, variants, and analogs of the foregoing.

[0103] The term "TRP channel" refers to a channel and / or protein complex formed from one or more of Nanchung protein, Water witch protein, and / or Inactive protein. It should be understood that a TRPV channel and / or TRPV protein complex is a type of TRP channel and / or TRP protein complex. Nanchung protein and Inactive protein are types of TRPV protein, which are types of TRP protein. Water witch protein is a type of TRPA protein, which is a type of TRP protein. TRP proteins can be found in insect cell membranes, mammalian cell membranes, derived from cell-free systems, produced by cell culture, and / or produced by cells including, but not limited to, frog eggs, insect cells, and / or mammalian cells genetically modified (containing either or both DNA and / or RNA) to contain genes encoding one or more of Nanchung protein, Water witch protein, and / or Inactive protein. It should be understood that the TRP proteins disclosed herein can be mammalian TRP proteins, arthropod TRP proteins, nematode TRP proteins, and / or molluscan TRP proteins. It should further be understood that Nanchung, Inactive, and Water witch proteins can be found in arthropods, including but not limited to insects, as well as mollusks and / or nematodes. It should be understood that TRP and / or TRPV and / or TRPA channels can be formed from one or more TRPA and / or TRPV protein complexes, from two or more TRPA and / or TRPV protein complexes, from three or more TRPA and / or TRPV protein complexes, and / or from four or more TRPA and / or TRPV protein complexes.

[0104] The term "binding affinity" refers to the ability of a compound to form a bond with a receptor. For purposes of the present invention, equilibrium constants are used to determine the binding affinity of one or more compounds with one or more Nanchung proteins, inactive proteins, Water witch proteins, TRP channels, and / or TRPV channels and / or TRPA channels. The dissociation constant (Kd) is a specific type of equilibrium constant measured in a saturation binding assay of a labeled compound. It is the molar concentration of a labeled compound that binds to half of the receptor sites at equilibrium. The lower the dissociation constant, the higher the binding affinity of the candidate compound to Nanchung proteins, inactive proteins, Water witch proteins, and / or the complex formed by Nanchung and inactive proteins, and / or the complex formed by Nancung and Water witch proteins. The inhibition constant (Ki) is a specific type of equilibrium constant measured using competitive binding between an unlabeled compound and a labeled compound. In the absence of a labeled compound, it is the molar concentration of the competing unlabeled compound that occupies 50% of the receptors at equilibrium. It can be calculated using the Cheng and Prusoff equation: K i =IC 50 / (1+[L] / K d ) (where IC 50 (where [L] is the molar concentration of competing unlabeled compound that reduces specific binding of the labeled compound by 50%). In this equation, [L] is the molar concentration of labeled compound in the assay, and Kd is the equilibrium dissociation constant of the labeled compound. In addition to saturation binding assays, equilibrium dissociation constants, Kd can be measured in several ways, including, but not limited to: 1. Use of association and dissociation rate constants, where Kd = Koff / Kon 2. Use of Fluorescence Polarization Assay 3. Use of Surface Plasmon Resonance 4. Use of Thermophoresis 5. Use of Isothermal Calorimetry 6. Use of Schild analysis

[0105] Surface plasmon resonance and isothermal calorimetry reveal that K d No labeled compound is required to measure K. Furthermore, in the case of competitive assays, d and / or K i (for competitive assays) may be calculated using one or more of the Cheng and Prusoff equation, fluorescence polarization, and / or Schild analysis. The calculated K for a compound d value and K i It is understood that the values ​​should be similar, but due to experimental error the difference in values ​​may be less than 2-fold between two values, or less than 3-fold between two values, depending on the degree of experimental error.

[0106] The terms "high affinity" and / or "high binding affinity" refer to the equilibrium dissociation constant (K d ) and / or the equilibrium inhibition constant (K i ) but 3×10 -7 M or less, 3×10 -8 M or less, and / or 30 x 10 -12 It refers to compounds calculated to be less than or equal to M. d or K i The lower the binding affinity for the compound.

[0107] The term "mode of action" refers to the classification by the Insecticide Resistance Task Force (IRAC) of compounds and / or compositions into appropriate groups based on the compounds and / or compositions (a) exhibiting the same function as other insecticides in that group, and (b) binding directly to the same site on a specific protein, protein complex, and / or protein channel as other compounds and / or compositions in that group.

[0108] The term "insecticide" refers to a substance or agent used to kill insects and other arthropods.

[0109] The term "repellent" refers to a substance or agent used to repel insects and other arthropods.

[0110] III. Screening Methods The present inventors have discovered that insect TRPA Water witch proteins and insect TRPV Nanchung proteins physically associate with each other to form a complex consisting of one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins.

[0111] Furthermore, the present inventors have discovered that a complex consisting of one or more TRPA Water witch proteins and one or more TRPV Nanchung proteins can form an ion channel. Thus, in one embodiment, the present invention relates to a method for determining whether a candidate compound is a modulator of an insect TRPA Water witch channel and / or a complex consisting of a TRPA Water witch channel and a TRPV Nanchung channel.

[0112] Furthermore, the inventors have discovered that ion channels formed by TRPA Water witch proteins, and / or ion channels formed by one or more TRPA Water witch proteins and one or more TRPV Nanchung proteins, may be useful as insecticides and / or repellents, as they may be used to identify compounds that kill insects and / or interfere with insect behavior and / or repel insects.

[0113] Furthermore, the present inventors have discovered that determining whether a candidate compound is a modulator of the insect TRPA Water witch channel and / or the complex consisting of the TRPA Water witch channel and the TRPV Nanchung channel may be useful in classifying the compound's mechanism of action.

[0114] Furthermore, the inventors have determined that the compound aphidopiropene binds with high affinity to a protein complex consisting of one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins.

[0115] The present inventors have further discovered a method for identifying compounds that bind directly to a complex formed by one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins.

[0116] Furthermore, the inventors have found that methods for identifying compounds that directly bind to the complex formed by one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins can be used to identify compounds that kill insects and / or interfere with insect behavior and / or repel insects, and are therefore useful as insecticides and / or repellents.

[0117] Furthermore, the inventors have discovered that determining whether different compounds bind to the same or different sites on the complex consisting of the TRPA Water witch protein and the TRPV Nanchung protein may help classify the compound's mechanism of action.

[0118] The methods identified herein include providing one or more Nanchung proteins and / or one or more Water witch proteins to contact a first cell with a candidate compound, and assaying for modulation of an ion channel formed by a TRPA Water witch protein and / or an ion channel comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins. Modulation of an ion channel formed by a TRPA Water witch protein and / or an ion channel comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins may be assayed using conventional in vitro and in vivo methods well known to those skilled in the art. For example, in certain embodiments, the assaying step may include (1) detecting calcium ion mobilization in the first cell in response to the candidate compound, (2) detecting a membrane potential in the first cell in response to the candidate compound, and (3) comparing calcium ion mobilization in the first cell in the absence of the candidate compound with calcium ion mobilization in the first cell in the presence of the candidate compound. For example, the calcium response may be measured by measuring Ca 2+or by using fluorescent dyes such as Fluo-4 or Fura-2, or fluorescent proteins such as Cameleon or GCaMP6m; (4) by comparing the membrane potential in the first cell in the absence of the candidate compound with the membrane potential in the first cell in the presence of the candidate compound; (5) by comparing calcium ion mobilization in the first cell in the presence of the candidate compound with a calcium ion mobilization reference level indicative of no modulation of ion channels formed by TRPA Water witch proteins and / or ion channels comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins; and / or (6) by comparing the membrane potential in the first cell in the presence of the candidate compound with a membrane potential reference level indicative of no modulation of ion channels formed by TRPA Water witch proteins and / or ion channels comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins; (7) by comparing ionic currents in the first cell in the absence of the candidate compound with ionic currents in the first cell in the presence of the candidate compound using standard electrophysiological methods such as voltage clamp or patch clamp. It should be understood that this same assay can be performed on insect TRPV channels and / or TRPA channels from vertebrates, arthropods, mollusks, nematodes, or any other organism. Such testing allows for determining whether a candidate compound functioned as a modulator in those systems, and thus helps determine the potential environmental impact of using the candidate compound as an insecticide.

[0119] Preferably, the candidate compound is capable of modulating calcium ion mobilization or membrane potential or ionic current in the first cell by at least 10% relative to a reference level, more preferably at least 20% relative to a reference level, more preferably at least 30% relative to a reference level, more preferably at least 40% relative to a reference level, more preferably at least 50% relative to a reference level, more preferably at least 60% relative to a reference level, more preferably at least 70% relative to a reference level, more preferably at least 75% relative to a reference level, more preferably at least 80% relative to a reference level, more preferably at least 85% relative to a reference level, more preferably at least 90% relative to a reference level, more preferably at least 95% relative to a reference level.

[0120] Furthermore, modulation of ion channels formed by TRPA Water witch proteins and / or ion channels comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins can be assayed using a variety of other conventional assays that measure changes in ion flux, including, but not limited to, patch clamp techniques, measurements of whole-cell currents, radiolabeled ion flux assays, and fluorescent assays using membrane potential-sensitive dyes (Zheng et al., 2004).

[0121] Modulation of ion channels formed by TRPA Water witch proteins and / or ion channels comprising one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins may also be assessed using a variety of other in vitro and in vivo assays to determine functional, chemical, and physical effects, for example, to measure binding of insect TRPV and / or TRPA channels to other molecules, including peptides, small organic molecules, and lipids, or to measure insect TRPV and / or TRPA protein and / or mRNA levels, or to measure other aspects of insect TRPV and / or TRPA polypeptides (e.g., transcription levels or physiological changes).

[0122] The inventors have further determined that it would be beneficial to identify compounds that bind directly to the complex formed by one or more TRPV Nanchung proteins and one or more TRPA Water witch proteins, and would therefore be useful as insecticides or pesticides.

[0123] There are two variations of binding assays: saturation assays and competitive assays. Saturation assays can include at least one of the following steps: (1) contacting one or more complexes formed by Nanchung and Water witch proteins with a labeled compound; (2) measuring the amount of labeled compound bound to one or more complexes formed by Nanchung and Water witch proteins; and (3) measuring the equilibrium inhibitor dissociation constant (K) of the labeled compound. d (4) calculating K d (a) Maximum connection (B max and (b) calculating the concentration of the candidate compound required to reach 50% of maximum binding.

[0124] A competitive assay may include one or more of the following steps: (1) contacting one or more complexes formed by the linked Nanchung and Water witch proteins with two compounds, one compound being labeled and the other unlabeled; (2) measuring the amount of labeled compound bound to one or more complexes formed by the Nanchung and Water witch proteins at a single labeled compound concentration and at various unlabeled compound concentrations; (3) determining the concentration (%) of unlabeled compound that produces 50% of the maximal binding of the labeled compound. IC50 (4) determining the equilibrium inhibition constant (K i (4) calculating the Ki, wherein Ki can be determined from (a) the IC50 of the unlabeled compound, (b) the Kd of the labeled compound, and (c) the concentration of the labeled compound.

[0125] Competitive binding assays also determine whether labeled and unlabeled compounds bind to the same or different parts of the complex formed by the Nanchung and Water witch proteins.

[0126] Binding to complexes formed by Nanchung and Water witch proteins can also be determined using one or more of the following methods: (I) thermophoresis, (II) fluorescence polarization assay, (III) surface plasmon resonance, and (IV) isothermal calorimetry.

[0127] Identifying the molecular targets of insecticides is essential for assessing their spectrum, health, and environmental impacts, as well as for insect resistance management. Pymetrozine (PM) and aphidopyropen (AP) are two commercially available insecticides that act by disrupting the coordination and feeding of plant-sucking insects, such as aphids and whiteflies, leading to their death by starvation and desiccation (Casida and Durkin, 2013; Maienfisch, 2012).

[0128] We previously demonstrated that PM and AP act by directly binding to the protein Nanchung, a member of the TRPV ion channel family (Kandasamy et al., 2017). However, binding to Nanchung alone was not sufficient to regulate the ion channel. The biological activity of PM and AP required the simultaneous presence of another TRPV protein, Inactive (Kandasamy et al., 2017; Nesterov et al., 2015). Nanchung and Inactive together formed a functional complex that responded to PM and AP by calcium mobilization and plasma membrane depolarization. Furthermore, while PM and AP could bind to Nanchung in the absence of Inactive, the presence of Inactive significantly increased the binding affinity (Kandasamy et al., 2017).

[0129] The present invention demonstrates that Water Witch can form a functional complex with the TRPV channel Nanchung, similar to the inactive, ankyrin-type (TRPA) channel. The present inventors also found that the complex formed by Nanchung and Water Witch proteins can be activated by PM and AP, similar to the complex formed by Nanchung and Inactive proteins. The inventors also found that the complexes formed by both Nanchung and Inactive proteins and Nanchung and Water Witch proteins can be activated by the endogenous modulator nicotinamide. The inventors also found that co-expression of Nanchung and Water Witch, like co-expression of Nanchung and Inactive proteins, significantly enhanced the binding of aphidpiropen to Nanchung. Thus, the inventors found that both the functional and binding complexes formed by Nanchung and Water Witch proteins are functionally similar to the complex formed by Nanchung and Inactive proteins. This finding was unexpected, as Inactive and Water Witch belong to different types of TRP channels: vanilloid and ankyrin, respectively. Furthermore, based on amino acid sequence alignment, Water witch shares only 22% sequence identity with Inactive (Figure 8). We also found that the endogenous modulator nicotinamide and the exogenous modulators AP and PM have overlapping binding sites. We also found that the channels formed by Water witch protein and those formed by Nanchung and Water witch proteins can be activated by the plant-derived compounds cinnamaldehyde and allyl isothiocyanate, which can function as insecticides and / or repellents. This finding was unexpected, as the primary target of these compounds is thought to be the insect TRPA1 protein.Furthermore, based on the amino acid sequence, Water Witch protein was predicted not to be the target of cinnamaldehyde and isothiocyanate allyl (Kang et al., 2010). Therefore, the present invention identifies methods for determining whether a candidate compound modulates the activity of Water Witch protein and / or the complex formed by Nanchung protein and Water Witch protein. Identification of such modulators allows for the identification of new insecticides and / or insect control agents. Furthermore, the present invention identifies methods for detecting binding, calculating binding affinity, and determining whether a candidate compound directly binds to the same or different sites on the complex formed by Nanchung and Water Witch proteins. Determining the binding location and affinity of candidate compounds allows for the identification of new insecticides and / or insect control agents, facilitates classification of the compound's mechanism of action, and improves the management and control of insects, which may reduce the risk of developing resistant insect populations.

[0130] As mentioned above, candidate compounds can be small organic molecules, small inorganic molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials such as bacteria, plants, fungi, animal cells, animal tissues, and any combination thereof.

[0131] Generally, candidate compounds can be tested at any concentration that can modulate the activity of an ion channel formed by a Water Witch protein and / or a complex comprising one or more Nanchung proteins and one or more Water Witch proteins, and / or demonstrate binding to a complex comprising one or more Nanchung proteins and one or more Water Witch proteins. In some embodiments, candidate compounds can be tested at concentrations ranging from about 0.01 nM to about 1000 mM. In certain other embodiments, compounds can be tested at concentrations ranging from about 100 μM to about 1000 μM. In certain further embodiments, candidate compounds may be tested at 0.05 mM, 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 1.1 mM, 1.2 mM, 1.3 mM, 1.4 mM, 1.5 mM, 1.6 mM, 1.7 mM, 1.8 mM, 1.0 mM, or greater than 2 mM. Other ranges and concentrations of candidate compounds may also be suitable.

[0132] In some embodiments, the candidate compound may be tested at two or more different concentrations. Preferably, the highest concentration tested is at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, at least 75-fold, at least 100-fold, at least 200-fold, or at least 250-fold higher than the lowest concentration used. For example, the candidate compound may be tested at 0.1 mM, 0.5 mM, and 1 mM, with the highest concentration being at least 10 times higher than the lowest concentration used.

[0133] Typically, a complex formed by one or more Water Witch proteins and / or a Nanchung protein and a Water Witch protein can be contacted with a candidate compound for any suitable time before measuring the binding affinity and / or activity of the Water Witch protein and / or a complex containing one or more Nanchung proteins and one or more Water Witch proteins. For example, a complex formed by a Water Witch protein and / or a Nanchung protein and a Water Witch protein can be contacted with a candidate compound for at least 5 seconds, at least 10 seconds, at least 15 seconds, at least 30 seconds, at least 45 seconds, at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 30 minutes, at least 60 minutes, at least 2 hours, at least 3 hours, or more before measuring the activity of the Water Witch protein and / or a complex formed by a Nanchung protein and a Water Witch protein. In some embodiments, activity can be measured at the moment the Water Witch protein and / or a complex formed by a Nanchung protein and a Water Witch protein is contacted with the candidate compound.

[0134] In some embodiments, the binding affinity and / or activity of a Water Witch protein and / or a complex formed by a Nanchung protein and a Water Witch protein may be measured over a period of time. For example, activity may be measured for a period of at least 5 seconds, at least 10 seconds, at least 15 seconds, at least 30 seconds, at least 45 seconds, at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 30 minutes, at least 60 minutes, at least 2 hours, at least 3 hours, at least 4 hours, or more. The measurement period can begin before the Water Witch protein and / or a complex formed by a Nanchung protein and a Water Witch protein comes into contact with a candidate compound, at the moment the Water Witch protein and / or a complex formed by a Nanchung protein and a Water Witch protein first comes into contact with a candidate compound, or can begin a certain period of time after one or more Water Witch proteins and / or a complex formed by a Nanchung protein and a Water Witch protein first comes into contact with a candidate compound. The Water witch protein and / or the complex formed by the Nanchung protein and the Water witch protein may be continuously contacted with the candidate compound while activity and / or binding is being measured.

[0135] In some embodiments, the candidate compound has an effective concentration EC50 of 500 nM or less, 250 nM or less, 100 nM or less, 50 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less for activating and / or binding to Water witch protein and / or the complex formed by Nanchung protein and Water witch protein.

[0136] In some embodiments, the candidate compound has an IC50 of 500 nM or less, 250 nM or less, 100 nM or less, 50 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM or less, or 0.001 nM or less for inhibiting and / or binding to Water witch protein and / or the complex formed by Nanchung protein and Water witch protein.

[0137] In certain embodiments, the candidate compound is a modulator that inhibits the activity of Water witch protein and / or the complex formed by Nanchung protein and Water witch protein.

[0138] In certain other embodiments, the candidate compound is a modulator that activates Water witch protein and / or the complex formed by Nanchung protein and Water witch protein.

[0139] Preferably, the candidate compound is a modulator that interferes with insect behavior and / or kills and / or repels insects.

[0140] In certain embodiments, the candidate compound binds directly to the complex formed by the Nanchung protein and the Water witch protein.

[0141] Preferably, the candidate compound modulates an ion channel formed by Water witch protein, or an ion channel formed by Nanchung protein and Water witch protein.

[0142] In certain embodiments, the Water Witch protein and / or the complex formed by the Nanchung protein and the Water Witch protein are present in biological cells. As used herein, the terms "biological cells" or "cells" have their commonly understood meanings. Within the cells, the Water Witch protein and / or the complex formed by the Nanchung protein and the Water Witch protein can be expressed from endogenous genes within the cells or from at least one vector transfected into the cells. It should be understood that either cultured cells, or cells obtained directly from animals such as frog oocytes, or whole tissues can be used as a source of the Water Witch protein and / or the complex formed by the Nanchung protein and the Water Witch protein. Preferably, the Nanchung and Water Witch proteins are co-expressed within the cells. The Nanchung and Water Witch proteins can be co-expressed in various ratios.

[0143] In certain embodiments, the Water Witch protein and / or the complex formed by the Nanchung protein and the Water Witch protein are formed in a cell-free system. As used herein, the term "cell-free system" refers to an in vitro system used to study biological reactions occurring within cells while reducing the complex interactions found in whole cells. Subcellular fractions can be isolated by ultracentrifugation to provide molecular machines that can be used in reactions in the absence of many other cellular components. Cell-free biological systems can be prepared by mixing one or more of the following: ribosomes, and / or DNA and / or RNA encoding one or more proteins of interest, membranes, and one or more enzymes and / or coenzymes. Cell-free biological systems have several advantages that make them suitable for industrial applications. In vitro biological systems are easily controlled. Cell-free systems offer an alternative option for accelerating protein synthesis, typically achieving very high product yields without the formation of by-products or the synthesis of cellular mass.

[0144] In certain embodiments, the cells are genetically modified to contain genes encoding one or more of the Nanchung and / or Water witch proteins.

[0145] The TRPV protein Nanchung and the TRPA protein Water witch can be used to generate ion channels for the purposes of the methods described herein.

[0146] In certain embodiments, nucleic acids encoding one or more TRPV proteins Nanchung and / or TRPA proteins Water witch are introduced into frog egg cells or other cells (parent cells) to produce TRPA and / or TRPV channel complexes for the methods described herein.

[0147] The insect TRPV and / or TRPA protein complexes and / or channels used in the screening assays of the present invention can be any insect TRPV and / or TRPA protein complexes and / or channels, or their homologs or conservative variants.For example, the TRPV and / or TRPA protein complexes and / or channels can be other members of the Drosophila TRPV and / or TRPA channel superfamily.Other sources of TRPV and / or TRPA protein complexes and / or channels can also be suitable for use in the methods of the present invention.

[0148] In some embodiments, the Nanchung nucleic acid sequence may be selected from the group consisting of: a) a polynucleotide molecule comprising a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NOs: 1-27, b) a polynucleotide molecule having at least about 70% sequence identity to a sequence of SEQ ID NOs: 1-27, and c) a fragment of the polynucleotide molecule of a) or b). Such a fragment may be a UTR, core promoter, intron, enhancer, cis-element, or other regulatory element.

[0149] In some embodiments, the Water witch nucleic acid sequence may be selected from the group consisting of: a) a polynucleotide molecule comprising a nucleic acid molecule having a sequence selected from the group consisting of SEQ ID NOs: 28-63, b) a polynucleotide molecule having at least about 70% sequence identity to a sequence of SEQ ID NOs: 28-63, and c) a fragment of the polynucleotide molecule of a) or b). Such a fragment may be a UTR, core promoter, intron, enhancer, cis-element, or other regulatory element.

[0150] The disclosed polynucleotides can provide for expression of Nanchung and Water witch proteins in a host cell.

[0151] In certain embodiments, the Nanchung protein sequence may be selected from the group consisting of: a) a polypeptide comprising an amino acid sequence having a sequence selected from the group consisting of SEQ ID NOs: 64-90; b) a polypeptide having at least 70% sequence identity to SEQ ID NOs: 64-90; and c) a fragment or conservative variant of the polypeptide of a) or b).

[0152] In certain embodiments, the Water witch protein sequence may be selected from the group consisting of: a) a polypeptide comprising an amino acid sequence having a sequence selected from the group consisting of SEQ ID NOs: 91-126, b) a polypeptide having at least 70% sequence identity to SEQ ID NOs: 91-126, and c) a fragment or conservative variant of the polypeptide of a) or b).

[0153] As used herein, "percent sequence identity" is determined by comparing two optimally aligned sequences over a comparison window, where the portion of the polynucleotide or polypeptide sequence within the comparison window may contain additions or deletions (i.e., gaps) compared to the reference sequence (which does not contain additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions where the same nucleic acid base or amino acid residue occurs in both sequences to determine the number of matching positions, dividing the number of matching positions by the total number of positions within the comparison window, and multiplying the result by 100 to determine the percentage of sequence identity. Alignment for determining percentage identity can be achieved in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms necessary to achieve optimal alignment across the entire length of the sequences being compared.

[0154] As used herein, the term "conservative variant" refers to an amino acid sequence in which a first amino acid is replaced with a second amino acid or amino acid analog having at least one similar biochemical property, which may be, for example, similar size, charge, hydrophobicity, or hydrogen-bonding capacity. For example, a first hydrophobic amino acid can be conservatively substituted with a second (non-identical) hydrophobic amino acid, such as alanine, valine, leucine, or isoleucine, or an analog thereof. Similarly, a first basic amino acid can be conservatively substituted with a second (non-identical) basic amino acid, such as arginine or lysine, or an analog thereof. Similarly, a first acidic amino acid can be conservatively substituted with a second (non-identical) acidic amino acid, such as aspartic acid or glutamic acid, or an analog thereof, or an aromatic amino acid, such as phenylalanine, can be conservatively substituted with a second aromatic amino acid or amino acid analog, e.g., tyrosine. In some embodiments, a peptide includes conservative variant substitutions of at least one amino acid, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acids. Typically, conservative variants will retain at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or more of the activity of the wild-type peptide sequence.

[0155] Exemplary conservative variant substitutions include replacement of alanine (A) with D-ala, Gly, Aib, β-Ala, Acp, L-Cys, or D-Cys; replacement of arginine (R) with D-Arg, Lys, D-Lys, homo-Arg, D-homo-Arg, Met, Be, D-Met, or D-Ile; replacement of asparagine (N) with D-Asn, Asp, D-Asp, Glu, D-Glu, Gln, or D-Gln; replacement of aspartic acid (D) with D-Asp, D-Asn, Asn, Glu, D-Glu, Gln, or D-Ile. or D-Gln; cysteine ​​(C) replaced by D-Cys, S-Me-Cys, Met, D-Met, Thr, or D-Thr; glutamine (Q) replaced by D-Gln, Asn, D-Asn, Glu, D-Glu, Asp, or D-Asp; glutamic acid (E) replaced by D-Glu, D-Asp, Asp, Asn, D-Asn, Gln, or D-Gln; glycine (G) replaced by Ala, D-Ala, Pro, D-Pro, Aib, β-Ala, or Acp; isoleucine (I) replaced by Replacement of leucine (L) by D-Leu, Val, D-Val, AdaA, AdaG, Leu, D-Leu, Met, or D-Met; replacement of lysine (K) by D-Lys, Arg, D-Arg, homo-Arg, D-homo-Arg, Met, D-Met, Ile, D-Ile, Orn, or D-Orn; replacement of methionine (M) by D-Met, S-Me-Cys, Be, D-Ile, Leu, D- replacement of phenylalanine (F) with D-Phe, Tyr, D-Thr, L-Dopa, His, D-His, Trp, D-Trp, Trans-3,4 or 5-phenylproline, AdaA, AdaG, cis-3,4 or 5-phenylproline, Bpa, or D-Bpa; replacement of proline (P) with D-Pro, L-thioazolidine-4-carboxylic acid, or D- or -L-1-oxazolidine-4-carboxylic acid (U.S. Pat. No. 4,511,390);These include, but are not limited to, replacement of serine (S) with D-Ser, Thr, D-Thr, allo-Thr, Met, D-Met, Met(O), D-Met(O), L-Cys, or D-Cys; replacement of threonine (T) with D-Thr, Ser, D-Ser, allo-Thr, Met, D-Met, Met(O), D-Met(O), Val, or D-Val; replacement of tyrosine (Y) with D-Tyr, Phe, D-Phe, L-Dopa, His, or D-His; and replacement of valine (V) with D-Val, Leu, D-Leu, Ile, D-Ile, Met, D-Met, AdaA, or AdaG;

[0156] Conservative variants of insect or mammalian TRPV and / or TRPA channels can be prepared according to methods for modifying peptide sequences known in the art, including those that can be found in references summarizing such methods, such as Molecular Cloning: A Laboratory Manual, J. Sambrook, et al., eds., Second Edition, Cold Spring Harbor or Current Protocols in Molecular Biology, F.M.Ausubel, et al., eds., John Wiley & Sons, Inc. New York.

[0157] Conservative variants of TRPV and / or TRPA channels can also be generated by modifying nucleic acids encoding TRPV and / or TRPA polypeptides.

[0158] In certain embodiments, the screening method of the present invention can be high-throughput screening.High-throughput screening (HTS) refers to the method of scientific experimentation that uses robotics, data processing and control software, liquid handling equipment, and highly sensitive detectors.High-throughput screening or HTS allows researchers to rapidly carry out millions of biochemical, genetic, or pharmacological tests.HTS is well known in the art, and can be found in, for example, U.S. Patent No. 5,976,813, U.S. Patent No. 6,472,144, U.S. Patent No. 6,692,856, U.S. Patent No. 6,824,982, and U.S. Patent No. 7,091,048.

[0159] HTS uses automation to perform assay screening against a library of candidate compounds. The assay is a test of specific activity, usually the inhibition or stimulation of a biochemical or biological mechanism. Specifically, the assay screens for the inhibition or stimulation of insect TRPV channels. A typical HTS screening library or "deck" can contain over 100,000 to 2,000,000 compounds.

[0160] The primary laboratory instrument or test vessel for HTS is the microtiter plate. This is a small, usually disposable, plastic container featuring a grid of small open divots called wells. Modern microplates for HTS typically have 384, 1536, or 3456 wells. These are all multiples of 96, reflecting the original 96-well microplate with 8 x 12 9 mm spaced wells.

[0161] To prepare for an assay, researchers fill each well of a plate with the appropriate reagent for the experiment they want to perform. After a period of incubation to allow the reagents to absorb, bind, or react (or not) with the compounds in the wells, measurements are taken across all the plate's wells, either manually or mechanically. Manual measurements are often necessary when researchers use a microscope to look for changes that (for example) cannot be easily determined by a computer alone. Otherwise, dedicated automated analyzers can perform many experiments on the wells, such as current or voltage measurements, colorimetric measurements, and radioactivity counts. In this case, the machine outputs the results of each experiment as a grid of numbers, each number mapped to the value obtained from a single well. High-volume analyzers can measure dozens of plates in this way in the space of a few minutes, generating thousands of experimental data points very quickly.

[0162] IV. Formulations, Uses and Applications In another aspect, the present invention provides compounds selected by the screening assays described herein. It is understood that analogs, derivatives, isomers, and pharmaceutically acceptable salts of the compounds selected by the screening assays described herein, as well as any formulations containing the selected compounds, are also included herein.

[0163] The compounds or groups of compounds determined or identified by the methods according to the invention can be used in methods of insect control, for example, by modulating the feeding behavior of insects. The identified compounds, their analogs, derivatives, isomers, and pharmaceutically acceptable salts are also referred to herein as "active agents" or "active ingredients."

[0164] Without being bound by theory, modulation of insect TRPV and / or TRPA channels induces signal transduction pathways that can kill, disrupt insect behavior, or repel insects. Thus, in some embodiments, the method includes modulating TRPV ion channels and / or TRPA channels with compounds identified by the screening methods described herein.

[0165] Thus, in certain embodiments, the present invention provides methods of insect control by killing, disrupting the behavior of, or repelling insects using compounds identified by the screening methods described herein. When used in the context of methods of insect control, compounds identified by the screening methods described herein also include analogs, derivatives, isomers, and pharmaceutically acceptable salts of such compounds.

[0166] In one embodiment, the methods and active agents described herein are applicable to insects that are agricultural or horticultural pests.

[0167] Examples of agricultural pests include Aphids (including Aphis fabae, Aphis gossypii, Aphis pomi, Aulacorthum solani, Brevicoryne brassicae, Dysaphis plantaginea, Macrosiphum euphorbiae, Macrosiphum euphorbiae, Macrosiphum rosae, Myzus persicae, Nasonovia ribisnigri); Whiteflies (Aleurodes spp., Bemisia spp., Dialeurodes spp., Trialeurodes spp.); Planthoppers (including Laodelphax striatellus, Nilaparvata lugens, Siphanta spp., Sogatella furcifera); Leafhoppers (including Amrasca spp., Empoasca spp.); Scales (Aspidiotus spp., Chrysomphalus aonidum, Icerya purchase, Unaspis citri); Mealybugs (including Maconellicoccus spp., Paracoccus spp., Planococcus spp.); Pollen beetles (including Carpophilus spp., Meligethes spp.); Thrips (including Caliothrips spp., Frankliniella spp., Scirtothrips spp., Thrips spp.); and Psyllids (including Bactericera spp., Cacopsylla spp., Diaphorina citri, Paratrioza cockerelli).

[0168] For example, vegetable and oilseed rape crops are susceptible to aphids, brown plant hoppers, alfalfa loopers, armyworms, beet armyworms, artichoke plume moths, cabbage budworms, cabbage loopers, cabbage webworms, corn earworms, celery leafeaters, cross-striped cabbageworms, European corn borers, diamondback moths, green cloverworms, invasive caterpillars, melonworms, omnivorous leafrollers, pyralid moths, and rindworms. The plant is susceptible to infestation by one or more of the following insect pests: soybean looper, tobacco budworm, tomato fruitworm, tomato hornworm, tomato pinworm, velvetbean caterpillar, and yellow-striped armyworm.

[0169] Similarly, grain feed and hay crops such as alfalfa, grass, and silage are often attacked by pests such as armyworms, beef armyworms, alfalfa caterpillars, European skippers, various inchworms and webworms, and yellow-striped armyworms.

[0170] Fruit and vine crops are susceptible to pests such as achema sphinx moth, amorbia, armyworms, citrus cutworm, blackheaded fireworm, blueberry leafroller, branchworm, cherry fruitworm, citrus cutworm, cranberry girdler, eastern tent caterpillar, fall webworm, fall webworm, hazel leafroller, hazel webworm, fruit leafroller, grape berry moth, grape leaffolder, grape leaf skeletonizer, green fruitworm, Gummosos-batrachedra commosae, gypsy moth, hickory shuckworm, hornworm, inchworm, navel orangeworm, obliquebanded leafroller, orrinivorous leafroller, omnivorous looper, orange tortrix moth, orangedog, oriental fruit moth, pandemis leafroller, peach twig borer, pecan nut casebearer, redbanded leafroller, redhumped caterpillar, roughskinned cutworm, sableworm, spanworm, tent caterpillar caterpillar, Thecla Thecla Basilides (thecla-theclaIt is susceptible to attack and defoliation by the following insects: the cotton bollworm, the tortrix moth, the tufted apple budmoth, the variegated leafroller, the walnut caterpillar, the western tent caterpillar, and the yellowstriped armyworm.

[0171] Field crops such as canola / rapeseed, evening primrose, meadowfoam, corn (fodder corn, sweet corn, popcorn), cotton, hops, jojoba, peanuts, rice, safflower, small grains (barley, oats, rye, wheat, etc.), sorghum, soybean, sunflower, and tobacco are often infested by armyworms, Asian corn borer and other corn borers, banded sunflower moth, beet armyworm, bollworm, cabbage looper, corn rootworm (including southern and western varieties), cotton leaf perforator, diamondback moth, European corn borer, and other insects. It is subject to infestation by insects including the soybean borer, green cloverworm, headmoth, headworm, invasive caterpillars, looper bugs (including Anacamptodes spp.), oblique banded leaf roller, omnivorous leaftier, podworin, podworm, fall webworm, southwestern corn borer, soybean looper, spotted cutworm, sunflower moth, cotton bollworm larvae, tobacco hornworm larvae, and noctuid larvae. Bedding plants, flowers, ornamentals, vegetables, and container stock are often food sources for numerous insect pests, such as armyworms, azalea moths, beet armyworms, diamondback moths, ello moths (hawkmoth larvae), Florida fern caterpillars, oleander moths, omnivorous leafrollers, omnivorous loopers, and tobacco.Forest, fruit, ornamental, and nut-bearing trees, as well as shrubs and other seedlings, are often infested with bagworms, blackheaded budworm, white-winged tussock moth, California oakworm, Douglas fir tussock moth, elm spanworm, fall webworm, fruit tree leafroller, greenstriped mapleworm, gypsy moth, jack pine caterpillar, mimosa webworm, pine butterfly, redhumped caterpillar, saddleback caterpillar, saddle prominent caterpillar, and other insects. They are susceptible to attack from a variety of pests, including spruce caterpillars, spring and fall cankerworms, spruce budworms, tent caterpillars, tortrix moths, and western tussock moths.

[0172] Similarly, turfgrass is often attacked by pests such as armyworms, sod webworms, and tropical sod webworms.

[0173] It is envisioned that the methods described herein are also applicable to pest control where the pest is not an insect, but rather is, for example, a nematode, slug, or snail.

[0174] In one embodiment, the methods described herein are also applicable to insect parasites. Some examples of insect parasites include Braconidae, family Braconidae; Ichneumonid Wasps, family Ichneumonidae; Chalcid Wasps, family Chalcidae; and Tachinid Flies, family Tachonidae.

[0175] In certain other embodiments, the methods described herein may also be applicable to insects that are disease vectors. Vectors are organisms that can introduce pathogens, such as bacteria or viruses, into a host organism, causing infection or disease. Exemplary disease vectors include, but are not limited to, mosquitoes, ticks, Siphonaptera (fleas), Diptera (flies), Phthiraptera (lice), and Hemiptera (true bugs).

[0176] Once a compound suitable for insecticidal use has been identified, the active ingredient or a formulation containing it may be applied directly to the target insect (i.e., larvae, pupae, and / or adults) or to the locus of the insect. In one embodiment, the active ingredient or a formulation and / or composition containing the active ingredient is applied directly to adult insects. In one embodiment, the active agent is applied directly to larvae and / or pupae of the target insect. In another embodiment, the active ingredient is applied to the locus of the insect.

[0177] Active agents can be conjugated to hydrophobic moieties, including but not limited to lipids and sterols, such that compounds incorporating such moieties penetrate the insect cuticle.

[0178] In one embodiment, the active ingredient or a formulation containing the active ingredient can be applied as a spray.For example, the active ingredient can be applied as an agricultural spray in aerial pesticide application, an environmental spray for controlling stinging insects, or a topical spray for localized control of biting insects.The active ingredient can also be formulated for spray application, such as an aerosol formulation.Spray application can be achieved using a spray pump.The active ingredient can also be encapsulated in materials such as starch, flour, and gluten in granular formulations.

[0179] In certain embodiments, the active ingredient or a formulation containing the active ingredient, when used for agricultural and / or environmental insect control, can be applied in conjunction with other insecticides and / or pesticides, such as organophosphates, synthetic pyrethroids, carbamates, chlorinated hydrocarbons, etc.

[0180] The active ingredient may be administered in an amount effective to induce the desired response, which is determined by routine testing.The actual effective amount will naturally vary depending on the specific active ingredient, the target insect and its developmental stage, application technique, the desired effect, and the duration of effect, and can be easily determined by those skilled in the art.The "effective amount of active ingredient" refers to the amount of active ingredient that regulates (activates or inhibits) insect TRPV channel and / or TRPA channel, for example, kills insects, interferes with insect behavior, or repels insects, thereby achieving the desired insect control.

[0181] Formulation methods are well known to those skilled in the art and can be found in Knowles, DA (1998) Chemistry and technology of agricultural formulations. Kluwer Academic, London (incorporated herein in its entirety by reference). Of course, those skilled in the art will recognize that formulation and application mode can affect the activity of the active ingredient in a given application. Therefore, for agricultural and / or horticultural use, TRPV inhibitors and / or agonists can be formulated as granules with a relatively large particle size (e.g., 8 / 16 or 4 / 8 US mesh), as water-soluble or water-dispersible granules, as powdery dust, as wettable powder, as emulsifiable concentrate, as aqueous emulsion, as solution, as suspension concentrate, as capsule suspension, as soluble (liquid) concentrate, as soluble powder, or any other known type of agriculturally useful formulation, depending on the desired application mode. It should be understood that the amounts specified herein are only intended to be approximate, as if the term "about" were placed before the specified amount.

[0182] These formulations can be applied as either water-diluted sprays, or dusts, or granules in the area where insect control is desired. The formulations may contain as little as 0.1%, 0.2%, or 0.5% by weight of the active ingredient, e.g., an insect TRPV inhibitor, to as much as 95% or more by weight.

[0183] "Dusts" are free-flowing mixtures of active ingredients with finely divided solids, such as talc, natural clay, diatomaceous earth, powders such as walnut shells and cottonseed flour, and other organic and inorganic solids that act as dispersants and carriers for toxicants, and these finely divided solids have an average particle size of less than about 50 microns. Typical dust formulations useful herein contain 90 parts, 80 parts, 70 parts, 60 parts, 50 parts, 40 parts, 30 parts, 20 parts, preferably 10 parts or less of the active ingredient, e.g., insect TRPV modulator or insect TRPV modulator. In one embodiment, the dust formulation may contain 1 part or less of the active ingredient and 99 parts or more of talc.

[0184] Wettable powders useful in formulations are in the form of finely divided particles that disperse easily in water or other dispersing agents. They are ultimately applied where insect control is needed, either as a dry dust or as an emulsion in water or other liquid. Typical carriers for wettable powders include fuller's earth, kaolin clay, silica, and other highly absorbent, easily wetted inorganic diluents. Wettable powders are typically formulated to contain approximately 5-80% active ingredient, depending on the absorbency of the carrier, and usually contain small amounts of wetting agents, dispersing agents, or emulsifiers to facilitate dispersion. For example, a useful wettable powder formulation contains 80.0 parts active ingredient, 17.9 parts palmetto clay, 1.0 part sodium lignosulfonate, and 0.3 parts sulfonated aliphatic polyester as a wetting agent. Additional wetting agents and / or oils may be added to the tank mix to facilitate dispersion on plant foliage.

[0185] Other useful formulations are emulsifiable concentrates (ECs), which are homogeneous liquid compositions dispersible in water or other dispersing liquids. They may consist entirely of the active ingredient and a liquid or solid emulsifier, or may also contain a liquid carrier such as xylene, heavy aromatic naphtha, isophorone, or other non-volatile organic solvent. For insecticide applications, these concentrates are dispersed in water or other liquid carrier and typically applied as a spray to the area to be treated. The weight percent of the essential active ingredient may vary depending on the manner in which the composition is applied, but generally, the insecticide composition contains 0.5 to 95 weight percent of the active ingredient.

[0186] Flowable formulations are similar to ECs, except that the active ingredient is suspended in a liquid carrier, typically water. Flowable formulations such as ECs may contain small amounts of surfactants and typically contain active ingredient in the range of 0.5 to 95% by weight, often 10 to 50% by weight, of the composition. For insecticide applications, flowable formulations may be diluted in water or other liquid vehicle and are typically applied as a spray.

[0187] Typical wetting agents, dispersants, or emulsifiers used in agricultural and / or horticultural formulations may include, but are not limited to, alkyl and alkylaryl sulfonates and sulfates and their sodium salts; alkylaryl polyether alcohols; sulfated higher alcohols; polyethylene oxide; sulfonated animal and vegetable oils; sulfonated petroleum oils; fatty acid esters of polyhydric alcohols and ethylene oxide addition products of such esters; and long-chain mercaptans and ethylene oxide addition products. Many other types of useful surfactants are commercially available. When used, surfactants typically comprise 1 to 15% by weight of the composition.

[0188] Other useful formulations include suspensions of the active ingredient in a relatively non-volatile solvent such as water, corn oil, kerosene, propylene glycol, or other suitable solvent.

[0189] In certain embodiments, formulations for pesticide applications may include a simple solution of the active ingredient in a solvent in which it is completely soluble at the desired concentration, such as acetone, alkylated naphthalenes, xylene, or other organic solvents.

[0190] Granular formulations, which have the active ingredient on relatively coarse particles, are particularly useful for aerial application or for penetration of cover crop canopies. Pressurized sprays, typically aerosols, in which the active ingredient is dispersed in finely divided form as a result of evaporation of a low-boiling point dispersing solvent carrier, may also be used. Water-soluble or water-dispersible granules are free-flowing, dust-free, and readily water-soluble or water-miscible. For use by farmers in the field, granular formulations, emulsifiable concentrates, flowable concentrates, aqueous emulsions, solutions, etc., may be diluted with water to give active ingredient concentrations ranging from approximately 0.1% or 0.2% to 1.5% or 2%.

[0191] The most frequently used formulations are water-miscible formulations that are mixed with water and then applied as a spray.Older water-miscible formulations include emulsifiable concentrates, wettable powders, water-soluble (liquid) concentrates, and soluble powders.Newer non-powder formulations, with reduced or no harmful solvents and improved stability, include suspension concentrates, capsule suspensions, and water-dispersible granules.Such formulations are preferably solutions and suspensions, such as aqueous suspensions and solutions, ethanolic suspensions and solutions, aqueous / ethanolic suspensions and solutions, saline solutions, and colloidal suspensions.

[0192] Alternatively, sprayable wax emulsion formulations may be used. The formulations contain the active ingredient in an amount of about 0.01% to 75% by weight. Aqueous wax emulsions are broadly described in U.S. Patent No. 6,001,346 (incorporated herein by reference in its entirety). The TRPV and / or TRPA modulators of the methods described herein may have a viscosity suitable for use in aerial spray or backpack spray applications.

[0193] The biodegradable wax carrier comprises at least about 10% by weight of the formulation. The biodegradable wax carrier is selected from the group consisting of paraffin, beeswax, vegetable waxes such as soybean wax (soybean-based), and hydrocarbon waxes such as Gulf Wax Household paraffin wax; paraffin wax, average melting point 53C (hexacosane), high molecular weight hydrocarbon), carnauba wax, lanolin, shellac wax, bayberry wax, sugarcane wax, microcrystalline, ozokerite, ceresin, montan, candelilla wax, and combinations thereof.

[0194] The formulation may also contain an emulsifier in an amount of about 1% to about 10% by weight. Suitable emulsifiers include lecithin and modified lecithin, mono- and diglycerides, sorbitan monopalmitate, sorbitan monooleate, sorbitan monolaurate, polyoxyethylene sorbitan monooleate, fatty acids, lipids, and the like. The emulsifier provides or improves the emulsifying properties of the composition. The emulsifier can be selected from a number of products well known in the art, including, but not limited to, sorbitan monolaurate (anhydrous sorbitol stearate, molecular formula C). 24 H 46O6), ARLACEL 60, ARMOTAN MS, CRILL 3, CRILL K3, DREWSORB 60, DURTAN 60, EMSORB 2505, GLYCOMUL S, HODAG SMS, IONET S 60, LIPOSORB S, LIPOSORB S-20, MONTANE 60, MS 33, MS33F, NEWCOL 60, NIKKOL SS 30, NISSAN NONION SP 60, NONION SP 60, NONION SP 60R, RIKEMAL S 250, sorbitan c, sorbitan stearate, SORBON 60, SORGEN 50, SPAN 55, and SPAN 60. Other sorbitan fatty acid esters that may be used include sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan sesquioleate, sorbitan trioleate, sorbitan monooleate, sorbitan trioleate.

[0195] In certain embodiments, the formulation can include a phagostimulant, such as corn oil, molasses, glycerol, or corn syrup, a proteinaceous material (protein or hydrolyzed protein), a sugar such as sucrose, or a food-based component, such as trimethylamine, putrescine, bacterial or yeast volatiles or metabolites, ammonium acetate, ammonium carbonate, or other ammonia-releasing compounds. The acetic acid vapor can be provided by compounds that produce volatile acetic acid, such as aqueous acetic acid, glacial acetic acid, glacial (concentrated) acetic acid, or ammonia-producing compounds, such as, but not limited to, ammonium hydroxide, ammonium carbonate, ammonium bicarbonate, ammonium acetate, etc.

[0196] In certain embodiments, the active ingredient may be formulated and / or applied with one or more secondary compounds. For example, various combinations of TRPV modulators and / or TRPA modulators can be used to obtain greater benefits. For example, both TRPV modulators and TRPA modulators may be applied simultaneously. Thus, in one embodiment, the formulations described herein may contain both TRPV modulators and TRPA modulators. In one embodiment, two or more active agents may be formulated together. In alternative embodiments, the two or more active agents formulated together are all TRPA modulators or all TRPV modulators. Such combinations can provide certain advantages, including, but not limited to, synergistic effects for greater insect or non-insect pest control, reduced application rates, thereby minimizing any impact on the environment and worker safety, and controlling a wider range of insect and non-insect pests and improving tolerance by non-pest species such as mammals and fish. Other second compounds include, but are not limited to, insecticides, pesticides, plant growth regulators, fertilizers, soil conditioners, or other agricultural and horticultural chemicals. The formulations may contain such second compounds in an amount of about 0.002% to about 25% by weight of the composition.

[0197] In certain embodiments, the formulations of the present invention may contain a visual attractant, such as a food colorant.

[0198] Various additives may also be incorporated into the formulation. These additives typically modify and / or enhance the physical characteristics of the carrier material, making it possible to tailor compositions with specific requirements, such as the release rate and amount of the active ingredient, protection of the wax composition from climatic conditions, etc. These additives are typically added in amounts of about 0.001% to about 10% by weight, more typically 1 to 6% by weight, and include, among others, plasticizers, volatilization inhibitors, antioxidants, lipids, various UV-blocking agents and absorbers, or antimicrobial agents.

[0199] Plasticizers, such as glycerin or soybean oil, can affect the physical properties of the composition and enhance its resistance to environmental damage.

[0200] Antioxidants, such as vitamin E, BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), and other antioxidants that protect bioactive agents from degradation, may be added in amounts of about 0.1% to about 3% by weight.

[0201] UV blockers, such as β-carotene, lignin, or p-aminobenzoic acid, protect the bioactive agent from photodegradation and can be added in amounts of about 1% to about 3% by weight.

[0202] Antimicrobial agents such as potassium sorbate, nitrates, nitrites, and propylene oxide protect the bioactive agents from destruction by microorganisms and may be added in amounts of about 0.1% to about 2% by weight.

[0203] Adjuvants can also be added to the formulation. An "adjuvant" is broadly defined as any substance added to the spray tank separate from the pesticide formulation that improves the performance of the pesticide. These can include, but are not limited to, wetting agents, spray agents, penetrants, compatibilizers, buffers, etc.

[0204] Other compounds and materials can also be added, provided they do not substantially interfere with the activity of the active ingredient. Whether an additive substantially interferes with the activity of the active ingredient can be determined by a standard test format, which involves directly comparing the efficacy of a composition of the active ingredient to which the compound has been added with the efficacy of a composition of the active ingredient to which the compound has been added.

[0205] It has been discovered that insects stop eating after ingesting TRPV channel activating compounds.For example, ingesting TRPV and / or TRPA ion-gated channel agonists can cause insects to stop eating.Therefore, in one embodiment, the compound can be formulated with insect food sources, for example, can be formulated with the compound in insect food.In another embodiment, the compound can be formulated with sucrose.Without wishing to be bound by theory, insects will feed on this mixture and stop eating.

[0206] In certain embodiments, the active agent may be applied to the feeding site of insects. This inhibits insect feeding and leads to starvation of insects. In one embodiment, the active agent is applied as a spray to the location where insects are present, for example, to the feeding site. The active agent can be applied to insect and / or pest (mollusk, nematode and / or arthropod) feeding sources, used in bait or traps, and / or applied directly to the pest itself.

[0207] In one embodiment, the active agent may be applied to the trap, for example, the trap may be coated with the active agent or the trap may contain insect food containing the active agent.

[0208] The nucleic acid sequences of the Nanchung and Water witch subunits of TRP channels, as well as the corresponding amino acid sequences, their origins, accession numbers (if available), and host organism codes, are summarized in Table 1. When Nanchung and Water witch proteins are co-expressed in cells, functional TRP channels can be formed. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0209] Adenovirions containing the adenoviral expression vectors of the present invention can be produced using the following methodology: Adenovirons can be used to generate cells that are used to express a protein of interest, e.g., Nanchung and / or Water witch proteins.

[0210] Methods for producing adenovirus generally involve introducing a vector containing a gene of interest (e.g., Nanchung or Water witch) into producer cells. The vector containing the gene of interest may be introduced into the producer cells using standard transfection techniques known to those skilled in the art (Zoltukhin et al., Gene Therapy, 6:973-985, 1999). The adenovirus produced by the producer cells (packaging cells) is used to transduce cells of interest. The cells of interest produce the protein of interest.

[0211] Preferably, the adenovirions are added to the cells at an appropriate multiplicity of infection according to standard transduction methods appropriate for the particular target cells. The titer of adenovirions administered can vary depending on the target cell type and the particular viral vector, and can be determined by one of skill in the art without undue experimentation.

[0212] The cell line may be an insect cell line, such as Sf9 (ATCC No. CRL-1711) cells or Schneider 2 (S2) cells (Life Technologies, No. R690-07) (transfection techniques, among others, may be used to introduce the gene of interest into the insect cells, if necessary), African clawed frog (Xenopus laevis) oocytes (injection techniques may be used to introduce the gene of interest), or a mammalian cell line, such as a mouse, hamster, or human cell line (several techniques may be used to introduce the gene of interest into mammalian cells, including, but not limited to, transfection, adenovirus, retrovirus, and / or electroporation), or any other cell line that does not normally express Nanchung and Water witch proteins. One example of a mammalian cell line suitable for use with the vector expression system of the present invention includes Chinese hamster ovary (CHO-K1) cells (ATCC No. CCL-61).

[0213] In some embodiments, cells expressing recombinant nucleic acid sequences encoding TRPV and / or TRPA channels are cells transformed with an expression vector containing a nucleotide sequence encoding an insect TRPV and / or TRPA channel (such as, but not limited to, the TRP channel proteins disclosed herein). Methods for transforming cells that would be known to those skilled in the art include, but are not limited to, infection using a viral vector, lipofection, electroporation, biolistics, and transfection. Detailed procedures for representative methods are described in Sambrook & Russell, 2001, and the references cited therein. Useful expression vectors and methods for introducing such vectors into cells or expressing the encoded polypeptides are also known to those skilled in the art. For example, plasmid expression vectors can be introduced into cells by calcium-phosphate-mediated transfection, DEAE-dextran-mediated transfection, lipofection, polybrene- or polylysine-mediated transfection, electroporation, or by conjugation to antibodies, gramicidin S, artificial viral envelopes, or other intracellular carriers. Viral expression vectors can be introduced into cells in an expressible form by, for example, infection or transduction, or by encapsulation in liposomes.

[0214] Once cells expressing recombinant nucleic acid sequences encoding insect TRPV and / or TRPA channel gene products have been produced, these cells can then be used to test candidate compounds for their ability to modulate ion transport within cells via TRPA and / or TRPV channels. Exemplary methods for testing ion transport within cells are described above and presented in the Examples section below. Other applicable methods will be known to those skilled in the art in light of the present disclosure. [Example]

[0215] The following examples provide exemplary embodiments. In light of the present disclosure and the general level of skill of those skilled in the art, those skilled in the art will understand that the following examples are intended to be illustrative only, and that numerous changes, modifications and alterations can be used without departing from the scope of the subject matter of the present disclosure.

[0216] Example I: Adenovirus expression constructs. To test the biological activity and ligand-binding properties of TRP channels, Drosophila melanogaster Nanchung, Inactive, and Water witch proteins were expressed alone or in combination in hamster CHO-K1 cells. To track TRP channel expression, they were fused with fluorescent proteins, AcGFP and mCherry, and antibody epitope tags, HA and FLAG, as shown in Figure 1. DNA encoding the tagged TRP channels was delivered into CHO-K1 cells using adenovirus. Adenovirus-mediated gene delivery allowed for expression of the gene of interest, optimization of subunit expression levels and stoichiometry, and prevention of potential toxic effects of insect TRP channels by maintaining virus-infected cells at room temperature.

[0217] To prevent TRP channel toxicity during the adenovirus packaging step, we used an adenovirus construct containing a Tet repressor binding site within a modified CMV promoter (Figure 1). The presence of the Tet repressor binding site inhibits expression of the gene of interest in adenovirus packaging cells that produce the Tet repressor.

[0218] Example II: Activation of TRP channels by afidopiropene and pymetrozine. The commercially available insecticides aphidopyropene (AP) and pymetrozine (PM) have previously been shown to activate and ultimately silence a complex consisting of Nanchung (Nan) and Inactive (Iav). Nanchung and Inactive are present in the insect chordotonal organ, the mechanosensor, which is involved in body coordination and sound and gravity sensing (Kavlie and Albert, 2013). Nanchung was shown to be the direct target of the insecticides, while Inactive was required for biological activity and high-affinity ligand binding (Kandasamy et al., 2017).

[0219] The TRPA channel Water witch (Water witch) is also present in chordotonal organs, and point mutations within a conserved region of Water witch, likely involved in ion conductance, produced effects similar to those of AP or PM (Senthilan et al., 2012). To test the hypothesis that Water witch may also be involved in the actions of PM and AP, we expressed Water witch alone or in combination with Nanchung in CHO-K1 cells. Given that TRP channels are permeable to Ca2+, we measured cellular responses by monitoring Ca2+ mobilization using the Ca2+-sensitive fluorescent probe FLUO4.

[0220] As shown in Figure 2, both PM and AP induced calcium mobilization in cells coexpressing Nanchung and Water witch (Figure 2B), but not in cells expressing Water witch individually (Figure 2A). It has previously been shown that Nanchung alone does not respond to stimulation with AP or PM, and coexpression of Inactive is required for biological activity. Therefore, the effects of coexpression of Nanchung and Water witch shown in Figure 2 are similar to those of coexpression of Nanchung and Inactive.

[0221] Notably, two other insecticidal compounds, flonicamid (FL) and TFNA-AM, which act on chordotonal organs but do not modulate insect TRPV channels, were ineffective.

[0222] Example III: Nanchung and Water witch are assembled into a Nanchung:Water witch complex. The observation that AP- and PM-induced calcium mobilization required coexpression of Nanchung and Water witch suggested that these two proteins might assemble into a Nanchung:Water witch complex. To test this hypothesis, Nanchung and Water witch were fused with different antibody epitope tags. As shown in Figure 3, HA-tagged Nanchung coimmunoprecipitated with FLAG-tagged Water witch. This is similar to the previously reported association of Nanchung and Inactive ...

[0223] Example IV: The Nanchung-Water witch complex is activated by the endogenous agonist nicotinamide. Nicotinamide, a form of vitamin B3, has been shown to activate both Nanchung and Inactive in insects and nematodes (Upadhyay et al., 2016). The stimulatory effect of nicotinamide required coexpression of Nanchung and Inactive. To test whether nicotinamide could also activate the Nanchung:Water witch complex, Nanchung, Inactive, and Water witch were expressed individually or in combination in CHO-K1 cells. As shown in Figure 4A, nicotinamide induced Ca2+ responses in cells coexpressing Nanchung and Water witch, as well as in cells coexpressing Nanchung and Inactive, but not in cells expressing these proteins individually. Furthermore, the nicotinamide-activated Nanchung:Inactive and Nanchung:Water witch complexes exhibited nearly identical potency and efficacy (Figure 4B). Thus, the Nanchung:Water witch complex can be activated by the same endogenous and exogenous modulators that activate the Nanchung:Inactive complex.

[0224] Example V: Co-expression of Nanchung and Water witch promotes binding of afidopiropene to Nanchung. As shown in Figure 5A, the specific binding of tritium-labeled aphidopyropene to membranes prepared from CHO-K1 cells expressing Nanchung was observed with a relatively moderate affinity (K dThis revealed the presence of a binding site with a Kd of 4.5 nM. Coexpression of Nanchung and Water witch increased the binding affinity by approximately 100-fold (Kd = 0.048 nM). Scatchard transformation of the binding isotherm (Figure 5B) further demonstrates the emergence of high-affinity binding sites upon coexpression of Water witch and Nanchung. Binding of tritiated AP to membranes expressing only Water witch was negligible. Therefore, similar to the previously reported increase in binding affinity upon coexpression of Nanchung and Inactive (Kandasamy et al., 2017), coexpression of Nanchung and Water witch dramatically enhances the binding of aphidpiropent.

[0225] Example VI: Pymetrozine and nicotinamide compete with afidopiropene for binding. As shown in Figure 6, both pymetrozine (Figure 6A) and nicotinamide (Figure 6B) completely inhibited the binding of tritiated afidopyropene to membranes prepared from cells co-expressing Nanchung and Inactive, as well as from cells co-expressing Nanchung and Water witch. These data indicate that afidopyropene, pymetrozine, and nicotinamide have overlapping binding sites, suggesting that the agonist-binding pockets of the NanchungInactive complex and the Water witchInactive complex may be structurally similar.

[0226] Example VII: Water witch forms functional channels activated by plant-derived blockers. The biological activity of the plant-derived inhibitors and / or pesticides allyl isothiocyanate and cinnamaldehyde has been attributed solely to the activation of the insect TRPA1 channel. Based on amino acid sequence analysis, the TRPA1 and Water Witch proteins were confirmed to belong to distinct clades, the TRPA1 clade and the basal TRPA clade, respectively. Unlike the TRPA1 clade, members of the basal TRPA clade were predicted to be inefficient at detecting electrophilic compounds such as allyl isothiocyanate and cinnamaldehyde (Kang et al., 2010). However, we found that both Water Witch expressed individually and Water Witch co-expressed with Nanchung could be efficiently activated by allyl isothiocyanate and cinnamaldehyde (Figure 7). The Nanchung:Inactive complex did not respond to these compounds. This finding identifies Water witch protein as a new alternative target for plant-derived deterrent and repellent agents and suggests that assays utilizing heterologously expressed Water witch (assys) can be used to discover new compounds with deterrent and / or repellent properties.

[0227] Example 8. The similarities between Water witch and Inactive are limited. The Inactive and Water Witch proteins belong to different subfamilies, the TRPV and TRPA subfamilies, respectively. Furthermore, amino acid alignment (Figure 8) revealed that the Inactive and Water Witch proteins of Drosopila melanogaster share only approximately 22% identity. Nevertheless, we demonstrated that the Water Witch protein can assemble with the Nanchung protein, conferring agonist responses and promoting aphidopylopene binding similarly to the Inactive protein (Kandasamy et al., 2017; Nesterov et al., 2015). Considering that the Nanchung:Inactive and Nanchung:Water Witch complexes interact with endogenous and exogenous agonists with nearly identical potency, efficacy, and affinity, it is possible that the ligand-binding pocket is located entirely within the Nanchung protein, while Water Witch or Inactive acts like a chaperone to facilitate the folding of Nanchung and / or alter its conformation. This raises the possibility that any other TRP channels that can form a complex with Nanchung may function similarly.

[0228] In summary, using a combination of functional and binding assays in the above examples, we have demonstrated the following: i) the insect proteins Nanchung and Water witch can assemble into a complex; ii) the Nanchung-Water witch complex is a direct target of the commercial insecticides afidopyropene and pymetrozine, as well as the endogenous agonist nicotinamide; iii) afidopyropene, pymetrozine, and nicotinamide have overlapping binding sites; iv) Nanchung protein forms the primary binding interface for afidopyropene, but the presence of Water witch protein increases the binding affinity; v) Water witch protein is a direct target for plant-derived insecticides and repellents; and vi) a new method has been developed that acts by directly targeting insect Water witch protein or the Nanchung-Water witch complex and can be used to identify candidate compounds that can be used as insecticides or repellents.

[0229] material and method compound Pymetrozine was obtained from Sigma, flonicamid was purchased from ChemService Inc. (West Chester, PA), and TFNA-AM (4-trifluoronicotinamide) was obtained from ABCR GmbH (Karlsruhe, Germany). Afidopyropene was synthesized at BASF. Tritium-labeled afidopyropene (19 Ci / mmol) was custom-made by RC TRITEC AG (Teufen, Germany).

[0230] Heterogeneous manifestations of Nanchung, Inactive, and Water witch To generate expression constructs, complementary DNA encoding Drosophila melanogaster Water witch (SEQ ID NO: 63) and Inactive (NP_572353) was fused in-frame at its carboxyl terminus with a FLAG tag and AcGFP fluorescent protein (Figure 1). Complementary DNA encoding Drosophila melanogaster Nanchung (SEQ ID NO: 7) was fused in-frame at its carboxyl terminus with an HA tag and mCherry fluorescent protein (Figure 1). The complementary DNA was codon-optimized for expression in mammalian cells.

[0231] To produce adenovirus, complementary DNA encoding the epitope-tagged TRP channel was subcloned into the adenovirus shuttle vector pENTCMV1-TetO (Welgen Inc., Worceter, MA) and ligated into the pAdREP plasmid (Welgen), which contains the remaining adenovirus genome. The pENTCMV1-TetO vector contains two Tet repressor binding sites within a modified CMV promoter, which repress transcription of the gene of interest in cells expressing the Tet repressor (Figure 1). The recombinant product was transformed into Escherichia coli cells, positive clones were selected, and cosmid DNA was isolated. Transfection of the cosmid DNA into HEK293-TetR cells resulted in the production of the Tet repressor (Postle et al., 1984), preventing TRP channel expression by the adenovirus-packaging cells. Adenovirus was purified from a large-scale culture, and the viral titer was 260 nm × 1.1 × 10 14 was calculated as the absorption at

[0232] Calcium imaging Hamster CHO-K1 cells (ATCC® CCL-61™) were infected with adenoviruses expressing TRP channels individually or in combination. Cells were maintained at 37°C overnight, followed by 3 days at 25°C. The medium was changed two days after seeding. Calcium mobilization was detected using the fluorescent calcium probe fluo-4AM (Life Technologies) and measured with a FLIPR-TETRA instrument (Molecular Devices, Sunnyvale, CA). Cells were loaded with 50 μl of Hank's balanced salt solution (HBSS) containing 4 μM fluo-4AM, 5 mM probenecid, 20 mM CaCl2, and 0.02% pluronic acid for 2 hours at 25°C. The dye was then discarded, and 50 μl of HBSS was added. Test compounds were dissolved in DMSO and added to the cells in 50 μl of HBSS to obtain a final DMSO concentration of 0.2%. Fluorescence was monitored at excitation / emission wavelengths of 470–495 nm / 515–575 nm at 1-second intervals for 10 minutes. 50 Values ​​were calculated using GraphPad Prism (GraphPad Software).

[0233] Membrane preparation Adenovirus-transduced CHO-K1 cells were cultured at 10 × 10 per T175 flask (Greiner). 6Cells were seeded and maintained at 37°C for 18 hours, followed by incubation at 25°C for 72 hours. The medium was changed 24 and 72 hours after transduction. Cells were rinsed twice with PBS and incubated with Versene solution (Life Technologies) for 10 minutes at room temperature. Dissociated cells were pelleted at 300 × g for 10 minutes and resuspended in 8 mL of ice-cold 20 mM Hepes (pH 7.3) containing 1% (wt / vol) protease inhibitor mixture (Halt, Thermo Fisher Scientific). After 10 minutes on ice, cells were homogenized by sonication. The sample was centrifuged at 300 × g for 10 minutes, and the supernatant was collected and centrifuged at 50,000 × g for 2 hours. Membranes were resuspended in 0.5 mL of 50 mM HEPES, 100 mM NaCl, 1 mM CaCl2, 5 mM MgCl2, pH 7.4, and stored at -80°C. Protein content was determined using the Bradford method.

[0234] [ 3 H]-Afidopyropene binding assay Equilibrium binding assays were performed in binding buffer (50 mM HEPES, 100 mM NaCl, 1 mM CaCl, 5 mM MgCl, pH 7.4). Reactions were performed in 96-well U-bottom polypropylene plates. 100 μL of membranes resuspended in binding buffer containing Tween-20 (final concentration in the assay: 0.0025%) dissolved in ethanol [ 3The mixture was combined with 200 μl (saturation binding experiments) or 800 μl (ligand displacement experiments) of binding buffer containing [H]-afidopyropenem, and, where indicated, the test compound dissolved in DMSO. The final concentrations of ethanol and DMSO were kept at 0.2% and 0.25%, respectively, for both saturation binding and competition experiments. Typically, 60–100 μg of membranes were used per data point. After 3 h of incubation at room temperature, the assay mix was filtered through a MultiScreen HTS+96 Hi Flow FC filter (EMD Millipore) pre-wetted with 0.1% polyethyleneimine. The filters were washed three times with 200 μl of ice-cold binding buffer, scintillation cocktail was added, and radioactivity was measured using a Wallac 1450 Microbeta counter. Nonspecific binding was measured in the presence of a 500-fold excess of unlabeled afidopyropenem. i Values ​​were calculated by the Cheng-Prusoff method (Ref).

[0235] Western blotting and co-immunoprecipitation For Western blotting, adenovirus-transduced cells were seeded in 35-mm dishes. Cells were washed with phosphate-buffered saline and lysed in 300 μl of NuPAGE LDS sample buffer (Life Technologies) supplemented with protease inhibitor cocktail (Sigma-Aldrich) and TurboDNAse (Ambion). For co-immunoprecipitation, adenovirus-transduced cells were seeded in 90-mm dishes and lysed in 50 mM HEPES, 10% glycerol, 1% Triton-X-100, 100 mM NaCl, 1 mM EDTA, and 1 mM EGTA (containing 0.5 mM PMSF and protease inhibitor cocktail (Sigma)). FLAG-tagged Water witch proteins were immunoprecipitated with anti-FLAG M2 affinity gel (Sigma). Samples were electrophoresed using the NuPaGe 4-12% Bis-Tris Pre-Cast Gel System (Life Technologies). Proteins were detected using anti-FLAG M2-peroxidase antibody (Sigma) or anti-HA-biotin antibody (Covance). Blots were developed using ECL reagents (Thermo Scientific).

[0236] Nanchung and Water witch sequence alignments were performed by ClustalW. The transmembrane domains of Inactive and Water witch were assigned by homology with the low-temperature electronic structures of rat TRPV6 and human TRPA1, respectively, using molecular operating environment modeling software.

[0237] Accordingly, it is intended that the foregoing detailed description be considered illustrative rather than limiting, and it is understood that it is the following claims, including all equivalents, that are intended to define the spirit and scope of the invention. The present specification includes the following aspects. [1] 1. A method for determining whether a candidate compound is a modulator of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein, comprising: (a) providing a first cell that co-expresses the insect transient receptor potential channel Nanchung protein and the insect transient receptor potential channel Water witch protein; (b) contacting the first cell with the candidate compound; (c) assaying for modulation of said complex, wherein said modulation identifies said candidate compound as a modulator of said complex. [2] The assaying step comprises: (1) detecting calcium ion mobilization in the first cell in response to the candidate compound; or (2) detecting a membrane potential in the first cell in response to the candidate compound. [3] The assaying step comprises: (1) comparing calcium ion mobilization in the first cell in the absence of the candidate compound with calcium ion mobilization in the first cell in the presence of the candidate compound; or (2) comparing the membrane potential in the first cell in the absence of the candidate compound with the membrane potential in the first cell in the presence of the candidate compound. [4] The assaying step comprises: (1) comparing calcium ion mobilization in the first cell in the presence of the candidate compound with a reference level of calcium ion mobilization indicative of no modulation of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein; or (2) comparing the membrane potential in the first cell in the presence of the candidate compound with a membrane potential reference level indicating no modulation of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein. [5] The method according to any one of [2] to [4], wherein the candidate compound regulates the calcium ion mobilization or the membrane potential in the first cell by at least 20% relative to the reference level. [6] The method according to any one of [1] to [4], wherein the candidate compound is a modulator that inhibits the activity of the complex. [7] The method according to any one of [1] to [4], wherein the candidate compound is a modulator that activates the complex. [8] The method according to any one of [1] to [4], wherein modulation of the complex by the candidate compound disrupts insect behavior. [9] The method according to any one of [1] to [4], wherein modulation of the complex by the candidate compound kills the insect.

[10] The method according to any one of [1] to [4], wherein modulation of the complex by the candidate compound repels insects.

[11] The method according to any one of [1] to [4], wherein the candidate compound is selected from the group consisting of organic small molecules, inorganic small molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

[12] The method according to any one of [1] to [4], wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

[13] Specific species of lepidoptera include Achroia grisella, Acleris spp. (including A. fimbriana, A. gloverana, and A. variana); Acrolepiopsis assectella 、Acronita major 、Adoxophyes spp spp.(from A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp spp. (including A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyresthia conjugella, Argyroploce spp., Argyrotaenia spp. (including A. velutiNanchunga); Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp.、Cacoecia spp.(from C.muriNanchungaC.podana);Cactoblastis cactorum、Cadra cautella、Calingo braziliensis、Caloptilis theivora、Capua reticulana、Carposina spp aridula, Cheimatobia brumata, Chilo spp. (including C. Indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp spp. 、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus);Desmia funeralis, Diaphania spp.hyalinata);Diatraea grandiosella、Diatraea saccharalis、Diphthera festiva、Earias spp.(E.insulana、E.vittella);Ecdytolopha aurantianu、Egira(=Xylomyges)curialis、Elasmopalpus lignosellus、Eldana saccharina、Endopiza viteana、Ennomos subsignaria、Eoreuma loftini、Ephestia spp thrax、Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Faronta albilinea、Feltia spp mellonella、Gracillaria spp.、Grapholite spp.(G.funebrana、G.molesta、G.inopinata); spp. (including H. armigera (=Heliothis armigera) and H. zea (=Heliothis zea)); spp.(excluding H.undalis, H. spp.rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorella, Lerodea eufala, Leucinodes orbonalis, Leucoma salicis, Leucoptera spp. (e.g., L.coffeella, L.scitella); Leuminivora lycinivorell, Lithocolletis blancardella, Lithophane antennata, Llattia octo(=Amyna axis), Lobesia botrana, Lophocampa spp., Loxagrotis albicosta, Loxostege spp. (e.g., L.sticticalis, L.cereralis); Lymantria spp. (e.g., L.dispar, L.monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta);Marasmia spp.Marmara spp.Maruca testulalis、Megalopyge lanata、Melanchra picta、Melanitis leda、Mocis spp separate、Nemapogon cloacella、Neoleucinodes elegantalis、Nepytia spp.、Nymphula spp.、Oiketicus spp.、Omiodes indicata、Omphisa anastomosalis、Operophtera brumata、Orgyia pseudotsugata、Oria spp.Orthaga thyrisalis, Ostrinia spp. (O.nubilalis); Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp licorice, Paysandisia archon, Pectinophora spp., and P. gossypiella; Peridroma saucia, Perileucoptera spp., such as P. coffeella; Phalera bucephala, Phryganidia californica, and Phthorimaea spp. (including P. operculella);Phyllocnistis citrella; Phyllonorycter spp. (including P. blancardella, P. crataegella, P. issikii, P. ringoniella); Pieris spp.(such as P. brassicae, P. rapae, P. napi);Pilocrocis tripunctata, Platypena scabra, Platynota spp.stultana); Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp. (e.g., P.sequax, P.unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana,. Sabulodes aegrotata Schizura concinna Schoenobius spp. Schreckensteinia festaliella Scirpophaga spp spp.(S.inferens、Seudyra subflava、Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.(singえば、T.ni);Night(=Scrobipalpula)absolute、Udea spp.(singhえば、U.rubigalis、U.rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, such as Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A.anxius, A.planipennis, A.sinuatus); Agriotes spp. (e.g. A.fuscicollis, A.lineatus, A.obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A.glabripennis); Anthonomus spp. (e.g. A.eugenii, A.grandis, A.pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A.linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g., B.lentis, B.pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C.assimilis, C.napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E. similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp. (e.g., M. aeneus); Melolontha spp. (e.g., M. hippocastani, M. melolontha); Metamasius hemipterus, Microtheca spp., Migdolus spp. (e.g., M. fryanus), Monochamus spp. (e.g., M. alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P.helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R. billineatus, R. ferrugineus, R. palmarum, R. phoenicis, R. vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S. granaria, S. oryzae, S. zeamais); Sphenophorus spp. (e.g., S. levis); Stegobium paniceum, Sternechus spp. (e.g., S. subsignatus); Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp.、Tenebrio molitor、Tenebrioides mauretanicus、Tribolium spp.(for example、T.castaneum); spp.quadrimaculatus, A.sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. (e.g. C.bezziana, C.hominivorax, C.macellaria); Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp. (e.g. C.hominivorax); Contarinia spp. (e.g. C.sorghicola); Cordylobia anthropophaga, Culex spp. (e.g. 、 C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g. D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g. D. suzukii), Fannia spp. (e.g. F. canicularis); Gastraphilus spp. (e.g. G. intestinalis); Geomyza tipunctata, Glossina spp. (e.g. G. fuscipes, G. morsitans, G. palpalis, G. tachinoides); Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp. (e.g. H. platura); Hypoderma spp.(eg H.lineata);Hyppobosca spp.、Hydrellia philippina、Leptoconops torrens、Liriomyza spp.(eg L.sativae、L.trifolii);Lucilia spp.(eg L.caprina、L.cuprina、L.sericata);Lycoria pectoralis、Mansonia titillanus、Mayetiola spp.(eg M.destructor); spp. (for example, M.autumnalis, M. domestica); Muscina stablens, Oestrus spp. (for example, O. ovis);frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp.; Insects from the order Thysanoptera, e.g., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp. (e.g., F.fusca, F.occidentalis, F.tritici); Heliothrips spp., Hercinothrips femoralis, Kakothrips spp., Microcephalothrips abdominalis, Neohydatothrips samayunkur, Pezothrips kellyanus, Rhipiphorothrips cruentatus, Scirtothrips spp. (e.g., S.citri, S.dorsalis, S.perseae);Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequence、Thrips spp. (including T.imagines, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, and T.tabaci); spp.(such as A.hilare);Acyrthosipon spp.(such as A.onobrychis)A.pisum);Adelges laricis, Adelges tsugae, Adelphocoris spp.(such as A.rapidus,A.superbus);Aeneolamia spp. Agonoscena spp. Aulacorthum solani Aleurocanthus woglumi Aleurodes spp. Aleurodicus disperses Aleurolobus barodensis Aleurothrixus spp. Amrasca spp spp.、Anuraphis cardui、Aonidiella spp.、Aphanostigma pyri、Aphidula nasturtii、Aphis spp.(including A. craccivora, A. fabae, A. forbesi, A. gossypii, A. grossulariae, A. maidiradicis, A. pomi, A. sambuci, A. schneideri, A. spiraecola); Arboridia apicalis, Arilus critatus, Aspidiella spp., Aspidiotus spp., Atanus spp., Aulacaspis yasumatsui, Aulacorthum solani, Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp tabaci));Blissus spp.leucopterus; Brachycaudus spp. (e.g., B. cardui, B. helichrysi, B. persicae, B. prunicola); Brachycolus spp., Brachycorynella asparagi, Brevicoryne brassicae, Cacopsylla spp. (e.g., C. fulguralis, C. pyricola (Psylla piri); Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes ceriferus, Cerosipha gossypii, Chaetosiphon fragaefolii, Chionaspis tegalensis, Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila, Cimex spp. (e.g., C. hemipterus, C. lectularius); Coccomytilus halli, Coccus spp. (e.g., C. hesperidum, C. pseudomagnoliarum); Corythucha arcuata, Creontiades dilutus, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasynus piperis, Dialeurodes spp. (e.g., D. citrifolii); Dalbulus maidis, Diaphorina spp. (e.g., D. citri); Diaspis spp. (e.g., D. bromeliae); Dichelops furcatus, Diconocoris hewetti, Doralis spp.、Dreyfusia nordmannianae、Dreyfusia piceae、Drosicha spp.、Dysaphis spp.(eg D.plantaginea、D.pyri、D.radicola);Dysaulacorthum pseudosolani、Dysdercus spp.(eg D.cingulatus、D.intermedius);Dysmicoccus spp.、Edessa spp、Geocoris spp、Empoasca spp.(eg E.fabae、E.solana);Epidiaspis leperii、Eriosoma spp.(eg E.lanigerum、E.pyricola);Erythroneura spp.、Eurygaster spp.(eg E.integriceps);Euscelis bilobatus、Euschistus spp.(eg E.heros、E.impictiventris、E.servus);Fiorinia theae、Geococcus coffeae、Glycaspis brimblecombei、Halyomorpha spp.(eg H.halys);Heliopeltis spp.、Homalodisca vitripennis (=H.coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp. (e.g. I.purchase); Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp. (e.g. L.ulmi); Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp. (e.g. L.hesperus, L.lineolaris, L.pratensis);. Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp. (e.g. M.rosae, M.avenae, M.euphorbiae); Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (=Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis coryli, Murgantia spp., Myzus spp. (e.g. M.ascalonicus, M.cerasi, M.nicotianae, M.persicae, M.varians); Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp. (e.g. N.malayanus, N.nigropictus, N.parvus, N.virescens); Nezara spp.(eg N.viridula);Nilaparvata lugens、Nysius huttoni、Oebalus spp.(eg O.pugnax);Oncometopia spp.、Orthezia praelonga、Oxycaraenus hyalinipennis、Parabemisia myricae、Parlatoria spp.、Parthenolecanium spp.(eg、P.corni、P.persicae);Pemphigus spp.(eg、P.bursarius、P.populivenae);Peregrinus maidis、Perkinsiella saccharicida、Phenacoccus spp. (for example、P. aceris、P.gossypii); Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp. (e.g. P.devastatrix), Piesma quadrata, Piezodorus spp. (e.g. P.guildinii); Pinnaspis aspidistrae, Planococcus spp. (e.g. P.citri, P.ficus); Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudacysta persea, Pseudaulacaspis pentagona, Pseudoococcus spp. (e.g. P.comstocki); Psylla spp. (e.g. P.mali); Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp. (e.g. Q.perniciosus); Quesada gigas, Rastroccus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp. (e.g., R. pseudobrassicas, R. insertum, R. maidis, R. padi); Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulatus, Sitobion avenae, Sogata spp.、Sogatella furcifera、Solubea insularis、Spissistilus festinus(=Stictocephala festina); spp.(including T.accerra;T.perditor);Tibraca spp., Tomaspis spp., Toxoptera spp.(including T.aurantii);Trialeurodes spp.(including T.abutilonea, T.ricini, and T.vaporariorum);Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp.(including U.citri,U.yanonensis);Viteus vitifolii;The list of Hymenoptera includes Acanthomyops interjectus, Athalia rosae, Atta spp (including A. capiguara, A. cephalotes, A. cephalotes, A. laevigata, A. robusta, A. sexdens, A. texana), Bombus spp., Brachymyrmex spp., and Camponotus spp (from C. floridanus, C. pennsylvanicus, C. modoc); Cardiocondyla nuda, Chalibion ​​sp, Crematogaster spp, Dasymutilla occidentalis, Diprion spp, Dolichovespula maculata, Dorymyrmex spp, Dryocosmus kuriphilus, Formica spp, Hoplocampa spp., (excluding H.minuta, H. testudinea); Iridomyrmex humilis, Lasius spp.niger) Linepithema humile Liometopum spp Leptocybe invasa Monomorium spp(M.pharaonis) Monomorium Nylandria fulva Pachycondyla chinensis Paratrechina longicornis Paravespula spp(such as P. germanica, P. pennsylvanica, P. vulgaris);Pheidole spp(such as P. megacephala);Pogonomyrmex spp(such as P. barbatus, P. californicus), Polistes rubiginosa、Prenolepis impairs、Pseudomyrmex gracilis、Schelipron spp、Sirex cyaneus、Solenopsis spp(S.geminata、S.invicta、S.molesta、S.richteri、S.xyloni) Sphecius speciosus spp (including V.squamosal);Wasmannia auropunctata, Xylocopa sp.;Orthoptera species include Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, and Ceuthophilus spp. Diastrammena asynamora. Dociostaurus maroccanus spp.(such as L. migratoria, L. pardalina);Melanoplus spp.(such as M. spp.).bivittatus, M. femurrubrum, M. mexicanus, M. sanguinipes, M. spretus); spp(S.americana, S.gregaria), Stemopelmatus spp, Tachycines asynamorus, and Zonozerus variegatus;Arachnidae, Argasidae, Ixodidae, Sarcoptidae (Acari), Amblyomma spp.(A.americanum, A.variegatum, A.maculatum), Argas spp., (A. persicu), Boophilus spp., B. annulatus, B. decoloratus, B. microplus) Dermacentor spp (including D. silvarum, D. andersoni, D. variabilis), Hyalomma spp. (including H. truncatum), Ixodes spp.(I.ricinus, I.rubicundus, I.scapularis, I.holocyclus, I.pacificus) Rhipicephalus sanguineus Ornithodorus spp.(excluding O. moubata, O. hermsi, O. turicata), Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp., (excluding P.ovis), Rhipicephalus spp(including R.sanguineus, R.appendiculatus), Rhipicephalus evertsi), Rhizoglyphus spp; Sarcoptes spp. (including S. Scabiei); and Aceria spp (including A. sheldoni, A. anthocoptes), Acallitus spp; Aculops spp.pelekassi); Aculus spp. (e.g., A. schlechtendali); Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; the Eriophyidae family, including Eriophytes ribis and Eriophyes spp. (e.g., Eriophyes sheldoni); the Tarsonemiidae family, including Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp., and Stenotarsonemus spinki; the Tenuipalpidae family, including Brevipalpus spp. (e.g., B. phoenicis); Eotetranychus spp., Eut. The Tetranychidae family includes species such as etranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp. (e.g., T. cinnabarinus, T. evansi, T. kanzawai, T. pacificus, T. phaseulus, T. telarius, and T. urticae); Bryobia praetiosa; Panonychus spp. (e.g., P. ulmi, P. citri); Metatetranychus spp., and Oligonychus spp. (e.g., O. pratensis, O. perseae); Vasates lycopersici; Raoiella indica; Carpoglyphidae, including Carpoglyphus spp.; Penthaleidae spp. (e.g., Halotydeus destructor); Demodicidae, including species such as Demodex spp.; Trombicula family Trombicidea, including spp.; family Macronyssidae, including Ornothonyssus spp.; family Pyemotidae, including Pyemotes tritici; Tyrophagus putrescentiae; family Acaridae, including Acarus siro; family Araneida, including Latrodectus mactans, Tegenaria agrestis, Chiracanthium spp., Lycosa spp., Achaearanea tepidariorum, and Loxosceles reclusa; pests from the phylum Nematoda, for example, plant-parasitic nematodes (e.g., root-knot nematodes) Meloidogyne spp. (e.g., M. hapla, M. incognita, M. javanica); cyst nematodes Globodera spp. (e.g., G. rostochiensis); Heterodera (e.g. H. avenae, H. glycines, H. schachtii, H. trifolii); Seed gall nematodes Anguina spp.Stem and foliar nematodes Aphelenchoides spp. (e.g., A. besseyi); Sting nematodes Belonolaimus spp. (e.g., B. longicaudatus); Pine nematodes Bursaphelenchus spp. (e.g., B. lignicolus, B. xylophilus); Ring nematodes Criconema spp.; Criconemella spp. (e.g., C. xenoplax and C. ornata); and Criconemoides spp. (e.g., Criconemoides informis; Mesocriconema spp.); Stem and bulb nematodes Ditylenchus spp. (e.g., D. destructor, D. dipsaci); Awl nematodes (Dolichodorus spp.); spiral nematodes (Heliocotylenchus multicinctus); sheath and sheathoid nematodes (Hemicycliophora spp. and Hemicriconemoides spp.); Hirshmanniella spp.; lance nematodes (Hoploaimus spp.); false rootknot nematodes (Nacobbus spp.); needle nematodes (Longidorus spp. (e.g., L. elongatus); root-lesion nematodes (Pratylenchus spp. (e.g., P. brachyurus, P. neglectus, P. penetrans, P. curvitatus, P. goodeyi); Burrowing nematodes Radopholus spp. (e.g., R. similis; Rhadopholus spp.; Rhodopholus spp.;Reniform nematodes Rotylenchus spp. (e.g., R. robustus, R. reniformis); Scutellonema spp.;Stubby-root nematodes Trichodorus spp. (e.g., T. obtusus, T. primitivus); Paratrichodorus spp. (e.g., P. minor);Stunt nematodes Tylenchorhynchus spp. (e.g., T. claytoni, T. dubius);Citrus nematodes Tylenchulus spp. (e.g., T. semipenetrans);Dagger nematodes Xiphinema spp.;and other plant-parasitic nematode species;Insecta from the order Isoptera, for example, Calotermes flavicollis, Coptotermes spp (e.g. C. formosanus, C. gestroi, C. acinaciformis); Cornitermes cumulans, Cryptotermes spp (e.g. C. brevis, C. cavifrons); Globitermes sulfureus, Heterotermes spp (e.g. H. aureus, H. longiceps, H. tenuis); Leucotermes flavipes, Odontotermes spp., Incisitermes spp (e.g. I. minor, I. Snyder); Marginitermes hubbardi, Mastotermes spp (e.g. M. darwiniensis), Neocapritermes spp (e.g. N.opacus, N.parvus); Neotermes spp, Procornitermes spp, Zootermopsis spp. (e.g. Z. angusticollis, Z. nevadensis), Reticulitermes spp. (e.g. R. hesperus, R. tibialis, R. speratus, R. flavipes, R. grassei, R. lucifugus, R.santonensis, R.virginicus);Termes natalensis;Insects from the order Blattaria, such as Blatta spp (e.g. B.orientalis, B.lateralis);Blattella spp (e.g. B.asahinae, B.germanica);Leucophaea maderae, Panchlora nivea, Periplaneta spp (e.g. P.americana, P.australasiae, P.brunnea, P.fuligginosa, P.japonica); Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnoscelus surinamensis; insects from the order Siphonoptera, e.g. Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp (e.g. C. felis, C. canis), Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus; insects from the order Thysanura, for example, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica; pests from the class Chilopoda, for example, Geophilus spp., Scutigera spp. (e.g., Scutigera coleoptrata); pests from the class Diplopoda, for example, Blaniulus guttulatus, Julus spp., Narceus spp.; pests from the class Symphyla, for example, Scutigerella immaculata; insects from the order Dermaptera, for example, Forficula auricularia; insects from the order Collembola, for example, Onychiurus spp.(e.g., Onychiurus armatus); pests from the order Isopoda, for example, Armadillidium vulgare, Oniscus asellus, Porcellio scaber; insects from the order Phthiraptera, for example, Damalinia spp., Pediculus spp. (e.g., Pediculus humanus capitis, Pediculus humanus corporis, Pediculus humanus humanus); Pthirus pubis, Haematopinus spp. (e.g., Haematopinus eurysternus, Haematopinus suis); Linognathus spp. (e.g., Linognathus vituli); Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes Examples of further pest species that may be controlled by the compounds and / or compositions identified herein include the phylum Mollusca, the class Bivalvia, e.g., Dreissena spp.; the class Gastropoda, e.g., Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea canaliclata, Succinea spp.; the class Helminthi, e.g., Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp., Dictyocaulus filaria, Diphyllobothrium latum, Dracunculus medinensis, Echinococcus granulosus, Echinococcus multilocularis, Enterobius vermicularis, Faciola spp., Haemonchus spp. (e.g., Haemonchus contortus); Heterakis spp., Hymenolepis Nanchunga, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca vo. lvulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborni, Strongyloides stercora lis, Stronyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichuria, and Wuchereria bancrofti. One method.

[14] A candidate compound selected by the method according to any one of [1] to [4].

[15] The candidate compound according to

[14] , wherein the candidate compound is a modulator that inhibits the activity of the complex.

[16] The candidate compound according to

[14] , wherein the candidate compound is a modulator that activates the complex.

[17] The candidate compound according to

[15] or

[16] , wherein the candidate compound interferes with insect behavior.

[18] The candidate compound according to

[15] or

[16] , wherein the candidate compound kills insects.

[19] The candidate compound according to

[15] or

[16] , wherein the candidate compound repels insects.

[20] A method for controlling insects, comprising applying a candidate compound selected by the method according to

[15] or

[16] to insects.

[21] The method according to

[20] , wherein the candidate compound is an inhibitor of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

[22] The method according to

[20] , wherein the candidate compound is an activator of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

[23] The method according to any one of

[20] to

[22] , wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

[24] Specific species of lepidoptera include Achroia grisella, Acleris spp. (including A. fimbriana, A. gloverana, and A. variana); Acrolepiopsis assectella 、Acronita major 、Adoxophyes spp spp.(from A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp spp. (including A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyresthia conjugella, Argyroploce spp., Argyrotaenia spp. (including A. velutiNanchunga); Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp.、Cacoecia spp.(from C.muriNanchungaC.podana);Cactoblastis cactorum、Cadra cautella、Calingo braziliensis、Caloptilis theivora、Capua reticulana、Carposina spp aridula, Cheimatobia brumata, Chilo spp. (including C. Indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp spp. 、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus);Desmia funeralis, Diaphania spp.hyalinata);Diatraea grandiosella、Diatraea saccharalis、Diphthera festiva、Earias spp.(E.insulana、E.vittella);Ecdytolopha aurantianu、Egira(=Xylomyges)curialis、Elasmopalpus lignosellus、Eldana saccharina、Endopiza viteana、Ennomos subsignaria、Eoreuma loftini、Ephestia spp thrax、Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Faronta albilinea、Feltia spp mellonella、Gracillaria spp.、Grapholite spp.(G.funebrana、G.molesta、G.inopinata); spp. (including H. armigera (=Heliothis armigera) and H. zea (=Heliothis zea)); spp.(excluding H.undalis, H. spp.rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorella, Lerodea eufala, Leucinodes orbonalis, Leucoma salicis, Leucoptera spp. (e.g., L.coffeella, L.scitella); Leuminivora lycinivorell, Lithocolletis blancardella, Lithophane antennata, Llattia octo(=Amyna axis), Lobesia botrana, Lophocampa spp., Loxagrotis albicosta, Loxostege spp. (e.g., L.sticticalis, L.cereralis); Lymantria spp. (e.g., L.dispar, L.monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta);Marasmia spp.Marmara spp.Maruca testulalis、Megalopyge lanata、Melanchra picta、Melanitis leda、Mocis spp separate、Nemapogon cloacella、Neoleucinodes elegantalis、Nepytia spp.、Nymphula spp.、Oiketicus spp.、Omiodes indicata、Omphisa anastomosalis、Operophtera brumata、Orgyia pseudotsugata、Oria spp.Orthaga thyrisalis, Ostrinia spp. (O.nubilalis); Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp licorice, Paysandisia archon, Pectinophora spp., and P. gossypiella; Peridroma saucia, Perileucoptera spp., such as P. coffeella; Phalera bucephala, Phryganidia californica, and Phthorimaea spp. (including P. operculella);Phyllocnistis citrella; Phyllonorycter spp. (including P. blancardella, P. crataegella, P. issikii, P. ringoniella); Pieris spp.(such as P. brassicae, P. rapae, P. napi);Pilocrocis tripunctata, Platypena scabra, Platynota spp.stultana); Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp. (e.g., P.sequax, P.unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana,. Sabulodes aegrotata Schizura concinna Schoenobius spp. Schreckensteinia festaliella Scirpophaga spp spp.(S.inferens、Seudyra subflava、Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.(singえば、T.ni);Night(=Scrobipalpula)absolute、Udea spp.(singhえば、U.rubigalis、U.rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, such as Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A.anxius, A.planipennis, A.sinuatus); Agriotes spp. (e.g. A.fuscicollis, A.lineatus, A.obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A.glabripennis); Anthonomus spp. (e.g. A.eugenii, A.grandis, A.pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A.linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g., B.lentis, B.pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C.assimilis, C.napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E. similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp. (e.g., M. aeneus); Melolontha spp. (e.g., M. hippocastani, M. melolontha); Metamasius hemipterus, Microtheca spp., Migdolus spp. (e.g., M. fryanus), Monochamus spp. (e.g., M. alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P.helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R. billineatus, R. ferrugineus, R. palmarum, R. phoenicis, R. vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S. granaria, S. oryzae, S. zeamais); Sphenophorus spp. (e.g., S. levis); Stegobium paniceum, Sternechus spp. (e.g., S. subsignatus); Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp.、Tenebrio molitor、Tenebrioides mauretanicus、Tribolium spp.(for example、T.castaneum); spp.quadrimaculatus, A.sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. (e.g. C.bezziana, C.hominivorax, C.macellaria); Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp. (e.g. C.hominivorax); Contarinia spp. (e.g. C.sorghicola); Cordylobia anthropophaga, Culex spp. (e.g. 、 C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g. D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g. D. suzukii), Fannia spp. (e.g. F. canicularis); Gastraphilus spp. (e.g. G. intestinalis); Geomyza tipunctata, Glossina spp. (e.g. G. fuscipes, G. morsitans, G. palpalis, G. tachinoides); Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp. (e.g. H. platura); Hypoderma spp.(eg H.lineata);Hyppobosca spp.、Hydrellia philippina、Leptoconops torrens、Liriomyza spp.(eg L.sativae、L.trifolii);Lucilia spp.(eg L.caprina、L.cuprina、L.sericata);Lycoria pectoralis、Mansonia titillanus、Mayetiola spp.(eg M.destructor); spp. (for example, M.autumnalis, M. domestica); Muscina stablens, Oestrus spp. (for example, O. ovis);frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp.; Insects from the order Thysanoptera, e.g., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp. (e.g., F.fusca, F.occidentalis, F.tritici); Heliothrips spp., Hercinothrips femoralis, Kakothrips spp., Microcephalothrips abdominalis, Neohydatothrips samayunkur, Pezothrips kellyanus, Rhipiphorothrips cruentatus, Scirtothrips spp. (e.g., S.citri, S.dorsalis, S.perseae);Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequence、Thrips spp. (including T.imagines, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, and T.tabaci); spp.(such as A.hilare);Acyrthosipon spp.(such as A.onobrychis)A.pisum);Adelges laricis, Adelges tsugae, Adelphocoris spp.(such as A.rapidus,A.superbus);Aeneolamia spp. Agonoscena spp. Aulacorthum solani Aleurocanthus woglumi Aleurodes spp. Aleurodicus disperses Aleurolobus barodensis Aleurothrixus spp. Amrasca spp spp.、Anuraphis cardui、Aonidiella spp.、Aphanostigma pyri、Aphidula nasturtii、Aphis spp.(including A. craccivora, A. fabae, A. forbesi, A. gossypii, A. grossulariae, A. maidiradicis, A. pomi, A. sambuci, A. schneideri, A. spiraecola); Arboridia apicalis, Arilus critatus, Aspidiella spp., Aspidiotus spp., Atanus spp., Aulacaspis yasumatsui, Aulacorthum solani, Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp tabaci));Blissus spp.leucopterus; Brachycaudus spp. (e.g., B. cardui, B. helichrysi, B. persicae, B. prunicola); Brachycolus spp., Brachycorynella asparagi, Brevicoryne brassicae, Cacopsylla spp. (e.g., C. fulguralis, C. pyricola (Psylla piri); Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes ceriferus, Cerosipha gossypii, Chaetosiphon fragaefolii, Chionaspis tegalensis, Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila, Cimex spp. (e.g., C. hemipterus, C. lectularius); Coccomytilus halli, Coccus spp. (e.g., C. hesperidum, C. pseudomagnoliarum); Corythucha arcuata, Creontiades dilutus, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasynus piperis, Dialeurodes spp. (e.g., D. citrifolii); Dalbulus maidis, Diaphorina spp. (e.g., D. citri); Diaspis spp. (e.g., D. bromeliae); Dichelops furcatus, Diconocoris hewetti, Doralis spp.、Dreyfusia nordmannianae、Dreyfusia piceae、Drosicha spp.、Dysaphis spp.(eg D.plantaginea、D.pyri、D.radicola);Dysaulacorthum pseudosolani、Dysdercus spp.(eg D.cingulatus、D.intermedius);Dysmicoccus spp.、Edessa spp、Geocoris spp、Empoasca spp.(eg E.fabae、E.solana);Epidiaspis leperii、Eriosoma spp.(eg E.lanigerum、E.pyricola);Erythroneura spp.、Eurygaster spp.(eg E.integriceps);Euscelis bilobatus、Euschistus spp.(eg E.heros、E.impictiventris、E.servus);Fiorinia theae、Geococcus coffeae、Glycaspis brimblecombei、Halyomorpha spp.(eg H.halys);Heliopeltis spp.、Homalodisca vitripennis (=H.coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp. (e.g. I.purchase); Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp. (e.g. L.ulmi); Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp. (e.g. L.hesperus, L.lineolaris, L.pratensis);. Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp. (e.g. M.rosae, M.avenae, M.euphorbiae); Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (=Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis coryli, Murgantia spp., Myzus spp. (e.g. M.ascalonicus, M.cerasi, M.nicotianae, M.persicae, M.varians); Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp. (e.g. N.malayanus, N.nigropictus, N.parvus, N.virescens); Nezara spp.(eg N.viridula);Nilaparvata lugens、Nysius huttoni、Oebalus spp.(eg O.pugnax);Oncometopia spp.、Orthezia praelonga、Oxycaraenus hyalinipennis、Parabemisia myricae、Parlatoria spp.、Parthenolecanium spp.(eg、P.corni、P.persicae);Pemphigus spp.(eg、P.bursarius、P.populivenae);Peregrinus maidis、Perkinsiella saccharicida、Phenacoccus spp. (for example、P. aceris、P.gossypii); Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp. (e.g. P.devastatrix), Piesma quadrata, Piezodorus spp. (e.g. P.guildinii); Pinnaspis aspidistrae, Planococcus spp. (e.g. P.citri, P.ficus); Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudacysta persea, Pseudaulacaspis pentagona, Pseudoococcus spp. (e.g. P.comstocki); Psylla spp. (e.g. P.mali); Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp. (e.g. Q.perniciosus); Quesada gigas, Rastroccus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp. (e.g., R. pseudobrassicas, R. insertum, R. maidis, R. padi); Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulatus, Sitobion avenae, Sogata spp.、Sogatella furcifera、Solubea insularis、Spissistilus festinus(=Stictocephala festina); spp.(including T.accerra;T.perditor);Tibraca spp., Tomaspis spp., Toxoptera spp.(including T.aurantii);Trialeurodes spp.(including T.abutilonea, T.ricini, and T.vaporariorum);Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp.(including U.citri,U.yanonensis);Viteus vitifolii;The list of Hymenoptera includes Acanthomyops interjectus, Athalia rosae, Atta spp (including A. capiguara, A. cephalotes, A. cephalotes, A. laevigata, A. robusta, A. sexdens, A. texana), Bombus spp., Brachymyrmex spp., and Camponotus spp (from C. floridanus, C. pennsylvanicus, C. modoc); Cardiocondyla nuda, Chalibion ​​sp, Crematogaster spp, Dasymutilla occidentalis, Diprion spp, Dolichovespula maculata, Dorymyrmex spp, Dryocosmus kuriphilus, Formica spp, Hoplocampa spp., (excluding H.minuta, H. testudinea); Iridomyrmex humilis, Lasius spp.niger) Linepithema humile Liometopum spp Leptocybe invasa Monomorium spp(M.pharaonis) Monomorium Nylandria fulva Pachycondyla chinensis Paratrechina longicornis Paravespula spp(such as P. germanica, P. pennsylvanica, P. vulgaris);Pheidole spp(such as P. megacephala);Pogonomyrmex spp(such as P. barbatus, P. californicus), Polistes rubiginosa、Prenolepis impairs、Pseudomyrmex gracilis、Schelipron spp、Sirex cyaneus、Solenopsis spp(S.geminata、S.invicta、S.molesta、S.richteri、S.xyloni) Sphecius speciosus spp (including V.squamosal);Wasmannia auropunctata, Xylocopa sp.;Orthoptera species include Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, and Ceuthophilus spp. Diastrammena asynamora. Dociostaurus maroccanus spp.(such as L. migratoria, L. pardalina);Melanoplus spp.(such as M. spp.).bivittatus, M. femurrubrum, M. mexicanus, M. sanguinipes, M. spretus); spp(S.americana, S.gregaria), Stemopelmatus spp, Tachycines asynamorus, and Zonozerus variegatus;Arachnidae, Argasidae, Ixodidae, Sarcoptidae (Acari), Amblyomma spp.(A.americanum, A.variegatum, A.maculatum), Argas spp., (A. persicu), Boophilus spp., B. annulatus, B. decoloratus, B. microplus) Dermacentor spp (including D. silvarum, D. andersoni, D. variabilis), Hyalomma spp. (including H. truncatum), Ixodes spp.(I.ricinus, I.rubicundus, I.scapularis, I.holocyclus, I.pacificus) Rhipicephalus sanguineus Ornithodorus spp.(excluding O. moubata, O. hermsi, O. turicata), Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp., (excluding P.ovis), Rhipicephalus spp(including R.sanguineus, R.appendiculatus), Rhipicephalus evertsi), Rhizoglyphus spp; Sarcoptes spp. (including S. Scabiei); and Aceria spp (including A. sheldoni, A. anthocoptes), Acallitus spp; Aculops spp.pelekassi); Aculus spp. (e.g., A. schlechtendali); Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; the Eriophyidae family, including Eriophytes ribis and Eriophyes spp. (e.g., Eriophyes sheldoni); the Tarsonemiidae family, including Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp., and Stenotarsonemus spinki; the Tenuipalpidae family, including Brevipalpus spp. (e.g., B. phoenicis); Eotetranychus spp., Eut. The Tetranychidae family includes species such as etranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp. (e.g., T. cinnabarinus, T. evansi, T. kanzawai, T. pacificus, T. phaseulus, T. telarius, and T. urticae); Bryobia praetiosa; Panonychus spp. (e.g., P. ulmi, P. citri); Metatetranychus spp., and Oligonychus spp. (e.g., O. pratensis, O. perseae); Vasates lycopersici; Raoiella indica; Carpoglyphidae, including Carpoglyphus spp.; Penthaleidae spp. (e.g., Halotydeus destructor); Demodicidae, including species such as Demodex spp.; Trombicula family Trombicidea, including spp.; family Macronyssidae, including Ornothonyssus spp.; family Pyemotidae, including Pyemotes tritici; Tyrophagus putrescentiae; family Acaridae, including Acarus siro; family Araneida, including Latrodectus mactans, Tegenaria agrestis, Chiracanthium spp., Lycosa spp., Achaearanea tepidariorum, and Loxosceles reclusa; pests from the phylum Nematoda, for example, plant-parasitic nematodes (e.g., root-knot nematodes) Meloidogyne spp. (e.g., M. hapla, M. incognita, M. javanica); cyst nematodes Globodera spp. (e.g., G. rostochiensis); Heterodera (e.g. H. avenae, H. glycines, H. schachtii, H. trifolii); Seed gall nematodes Anguina spp.Stem and foliar nematodes Aphelenchoides spp. (e.g., A. besseyi); Sting nematodes Belonolaimus spp. (e.g., B. longicaudatus); Pine nematodes Bursaphelenchus spp. (e.g., B. lignicolus, B. xylophilus); Ring nematodes Criconema spp.; Criconemella spp. (e.g., C. xenoplax and C. ornata); and Criconemoides spp. (e.g., Criconemoides informis; Mesocriconema spp.); Stem and bulb nematodes Ditylenchus spp. (e.g., D. destructor, D. dipsaci); Awl nematodes (Dolichodorus spp.); spiral nematodes (Heliocotylenchus multicinctus); sheath and sheathoid nematodes (Hemicycliophora spp. and Hemicriconemoides spp.); Hirshmanniella spp.; lance nematodes (Hoploaimus spp.); false rootknot nematodes (Nacobbus spp.); needle nematodes (Longidorus spp. (e.g., L. elongatus); root-lesion nematodes (Pratylenchus spp. (e.g., P. brachyurus, P. neglectus, P. penetrans, P. curvitatus, P. goodeyi); Burrowing nematodes Radopholus spp. (e.g., R. similis; Rhadopholus spp.; Rhodopholus spp.;Reniform nematodes Rotylenchus spp. (e.g., R. robustus, R. reniformis); Scutellonema spp.;Stubby-root nematodes Trichodorus spp. (e.g., T. obtusus, T. primitivus); Paratrichodorus spp. (e.g., P. minor);Stunt nematodes Tylenchorhynchus spp. (e.g., T. claytoni, T. dubius);Citrus nematodes Tylenchulus spp. (e.g., T. semipenetrans);Dagger nematodes Xiphinema spp.;and other plant-parasitic nematode species;Insecta from the order Isoptera, for example, Calotermes flavicollis, Coptotermes spp (e.g. C. formosanus, C. gestroi, C. acinaciformis); Cornitermes cumulans, Cryptotermes spp (e.g. C. brevis, C. cavifrons); Globitermes sulfureus, Heterotermes spp (e.g. H. aureus, H. longiceps, H. tenuis); Leucotermes flavipes, Odontotermes spp., Incisitermes spp (e.g. I. minor, I. Snyder); Marginitermes hubbardi, Mastotermes spp (e.g. M. darwiniensis), Neocapritermes spp (e.g. N.opacus, N.parvus); Neotermes spp, Procornitermes spp, Zootermopsis spp. (e.g. Z. angusticollis, Z. nevadensis), Reticulitermes spp. (e.g. R. hesperus, R. tibialis, R. speratus, R. flavipes, R. grassei, R. lucifugus, R.santonensis, R.virginicus);Termes natalensis;Insects from the order Blattaria, such as Blatta spp (e.g. B.orientalis, B.lateralis);Blattella spp (e.g. B.asahinae, B.germanica);Leucophaea maderae, Panchlora nivea, Periplaneta spp (e.g. P.americana, P.australasiae, P.brunnea, P.fuligginosa, P.japonica); Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnoscelus surinamensis; insects from the order Siphonoptera, e.g. Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp (e.g. C. felis, C. canis), Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus; insects from the order Thysanura, for example, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica; pests from the class Chilopoda, for example, Geophilus spp., Scutigera spp. (e.g., Scutigera coleoptrata); pests from the class Diplopoda, for example, Blaniulus guttulatus, Julus spp., Narceus spp.; pests from the class Symphyla, for example, Scutigerella immaculata; insects from the order Dermaptera, for example, Forficula auricularia; insects from the order Collembola, for example, Onychiurus spp.(e.g., Onychiurus armatus); pests from the order Isopoda, for example, Armadillidium vulgare, Oniscus asellus, Porcellio scaber; insects from the order Phthiraptera, for example, Damalinia spp., Pediculus spp. (e.g., Pediculus humanus capitis, Pediculus humanus corporis, Pediculus humanus humanus); Pthirus pubis, Haematopinus spp. (e.g., Haematopinus eurysternus, Haematopinus suis); Linognathus spp. (e.g., Linognathus vituli); Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes Examples of further pest species that may be controlled by the compounds and / or compositions identified herein include the phylum Mollusca, the class Bivalvia, e.g., Dreissena spp.; the class Gastropoda, e.g., Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea canaliclata, Succinea spp.; the class Helminthi, e.g., Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp., Dictyocaulus filaria, Diphyllobothrium latum, Dracunculus medinensis, Echinococcus granulosus, Echinococcus multilocularis, Enterobius vermicularis, Faciola spp., Haemonchus spp. (e.g., Haemonchus contortus); Heterakis spp., Hymenolepis Nanchunga, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca vo. lv ulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborni, Strongyloides stercora lis, Stronyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichuria, and Wuchereria bancrofti.

[25] (a) a first expression vector comprising a first nucleic acid molecule encoding a Nanchung protein; (b) a second expression vector comprising a second nucleic acid molecule encoding a Water witch protein.

[26]

[25] The combination of expression vectors described in

[25] , wherein an insect TRP channel is formed upon simultaneous expression of the first nucleic acid molecule and the second nucleic acid molecule in a cell containing both the first expression vector and the second expression vector.

[27] The combination of expression vectors according to

[25] or

[26] , wherein the first nucleic acid molecule comprises a nucleic acid sequence of SEQ ID NO: 1 to 27.

[28] The combination of expression vectors according to

[25] or

[26] , wherein the second nucleic acid molecule comprises a nucleic acid sequence of SEQ ID NO: 28 to 63.

[29] (a) a first nucleic acid molecule encoding a Nanchung protein; (b) a second nucleic acid molecule encoding a Water witch protein.

[30]

[29] The expression vector according to

[29] , wherein an insect TRP channel is formed upon co-expression in a cell containing both the first nucleic acid molecule and the second nucleic acid molecule.

[31] The expression vector according to

[29] or

[30] , wherein the first nucleic acid molecule comprises a nucleic acid sequence of SEQ ID NO: 1 to 27.

[32] The expression vector according to

[29] or

[30] , wherein the second nucleic acid molecule comprises a nucleic acid sequence of SEQ ID NO: 28 to 63.

[33] A cell comprising the expression vector according to

[29] or

[30] .

[34] The cell according to

[33] , wherein the Nanchung protein and the Water witch protein are co-expressed in the cell.

[35] The cells described in

[34] , including any cells that do not normally co-express Nanchung and Water witch proteins.

[36] The cell according to

[35] , wherein the cell is an insect cell, an amphibian cell, or a mammalian cell.

[37] 1. A method for determining whether a candidate compound directly binds to a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein, comprising: (a) providing a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein; (b) contacting the complex with a candidate compound; (c) determining the binding affinity of the candidate compound to the complex.

[38] The method according to

[37] , wherein the candidate compound is an unlabeled compound.

[39] The method according to

[37] , wherein the candidate compound is a labeled compound.

[40] The binding affinity is K d and / or K i The method according to any one of

[37] to

[39] , wherein the equilibrium binding constant is determined using one or more equilibrium binding constants including:

[41] The equilibrium dissociation constant (K d ) is about 3 × 10 -7 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[42] The equilibrium dissociation constant (K d ) is about 3 × 10 -8 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[43] The equilibrium dissociation constant (K d ) is about 3 × 10 -9 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[44] The equilibrium dissociation constant (K d ) is about 3 × 10 -10 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[45] The equilibrium dissociation constant (K d ) is about 3 × 10 -11 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[46] The equilibrium dissociation constant (K d ) is about 3 × 10 -12 The method according to any one of

[37] to

[39] , wherein M is equal to or less than M.

[47] The method according to

[39] , wherein the candidate compound is labeled with a radioactive isotope.

[48] The method according to

[47] , wherein the candidate compound is labeled with a fluorophore.

[49] The method according to

[47] or

[48] , wherein the labeled compound is afidopiropene.

[50] 1. A method for classifying candidate compounds, comprising: (a) a binding assay step of contacting a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein with the candidate compound and determining the binding affinity of the candidate compound to the complex; (b) identifying a mechanism of action class of the candidate compound.

[51] The bonding step comprises: (1) providing the complex; (2) contacting the complex with a labeled or unlabeled candidate compound; (3) measuring the amount of the candidate compound bound to the complex; (4) the equilibrium dissociation constant (K d ), wherein the Kd However, below, (I)(a) Maximum connection (B max ) and (b) the concentration of the candidate compound required to achieve 50% of maximum binding, (II) the association of the candidate compound (K on ) and dissociation (K off ) rate measurement (where K d =K off / K on (III) fluorescence polarization assay, (IV) surface plasmon resonance, (V) isothermal calorimetry, or (VI) thermophoresis; (5) comparing the equilibrium dissociation constant of the candidate compound with the equilibrium dissociation constants of other candidate compounds; or (6) The equilibrium dissociation constant of the candidate compound is compared with the equilibrium inhibition constant (K i ) comparing the signal with the signal.

[52] The bonding step comprises: (1) providing the complex; (2) contacting the complex with the candidate compound and one labeled compound that has a high binding affinity for the complex; (3) increasing the concentration of unlabeled candidate compound available for contact with the complex until binding of labeled compound to the complex approaches 0%; (4) determining the concentration of the candidate compound required to reduce the amount of bound labeled compound by 50%; (5) The equilibrium inhibition constant (K i ), (6) The equilibrium inhibition constant (K i ) with the inhibition constants of other candidate compounds; (7) The equilibrium inhibition constant of the candidate compound is calculated based on the equilibrium dissociation constant (K d ), or (8) identifying one or more candidate compounds having high affinity for the complex.

[53] The method of

[37] or

[50] , wherein binding of the candidate compound to the complex interferes with insect behavior.

[54] The method of

[37] or

[50] , wherein binding of the candidate compound to the complex kills the insect.

[55] The method of

[37] or

[50] , wherein binding of the candidate compound to the complex repels insects.

[56] The method according to

[37] or

[50] , wherein the candidate compound is selected from the group consisting of small organic molecules, small inorganic molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

[57] A composition comprising one or more candidate compounds that directly bind to a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

[58] A composition comprising one or more candidate compounds selected by the method according to

[37] or

[50] .

[59]

[57] A method of insect control comprising applying the composition of

[57] to one or more of insects, insect habitats, bait, traps, and / or insect feeding sources.

[60] The method according to

[59] , wherein the candidate compound is an inhibitor of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

[61] The method according to

[59] , wherein the candidate compound is an activator of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

[62] The method according to

[37] ,

[50] ,

[59] ,

[60] or

[61] , wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

[63] Specific species of lepidoptera include Achroia grisella, Acleris spp. (including A. fimbriana, A. gloverana, and A. variana); Acrolepiopsis assectella 、Acronita major 、Adoxophyes spp spp.(from A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp spp. (including A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyresthia conjugella, Argyroploce spp., Argyrotaenia spp. (including A. velutiNanchunga); Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp.、Cacoecia spp.(from C.muriNanchungaC.podana);Cactoblastis cactorum、Cadra cautella、Calingo braziliensis、Caloptilis theivora、Capua reticulana、Carposina spp aridula, Cheimatobia brumata, Chilo spp. (including C. Indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp spp. 、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus);Desmia funeralis, Diaphania spp.hyalinata);Diatraea grandiosella、Diatraea saccharalis、Diphthera festiva、Earias spp.(E.insulana、E.vittella);Ecdytolopha aurantianu、Egira(=Xylomyges)curialis、Elasmopalpus lignosellus、Eldana saccharina、Endopiza viteana、Ennomos subsignaria、Eoreuma loftini、Ephestia spp thrax、Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Faronta albilinea、Feltia spp mellonella、Gracillaria spp.、Grapholite spp.(G.funebrana、G.molesta、G.inopinata); spp. (including H. armigera (=Heliothis armigera) and H. zea (=Heliothis zea)); spp.(excluding H.undalis, H. spp.rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorella, Lerodea eufala, Leucinodes orbonalis, Leucoma salicis, Leucoptera spp. (e.g., L.coffeella, L.scitella); Leuminivora lycinivorell, Lithocolletis blancardella, Lithophane antennata, Llattia octo(=Amyna axis), Lobesia botrana, Lophocampa spp., Loxagrotis albicosta, Loxostege spp. (e.g., L.sticticalis, L.cereralis); Lymantria spp. (e.g., L.dispar, L.monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta);Marasmia spp.Marmara spp.Maruca testulalis、Megalopyge lanata、Melanchra picta、Melanitis leda、Mocis spp separate、Nemapogon cloacella、Neoleucinodes elegantalis、Nepytia spp.、Nymphula spp.、Oiketicus spp.、Omiodes indicata、Omphisa anastomosalis、Operophtera brumata、Orgyia pseudotsugata、Oria spp.Orthaga thyrisalis, Ostrinia spp. (O.nubilalis); Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp licorice, Paysandisia archon, Pectinophora spp., and P. gossypiella; Peridroma saucia, Perileucoptera spp., such as P. coffeella; Phalera bucephala, Phryganidia californica, and Phthorimaea spp. (including P. operculella);Phyllocnistis citrella; Phyllonorycter spp. (including P. blancardella, P. crataegella, P. issikii, P. ringoniella); Pieris spp.(such as P. brassicae, P. rapae, P. napi);Pilocrocis tripunctata, Platypena scabra, Platynota spp.stultana); Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp. (e.g., P.sequax, P.unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana,. Sabulodes aegrotata Schizura concinna Schoenobius spp. Schreckensteinia festaliella Scirpophaga spp spp.(S.inferens、Seudyra subflava、Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.(singえば、T.ni);Night(=Scrobipalpula)absolute、Udea spp.(singhえば、U.rubigalis、U.rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, such as Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A.anxius, A.planipennis, A.sinuatus); Agriotes spp. (e.g. A.fuscicollis, A.lineatus, A.obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A.glabripennis); Anthonomus spp. (e.g. A.eugenii, A.grandis, A.pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A.linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g., B.lentis, B.pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C.assimilis, C.napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E. similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp. (e.g., M. aeneus); Melolontha spp. (e.g., M. hippocastani, M. melolontha); Metamasius hemipterus, Microtheca spp., Migdolus spp. (e.g., M. fryanus), Monochamus spp. (e.g., M. alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P.helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R. billineatus, R. ferrugineus, R. palmarum, R. phoenicis, R. vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S. granaria, S. oryzae, S. zeamais); Sphenophorus spp. (e.g., S. levis); Stegobium paniceum, Sternechus spp. (e.g., S. subsignatus); Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp.、Tenebrio molitor、Tenebrioides mauretanicus、Tribolium spp.(for example、T.castaneum); spp.quadrimaculatus, A.sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. (e.g. C.bezziana, C.hominivorax, C.macellaria); Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp. (e.g. C.hominivorax); Contarinia spp. (e.g. C.sorghicola); Cordylobia anthropophaga, Culex spp. (e.g. 、 C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g. D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g. D. suzukii), Fannia spp. (e.g. F. canicularis); Gastraphilus spp. (e.g. G. intestinalis); Geomyza tipunctata, Glossina spp. (e.g. G. fuscipes, G. morsitans, G. palpalis, G. tachinoides); Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp. (e.g. H. platura); Hypoderma spp.(eg H.lineata);Hyppobosca spp.、Hydrellia philippina、Leptoconops torrens、Liriomyza spp.(eg L.sativae、L.trifolii);Lucilia spp.(eg L.caprina、L.cuprina、L.sericata);Lycoria pectoralis、Mansonia titillanus、Mayetiola spp.(eg M.destructor); spp. (for example, M.autumnalis, M. domestica); Muscina stablens, Oestrus spp. (for example, O. ovis);frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp.; Insects from the order Thysanoptera, e.g., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp. (e.g., F.fusca, F.occidentalis, F.tritici); Heliothrips spp., Hercinothrips femoralis, Kakothrips spp., Microcephalothrips abdominalis, Neohydatothrips samayunkur, Pezothrips kellyanus, Rhipiphorothrips cruentatus, Scirtothrips spp. (e.g., S.citri, S.dorsalis, S.perseae);Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequence、Thrips spp. (including T.imagines, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, and T.tabaci); spp.(such as A.hilare);Acyrthosipon spp.(such as A.onobrychis)A.pisum);Adelges laricis, Adelges tsugae, Adelphocoris spp.(such as A.rapidus,A.superbus);Aeneolamia spp. Agonoscena spp. Aulacorthum solani Aleurocanthus woglumi Aleurodes spp. Aleurodicus disperses Aleurolobus barodensis Aleurothrixus spp. Amrasca spp spp.、Anuraphis cardui、Aonidiella spp.、Aphanostigma pyri、Aphidula nasturtii、Aphis spp.(including A. craccivora, A. fabae, A. forbesi, A. gossypii, A. grossulariae, A. maidiradicis, A. pomi, A. sambuci, A. schneideri, A. spiraecola); Arboridia apicalis, Arilus critatus, Aspidiella spp., Aspidiotus spp., Atanus spp., Aulacaspis yasumatsui, Aulacorthum solani, Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp tabaci));Blissus spp.leucopterus; Brachycaudus spp. (e.g., B. cardui, B. helichrysi, B. persicae, B. prunicola); Brachycolus spp., Brachycorynella asparagi, Brevicoryne brassicae, Cacopsylla spp. (e.g., C. fulguralis, C. pyricola (Psylla piri); Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes ceriferus, Cerosipha gossypii, Chaetosiphon fragaefolii, Chionaspis tegalensis, Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila, Cimex spp. (e.g., C. hemipterus, C. lectularius); Coccomytilus halli, Coccus spp. (e.g., C. hesperidum, C. pseudomagnoliarum); Corythucha arcuata, Creontiades dilutus, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasynus piperis, Dialeurodes spp. (e.g., D. citrifolii); Dalbulus maidis, Diaphorina spp. (e.g., D. citri); Diaspis spp. (e.g., D. bromeliae); Dichelops furcatus, Diconocoris hewetti, Doralis spp.、Dreyfusia nordmannianae、Dreyfusia piceae、Drosicha spp.、Dysaphis spp.(eg D.plantaginea、D.pyri、D.radicola);Dysaulacorthum pseudosolani、Dysdercus spp.(eg D.cingulatus、D.intermedius);Dysmicoccus spp.、Edessa spp、Geocoris spp、Empoasca spp.(eg E.fabae、E.solana);Epidiaspis leperii、Eriosoma spp.(eg E.lanigerum、E.pyricola);Erythroneura spp.、Eurygaster spp.(eg E.integriceps);Euscelis bilobatus、Euschistus spp.(eg E.heros、E.impictiventris、E.servus);Fiorinia theae、Geococcus coffeae、Glycaspis brimblecombei、Halyomorpha spp.(eg H.halys);Heliopeltis spp.、Homalodisca vitripennis (=H.coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp. (e.g. I.purchase); Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp. (e.g. L.ulmi); Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp. (e.g. L.hesperus, L.lineolaris, L.pratensis);. Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp. (e.g. M.rosae, M.avenae, M.euphorbiae); Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (=Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis coryli, Murgantia spp., Myzus spp. (e.g. M.ascalonicus, M.cerasi, M.nicotianae, M.persicae, M.varians); Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp. (e.g. N.malayanus, N.nigropictus, N.parvus, N.virescens); Nezara spp.(eg N.viridula);Nilaparvata lugens、Nysius huttoni、Oebalus spp.(eg O.pugnax);Oncometopia spp.、Orthezia praelonga、Oxycaraenus hyalinipennis、Parabemisia myricae、Parlatoria spp.、Parthenolecanium spp.(eg、P.corni、P.persicae);Pemphigus spp.(eg、P.bursarius、P.populivenae);Peregrinus maidis、Perkinsiella saccharicida、Phenacoccus spp. (for example、P. aceris、P.gossypii); Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp. (e.g. P.devastatrix), Piesma quadrata, Piezodorus spp. (e.g. P.guildinii); Pinnaspis aspidistrae, Planococcus spp. (e.g. P.citri, P.ficus); Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudacysta persea, Pseudaulacaspis pentagona, Pseudoococcus spp. (e.g. P.comstocki); Psylla spp. (e.g. P.mali); Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp. (e.g. Q.perniciosus); Quesada gigas, Rastroccus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp. (e.g., R. pseudobrassicas, R. insertum, R. maidis, R. padi); Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulatus, Sitobion avenae, Sogata spp.、Sogatella furcifera、Solubea insularis、Spissistilus festinus(=Stictocephala festina); spp.(including T.accerra;T.perditor);Tibraca spp., Tomaspis spp., Toxoptera spp.(including T.aurantii);Trialeurodes spp.(including T.abutilonea, T.ricini, and T.vaporariorum);Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp.(including U.citri,U.yanonensis);Viteus vitifolii;The list of Hymenoptera includes Acanthomyops interjectus, Athalia rosae, Atta spp (including A. capiguara, A. cephalotes, A. cephalotes, A. laevigata, A. robusta, A. sexdens, A. texana), Bombus spp., Brachymyrmex spp., and Camponotus spp (from C. floridanus, C. pennsylvanicus, C. modoc); Cardiocondyla nuda, Chalibion ​​sp, Crematogaster spp, Dasymutilla occidentalis, Diprion spp, Dolichovespula maculata, Dorymyrmex spp, Dryocosmus kuriphilus, Formica spp, Hoplocampa spp., (excluding H.minuta, H. testudinea); Iridomyrmex humilis, Lasius spp.niger) Linepithema humile Liometopum spp Leptocybe invasa Monomorium spp(M.pharaonis) Monomorium Nylandria fulva Pachycondyla chinensis Paratrechina longicornis Paravespula spp(such as P. germanica, P. pennsylvanica, P. vulgaris);Pheidole spp(such as P. megacephala);Pogonomyrmex spp(such as P. barbatus, P. californicus), Polistes rubiginosa、Prenolepis impairs、Pseudomyrmex gracilis、Schelipron spp、Sirex cyaneus、Solenopsis spp(S.geminata、S.invicta、S.molesta、S.richteri、S.xyloni) Sphecius speciosus spp (including V.squamosal);Wasmannia auropunctata, Xylocopa sp.;Orthoptera species include Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, and Ceuthophilus spp. Diastrammena asynamora. Dociostaurus maroccanus spp.(such as L. migratoria, L. pardalina);Melanoplus spp.(such as M. spp.).bivittatus, M. femurrubrum, M. mexicanus, M. sanguinipes, M. spretus); spp(S.americana, S.gregaria), Stemopelmatus spp, Tachycines asynamorus, and Zonozerus variegatus;Arachnidae, Argasidae, Ixodidae, Sarcoptidae (Acari), Amblyomma spp.(A.americanum, A.variegatum, A.maculatum), Argas spp., (A. persicu), Boophilus spp., B. annulatus, B. decoloratus, B. microplus) Dermacentor spp (including D. silvarum, D. andersoni, D. variabilis), Hyalomma spp. (including H. truncatum), Ixodes spp.(I.ricinus, I.rubicundus, I.scapularis, I.holocyclus, I.pacificus) Rhipicephalus sanguineus Ornithodorus spp.(excluding O. moubata, O. hermsi, O. turicata), Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp., (excluding P.ovis), Rhipicephalus spp(including R.sanguineus, R.appendiculatus), Rhipicephalus evertsi), Rhizoglyphus spp; Sarcoptes spp. (including S. Scabiei); and Aceria spp (including A. sheldoni, A. anthocoptes), Acallitus spp; Aculops spp.pelekassi); Aculus spp. (e.g., A. schlechtendali); Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; the Eriophyidae family, including Eriophytes ribis and Eriophyes spp. (e.g., Eriophyes sheldoni); the Tarsonemiidae family, including Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp., and Stenotarsonemus spinki; the Tenuipalpidae family, including Brevipalpus spp. (e.g., B. phoenicis); Eotetranychus spp., Eut. The Tetranychidae family includes species such as etranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp. (e.g., T. cinnabarinus, T. evansi, T. kanzawai, T. pacificus, T. phaseulus, T. telarius, and T. urticae); Bryobia praetiosa; Panonychus spp. (e.g., P. ulmi, P. citri); Metatetranychus spp., and Oligonychus spp. (e.g., O. pratensis, O. perseae); Vasates lycopersici; Raoiella indica; Carpoglyphidae, including Carpoglyphus spp.; Penthaleidae spp. (e.g., Halotydeus destructor); Demodicidae, including species such as Demodex spp.; Trombicula family Trombicidea, including spp.; family Macronyssidae, including Ornothonyssus spp.; family Pyemotidae, including Pyemotes tritici; Tyrophagus putrescentiae; family Acaridae, including Acarus siro; family Araneida, including Latrodectus mactans, Tegenaria agrestis, Chiracanthium spp., Lycosa spp., Achaearanea tepidariorum, and Loxosceles reclusa; pests from the phylum Nematoda, for example, plant-parasitic nematodes (e.g., root-knot nematodes) Meloidogyne spp. (e.g., M. hapla, M. incognita, M. javanica); cyst nematodes Globodera spp. (e.g., G. rostochiensis); Heterodera (e.g. H. avenae, H. glycines, H. schachtii, H. trifolii); Seed gall nematodes Anguina spp.Stem and foliar nematodes Aphelenchoides spp. (e.g., A. besseyi); Sting nematodes Belonolaimus spp. (e.g., B. longicaudatus); Pine nematodes Bursaphelenchus spp. (e.g., B. lignicolus, B. xylophilus); Ring nematodes Criconema spp.; Criconemella spp. (e.g., C. xenoplax and C. ornata); and Criconemoides spp. (e.g., Criconemoides informis; Mesocriconema spp.); Stem and bulb nematodes Ditylenchus spp. (e.g., D. destructor, D. dipsaci); Awl nematodes (Dolichodorus spp.); spiral nematodes (Heliocotylenchus multicinctus); sheath and sheathoid nematodes (Hemicycliophora spp. and Hemicriconemoides spp.); Hirshmanniella spp.; lance nematodes (Hoploaimus spp.); false rootknot nematodes (Nacobbus spp.); needle nematodes (Longidorus spp. (e.g., L. elongatus); root-lesion nematodes (Pratylenchus spp. (e.g., P. brachyurus, P. neglectus, P. penetrans, P. curvitatus, P. goodeyi); Burrowing nematodes Radopholus spp. (e.g., R. similis; Rhadopholus spp.; Rhodopholus spp.;Reniform nematodes Rotylenchus spp. (e.g., R. robustus, R. reniformis); Scutellonema spp.;Stubby-root nematodes Trichodorus spp. (e.g., T. obtusus, T. primitivus); Paratrichodorus spp. (e.g., P. minor);Stunt nematodes Tylenchorhynchus spp. (e.g., T. claytoni, T. dubius);Citrus nematodes Tylenchulus spp. (e.g., T. semipenetrans);Dagger nematodes Xiphinema spp.;and other plant-parasitic nematode species;Insecta from the order Isoptera, for example, Calotermes flavicollis, Coptotermes spp (e.g. C. formosanus, C. gestroi, C. acinaciformis); Cornitermes cumulans, Cryptotermes spp (e.g. C. brevis, C. cavifrons); Globitermes sulfureus, Heterotermes spp (e.g. H. aureus, H. longiceps, H. tenuis); Leucotermes flavipes, Odontotermes spp., Incisitermes spp (e.g. I. minor, I. Snyder); Marginitermes hubbardi, Mastotermes spp (e.g. M. darwiniensis), Neocapritermes spp (e.g. N.opacus, N.parvus); Neotermes spp, Procornitermes spp, Zootermopsis spp. (e.g. Z. angusticollis, Z. nevadensis), Reticulitermes spp. (e.g. R. hesperus, R. tibialis, R. speratus, R. flavipes, R. grassei, R. lucifugus, R.santonensis, R.virginicus);Termes natalensis;Insects from the order Blattaria, such as Blatta spp (e.g. B.orientalis, B.lateralis);Blattella spp (e.g. B.asahinae, B.germanica);Leucophaea maderae, Panchlora nivea, Periplaneta spp (e.g. P.americana, P.australasiae, P.brunnea, P.fuligginosa, P.japonica); Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnoscelus surinamensis; insects from the order Siphonoptera, e.g. Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp (e.g. C. felis, C. canis), Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus; insects from the order Thysanura, for example, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica; pests from the class Chilopoda, for example, Geophilus spp., Scutigera spp. (e.g., Scutigera coleoptrata); pests from the class Diplopoda, for example, Blaniulus guttulatus, Julus spp., Narceus spp.; pests from the class Symphyla, for example, Scutigerella immaculata; insects from the order Dermaptera, for example, Forficula auricularia; insects from the order Collembola, for example, Onychiurus spp.(e.g., Onychiurus armatus); pests from the order Isopoda, for example, Armadillidium vulgare, Oniscus asellus, Porcellio scaber; insects from the order Phthiraptera, for example, Damalinia spp., Pediculus spp. (e.g., Pediculus humanus capitis, Pediculus humanus corporis, Pediculus humanus humanus); Pthirus pubis, Haematopinus spp. (e.g., Haematopinus eurysternus, Haematopinus suis); Linognathus spp. (e.g., Linognathus vituli); Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes spp., Further examples of pest species that can be controlled by the compounds and / or compositions identified herein include: Phylum Mollusca, Class Bivalvia, e.g., Dreissena spp.; Class Gastropoda, e.g., Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea canaliclata, Succinea spp.; Class helminths, e.g., Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp., Dictyocaulus filaria, Diphyllobothrium latum, Dracunculus medinensis, Echinococcus granulosus, Echinococcus multilocularis, Enterobius vermicularis, Faciola spp., Haemonchus spp. (e.g., Haemonchus contortus); Heterakis spp., Hymenolepis Nanchunga, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca volvulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborni, Strongyloides stercoralis, Strongyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichuria, Wuchereria bancrofti.

[64] 1. A method for determining whether a candidate compound is a modulator of an insect transient receptor potential channel Water witch protein, comprising: (a) providing a first cell expressing the insect transient receptor potential channel Water witch protein; (b) contacting the first cell with the candidate compound; (c) assaying for modulation of the insect transient receptor potential channel Water witch protein, wherein the modulation identifies the candidate compound as a modulator of the insect transient receptor potential channel Water witch protein.

[65] The assaying step comprises: (1) detecting calcium ion mobilization in the first cell in response to the candidate compound; or (2) detecting a membrane potential in the first cell in response to the candidate compound.

[66] The assaying step comprises: (1) comparing calcium ion mobilization in the first cell in the absence of the candidate compound with calcium ion mobilization in the first cell in the presence of the candidate compound; or (2) comparing the membrane potential in the first cell in the absence of the candidate compound with the membrane potential in the first cell in the presence of the candidate compound.

[67] The assaying step comprises: (1) comparing calcium ion mobilization in the first cell in the presence of the candidate compound with a reference level of calcium ion mobilization indicative of no modulation of the insect transient receptor potential channel Water witch protein; or (2) comparing the membrane potential in the first cell in the presence of the candidate compound with a membrane potential reference level indicating no modulation of the insect transient receptor potential channel Water witch protein.

[68] The method according to any one of

[65] to

[67] , wherein the candidate compound regulates the calcium ion mobilization or the membrane potential in the first cell by at least 20% relative to the reference level.

[69] The method according to any one of

[64] to

[67] , wherein the candidate compound is a modulator that inhibits the activity of the insect transient receptor potential channel Water witch protein.

[70] The method according to any one of

[64] to

[67] , wherein the candidate compound is a modulator that activates the insect transient receptor potential channel Water witch protein.

[71] The method according to any one of

[64] to

[67] , wherein modulation of the insect transient receptor potential channel Water witch protein by the candidate compound disrupts insect behavior.

[72] The method according to any one of

[64] to

[67] , wherein modulation of the insect transient receptor potential channel Water witch protein by the candidate compound results in the death of the insect.

[73] The method according to any one of

[64] to

[67] , wherein modulation of the insect transient receptor potential channel Water witch protein by the candidate compound repels insects.

[74] The method according to any one of

[64] to

[67] , wherein the candidate compound is selected from the group consisting of organic small molecules, inorganic small molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

[75] The method according to any one of

[64] to

[67] , wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

[76] Specific species of lepidoptera include Achroia grisella, Acleris spp. (including A. fimbriana, A. gloverana, and A. variana); Acrolepiopsis assectella 、Acronita major 、Adoxophyes spp spp.(from A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp spp. (including A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyresthia conjugella, Argyroploce spp., Argyrotaenia spp. (including A. velutiNanchunga); Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp.、Cacoecia spp.(from C.muriNanchungaC.podana);Cactoblastis cactorum、Cadra cautella、Calingo braziliensis、Caloptilis theivora、Capua reticulana、Carposina spp aridula, Cheimatobia brumata, Chilo spp. (including C. Indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp spp. 、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus);Desmia funeralis, Diaphania spp.hyalinata);Diatraea grandiosella、Diatraea saccharalis、Diphthera festiva、Earias spp.(E.insulana、E.vittella);Ecdytolopha aurantianu、Egira(=Xylomyges)curialis、Elasmopalpus lignosellus、Eldana saccharina、Endopiza viteana、Ennomos subsignaria、Eoreuma loftini、Ephestia spp thrax、Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Faronta albilinea、Feltia spp mellonella、Gracillaria spp.、Grapholite spp.(G.funebrana、G.molesta、G.inopinata); spp. (including H. armigera (=Heliothis armigera) and H. zea (=Heliothis zea)); spp.(excluding H.undalis, H. spp.rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorella, Lerodea eufala, Leucinodes orbonalis, Leucoma salicis, Leucoptera spp. (e.g., L.coffeella, L.scitella); Leuminivora lycinivorell, Lithocolletis blancardella, Lithophane antennata, Llattia octo(=Amyna axis), Lobesia botrana, Lophocampa spp., Loxagrotis albicosta, Loxostege spp. (e.g., L.sticticalis, L.cereralis); Lymantria spp. (e.g., L.dispar, L.monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta);Marasmia spp.Marmara spp.Maruca testulalis、Megalopyge lanata、Melanchra picta、Melanitis leda、Mocis spp separate、Nemapogon cloacella、Neoleucinodes elegantalis、Nepytia spp.、Nymphula spp.、Oiketicus spp.、Omiodes indicata、Omphisa anastomosalis、Operophtera brumata、Orgyia pseudotsugata、Oria spp.Orthaga thyrisalis, Ostrinia spp. (O.nubilalis); Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp licorice, Paysandisia archon, Pectinophora spp., and P. gossypiella; Peridroma saucia, Perileucoptera spp., such as P. coffeella; Phalera bucephala, Phryganidia californica, and Phthorimaea spp. (including P. operculella);Phyllocnistis citrella; Phyllonorycter spp. (including P. blancardella, P. crataegella, P. issikii, P. ringoniella); Pieris spp.(such as P. brassicae, P. rapae, P. napi);Pilocrocis tripunctata, Platypena scabra, Platynota spp.stultana); Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp. (e.g., P.sequax, P.unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana,. Sabulodes aegrotata Schizura concinna Schoenobius spp. Schreckensteinia festaliella Scirpophaga spp spp.(S.inferens、Seudyra subflava、Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.(singえば、T.ni);Night(=Scrobipalpula)absolute、Udea spp.(singhえば、U.rubigalis、U.rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, such as Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A.anxius, A.planipennis, A.sinuatus); Agriotes spp. (e.g. A.fuscicollis, A.lineatus, A.obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A.glabripennis); Anthonomus spp. (e.g. A.eugenii, A.grandis, A.pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A.linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g., B.lentis, B.pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C.assimilis, C.napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E. similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp. (e.g., M. aeneus); Melolontha spp. (e.g., M. hippocastani, M. melolontha); Metamasius hemipterus, Microtheca spp., Migdolus spp. (e.g., M. fryanus), Monochamus spp. (e.g., M. alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P.helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R. billineatus, R. ferrugineus, R. palmarum, R. phoenicis, R. vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S. granaria, S. oryzae, S. zeamais); Sphenophorus spp. (e.g., S. levis); Stegobium paniceum, Sternechus spp. (e.g., S. subsignatus); Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp.、Tenebrio molitor、Tenebrioides mauretanicus、Tribolium spp.(for example、T.castaneum); spp.quadrimaculatus, A.sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. (e.g. C.bezziana, C.hominivorax, C.macellaria); Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp. (e.g. C.hominivorax); Contarinia spp. (e.g. C.sorghicola); Cordylobia anthropophaga, Culex spp. (e.g. 、 C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g. D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g. D. suzukii), Fannia spp. (e.g. F. canicularis); Gastraphilus spp. (e.g. G. intestinalis); Geomyza tipunctata, Glossina spp. (e.g. G. fuscipes, G. morsitans, G. palpalis, G. tachinoides); Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp. (e.g. H. platura); Hypoderma spp.(eg H.lineata);Hyppobosca spp.、Hydrellia philippina、Leptoconops torrens、Liriomyza spp.(eg L.sativae、L.trifolii);Lucilia spp.(eg L.caprina、L.cuprina、L.sericata);Lycoria pectoralis、Mansonia titillanus、Mayetiola spp.(eg M.destructor); spp. (for example, M.autumnalis, M. domestica); Muscina stablens, Oestrus spp. (for example, O. ovis);frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp.; Insects from the order Thysanoptera, e.g., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp. (e.g., F.fusca, F.occidentalis, F.tritici); Heliothrips spp., Hercinothrips femoralis, Kakothrips spp., Microcephalothrips abdominalis, Neohydatothrips samayunkur, Pezothrips kellyanus, Rhipiphorothrips cruentatus, Scirtothrips spp. (e.g., S.citri, S.dorsalis, S.perseae);Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequence、Thrips spp. (including T.imagines, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, and T.tabaci); spp.(such as A.hilare);Acyrthosipon spp.(such as A.onobrychis)A.pisum);Adelges laricis, Adelges tsugae, Adelphocoris spp.(such as A.rapidus,A.superbus);Aeneolamia spp. Agonoscena spp. Aulacorthum solani Aleurocanthus woglumi Aleurodes spp. Aleurodicus disperses Aleurolobus barodensis Aleurothrixus spp. Amrasca spp spp.、Anuraphis cardui、Aonidiella spp.、Aphanostigma pyri、Aphidula nasturtii、Aphis spp.(including A. craccivora, A. fabae, A. forbesi, A. gossypii, A. grossulariae, A. maidiradicis, A. pomi, A. sambuci, A. schneideri, A. spiraecola); Arboridia apicalis, Arilus critatus, Aspidiella spp., Aspidiotus spp., Atanus spp., Aulacaspis yasumatsui, Aulacorthum solani, Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp tabaci));Blissus spp.leucopterus; Brachycaudus spp. (e.g., B. cardui, B. helichrysi, B. persicae, B. prunicola); Brachycolus spp., Brachycorynella asparagi, Brevicoryne brassicae, Cacopsylla spp. (e.g., C. fulguralis, C. pyricola (Psylla piri); Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes ceriferus, Cerosipha gossypii, Chaetosiphon fragaefolii, Chionaspis tegalensis, Chlorita onukii, Chromaphis juglandicola, Chrysomphalus ficus, Cicadulina mbila, Cimex spp. (e.g., C. hemipterus, C. lectularius); Coccomytilus halli, Coccus spp. (e.g., C. hesperidum, C. pseudomagnoliarum); Corythucha arcuata, Creontiades dilutus, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasynus piperis, Dialeurodes spp. (e.g., D. citrifolii); Dalbulus maidis, Diaphorina spp. (e.g., D. citri); Diaspis spp. (e.g., D. bromeliae); Dichelops furcatus, Diconocoris hewetti, Doralis spp.、Dreyfusia nordmannianae、Dreyfusia piceae、Drosicha spp.、Dysaphis spp.(eg D.plantaginea、D.pyri、D.radicola);Dysaulacorthum pseudosolani、Dysdercus spp.(eg D.cingulatus、D.intermedius);Dysmicoccus spp.、Edessa spp、Geocoris spp、Empoasca spp.(eg E.fabae、E.solana);Epidiaspis leperii、Eriosoma spp.(eg E.lanigerum、E.pyricola);Erythroneura spp.、Eurygaster spp.(eg E.integriceps);Euscelis bilobatus、Euschistus spp.(eg E.heros、E.impictiventris、E.servus);Fiorinia theae、Geococcus coffeae、Glycaspis brimblecombei、Halyomorpha spp.(eg H.halys);Heliopeltis spp.、Homalodisca vitripennis (=H.coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp. (e.g. I.purchase); Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp. (e.g. L.ulmi); Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp. (e.g. L.hesperus, L.lineolaris, L.pratensis);. Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp. (e.g. M.rosae, M.avenae, M.euphorbiae); Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (=Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis coryli, Murgantia spp., Myzus spp. (e.g. M.ascalonicus, M.cerasi, M.nicotianae, M.persicae, M.varians); Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp. (e.g. N.malayanus, N.nigropictus, N.parvus, N.virescens); Nezara spp.(eg N.viridula);Nilaparvata lugens、Nysius huttoni、Oebalus spp.(eg O.pugnax);Oncometopia spp.、Orthezia praelonga、Oxycaraenus hyalinipennis、Parabemisia myricae、Parlatoria spp.、Parthenolecanium spp.(eg、P.corni、P.persicae);Pemphigus spp.(eg、P.bursarius、P.populivenae);Peregrinus maidis、Perkinsiella saccharicida、Phenacoccus spp. (for example、P. aceris、P.gossypii); Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp. (e.g. P.devastatrix), Piesma quadrata, Piezodorus spp. (e.g. P.guildinii); Pinnaspis aspidistrae, Planococcus spp. (e.g. P.citri, P.ficus); Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudacysta persea, Pseudaulacaspis pentagona, Pseudoococcus spp. (e.g. P.comstocki); Psylla spp. (e.g. P.mali); Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp. (e.g. Q.perniciosus); Quesada gigas, Rastroccus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp. (e.g., R. pseudobrassicas, R. insertum, R. maidis, R. padi); Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulatus, Sitobion avenae, Sogata spp.、Sogatella furcifera、Solubea insularis、Spissistilus festinus(=Stictocephala festina); spp.(including T.accerra;T.perditor);Tibraca spp., Tomaspis spp., Toxoptera spp.(including T.aurantii);Trialeurodes spp.(including T.abutilonea, T.ricini, and T.vaporariorum);Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp.(including U.citri,U.yanonensis);Viteus vitifolii;The list of Hymenoptera includes Acanthomyops interjectus, Athalia rosae, Atta spp (including A. capiguara, A. cephalotes, A. cephalotes, A. laevigata, A. robusta, A. sexdens, A. texana), Bombus spp., Brachymyrmex spp., and Camponotus spp (from C. floridanus, C. pennsylvanicus, C. modoc); Cardiocondyla nuda, Chalibion ​​sp, Crematogaster spp, Dasymutilla occidentalis, Diprion spp, Dolichovespula maculata, Dorymyrmex spp, Dryocosmus kuriphilus, Formica spp, Hoplocampa spp., (excluding H.minuta, H. testudinea); Iridomyrmex humilis, Lasius spp.niger) Linepithema humile Liometopum spp Leptocybe invasa Monomorium spp(M.pharaonis) Monomorium Nylandria fulva Pachycondyla chinensis Paratrechina longicornis Paravespula spp(such as P. germanica, P. pennsylvanica, P. vulgaris);Pheidole spp(such as P. megacephala);Pogonomyrmex spp(such as P. barbatus, P. californicus), Polistes rubiginosa、Prenolepis impairs、Pseudomyrmex gracilis、Schelipron spp、Sirex cyaneus、Solenopsis spp(S.geminata、S.invicta、S.molesta、S.richteri、S.xyloni) Sphecius speciosus spp (including V.squamosal);Wasmannia auropunctata, Xylocopa sp.;Orthoptera species include Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, and Ceuthophilus spp. Diastrammena asynamora. Dociostaurus maroccanus spp.(such as L. migratoria, L. pardalina);Melanoplus spp.(such as M. spp.).bivittatus, M. femurrubrum, M. mexicanus, M. sanguinipes, M. spretus); spp(S.americana, S.gregaria), Stemopelmatus spp, Tachycines asynamorus, and Zonozerus variegatus;Arachnidae, Argasidae, Ixodidae, Sarcoptidae (Acari), Amblyomma spp.(A.americanum, A.variegatum, A.maculatum), Argas spp., (A. persicu), Boophilus spp., B. annulatus, B. decoloratus, B. microplus) Dermacentor spp (including D. silvarum, D. andersoni, D. variabilis), Hyalomma spp. (including H. truncatum), Ixodes spp.(I.ricinus, I.rubicundus, I.scapularis, I.holocyclus, I.pacificus) Rhipicephalus sanguineus Ornithodorus spp.(excluding O. moubata, O. hermsi, O. turicata), Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp., (excluding P.ovis), Rhipicephalus spp(including R.sanguineus, R.appendiculatus), Rhipicephalus evertsi), Rhizoglyphus spp; Sarcoptes spp. (including S. Scabiei); and Aceria spp (including A. sheldoni, A. anthocoptes), Acallitus spp; Aculops spp.pelekassi); Aculus spp. (e.g., A. schlechtendali); Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; the Eriophyidae family, including Eriophytes ribis and Eriophyes spp. (e.g., Eriophyes sheldoni); the Tarsonemiidae family, including Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp., and Stenotarsonemus spinki; the Tenuipalpidae family, including Brevipalpus spp. (e.g., B. phoenicis); Eotetranychus spp., Eut. The Tetranychidae family includes species such as etranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp. (e.g., T. cinnabarinus, T. evansi, T. kanzawai, T. pacificus, T. phaseulus, T. telarius, and T. urticae); Bryobia praetiosa; Panonychus spp. (e.g., P. ulmi, P. citri); Metatetranychus spp., and Oligonychus spp. (e.g., O. pratensis, O. perseae); Vasates lycopersici; Raoiella indica; Carpoglyphidae, including Carpoglyphus spp.; Penthaleidae spp. (e.g., Halotydeus destructor); Demodicidae, including species such as Demodex spp.; Trombicula family Trombicidea, including spp.; family Macronyssidae, including Ornothonyssus spp.; family Pyemotidae, including Pyemotes tritici; Tyrophagus putrescentiae; family Acaridae, including Acarus siro; family Araneida, including Latrodectus mactans, Tegenaria agrestis, Chiracanthium spp., Lycosa spp., Achaearanea tepidariorum, and Loxosceles reclusa; pests from the phylum Nematoda, for example, plant-parasitic nematodes (e.g., root-knot nematodes) Meloidogyne spp. (e.g., M. hapla, M. incognita, M. javanica); cyst nematodes Globodera spp. (e.g., G. rostochiensis); Heterodera (e.g. H. avenae, H. glycines, H. schachtii, H. trifolii); Seed gall nematodes Anguina spp.Stem and foliar nematodes Aphelenchoides spp. (e.g., A. besseyi); Sting nematodes Belonolaimus spp. (e.g., B. longicaudatus); Pine nematodes Bursaphelenchus spp. (e.g., B. lignicolus, B. xylophilus); Ring nematodes Criconema spp.; Criconemella spp. (e.g., C. xenoplax and C. ornata); and Criconemoides spp. (e.g., Criconemoides informis; Mesocriconema spp.); Stem and bulb nematodes Ditylenchus spp. (e.g., D. destructor, D. dipsaci); Awl nematodes (Dolichodorus spp.); spiral nematodes (Heliocotylenchus multicinctus); sheath and sheathoid nematodes (Hemicycliophora spp. and Hemicriconemoides spp.); Hirshmanniella spp.; lance nematodes (Hoploaimus spp.); false rootknot nematodes (Nacobbus spp.); needle nematodes (Longidorus spp. (e.g., L. elongatus); root-lesion nematodes (Pratylenchus spp. (e.g., P. brachyurus, P. neglectus, P. penetrans, P. curvitatus, P. goodeyi); Burrowing nematodes Radopholus spp. (e.g., R. similis; Rhadopholus spp.; Rhodopholus spp.;Reniform nematodes Rotylenchus spp. (e.g., R. robustus, R. reniformis); Scutellonema spp.;Stubby-root nematodes Trichodorus spp. (e.g., T. obtusus, T. primitivus); Paratrichodorus spp. (e.g., P. minor);Stunt nematodes Tylenchorhynchus spp. (e.g., T. claytoni, T. dubius);Citrus nematodes Tylenchulus spp. (e.g., T. semipenetrans);Dagger nematodes Xiphinema spp.;and other plant-parasitic nematode species;Insecta from the order Isoptera, for example, Calotermes flavicollis, Coptotermes spp (e.g. C. formosanus, C. gestroi, C. acinaciformis); Cornitermes cumulans, Cryptotermes spp (e.g. C. brevis, C. cavifrons); Globitermes sulfureus, Heterotermes spp (e.g. H. aureus, H. longiceps, H. tenuis); Leucotermes flavipes, Odontotermes spp., Incisitermes spp (e.g. I. minor, I. Snyder); Marginitermes hubbardi, Mastotermes spp (e.g. M. darwiniensis), Neocapritermes spp (e.g. N.opacus, N.parvus); Neotermes spp, Procornitermes spp, Zootermopsis spp. (e.g. Z. angusticollis, Z. nevadensis), Reticulitermes spp. (e.g. R. hesperus, R. tibialis, R. speratus, R. flavipes, R. grassei, R. lucifugus, R.santonensis, R.virginicus);Termes natalensis;Insects from the order Blattaria, such as Blatta spp (e.g. B.orientalis, B.lateralis);Blattella spp (e.g. B.asahinae, B.germanica);Leucophaea maderae, Panchlora nivea, Periplaneta spp (e.g. P.americana, P.australasiae, P.brunnea, P.fuligginosa, P.japonica); Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnoscelus surinamensis; insects from the order Siphonoptera, e.g. Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp (e.g. C. felis, C. canis), Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus; insects from the order Thysanura, for example, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica; pests from the class Chilopoda, for example, Geophilus spp., Scutigera spp. (e.g., Scutigera coleoptrata); pests from the class Diplopoda, for example, Blaniulus guttulatus, Julus spp., Narceus spp.; pests from the class Symphyla, for example, Scutigerella immaculata; insects from the order Dermaptera, for example, Forficula auricularia; insects from the order Collembola, for example, Onychiurus spp.(e.g., Onychiurus armatus); pests from the order Isopoda, for example, Armadillidium vulgare, Oniscus asellus, Porcellio scaber; insects from the order Phthiraptera, for example, Damalinia spp., Pediculus spp. (e.g., Pediculus humanus capitis, Pediculus humanus corporis, Pediculus humanus humanus); Pthirus pubis, Haematopinus spp. (e.g., Haematopinus eurysternus, Haematopinus suis); Linognathus spp. (e.g., Linognathus vituli); Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes Examples of further pest species that may be controlled by the compounds and / or compositions identified herein include the phylum Mollusca, the class Bivalvia, e.g., Dreissena spp.; the class Gastropoda, e.g., Arion spp., Biomphalaria spp., Bulinus spp., Deroceras spp., Galba spp., Lymnaea spp., Oncomelania spp., Pomacea canaliclata, Succinea spp.; the class Helminthi, e.g., Ancylostoma duodenale, Ancylostoma ceylanicum, Acylostoma braziliensis, Ancylostoma spp., Ascaris lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp., Dictyocaulus filaria, Diphyllobothrium latum, Dracunculus medinensis, Echinococcus granulosus, Echinococcus multilocularis, Enterobius vermicularis, Faciola spp., Haemonchus spp. (e.g., Haemonchus contortus); Heterakis spp., Hymenolepis Nanchunga, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca vo. lv ulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborni, Strongyloides stercora lis, Stronyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichuria, and Wuchereria bancrofti.

[77] A candidate compound selected by the method according to any one of

[64] to

[67] .

[78] The method according to

[77] , wherein the candidate compound is a modulator that inhibits the activity of the insect transient receptor potential channel Water witch protein.

[79] The method according to

[77] , wherein the candidate compound is a modulator that activates the insect transient receptor potential channel Water witch protein.

[80] The candidate compound according to

[78] or

[79] , wherein the candidate compound interferes with insect behavior.

[81] The candidate compound according to

[78] or

[79] , wherein the candidate compound kills insects.

[82] The candidate compound according to

[78] or

[79] , wherein the candidate compound repels insects.

[83] A method for controlling insects, comprising applying to insects a candidate compound selected by the method according to

[78] or

[79] .

[84] The method according to

[83] , wherein the candidate compound is an inhibitor of the insect transient receptor potential channel Water witch protein.

[85] The method according to

[83] , wherein the candidate compound is an activator of the insect transient receptor potential channel Water witch protein.

[86] The method according to any one of

[83] to

[85] , wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

[87] Specific species of lepidoptera include Achroia grisella, Acleris spp. (including A. fimbriana, A. gloverana, and A. variana); Acrolepiopsis assectella 、Acronita major 、Adoxophyes spp spp.(from A. exclamationis, A. fucosa, A. ipsilon, A. orthogoma, A. segetum, A. subterranea); transitella, Anacampsis sarcitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp spp. (including A. argyrospila, A. fuscocupreanus, A. rosana, A. xyloseanus); Argyresthia conjugella, Argyroploce spp., Argyrotaenia spp. (including A. velutiNanchunga); Athetis mindara、Austroasca viridigrisea、Autographa gamma、Autographa nigrisigna、Barathra brassicae、Bedellia spp.、Bonagota salubricola、Borbo cinnara、Bucculatrix thurberiella、Bupalus piniarius、Busseola spp.、Cacoecia spp.(from C.muriNanchungaC.podana);Cactoblastis cactorum、Cadra cautella、Calingo braziliensis、Caloptilis theivora、Capua reticulana、Carposina spp aridula, Cheimatobia brumata, Chilo spp. (including C. Indicus, C. suppressalis, C. partellus); Choreutis pariana, Choristoneura spp.(C.conflictana, C. fumiferana, C. longicellana, C. muriNanchunga, C. occidentalis 、C.rosaceana);Chrysodeixis(=Pseudoplusia)spp.(including C.eriosoma、C.includens);Cirphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp spp. 、Corcyra cephalonica、Crambus caliginosellus、Crambus teterrellus, Crocidosema (= Epinotia) aporema, Cydalima (= Diaphania) perspectalis, Cydia (= Carpocapsa) spp pinicola, Dendrolimus spp. (including D. pini, D. spectabilis, D. sibiricus);Desmia funeralis, Diaphania spp.hyalinata);Diatraea grandiosella、Diatraea saccharalis、Diphthera festiva、Earias spp.(E.insulana、E.vittella);Ecdytolopha aurantianu、Egira(=Xylomyges)curialis、Elasmopalpus lignosellus、Eldana saccharina、Endopiza viteana、Ennomos subsignaria、Eoreuma loftini、Ephestia spp thrax、Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Faronta albilinea、Feltia spp mellonella、Gracillaria spp.、Grapholite spp.(G.funebrana、G.molesta、G.inopinata); spp. (including H. armigera (=Heliothis armigera) and H. zea (=Heliothis zea)); spp.(excluding H.undalis, H. spp.rogatalis); Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magNanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta malinellus, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorella, Lerodea eufala, Leucinodes orbonalis, Leucoma salicis, Leucoptera spp. (e.g., L.coffeella, L.scitella); Leuminivora lycinivorell, Lithocolletis blancardella, Lithophane antennata, Llattia octo(=Amyna axis), Lobesia botrana, Lophocampa spp., Loxagrotis albicosta, Loxostege spp. (e.g., L.sticticalis, L.cereralis); Lymantria spp. (e.g., L.dispar, L.monacha); Lyonetia clerkella, Lyonetia prunifoliella, Malacosoma spp. (e.g., M.americanum, M.californicum, M.constrictum, M.neustria); Mamestra spp. (e.g., M.brassicae, M.configurata); Mamestra brassicae, Manduca spp. (e.g., M.quinquemaculata, M.sexta);Marasmia spp.Marmara spp.Maruca testulalis、Megalopyge lanata、Melanchra picta、Melanitis leda、Mocis spp separate、Nemapogon cloacella、Neoleucinodes elegantalis、Nepytia spp.、Nymphula spp.、Oiketicus spp.、Omiodes indicata、Omphisa anastomosalis、Operophtera brumata、Orgyia pseudotsugata、Oria spp.Orthaga thyrisalis, Ostrinia spp. (O.nubilalis); Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp licorice, Paysandisia archon, Pectinophora spp., and P. gossypiella; Peridroma saucia, Perileucoptera spp., such as P. coffeella; Phalera bucephala, Phryganidia californica, and Phthorimaea spp. (including P. operculella);Phyllocnistis citrella; Phyllonorycter spp. (including P. blancardella, P. crataegella, P. issikii, P. ringoniella); Pieris spp.(such as P. brassicae, P. rapae, P. napi);Pilocrocis tripunctata, Platypena scabra, Platynota spp.stultana); Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp. (e.g., P.sequax, P.unipuncta); Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana,. Sabulodes aegrotata Schizura concinna Schoenobius spp. Schreckensteinia festaliella Scirpophaga spp spp.(S.inferens、Seudyra subflava、Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis spp. Stomopteryx subsecivella Strymon bazochii Sylepta derogata SyNanchungthedon spp. (S. exitiosa) Tecia solanivora Telehin licus Thaumatopoea pityocampa、Thaumatotibia(=Cryptophlebia)leucotreta、Thaumetopoea pityocampa、Thecla spp.、Theresimima ampelophaga、Thyrinteina spp、Tildenia inconspicuella、Tinea spp. (including T. cloacella and T. pellionella); Tineola bisselliella and Tortrix spp. (including T. viridana); Trichophaga tapetzella and Trichoplusia spp.(singえば、T.ni);Night(=Scrobipalpula)absolute、Udea spp.(singhえば、U.rubigalis、U.rubigalis); Virachola spp., Yponomeuta padella, and Zeiraphera canadensis; insects from the order Coleoptera, such as Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp. (e.g. A.anxius, A.planipennis, A.sinuatus); Agriotes spp. (e.g. A.fuscicollis, A.lineatus, A.obscurus); Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. (e.g. A.glabripennis); Anthonomus spp. (e.g. A.eugenii, A.grandis, A.pomorum); Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp. (e.g. A.linearis); Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. (e.g., B.lentis, B.pisorum, B.rufimanus); Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp. (e.g., C.assimilis, C.napi); Chaetocnema tibialis, Cleonus mendicus, Conoderus spp. (e.g., C. vespertinus); Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagi, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp. (e.g., C. destructor); Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp. (e.g., D. undecimpunctata, D. speciosa, D. longicornis, D. semipunctata, D. virgifera); Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp. (e.g., E. varivestis, E. vigintioctomaculata); Epitrix spp. (e.g., E. hirtipennis, E. similaris); Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp. (e.g., H. brunneipennis, H. postica); Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp. (e.g., L. bilineata, L. melanopus); Leptinotarsa ​​spp. (e.g., L. decemlineata); Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp. (e.g., L. bruneus); Liogenys fuscus, Macrodactylus spp. (e.g., M. subspinosus); Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp. (e.g., M. aeneus); Melolontha spp. (e.g., M. hippocastani, M. melolontha); Metamasius hemipterus, Microtheca spp., Migdolus spp. (e.g., M. fryanus), Monochamus spp. (e.g., M. alternatus); Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorrhynchus sulcatus, Otiorrhynchus ovatus, Otiorrhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp. (e.g., P. brassicae, P. cochleariae); Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp. (e.g., P.helleri); Phyllotreta spp. (e.g., P. chrysocephala, P. nemorum, P. striolata, P. vittula); Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp. (e.g., R. billineatus, R. ferrugineus, R. palmarum, R. phoenicis, R. vulneratus); Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp. (e.g., S. granaria, S. oryzae, S. zeamais); Sphenophorus spp. (e.g., S. levis); Stegobium paniceum, Sternechus spp. (e.g., S. subsignatus); Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp.、Tenebrio molitor、Tenebrioides mauretanicus、Tribolium spp.(for example、T.castaneum); spp.quadrimaculatus, A.sinensis); Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp. (e.g. C.bezziana, C.hominivorax, C.macellaria); Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp. (e.g. C.hominivorax); Contarinia spp. (e.g. C.sorghicola); Cordylobia anthropophaga, Culex spp. (e.g. 、 C. nigripalpus, C. pipiens, C. quinquefasciatus, C. tarsalis, C. tritaeniorhynchus); Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp. (e.g. D. antique, D. coarctata, D. platura, D. radicum); Dermatobia hominis, Drosophila spp. (e.g. D. suzukii), Fannia spp. (e.g. F. canicularis); Gastraphilus spp. (e.g. G. intestinalis); Geomyza tipunctata, Glossina spp. (e.g. G. fuscipes, G. morsitans, G. palpalis, G. tachinoides); Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp. (e.g. H. platura); Hypoderma spp.(eg H.lineata);Hyppobosca spp.、Hydrellia philippina、Leptoconops torrens、Liriomyza spp.(eg L.sativae、L.trifolii);Lucilia spp.(eg L.caprina、L.cuprina、L.sericata);Lycoria pectoralis、Mansonia titillanus、Mayetiola spp.(eg M.destructor); spp. (for example, M.autumnalis, M. domestica); Muscina stablens, Oestrus spp. (for example, O. ovis);frit); Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp. (e.g., P.antiqua, P.brassicae, P.coarctata); Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp. (e.g., R.cerasi, R.cingulate, R.indifferens, R.mendax, R.pomonella); Rivellia quadrifasciata, Sarcophaga spp. (e.g., S.haemorrhoidalis); Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp. (e.g., S.calcitrans); Tabanus spp. (e.g., T.atratus, T.bovinus, T.lineola, T.similis); Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp.; Insects from the order Thysanoptera, e.g., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp. (e.g., F.fusca, F.occidentalis, F.tritici); Heliothrips spp., Hercinothrips femoralis, Kakothrips spp., Microcephalothrips abdominalis, Neohydatothrips samayunkur, Pezothrips kellyanus, Rhipiphorothrips cruentatus, Scirtothrips spp. (e.g., S.citri, S.dorsalis, S.perseae);Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequence、Thrips spp. (including T.imagines, T.hawaiiensis, T.oryzae, T.palmi, T.parvispinus, and T.tabaci); spp.(such as A.hilare);Acyrthosipon spp.(such as A.onobrychis)A.pisum);Adelges laricis, ...

Claims

1. 1. A method for determining whether a candidate compound is a modulator of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein, comprising: (a) providing a first cell that co-expresses the insect transient receptor potential channel Nanchung protein and the insect transient receptor potential channel Water witch protein; (b) contacting the first cell with the candidate compound; (c) assaying for modulation of said complex, wherein based on said modulation, said candidate compound is identified as a modulator of said complex; The assaying step of (c) comprises: (1) detecting calcium ion mobilization in the first cell in response to the candidate compound; or (2) detecting a membrane potential in the first cell in response to the candidate compound; The method includes at least one of the following:

2. The assaying step comprises the steps of: (1) comparing calcium ion mobilization in the first cell in the absence of the candidate compound with calcium ion mobilization in the first cell in the presence of the candidate compound; or (2) comparing the membrane potential in the first cell in the absence of the candidate compound with the membrane potential in the first cell in the presence of the candidate compound.

3. The assaying step comprises the steps of: (1) comparing calcium ion mobilization in the first cell in the presence of the candidate compound with a reference level of calcium ion mobilization indicative of no modulation of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein; or (2) comparing the membrane potential in the first cell in the presence of the candidate compound with a membrane potential reference level indicative of no modulation of a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein.

4. The method of claim 3, wherein the candidate compound modulates the calcium ion mobilization or the membrane potential in the first cell by at least 20% relative to the reference level.

5. The method of any one of claims 1 to 3, wherein the candidate compound is a modulator that inhibits the activity of the complex.

6. The method of any one of claims 1 to 3, wherein the candidate compound is a modulator that activates the complex.

7. The method of any one of claims 1 to 3, wherein modulation of the complex by the candidate compound disrupts insect behaviour.

8. The method of any one of claims 1 to 3, wherein modulation of the complex by the candidate compound results in killing of the insect.

9. The method of any one of claims 1 to 3, wherein modulation of the complex by the candidate compound repels insects.

10. The method of any one of claims 1 to 3, wherein the candidate compound is selected from the group consisting of small organic molecules, small inorganic molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

11. The method according to any one of claims 1 to 3, wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

12. The insects are selected from the group consisting of insects from the order Lepidoptera, insects from the order Coleoptera, insects from the order Diptera, insects from the order Thysanoptera, insects from the order Hemiptera, insects from the order Hymenoptera, insects from the order Orthoptera, pests from the class Arachnida, pests from the phylum Nematoda, insects from the order Isoptera, insects from the order Blattaria, insects from the order Siphonoptera, insects from the order T 4. The method according to any one of claims 1 to 3, wherein the insects are selected from the group consisting of insects from the order Hysanura, pests from the class Chilopoda, pests from the class Diplopoda, pests from the class Symphyla, insects from the order Dermaptera, insects from the order Collembola, pests from the order Isopoda, pests from the phylum Mollusca, pests from the class Bivalvia, pests from the class Gastropoda and pests from the class Helminthi. [13] The aforementioned insects, Achroia grisella, Acleris spp. argiellacea、Aerrodicus dispersus、Alsophila pômetaria、Ambelophaga rubiginosa、Ammelois transitella、Anacampsis sarcitella、Anagasta Kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp.; Apamea spp., Aproaerema modicella, Archips spp.; Argyrestia conjugella, Argyroploce spp., Argyrotaenia spp.; Athetis mindara, Austriasca viridigrisea, Autographa gamma, Autographa nigrisigna, Barathra brassicae, Bedelia spp., Bonagota salubricola, Borbo cinnara, Bucculatrix thurberiella, Bupalus piniarius, Busseola spp., Cacoecia spp.; Cactoblastis cactorum, Cadra cautiona, Calingo braziliensis, Caloptilis theivora, Capua reticulana、Carpōsina spp.;Cephus spp、Chaetocnema aridula、Cheimatobia brumata、Chilo spp.;Choreutis pariana、Choristoneura spp;Cyrphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp., Colias eurytheme, Conopomorpha spp., Conotrachelus spp., Copitarsia spp., Corcyra cephalonica, Crambus caliginosellus, Crambus Teterrellus, Crocidosema (=Epinotia) aporema, Cydalima (=Diaphania) perspectalis, Cydia (=Carpocapsa) spp.; Dalaca noctuides, Datana integrrima, Dasychira pinicola, Dendrolimus spp.; Desmia funeralis, Diaphania spp.; Diatraea grandiosella, Diatraea saccharalis, Diphthera Festiva, Earias spp.; Ecdytolopha aurantianu, Egira (= Xylomyges) curialis, Elasmopalpus lignosellus, Eldana saccharina, Endopiza vitiana, Ennomos subsignaria, Eoreuma loftini, Ephestia spp.; Epinotia aporema, Epiphyas postvittana, Eranis tiliaria, Erionota thrax, Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Farona albilinea、Feltia spp.;Galleria m---;;;racillaria spp.;Grapholita spp.;Halysidota spp.;Harrisina americana;Hedylepta spp.;Helicoverpa spp;Heliothis spp.; Hellula spp.; Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magnanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta Malinella, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina ficellaria ficellaria, Lambdina ficellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorell, Lerodea euphala, Leucinodes orbonalis, Leucoma salicis, Leucoptera Ap;Leemmmmvora ]ymmgvorellstitinetineneeeeeeeereierereieieeieereereeieiiiteeeneeeererererer botrana、Lootcampa spp;;tagrotis albicosta;tostege spp;;ymmantria spp;;yonetia clerkella;yonetia #renfoliell;Mesoooma ;Mmestra spp.;Mastra brassicae, Manduca spp.;Marasmia spp., Marmara spp., Maruca testulalis, Megalopyge lanata, Melanchra picta, Melanitis leda, Mocis spp.;Mocis latipes, Monochroa fragariae, Mythimna separatea, Nemapogon cloacella, Neoleucinodes elegantis, Nepytia spp., Nymphula spp., Oiketicus spp., Omiodes indicata, Omphisa anastomosalis, Operophtera brumata, Orgyia pseudotsugata, Oria spp., Orthaga Thyrisalis, Ostrinia spp.; Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp., Papaipema nebris, Papilio cresphontes, Paramyelois transitella, Paranthrene regalis, Paysandisia archon, Pectinophora spp.; Peridroma saucia, Perileucoptera spp.; Phalera Bucephala, Phryganidia california, Phthorimaea spp.; Phyllocnistis citrella, Phyllonorycter spp.; Pieris spp.; Pilocrocis tripunctata, Platypena scabra, Platynota spp.; Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp., Plutella Maculopennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigon, Pseudaletia spp.;Pyrausta nubilalis, Rachyplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana, Sabulodes aegrotata, Schizura concinna, Schoenobius spp., Schreckensteinia festaliella, Scirpophaga spp.; Scotia segetum, Sesamia spp., Seudyra subflava, Sitotroga cerealella、Sparganothis pilleriana、Spilonota lechriaspis、S. ocellana、Spodoptera(==LLTygmma)spp.;Stigmella spp.、Stomopteryx subsecivella、Strymmon bazochii、Sylepta derogata、SyNachungthedon spp.、Tecia solanivora、Telehin licus、Taumatopoea ღანამებისება, ღამომებისამები (==Cryაღაღღღამება) leucotreta amrlopphaga、Tyrinteina spp;yldenia inconspicuella、yinea spp.;yoneola bissellimtyyyyyy spp.; Trichophagaパテウ゚ella、yrichôoslsia spp.; Tuta (= Scrobipalpula) absolute, Udea spp.; Virachola spp., Yponomeuta padella, and Zeiraphera canadensis, Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp.; Agriotes spp.; Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp. ;Anthonomus spp. ; Anthrenus spp. , Aphthona euphoridae, Apion spp. , Apogonia spp. , Atous haemorrhoidalis, Atomaria spp. ; Attagenus spp. , Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp. ; Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchu s spp.; Chaetocnema tibialis, Cleonus mendicus, Conoderus spp.; Conotrachelus nenuphar, Cosmopolites spp., Costelytra zealandica, Crioceris asparagus, Cryptolestes ferrugineus, Cryptorhynchus lapathi, Ctenicera spp.; Curculio spp., Cylindrocopterus spp., Cyclocephala spp, Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp.; Diaprepes abbreviates, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp.; Epitrix spp.; Eutheola humilis, Eutinobothrus braziliensis, Faustinus cubae, Gibbium psylloids, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp.; Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp.; Leptinotarsa ​​spp.; Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp.;Liogenys fuscus, Macrodactylus spp.; Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp.; Melolontha spp.; Metamasius hemipterus, Microtheca spp., Migdolus spp., Monochamus spp.; Naupactus xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorhynchus sulcatus, Otiorhynchus ovatus, Otiorhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp.; Phoracantha recurva, Phyllobius pyri, Phyllopertha horticola, Phyllophaga spp.; Phyllotreta spp.; Phyllopertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp.; Saperda candida, Scolytus schevyrewi, Scyphophorus acupunctatus, Sitona lineatus, Sitophilus spp.; Sphenophorus spp.; Stegobium paniceum, Sternechus spp.;Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp., Tenebrio molitor, Tenebrioides mauretanicus, Tribolium spp.; Trogoderma spp., Tychius spp., Xylotrechus spp.; and Zabrus spp., Aedes spp.; Anastrepha ludens, Anopheles spp.; Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomya spp.; Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp.; Contarinia spp.; Cordylobia anthropophaga, Culex spp.; Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp.; Dermatobia hominis, Drosophila spp., Fannia spp.; Gastraphilus spp.; Geomyza tipunctata, Glossina spp.; Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp.; Hypoderma spp.; Hippobosca spp., Hydrellia philippina, Leptoconops torrens, Liriomyza spp.; Lucilia spp.; Lycoria pectoralis, Mansonia titillanus, Mayetiola spp.; Musca spp.; Muscina stablens, Oestrus spp.;Opomyza florum, Oscinella spp.; Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp.; Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp.; Rivellia quadrifasciata, Sarcophaga spp.; Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp.; Tabanus spp.; Tannia spp., Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus、･eeeothripsfllavens、Frankliniella spp.;Heliothrips spp.、Hercinothrips fmmoralis、Kakothrips spp.、MAoce.stttetteteteteeteteterer cruentatus、Scirtothrips spp.;Stenchaetothrips spp, Taeniothrips cardamoni, Taeniothrips inconsequens, Thrips spp., Acizzia jamatonica, Acrosternum spp.; Acyrthosipon spp.; Adelges laricis, Adelges tsugae, Adelphocoris spp.;Aeneolamia spp., Agonoscena spp., Aulacorthum solani, Aleuracanthus woglumi, Aleurodes spp., Aleurodicus disperses, Aleurolobus barodensis, Aleurothrixus spp., Amrasca spp., Anasa tristis, Antestiopsis spp., Anuraphis cardui, Aonidella spp., Aphanostigma ;;;;;;;;Arboridia apicalis;Arilus critatus;Aspidiella spp.;Aspidiotus spp.;Atanus spp.、Aulacaspis yasumatsui、Aulacorthum solani、Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp.;Blissus tail spp.; Brachycolus spp., Brachycorynella asparagus, Brevicoryne brassicae, Cacopsylla spp.; Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes cerifferus、Cerosipha gossypii、Chaetosiphon fragaefolii、Chionaspis tegalensis、Chlorita onukii、Chromaphis juglandicola、;yoomｾaaffaoorionionionionmmimmmmmimmmmmem spp.; Coccomytilus halli; Coccos spp.Corythucha arcuata, Creontiades dilutes, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasinus piperis, Dialeurodes spp.; Dalbulus maidis, Diaphorina spp.; Diaspis spp.; Dichelops Furcatus, Diconocoris hewettii, Doralis spp., Dreyfusia nordmannianae, Dreyfusia piceae, Drosicha spp., Dysaphis spp.; Dysaulacorthum pseudosolani, Dysdercus spp.; Dysmicoccus spp., Edessa spp., Geocoris spp., Empoasca spp.; Epidiaspis leperii, Eriosoma spp.; Erythroneura spp., Eurygaster spp.; Euscelis bilobatus, Euschistus spp.; Fiorinia theae, Geococcus coffeae, Glycaspis brimblecombei, Halyomorpha spp.; Heliopeltis spp., Homalodisca vitripennis (= H. coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus lactucae, Icerya spp.; Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp.; Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp.; Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp.;Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megoura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (= Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis Corylia, Murgantia spp., Myzus spp.; Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp.; Nezara spp.; Nilaparvata lugens, Nysius huttoni, Oebalus spp.; Oncometopia spp., Orthozia praelonga, Oxycaraenus hyalinipennis, Parabemisia Myricae, Parlatoria spp., Parthenolecanium spp.; Pemphigus spp.; Peregrinus maidis, Perkinsiella saccharicida, Phenacococcus spp.; Phloeomyzus passerinii, Phorodon humuli, Phylloxera spp., Piesma quadrata, Piezodorus spp.; Pinnaspis aspidistrae, Planococcus spp.; Prosapias bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudocysta persea, Pseudoulacaspis pentagona, Pseudococcos spp.; Psylla spp.; Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp.;Quesada gigas、Astrócoccus spp.、Reduvius senilis、Rhizoecus americanus、Rhodnius spp.、Rhopalomyzus ascalonicus、Rhopalosiphum spp.;Sagatodes spp.、Sahlbergella singularis、Saissetia spp.、Sappaphis mala、Sappaphis m--、Scaptocoris spp、Scaphoides titanus, Schizophis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulates, Sitobion avenae, Sogata spp., Sogatella furcifera, Solubea insularis, Spissistylus festinus (= Stictocephala festina); Stephanitis nashi, Stephanitis pyrioides, Stephanitis takeyai、fenalaphara fmalayensis、ftetraleurodes perseae、ftherioaphis fmaculate、fyanta spp.;fbraca spp.、Tomaspis spp.、Toxoptera spp;;TAiSeuroodes s@;;Triatoma s@;;TO:a s@.;TEElocyba s@;;U.aspis s@; and Viteus vitiffolii、Acanthomyops interjectus、Athalia rosae, Atta spp, Bombus spp., Brachymyrmex spp., Camponotus spp.; Cardiocondyla nuda, Chalibion ​​sp., Crematogaster spp., Dasymutilla occidentalis, Diprion spp., Dolichovespula maculata, Dorymyrex spp., Dryocosmus kuriphilus, Formica spp., Hoplocampa spp.;Iridomyrme humilis, Lasius spp., Linepithema humile, Liometopum spp., Leptocybe invasa, Monomorium spp., Monomorium, Nylandria fulva, Pachycondyla chinensis, Paratrechina longicornis, Paravespula spp.; Pheidole spp.; Pogonomyrme spp., Polystes rubiginosa, Prenolepis Impairs, Pseudomyrmex gracilis, Schelipron spp, Sirex cyaneus, Solenopsis spp, Sphecius speciosus, Sphex spp, Tapinoma spp; Tetramorium spp, Vespa spp; Vespula spp; Wasmannia auropunctata, Xylocopa spp; Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, Ceutophilus spp, Diastrammena asynamora, Dociostaurus maroccanus, Gryllotalpa spp; Gryllus spp, Hieroglyphus daganensis, Kraussaria angulifera, Locusta spp; Melanoplus spp; Nomadacris septemfasciata, Oedaleus senegalensis, Scapteriscus spp, Schistocerca spp., Stemopelmmatus spp.Acari of the families Argasidae, Ixodidae and Sarcoptidae, Amblyomma spp. , Argas spp. , Boophilus spp. , Dermacentor spp, Hyalomma spp. , Ixodes spp. , Rhipicephalus sanguineus, Ornithodorus spp. , Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp, Rhipicephalus spp, Rhizoglyphus spp; Sarcoptes spp. ; and Aceria spp, Acallitus spp; Aculops spp. ; Aculus spp.; Colomerus vitis, Epitrimerus pyri, Phyllocoptruta oleivora; Eriophyidae family including Eriophytes ribis and Eriophyes spp.; Hemitarsonemus spp., Phytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp.; Tarsonemiidae family including Stenotarsonemus spinki; Brevipalpus spp. Family Tenuipalpidae, including Eotetranychus spp. , Eutetranychus spp. , Oligonychus spp. , Petrobia latens, Tetranychus spp; Bryobia praetiosa; Panonychus spp. ;Metatetranychus spp. , and Oligonychus spp. the family Tetranychidae, including Carpoglyphus spp; the family Penthaleidae spp; the family Demodicidae, including Demodex spp; the family Trombicidea, including Trombicula spp; the family Macronyssidae, including Ornothonyssus spp;The family Pyemotidae, including Pyemotes tritici; Tyrophagus putrescentiae; Acaridae, including Acarus siro; Araneidae, including Latrodectus mactans, Tegenaria agrestis, Chiracanthium sp., Lycosa sp. Achaearanea tepidariorum, and Loxosceles reclusa, root-knot nematodes Meloidogyne spp.; cyst nematodes Globodera spp.; Heterodera spp. ; Seed gall nematodes Anguina spp.; Stem and foliar nematodes Aphelenchoides spp.; Sting nematodes Belonolaimus spp.; Pine nematodes Bursaphelenchus spp.; Ring nematodes Criconema spp.; Criconemella spp.; and Criconemoides spp. Stem and bulb nematodes Ditylenchus spp.; ; Owl nematodes Dolichodorus spp. ; Spiral nematodes Heliocotylenchus multicinctus ; Sheath and sheathoid nematodes Hemicycliophora spp. and Hemicriconemoides spp. ; Hirshmanniella spp. ; Lance nematodes Hoploaimus spp. ; False rootknot nematodes, Nacobbus spp.; Needle nematodes, Longidorus spp.; Lesion nematodes, Pratylenchus spp.; Burrowing nematodes, Radopholus spp.; Rhadopholus spp.; Rhodopholus spp.; Reniform nematodes, Rotylenchus spp.; Scutellonema spp. ;Stubby-root nematodes Trichodorus spp.;Paratrichodorus spp.;Stunt nematodes Tylenchorhynchus spp.;Citrus nematodes Tylenchulus spp.;Dagger nematodes Xiphinema spp. and other plant parasitic nematode species, Calotermes flavicollis, Coptotermes spp; Cornitermes cumulans, Cryptotermes spp; Globitermes sulfureus, Heterotermes spp; Leucotermes flavipes, Odontotermes spp., Incisitermes spp; Marginitermes hubbardi, Mastotermes spp, Neocapritermes spp;Neotermes spp, Procornitermes spp, Zootermopsis spp, Reticulitermes spp;Termes natalensis, Blatta spp; Blattella spp; Leucophaea maderae, Panchlora nivea, Periplaneta spp; Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnocelus surinamensis, Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp., Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica, Geophilus spp., Scutigera spp., Blaniulus guttulatus, Julus spp., Narceus spp., Scutigerella immaculatea, Forficula auricularia, Onychiurus spp., Armadillidium vulgare, Oniscus asellus, Porcellio scaber, Damalinia spp., Pediculus spp.; Pthirus pubis, Haematopinus spp.; Linognathus spp.; Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes spp.;Drrissena spp.、Arion ppp.、Bomphalarappppppppppppppppppppppppppppppppptppppppppppppppppppppppppppppppppppttet caaliclla、Scccmea spp.、Ancylostoma duodlle、ceylstllarramlmmeatean lubricomdes、Aacaris rpp.、Brusa malayi、｢rrroririrtirtintintitititititit spp.、Dccrocelfem spp.、Dcctyocaalus flarfa、Dphyllotthriemlisimmmemein rriulosus、Echmococcus multilocularis、Entrobfus vermicularis、Fafmasspp.、5emsssssssThe method according to any one of claims 1 to 3, selected from the group consisting of Heterakis spp., Hymenolepis Nanchung a, Hyostrongulus spp., Loa Loa, Nematodirus spp., Oesophagostomum spp., Opisthorchis spp., Onchocerca volvulus, Ostertagia spp., Paragonimus spp., Schistosomen spp., Strongyloides fuelleborn i, Strongyloides stercoralis, Strongyloides spp., Taenia saginata, Taenia solium, Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella nelsoni, Trichinella pseudopsiralis, Trichostrongulus spp., Trichuris trichiura, Wuchereria bancrofti;

14. 1. A method for determining whether a candidate compound directly binds to a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein, comprising: (a) providing a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein; (b) contacting the complex with a candidate compound; (c) determining the binding affinity of the candidate compound to the complex.

15. The method of claim 14 , wherein the candidate compound is an unlabeled compound.

16. The method of claim 14 , wherein the candidate compound is a labeled compound.

17. The binding affinity is K ｄ and / or K ｉ The method according to any one of claims 14 to 16, wherein the equilibrium binding constant is determined using one or more equilibrium binding constants comprising:

18. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －７ 17. The method of claim 16, wherein the number of sigma-doped ions is equal to or less than M.

19. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －８ 17. The method of claim 16, wherein the number of sigma-doped ions is equal to or less than M.

20. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －９ 17. The method of claim 16, wherein the number of sigma-doped ions is equal to or less than M.

21. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －１０ The method according to any one of claims 14 to 16, wherein the β-

22. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －１１ The method according to any one of claims 14 to 16, wherein the β-

23. The equilibrium dissociation constant (K ｄ ) is about 3 x 10 －１２ The method according to any one of claims 14 to 16, wherein the β-

24. The method of claim 16 , wherein the candidate compound is labeled with a radioisotope.

25. 25. The method of claim 24, wherein the candidate compound is labeled with a fluorophore.

26. 26. The method of claim 24 or 25, wherein the labeled compound is afidopiropen.

27. 1. A method for classifying candidate compounds, comprising: (a) a binding assay step of contacting a complex consisting of an insect transient receptor potential channel Nanchung protein and an insect transient receptor potential channel Water witch protein with the candidate compound and determining the binding affinity of the candidate compound to the complex; (b) identifying a mechanism of action class of the candidate compound.

28. The binding assay step comprises: (1) providing the complex; (2) contacting the complex with a labeled or unlabeled candidate compound; (3) measuring the amount of the candidate compound bound to the complex; (4) The equilibrium dissociation constant (K ｄ ), wherein the K ｄ Hereinafter, (I) (a) Maximum coupling (B ｍａｘ ) and (b) the concentration of the candidate compound required to reach 50% of maximum binding, (II) the association of the candidate compound (K ｏｎ ) and dissociation (K ｏｆｆ ) rate measurement (where K ｄ =K ｏｆｆ / K ｏｎ (III) fluorescence polarization assay; (IV) surface plasmon resonance; (V) isothermal calorimetry; or (VI) thermophoresis. (5) comparing the equilibrium dissociation constant of the candidate compound with the equilibrium dissociation constants of other candidate compounds; and (6) The equilibrium dissociation constant of the candidate compound is calculated based on the equilibrium inhibition constant of a second candidate compound (K ｉ 28. The method of claim 27, comprising the step of comparing the

29. The binding assay step comprises: (1) providing the complex; (2) contacting the complex with the candidate compound and one labeled compound having high binding affinity for the complex; (3) increasing the concentration of unlabeled candidate compound available for contact with the complex until binding of labeled compound to the complex is close to 0%; (4) determining the concentration of the candidate compound required to reduce the amount of bound labeled compound by 50%; (5) The equilibrium inhibition constant (K ｉ ), (6) The equilibrium inhibition constant (K ｉ ) with the inhibition constants of other candidate compounds; (7) The equilibrium inhibition constant of the candidate compound is calculated by multiplying the equilibrium dissociation constant (K ｄ ), and (8) identifying one or more candidate compounds having a high affinity for the complex.

30. 30. The method of claim 14 or 27, wherein binding of the candidate compound to the complex interferes with insect behavior.

31. 30. The method of claim 14 or 27, wherein binding of the candidate compound to the complex kills the insect.

32. 30. The method of claim 14 or 27, wherein binding of the candidate compound to the complex repels insects.

33. 28. The method of claim 14 or 27, wherein the candidate compound is selected from the group consisting of small organic molecules, small inorganic molecules, polysaccharides, peptides, proteins, nucleic acids, extracts made from biological materials, and any combination thereof.

34. 28. The method of claim 14 or 27, wherein the insect is an agricultural / horticultural pest, a disease vector, or a parasite.

35. The insect is an insect from the order Lepidoptera, an insect from the order Coleoptera, an insect from the order Diptera, an insect from the order Thysanoptera, an insect from the order Hemiptera, an insect from the order Hymenoptera, an insect from the order Orthoptera, a pest from the class Arachnida, a pest from the phylum Nematoda, an insect from the order Isoptera, an insect from the order Blattaria, an insect from the order Siphonoptera, 28. The method of claim 14 or 27, wherein the insect is selected from the group consisting of insects from the order Thysanura, pests from the class Chilopoda, pests from the class Diplopoda, pests from the class Symphyla, insects from the order Dermaptera, insects from the order Collembola, pests from the order Isopoda, pests from the phylum Mollusca, pests from the class Bivalvia, pests from the class Gastropoda, and pests from the class Helminthi. [36] The aforementioned insects, Achroia grisella, Acleris spp. argiellacea、Aerrodicus dispersus、Alsophila pômetaria、Ambelophaga rubiginosa、Ammelois transitella、Anacampsis sarcitella、Anagasta Kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia (=Thermesia) spp.; Apamea spp., Aproaerema modicella, Archips spp.; Argyrestia conjugella, Argyroploce spp., Argyrotaenia spp.; Athetis mindara, Austriasca viridigrisea, Autographa gamma, Autographa nigrisigna, Barathra brassicae, Bedelia spp., Bonagota salubricola, Borbo cinnara, Bucculatrix thurberiella, Bupalus piniarius, Busseola spp., Cacoecia spp.; Cactoblastis cactorum, Cadra cautiona, Calingo braziliensis, Caloptilis theivora, Capua reticulana、Carpōsina spp.;Cephus spp、Chaetocnema aridula、Cheimatobia brumata、Chilo spp.;Choreutis pariana、Choristoneura spp;Cyrphis unipuncta, Clysia ambiguella, Cnaphalocerus spp., Cnaphalocrocis medinalis, Cnephasia spp., Cochylis hospes, Coleophora spp., Colias eurytheme, Conopomorpha spp., Conotrachelus spp., Copitarsia spp., Corcyra cephalonica, Crambus caliginosellus, Crambus Teterrellus, Crocidosema (=Epinotia) aporema, Cydalima (=Diaphania) perspectalis, Cydia (=Carpocapsa) spp.; Dalaca noctuides, Datana integrrima, Dasychira pinicola, Dendrolimus spp.; Desmia funeralis, Diaphania spp.; Diatraea grandiosella, Diatraea saccharalis, Diphthera Festiva, Earias spp.; Ecdytolopha aurantianu, Egira (= Xylomyges) curialis, Elasmopalpus lignosellus, Eldana saccharina, Endopiza vitiana, Ennomos subsignaria, Eoreuma loftini, Ephestia spp.; Epinotia aporema, Epiphyas postvittana, Eranis tiliaria, Erionota thrax, Etiella spp.、Eulia spp.、Eupoecilia ambiguella、Euproctis chrysorrhoea、Euxoa spp.、Evetria bouliana、Farona albilinea、Feltia spp.;Galleria m---;;;racillaria spp.;Grapholita spp.;Halysidota spp.;Harrisina americana;Hedylepta spp.;Helicoverpa spp;Heliothis spp.; Hellula spp.; Helocoverpa gelotopoeon, Hemileuca oliviae, Herpetogramma licarsisalis, Hibernia defoliaria, Hofmannophila pseudospretella, Homoeosoma electellum, Homona magnanchungima, Hypena scabra, Hyphantria cunea, Hyponomeuta padella, Hyponomeuta Malinella, Kakivoria flavofasciata, Keiferia lycopersicella, Lambdina ficellaria ficellaria, Lambdina ficellaria lugubrosa, Lamprosema indicata, Laspeyresia molesta, Leguminivora glycinivorell, Lerodea euphala, Leucinodes orbonalis, Leucoma salicis, Leucoptera Ap;Leemmmmvora ]ymmgvorellstitinetineneeeeeeeereierereieieeieereereeieiiiteeeneeeererererer botrana、Lootcampa spp;;tagrotis albicosta;tostege spp;;ymmantria spp;;yonetia clerkella;yonetia #renfoliell;Mesoooma ;Mmestra spp.;Mastra brassicae, Manduca spp.;Marasmia spp., Marmara spp., Maruca testulalis, Megalopyge lanata, Melanchra picta, Melanitis leda, Mocis spp.;Mocis latipes, Monochroa fragariae, Mythimna separatea, Nemapogon cloacella, Neoleucinodes elegantis, Nepytia spp., Nymphula spp., Oiketicus spp., Omiodes indicata, Omphisa anastomosalis, Operophtera brumata, Orgyia pseudotsugata, Oria spp., Orthaga Thyrisalis, Ostrinia spp.; Oulema oryzae, Paleacrita vernata, Panolis flammea, Parnara spp., Papaipema nebris, Papilio cresphontes, Paramyelois transitella, Paranthrene regalis, Paysandisia archon, Pectinophora spp.; Peridroma saucia, Perileucoptera spp.; Phylera bucephala, Phryganidia california, Phthorimaea spp.; Phyllocnistis citrella, Phyllonorycter spp.; Pieris spp.; Pilocrocis tripunctata, Platypena scabra, Platynota spp.; Platyptilia carduidactyla, Plebejus argus, Plodia interpunctella, Plusia spp, Plutella maculipennis, Plutella xylostella, Pontia protodica, Prays spp., Prodenia spp., Proxenus lepigone, Pseudaletia spp.; Pyrausta nubilalis, Rachiplusia nu, Richia albicosta, Rhizobius ventralis, Rhyacionia frustrana, Sabulodes aegrotata, Schizura concinna, Schoenobius spp., Schreckensteinia festaliella, Scirpophaga spp.; Scotia segetum, Sesamia spp., Seudyra subflava, Sitotroga cerealella, Sparganothis pilleriana, Spilonota lechriaspis, S. ocellana, Spodoptera (= Lampygma) spp.;Stig. licus、Thaummatopoｪea pityocampa、Thaumatotibia(==｡yyptophlebia) leucotreta、Thaummatopoea pityocampa、Thecla spp.、Theresima Ampelophaga, Thyrintheina spp., Tildenia inconspicuella, Tinea spp.; Tineola bisselliella, Tortrix spp.; Trichophaga tapetzella, Trichoplusia spp.; Tuta (= Scrobipalpula) absolute, Udea spp.; Virachola spp., Yponomeuta padella, and Zeiraphera canadensis, Acalymma vittatum, Acanthoscehdes obtectus, Adoretus spp., Agelastica alni, Agrilus spp.; Agriotes spp.; Alphitobius diaperinus, Amphimallus solstitialis, Anisandrus dispar, Anisoplia austriaca, Anobium punctatum, Anomala corpulenta, Anomala rufocuprea, Anoplophora spp.; Anthonomus spp.; Anthrenus spp., Aphthona euphoridae, Apion spp., Apogonia spp., Athous haemorrhoidalis, Atomaria spp.; Attagenus spp., Aulacophora femoralis, Blastophagus piniperda, Blitophaga undata, Bruchidius obtectus, Bruchus spp.;Byctiscus betulae, Callidiellum rufipenne, Callopistria floridensis, Callosobruchus chinensis, Cameraria ohridella, Cassida nebulosa, Cerotoma trifurcata, Cetonia aurata, Ceuthorhynchus spp.; Chaetocnema tibialis, Cleonus mendicus, Conoderus spp.; Conotrachellus Water lily, Cosmopolites spp., Costelytra zealandica, Crioceris asparagus, Cryptolestes ferrugineus, Cryptohynchus lapathy, Ctenicera spp.; Curculio spp., Cylindrocopturus spp., Cyclocephala spp., Dactylispa balyi, Dectes texanus, Dermestes spp., Diabrotica spp.; Diaprepes abbreviations, Dichocrocis spp., Dicladispa armigera, Diloboderus abderus, Diocalandra frumenti (Diocalandra stigmaticollis), Enaphalodes rufulus, Epilachna spp.; Epitrix spp.; Eutheola humilis, Eutinobothrus brasiliensis, Faustinus cubae, Gibbium Psylloides, Gnathocerus cornutus, Hellula undalis, Heteronychus arator, Hylamorpha elegans, Hylobius abietis, Hylotrupes bajulus, Hypera spp.;Hypomeces squamosus, Hypothenemus spp., Ips typographus, Lachnosterna consanguinea, Lasioderma serricorne, Latheticus oryzae, Lathridius spp., Lema spp.; Leptinotarsa ​​spp.; Leptispa pygmaea, Limonius californicus, Lissorhoptrus oryzophilus, Lixus spp., Luperodes spp., Lyctus spp.; Liogenys fuscus , Macrodactylus spp.; Maladera matrida, Megaplatypus mutates, Megascelis spp., Melanotus communis, Meligethes spp.; Melolontha spp.; Metamasius hemipterus, Microtheca spp., Migdolus spp., Monochamus spp.; Naupactus Xanthographus, Niptus hololeucus, Oberia brevis, Oemona hirta, Oryctes rhinoceros, Oryzaephilus surinamensis, Oryzaphagus oryzae, Otiorhynchus sulcatus, Otiorhynchus ovatus, Otiorhynchus sulcatus, Oulema melanopus, Oulema oryzae, Oxycetonia jucunda, Phaedon spp;PForacantha recurva、F･y-lobius pyf、F･y-lpertha horticola、F･mlophaga @;EyAAreta s@;Phyllophertha horticola, Popillia japonica, Premnotrypes spp., Psacothea hilaris, Psylliodes chrysocephala, Prostephanus truncates, Psylliodes spp., Ptinus spp., Pulga saltona, Rhizopertha dominica, Rhynchophorus spp.; Saperda candida, Scolytus schevyrewi, Scyphophorus Acupunctatus, Sitona lineatus, Sitophilus spp.; Sphenophorus spp.; Stegobium paniceum, Sternechus spp.; Strophomorphus ctenotus, Symphyletes spp., Tanymecus spp., Tenebrio molitor, Tenebrioides mauretanicus, Tribolium spp.; Trogoderma spp., Tychius spp., Xylotrechus spp.; and Zabrus spp., Aedes spp.; Anastrepha ludens, Anopheles spp.; Bactrocera invadens, Bibio hortulanus, Calliphora erythrocephala, Calliphora vicina, Ceratitis capitata, Chrysomyia spp.; Chrysops atlanticus, Chrysops discalis, Chrysops silacea, Cochliomyia spp.; Contarinia spp.; Cordylobia anthropophaga, Culex spp.; Culicoides furens, Culiseta inornata, Culiseta melanura, Cuterebra spp., Dacus cucurbitae, Dacus oleae, Dasineura brassicae, Dasineura oxycoccana, Delia spp.;Dermatobia hominis, Drosophila spp., Fannia spp.; Gastraphilus spp.; Geomyza tipunctata, Glossina spp.; Haematobia irritans, Haplodiplosis equestris, Hippelates spp., Hylemyia spp.; Hypoderma spp.; Hippobosca spp., Hydrellia philippina, Leptoconops Torrens, Liriomyza spp.; Lucilia spp.; Lycoria pectoralis, Mansonia titillanus, Mayetiola spp.; Musca spp.; Muscina stabulans, Oestrus spp.; Opomyza florum, Oscinella spp.; Orseolia oryzae, Pegomya hysocyami, Phlebotomus argentipes, Phorbia spp.; ;Phytomyza gymnostoma, Prosimulium mixtum, Psila rosae, Psorophora columbiae, Psorophora discolor, Rhagoletis spp.;Rivellia quadrifasciata, Sarcophaga spp.;Simulium vittatum, Sitodiplosis mosellana, Stomoxys spp.;Tabanus spp.;Tannia spp.;Thecodiplosis japonensis, Tipula oleracea, Tipula paludosa, and Wohlfahrtia spp., Baliothrips biformis, Dichromothrips corbetti, Dichromothrips ssp., Echinothrips americanus, Enneothrips flavens, Frankliniella spp.; Heliothrips spp., Hercinothrips femoralis, Kakothrips spp.、MAoce.stttetteteteteeteteterer crentatus、Scirtothrips spp.;Stenchaetothrips spp、Taeniothrips cardamoni、Taeniothrips inconsequens、Thrips spp.、Aciżziaナメックック、ファータッック31.;Acyrrhosin 31.;Adelphicus laricus, Adelphicus tsugae, Adelphicus 31.;Aeneolamia spp., Agonoscena spp., Aulacorthum solani, Aleuracanthus woglumi, Aleurodes spp., Aleurodicus disperses, Aleurolobus barodensis, Aleurothrixus spp., Amrasca spp., Anasa tristis, Antestiopsis spp., Anuraphis cardui, Aonidella spp., Aphanostigma ;;;;;;;;Arboridia apicalis;Arilus critatus;Aspidiella spp.;Aspidiotus spp.;Atanus spp.、Aulacaspis yasumatsui、Aulacorthum solani、Bactericera cockerelli (Paratrioza cockerelli), Bemisia spp.;Blissus tail spp.; Brachycolus spp., Brachycorynella asparagus, Brevicoryne brassicae, Cacopsylla spp.; Calligypona marginata, Calocoris spp., Campylomma livida, Capitophorus horni, Carneocephala fulgida, Cavelerius spp., Ceraplastes spp., Ceratovacuna lanigera, Ceroplastes cerifferus、Cerosipha gossypii、Chaetosiphon fragaefolii、Chionaspis tegalensis、Chlorita onukii、Chromaphis juglandicola、;yoomｾaaffaoorionionionionmmimmmmmimmmmmem spp.; Coccomytilus halli; Coccos spp.Corythucha arcuata, Creontiades dilutes, Cryptomyzus ribis, Chrysomphalus aonidum, Cryptomyzus ribis, Ctenarytaina spatulata, Cyrtopeltis notatus, Dalbulus spp., Dasinus piperis, Dialeurodes spp.; Dalbulus maidis, Diaphorina spp.; Diaspis spp.; Dichelops furcatus, Diconocoris hewetti, Doralis spp., Dreyfusia nordmannianae, Dreyfusia piceae, Drosicha spp., Dysaphis spp.; Dysaulacorthum pseudosolani, Dysdercus spp.; Dysmicoccus spp., Edessa spp., Geocoris spp., Empoasca spp.; Epidiaspis leperii, Eriosoma spp.; Erythroneura spp., Eurygaster spp.; Euscelis bilobatus, Euschistus spp.; Fiorinia theae, Geococcus coffeae, Glycaspis brimblecombei, Halyomorpha spp.; Heliopeltis spp., Homalodisca vitripennis (= H. coagulata), Horcias nobilellus, Hyalopterus pruni, Hyperomyzus Lactucae, Icerya spp.; Idiocerus spp., Idioscopus spp., Laodelphax striatellus, Lecanium spp., Lecanoideus floccissimus, Lepidosaphes spp.; Leptocorisa spp., Leptoglossus phyllopus, Lipaphis erysimi, Lygus spp.;Maconellicoccus hirsutus, Marchalina hellenica, Macropes excavatus, Macrosiphum spp.; Macrosteles quadrilineatus, Mahanarva fimbriolata, Megacopta cribraria, Megaura viciae, Melanaphis pyrarius, Melanaphis sacchari, Melanocallis (= Tinocallis) caryaefoliae, Metcafiella spp., Metopolophium dirhodum, Monellia costalis, Monelliopsis pecanis, Myzocallis corylii, Murgantia spp., Myzus spp.; Nasonovia ribis-nigri, Neotoxoptera formosana, Neomegalotomus spp., Nephotettix spp.; Nezahara spp.; Nilaparvata lugens, Nysius huttoni, Oebalus spp.; Oncometopia spp., Orthezia praelonga, Oxycaraenus hyalinipennis, Parabemisia myricae, Parlatoria spp., Parthenolecanium spp.; Pemphigus spp.; Peregrinus maidis, Perkinsiella saccharicida, Phenococcus spp.; Phloeomyzus passerini, Phorodon humuli, Phylloxera spp., Piesma quadrata, Piezodorus spp.; Pinnaspis aspidistrae, Planococcus spp.; Prosapia bicincta, Protopulvinaria pyriformis, Psallus seriatus, Pseudocysta persea, Pseudoulacaspis pentagona, Pseudococcos spp;Pteromalus spp., Pulvinaria amygdali, Pyrilla spp., Quadraspidiotus spp.; Quesada gigas, Rastrocococcus spp., Reduvius senilis, Rhizoecus americanus, Rhodnius spp., Rhopalomyzus ascalonicus, Rhopalosiphum spp.; Sagatodes spp., Sahlbergella singularis, Saissetia spp., Sappaphis mala, Sappaphis mali, Scaptocoris spp., Scaphoides titanus, Schizaphis graminum, Schizoneura lanuginosa, Scotinophora spp., Selenaspidus articulates, Sitobion avenae, Sogata spp., Sogatella furcifera, Solubea insularis, Spissistilus Festinus (= Stictocephala festina ); Stephanitis nashi, Stephanitis pyrioides, Stephanitis takeyai, Tenalaphara malayensis, Tetraleurodes perseae, Therioaphis maculate, Thyanta spp.; Tibraca spp.; Thomaspis spp.; Toxoptera spp.; Trialeurodes spp.; Triatoma spp., Trioza spp., Typhlocyba spp., Unaspis spp.; and Viteus vitifolii, Acanthomyops interjectus, Athalia rosae, Atta spp., Bombus spp., Brachymyrmex spp., Camponotus spp.; Cardiocondyla nuda, Chalibion ​​sp., Crematogaster spp., Dasymutilla occidentalis, Diprion spp., Dolichovespula maculata, Dorymyrmex spp., Dryocosmus kuriphilus, Formica spp., Hoplocampa spp.; Iridomyrmex humilis, Lasius spp., Linepithema humile, Liometopum spp., Leptocybe invasa, Monomorium spp., Monomorium, Nylandria fulva, Pachycondyla chinensis, Paratrechina longicornis, Paravespula spp; Pheidole spp; Pogonomyrmex spp, Polistes rubiginosa, Prenolepis impairs, Pseudomyrmex gracilis, Schelipron spp, Sirex cyaneus, Solenopsis spp, Sphecius speciosus, Sphec spp, Tapinoma spp; Tetramorium spp, Vespa spp.;Vespula spp.;Wasmannia auropunctata, Xylocopa sp., Acheta domesticus, Calliptamus italicus, Chortoicetes terminifera, Ceutophilus spp., Diastrammena asynamora, Dociostaurus maroccanus, Gryllotalpa spp.;;;y-;us spp;;;;yyyphus daganensis;;raussaria angulifera;Locusta spp;Melano;us spp;Nomadacris septemfasciata;Oedaleus senegalensis、Scapteriscus spp、Schistocerca spp、Stemópelmatus spp、Tachycines asynamorus、and Zonozerus Variegatus; Argasidae, Ixodidae and Sarcoptidae families (Acari), Amblyomma spp., Argas spp., Boophilus spp., Dermacentor spp., Hyalomma spp., Ixodes spp., Rhipicephalus sanguineus, Ornithodorus spp., Ornithonyssus bacoti, Otobius megnini, Dermanyssus gallinae, Psoroptes spp. vitis、E･･｡nmmerus ･yg、Pymlcoy｡truta oleivora;･････tes ribis･･････es spp･････････････ spp.、P･ytonemus pallidus and Polyphagotarsonemus latus, Stenotarsonemus spp. including Stenotarsonemus spinki, Tarsonemidae family; Brevipalpus spp. including Tenuipalpidae family; Eotetranychus spp., Eutetranychus spp., Oligonychus spp., Petrobia latens, Tetranychus spp.; Bryobia praetiosa; Panonychus spp.;Metatetranychus spp., and Oligonychus spp. family Tetranychidae including; Vasates lycopersici; Raoiella indica; family Carpoglyphidae including Carpoglyphus spp; Penthaleidae spp; family Demodicidae including Demodex spp; family Trombicidea including Trombicula spp; family Macronyssidae including Ornothonyssus spp; family Pyemotidae including Pyemotes tritici; Tyrophagus putrescentiae; Acarus siro; Araneidae including Latrodectus mactans, Tegenaria agrestis, Chiracanthium sp, Lycosa sp Achaearanea tepidariorum, and Loxosceles reclusa, root-knot nematodes Meloidogyne spp.; cyst nematodes Globodera spp.; Heterodera spp.; seed gall nematodes Anguina spp. ; stem and foliar nematodes Aphelenchoides spp.; sting nematodes Belonolaimus spp.; pine nematodes Bursaphelenchus spp.; ring nematodes Criconema spp.; Criconemella spp.; and Criconemoides spp.; stem and bulb nematodes Ditylenchus spp. ; Owl nematodes Dolichodorus spp.; Spiral nematodes Heliocotylenchus multicinctus;Sheath and sheathoid nematodes Hemicliophora spp. and Hemicriconemoides spp.; Hirshmanniella spp.; Lance nematodes Hoploaimus spp.; False rootknot nematodes Nacobbus spp.; Needle nematodes Longidorus spp.; Lesion nematodes Pratylenchus spp. ; Burrowing nematodes, Radophorus spp.; Rhadopholus spp.; Rhodopholus spp.; Reniform nematodes, Rotylenchus spp.; Scutellonema spp.; Stubby-root nematodes, Trichodorus spp.; Paratrichodorus spp.; Stunt nematodes, Tylenchorhynchus spp. ; Citrus nematodes, Tylenchulus spp.; Dagger nematodes, Xiphinema spp.; and other plant parasitic nematode species, Calotermes flavicollis, Coptotermes spp; Cornitermes cumulans, Cryptotermes spp; Globitermes sulfureus, Heterotermes spp; Leucotermes flavipes, Odontotermes spp. , Incitermes spp; Marginitermes hubbardi, Mastotermes spp, Neocapritermes spp; Neotermes spp, Procornitermes spp, Zootermopsis spp, Reticulitermes spp. ;Termes natalensis, Blatta spp;Blattella spp;Leucophaea maderae, Panchlora nivea, Periplaneta spp; Supella longipalpa, Parcoblatta pennsylvanica, Eurycotis floridana, Pycnocelus surinamensis, Cediopsylla simples, Ceratophyllus spp., Ctenocephalides spp., Xenopsylla cheopis, Pulex irritans, Trichodectes canis, Tunga penetrans, and Nosopsyllus fasciatus, Lepisma saccharina, Ctenolepisma urbana, and Thermobia domestica, Geophilus spp., Scutigera spp., Blaniulus guttulatus, Julus spp., Narceus spp., Scutigerella immaculatea, Forficula auricularia, Onychiurus spp., Armadillidium vulgare, Oniscus asellus, Porcellio scaber, Damalinia spp., Pediculus spp.; Pthirus pubis, Haematopinus spp.; Linognathus spp.; Bovicola bovis, Menopon gallinae, Menacanthus stramineus and Solenopotes capillatus, Trichodectes spp.;Drrissena spp.、Arion ppp.、Bomphalarappppppppppppppppppppppppppppppppptppppppppppppppppppppppppppppppppppttet caaliclla、Scccmea spp.、Ancylostoma duodlle、ceylstllarramlmmeatean lubricoides, Ascaris spp., Brugia malayi, Brugia timori, Bunostomum spp., Chabertia spp., Clonorchis spp., Cooperia spp., Dicrocoelium spp., Dictyocaulus filaria, Diphyllobothrium latum, Dracunculus; The method according to claim 14 or 27, selected from the group consisting of *Medinensis*, *Echinococcus granulosus*, *Echinococcus multilocularis*, *Enterobius vermicularis*, *Fasciola spp.*, *Haemonchus spp.*; *Heterakis spp.*, *Hymenolepis nanchungae*, *Hyostrongylus spp.*, *Loaloa*, *Nematodirus spp.*, *Oesophagostomum spp.*, *Opisthorchis spp.*, *Onchocerca volvulus*, *Ostertagia spp.*, *Paragonimus spp.*, *Schistosomen spp.*, *Strongyloides fuellebornii*, *Strongyloides stercoralis*, *Strongyloides spp.*, *Taenia saginata*, *Taenia solium*, *Trichinella spiralis*, *Trichinella nativa*, *Trichinella britovi*, *Trichinella nelsoni*, *Trichinella pseudopsiralis*, *Trichostrongylus spp.*, *Trichuris trichiura*, *Wuchereria bancrofti*.

Images