Reagent for detecting methyl salicylate, methyl salicylate sensor, method for sensing methyl salicylate using the same, and method for detecting pathogenic fungus infection of plant

Abstract

To provide a reagent and a methyl salicylate sensor for detecting methyl salicylate which is a plant hormone released when a plant is infected with a pathogenic fungus in the cultivation of the plant including a field crop, to provide a method for sensing methyl salicylate using the same, and to provide a method for detecting the pathogenic fungus infection of the plant.SOLUTION: A reagent for detecting methyl salicylate, comprising a terbium compound and an amine. A methyl salicylate sensor includes a methyl salicylate capture part having the reagent, and a detection part for detecting that methyl salicylate is captured by the capture part.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure relates to a reagent containing a terbium compound and an amine for detecting methyl salicylate, a plant hormone released when plants are infected with pathogenic bacteria, a methyl salicylate sensor, a method for sensing methyl salicylate using the same, and a method for detecting pathogenic bacterial infection in plants. [Background technology]

[0002] It is known that when plants are infected with pathogenic fungi, they synthesize and release plant hormones, which act as signal substances, informing surrounding plants of the pathogenic fungal infection and activating their defense mechanisms in advance. By quickly recognizing these signal substances released by plants, it is possible to detect damage caused by pests and diseases.

[0003] As a method for detecting pest damage to target plants, for example, Patent Document 1 discloses a method in which a monitor plant having a photoprotein gene is placed near the target plant and the monitor plant detects volatile substances released by the target plant in response to stress and emits light. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2019 / 082942 Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present disclosure is to provide a reagent for detecting methyl salicylate, a plant hormone released when plants, including agricultural crops, are infected with pathogens during cultivation, a methyl salicylate sensor, and a methyl salicylate sensing method using the same, thereby providing a method for detecting pathogenic bacterial infection in plants. [Means for solving the problem]

[0006] The present inventors have conducted extensive research to solve the above-mentioned problems, and as a result have found that using a reagent containing a terbium compound and an amine as a receptor for methyl salicylate promotes the reaction between the terbium compound and methyl salicylate, increasing the fluorescence emitted from the complex produced by the reaction, and enabling the detection of plant pathogen infection, thereby completing the present disclosure.

[0007] One aspect of the present embodiment relates to a reagent for detecting methyl salicylate, including a terbium compound and an amine. Another aspect of the present embodiment relates to a methyl salicylate sensor for detecting methyl salicylate, including a capture unit for methyl salicylate having the reagent and a detection unit for detecting capture of methyl salicylate by the capture unit.

[0008] Furthermore, one aspect of this embodiment is a method for sensing methyl salicylate, in which methyl salicylate is detected using the reagent, comprising the steps of: (i) reacting a terbium compound with methyl salicylate to form a complex; (ii) exposing the complex to excitation light; (iii) detecting the fluorescence emitted by the complex The present invention relates to a method for sensing methyl salicylate, comprising:

[0009] Another aspect of this embodiment relates to a method for detecting pathogenic fungal infection of a plant by placing the reagent near the plant and confirming fluorescence emitted from a complex formed upon reaction of a terbium compound with methyl salicylate. [Effects of the Invention]

[0010] According to the present disclosure, by incorporating a terbium compound and an amine as a receptor for methyl salicylate in a reagent for detecting methyl salicylate, the intensity of fluorescence emission from the complex formed by the reaction of the terbium compound with methyl salicylate, a volatile plant hormone released when a plant is infected with a pathogenic fungus, is increased, making it possible to detect infection of the plant by a pathogenic fungus. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 shows the fluorescence spectrum curve obtained in the present disclosure and the fluorescence spectrum curve obtained in Comparative Example 1. [Figure 2] FIG. 2 shows the fluorescence spectral curves obtained in this disclosure. [Figure 3] FIG. 3 shows the fluorescence spectral curves obtained in this disclosure. [Figure 4] FIG. 4 shows the fluorescence spectral curves obtained in this disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0012] Hereinafter, embodiments for carrying out the present disclosure will be described with reference to the drawings, etc. However, the embodiments described below are limited to technically preferable aspects for carrying out the present disclosure, but the scope of the invention is not limited to the following.

[0013] [1] Reagent for detecting methyl salicylate One embodiment of the present disclosure is a reagent for detecting methyl salicylate, comprising a terbium compound and an amine as a receptor for methyl salicylate. In this disclosure, the term "reagent" is defined as a chemical substance used to detect or quantify a substance by chemical methods, to experiment with the synthesis of a substance, or to measure a physical property.

[0014] Furthermore, the reagent preferably contains, in addition to the terbium compound and the amine, a non-volatile ionic liquid that effectively captures methyl salicylate. When the reagent contains an ionic liquid, the terbium compound and the amine are present in a dissolved state in the non-volatile ionic liquid, but some may also precipitate. The terbium compound can function as a receptor for methyl salicylate whether dissolved in the non-volatile ionic liquid or precipitated.

[0015] <Receptor for methyl salicylate: terbium compounds> In the present disclosure, terbium compounds that can be used to recognize methyl salicylate include, but are not limited to, terbium compounds represented by the following general formula (1): LncTbXaYb (1)

[0016] In the formula, Tb is terbium, and Ln is at least one rare earth element other than terbium, and specific examples include scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium.

[0017] In the formula, X and Y each independently represent RCOO - R is at least one monovalent anion selected from the group consisting of a halogen atom, a halogen atom, a fluorine atom, a methyl group, an ethyl group, a propyl group, a 2-methylpropyl group, a trifluoromethyl group, a tert-butyl group, or an isobutyl group. -Specific examples thereof preferably include at least one selected from the group consisting of acetate ion, trifluoroacetate ion, propionate ion, butyrate ion, isobutyrate ion, pivalate ion, 2-methylbutyrate ion, benzoate ion, chloride ion, and nitrate ion. Specific examples of the halide ion include fluoride ion, chloride ion, bromide ion, iodide ion, etc., and chloride ion is preferred. X and Y may each be two or more kinds.

[0018] In the formula, a, b, and c are numbers that satisfy 0 < a, 0 ≤ b, 0 ≤ c ≤ 0.05, and a + b = 3 + 3×c, respectively.

[0019] Among the compounds represented by the above general formula (1), specific examples of the terbium compound with b = c = 0 include terbium acetate, terbium propionate, terbium butyrate, terbium isobutyrate, terbium pivalate, terbium benzoate, terbium nitrate, terbium chloride, etc.

[0020] Among the compounds represented by the above general formula (1), specific examples of the terbium compound with b ≠ 0 and c = 0 include terbium acetate-pivalate complex salt, terbium 2-methylbutyrate-pivalate complex salt, terbium trifluoroacetate-pivalate complex salt, terbium acetate-terbium 2-methylbutyrate complex salt, etc. For example, terbium acetate-pivalate complex salt (Tb(CH3COO) a (t-C4H9COO) b ) can be synthesized by completely dissolving terbium acetate and terbium pivalate in methanol and subjecting them to a heating reaction. Also, by changing the mixing ratio of the terbium salts during synthesis, the ratio of the counter ions (X, Y) of the complex salt can be changed.

[0021] Among the compounds represented by the above general formula (1), specific examples of the terbium compound with b ≠ 0 and c ≠ 0 include Ce 0.02 Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Eu 0.02Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Sm 0.02 Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Dy 0.02 Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Gd 0.02 Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Yb 0.02 Tb(CH3COO) 0.81 (t-C4H9COO) 2.25 、Ce 0.01 Tb(CH3COO) 0.78 (t-C4H9COO) 2.25 、Ce 0.015 Tb(CH3COO) 0.795 (t-C4H9COO) 2.25 、Ce 0.025 Tb(CH3COO) 0.825 (t-C4H9COO) 2.25 、Ce 0.04 Tb(CH3COO) 0.87 (t-C4H9COO) 2.25 、Ce 0.05 Tb(CH3COO) 0.9 (t-C4H9COO) 2.25 、Ce 0.06 Tb(CH3COO) 0.93 (t-C4H9COO) 2.25 、Ce 0.02 Tb(CH3COO) 3.06、 Ce 0.04 Tb(CH3COO) 3.12 、Yb 0.15 Tb(CH3COO) 0.795 (t-C4H9COO) 2.25 、Yb 0.025 Tb(CH3COO) 0.825 (t-C4H9COO) 2.25 、Yb 0.04 Tb(CH3COO) 0.87 (t-C4H9COO) 2.25 Even if you are a Ce c Tb(CH3COO) a(t-C4H9COO) b can be synthesized by dissolving terbium acetate, terbium pivalate, and any number of moles of cerium acetate relative to terbium (Tb) in methanol and heating the reaction. By changing the ratio of the two terbium salts (terbium acetate and terbium pivalate) and the molar ratio of cerium acetate during synthesis, the ratio of counter ions (X, Y) and the amount of cerium doped (c) can be changed.

[0022] When doping with a rare earth element (Ln) other than terbium, c is preferably 0.015 to 0.05. When doping with two or more rare earth elements (Ln) other than terbium, the total c is preferably 0.015 to 0.05. When c is within this range, the effect of increasing the fluorescence emission intensity derived from the complex formed by the reaction with methyl salicylate is fully exerted.

[0023] These terbium compounds react with methyl salicylate to form complexes, which enable them to selectively recognize methyl salicylate.

[0024] The resulting methyl salicylate-terbium complex emits fluorescence specific to terbium complexes when excited by UV light. The fluorescence intensity of the terbium compound alone is so low that it cannot be observed even when irradiated with UV light. Furthermore, the terbium compound does not react with or recognize other plant hormones other than methyl salicylate, such as methyl jasmonate, allowing it to selectively recognize methyl salicylate.

[0025] <amine> To promote the reaction between the terbium compound and methyl salicylate, the reagent of the present disclosure includes a basic amine. Amines that can be used in the present disclosure include amines having 6 to 30 carbon atoms, specifically, but not limited to, triethylamine, tripropylamine, tributylamine, triisobutylamine, triamylamine, triisoamylamine, trihexylamine, triheptylamine, tri-n-octylamine, tris(2-ethylhexylamine), tridecylamine, tribenzylamine, triphenylamine, N,N-diethylaniline, etc. These amines include: Below formula As shown in Figure 1, in the reaction between methyl salicylate and a terbium compound, the amine interacts with the hydrogen atom of the phenolic hydroxyl group of methyl salicylate, promoting the reaction between methyl salicylate and the terbium compound, resulting in an increase in fluorescence intensity. The molar ratio of the amine to the terbium compound is preferably 0.5 to 10, more preferably 2 to 7, and particularly preferably 3 to 5. If the molar ratio of the amine to the terbium compound is less than 0.5, the effect of promoting the reaction with methyl salicylate is reduced, and the effect of improving detection sensitivity may not be achieved. If the molar ratio of the amine to the terbium compound is more than 10, the terbium compound may have difficulty forming a complex with methyl salicylate.

[0026] [ka]

[0027] <Non-volatile ionic liquid> To effectively capture methyl salicylate, the reagent of the present disclosure preferably contains a non-volatile ionic liquid, such as an imidazolium salt, a phosphonium salt, a pyridinium salt, an ammonium salt, a piperidinium salt, or a pyrrolidinium salt. Specific examples include 1-ethyl-3-methylimidazolium acetate, 1-butyl-3-methylimidazolium acetate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-butyl-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-butyl-3-methylimidazolium dibutylphosphate, tetrabutylammonium acetate, 1-butylpyridinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-propylpiperidinium bis(fluorosulfonyl)imide, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-butyl-3-methylimidazolium dicyanamide, and the like, but are not limited to these.

[0028] The weight ratio of the ionic liquid to the terbium compound is preferably 2 to 40, more preferably 3 to 30, and particularly preferably 4 to 20. If the weight ratio of the ionic liquid to the terbium compound is less than 2, the effect of improving the detection sensitivity for methyl salicylate may not be obtained, whereas if the weight ratio of the ionic liquid to the terbium compound is more than 40, the terbium compound may have difficulty forming a complex with methyl salicylate.

[0029] The reagent for detecting methyl salicylate of the present disclosure may optionally contain other solvents within the scope that does not impair the effects of the present disclosure, such as, but not limited to, dimethyl sulfoxide, methanol, ethanol, water, N,N-dimethylformamide, tetrahydrofuran, acetone, acetonitrile, and 1,4-dioxane.

[0030] [2] Methyl salicylate sensor One embodiment of the present disclosure relates to a methyl salicylate sensor for detecting methyl salicylate, comprising a capture unit for methyl salicylate having the reagent, and a detection unit for detecting capture of methyl salicylate on the capture unit.

[0031] (1) Capture unit The capture unit of the methyl salicylate sensor of the present disclosure has a terbium compound, which is a receptor that selectively captures methyl salicylate, and a reagent containing an amine that promotes the reaction between the terbium compound and methyl salicylate. In the capture unit, the reagent is preferably contained in a medium.

[0032] <medium> The medium for containing the terbium compound and amine of the present disclosure may be, for example, paper or glass fiber, resin (e.g., polymethyl methacrylate, polyethylene, polypropylene, polyvinyl chloride, polystyrene, nylon resin, polyamide, polycarbonate, polyethylene terephthalate, polybutylene terephthalate, polyphenylene oxide), water-soluble polymer (cellulose-based, agarose, starch-based, sodium alginate, acrylic acid-based, acrylamide-based, polyvinyl alcohol, polyethylene oxide, polyvinylpyrrolidone, etc.), but is not limited to these.

[0033] For example, when paper is used as the medium, a terbium compound and an amine are dissolved in a solvent, the resulting solution is impregnated into paper (e.g., filter paper), and the paper is then dried at room temperature to 60°C to remove the solvent, thereby obtaining a medium containing the terbium compound and the amine. After drying, the solvent is evaporated and removed, but some may remain. The use of a solvent is preferred because it makes it easier to impregnate the medium with the terbium compound and also makes it easier to adjust the concentration of the terbium compound.

[0034] Examples of solvents that can be used to dissolve the terbium compound include, but are not limited to, dimethyl sulfoxide, methanol, ethanol, water, N,N-dimethylformamide, tetrahydrofuran, acetone, acetonitrile, and 1,4-dioxane.

[0035] In order to effectively capture methyl salicylate, it is preferable to add the aforementioned non-volatile ionic liquid to the solvent. When a non-volatile ionic liquid is used, after removing the solvent, the terbium compound is dissolved in the non-volatile ionic liquid, but some may precipitate. The terbium compound can function as a receptor for methyl salicylate whether dissolved in the non-volatile ionic liquid or precipitated.

[0036] The ratio of ionic liquid to solvent can be set appropriately for the medium to be impregnated. If the ratio of ionic liquid to solvent is low, the amount of ionic liquid in the medium after drying will be small, which may reduce the effect of improving detection sensitivity. On the other hand, if the ratio of ionic liquid to solvent is high, the ionic liquid may have the disadvantage of being difficult to impregnate into the medium due to its high viscosity. Therefore, the ratio of ionic liquid to solvent is set appropriately for the medium. For example, when impregnating filter paper, the ratio of ionic liquid to solvent is preferably 5 to 50 wt %, more preferably 10 to 30 wt %.

[0037] (2) Detection unit The detection unit of the methyl salicylate sensor of the present disclosure is configured to optically detect the capture of methyl salicylate by the capture unit. The detection unit may be configured as a separate device rather than being integrated with the capture unit. In one aspect of the present disclosure, the optical detection unit includes an excitation light source (light emitter) and a detection element (fluorescence receiver) to detect the fluorescence emission of a complex formed between a terbium compound and methyl salicylate, and can detect and / or measure the concentration of methyl salicylate based on the observed change in fluorescence intensity.

[0038] In one embodiment of the present disclosure, the detection unit may include a computer that executes a program for processing the detection and / or concentration measurement of methyl salicylate. Such a program may, for example, cause the computer to execute the steps of: (i) receiving a signal from an optical detection element; (ii) analyzing the received signal to determine the presence and / or concentration of methyl salicylate; and (iii) outputting the analysis results.

[0039] In one embodiment of the present disclosure, analyzing the received signal may include, for example, determining the presence and / or concentration of methyl salicylate by comparing the received signal with a predetermined reference value. Also, in one embodiment of the present disclosure, the analysis results may be output, for example, to a display device connected to the sensor or to another device connected via a network.

[0040] In one aspect of the present disclosure, the methyl salicylate sensor of the present disclosure detects methyl salicylate, a plant hormone released when agricultural crops are infected with pathogenic bacteria, and can therefore be used as a sensor for detecting pathogenic bacteria infection in plants, including agricultural crops.

[0041] [3] Methyl salicylate sensing method One embodiment of the present disclosure is a method for sensing methyl salicylate using the above-mentioned reagent or methyl salicylate sensor, utilizing a fluorescence emission phenomenon from a complex obtained by reacting a terbium compound with methyl salicylate, the method comprising: (i) reacting a terbium compound with methyl salicylate to form a complex; (ii) exposing the complex to excitation light; (iii) A method for sensing methyl salicylate, comprising the step of detecting the fluorescence emitted by the complex.

[0042] While terbium compounds alone show almost no fluorescence, the complex formed by the reaction of terbium compounds with methyl salicylate exhibits additional fluorescence. This phenomenon can be utilized to detect methyl salicylate.

[0043] In one embodiment of the present disclosure, an appropriate excitation wavelength within the range of 300 to 400 nm is selected. Furthermore, in one embodiment of the present disclosure, a step of determining the concentration of methyl salicylate by comparing the detected fluorescence intensity with a predetermined reference value may also be performed.

[0044] [4] Methods for detecting plant pathogen infection One embodiment of the present disclosure relates to a method for detecting pathogenic fungal infection in plants by placing the reagent or methyl salicylate sensor near a plant and confirming fluorescence emitted from a complex formed upon reaction between a terbium compound and methyl salicylate.

[0045] Examples of plants that may be monitored include, but are not limited to, cucumber, watermelon, tomato, eggplant, bell pepper, paprika, shishito pepper, melon, Chinese cabbage, cabbage, radish, lettuce, leek, broccoli, onion, garlic, yam, asparagus, carrot, potato, celery, tobacco, rice, and strawberry.

[0046] Diseases that can be detected include, but are not limited to, ring spot, white spot, brown ring spot, downy mildew, wilt, root rot wilt, half-leaf wilt, brown root rot, gray blight, root rot, black spot root rot, white mold, damping-off, brown spot, downy mildew, powdery mildew, gray mold, anthracnose, black spot, sclerotinia, vine blight, spot, late blight, mosaic disease, yellow necrosis, yellow cigar disease, bacterial wilt, soft rot, canker, bacterial stem necrosis, bacterial black spot, and bacterial spot. Pathogenic infections that can be detected include, but are not limited to, infections caused by the causative fungi of the above diseases.

[0047] In the context of the present disclosure, when it is said that a sensor is "placed near a plant," examples of the term "nearby" include, but are not limited to, a distance of within 2 m, 1 m, 75 cm, 50 cm, 40 cm, 30 cm, 20 cm, 10 cm, or 5 cm from the plant to be monitored, and an appropriate distance is appropriately selected in consideration of various factors. A person skilled in the art would be able to appropriately determine the location to place the sensor in consideration of various conditions.

[0048] Furthermore, an embodiment of the present disclosure relates to the use of the reagent or methyl salicylate sensor in detecting pathogen infection in plants. Also, an embodiment of the present disclosure relates to the use of a terbium compound and an amine in the manufacture of the reagent or methyl salicylate sensor. [Example]

[0049] The present disclosure will be explained in more detail below by way of examples, but the present disclosure is not limited to these examples.

[0050] (Synthesis Example 1) Cerium (Ce) doped 2% terbium acetate (Tb(CH3COO)3:TbA)-terbium pivalate (Tb(t-C4H9COO)3:TbPv) complex salt [Ce 0.02 Tb(CH3COO) 0.81 (C4H9COO) 2.25 :0.02CeA-0.25TbA-0.75TbPv] 0.2g of terbium acetate tetrahydrate, 0.6797g of terbium pivalate, and 0.0131g of cerium acetate monohydrate were dissolved in 120ml of methanol and heated under reflux for 5 hours. After cooling, the methanol was distilled off and the precipitated white crystals were dried in a vacuum to obtain 0.733g of the desired terbium compound doped with 2% cerium.

[0051] [Fluorescence behavior upon reaction with methyl salicylate] Example 1 0.0437 g of the 2% cerium-doped terbium acetate-terbium pivalate composite salt (0.02CeA-0.25TbA-0.75TbPv) obtained in Synthesis Example 1 was dissolved in 1.6 ml of dimethyl sulfoxide (DMSO), and 0.07 ml of triethylamine (NEt3) (5 times the molar amount relative to terbium) and 0.4 ml of 1-ethyl-1-methylimidazolium acetate (EMImAc) were added. 0.2 ml of the resulting solution was dropped onto a circular filter paper (Φ40 mm) and dried to volatilize the DMSO, yielding a filter paper containing 0.02CeA-0.25TbA-0.75TbPv, NEt3, and EMImAc. The resulting filter paper was sandwiched between two glass plates (40 mm square, 2 mm thick, with a 1 / 4 inch hole in the center) and secured with a clip. A Teflon tube was inserted into the 1 / 4 inch hole, and methyl salicylate at a concentration of 0.01 μg / L was passed through the tube using a Permeator PD-1B-2 (Gastec Corporation) with nitrogen as the carrier gas. After 1 hour of exposure, the filter paper was removed and the fluorescence spectrum was measured at an excitation wavelength of 365 nm. The fluorescence spectrum obtained in Example 1 is shown by the solid line in Figure 1.

[0052] (Comparative Example 1) Except for not using triethylamine (NEt3), the fluorescence spectrum was measured at an excitation wavelength of 365 nm in the same manner as in Example 1. The fluorescence spectrum obtained in Comparative Example 1 is shown by the dashed line in Figure 1.

[0053] 1, the fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum containing triethylamine (Example 1) was approximately 11 times greater than the fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum not containing triethylamine (Comparative Example 1). Thus, it was revealed that the inclusion of triethylamine facilitates sensing of methyl salicylate in the gas phase at a concentration of 0.01 μg / L.

[0054] Example 2 Evaluation was carried out in the same manner as in Example 1, except that 0.0453 g of terbium nitrate hexahydrate was used instead of the 2% cerium-doped terbium acetate-terbium pivalate composite salt (0.02CeA-0.25TbA-0.75TbPv).

[0055] Figure 2 shows the fluorescence spectrum after 1 hour of exposure (solid line) and the fluorescence spectrum before and after exposure (dashed line). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after 1 hour of exposure increased by approximately 33 times compared to the fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum before and after exposure. Thus, it was demonstrated that gas-phase methyl salicylate at a concentration of 0.01 μg / L can be sensed using a reagent containing a terbium compound and an amine.

[0056] Example 3 Evaluation was carried out in the same manner as in Example 2, except that 0.072 ml of triisobutylamine (5 times the molar amount relative to terbium) was used instead of triethylamine.

[0057] Figure 3 shows the fluorescence spectrum after 1 hour of exposure (solid line) and the fluorescence spectrum before and after exposure (dashed line). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after 1 hour of exposure increased to approximately 32 times the fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum before and after exposure. Thus, it was demonstrated that gas-phase methyl salicylate at a concentration of 0.01 μg / L can be sensed using a reagent containing a terbium compound and an amine.

[0058] Example 4 Evaluation was carried out in the same manner as in Example 2, except that 0.13 ml of tris(2-ethylhexyl)amine (three times the molar amount relative to terbium) was used instead of triethylamine.

[0059] Figure 4 shows the fluorescence spectrum after 1 hour of exposure (solid line) and the fluorescence spectrum before and after exposure (dashed line). In the fluorescence spectrum after 1 hour of exposure, the fluorescence intensity at a wavelength of 544 nm increased by approximately 75 times compared to the fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum before and after exposure. Thus, it was demonstrated that gas-phase methyl salicylate at a concentration of 0.01 μg / L can be sensed using a reagent containing a terbium compound and an amine.

[0060] Example 5 Similar to Example 1, the evaluation was performed using 0.0408 g of terbium acetate tetrahydrate instead of the 2% cerium-doped terbium acetate-terbium pivalate complex salt (0.02CeA-0.25TbA-0.75TbPv). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after one hour of exposure increased approximately eight-fold compared to the unexposed state. Thus, it was demonstrated that the use of a reagent containing a terbium compound and an amine enabled sensing of methyl salicylate in the gas phase at a concentration of 0.01 μg / L.

[0061] Example 6 Similar to Example 1, this evaluation was performed using 0.0373 g of terbium chloride hexahydrate instead of the 2% cerium-doped terbium acetate-terbium pivalate complex salt (0.02CeA-0.25TbA-0.75TbPv). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after one hour of exposure increased approximately 23-fold compared to the unexposed state. Thus, it was demonstrated that the use of a reagent containing a terbium compound and an amine enabled sensing of methyl salicylate in the gas phase at a concentration of 0.01 μg / L.

[0062] Example 7 Similar to Example 1, evaluation was performed using 0.0462 g of terbium pivalate instead of the 2% cerium-doped terbium acetate-terbium pivalate complex salt (0.02CeA-0.25TbA-0.75TbPv). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after one hour of exposure increased approximately 15-fold compared to the unexposed state. Thus, it was demonstrated that gas-phase methyl salicylate at a concentration of 0.01 μg / L can be sensed using a reagent containing a terbium compound and an amine.

[0063] Example 8 The same procedure as in Example 1 was repeated, except that the amount of triethylamine added was changed to 0.042 ml (three times the molar amount relative to terbium). The fluorescence intensity at a wavelength of 544 nm in the fluorescence spectrum after one hour of exposure increased to approximately 13 times the fluorescence intensity before and after exposure. Thus, it was revealed that the inclusion of triethylamine facilitates sensing of methyl salicylate in the gas phase at a concentration of 0.01 μg / L. [Industrial Applicability]

[0064] The methyl salicylate sensor for detecting the plant hormone methyl salicylate according to the present disclosure includes a capture unit having a medium containing a terbium compound and an amine, which efficiently captures methyl salicylate, forms a complex, and emits fluorescence, thereby enabling selective detection of methyl salicylate, a plant hormone released by plants upon pathogen infection. The methyl salicylate sensor can be used to detect pathogen infection in plants, and specifically, can be used as a new sensor for agricultural ICT in greenhouses and other protected horticulture facilities, as a sensor capable of detecting pathogen infection in agricultural crops.

[0065] Although the present disclosure has been described above with reference to the embodiments and examples, the present disclosure is not limited to the above-described embodiments and examples. Various modifications that can be understood by a person skilled in the art can be made to the configuration and details of the present disclosure within the scope of the present disclosure.

[0066] (Appendix 1) A reagent for detecting methyl salicylate, comprising a terbium compound and an amine. (Appendix 2) The reagent according to Appendix 1, wherein the amine is an amine having 6 to 30 carbon atoms. (Appendix 3) 3. The reagent of claim 1 or 2, wherein the amine is at least one selected from the group consisting of triethylamine, tripropylamine, tributylamine, triisobutylamine, triamylamine, triisoamylamine, trihexylamine, triheptylamine, tri-n-octylamine, tris(2-ethylhexylamine), tridecylamine, tribenzylamine, triphenylamine, and N,N-diethylaniline. (Appendix 4) The terbium compound is represented by the following general formula (1): Ln c TbX a Y b (1) (In the formula, Tb is terbium, Ln is at least one rare earth element other than terbium, and X and Y each independently represent RCOO - a, b, and c are each independently 0 or 1; R is at least one monovalent anion selected from the group consisting of a linear or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, and an aryl group having 6 to 14 carbon atoms, each of which is optionally substituted with a halogen atom; <a、0≦b、0≦c≦0.05、及びa+b=3+3×cを満たす数である)The reagent of any preceding claim, wherein the reagent is at least one of the compounds represented by (Appendix 5) The reagent according to any of the preceding appendices, wherein the molar ratio of the amine to the terbium compound is 0.5 to 10. (Appendix 6) 10. The reagent of any preceding clause, wherein the reagent for detecting methyl salicylate further comprises at least one non-volatile ionic liquid selected from the group consisting of 1-ethyl-3-methylimidazolium acetate, 1-butyl-3-methylimidazolium acetate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-butyl-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-butylpyridinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-propylpiperidinium bis(fluorosulfonyl)imide, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, and 1-butyl-3-methylimidazolium methyl dicyanamidate. (Appendix 7) 7. The reagent according to any one of Appendices 4 to 6, wherein X and Y are each independently at least one selected from the group consisting of acetate ion, trifluoroacetate ion, propionate ion, butyrate ion, isobutyrate ion, pivalate ion, 2-methylbutyrate ion, benzoate ion, chloride ion, and nitrate ion. (Appendix 9) The reagent according to any one of Appendices 4 to 8, wherein the compound represented by the general formula (1) is at least one selected from the group consisting of terbium acetate, terbium propionate, terbium butyrate, terbium isobutyrate, terbium pivalate, terbium benzoate, terbium nitrate, and terbium chloride. (Appendix 9) The reagent according to any one of Appendices 4 to 7, wherein c=0. (Appendix 10) 8. The reagent according to any one of Appendices 4 to 7, wherein 0.015≦c≦0.05. (Appendix 11) The reagent according to any one of Appendices 4 to 7 and 10, wherein Ln is at least one selected from the group consisting of scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium. (Appendix 12) The reagent of any preceding clause, further comprising a non-volatile ionic liquid. (Appendix 13) 13. The reagent of claim 12, wherein the non-volatile ionic liquid is at least one selected from the group consisting of imidazolium salts, phosphonium salts, pyridinium salts, ammonium salts, piperidinium salts, and pyrrolidinium salts. (Appendix 14) 13. The reagent of claim 12, wherein the non-volatile ionic liquid is at least one selected from the group consisting of 1-ethyl-3-methylimidazolium acetate, 1-butyl-3-methylimidazolium acetate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-butyl-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-butyl-3-methylimidazolium dibutylphosphate, tetrabutylammonium acetate, 1-butylpyridinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-propylpiperidinium bis(fluorosulfonyl)imide, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, and 1-butyl-3-methylimidazolium dicyanamide. (Appendix 15) 15. The reagent according to any one of Appendices 12 to 14, wherein the ratio of the non-volatile ionic liquid to the terbium compound is 2 to 40 times by weight. (Appendix 16) A methyl salicylate sensor for detecting methyl salicylate, a capture moiety for methyl salicylate having a reagent according to any preceding clause; a detection unit that detects that methyl salicylate has been captured by the capture unit; A methyl salicylate sensor comprising: (Appendix 17) 17. The methyl salicylate sensor of claim 16, wherein the capture portion comprises a medium containing the reagent. (Appendix 18) 18. The methyl salicylate sensor of claims 16 and 17, wherein the medium is paper, glass fiber, resin, or a water-soluble polymer. (Appendix 19) 19. The methyl salicylate sensor according to any one of appendices 16 to 18, wherein the detection unit includes a computer that executes a program for processing the detection and / or concentration measurement of methyl salicylate. (Appendix 20) A method for sensing methyl salicylate, which uses the reagent according to any one of Supplementary Notes 1 to 15 to recognize methyl salicylate, (i) reacting a terbium compound with methyl salicylate to form a complex; (ii) exposing the complex to excitation light; (iii) detecting the fluorescence emitted by the complex A method for sensing methyl salicylate, comprising: (Appendix 21) A method for sensing methyl salicylate, which comprises recognizing methyl salicylate using the methyl salicylate sensor according to any one of Supplementary Notes 16 to 18, (i) reacting a terbium compound with methyl salicylate to form a complex; (ii) exposing the complex to excitation light; (iii) detecting the fluorescence emitted by the complex A method for sensing methyl salicylate, comprising: (Appendix 22) A method for detecting pathogenic fungal infection of a plant, comprising placing a reagent according to any one of Appendices 1 to 15 near a plant and confirming fluorescence emitted from a complex formed upon reaction of a terbium compound with methyl salicylate. (Appendix 23) A method for detecting pathogenic fungal infection of a plant, comprising placing a methyl salicylate sensor according to any one of Appendices 16 to 18 near a plant and confirming fluorescence emitted from a complex formed upon reaction of a terbium compound with methyl salicylate.

Claims

1. A reagent for detecting methyl salicylate, comprising a terbium compound and an amine.

2. 2. The reagent according to claim 1, wherein the amine is an amine having 6 to 30 carbon atoms.

3. 3. The reagent according to claim 1, wherein the amine is at least one selected from the group consisting of triethylamine, tripropylamine, tributylamine, triisobutylamine, triamylamine, triisoamylamine, trihexylamine, triheptylamine, tri-n-octylamine, tris(2-ethylhexylamine), tridecylamine, tribenzylamine, triphenylamine, and N,N-diethylaniline.

4. The terbium compound is represented by the following general formula (1): Ln ｃ TbX ａ 9 ｂ (1) (In the formula, Tb is terbium, Ln is at least one rare earth element other than terbium, and X and Y each independently represent RCOO － , at least one monovalent anion selected from the group consisting of a halide ion and a nitrate ion; R is at least one selected from the group consisting of a linear or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, and an aryl group having 6 to 14 carbon atoms, which are optionally substituted with halogen; and a, b, and c are numbers that satisfy 0<a, 0≦b, 0≦c≦0.05, and a+b=3+3×c, respectively. The reagent according to claim 1 or 2, which is at least one compound represented by the formula:

5. 3. The reagent according to claim 1, wherein the molar ratio of the amine to the terbium compound is 0.5 to 10.

6. 3. The reagent according to claim 1 or 2, wherein the reagent for detecting methyl salicylate further comprises at least one non-volatile ionic liquid selected from the group consisting of 1-ethyl-3-methylimidazolium acetate, 1-butyl-3-methylimidazolium acetate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-butyl-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-butylpyridinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-propylpiperidinium bis(fluorosulfonyl)imide, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, and 1-butyl-3-methylimidazolium methyl dicyanamidate.

7. A methyl salicylate sensor for detecting methyl salicylate, a capture part for methyl salicylate having the reagent according to claim 1 or 2; a detection unit that detects that methyl salicylate has been captured by the capture unit; A methyl salicylate sensor comprising:

8. The methyl salicylate sensor according to claim 7 , wherein the capture portion comprises a medium containing the reagent.

9. A method for sensing methyl salicylate, which uses the reagent according to claim 1 or 2 to recognize methyl salicylate, comprising: (i) reacting a terbium compound with methyl salicylate to form a complex; (ii) exposing the complex to excitation light; (iii) detecting the fluorescence emitted by the complex A method for sensing methyl salicylate, comprising:

10. 3. A method for detecting pathogenic fungal infection of a plant, comprising placing the reagent according to claim 1 or 2 in the vicinity of the plant and confirming fluorescence emitted from a complex formed upon reaction of a terbium compound with methyl salicylate.

Images