Plant stomatal opening regulator

Abstract

The present invention relates to a plant stomatal opening regulator, which contains at least one selected from the group consisting of compounds represented by general formula (1), salts thereof, and solvates thereof.

Description

Technical Field

[0001] The invention relates to a plant stomatal opening regulator. Background Art

[0002] Terrestrial higher plants regulate the intake of carbon dioxide and transpiration required for photosynthesis by regulating the opening of stomata present in the leaf epidermis. For example, it is known that in a water-deficient environment, in order to maintain the moisture in the plant body, the stomata are closed to inhibit transpiration. It is also known that stomata open under the action of light to promote the intake of carbon dioxide required for photosynthesis. Therefore, it is expected to achieve effects such as promoting photosynthesis, promoting growth and improving drought resistance by artificially regulating the opening of stomata.

[0003] As an example of artificial regulation of stomatal opening, a method for guard cells to make their cell membrane H ﹢ -ATPase (AHA2) overexpression method (Patent Document 1, Non-Patent Document 1). According to this method, stomatal opening can be promoted, and photosynthesis rate and plant growth can be promoted accordingly. In addition, a method for improving drought resistance by promoting stomatal closure by overexpressing the H subunit of magnesium chelatase in guard cells has also been reported (Non-Patent Document 2).

[0004] Since these methods are based on gene recombination technology,

[0005] 1. For biological species for which genetic recombination technology has not yet been established, it is necessary to establish and optimize recombination technology each time;

[0006] 2. The production of genetically modified crops (GMO) takes time. For example, it takes about half a year to cultivate the first generation of genetically modified poplars, and in the case of rice, it takes more than half a year to obtain the next generation of GMO seeds;

[0007] 3. Approval is required for every GMO that is cultivated.

[0008] On the other hand, if a substance can regulate stomatal opening by applying it to cultivated plants, it is unnecessary to obtain procedures such as cultivation and approval of GMOs for various plants, and thus it is extremely useful.

[0009] Patent Document 2 reports that a compound called SCL1 has a stomatal opening regulating effect.

[0010] Prior art literature

[0011] Patent Literature

[0012] Patent Document 1: International Publication No. 2014 / 142334

[0013] Patent Document 2: Japanese Patent Application Publication No. 2020-055767

[0014] Non-patent literature

[0015] Non-patent document 1: PNAS, January 7, 2014, vol. 111, no. 1, pp. 533-538

[0016] Non-patent document 2: Front Plant Sci, October 30, 2013, vol. 4, Article 440 Summary of the invention

[0017] Problems to be solved by the invention

[0018] The purpose of the present invention is to provide a plant stomatal opening regulator.

[0019] Methods for solving problems

[0020] The inventors of the present invention have repeatedly conducted in-depth studies on the above-mentioned problems and found that each compound represented by the general formula (1) has a stomatal opening regulating effect. The present invention has been completed as a result of further repeated studies based on this finding. That is, the present invention includes the following aspects.

[0021] Item 1. A plant stomatal opening regulator, comprising at least one selected from the group consisting of a compound represented by general formula (1), a salt thereof, and a solvate thereof,

[0022]

[0023] [In the formula, A represents a ring which may be substituted, L is the same or different and represents a connecting group, and n represents an integer of 1 to 6.]

[0024] Item 2. The plant stomatal opening regulator according to Item 1, wherein the above-mentioned A is a monocyclic ring with 5 to 6 members.

[0025] Item 3. A plant stomatal opening regulator as described in Item 1 or Item 2, wherein the above-mentioned A is an unsubstituted ring, or a ring substituted by at least one selected from a substituted alkyl group, a substituted alkoxy group, a substituted alkoxycarbonyl group, a substituted aryl group, a cyano group, a halogen atom, a nitro group, a substituted amino group, a substituted amide group, a substituted keto group, a substituted ammonium group and a substituted sulfhydryl group.

[0026] Item 4. The plant stomatal opening regulator according to any one of Items 1 to 3, wherein the number of substituents of the above-mentioned A is 0 to 3.

[0027] Item 5. The plant stomatal opening regulator according to any one of Items 1 to 4, wherein L is an alkylene group which may be substituted with an alkyl group and / or an aryl group.

[0028] Item 6. The plant stomatal opening regulator according to any one of Items 1 to 5, wherein

[0029] The above A is a monocyclic ring with 5 to 6 members,

[0030] The above L is an alkylene group which may be substituted by an alkyl group and / or an aryl group,

[0031] When A is a benzene ring and n is 1, A is a benzene ring substituted by at least one selected from a substituted aryl group, a cyano group, an iodine atom and a substituted alkoxycarbonyl group, and / or L is an alkylene group substituted by an alkyl group and / or an aryl group.

[0032] Item 7. The plant stomatal opening regulator according to any one of Items 1 to 6, which is a plant stomatal opening inhibitor.

[0033] Item 8. A drought resistance enhancing agent comprising the plant stomatal opening regulator according to any one of Items 1 to 7.

[0034] Item 9. The drought resistance improving agent according to Item 8, which is used for inhibiting wilting.

[0035] Item 10. A method for improving drought resistance, comprising the step of applying the plant stomatal opening regulator according to any one of Items 1 to 7 to a plant.

[0036] Item 11. The method for improving drought resistance according to Item 10, comprising the step of bringing the plant stomatal opening regulator according to any one of Items 1 to 7 into contact with plant stomata.

[0037] Item 12. A cell membrane proton pump phosphorylation inhibitor comprising at least one selected from the group consisting of a compound represented by general formula (1), a salt thereof, and a solvate thereof.

[0038]

[0039] [In the formula, A represents a ring which may be substituted, L represents a linking group, and n represents an integer of 1 to 6.]

[0040] Item 13. A plant covering or supporting material, comprising at least one selected from the group consisting of a compound represented by the general formula (1), a salt thereof, and a solvate thereof,

[0041]

[0042] [In the formula, A represents a ring which may be substituted, L represents a linking group, and n represents an integer of 1 to 6.]

[0043] Item 14. A compound represented by the general formula (1X), a salt thereof, and a solvate thereof,

[0044]

[0045] [In the formula, A' represents a non-aromatic ring which may be substituted, L is the same or different and represents a connecting group, and n' represents an integer of 2 to 6.]

[0046] Each item specifying the above-mentioned preparation can also be converted into the use of the active ingredient of the present invention in the manufacture of the preparation, the use of the active ingredient of the present invention in the use of the preparation, the active ingredient of the present invention used in the use of the preparation, etc.

[0047] Effects of the Invention

[0048] According to the present invention, a plant stomatal opening regulator and a compound as an active ingredient of the agent can be provided. In addition, a drought resistance enhancer, a cell membrane proton pump phosphorylation inhibitor, and the like can also be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 The measurement results of Test Example 2 are shown. The vertical axis represents the stomatal opening. On the horizontal axis, Dark represents the dark treatment group, Light represents the light treatment group, Ctl represents the case where a test solution containing no test compound is used, ABA represents the case where a test solution containing abscisic acid as a test compound is used, BITC represents the case where a test solution containing BITC (Compound 1) as a test compound is used, and +FC represents the case where a test solution containing fusinosporin in addition to the test compound is used.

[0050] Figure 2 The results of the test example 3 are shown. The vertical axis represents the signal intensity of the phosphorylated cell membrane proton pump. On the horizontal axis, Ctrl represents the case where the test compound is not applied, BITC, m-Bis-BITC and Tris-BITC represent the cases where the test compounds are applied, R represents the case where only red light is applied, RB represents the case where red light and blue light are applied, and FC represents the case where fusosporin is applied.

[0051] Figure 3 The results of the test example 4 are shown. In the upper part of the photo, Ctrl indicates the case where the test compound is not acted on, BITC indicates the case where BITC (Compound 1) is acted on, and ABA indicates the case where abscisic acid is acted on. On the right side of the photo, the detected proteins are shown, 61kDa indicates an ABA-responsive kinase substrate, and 14-3-3 indicates a 14-3-3 protein as a loading control.

[0052] Figure 4The results of the measurement of stomatal aperture after 3 hours of incubation in Test Example 5 are shown. The vertical axis represents stomatal aperture. On the horizontal axis, Ctrl represents the case where a test solution without the test compound is used, BITC and m-Bis-BITC represent the cases where a test solution containing each test compound is used, and the numerical value represents the concentration of the test compound in the test solution (unit: μM).

[0053] Figure 5 The results of stomatal aperture measurement after 3 hours or 48 hours of incubation in Test Example 5 are shown. The vertical axis represents stomatal aperture. On the horizontal axis, the numbers represent the incubation time, and ABA, BITC, and m-Bis-BITC respectively represent the cases where test solutions containing each test compound were used. The leftmost column represents the case where a test solution containing no test compound was used. The concentrations of the test compounds in the test solution are: 100 μM for ABA, 2500 μM for BITC, and 50 μM for m-Bis-BITC.

[0054] Figure 6 The photographic images of the appearance of Test Example 6 are shown. Ctrl indicates the case where a test solution without the test compound is used, BITC and m-Bis-BITC respectively indicate the case where a test solution containing each test compound is used, and the numerical value indicates the concentration of the test compound in the test solution. The upper part shows the photographic images of the appearance at the beginning of the incubation in a dehydrated state, and the lower part shows the photographic images of the appearance at the end of the incubation in a dehydrated state.

[0055] Figure 7 The photographic images of the appearance of Test Example 7 are shown. Ctrl indicates the case where a test solution without the test compound is used, and m-Bis-BITC indicates the case where a test solution containing each test compound is used. The upper half shows the photographic images of the appearance at the beginning of the culture under the water-deficient state, and the lower half shows the photographic images of the appearance at the end of the culture under the water-deficient state.

[0056] Figure 8 The compound 27 synthesized in Test Example 1-1-3 1 HNMR (500 MHz, CDCl3) results. DETAILED DESCRIPTION

[0057] In this specification, the expressions “comprise” and “include” include concepts such as “comprises”, “includes”, “essentially comprises” and “consists of…”.

[0058] The present invention, in one embodiment, relates to a plant stomatal opening regulator (sometimes referred to as "the plant stomatal opening regulator of the present invention" in this specification), which contains at least one selected from the group consisting of a compound represented by general formula (1) (sometimes referred to as "the compound of the present invention" in this specification), a salt thereof, and a solvate thereof (sometimes referred to as "the active ingredient of the present invention" in this specification). The compound of the present invention, the active ingredient of the present invention, and their uses are described below.

[0059]

[0060] 1. Active ingredients

[0061] A represents a ring which may be substituted. It should be noted that the term "substituted" in the present specification does not include substitution with -L-N=C=S.

[0062] The ring is not particularly limited, and includes any of an aromatic ring consisting only of hydrocarbons, an aromatic ring containing heteroatoms other than carbon and hydrogen (heterocycle), a non-aromatic ring consisting only of hydrocarbons (alicyclic hydrocarbons), and a non-aromatic ring containing heteroatoms other than carbon and hydrogen (heterocycle). There are no particular limitations on the heteroatoms contained in the heterocycle, and examples thereof include sulfur atoms, nitrogen atoms, and oxygen atoms. The ring includes any of a monocyclic ring (e.g., a 4- to 8-membered ring) and a condensed ring (e.g., a bicyclic ring or a tricyclic ring). Examples of the aromatic ring include monocyclic rings such as benzene, thiophene, thiazole, furan, pyrrole, imidazole, pyrazole, oxazole, isoxazole, triazole, tetrazole, pyridine, pyrazine, pyrimidine, pyridazine, and triazine; bicyclic rings such as naphthalene, benzothiophene, benzofuran, indole, benzimidazole, indazole, benzoxazole, benzothiazole, isobenzofuran, isoindole, purine, quinoline, isoquinoline, quinoxaline, quinazoline, cinnoline, and phthalazine; and tricyclic rings such as anthracene and phenanthrene. Examples of the non-aromatic ring include cycloalkanes such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, cycloundecane, and cyclododecane; cycloalkenes such as cyclopropene, cyclobutene, cyclopentene, cyclohexene, cycloheptene, and cyclooctene; bicycloalkanes such as bicycloundecane and decalin; and bicycloalkenes such as bicycloundecane and decalin.

[0063] From the viewpoint of regulating the opening of plant stomata, the ring is preferably a monocyclic ring. From the viewpoint of regulating the opening of plant stomata, the ring is preferably a 5- to 6-membered ring. In the case where the ring is a heterocyclic ring, from the viewpoint of regulating the opening of plant stomata, the heteroatom contained in the heterocyclic ring preferably contains a sulfur atom, and more preferably only a sulfur atom. From the viewpoint of regulating the opening of plant stomata, the ring is preferably an aromatic ring.

[0064] The ring is an unsubstituted ring or a ring in which a hydrogen atom is substituted. There are no particular restrictions on the substituent of the ring, and examples thereof include substituted alkyl, substituted alkoxy, substituted alkoxycarbonyl, substituted aryl, cyano, halogen atom, nitro, substituted amino, substituted amide, substituted keto, substituted ammonium, substituted sulfenyl, etc. Among these, substituted alkyl, substituted alkoxy, substituted alkoxycarbonyl, substituted aryl, cyano, halogen atom are more preferred, and substituted aryl, cyano, iodine atom, substituted alkoxycarbonyl are particularly preferred.

[0065] The alkyl group includes any of a straight chain, a branched chain, and a cyclic group. From the viewpoint of regulating the opening of plant stomata, the alkyl group is preferably a straight chain or a branched chain, and more preferably a straight chain. The number of carbon atoms in the alkyl group is not particularly limited, and is, for example, 1 to 8. From the viewpoint of regulating the opening of plant stomata, it is preferably 1 to 6, more preferably 1 to 4, further preferably 1 to 2, and particularly preferably 1. Specific examples of the alkyl group include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, n-hexyl, 3-methylpentyl, and the like. The alkyl group may be substituted by a halogen atom such as a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

[0066] The alkoxy group includes any of a straight chain and a branched chain. From the viewpoint of regulating the opening of plant stomata, the alkoxy group is preferably a straight chain. The carbon number of the alkoxy group is not particularly limited, for example, 1 to 8, and from the viewpoint of regulating the opening of plant stomata, it is preferably 1 to 6, more preferably 1 to 4, further preferably 1 to 2, and particularly preferably 1. Specific examples of the alkoxy group include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, etc. The alkoxy group may be substituted by a halogen atom such as a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, or the like.

[0067] The alkoxycarbonyl group is a group in which an alkoxy group and a carbonyl group are linked together. The alkoxy group is as described above.

[0068] The aryl group is not particularly limited. From the perspective of regulating the opening of plant stomata, it is preferably an aryl group with 6 to 12 carbon atoms, and more preferably an aryl group with 6 to 8 carbon atoms. The aryl group may be any of a monocyclic or polycyclic group (e.g., a bicyclic group, a tricyclic group, etc.). From the perspective of regulating the opening of plant stomata, it is preferably a monocyclic group. As the aryl group, specifically, for example, phenyl, naphthyl, biphenyl, cyclopentadienyl, indenyl, anthracenyl, tetraphenyl, pentaphenyl, pyrenyl, perylenyl, fluorenyl, phenanthrenyl, etc. can be cited, preferably phenyl. The aryl group may be substituted by halogen atoms such as fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms.

[0069] The halogen atom is not particularly limited, and examples thereof include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0070] The hydrogen atoms of the amino group (-NH2), amide group (-CO-NH2), keto group (-CO-H), ammonium group (-NH3) and thiol group (-SH) may be substituted by hydrocarbon groups such as alkyl groups and aryl groups. The alkyl group and aryl group are as described above.

[0071] As the substituent of the ring, an aryl group which may be substituted is particularly preferred from the viewpoint of the effect of regulating plant stomatal opening.

[0072] The number of ring substituents (substituents other than -L-N=C=S) is not particularly limited, and is, for example, 0 to 6. From the viewpoint of the plant stomatal opening regulating effect, the number of substituents is preferably 0 to 3, more preferably 0 to 2, and particularly preferably 0 to 1.

[0073] In the compound of the present invention, in the optionally substituted ring represented by A, the hydrogen atoms on the ring are substituted by n (1 to 6) -L-N=C=S. Thus, the compound of the present invention can exert an excellent plant stomatal opening regulating effect.

[0074] L are the same or different and represent a linking group.

[0075] The connecting group includes a chain structure. As a chain structure, there is no particular limitation, and examples thereof include a chain structure having a main chain constituent atom number of 1 to 20, and from the viewpoint of regulating the opening of plant stomata, examples thereof include preferably 1 to 15, more preferably 1 to 12, further preferably 1 to 10, further preferably 1 to 8, particularly preferably 1 to 6, and particularly preferably 1 to 2. As main chain constituent atoms, for example, carbon atoms, oxygen atoms, sulfur atoms, nitrogen atoms, etc. can be listed. Part or all of the chain structure can be an alkylene group or a heteroalkylene group. The chain structure can be either a straight chain or a branched chain, and from the viewpoint of regulating the opening of plant stomata, a straight chain is preferred. In addition, the chain structure can include local structures such as -O-, -C(=O)-O-, -CO-NH-, -C(=O)-, -NH-, -S(=O)2-, etc. in the main chain. The chain structure can be substituted with an alkyl group and / or an aryl group. The alkyl group and the alkylene group are the same as described above.

[0076] From the viewpoint of the effect of regulating plant stomatal opening, the linking group is preferably an alkylene group which may be substituted with an alkyl group and / or an aryl group, and particularly preferably an alkylene group substituted with an alkyl group and / or an aryl group.

[0077] n represents an integer of 1 to 6.

[0078] From the viewpoint of regulating plant stomatal opening, n is particularly preferably 2 or more. In addition, the upper limit of n is, for example, 5, 4, or 3. In one embodiment of the present invention, from the viewpoint of regulating plant stomatal opening, n is preferably 2 to 5, more preferably 2 to 4, and even more preferably 2 to 3. In addition, from the viewpoint of regulating plant stomatal opening when acting on intact leaves, n is particularly preferably 2.

[0079] In one embodiment of the present invention, it is preferred that A is a monocyclic 5- to 6-membered ring, L is an alkylene group which may be substituted with an alkyl group and / or an aryl group,

[0080] When A is a benzene ring and n is 1, A is a benzene ring substituted by at least one selected from a substituted aryl group, a cyano group, an iodine atom and a substituted alkoxycarbonyl group, and / or L is an alkylene group substituted by an alkyl group and / or an aryl group.

[0081] In one embodiment of the present invention, in the general formula (1), the general formula (1X) is preferably used.

[0082]

[0083] A' represents a non-aromatic ring which may be substituted. L is the same or different and represents a linking group, and n' represents an integer of 2-6.

[0084] In one embodiment of the present invention, in the general formula (1), preferably, the general formula (1Y) is:

[0085]

[0086] In one embodiment of the present invention, in the general formula (1), preferably, the general formula (1A) is:

[0087]

[0088] A" is a benzene ring or cyclohexane. In one embodiment, A" is a benzene ring, and in another embodiment, it is cyclohexane.

[0089] L 1 The same or different groups represent an alkylene group which may be substituted with an alkyl group and / or an aryl group.

[0090] R 1The same or different groups represent an alkyl group which may be substituted, an alkoxy group which may be substituted, an alkoxycarbonyl group which may be substituted, an aryl group which may be substituted, a cyano group, a halogen atom, a nitro group, an amino group which may be substituted, an amide group which may be substituted, a keto group which may be substituted, an ammonium group which may be substituted, or a thiol group which may be substituted. Among them (in one embodiment, when A" is a benzene ring, or when A" is a benzene ring and n is 1), an aryl group which may be substituted, a cyano group, an iodine atom and an alkoxycarbonyl group which may be substituted are particularly preferred, and an aryl group which may be substituted is particularly preferred.

[0091] L 1 and R 1 The groups shown are as described above.

[0092] nSame as above.

[0093] m represents a number satisfying 0≤m≤6-n. From the viewpoint of the plant stomatal opening regulating effect, m is preferably 0-3, more preferably 0-2, and particularly preferably 0-1.

[0094] In one embodiment of the present invention, in the general formula (1), preferably, the general formula (1Z) is:

[0095]

[0096] In one embodiment of the present invention, in the general formula (1), preferably, the general formula (1B) is:

[0097]

[0098] A" is a benzene ring or cyclohexane. In one embodiment, A" is a benzene ring, and in another embodiment, it is cyclohexane.

[0099] R 11 , R 12 , R 13 , R 14 and R 15 may be the same or different, and represent a hydrogen atom, -L-N=C=S, an alkyl group which may be substituted, an alkoxy group which may be substituted, an alkoxycarbonyl group which may be substituted, an aryl group which may be substituted, a cyano group, a halogen atom, a nitro group, an amino group which may be substituted, an amide group which may be substituted, a keto group which may be substituted, an ammonium group which may be substituted, or a thiol group which may be substituted. Among them, an aryl group which may be substituted, a cyano group, an iodine atom, an alkoxycarbonyl group which may be substituted, and an aryl group which may be substituted are particularly preferred.

[0100] L are the same or different and represent a linking group.

[0101] L 1 , R 11 , R 12 , R13 , R 14 and R 15 The groups shown are the same as described above.

[0102] In one embodiment (Embodiment 1) of the present invention, R 12 and R 13 At least one of them is -L-N=C=S, in R 13 In the case of -L-N=C=S, R 11 , R 12 , R 14 and R 15 At least one of is a group other than a hydrogen atom. In Embodiment 1, it is preferred that R 14 and R 15 In Embodiment 1, it is preferred that R 12 and R 13 One of them is -L-N=C=S and the other is a hydrogen atom. 12 In the case of -L-N=C=S, R 11 is a hydrogen atom. In method 1, R 13 In the case of -L-N=C=S, R 11 It is an alkyl group or -L-N=C=S.

[0103] The compound represented by the general formula (1) may contain stereoisomers and optical isomers, but these are not particularly limited.

[0104] The salt of the compound represented by the general formula (1) is not particularly limited as long as it is an agriculturally acceptable salt. As the salt, any of acidic salts and basic salts can be used. Examples of acidic salts include inorganic acid salts such as hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates; organic acid salts such as acetates, propionates, tartrates, fumarates, maleates, malates, citrates, methanesulfonates, and p-toluenesulfonates; examples of basic salts include alkali metal salts such as sodium salts and potassium salts; and alkaline earth metal salts such as calcium salts and magnesium salts; salts with ammonia; salts of morpholine, piperidine, pyrrolidine, monoalkylamines, dialkylamines, trialkylamines, mono(hydroxyalkyl)amines, di(hydroxyalkyl)amines, tri(hydroxyalkyl)amines, etc. with organic amines, etc.

[0105] The compound represented by the general formula (1) may form a hydrate or a solvate. Examples of the solvent include agriculturally acceptable organic solvents (eg, ethanol, glycerol, acetic acid, etc.).

[0106] As the compound represented by the general formula (1), for example, a known compound can be used (for example, a commercially available product). Examples of known compounds include the compounds shown in Tables 1 to 3. In the tables, -NCS represents an isothiocyanate group (-N=C=S).

[0107] [Table 1]

[0108]

[0109] [Table 2]

[0110]

[0111] [Table 3]

[0112]

[0113] The compound represented by the general formula (1) can be produced, for example, by a method including the step of reacting a compound represented by the general formula (a) (compound a) with thiocyanate.

[0114]

[0115] [In the formula, A, L and n are the same as above. X represents a halogen atom.]

[0116] Alternatively, when X is substituted with an amino group, it can also be produced by reacting with a thione-type sulfur-containing compound such as di(1H-imidazol-1-yl)methylthione.

[0117] As compound a, a commercially available product may be used, or a product synthesized according to a known method may be used.

[0118] Examples of the thiocyanate include potassium thiocyanate and sodium thiocyanate, and potassium thiocyanate is preferably used.

[0119] From the viewpoint of yield, ease of synthesis, etc., the amount of thiocyanate used is usually preferably 3 to 30 mol, more preferably 7 to 15 mol, based on 1 mol of compound a.

[0120] The reaction is usually carried out in the presence of a reaction solvent. There are no particular restrictions on the reaction solvent, and examples thereof include dimethylformamide, dichloromethane, acetonitrile, tetrahydrofuran, acetone, toluene, etc., preferably dimethylformamide, etc. The solvent may be used alone or in combination.

[0121] The reaction may be carried out under heating, room temperature or cooling, and is usually carried out at 0 to 120° C. The reaction temperature is particularly preferably 70 to 100° C. The reaction time is not particularly limited, and is usually 30 minutes to 60 hours.

[0122] The progress of the reaction can be tracked by conventional methods such as chromatography. After the reaction is completed, the solvent can be distilled off and separated and purified by conventional methods such as chromatography and recrystallization. In addition, the structure of the product can be determined by elemental analysis, MS (ESI-MS) analysis, IR analysis, 1 H-NMR, 13 Identification by C-NMR, etc.

[0123] 2. Purpose

[0124] The active ingredient of the present invention has a plant stomatal opening regulating effect. Moreover, by regulating the stomatal opening, it is also possible to achieve photosynthesis regulation and thus plant growth regulation. Therefore, at least one selected from the compound represented by the general formula (1), its salt and its solvent can be used as an active ingredient of a photosynthesis regulator, a plant growth regulator and the like.

[0125] In particular, the compound represented by the general formula (1) has the effect of inhibiting the stomatal opening of plants (especially the stomatal opening caused by light or drugs (such as fusin, etc.)). In addition, it is known that by inhibiting the stomatal opening of plants, transpiration can be inhibited and the water content in the plants can be maintained. Therefore, at least one selected from the compound represented by the general formula (1), its salt and its solvent can be used as an active ingredient for plant stomatal opening inhibitors, drought resistance enhancers, wilting inhibitors, preservatives, etc. In addition, it is known that pathogenic microorganisms enter from the opening of plant stomata. Therefore, at least one selected from the compound represented by the general formula (1), its salt and its solvent can be used as an active ingredient for disease resistance enhancers, etc.

[0126] In addition, inhibition of phosphorylation of cell membrane proton pumps is considered to be part of the stomatal opening regulation mechanism of the compound represented by general formula (1). Therefore, at least one selected from the compound represented by general formula (1), its salt and its solvent can be used as an active ingredient of a cell membrane proton pump phosphorylation inhibitor. The cell membrane proton pump used as the object is not particularly limited, as long as it is expressed in guard cells. For example, in the case of Arabidopsis, it is AHA1 (AT2G18960), AHA2 (AT4G30190), AHA3 (AT5G57350), AHA4 (AT3G47950), AHA5 (AT2G24520), AHA6 (AT2G07560), AHA7 (AT3G60330), AHA8 (AT3G42640), AHA9 (AT1G80660), AHA10 (AT1G17260), AHA11 (AT5G62670), etc. In addition, in the case of rice, it is OSA1 (LOC_Os03g48310), OSA2 (LOC_Os07g09340), OSA3 (LOC_Os12g44150), OSA4 (LOC_Os05g25550), OSA5 (LOC_Os08g14360), OSA6 (LOC_Os02g55400), OSA7 (LOC_Os04g56160), OSA8 (LOC_Os03g01120), OSA9 (LOC_Os03g08560), OSA10 (LOC_Os06g08310), etc.

[0127] The target plants of the plant stomatal opening regulator of the present invention are not particularly limited, as long as they are plants with stomata. For example, it can be widely applied to plants such as angiosperms (dicots, monocots, etc.), gymnosperms, ferns, etc. As specific examples, there can be listed: Solanaceae plants such as tomatoes, green peppers, peppers, and eggplants; Cucurbitaceae plants such as cucumbers, pumpkins, melons, and watermelons; Cruciferae leafy vegetables such as cabbage, broccoli, and Chinese cabbage; lettuce or spice vegetables such as celery, coriander, and lettuce, and Allium plants such as green onions, onions, and garlic; Leguminosae plants such as soybeans, peanuts, beans, peas, and red beans; other fruits and vegetables such as strawberries; taproots such as white radish, turnips, carrots, and burdocks; tubers such as taro, cassava, potatoes, sweet potatoes, and yams; asparagus, Soft leafy vegetables such as spinach and celery; flowers such as lisianthus, violets, carnations, and chrysanthemums; cereals such as rice, wheat, barley, oats, and corn; grasses such as bentgrass and sedge; oil crops such as rapeseed and peanuts; sugar crops such as sugarcane and beets; fiber crops such as cotton and rush; fodder crops such as clover, sorghum, and dent corn; deciduous fruit trees such as apples, pears, grapes, and peaches; citrus fruits such as tangerines, lemons, and grapefruits; woody plants such as rhododendron, azalea, and cedar, etc.

[0128] The plant stomatal opening regulator of the present invention may be composed only of the above-mentioned agents, or may contain various additives in addition to the above-mentioned agents according to the dosage form, application method, etc. described later. The content ratio of the above-mentioned agents in the plant stomatal opening regulator can be appropriately determined according to the dosage form, application method, etc. described later, and for example, the range of 0.0001 to 100% by mass can be exemplified. As a more specific example, when the plant stomatal opening regulator of the present invention of the liquid formulation is brought into contact with the stomata, the content ratio of the above-mentioned agents can be exemplified by 1 to 5000 μM, preferably 5 to 3000 μM, more preferably 10 to 1000 μM, further preferably 20 to 500 μM, and further preferably about 20 to 200 μM.

[0129] The formulation of the plant stomatal opening regulator of the present invention is not particularly limited as long as it is an agriculturally acceptable formulation, for example, liquid preparations, solid preparations, powders, granules, fine granules, hydrates, suspensions, emulsions, pastes, dispersants, etc. may be mentioned.

[0130] The additives are not particularly limited as long as they are agriculturally acceptable additives, and examples thereof include carriers, surfactants, spreaders, spray aids, thickeners, fillers, binders, vitamins, antioxidants, pH adjusters, volatilization inhibitors, pigments, and the like.

[0131] Among these, plant water repellency inhibitors such as silicone surfactants are particularly preferred. The content of the silicone surfactant varies depending on the formulation, and in the case of a liquid formulation, for example, it is 0.01 to 0.1% by mass, preferably 0.02 to 0.05% by mass, relative to 100% by mass of the plant stomatal opening regulator of the present invention.

[0132] The application method of the plant stomatal opening regulator of the present invention is not particularly limited as long as it is a known method (or a method to be developed in the future) for the use of pesticides. For example, spraying, dripping, coating, mixing or dissolving in a plant cultivation environment (soil, water, solid culture medium, liquid culture medium, etc.) can be cited. Since the plant stomatal opening regulator of the present invention acts on stomata, it is preferably applied by contacting the plant stomatal opening regulator of the present invention with the stomata of the plant.

[0133] Furthermore, the active ingredient of the present invention may be contained in a plant covering or supporting material for use. From this viewpoint, the present invention, in one embodiment, relates to a plant covering or supporting material containing the active ingredient of the present invention.

[0134] The plant covering material is a material used to cover part or all of the plant, and there is no particular restriction as long as it is such a material. The shape of the plant covering material is not particularly limited, and is preferably in sheet form. The raw material of the plant covering material is not particularly limited, and can be paper, plastic, etc. The plant covering material specifically includes plant packaging sheets, agricultural sheets (agricultural greenhouse sheets), food preservation sheets (e.g., sheets that can be placed in containers (e.g., lunch boxes) for storing food containing plants), etc.

[0135] The plant support material is a material used to support the plant so that it does not fall over, and there is no particular limitation as long as it is such a material. Typically, the plant support material is used by inserting or holding the stem or root of the plant. The material of the plant support material is not particularly limited, and it can be paper, plastic, etc. As the plant support material, for example, porous bodies such as sponges and fiber aggregates can be listed, and more specifically, solid culture media (such as rock wool pots, etc.), mainly absorbent sponges for flowers such as flower arrangements, etc. can be listed.

[0136] The active ingredient of the present invention may constitute the surface layer of the covering or supporting material, or may be contained in the entire covering or supporting material.

[0137] The active ingredient of the present invention in the plant covering or supporting material can exert its effect on the plant by contacting the covering or supporting material with the plant or by volatilizing from the covering or supporting material and contacting the plant.

[0138] Example

[0139] Hereinafter, the present invention will be described in detail based on Examples, but the present invention is not limited to these Examples.

[0140] Test Example 1: Test 1 for determination of stomatal opening regulation effect

[0141] Compounds 1 to 27 (Tables 4 to 10) were prepared and their stomatal opening regulating effects were measured. Compounds 3, 5 and 27 were synthesized as described below. At least compound 27 is a novel compound. Other compounds were synthesized according to or in accordance with previously reported contents or purchased as commercial products.

[0142] <Test Example 1-1: Synthesis of Compounds>

[0143] <Test Example 1-1-1: Synthesis of Compound 3>

[0144] To a dry round-bottom flask with a stirring bar, di(1H-imidazole-1-yl)methylthione (10 mmol, 5 eq.) and DMF (15 mL) were added. A solution of Et3N (4.8 mmol, 2.4 eq.) and 1,4-phenylenediamine (2.0 mmol, 1.0 eq.) in DMF (5 mL) was added to the flask, and the mixture was stirred at 50°C overnight. The reaction mixture was cooled with H2O (20 mL) and extracted with hexane / EtOAc=1:1 (20 mL×3). The organic layer was washed with 1M HCl aqueous solution and water (3 times). The organic layer was then dried over Na2SO4, filtered and concentrated. The crude product was purified by chromatography on silica gel to obtain compound 3 (221 mg, 1.00 mmol, 50%) as a light yellow solid.

[0145] 1 H NMR (500MHz, CDCl3) δ4.73 (s, 4H), 7.35 (s, 4H); 13 C NMR (126MHz, CDCl3) δ48.37, 127.52, 132.83, 134.59; LRMS (GC-MS, EI): m / z=220, C 10 H8N2S2220[M ﹢ ]Calculated value.

[0146] <Test Example 1-1-2: Synthesis of Compound 5>

[0147]

[0148] KSCN (1.36 g, 14 mmol, 10 equiv), NaI (900 mg, 6 mmol, 4.3 equiv) and 1,3,5-tris(bromomethyl)benzene (498 mg, 1.4 mmol, 1.0 equiv) were added to a 100 ml dry round-bottom flask with a stirring bar. Then, dimethylformamide (10 mL) was added to the flask, and the mixture was stirred at 90°C for 24 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with ether (3×20 mL). The organic layer was dried over MgSO4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (hexane / EtOAc=100:0-70:30) and preparative cycle gel permeation chromatography to obtain 1,3,5-tris(isothiocyanatomethyl)benzene (137 mg, 0.47 mmol, 34%). 1 H NMR (500MHz, CDCl3) δ4.79 (s, 6H), 7.26 (s, 3H); 13C NMR (126MHz, CDCl3) δ48.38, 125.29, 133.89, 136.39; LRMS (GC-MS): m / z=291291[M ﹢ ]Calculated value.

[0149] <Test Example 1-1-3: Synthesis of Compound 27>

[0150] To a 100 ml dry round-bottom flask with a stirring bar, di(1H-imidazol-1-yl)methylthione (1.78 g, 10 mmol, 5 eq) and DMF (15 mL) were added. Then, triethylamine (0.55 mL, 9.6 mmol, 4.8 eq) and a DMF (5 mL) solution of trans-1,4-bis(aminomethyl)cyclohexane (280 mg, 2.0 mmol, 1.0 eq) were added to the flask, and the mixture was stirred at 50°C for 16 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with ethyl acetate (3×20 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel to obtain trans-1,4-bis(isothiocyanatomethyl)cyclohexane (315 mg, 1.39 mmol, 70%) as a white solid.

[0151] 1 The results of H NMR (500 MHz, CDCl3) are shown in Figure 8 . Molecular formula: C 10 H 14 N2S2, exact molecular weight: 226.06, molar molecular weight: 226.36.

[0152] <Test Example 1-2: Measurement Test>

[0153] As a test solution, a stomatal aperture measurement solution (5 mM MES-BTP [pH 6.5], 50 mM KCl, 0.1 mM CaCl2, 0.5% DMSO) was prepared in which the test compounds (Compounds 1 to 27) were dissolved at various concentrations.

[0154] Wrap the rice with grass Commelinabenghalensis ) were inserted into a pot with a mixture of vermiculite and peat moss and cultivated for 2 to 4 weeks in a room with sufficient natural light (room temperature 25±3°C). The pots were placed in a dark room for dark treatment overnight. After dark treatment, leaf discs with a diameter of 4 mm were cut from fully expanded leaves using a punch (Biospy Punch, Kai Medical).

[0155] Each test solution was dispensed into 5 wells of a 96-well plate at 50 μl / well, and the leaf disc was immersed in the wells.﹣2 s ﹣1 The cells were incubated for 3 hours under irradiation with blue light. After incubation, stomatal closure was evaluated by image observation (600 times) under a stereofluorescence microscope (Leica M205FA). The short diameter of about 20 stomata (hereinafter referred to as "stomatal opening") was measured for each leaf disk. The 50% inhibition concentration (IC) of stomatal opening was calculated based on the measured values ​​of stomatal opening at each concentration of each test compound. 50 ).

[0156] The results are shown in Tables 4 to 9. In the tables, -NCS represents an isothiocyanate group (-N=C=S), -Me represents a methyl group, and -Ph represents a phenyl group.

[0157] [Table 4]

[0158]

[0159] [Table 5]

[0160]

[0161] [Table 6]

[0162]

[0163] [Table 7]

[0164]

[0165] [Table 8]

[0166]

[0167] [Table 9]

[0168]

[0169] [Table 10]

[0170]

[0171] Test Example 2: Test 2 for determination of stomatal opening regulation effect

[0172] Compound 1 (BITC: benzyl isothiocyanate) (final concentration in the test solution 100 μM) and abscisic acid (final concentration in the test solution 20 μM) were used as test compounds. In addition to the test compound, a test solution to which fusinosporin (final concentration 10 μM) was added was also prepared. In the measurement test, in addition to light (150 μmol m ﹣2 s ﹣1 Red light and 50 μmol m ﹣2 s ﹣1The experiment was conducted in the same manner as in Experimental Example 1, except that the group was irradiated with blue light and incubated for 3 hours (light irradiation group) and the group was incubated in a dark room for 3 hours (dark treatment group) instead of light irradiation. The average value and standard deviation (SD) were calculated from the measured values.

[0173] The results are shown in Figure 1 It can be seen that compound 1 exhibits a stomatal opening inhibitory effect superior to that of abscisic acid, and exhibits an inhibitory effect on stomatal opening caused by fusinosporin.

[0174] Test Example 3: Analysis of the mechanism of stomatal opening regulation 1

[0175] According to the reported technique (Hayashi et al. (2011) Immunohistochemical detection of blue light-induced phosphorylation of the plasma membrane H + -ATPase instomatal guard cells. Plant Cell Physiol. 52: 1238-1248.) The blue light and fusin phosphorylation of the cell membrane proton pump in the guard cells of the Arabidopsis thaliana epidermis were measured by immunohistochemistry. As the test compounds, compound 1 (BITC), compound 4 (m-Bis-BITC) and compound 5 (Tris-BITC) were used. The concentration of compound 1 was set to 100 μM, the concentration of compound 4 was set to 20 μM, the concentration of compound 5 was set to 20 μM, and the concentration of fusin was set to 10 μM.

[0176] The results are shown in Figure 2 It can be seen that compounds 1, 4 and 5 have an inhibitory effect on the phosphorylation of cell membrane proton pumps induced by blue light and the phosphorylation of cell membrane proton pumps induced by fusosporine.

[0177] Test Example 4: Analysis of the mechanism of stomatal opening regulation 2

[0178] According to the reported technology (Takahashi et al. (2007) Protein Phosphorylation and Binding of a 14-3-3 Protein in Vicia Guard Cells in Response to ABA. Plant and Cell Physiology. 48: 1182-1191.), the phosphorylation of ABA-responsive kinase substrate (AKS) induced by abscisic acid (ABA) in the guard cells of the epidermis of Vicia faba was determined by immunoblotting analysis. As test compounds, compound 1 (BITC) and abscisic acid were used. The concentration of compound 1 was set to 100 μM, and the concentration of abscisic acid was set to 20 μM.

[0179] The results are shown in Figure 3 Compound 1 did not induce phosphorylation of ABA-responsive kinase substrates.

[0180] In addition, further analysis revealed that compound 1 had no effect on ABA-related responses (seed germination inhibition, induction of ABA-responsive genes (RAB18 and RD29B)) in Arabidopsis.

[0181] Test Example 5: Test 3 for determination of stomatal opening regulation effect

[0182] Bouquets of chrysanthemum were purchased and grown under 24°C, 16 h of white light (100 μmol m ﹣2 s ﹣1 ) / 8 h dark photoperiod in water bottles for two days. After the dark period, the leaves were immersed in a test solution (0.033% Makupika (spreading agent, Ishihara Biosciences)) supplemented with the test compound (compound 1 (BITC), compound 4 (m-Bis-BITC) or abscisic acid (ABA)) and exposed to white light (100 μmol m ﹣2 s ﹣1 ) for 3 hours, or incubated for 48 hours with a 16-hour light / 16-hour dark photoperiod (light→dark→light). After incubation, the stomatal aperture was measured in the same manner as in Test Example 1.

[0183] The results are shown in Figure 4 and Figure 5 It is known that compounds 1 and 4 also exhibit stomatal opening inhibitory effects on intact leaves. Furthermore, it is known that the stomatal opening inhibitory effect of compound 4 is excellent in durability.

[0184] Test Example 6: Drought resistance test 1

[0185] After the 3-hour incubation in Experiment 5, the bouquets were dehydrated and heated to 10 μmol m﹣2 s ﹣1 Incubate at 21°C under white light and 35-40% relative humidity for 90 minutes.

[0186] Figure 6 The photographs show the appearance at the beginning and end of incubation in a dehydrated state. It can be seen that compounds 1 and 4 can improve drought resistance and inhibit wilting.

[0187] Test Example 7: Drought resistance test 2

[0188] A test solution (0.033% Makupika (spreading agent, Ishihara Bioscience)) containing compound 4 (m-Bis-BITC) was sprayed on 4-5 week old turnips (Brassica rapa) growing in 110 ml of soil. The concentration of compound 4 in the test solution was 50 μM. The plants were then placed in a water-depleted state and cultured in a greenhouse at 25°C and a relative humidity of 40-50% for 24 hours on a sunny day.

[0189] Figure 7 The photographs show the appearance of the plants at the beginning and end of cultivation under water shortage conditions. It can be seen that compound 4 can improve drought resistance and inhibit wilting after a long period of time.

Claims

1. A plant stomatal opening regulator, characterized in that: Containing at least one selected from the group consisting of a compound represented by general formula (1), a salt thereof, and a solvate thereof, In the formula (1), A represents a ring which may be substituted, L is the same or different and represents a linking group, and n represents an integer of 1 to 6.

2. The plant stomatal opening regulator according to claim 1, characterized in that: The A is a monocyclic ring with 5 to 6 members.

3. The plant stomatal opening regulator according to claim 1 or 2, characterized in that: A is an unsubstituted ring, or a ring substituted by at least one selected from a substituted alkyl group, a substituted alkoxy group, a substituted alkoxycarbonyl group, a substituted aryl group, a cyano group, a halogen atom, a nitro group, a substituted amino group, a substituted amide group, a substituted keto group, a substituted ammonium group and a substituted thiol group.

4. The plant stomatal opening regulator according to any one of claims 1 to 3, characterized in that: The number of substituents of A is 0 to 3.

5. The plant stomatal opening regulator according to any one of claims 1 to 4, characterized in that: The L is an alkylene group which may be substituted with an alkyl group and / or an aryl group.

6. The plant stomatal opening regulator according to any one of claims 1 to 5, characterized in that: A is a monocyclic ring with 5 to 6 members, The L is an alkylene group which may be substituted by an alkyl group and / or an aryl group, When A is a benzene ring and n is 1, A is a benzene ring substituted by at least one selected from a substituted aryl group, a cyano group, an iodine atom and a substituted alkoxycarbonyl group, and / or L is an alkylene group substituted by an alkyl group and / or an aryl group.

7. The plant stomatal opening regulator according to any one of claims 1 to 6, characterized in that: It is a plant stomatal opening inhibitor.

8. A drought resistance enhancing agent, characterized in that: A plant stomatal opening regulator comprising the plant stomatal opening regulator according to any one of claims 1 to 7.

9. The drought resistance improving agent according to claim 8, characterized in that: It is used to inhibit wilting.

10. A method for improving drought resistance, characterized in that: The method comprises the step of applying the plant stomatal opening regulator according to any one of claims 1 to 7 to plants.

11. The method for improving drought resistance according to claim 10, wherein: The method comprises the step of bringing the plant stomatal opening regulator according to any one of claims 1 to 7 into contact with the stomata of the plant.

12. A cell membrane proton pump phosphorylation inhibitor, characterized in that: Containing at least one selected from the group consisting of a compound represented by general formula (1), a salt thereof, and a solvate thereof, In the formula (1), A represents a ring which may be substituted, L represents a linking group, and n represents an integer of 1 to 6.

13. A material for covering or supporting plants, characterized in that: Containing at least one selected from the group consisting of a compound represented by general formula (1), a salt thereof, and a solvate thereof, In the formula (1), A represents a ring which may be substituted, L represents a linking group, and n represents an integer of 1 to 6.

14. A compound represented by the general formula (1X), a salt thereof, and a solvate thereof, In the formula (1X), A' represents a non-aromatic ring which may be substituted, L are the same or different and represent a linking group, and n' represents an integer of 2 to 6.

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