An agent for increasing the content of polyphenols in the leaves of theaceae plants and an agent for increasing the content of theanine

Abstract

This invention provides an agent for increasing the content of polyphenols and theanine in the leaves of Theaceae plants. These agents have no adverse effects on the plant's biological tissues and, through appropriate dispersing or irrigation, can safely increase the amount of catechins and other polyphenols and / or theanine contained in tea plants. The agent for increasing the content of polyphenols and / or theanine in the leaves of Theaceae plants comprises at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

Description

Technical Field

[0001] This invention relates to a polyphenol content enhancer and a theanine content enhancer in the leaves of plants in the Theaceae family. Background Technology

[0002] Many plants contain various functional components such as vitamins, carotenoids, and polyphenols. In recent years, due to increased health awareness, there has been greater attention paid to these functional components in crops. In particular, there is a high demand for agricultural products containing large amounts of functional components with antioxidant activity, such as those that remove free radicals (reactive oxygen species), or amino acids and peptides. These reactive oxygen species are considered one of the causes of cell and tissue damage, promoting cancer, lifestyle-related diseases, and aging. The amino acids and peptides mentioned above are also building blocks of proteins in organisms, functioning as various neurotransmitters. Therefore, attempts are underway to significantly increase the production of useful functional components in plants.

[0003] In particular, the leaves of the tea plant, a member of the Theaceae family, contain catechins, a type of polyphenol. These catechins have antioxidant properties, thus helping to prevent aging and a decline in immune function. In addition, they can be expected to reduce body fat and have an effect on bacteria that cause tooth decay.

[0004] Furthermore, among the functional components contained in the leaves of the tea plant (a member of the Theaceae family), there is theanine, a non-protein amino acid. Theanine is the most abundant amino acid in tea and is considered one of its umami components; tea with high theanine content has a higher market value. Therefore, as a cultivation method to increase theanine content, it is known to use mulching material to cover the tea plants, thus blocking sunlight for a certain period (shading cultivation). For tea, if sunlight is blocked due to mulching cultivation, enzyme activity decreases, and the synthesis of catechins from theanine is inhibited. In addition, theanine has been reported to have relaxing effects, improve sleep, and prevent hypertension.

[0005] As a method for increasing the yield of functional components contained in such plants, Patent Document 1 discloses a technique for increasing the amount of polyphenols by using unsaturated fatty acids with a specified structure.

[0006] Furthermore, Patent Document 2 discloses a fertilizer for improving the quality of tea containing stevia extract as an active ingredient, and also describes an increase in theanine content. Patent Document 3 discloses a cultivation method for tea plants that are given betaine before dormancy or after budding, and also describes an increase in theanine content.

[0007] Existing technical documents

[0008] Patent documents

[0009] Patent Document 1: International Publication No. 2020 / 054630

[0010] Patent Document 2: Japanese Patent Application Publication No. 2006-56761

[0011] Patent Document 3: Japanese Patent Application Publication No. 2006-87323 Summary of the Invention

[0012] The problem that the invention aims to solve

[0013] However, no measures were mentioned for increasing the content of catechins in tea leaves.

[0014] In addition, to increase the theanine in tea leaves, stevia extract and betaine are needed. These substances can only be obtained from specific plants and are difficult to obtain.

[0015] This invention was made in view of the above-mentioned problems, and its purpose is to provide an agent that increases the content of polyphenols and theanine in the leaves of Theaceae plants. These are readily available substances that have no adverse effects on the biological tissues of plants. By appropriately distributing or irrigating the plants, the amount of catechins and other polyphenols and / or theanine contained in tea trees can be safely increased.

[0016] Methods for solving problems

[0017] The first aspect of the present invention is a polyphenol content enhancer for the leaves of Theaceae plants, comprising at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0018] The polyphenol content enhancer in the leaves of the Theaceae plant preferably contains oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0019] The compound selected is at least one of the groups consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts, preferably an unsaturated fatty acid with 18 carbon atoms and without an α-keto alcohol structure.

[0020] The preferred oxo-fatty acid is 13-oxo-9,11-octadecadienoic acid or a salt thereof, or 9-oxo-10,12-octadecadienoic acid or a salt thereof.

[0021] The hydroxy fatty acid is preferably 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid.

[0022] Polyphenol content enhancers for the leaves of Theaceae plants are preferably used as sprays or impregnation agents that come into contact with the stems, leaves or roots of Theaceae plants, or as soil irrigation agents.

[0023] In the polyphenol content enhancer of the leaves of Theaceae plants, the polyphenols are preferably catechins.

[0024] The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as a plant activator containing polyphenols, are preferably 0.0001 ppm to 1 ppm, respectively.

[0025] The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as a plant activator containing polyphenols, are preferably 0.001 ppm to 0.1 ppm, respectively.

[0026] The weight ratio of the oxo-fatty acid or its salt to the hydroxy-fatty acid or its salt is preferably 100 to 5 to 100 relative to the oxo-fatty acid or its salt.

[0027] Preferably, the polyphenol content enhancer comprises 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid, wherein the content ratio of 9-oxo-10,12-octadecadienoic acid to 13-oxo-9,11-octadecadienoic acid is 0.1 to 10 by weight.

[0028] Preferably, the polyphenol content enhancer comprises 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid, wherein the content ratio of 9-oxo-10,12-octadecadienoic acid to 13-oxo-9,11-octadecadienoic acid is 0.3 to 2.5 by weight.

[0029] In the polyphenol content enhancer of the leaves of Theaceae plants, the catechins are preferably epigallocatechin gallate.

[0030] Among the polyphenol content enhancers in the leaves of Theaceae plants, the tea plant is the preferred Theaceae plant.

[0031] The second aspect of the present invention is an agent for increasing the content of theanine in the leaves of plants of the Theaceae family, which comprises at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0032] The theanine content enhancer in the leaves of the Theaceae plant preferably contains oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0033] The compound selected is at least one of the groups consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts, preferably an unsaturated fatty acid with 18 carbon atoms and without an α-keto alcohol structure.

[0034] The preferred oxo-fatty acid is 13-oxo-9,11-octadecadienoic acid or a salt thereof, or 9-oxo-10,12-octadecadienoic acid or a salt thereof.

[0035] The hydroxy fatty acid is preferably 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid.

[0036] The theanine content enhancer in the leaves of Theaceae plants is preferably used as a spray or impregnation agent that comes into contact with the stems, leaves or roots of Theaceae plants, or as a soil irrigation agent.

[0037] The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as a plant activator containing theanine as an increaser, are preferably 0.0001 ppm to 1 ppm, respectively.

[0038] The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as a plant activator containing theanine as an increaser, are preferably 0.001 ppm to 0.1 ppm, respectively.

[0039] The weight ratio of the oxo-fatty acid or its salt to the hydroxy-fatty acid or its salt is preferably 100 to 5 to 100 relative to the oxo-fatty acid or its salt.

[0040] Preferably, the theanine content enhancer comprises 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid, wherein the content ratio of 9-oxo-10,12-octadecadienoic acid to 13-oxo-9,11-octadecadienoic acid is 0.1 to 10 by weight.

[0041] Preferably, the ratio of the content of 9-oxo-10,12-octadecadienoic acid to that of 13-oxo-9,11-octadecadienoic acid is 0.3 to 2.5 by weight.

[0042] Among the agents that increase the theanine content in the leaves of plants in the Theaceae family, the tea plant is the preferred plant in the Theaceae family.

[0043] The theanine content enhancer is preferably used in the covered cultivation of Theaceae plants.

[0044] Invention Effects

[0045] The polyphenol content enhancer of the leaves of Theaceae plants of the present invention can increase the polyphenol content in the leaves of Theaceae plants even without the use of stress cultivation or polyphenol-rich varieties, by appropriately distributing or irrigating the plants.

[0046] The theanine content enhancer in the leaves of Theaceae plants of the present invention can increase the theanine content in the leaves of Theaceae plants even without stress cultivation or with varieties that have high theanine content, by appropriately distributing or irrigating the plants. Attached Figure Description

[0047] Figure 1 This is a chart showing a comparison of the catechin content in the leaves of tea plants from Example 1 and Comparative Example 1.

[0048] Figure 2 This is a chart showing a comparison of the theanine content in the leaves of tea plants from Example 2 and Comparative Example 2. Detailed Implementation

[0049] (Regarding the polyphenol content increaser in the leaves of the Theaceae plant of the present invention)

[0050] The plant activator in the leaves of the Theaceae plant of the present invention comprises a polyphenol content enhancer in the leaves of the Theaceae plant, wherein the polyphenol content enhancer in the leaves of the Theaceae plant comprises at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0051] By contacting oxo-fatty acids or their salts, or hydroxy-fatty acids or their salts, with a portion of the stems, leaves, or roots of Theaceae plants, the amount of polyphenols contained in Theaceae plants can be increased. It has been confirmed that the increase in the same components as those increased in normally conducted stress cultivation is observed within the plant; therefore, it is believed that the oxo-fatty acids or their salts of the present invention contain substances and / or precursors that, through absorption by Theaceae plants, perform the same function within the plant as molecules that normally function as signals under environmental stress within Theaceae plants. That is, the oxo-fatty acids or their salts, or hydroxy-fatty acids or their salts of the present invention, can enhance the stress tolerance function inherent in Theaceae plants. As a result, the production of polyphenols within Theaceae plants is promoted and / or their decomposition is inhibited, leading to an increase in polyphenols within Theaceae plants.

[0052] The polyphenol content enhancer in the leaves of the Theaceae plants of the present invention preferably contains both oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0053] In particular, the oxo-fatty acids and hydroxy-fatty acids are preferably 18-carbon unsaturated fatty acids that do not have an α-keto alcohol structure. 18-carbon unsaturated fatty acids are metabolically relevant to organisms, have a high affinity for Camellia sinensis plants, and are also found in plants, thus minimizing environmental impact. It should be noted that an α-keto alcohol structure refers to a structure in which an OH group is bonded to the carbon atom adjacent to the carbonyl group. Fatty acids with an α-keto alcohol structure are easily oxidized and lack stability, therefore they are not preferred as fatty acids used in this invention.

[0054] Oxytofatty acids are so-called rare fatty acids, known to be generated as intermediates in the metabolism of unsaturated fatty acids. 13-oxo-9,11-octadecadienoic acid or 9-oxo-10,12-octadecadienoic acid or their salts, used as examples of oxytofatty acids or salts in this invention, are compounds with a structure in which a carbonyl group with 18 carbon atoms and two double bonds form a conjugated system. 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid are oxytofatty acids generated from linoleic acid, an unsaturated fatty acid, through enzymatic reactions or other means, and are among the rare fatty acids. 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid are known to occur naturally in plants such as tomatoes.

[0055] In addition, hydroxy fatty acids are fatty acids that exist in organisms and are produced by intestinal bacteria such as lactic acid bacteria metabolizing linoleic acid.

[0056] In this invention, 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid are preferably used as hydroxy fatty acids.

[0057] However, it is not known that oxo-fatty acids such as 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid or their salts, or hydroxy fatty acids such as 9,10,13-trihydroxy-11-octadecadienoic acid or 9,12,13-trihydroxy-10-octadecadienoic acid, have the effect of increasing the amount of polyphenols contained in the leaves of Theaceae plants.

[0058] The polyphenol content enhancer for the leaves of Theaceae plants of the present invention can contain any oxo-fatty acid or its salt, or hydroxy fatty acid or its salt, and their sources are not particularly limited. That is, commercially available products can be used as oxo-fatty acids or their salts, or substances contained in plants such as tomatoes can be used directly, or extracted and / or purified. Alternatively, as mentioned above, oxo-fatty acids or their salts, or hydroxy fatty acids or their salts, can be substances obtained by acting an enzyme, such as a plant-derived enzyme, on a matrix such as unsaturated fatty acids, or substances obtained, for example, through chemical synthesis. For example, oxo-fatty acids or their salts, or hydroxy fatty acids or their salts, can be produced by enzymatic conversion of linoleic acid as a raw material through the action of lipoxygenase (LOX) and / or dehydrogenase, such as alcohol dehydrogenase (ADH), or by a catalytic reaction using a metal catalyst. The oxo-fatty acids or their salts obtained in this way can be used to increase the polyphenol content in the leaves of Theaceae plants as needed, at the desired concentration or after appropriate dilution.

[0059] It should be noted that isomers such as (E,E), (Z,E), (E,Z), and (Z,Z) are known to exist in oxo-fatty acids and hydroxy-fatty acids, but these isomers have the same effect as polyphenol content enhancers in the leaves of Theaceae plants. Therefore, in this invention, 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, which can be used as examples of oxo-fatty acids or their salts, or 9,10,13-trihydroxy-11-octadecadienoic acid or 9,12,13-trihydroxy-10-octadecadienoic acid, which can be used as examples of hydroxy-fatty acids or their salts, include all of their isomers. That is, regardless of the isomers of the oxo-fatty acids and hydroxy-fatty acids contained in the polyphenol content enhancer in the leaves of Theaceae plants of this invention, they all exert the same effect as polyphenol content enhancers in the leaves of Theaceae plants.

[0060] In this invention, (9Z,11E)-13-oxo-9,11-octadecadienoic acid and / or (10E,12Z)-9-oxo-10,12-octadecadienoic acid can be used as oxo-fatty acids. Alternatively, 9(S),10(S),13(S)-trihydroxy-11(E)-octadecadienoic acid and / or 9(S),12(S),13(S)-trihydroxy-10(E)-octadecadienoic acid can be used as hydroxy-fatty acids.

[0061] Furthermore, in the polyphenol content enhancer of the leaves of the Theaceae plants of the present invention, it is sufficient to contain oxo-fatty acids or their salts, hydroxy fatty acids or their salts, at the desired concentration. For example, a mixture containing oxo-fatty acids or their salts, hydroxy fatty acids or their salts, can be used as oxo-fatty acids or their salts.

[0062] The polyphenol content enhancer for the leaves of Theaceae plants of the present invention preferably contains water as a solvent. Additionally, pH adjusters, emulsifiers, defoamers, thixotropic agents, antifreeze agents, fertilizer components, etc., may be added to the polyphenol content enhancer as needed.

[0063] The concentrations of oxidized fatty acids or their salts, and hydroxy fatty acids or their salts, contained in the polyphenol content enhancer of the leaves of the Theaceae plants of the present invention, when dispersed as a plant activator containing the polyphenol content enhancer, are preferably 0.0001–1 ppm and 0.0001–1 ppm, respectively, and more preferably 0.001–0.1 ppm and 0.001–0.1 ppm, respectively. This is because, at the above concentrations, oxidized fatty acids or their salts and hydroxy fatty acids or their salts are easily absorbed by the Theaceae plants, and the polyphenol content enhancement effect is at its highest range.

[0064] In cases where both oxo-fatty acids or their salts and hydroxy-fatty acids or their salts are included, the weight ratio of the compound composed of oxo-fatty acids or their salts to the compound composed of hydroxy-fatty acids or their salts is preferably 100 for the compound composed of oxo-fatty acids or their salts and 5 to 100 for the compound composed of hydroxy-fatty acids or their salts. This is because if the content of the compound composed of hydroxy-fatty acids or their salts exceeds 100 by weight relative to 100 for the compound composed of oxo-fatty acids or their salts, the polyphenol-increasing effect may be reduced.

[0065] Furthermore, when 13-oxo-9,11-octadecadienoic acid or its salt and 9-oxo-10,12-octadecadienoic acid or its salt are included as oxo-fatty acids or their salts, the ratio of the content of 9-oxo-10,12-octadecadienoic acid or its salt to the content of 13-oxo-9,11-octadecadienoic acid or its salt, by weight, is in the range of 0.1 to 10, preferably in the range of 0.3 to 2.5, and more preferably in the range of 0.3 to 2.0.

[0066] In the polyphenol content enhancer for the leaves of the Theaceae plants of the present invention, oxidized fatty acids and hydroxy fatty acids may exist in the form of salts. Examples of salts include ammonium salts and metal salts. As metal salts, metal salts that generate monovalent metal ions are preferred, such as sodium salts and potassium salts, but are not limited to these.

[0067] The polyphenol content enhancer for the leaves of Theaceae plants of the present invention is characterized by containing oxidized fatty acids or their salts, or hydroxy fatty acids or their salts, which are natural products. Therefore, it does not cause problems related to soil pollution or toxicity, and can increase the amount of polyphenols contained in Theaceae plants. That is, by using the polyphenol content enhancer of the present invention, the polyphenol content in the leaves of Theaceae plants can be increased safely and easily.

[0068] The polyphenol content enhancer in the leaves of Theaceae plants of the present invention can induce the expression of stress response genes such as PR1, PR2, and PDF1.2 in the applied plants. For example, it can induce the expression of stress response genes such as PR1a and LOXD in Theaceae plants. That is, the polyphenol content enhancer in the leaves of Theaceae plants of the present invention strengthens the stress tolerance function inherent in Theaceae plants. In addition, it can increase the polyphenol content of Theaceae plants even without stress cultivation. It does not cause problems such as reduced yield or reduced resistance to pests and diseases that occur during stress cultivation.

[0069] Furthermore, it is believed that the polyphenol content enhancer in the leaves of Theaceae plants of the present invention can activate genes encoding enzymes required for the biosynthesis of catechins, such as chalcone synthase, chalcone isomerase, and flavonoid synthase. Therefore, it can promote the synthesis of catechins in Theaceae plants and increase the catechin content in the leaves. It should be noted that in the applicant's experiments, the polyphenol content enhancer in the leaves of Theaceae plants of the present invention did not show an effect of increasing caffeine content; therefore, it is believed that the polyphenol content enhancer in the leaves of Theaceae plants of the present invention can selectively activate genes encoding enzymes of the shikimic acid pathway.

[0070] The polyphenol content enhancer for the leaves of Theaceae plants according to the present invention can improve the stress tolerance of Theaceae plants and increase the amount of polyphenols contained in Theaceae plants through simple treatment without changing the conventional cultivation methods.

[0071] The polyphenol content enhancer in the leaves of Theaceae plants of this invention may contain at least one substance selected from amino acids, nucleic acids, and terpenes. This is because these substances have the effect of promoting the growth of Theaceae plants.

[0072] The amino acid can be selected from isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, histidine, tyrosine, cysteine, aspartic acid, asparagine, serine, glutamic acid, glutamine, proline, glycine, alanine, and arginine, or a mixture of two or more of them.

[0073] Additionally, the nucleic acid can be selected from at least one of nucleic acid bases, nucleosides, ribonucleosides, deoxyribonucleosides, ribonucleotides, and deoxyribonucleotides. There is no particular limitation on the nucleic acid; it can be selected from the usual five nucleic acid bases (adenine, guanine, thymine, cytosine, uracil), five ribonucleotides with ribose attached to the nucleic acid bases (adenosine, guanosine, 5-methyluridine, cytidine, uridine), five deoxyribonucleotides with deoxyribose attached to the five nucleic acid bases (deoxyadenosine, deoxyguanosine, deoxythymidine, deoxycytidine, deoxyuridine), or 15 ribonucleotides (AMP (adenosine monophosphate), ADP (adenosine diphosphate), ATP (adenosine triphosphate), GMP (guanosine monophosphate), GDP (guanosine diphosphate), GTP (guanosine triphosphate), TMP (thymidine acid / thymidine)) with 1 to 3 phosphonates attached to the five ribonucleotides. The appropriate choice may be made from 15 deoxyribonucleotides (dAMP, dADP, dATP, dGMP, dGDP, dGTP, dTMP, dTDP, dTTP, dCMP, dCDP, dCTP, dUMP, dUDP, dUTP) in which the 2-hydroxyl group of the ribose of these ribonucleotides is replaced by hydrogen, or modified bases of these nucleic acid bases. Alternatively, a mixture of two or more of these bases may be used.

[0074] As terpenes, any terpene can be suitably used, with monoterpenes, sesquiterpenes, diterpenes, and combinations thereof being preferred. Monoterpenes such as α-pinene, β-pinene, limonene, and others are particularly preferred examples as they are core components of plant hormones. Terpineol may also be included. As isomers of terpineol, α-terpineol, β-terpineol, and γ-terpineol are included, with α-terpineol being more preferred. However, commercially available terpineols are sometimes mixtures of α-terpineol as the main component and a mixture of β-terpineol and γ-terpineol. That is, if α-terpineol is the main component, a mixture of isomers can be used directly. Pine oil, etc., containing α-pinene as the main component, is suitable for use in this invention.

[0075] Examples of polyphenols that increase the amount of components contained in Theaceae plants according to the present invention include anthocyanins, catechins, cocoa polyphenols, rutin, ferulic acid, chlorogenic acid, and curcumin. The tea plant, the most widely cultivated plant in the Theaceae family, contains a large amount of catechins, and the effect of the present invention in increasing polyphenol content is particularly significant among catechins.

[0076] As catechins, various structures are known, including catechin (C), epicatechin (EC), epigallocatechin (EGC), gallocatechin (GC), epicatechin gallate (ECg), catechin gallate (Cg), epigallocatechin gallate (EGCg), and gallocatechin gallate (GCg). In the examples of this specification, the content of epigallocatechin gallate (EGCg) was determined.

[0077] The plants from the Theaceae family to which this invention can be applied are not particularly limited; for example, it can be applied to camellia, camellia flowers, tea (tea tree), summer jasmine, etc.

[0078] Thea plants can be cultivated in any way, either in soil or by immersing them in hydroponic solutions.

[0079] Furthermore, the polyphenol content enhancer for the leaves of Theaceae plants of the present invention can be applied to Theaceae plants by any method. The application method is not particularly limited as long as it involves contact with the roots, stems, leaves, or other parts of the plant. The polyphenol content enhancer for the leaves of Theaceae plants of the present invention can be applied by direct contact with the plant body, or it can be applied to the cultivation carriers such as soil or culture medium used for planting. For example, the polyphenol content enhancer for the leaves of Theaceae plants of the present invention can be used as a spray or impregnation agent for contact with the stems, leaves, or roots of Theaceae plants, or as a soil irrigation agent. Specific application methods can be appropriately selected according to the cultivation morphology of the Theaceae plant to which it is applied. Examples include above-ground liquid dispersal, above-ground solid dispersal, aerial liquid dispersal, aerial solid dispersal, liquid surface dispersal, application within facilities, soil mixing application, soil irrigation application, surface treatment such as coating, application in seedling boxes, single-flower treatment, and stem-root treatment. In addition, the polyphenol content enhancer in the leaves of the Theaceae plants of the present invention can also be mixed with plant fertilizer components and used as a plant fertilizer.

[0080] Furthermore, the polyphenol content enhancer from the leaves of the Theaceae plants of the present invention can also be contained within porous structures or capsules, or impregnated into tablets, etc., and used as a sustained-release agent. The form of the polyphenol content enhancer from the leaves of the Theaceae plants of the present invention is not particularly limited. For example, it can be in liquid or gel form, or in a solid state (block, powder, granules, etc.). In the case of a liquid composition, it can be a concentrated form used directly or diluted.

[0081] The present invention also relates to Theaceae plants cultivated using the above-described cultivation method that exhibit increased polyphenol content. Such Theaceae plants can be utilized using the polyphenols themselves or extracted from Theaceae plants, and are considered useful as food or as raw materials for cosmetics, pharmaceuticals, supplements, etc.

[0082] (Regarding the theanine content increaser in the leaves of the Theaceae plants of the present invention)

[0083] The plant activator of the present invention comprises an agent that increases the content of theanine in the leaves of plants of the Theaceae family, wherein the agent that increases the content of theanine in the leaves of the Theaceae family comprises at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0084] By contacting oxo-fatty acids or their salts, or hydroxy-fatty acids or their salts, with a portion of the stems, leaves, or roots of Theaceae plants, the amount of theanine contained in Theaceae plants can be increased. Since the increase in the same components as those increased in normally conducted stress cultivation can be confirmed within the plant, it is believed that the oxo-fatty acids or their salts of the present invention contain substances and / or precursors that, through absorption by Theaceae plants, perform the same function within the plant as molecules that normally function as signals under environmental stress within Theaceae plants. That is, the oxo-fatty acids or their salts, or hydroxy-fatty acids or their salts of the present invention, can enhance the stress tolerance function inherent in Theaceae plants. As a result, it promotes the production of theanine and / or inhibits its decomposition within Theaceae plants, thereby increasing the theanine content within Theaceae plants.

[0085] The theanine content enhancer in the leaves of the Theaceae plants of the present invention preferably contains both oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

[0086] In particular, the oxo-fatty acids and hydroxy-fatty acids are preferably 18-carbon unsaturated fatty acids that do not have an α-keto alcohol structure. 18-carbon unsaturated fatty acids are metabolically relevant to organisms, have a high affinity for Camellia sinensis plants, and are also found in plants, thus minimizing environmental impact. It should be noted that an α-keto alcohol structure refers to a structure in which an OH group is bonded to the carbon atom adjacent to the carbonyl group. Fatty acids with an α-keto alcohol structure are easily oxidized and lack stability, therefore they are not preferred as fatty acids used in this invention.

[0087] Oxytofatty acids are so-called rare fatty acids, known to be generated as intermediates in the metabolism of unsaturated fatty acids. 13-oxo-9,11-octadecadienoic acid or 9-oxo-10,12-octadecadienoic acid or their salts, used as examples of oxytofatty acids or salts in this invention, are compounds with a structure in which a carbonyl group with 18 carbon atoms and two double bonds form a conjugated system. 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid are oxytofatty acids generated from linoleic acid, an unsaturated fatty acid, through enzymatic reactions or other means, and are among the rare fatty acids. 13-oxo-9,11-octadecadienoic acid and 9-oxo-10,12-octadecadienoic acid are known to occur naturally in plants such as tomatoes.

[0088] In addition, hydroxy fatty acids are fatty acids that exist in organisms and are produced by intestinal bacteria such as lactic acid bacteria metabolizing linoleic acid.

[0089] In this invention, 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid are preferably used as hydroxy fatty acids.

[0090] However, it is not known that oxo-fatty acids such as 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid or their salts, or hydroxy fatty acids such as 9,10,13-trihydroxy-11-octadecadienoic acid or 9,12,13-trihydroxy-10-octadecadienoic acid, have the effect of increasing the amount of theanine contained in the leaves of Theaceae plants.

[0091] The theanine content enhancer for the leaves of Theaceae plants of the present invention can contain any oxo-fatty acid or its salt, or hydroxy fatty acid or its salt, and their sources are not particularly limited. That is, commercially available products can be used as oxo-fatty acids or their salts, or substances contained in plants such as tomatoes can be used directly, or extracted and / or purified. Alternatively, as mentioned above, oxo-fatty acids or their salts, or hydroxy fatty acids or their salts can be substances obtained by acting an enzyme, such as a plant-derived enzyme, on a matrix such as unsaturated fatty acids, or substances obtained, for example, through chemical synthesis. For example, oxo-fatty acids or their salts, or hydroxy fatty acids or their salts can be produced by enzymatic conversion of linoleic acid as a raw material through the action of lipoxygenase (LOX) and / or dehydrogenase, such as alcohol dehydrogenase (ADH), or by a catalytic reaction using a metal catalyst. The oxo-fatty acids or their salts obtained in this way can be used to increase the theanine content in the leaves of Theaceae plants as needed, at the desired concentration or after appropriate dilution.

[0092] It should be noted that oxo-fatty acids and hydroxy-fatty acids are known to exist as isomers such as (E,E), (Z,E), (E,Z), and (Z,Z), and these isomers have the same effect as theanine content enhancers in the leaves of Theaceae plants. Therefore, in this invention, 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, which can be used as examples of oxo-fatty acids or their salts, or 9,10,13-trihydroxy-11-octadecadienoic acid or 9,12,13-trihydroxy-10-octadecadienoic acid, which can be used as examples of hydroxy-fatty acids or their salts, include all of their isomers. That is, regardless of the isomers of the oxo-fatty acids and hydroxy-fatty acids contained in the theanine content enhancer in the leaves of Theaceae plants of this invention, they will all exert the same effect as theanine content enhancers in the leaves of Theaceae plants.

[0093] In this invention, (9Z,11E)-13-oxo-9,11-octadecadienoic acid and / or (10E,12Z)-9-oxo-10,12-octadecadienoic acid can be used as oxo-fatty acids. Alternatively, 9(S),10(S),13(S)-trihydroxy-11(E)-octadecadienoic acid and / or 9(S),12(S),13(S)-trihydroxy-10(E)-octadecadienoic acid can be used as hydroxy-fatty acids.

[0094] Furthermore, in the theanine content enhancer of the leaves of the Theaceae plants of the present invention, it is sufficient to contain oxo-fatty acids or their salts, hydroxy fatty acids or their salts, at the desired concentration. For example, a mixture containing oxo-fatty acids or their salts, hydroxy fatty acids or their salts, can be used as oxo-fatty acids or their salts.

[0095] The theanine content enhancer for the leaves of Theaceae plants of the present invention preferably contains water as a solvent. Additionally, pH adjusters, emulsifiers, defoamers, thixotropic agents, antifreeze agents, fertilizer components, etc., may be added to the theanine content enhancer as needed.

[0096] The concentrations of oxidized fatty acids or their salts, and hydroxy fatty acids or their salts, contained in the theanine content enhancer of the leaves of the Theaceae plants of the present invention, when dispersed as a plant activator containing the theanine content enhancer, are preferably 0.0001–1 ppm and 0.0001–1 ppm, respectively, and more preferably 0.001–0.1 ppm and 0.001–0.1 ppm, respectively. This is because, at the above concentrations, oxidized fatty acids or their salts and hydroxy fatty acids or their salts are easily absorbed by the Theaceae plants, and the theanine content enhancement effect is at its highest range.

[0097] In cases where both oxo-fatty acids or their salts and hydroxy-fatty acids or their salts are included, the weight ratio of the compound composed of oxo-fatty acids or their salts to the compound composed of hydroxy-fatty acids or their salts is preferably 100 for the compound composed of oxo-fatty acids or their salts and 5 to 100 for the compound composed of hydroxy-fatty acids or their salts. This is because if the content of the compound composed of hydroxy-fatty acids or their salts exceeds 100 in weight ratio relative to 100 for the compound composed of oxo-fatty acids or their salts, the effect of increasing theanine may be reduced.

[0098] Furthermore, when 13-oxo-9,11-octadecadienoic acid or its salt and 9-oxo-10,12-octadecadienoic acid or its salt are included as oxo-fatty acids or their salts, the ratio of the content of 9-oxo-10,12-octadecadienoic acid or its salt to the content of 13-oxo-9,11-octadecadienoic acid or its salt, by weight, is in the range of 0.1 to 10, preferably in the range of 0.3 to 2.5, and more preferably in the range of 0.3 to 2.0.

[0099] In the theanine content enhancer of the leaves of the Theaceae plants of the present invention, oxidized fatty acids and hydroxy fatty acids may exist in the form of salts. Examples of salts include ammonium salts and metal salts. As metal salts, metal salts that generate monovalent metal ions are preferred, such as sodium salts and potassium salts, but are not limited to these.

[0100] The theanine content enhancer for the leaves of Theaceae plants of the present invention is characterized by containing oxidized fatty acids or their salts, or hydroxy fatty acids or their salts, which are natural products. Therefore, it does not cause problems related to soil pollution or toxicity, and can increase the amount of theanine contained in Theaceae plants. That is, by using the theanine content enhancer of the present invention, the theanine content in the leaves of Theaceae plants can be increased safely and easily.

[0101] The theanine content enhancer in the leaves of Theaceae plants of the present invention can induce the expression of stress response genes such as PR1, PR2, and PDF1.2 in the applied plants. For example, it can induce the expression of stress response genes such as PR1a and LOXD in Theaceae plants. That is, the theanine content enhancer in the leaves of Theaceae plants of the present invention strengthens the stress tolerance function inherent in Theaceae plants. In addition, it can increase theanine content in Theaceae plants even without stress cultivation. It does not cause problems such as reduced yield or reduced resistance to pests and diseases that occur during stress cultivation.

[0102] Furthermore, it is believed that the theanine content increaser in the leaves of the Theaceae plants of the present invention can activate the genes encoding the enzyme group of the theanine synthesis pathway in Theaceae plants. Therefore, it can promote the synthesis of theanine in Theaceae plants and increase the theanine content in the leaves.

[0103] The theanine content enhancer in the leaves of Theaceae plants according to the present invention can improve the stress tolerance of Theaceae plants and increase the amount of theanine contained in Theaceae plants through simple treatment without changing the conventional cultivation methods.

[0104] The theanine content enhancer in the leaves of Theaceae plants of this invention may contain at least one substance selected from amino acids, nucleic acids, and terpenes. This is because these substances have the effect of promoting the growth of Theaceae plants.

[0105] The amino acid can be selected from isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, histidine, tyrosine, cysteine, aspartic acid, asparagine, serine, glutamic acid, glutamine, proline, glycine, alanine, and arginine, or a mixture of two or more of them.

[0106] Additionally, the nucleic acid can be selected from at least one of nucleic acid bases, nucleosides, ribonucleosides, deoxyribonucleosides, ribonucleotides, and deoxyribonucleotides. There is no particular limitation on the nucleic acid; it can be selected from the usual five nucleic acid bases (adenine, guanine, thymine, cytosine, uracil), five ribonucleotides with ribose attached to the nucleic acid bases (adenosine, guanosine, 5-methyluridine, cytidine, uridine), five deoxyribonucleotides with deoxyribose attached to the five nucleic acid bases (deoxyadenosine, deoxyguanosine, deoxythymidine, deoxycytidine, deoxyuridine), or 15 ribonucleotides (AMP (adenosine monophosphate), ADP (adenosine diphosphate), ATP (adenosine triphosphate), GMP (guanosine monophosphate), GDP (guanosine diphosphate), GTP (guanosine triphosphate), TMP (thymidine acid / thymidine)) with 1 to 3 phosphonates attached to the five ribonucleotides. The appropriate choice may be made from 15 deoxyribonucleotides (dAMP, dADP, dATP, dGMP, dGDP, dGTP, dTMP, dTDP, dTTP, dCMP, dCDP, dCTP, dUMP, dUDP, dUTP) in which the 2-hydroxyl group of the ribose of these ribonucleotides is replaced by hydrogen, or modified bases of these nucleic acid bases. Alternatively, a mixture of two or more of these bases may be used.

[0107] As terpenes, any terpene can be suitably used, with monoterpenes, sesquiterpenes, diterpenes, and combinations thereof being preferred. Monoterpenes such as α-pinene, β-pinene, limonene, and others are particularly preferred examples as they are core components of plant hormones. Terpineol may also be included. As isomers of terpineol, α-terpineol, β-terpineol, and γ-terpineol are included, with α-terpineol being more preferred. However, commercially available terpineols are sometimes mixtures of α-terpineol as the main component and a mixture of β-terpineol and γ-terpineol. That is, if α-terpineol is the main component, a mixture of isomers can be used directly. Pine oil, etc., containing α-pinene as the main component, is suitable for use in this invention.

[0108] The plants from the Theaceae family to which this invention can be applied are not particularly limited; for example, it can be applied to camellia, camellia flowers, tea (tea tree), summer jasmine, etc.

[0109] Thea plants can be cultivated in any way, either in soil or by immersing them in hydroponic solutions.

[0110] Furthermore, the theanine content enhancer in the leaves of the Theaceae plants of the present invention can be applied to Theaceae plants by any method. The application method is not particularly limited as long as it involves contact with the roots, stems, leaves, or other parts of the plant. The theanine content enhancer in the leaves of the Theaceae plants of the present invention can be applied by direct contact with the plant body, or it can be applied to the cultivation carriers such as soil or culture medium in which the plant is planted. For example, the theanine content enhancer in the leaves of the Theaceae plants of the present invention can be used as a spray or impregnation agent for contact with the stems, leaves, or roots of the Theaceae plants, or as a soil irrigation agent. Specific application methods can be appropriately selected according to the cultivation morphology of the Theaceae plants to which the agent is applied. Examples include above-ground liquid dispersal, above-ground solid dispersal, aerial liquid dispersal, aerial solid dispersal, liquid surface dispersal, application within facilities, soil mixing application, soil irrigation application, surface treatment such as coating, application in seedling boxes, single-flower treatment, and stem-root treatment. In addition, the theanine content enhancer in the leaves of the Theaceae plants of the present invention can also be mixed with plant fertilizer components and used as a plant fertilizer.

[0111] Furthermore, the theanine content enhancer in the leaves of the Theaceae plants of the present invention can also be contained in porous structures or capsules, or impregnated in tablets, etc., and used as a sustained-release agent. The form of the theanine content enhancer in the leaves of the Theaceae plants of the present invention is not particularly limited. For example, it can be in liquid or gel form, or in a solid state (block, powder, granules, etc.). In the case of a liquid composition, it can be a concentrated form for direct or diluted use.

[0112] This invention also relates to Theaceae plants with increased theanine content cultivated using the above-described cultivation methods. Such Theaceae plants can be utilized using theanine either directly from the plant or extracted from Theaceae plants, and are considered useful as food or as raw materials for cosmetics, pharmaceuticals, supplements, etc.

[0113] It should be noted that at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts, contained in the polyphenol content enhancers, the theanine content enhancers, and the plant activators disclosed herein contribute to increasing the sugar concentration in tomato leaves. In particular, it contributes to increasing polysaccharides. The activation of photosynthesis is considered one of the main reasons for this increase in sugar concentration.

[0114] Furthermore, at least one compound selected from oxo-fatty acids or their derivatives or salts, and hydroxy fatty acids or their derivatives or salts, contained in the polyphenol content enhancer, the theanine content enhancer, and the plant activator disclosed herein contributes to improving photosynthetic rate and stomatal conductivity. Stomatal conductivity, also known as stomatal conductance, is an indicator of the ease with which gases can pass through stomata. Generally, a high measured value indicates that the stomata are open and photosynthetic activity is active. Photosynthetic rate and stomatal conductivity can be measured using a portable photosynthetic evaporation measurement system.

[0115] Furthermore, in recent years, research on plant factories has flourished, exploring cultivation methods that promote plant growth by increasing light intensity and carbon dioxide concentration within the factory environment. However, under conditions of increased light intensity and carbon dioxide concentration, physiological disorders such as browning and necrosis (so-called tip burn) are sometimes observed in plants, especially leafy vegetables, particularly affecting the leaf margins and sepal tips of new leaves. The polyphenol and theanine content enhancers and plant activators disclosed in this invention contain at least one compound selected from oxo-fatty acids or their derivatives or salts, and hydroxy fatty acids or their derivatives or salts, which helps prevent or inhibit tip burn.

[0116] Example

[0117] The present invention has been described based on embodiments, but the present invention is not limited to the embodiments.

[0118] (Preparation of polyphenol content enhancer)

[0119] As a raw material containing fatty acids, 580g of 90% pure linoleic acid (manufactured by Nippon Oil Co., Ltd.) was used, along with 216g of potassium carbonate (manufactured by Fujifilm Wako Co., Ltd.), 280g of dipotassium hydrogen phosphate (manufactured by Fujifilm Wako Co., Ltd.), and 13000mL of distilled water to prepare the test solution. The pH of the test solution at this time was 9.0.

[0120] Add 40 mg of lipoxygenase (manufactured by Nacalai Tesque, derived from soybean) to the test solution, and react at 15°C for 3 hours while aerating and stirring. Then, place the reaction mixture in a 90°C hot water bath for 90 minutes. The resulting reaction solution is referred to as solution A.

[0121] Add 35 mL of phosphoric acid (manufactured by Fujifilm Wōkō) to 6500 mL of solution A to adjust the pH to 7.0. While aerating the solution and stirring, react at 50°C for 22 hours, then place the reaction mixture in a 90°C hot water bath for 2 hours. The resulting reaction solution is referred to as solution B.

[0122] The total amount of solution B obtained above was mixed with the remaining total amount of solution A that was not used to prepare solution B. For the resulting mixture, 13-oxoODA (13-oxo-9,11-octadecadienoic acid, 13-oxo-9,11-octadecadienoic acid) and 9-oxoODA (9-oxo-10,12-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid) manufactured by Kyman Chemical and 9,10,13-trihydroxy-11-octadecenoic acid and 9,12,13-trihydroxy-10-octadecenoic acid manufactured by Laroda Fine Chemicals were used as standards. The mixture was analyzed using MS2 spectra and quantified using liquid chromatography-mass spectrometry (LC-MS).

[0123] Furthermore, quantification was performed using an absolute calibration curve method for ketoctadecadienoic acid (13-oxoODA, 9-oxoODA) at a detection wavelength of UV 272 nm, and for trihydroxyoctadecadienoic acid at a detection wavelength of UV 210 nm. The total yield of 13-oxoODA, including the (E,E) and (E,Z) isomers, was 3.7%. The yield of 9-oxoODA was 1.7%. Additionally, the combined yield of 9,10,13-trihydroxy-11-octadecadienoic acid and 9,12,13-trihydroxy-10-octadecadienoic acid was 1.2% (the peak could not be separated by LC-MS), and the recovery of linoleic acid was 84.1%.

[0124] The mixture of solutions A and B obtained above (plant activator) was diluted to 2000 mL with deionized water to obtain the plant activation solution. The concentration of 13-oxoODA in the plant activation solution was 0.00875 ppm, the concentration of 9-oxoODA was 0.0040 ppm, and the combined concentration of 9,10,13-trihydroxy-11-octadecenoic acid and 9,12,13-trihydroxy-10-octadecenoic acid was 0.0029 ppm.

[0125] (Example 1)

[0126] (Plant cultivation experiment)

[0127] During the budding stage of the third bud of conventionally cultivated tea trees in the open field (tea variety: Yabukita), 75L of plant activating solution was dispersed over a 250m² area using a boom sprayer. 2 The area.

[0128] (Analysis Methods)

[0129] (Table showing gallocatechin gallate and caffeine levels)

[0130] Approximately three months after the distribution date, ten plants were randomly selected, and new leaves were collected to determine the levels of epigallocatechin gallate (EGCg) and caffeine. The levels of epigallocatechin gallate and caffeine were calculated as follows: Samples were frozen at -80°C for one day, then immersed in a 10-fold mixture of water, acetic acid, and methanol (20:1:80). After 30 minutes of ultrasonic treatment, the samples were allowed to stand at 25°C for 24 hours. The resulting extract was then analyzed. The average values ​​of the epigallocatechin gallate and caffeine content from the ten plants were calculated.

[0131] Comparative Example 1 (Water Treatment)

[0132] In the same tea garden as in Example 1, water was spread over the same area instead of plant activator, and the amount of epigallocatechin gallate (EGCg) and caffeine contained in the new leaves was determined using the same method as in Example 1.

[0133] Analysis was performed using an LC-MS / MS apparatus (LC section: DIONX Ultimate 3000, MS / MS section: Q Exactive Focus: Thermo Fisher Scientific, Inc.) under the following conditions: Column = Aclaim PR-MS 2.1mm Φ × 150mm (Thermo Fisher Scientific, Inc.), solvent = 15% methanol / acetic acid water, flow rate = 0.25 mL / min, column temperature = 40℃, detection UV 280nm, sample solution 2 μL. Caffeine standard (Fujifilm and Koko Pure Chemical Industries, Ltd.) and (-)-epigallocatechin gallate standard (Fujifilm and Koko Pure Chemical Industries, Ltd.) were used as reference materials for quantification.

[0134] (result)

[0135] Depend on Figure 1 It is understandable that by administering polyphenol-enhancing agents to the leaves of Theaceae plants containing 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, 9,10,13-trihydroxy-11-octadecadienoic acid, and 9,12,13-trihydroxy-10-octadecadienoic acid, the catechin content in tea leaves increases. Furthermore, no increase in caffeine content was confirmed in tea leaves; therefore, it can be inferred that this selectively activates genes encoding enzymes in the catechin synthesis pathway.

[0136] (Preparation of theanine content enhancer)

[0137] As a raw material containing fatty acids, 580g of 90% pure linoleic acid (manufactured by Nippon Oil Co., Ltd.) was used, along with 216g of potassium carbonate (manufactured by Fujifilm Wako Co., Ltd.), 280g of dipotassium hydrogen phosphate (manufactured by Fujifilm Wako Co., Ltd.), and 13000mL of distilled water to prepare the test solution. The pH of the test solution at this time was 9.0.

[0138] Add 40 mg of lipoxygenase (manufactured by Nacalai Tesque, derived from soybean) to the test solution, and react at 15°C for 3 hours while aerating and stirring. Then, place the reaction mixture in a 90°C hot water bath for 90 minutes. The resulting reaction solution is referred to as solution A.

[0139] Add 35 mL of phosphoric acid (manufactured by Fujifilm Wōkō) to 6500 mL of solution A to adjust the pH to 7.0. While aerating the solution and stirring, react at 50°C for 22 hours, then place the reaction mixture in a 90°C hot water bath for 2 hours. The resulting reaction solution is referred to as solution B.

[0140] The total amount of solution B obtained above was mixed with the remaining total amount of solution A that was not used to prepare solution B. For the resulting mixture, 13-oxoODA (13-oxo-9,11-octadecadienoic acid, 13-oxo-9,11-octadecadienoic acid) and 9-oxoODA (9-oxo-10,12-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid) manufactured by Kyman Chemical and 9,10,13-trihydroxy-11-octadecenoic acid and 9,12,13-trihydroxy-10-octadecenoic acid manufactured by Laroda Fine Chemicals were used as standards. The mixture was analyzed using MS2 spectra and quantified using liquid chromatography-mass spectrometry (LC-MS).

[0141] Furthermore, quantification was performed using an absolute calibration curve method for ketoctadecadienoic acid (13-oxoODA, 9-oxoODA) at a detection wavelength of UV 272 nm, and for trihydroxyoctadecadienoic acid at a detection wavelength of UV 210 nm. The total yield of 13-oxoODA, including the (E,E) and (E,Z) isomers, was 3.7%. The yield of 9-oxoODA was 1.7%. Additionally, the combined yield of 9,10,13-trihydroxy-11-octadecadienoic acid and 9,12,13-trihydroxy-10-octadecadienoic acid was 1.2% (the peak could not be separated by LC-MS), and the recovery of linoleic acid was 84.1%.

[0142] The mixture of solutions A and B obtained above (plant activator) was diluted to 2000 mL with deionized water to obtain the plant activation solution. The concentration of 13-oxoODA in the plant activation solution was 0.0642 ppm, the concentration of 9-oxoODA was 0.0295 ppm, and the combined concentration of 9,10,13-trihydroxy-11-octadecenoic acid and 9,12,13-trihydroxy-10-octadecenoic acid was 0.0208 ppm.

[0143] (Example 2)

[0144] (Plant cultivation experiment)

[0145] Nineteen days before the harvest of the first tea of ​​conventionally cultivated tea trees in the open field (tea variety of the tea garden: Yabukita), 50L of plant activator solution was spread to area 4a using a boom sprayer, and the area was covered with coarse cotton cloth (shading) until harvest.

[0146] (Analysis Methods)

[0147] (Theanine content)

[0148] At harvest time, 10 tea plants were randomly selected, and new leaves were collected to determine the amount of theanine per unit weight. The theanine content was calculated as follows: the sample was frozen at -80℃ for 1 day, then immersed in a 10-fold mixture of water:acetic acid:methanol = 20:1:80, subjected to ultrasound for 30 minutes, and then allowed to stand at 25℃ for 24 hours. The resulting extract was analyzed to determine the theanine content. The average theanine content of the 10 plants was calculated.

[0149] Comparative Example 2 (Water Treatment)

[0150] In the same tea garden as in Example 1, water was spread over the same area instead of plant activator, and the area was covered with coarse cotton cloth (for shade) until harvest. The amount of theanine in the new leaves was determined using the same method as in Example 2.

[0151] Analysis was performed using an LC-MS / MS apparatus (LC section: DIONX Ultimate 3000, MS / MS section: Q Exactive Focus: Thermo Fisher Scientific, Inc.) under the following conditions: Column = Aclaim PR-MS 2.1mm Φ × 150mm (Thermo Fisher Scientific, Inc.), Solvent = 0% acetonitrile / acetic acid aqueous solution → 30% acetonitrile / acetic acid aqueous solution, Flow rate = 0.25 mL / min, Column temperature = 40°C, Detection = MS-(SIM), Immunosorbent = 2 μL sample solution. L-theanine (Fujifilm and Koichi Pure Chemicals Co., Ltd.) was used as a standard for qualitative analysis.

[0152] Depend on Figure 2 It is understandable that the theanine content in tea leaves increases by administering theanine-enhancing agents to the leaves of Theaceae plants containing 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, 9,10,13-trihydroxy-11-octadecadienoic acid, and 9,12,13-trihydroxy-10-octadecadienoic acid. The activation of theanine synthesis can be inferred by inhibiting the synthesis of catechins from theanine through mulching cultivation (shade cultivation).

[0153] As described above in Example 1, by imparting a plant activator containing 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, 9,10,13-trihydroxy-11-octadecadienoic acid and 9,12,13-trihydroxy-10-octadecadienoic acid, the content of catechins in the leaves of tea plants is increased.

[0154] In addition, as described in Example 2, the content of theanine in tea leaves is increased by imparting a plant activator containing 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, 9,10,13-trihydroxy-11-octadecadienoic acid and 9,12,13-trihydroxy-10-octadecadienoic acid.

[0155] Therefore, plant activators containing 13-oxo-9,11-octadecadienoic acid, 9-oxo-10,12-octadecadienoic acid, 9,10,13-trihydroxy-11-octadecadienoic acid, and 9,12,13-trihydroxy-10-octadecadienoic acid can be used as agents to increase the content of polyphenols and theanine in the leaves of Theaceae plants.

[0156] This application claims priority based on Japanese Patent Application No. 2024-005002 filed on January 17, 2024 and Japanese Patent Application No. 2024-035510 filed on March 8, 2024, the contents of which are incorporated herein by reference.

[0157] The contents of International Publication No. 2020 / 054630, Japanese Patent Application Publication No. 2006-56761 and Japanese Patent Application Publication No. 2006-87323 are incorporated herein by reference.

Claims

1. A polyphenol content enhancer for the leaves of a plant in the Theaceae family, comprising at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

2. The polyphenol content enhancer in the leaves of the Theaceae plant according to claim 1, comprising oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

3. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 1 or 2, wherein, The compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts is an unsaturated fatty acid with 18 carbon atoms and without an α-keto alcohol structure.

4. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 3, wherein, The oxo-fatty acid is 13-oxo-9,11-octadecadienoic acid or 9-oxo-10,12-octadecadienoic acid.

5. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 3, wherein, The hydroxy fatty acid is 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid.

6. The polyphenol content enhancer in the leaves of Theaceae plants according to claim 1 or 2, which is used as a spray or impregnation agent in contact with the stems, leaves or roots of Theaceae plants, or as a soil irrigation agent.

7. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 1 or 2, wherein, The polyphenols are catechins.

8. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 1 or 2, wherein, The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as plant activators containing polyphenols, are respectively between 0.0001 ppm and 1 ppm.

9. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 1 or 2, wherein, The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as plant activators containing polyphenols, are respectively between 0.001 ppm and 0.1 ppm.

10. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 2, wherein, The weight ratio of the oxo-fatty acid or its salt to the hydroxy-fatty acid or its salt is 5 to 100 relative to the oxo-fatty acid or its salt.

11. The polyphenol content enhancer for leaves of Theaceae plants according to claim 4, comprising the 13-oxo-9,11-octadecadienoic acid and the 9-oxo-10,12-octadecadienoic acid, wherein the content ratio of the 9-oxo-10,12-octadecadienoic acid to the 13-oxo-9,11-octadecadienoic acid is 0.1 to 10 by weight.

12. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 11, wherein, The content ratio of 9-oxo-10,12-octadecadienoic acid to 13-oxo-9,11-octadecadienoic acid is 0.3 to 2.5 by weight.

13. The polyphenol content enhancer for the leaves of Theaceae plants according to claim 7, wherein, The catechins mentioned are epigallocatechin gallate esters.

14. The polyphenol content enhancer for the leaves of Theaceae plants according to claims 1-13, wherein, The plant in question is the tea tree (Camellia sinensis).

15. An agent for increasing the theanine content in the leaves of a plant in the Theaceae family, comprising at least one compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

16. The theanine content enhancer in the leaves of the Theaceae plant according to claim 15, comprising oxidized fatty acids or their salts and hydroxy fatty acids or their salts.

17. The theanine content enhancer in the leaves of Theaceae plants according to claim 15 or 16, wherein, The compound selected from the group consisting of oxidized fatty acids or their salts and hydroxy fatty acids or their salts is an unsaturated fatty acid with 18 carbon atoms and without an α-keto alcohol structure.

18. The theanine content enhancer in the leaves of Theaceae plants according to claim 17, wherein, The oxo-fatty acid is 13-oxo-9,11-octadecadienoic acid or 9-oxo-10,12-octadecadienoic acid.

19. The theanine content enhancer in the leaves of Theaceae plants according to claim 17, wherein, The hydroxy fatty acid is 9,10,13-trihydroxy-11-octadecenoic acid or 9,12,13-trihydroxy-10-octadecenoic acid.

20. The theanine content enhancer in the leaves of Theaceae plants according to claim 15 or 16, which is used as a spray or impregnation agent in contact with the stems, leaves or roots of Theaceae plants, or as a soil irrigation agent.

21. The theanine content enhancer in the leaves of Theaceae plants according to claim 15 or 16, wherein, The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as plant activators containing theanine as an increaser, are respectively between 0.0001 ppm and 1 ppm.

22. The theanine content enhancer in the leaves of Theaceae plants according to claim 21, wherein, The concentrations of the oxidized fatty acids or their salts and the concentrations of the hydroxy fatty acids or their salts, when dispersed as plant activators containing theanine as content enhancers, are respectively 0.001 ppm to 0.1 ppm.

23. The theanine content enhancer in the leaves of Theaceae plants according to claim 16, wherein, The weight ratio of the oxo-fatty acid or its salt to the hydroxy-fatty acid or its salt is 5 to 100 relative to the oxo-fatty acid or its salt.

24. The theanine content enhancer in the leaves of Theaceae plants according to claim 18, comprising the 13-oxo-9,11-octadecadienoic acid and the 9-oxo-10,12-octadecadienoic acid, wherein the content ratio of the 9-oxo-10,12-octadecadienoic acid to the 13-oxo-9,11-octadecadienoic acid is 0.1 to 10 by weight.

25. The theanine content enhancer in the leaves of Theaceae plants according to claim 24, wherein, The content ratio of 9-oxo-10,12-octadecadienoic acid to 13-oxo-9,11-octadecadienoic acid is 0.3 to 2.5 by weight.

26. The theanine content enhancer in the leaves of Theaceae plants according to claims 15-25, wherein, The plant in question is the tea tree (Camellia sinensis).

27. The theanine content enhancer according to claims 15-26, wherein, The theanine content enhancer is used in the cover cultivation of Theaceae plants.

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