JP7597848B2 - Plant functional ingredient enhancer - Google Patents

Description

The present invention relates to an agent for increasing functional components of plants.

Many plants contain various functional ingredients such as vitamins, carotenoids, and polyphenols. In recent years, interest in these functional ingredients contained in agricultural crops has increased due to health-conscious trends, and there is a particular need for agricultural products that contain functional ingredients with antioxidant activity that remove radicals such as active oxygen, which are thought to damage cells and tissues and contribute to cancer, lifestyle-related diseases, and accelerated aging, as well as amino acids and peptides that are building blocks of proteins in living organisms and function as various neurotransmitters. Therefore, attempts have been made to significantly increase the production of useful functional ingredients in plants.

As a method for increasing the yield of functional components contained in plants, Patent Document 1 discloses a technique for increasing the amount of polyphenols contained in strawberries by irradiating them with ultraviolet light.

International Publication No. 2012/133130

However, irradiating plants with ultraviolet light can destroy the plant's biological tissue, so there was a limit to the amount of time that it could be exposed to the light. There were also concerns that ultraviolet light could be harmful to the human body, and there was a problem in that it was not possible to fully ensure the safety of workers.

The present invention was made in consideration of the above problems, and aims to provide an agent for increasing the amount of functional components in plants that does not adversely affect the living tissue of plants and can be safely sprayed or irrigated onto plants to increase the amount of functional components contained in the plants.

The present invention relates to a compound of the formula:
HOOC-(R ¹ )-C=CC(=O)-R ² (I)
(Wherein,
R ¹ : a linear or branched alkylene group having 6 to 12 carbon atoms, which may contain one or more double bonds;
^R2 : an alkyl group having 2 to 8 carbon atoms, which may contain one or more branches and/or double bonds.
The present invention relates to an agent for increasing functional components of plants, which comprises, as an active ingredient, an oxo fatty acid derivative having the structural formula:

The agent for increasing functional components for plants is preferably an oxo fatty acid derivative in which the alkylene group of R ¹ has 8 to 10 carbon atoms and the alkyl group of R ² has 4 to 6 carbon atoms.

The agent for increasing functional components of plants is preferably an oxo fatty acid derivative in which R ¹ of the oxo fatty acid derivative contains a double bond that forms a conjugated double bond with the double bond between the α and β carbons of the carbonyl group in formula (I).

The agent for increasing functional components in plants is preferably an oxo fatty acid derivative in which R ¹ is an alkylene group having 9 carbon atoms and R ² is an alkyl group having 5 carbon atoms.

The oxo fatty acid derivative is preferably 13-oxo-9,11-octadecadienoic acid or a salt thereof.

The functional component enhancer for plants is preferably used as a spray or immersion agent that is brought into contact with the stems, leaves, or roots of plants, or as a soil irrigation agent.

Preferably, the functional component of the plant is an antioxidant functional component, amino acids, or peptides.

The functional component of the plant is preferably an agent for increasing the functional components of the plant selected from the group consisting of vitamins, polyphenols, carotenoids, amino acids, and combinations thereof.

The functional component of the plant is preferably at least one selected from the group consisting of vitamin C, polyphenols, lutein, β-carotene, lycopene, and GABA.

The functional component increaser for plants is preferably used for plants of the Solanaceae, Rosaceae, Apiaceae, Lamiaceae, or Amaranthaceae family.

In addition, the functional component enhancer of plants referred to in the present invention is an agent that promotes the production and/or inhibits the decomposition of functional components within plants, thereby increasing the functional components in the plants.

The plant functional component increaser of the present invention can increase the functional components of plants by appropriately spraying or irrigating the plants, without the need for stress cultivation or using high-content varieties.

The functional component increaser for plants of the present invention is
An oxo fatty acid derivative having the following formula:
HOOC-(R ¹ )-C=CC(=O)-R ² (I)
(Wherein,
R ¹ : a linear or branched alkylene group having 6 to 12 carbon atoms, which may contain one or more double bonds;
^R2 : an alkyl group having 2 to 8 carbon atoms, which may contain one or more branches and/or double bonds.
The present invention also relates to a plant functional ingredient enhancer comprising, as an active ingredient, a compound of formula (I) or a salt thereof, including all geometric isomers and stereoisomers.

By contacting the oxo fatty acid derivative or its salt with a part of the stem, leaf, or root of a plant, the amount of functional components contained in the plant can be increased. Since an increase in the same components in the plant that increase in commonly performed stress cultivation can be confirmed, it is believed that the oxo fatty acid derivative or its salt of the present invention contains a substance and/or its precursor that, when absorbed by the plant, acts in the plant in the same way as a molecule that is naturally produced and acts as a signal in the plant in response to environmental stress. In other words, the oxo fatty acid derivative or its salt of the present invention can enhance the stress resistance function that the plant naturally possesses. As a result, the production of functional components in the plant is promoted and/or the decomposition is inhibited, and the functional components in the plant are increased.

Oxo fatty acids are so-called rare fatty acids that are known to be produced as intermediates in the metabolism of unsaturated fatty acids. These rare fatty acids are attracting attention, particularly from the viewpoint of their application in various industrial uses, such as their physiological activity. 13-oxo-9,11-octadecadienoic acid, which is used as an example of an oxo fatty acid derivative or its salt in the present invention, is a compound having 18 carbon atoms and a carbonyl group and two conjugated double bonds in the molecule. It is an oxo fatty acid produced from linoleic acid, an unsaturated fatty acid, by an enzyme reaction or other means, and is one of the rare fatty acids. 13-oxo-9,11-octadecadienoic acid is known to exist naturally in plants such as tomatoes. 13-oxo-9,11-octadecadienoic acid has been found to have activity in improving lifestyle-related diseases, such as improving lipid metabolism, and is therefore being actively researched both domestically and internationally as a functional ingredient that exhibits a remarkable fat-burning effect.

However, it was not known that oxo fatty acid derivatives such as 13-oxo-9,11-octadecadienoic acid or their salts have the effect of increasing the amount of functional components contained in plants.

The plant functional component increaser of the present invention may contain an oxo fatty acid derivative or a salt thereof, and the origin thereof is not particularly limited. That is, the oxo fatty acid derivative or a salt thereof may be a commercially available product, or may be one contained in a plant such as a tomato, and may be used as is, or may be extracted and/or purified. Alternatively, the oxo fatty acid derivative or a salt thereof may be obtained by acting an enzyme, for example an enzyme derived from a microorganism, on a substrate such as an unsaturated fatty acid, as described above, or may be obtained by chemical synthesis, or may be produced using a microorganism. For example, the oxo fatty acid derivative or a salt thereof may be produced by enzymatic conversion using linoleic acid as a raw material by the action of lipoxygenase (LOX) and/or dehydrogenase, for example alcohol dehydrogenase (ADH), or through a catalytic reaction using a metal catalyst. The oxo fatty acid derivative or a salt thereof thus obtained may be used at a desired concentration or appropriately diluted as necessary to increase the functional components in the plant.

As mentioned above, it is known that oxo fatty acids exist in isomers such as (E,E), (Z,E), (E,Z), and (Z,Z), but the effects of these isomers in the plant functional component increaser are similar. Therefore, in the present invention, 13-oxo-9,11-octadecadienoic acid, which can be used as an example of an oxo fatty acid derivative or its salt, is considered to include all of its isomers. In other words, the oxo fatty acid contained in the plant functional component increaser of the present invention exerts the same effect as the plant functional component increaser, regardless of the isomer present in the plant functional component increaser.

The plant functional component increaser of the present invention may contain a desired concentration of an oxo fatty acid derivative or a salt thereof. For example, a mixture containing an oxo fatty acid derivative may be used as the oxo fatty acid derivative or a salt thereof.

In the plant functional component increaser of the present invention, the oxo fatty acid derivative may be present in the form of a salt, and examples of the salt include ammonium salts and metal salts. Metal salts that generate monovalent metal ions are preferred, and for example, sodium salts and potassium salts can be preferably used, although this is not limited thereto.

The plant functional component increaser of the present invention is characterized by containing a natural product, an oxo fatty acid derivative or its salt, and therefore can increase the amount of functional components contained in plants without causing problems related to soil contamination or toxicity. In other words, by using the plant functional component increaser of the present invention, it is possible to safely and easily increase the amount of functional components in plants.

The functional component increaser for plants of the present invention can induce the expression of stress response genes such as PR1, PR2, and PDF1.2 in plants to which it is applied. For example, in the case of a plant of the Solanaceae family, it can induce the expression of stress response genes such as PR1a and LOXD. As a result, depending on the type and variety of the plant, the growth stage, the cultivation environment, and the season, the development of the cuticle, the development of trichomes, the promotion of root hair development, the increase in the amount of antioxidants produced, the promotion of the function to prevent water evaporation (increasing the production of proline and the like and thickening the leaves), the thickening of the stem, and the like occur. That is, the functional component increaser for plants of the present invention strengthens the stress resistance function that plants inherently possess. Therefore, the functional components of plants can be increased without using stress cultivation. Problems such as a decrease in yield and a decrease in resistance to pests that occur when stress cultivation or the use of a high-content variety do not occur. According to the functional component increaser for plants of the present invention, the stress resistance function of plants can be improved by simple processing without changing the conventional cultivation method, and the amount of functional components contained in the plants can be increased.

Examples of functional components whose content is increased by the present invention include vitamins, polyphenols, carotenoids, peptides, and amino acids. Examples of vitamins include vitamin C, and examples of polyphenols include chlorogenic acid, scopoletin, sinapic acid, and sinapaldehyde. Examples of carotenoids include β-carotene, lutein, and lycopene. Examples of peptides and amino acids include GABA, glutamic acid, and peptides formed by dehydration and condensation of these. At least one of these functional components can be increased by the plant functional component increaser of the present invention.

Plants to which the present invention can be applied include, but are not limited to, plants of the Solanaceae, Brassicaceae, Asteraceae, Fabaceae, Liliaceae, Rosaceae, Apiaceae, Lamiaceae, and Amaranthaceae families. For example, the present invention can be applied to leafy vegetables such as lettuce, spinach, komatsuna, mizuna, cabbage, leaf radish, Chinese cabbage, and shiso; baby leaves such as romaine lettuce, beetroot, komatsuna, spinach, mizuna, arugula, mustard, kale, and chicory; medicinal herbs such as liquorice and ephedra; fruit vegetables such as tomatoes, eggplants, cucumbers, bell peppers, paprika, okra, chili peppers, pumpkins, strawberries, and blueberries; beans such as soybeans; and root vegetables such as leeks, onions, carrots, lotus roots, burdock, radishes, and potatoes.

The plant may be cultivated in any manner, i.e., planted in soil or cultivated submerged in a hydroponic solution. The plant functional component increaser of the present invention may be applied in any manner, for example, as a spray or immersion agent that is brought into contact with the stems, leaves, or roots of the plant, or as a soil irrigation agent. The present invention is highly advantageous because the functional components of the plant can be increased simply by spraying the plant functional component increaser of the present invention without the need for special equipment, etc.

The present invention also relates to plants with increased functional components cultivated by the above-mentioned cultivation method. Such plants are considered to be useful as food or as raw materials for cosmetics, medicines, supplements, etc.

The present invention will be described based on examples, but the present invention is not limited to only the examples.

Preparation of functional component increaser for test plant As a raw material, 2.8 g of linoleic acid with a purity of 80% (manufactured by Wako Pure Chemical Industries, Ltd.) was used, and 7 g of potassium carbonate (manufactured by Wako Pure Chemical Industries, Ltd.) and 300 ml of distilled water were added to prepare a reaction solution. The pH of the reaction solution at this time was 11.

0.2 mg of lipoxygenase (Sigma-Aldrich, derived from Glycine max) was added to the reaction solution and reacted at 30°C for 24 hours. The reaction mixture was then placed in a 90°C water bath for 5 minutes to inactivate the enzyme.

After the reaction solution in which the enzyme had been inactivated was returned to room temperature, 0.2 mg of alcohol dehydrogenase (Yeast-derived, manufactured by Wako Pure Chemical Industries, Ltd.) was added and the reaction was continued at 30°C for an additional 24 hours.

After the reaction was completed, the product in the reaction solution was identified by LC-MS using MS ² spectrum analysis with 13-oxo-9,11-octadecadienoic acid (manufactured by Cayman Chemical Company) as a standard substance, and quantified by the absolute calibration curve method at a detection wavelength of UV 272 nm.

13-oxo-9,11-octadecadienoic acid was obtained in a combined yield of 3.5% for isomers such as (E,E isomer) and (E,Z isomer). The yield (%) was calculated based on the following formula.
Yield (%) =
(wt% of 13-oxo-9,11-octadecadienoic acid produced)/(initial wt% of raw linoleic acid used)

The produced 13-oxo-9,11-octadecadienoic acid and its isomers were used to prepare an approximately 300 ppm potassium salt aqueous solution, which was used as an agent for increasing the functional components of the test plants and was evaluated as follows.

Effect of increasing functional components in strawberries: Example 1
Seven to nine strawberry plants (variety: Benihoppe) were cultivated in soil and irrigated with a general fertilizer solution. Just before flowering and fruiting began, the functional component increaser for the test plants was diluted 4000 times with water and irrigated at the base of the plants twice every six days at a rate of about 100 ml per plant. The fruits were harvested immediately after the irrigation, and 15 fruits were randomly selected from the harvested fruits for later analysis.
Comparative Example 1
The test was carried out in the same manner as in Example 1, except that the solution for irrigation was water instead of the functional component enhancer for the test plant.

Effect of increasing functional components in spinach: Example 2
About 50 spinach plants (variety: Benten) were grown in a greenhouse using soil cultivation. After the true leaves had developed, the functional component increaser for the test plant was diluted 4000 times with water and sprayed on the leaves at a rate of about 20 ml per plant. After treatment, the plants were grown for 15 days and harvested. Five plants were randomly selected from the harvested plants and used for subsequent analysis.
Comparative Example 2
The test was carried out in the same manner as in Example 2, except that the solution for irrigation was water instead of the functional component enhancer for the test plant.

Effect of increasing functional components in carrots: Example 3
Approximately 60 carrot plants (variety: Koyo No. 2) were cultivated in soil. After the primary leaves were developed, the functional component enhancer for the test plant was diluted 4000 times with water and sprayed on the leaves at a rate of about 50 ml per plant once every 7 days for 8 times. 60 days after the start of treatment, the carrot roots that had grown were collected. Six plants were randomly selected from the collected plants and used for subsequent analysis.
Comparative Example 3
The test was carried out in the same manner as in Example 3, except that the solution for irrigation was water instead of the functional component enhancer for the test plant.

Effect of increasing functional components in perilla: Example 4
About 20 plants of perilla (Oba; variety: Kaori Ao Oba) were cultivated in soil. After the true leaves were developed, the functional component enhancer for the test plant was diluted 4000 times with water and sprayed on the leaves at a rate of about 100 ml per plant once every 7 days, 4 times. 30 days after the start of treatment, the grown leaves were harvested, and 800 g of the harvested leaves were randomly selected and used for the subsequent analysis.
Comparative Example 4
The procedure was the same as in Example 4, except that the solution sprayed was water instead of the functional component enhancer for the test plant.

Effect of increasing functional components in eggplant: Example 5
Five eggplants (variety: Senryo 2) were cultivated in soil. After the primary leaves were developed, the functional component enhancer for the test plant was diluted 4000 times with water and the solution was irrigated at the base of the plant at a rate of about 100 ml per plant, once every seven days for four times. Thirty days after the start of the treatment, the grown fruit was harvested, and 2 kg of the harvested fruit was randomly selected and used for the subsequent analysis.
Comparative Example 5
The procedure was the same as in Example 5, except that the solution for irrigation was water instead of the functional component enhancer for the test plant.

Effect of increasing functional components in tomatoes: Example 6
Two cherry tomatoes were grown hydroponically in a greenhouse. Approximately two months after planting, 1-2 ml of the functional component enhancer for the test plants was added to the base of the plants once every seven days. Three to five cherry tomatoes were randomly selected from those harvested after treatment and analyzed, and the values over nine weeks were averaged to obtain the analytical value.
Comparative Example 6
The same procedure was followed as in Example 6, except that the solution added instead of the functional component enhancer for the test plant was water.

The amount and functionality of functional ingredients contained in the edible parts of each vegetable obtained in Examples 1-6 and Comparative Examples 1-6 (i.e., the fruit for strawberries, the above-ground parts for spinach, the roots for carrots, the leaves for shiso leaves, and the fruit parts for eggplant and tomato) were evaluated. Specifically, 1 g of strawberry sample was mashed in a blender and extracted with 10 ml of ethanol. 1 g of tomato sample was mashed in a blender and extracted with 7 ml of water (for GABA analysis), and the remaining residue was extracted with 8 ml of acetone (for lycopene analysis). The remaining vegetables were sent to Designer Foods Co., Ltd. and analysis was outsourced.

As for the functional components in the edible parts, functional components known to be generally contained in each vegetable were evaluated. Specifically, the total polyphenol content was measured by the Folin-Ciocalteu method in strawberries, lutein in spinach, β-carotene in carrots, lycopene and GABA in tomatoes were measured by HPLC, the total anthocyanin content was measured by UV-visible spectroscopy, and vitamin C was measured using an RQflex (registered trademark) (Merck) reflectometer. The results are shown in Table 1.

The antioxidant properties of each sample were evaluated as a functional index of the edible portion. Specifically, the antioxidant power of each sample was examined by measuring the superoxide scavenging ability, hydroxyl radical scavenging ability, and singlet oxygen scavenging ability of the sample against superoxide anion, hydroxyl radical, and singlet oxygen, which are representative active oxygen species, using an electron spin resonance (ESR) device, and the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging ability of the sample using absorptiometry (analysis was outsourced to Designer Foods Co., Ltd.). The results are shown in Table 2. In the table, the hydroxyl radical scavenging ability is the DMSO equivalent amount (μmol DMSO), the singlet oxygen scavenging ability is the His equivalent amount (μmol His), the DPPH radical scavenging ability is the Trolox equivalent amount (mg TE) using Trolox as the standard substance, and the value of the superoxide scavenging ability is the number of units of superoxide dismutase (SOD) equivalent to 1 g of sample.

As shown in Table 1, the amount of functional components contained in the vegetables of the example treated with the functional component increaser of the test plant was increased by 1.1 to 1.5 times compared to the comparative example. In addition, as shown in Table 2, the antioxidant properties of the vegetables of the example were also increased by about 1.1 to 1.4 times. For the perilla of Example 4, the antioxidant properties were not evaluated by measuring the antioxidant index. This is because, based on the experimental result that the amount of vitamin C, which is also used as a positive control in the evaluation of antioxidant function, increased by 30% in the perilla of Example 4 as shown in Table 1, it was presumed that the antioxidant properties of the perilla treated with the functional component increaser of the test plant were increased due to the increase in vitamin C. In addition, the antioxidant properties of the tomato of Example 6 were not evaluated. This is because, as shown in Table 1, it was considered self-evident that the antioxidant properties increased due to the increase in lycopene. Lycopene is widely recognized as having a powerful antioxidant effect far superior to that of vitamin E, a commonly known fat-soluble antioxidant, and is far superior to other carotenoids such as β-carotene and lutein. As shown in Table 1, another functional component that increased in tomatoes is GABA, a type of amino acid also known as γ-aminobutyric acid. Although it has a weak antioxidant effect, it is known to be a functional component that acts as an inhibitory neurotransmitter in the human body, and is used in many supplements and other nutritionally enhanced foods due to its relaxing and anti-stress effects. These results show that the plant functional component increaser of the present invention increases the amount of functional components contained in plants, and further, by increasing the amount of functional components with antioxidant activity, functional indicators such as antioxidant properties are clearly improved.

The above results show that the plant functional component increaser of the present invention has a significant effect of promoting the production of plant functional components and/or inhibiting the decomposition of plant functional components, and is an agent for increasing plant functional components that is excellent in increasing the functional components in the plant body.

Claims (8)

The formula:
HOOC-(R ¹ )-C=CC(=O)-R ² (I)
(Wherein,
R ¹ : a linear or branched alkylene group having 6 to 12 carbon atoms, ^which may contain a double bond that forms a conjugated double bond with the double bond between the α and β carbons of the carbonyl group in formula (I) ;
^R2 : an alkyl group having 2 to 8 carbon atoms.
An agent for increasing functional components of plants, characterized in that it contains an oxo fatty acid derivative or a salt thereof having the structural formula: The oxo fatty acid derivative,
The alkylene group of R ¹ has 8 to 10 carbon atoms,
2. The plant functional component enhancer according to claim 1 , wherein the alkyl group of ^R2 has 4 to 6 carbon atoms. The oxo fatty acid derivative,
R ¹ is an alkylene group having 9 carbon atoms;
3. The agent for increasing functional components in plants according to claim 2 , wherein ^R2 is an alkyl group having 5 carbon atoms. 4. The agent for increasing functional components in plants according to claim 3 , wherein the oxo fatty acid derivative is 13-oxo-9,11-octadecadienoic acid or a salt thereof. 5. The agent for increasing functional components of plants according to any one of claims 1 to 4 , which is used as a spray or immersion agent to be brought into contact with the stems, leaves or roots of plants, or as a soil drenching agent. The agent for increasing functional components of plants according to any one of claims 1 to 5 , wherein the functional components of plants are selected from the group consisting of vitamins, polyphenols, carotenoids, amino acids and combinations thereof. The plant functional component increaser according to any one of claims 1 to 5 , wherein the plant functional component is at least one selected from the group consisting of vitamin C, polyphenols, lutein, β-carotene, lycopene, and GABA. The agent for increasing functional components of plants according to any one of claims 1 to 7 , which is used for plants of the Solanaceae, Rosaceae, Apiaceae, Lamiaceae or Amaranthaceae family.