Plant growth promoter

Abstract

The present invention provides a method and feedstock for promoting the growth of leguminous plants comprised in legumes. Specifically disclosed is a growth promoter for leguminous plants contained in legumes, which is obtained by combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, the antioxidant (B) being at least one substance selected from the group consisting of tocopherol, dibutylated hydroxytoluene and butylated hydroxyanisole, the organic solvent (C) being one or more substances selected from the group consisting of an alcohol having 5 or less carbon atoms and dimethyl sulfoxide, and the organic solvent (C) being one or more substances selected from the group consisting of dimethyl sulfoxide and dimethyl sulfoxide. When the component (B) is set to be 1, the proportion (mass ratio) of the ascorbic acid or the salt thereof (A) is 10-3,000,000.

Description

Technical Field

[0001] The present invention relates to methods and materials for promoting the growth of legumes contained in beans. Background Art

[0002] About one-third of the land on Earth is dry, and it is expected that the number of dry areas will increase further due to global warming. In addition, as a countermeasure to the serious food shortage caused by population growth, it is urgent to develop technologies to improve, maintain, and increase the yield of grains such as soybeans, rice, wheat, and corn in dry areas, salt-intensive areas, high-temperature and low-temperature areas, that is, areas where growth is difficult or growth deteriorates and yields decrease. In particular, soybeans are an important grain and are widely consumed around the world, represented by Japan. And soybeans are different from other grains in that they have a high ratio of protein and lipids and rich nutritional value. Therefore, they are also very important as feed or oil raw materials, and technology development to increase yields is underway.

[0003] Oxygen is an indispensable substance for the survival of most animals and plants, but on the other hand, oxygen produces highly reactive active oxygen in animal and plant cells, causing serious damage to the organisms due to gene damage or enzyme inactivation. Therefore, a large amount of antioxidant substances including L-ascorbic acid are accumulated in plants, and a complex enzyme system that uses these antioxidant substances to remove active oxygen works in various parts of the cell.

[0004] From this point of view, raw materials applied to plants to promote growth are often combined with antioxidants including ascorbic acid. For example, non-patent document 1 discloses that the application of ascorbic acid to olives, sugar cane, wheat, etc. has achieved the effect of promoting growth and increasing yield. In addition, non-patent document 2 discloses that leguminous crops are sprayed with a certain concentration of ascorbic acid aqueous solution dissolved in distilled water on the leaves for a total of 3 times during their vegetative growth period or reproductive growth period, which can achieve an effect of increasing yield by up to 30%. In addition, patent document 1 discloses the use of an aqueous composition containing an iron (II) compound and L-ascorbic acid to promote plant growth, and patent document 2 discloses the use of an aqueous solution containing glycyrrhizic acid and L-ascorbic acid to promote plant growth.

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 60-202805

[0006] [Patent Document 2] Japanese Patent Application Laid-Open No. 08-143406

[0007] [Non-patent document 1] Akram et al. (2017) Ascorbic Acid-A Potential OxidantScavenger and Its Role in Plant Development and Abiotic StressTolerance. Frontiers in Plant Science, 8: 613

[0008] [Non-patent document 2] Zarghamnejad et al., (2014) Chickpea response to ascorbicacid foliar application at vegetative and reproductive stages. International Journal of Biosciences, 5: 166-170 Summary of the invention

[0009] The present invention relates to the following 1) to 15).

[0010] 1) A growth promoter for leguminous plants contained in beans, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0011] 2) A growth promoter for leguminous plants contained in beans, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0012] 3) A nodule activity promoter, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0013] 4) A nodule activity promoter, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0014] 5) A yield-increasing agent for leguminous plants contained in beans, wherein the agent is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0015] 6) A yield-increasing agent for leguminous plants contained in beans, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0016] 7) A method for promoting the growth of legumes contained in beans, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0017] 8) A method for promoting root nodule activity, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0018] 9) A method for increasing the yield of legumes contained in beans, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0019] 10) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a growth promoter for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0020] 11) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a growth promoter for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0021] 12) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the manufacture of a nodule activity promoter, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the proportion (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0022] 13) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the manufacture of a nodule activity promoter, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0023] 14) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a yield-increasing agent for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0024] 15) Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a yield-increasing agent for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is the seed weight of each soybean individual.

[0026] Figure 2 The solubility when the mixing ratio of BHT and isobutyl alcohol is changed.

[0027] Figure 3 It is the solubility of a mixture of BHT and an organic solvent (1:4).

[0028] Figure 4 It is the weight of soybean nodules one week after foliar spraying.

[0029] Figure 5 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0030] Figure 6 It is the amount of allantoin contained in soybean exudate one week after foliar spraying.

[0031] Figure 7 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0032] Figure 8 This is the soybean plant body in test plot 7 one week after foliar spraying.

[0033] Fig. 9 It is the seed weight of each soybean individual.

[0034] Fig.10 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0035] Fig.11 It is the dry weight of the aboveground part of red beans one week after foliar spraying.

[0036] Fig.12 It is the dry weight of the underground part of red beans one week after foliar spraying.

[0037] Fig.13 It is the dry weight of the aboveground part of chickpeas one week after foliar spraying.

[0038] Fig.14 It is the number of pods per individual Lotus japonicus.

[0039] Fig.15 It is the seed weight of each individual of Lotus japonicus.

[0040] Fig.16 It is the number of beans per individual broad bean.

[0041] Fig.17 It is the weight of the grains of each broad bean individual.

[0042] Fig.18 This is the appearance of a single-dose or double-dose composition after storage for 1 week.

[0043] Fig.19 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0044] Fig. 20 It is the dry weight of the aboveground part of soybeans one week after foliar spraying.

[0045] Fig.21 It is the dry weight of the underground part of soybeans one week after foliar spraying. DETAILED DESCRIPTION

[0046] The present invention provides methods and materials for promoting the growth of legumes contained in beans.

[0047] The inventors of the present invention have studied the use of ascorbic acid to promote plant growth and have found that when an ascorbic acid solution dissolved in water used in agricultural fields such as tap water or well water is sprayed on leguminous plants including beans, the growth promoting effect of ascorbic acid is hardly obtained.

[0048] Furthermore, the inventors have found that the combination of ascorbic acid and a specific antioxidant dissolved in an organic solvent can exert a nodule activity promoting effect and a growth promoting effect.

[0049] According to the method of the present invention, the growth amount of the legume contained in the beans can be increased. In addition, the nodule activity of the legume contained in the beans with nodules formed is promoted, and the urea production ability of the plant is improved. That is, according to the present invention, the yield of fruits, seeds, or grains of the legume contained in the beans can be increased.

[0050] In the growth promoter of the legume contained in beans of the present invention, "growth promotion" means increasing the growth amount (fresh weight, elongation, etc.) of the legume contained in beans, and increasing the yield of fruits, seeds, or grains. When the growth promoter of the legume contained in beans of the present invention is specifically intended to increase the grain yield, it is referred to as a "yield-increasing agent for legumes contained in beans".

[0051] In the present invention, the Fabaceae plants included in the beans refer to so-called "beans" harvested from seeds of soybeans (including edamame), red beans, chickpeas, lotus roots, kidney beans, peanuts, broad beans, peas, runner beans, lima beans, mung beans, cowpeas, black-eyed peas, lentils, sword beans, lentils, and winged beans. The Fabaceae plants included in the beans are preferably soybeans, red beans, kidney beans, peas, broad beans, chickpeas, and lotus roots, and more preferably soybeans.

[0052] In the nodule activity promoter of the present invention, "nodule activity promotion" means promotion of nodule activity in nodule-forming plants, that is, promotion of the nitrogen fixation function in the host plant performed by nodules.

[0053] Here, "root nodule" refers to a nodule that grows on the root system of a plant due to symbiosis with bacteria (rhizobium). Rhizobium reduces atmospheric nitrogen to ammonia nitrogen in the nodule and provides it to the host in so-called symbiotic nitrogen fixation.

[0054] The "nodule-forming plant" refers to a host plant that grows nodules, and in the present invention, preferably, leguminous plants are included in beans.

[0055] The growth promoter or nodule activity promoter of leguminous plants contained in beans of the present invention is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent.

[0056] The combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent is prepared by premixing the components (A), (B) and (C) at an appropriate mass ratio or mixing them at the time of use. As one embodiment, components (A), (B) and (C) can be combined to form a single preparation (single-dose preparation (composition)); as another embodiment, a preparation containing component (A) and a preparation containing components (B) and (C) are separately prepared, and then a two-dose preparation (set) is prepared in which the two are combined at the time of use.

[0057] In the present invention, the IUPAC systematic name of the component (A) ascorbic acid is (R)-3,4-dihydroxy-5-((S)-1,2-dihydroxyethyl)furan-2(5H)-one. Ascorbic acid may be any of the D-form, L-form and DL-form, but is preferably the L-form (so-called L-ascorbic acid).

[0058] As ascorbic acid, commercially available products of various grades can be used.

[0059] Examples of the salt of ascorbic acid include sodium salts, potassium salts, calcium salts, magnesium salts, barium salts, ammonium salts, and salts with nitrogen-containing organic bases such as pyridine, trimethylamine, triethylamine, tributylamine, and diethylamine.

[0060] When the growth promoter or nodule activity promoter of a legume plant contained in beans of the present invention is applied to plants, the concentration of (A) ascorbic acid or a salt thereof in the composition can be appropriately adjusted in the range of 100 mass ppm to 300,000 mass ppm, for example, depending on the method of administration.

[0061] For example, when spraying is performed using a sprayer (e.g., a plunger pump sprayer), the amount of the compound in the spray liquid is preferably 100 mass ppm or more, more preferably 300 mass ppm or more, and even more preferably 500 mass ppm or more, and is preferably 20,000 mass ppm or less, more preferably 10,000 mass ppm or less, and even more preferably 4,500 mass ppm or less. In addition, it is preferably 100 to 20,000 mass ppm, more preferably 300 to 10,000 mass ppm, and even more preferably 500 to 4,500 mass ppm.

[0062] In the case of aerial spraying, the amount of the spray liquid is preferably 20,000 mass ppm or more, more preferably 80,000 mass ppm or more, and further preferably 150,000 mass ppm or more, and is preferably 300,000 mass ppm or less, more preferably 250,000 mass ppm or less, and further preferably 200,000 mass ppm or less. In addition, it is preferably 20,000 to 300,000 mass ppm, more preferably 80,000 to 250,000 mass ppm, and further preferably 150,000 to 200,000 mass ppm.

[0063] In the present invention, the antioxidant component (B) is specifically at least one selected from tocopherol (vitamin E), butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA). Among them, BHT and BHA, which are phenolic antioxidants, are preferred, and BHT is more preferred.

[0064] In addition, in the present invention, as an antioxidant, in addition to antioxidants other than tocopherol, BHT and BHA, at least one selected from, for example, sodium isoascorbate, propyl gallate, sodium sulfite, potassium sulfite, potassium pyrosulfite, chlorogenic acid, catechin, glutathione, uric acid, etc. can be used in combination, and preferably at least one selected from sodium sulfite, potassium sulfite, potassium pyrosulfite, chlorogenic acid, catechin, glutathione and uric acid can be used in combination.

[0065] Therefore, as component (B) antioxidant, as a preferred embodiment, one or more phenolic antioxidants selected from BHT and BHA are used, and in addition to the phenolic antioxidant, one or more selected from sodium sulfite, potassium sulfite, glutathione and uric acid are used in combination.

[0066] When ascorbic acid is dissolved in water containing metal ions such as tap water, the metal ions react with ascorbic acid to generate hydrogen peroxide, and the hydrogen peroxide further reacts with the metal ions to generate hydroxyl radicals. Thus, once hydroxyl radicals invade the plant body, they will trigger a continuous lipid peroxidation reaction that reacts with lipids present in the cell membrane, etc., producing lipid free radicals or lipid peroxyl radicals. By using antioxidants, such free radicals can be captured.

[0067] In the present invention, the (B) antioxidant is used in a solution state dissolved in the (C) organic solvent, whereby the plant growth promoting or nodule promoting effect of ascorbic acid can be effectively exerted.

[0068] The growth promoter or nodule activity promoter of leguminous plants contained in beans of the present invention may be a single-dose preparation or a two-dose preparation as described above, but may also be used in a manner such that a preparation containing component (A) (first preparation) and a preparation containing components (B) and (C) (second preparation) are separately prepared, and then these are combined into a two-dose preparation (set), and the two are mixed when used. By making it into a two-dose preparation, it is possible to suppress the discoloration of the preparation containing all of components (A), (B) and (C) during storage.

[0069] In this case, the first agent containing component (A) may contain an antioxidant other than tocopherol, BHT and BHA, for example, one or more selected from the group consisting of sodium sulfite, potassium sulfite, potassium pyrosulfite, chlorogenic acid, catechin, glutathione and uric acid.

[0070] When the growth promoter or nodule activity promoter of a leguminous plant contained in beans of the present invention is applied to plants, the concentration of the antioxidant (B) in the composition is preferably 0.001 mass ppm or more, more preferably 0.01 mass ppm or more, and even more preferably 0.1 mass ppm or more, and is preferably 100 mass ppm or less, more preferably 20 mass ppm or less, and even more preferably 5 mass ppm or less. In addition, it is preferably 0.001 to 100 mass ppm, more preferably 0.01 to 20 mass ppm, and even more preferably 0.1 to 5 mass ppm.

[0071] The organic solvent as the component (C) is a solvent for dissolving the antioxidant (B), and specifically is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide (DMSO).

[0072] Here, preferred examples of the alcohol having 5 or less carbon atoms include ethanol, methanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, 2-methyl-2-propanol, isobutanol, 1-pentanol, 3-methyl-1-butanol, 2,2-dimethyl-1-propanol, ethylene glycol, diethylene glycol, propylene glycol, and glycerol, with isobutanol and DMSO being more preferred.

[0073] When one or more selected from BHT and BHA is used as the (B) antioxidant, DMSO, isobutyl alcohol or a mixture thereof is preferably used as the (C) organic solvent.

[0074] When the growth promoter or nodule activity promoter of leguminous plants contained in beans of the present invention is applied to plants, the concentration of the organic solvent (C) in the composition can be appropriately adjusted within the range of 1 ppm by mass to 100,000 ppm by mass depending on the method of application.

[0075] For example, when spraying is performed using a sprayer (e.g., a plunger pump sprayer), the amount of the compound in the spray liquid is preferably 1 mass ppm or more, more preferably 10 mass ppm or more, and further preferably 50 mass ppm or more, and is preferably 10,000 mass ppm or less, more preferably 5,000 mass ppm or less, and further preferably 1,000 mass ppm or less. In addition, it is preferably 1 to 10,000 mass ppm, more preferably 10 to 5,000 mass ppm, and further preferably 50 to 1,000 mass ppm.

[0076] In the case of aerial spraying, the amount of the pesticide in the spray liquid is preferably 100 mass ppm or more, more preferably 1,000 mass ppm or more, and even more preferably 5,000 mass ppm or more, and is preferably 100,000 mass ppm or less, more preferably 50,000 mass ppm or less, and even more preferably 10,000 mass ppm or less. In addition, it is preferably 100 to 100,000 mass ppm, more preferably 1,000 to 50,000 mass ppm, and even more preferably 5,000 to 10,000 mass ppm.

[0077] As the ratio (mass ratio) of the combination of (A) ascorbic acid or a salt thereof, (B) antioxidant and (C) organic solvent, when component (B) is set to 1, component (A) is preferably 10 or more, more preferably 100 or more, and further preferably 500 or more, and is preferably 3,000,000 or less, more preferably 2,000,000 or less, and further preferably 200,000 or less. In addition, it is preferably 10 to 3,000,000, more preferably 100 to 2,000,000, and further preferably 500 to 200,000. Among them, when spraying is performed by a sprayer, it is preferably 500 to 45,000; when spraying is performed by aerial spraying, it is preferably 150,000 to 2,000,000. Furthermore, when spraying is performed by a sprayer, it is more preferably 500 to 4,500, and when spraying is performed by aerial spraying, it is more preferably 150,000 to 200,000.

[0078] In addition, when component (B) is 1, component (C) is preferably 4 or more, more preferably 10 or more, further preferably 20 or more, and further preferably 50 or more, and is preferably 100,000 or less, more preferably 50,000 or less, further preferably 20,000 or less, and further preferably 10,000 or less. In addition, it is preferably 4 to 100,000, more preferably 10 to 50,000, further preferably 20 to 20,000, and further preferably 50 to 10,000. Among them, when spraying by a sprayer, it is preferably 20 to 10,000; when spraying by aerial spraying, it is preferably 2,000 to 100,000. Furthermore, when spraying by a sprayer, it is more preferably 50 to 1,000; when spraying by aerial spraying, it is more preferably 5,000 to 10,000.

[0079] In the present invention, (D) a surfactant can be used in combination with (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent.

[0080] By using a surfactant, the wettability, adhesion and permeability of (A) ascorbic acid or a salt thereof to the plant surface can be improved, so that the effect of (A) ascorbic acid or a salt thereof can be enhanced or the effect can be more efficiently exerted.

[0081] When the growth promoter or nodule activity promoter for legumes contained in beans of the present invention is prepared as a two-dose preparation (set) comprising a preparation (first agent) containing component (A) and a preparation (second agent) containing components (B) and (C) as described above, the surfactant (D) is preferably contained in the second agent containing components (B) and (C).

[0082] As the surfactant, a nonionic surfactant and / or anionic surfactant may be contained.

[0083] Examples of the nonionic surfactant include, for example, one or more selected from sorbitan fatty acid esters, glycerol fatty acid esters, polyoxyalkylene sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyglycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters, polyoxyethylene alkyl ethers, polyoxyethylene alkyl aryl ethers, polyoxyethylene aryl ethers, polyoxyethylene alkenyl ethers, alkyl glycosides, polyoxyalkylene alkyl glycosides, and sucrose fatty acid esters. Examples of the anionic surfactant include, for example, one or more selected from alkyl sulfates, alkylbenzene sulfonates, polyoxyalkylene alkyl ether sulfates, polyoxyalkylene alkenyl ether sulfates, and polyoxyalkylene alkyl aryl ether sulfates, fatty acid salts, pyrophosphates, lauryl phosphoric acid, polycarboxylic acid type polymers, polyoxyethylene alkylene alkyl acetates, aromatic sulfonate formaldehyde condensates, polyoxyethylene distyryl ether sulfates, alkyl diphenyl ether disulfonates, dialkyl sulfosuccinates, alkyl naphthalene sulfonates, and the like.

[0084] Among them, from the viewpoint of preventing the phytotoxicity to plants during excessive application, as nonionic surfactant, it is preferably selected from one or more of sorbitan fatty acid ester, glycerol fatty acid ester, polyoxyalkylene sorbitan fatty acid ester, polyalkylene glycol fatty acid ester, polyoxyethylene resin acid ester, polyoxyethylene alkyl ether, alkyl glycoside and sucrose fatty acid ester, more preferably selected from one or more of sorbitan fatty acid ester, glycerol fatty acid ester, polyalkylene glycol fatty acid ester, polyoxyethylene resin acid ester and polyoxyethylene alkyl ether. As anionic surfactant, it is preferably selected from one or more of alkyl sulfate (e.g. sodium lauryl sulfate, ammonium lauryl sulfate, triethanolamine lauryl sulfate, etc.) and fatty acid salt.

[0085] When the growth promoter or nodule activity promoter of leguminous plants contained in beans of the present invention is applied to plants, the concentration of the surfactant (D) in the composition can be appropriately adjusted within the range of 10 to 30,000 ppm by mass depending on the method of administration.

[0086] For example, when spraying is performed using a sprayer (e.g., a plunger pump sprayer), the amount of the compound in the spray liquid is preferably 10 mass ppm or more, more preferably 100 mass ppm or more, and even more preferably 200 mass ppm or more, and is preferably 5,000 mass ppm or less, more preferably 1,000 mass ppm or less, and even more preferably 500 mass ppm or less. In addition, it is preferably 10 to 5,000 mass ppm, more preferably 10 to 1,000 mass ppm, and even more preferably 100 to 500 mass ppm.

[0087] In the case of aerial spraying, the amount of the spray liquid is preferably 100 ppm by mass or more, more preferably 500 ppm by mass or more, and even more preferably 1,000 ppm by mass or more, and is preferably 10,000 ppm by mass or less, more preferably 8,000 ppm by mass or less, and even more preferably 5,000 ppm by mass or less. In addition, it is preferably 100 to 10,000 ppm by mass, more preferably 500 to 8,000 ppm by mass, and even more preferably 1,000 to 5,000 ppm by mass.

[0088] In addition, as a ratio (mass ratio) when the surfactant (D) is combined, when the component (B) is set to 1, the component (D) is preferably 10 or more, more preferably 50 or more, and further preferably 100 or more, and is preferably 300,000 or less, more preferably 100,000 or less, and further preferably 5,000 or less. In addition, it is preferably 10 to 300,000, more preferably 50 to 100,000, and further preferably 100 to 5,000. Among them, when spraying with a sprayer, it is preferably 100 to 5,000; when spraying in the air, it is preferably 1,000 to 50,000. Furthermore, when spraying with a sprayer, it is more preferably 100 to 500; when spraying in the air, it is more preferably 1,000 to 5,000.

[0089] Furthermore, in the present invention, (E) a chelating agent can be used in combination with (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent.

[0090] By using a chelating agent, the stability of (A) ascorbic acid or a salt thereof can be improved, and as a result, the effect of ascorbic acid or a salt thereof can be stabilized.

[0091] When the growth promoter or nodule activity promoter for leguminous plants contained in beans of the present invention is prepared as a two-dose preparation (set) comprising a preparation (first preparation) containing the ingredient (A) and a preparation (second preparation) containing the ingredients (B) and (C) as described above, the (E) chelating agent is preferably contained in the first preparation containing the ingredient (A).

[0092] As chelating agents, for example, aminocarboxylic acid chelating agents, phosphonic acid chelating agents, hydroxycarboxylic acid chelating agents, polycarboxylic acid chelating agents, etc. can be cited. As aminocarboxylic acid chelating agents, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), methylglycinediacetic acid (MGDA), triethylenetetraaminehexaacetic acid (TTHA), glutamic aciddiacetic acid (GLDA), hydroxyethyliminediacetic acid (HIDA), dihydroxyethylglycine (DHEG), aspartic aciddiacetic acid (ASDA), ethylenediaminesuccinic acid (EDDS) and salts thereof can be cited. As phosphonic acid chelating agents, hydroxyethylidene diphosphonic acid (HEDP), nitrilotrimethylenephosphonic acid (NTMP), phosphinobutanetricarboxylic acid (PBTC), ethylenediaminetetramethylenephosphonic acid (EDTMP) and salts thereof can be cited.

[0093] Examples of the hydroxycarboxylic acid chelating agent include citric acid, malic acid, tartaric acid, gluconic acid, lactic acid, and salts thereof.

[0094] Examples of the polycarboxylic acid chelating agent include succinic acid, oxalic acid, glutaric acid, adipic acid, fumaric acid, malonic acid, and salts thereof.

[0095] When the growth promoter or nodule activity promoter of a leguminous plant contained in beans of the present invention is applied to a plant, the concentration of the chelating agent (E) in the composition is preferably 0.01 mass ppm or more, more preferably 0.1 mass ppm or more, and further preferably 1 mass ppm or more, and is preferably 100 mass ppm or less, more preferably 50 mass ppm or less, and further preferably 10 mass ppm or less. In addition, it is preferably 0.01 to 100 mass ppm, more preferably 0.1 to 50 mass ppm, and further preferably 1 to 10 mass ppm.

[0096] In addition, as a ratio (mass ratio) when the chelating agent (E) is combined, when the component (B) is set to 1, the component (E) is preferably 0.01 or more, more preferably 0.1 or more, and further preferably 1 or more, and is preferably 100 or less, more preferably 50 or less, and further preferably 10 or less. In addition, it is preferably 0.01 to 100, more preferably 0.1 to 100, more preferably 1 to 100, and further preferably 1 to 10.

[0097] As a preferred combination of (B) an antioxidant, (C) an organic solvent, and (D) a surfactant used together with (A) ascorbic acid, for example, the following combinations can be mentioned.

[0098] A combination comprising a component (B), a component (C), a component (D) and, if necessary, a surfactant further comprising an alkyl sulfate, wherein the component (B) is an antioxidant comprising at least one selected from the group consisting of BHT and BHA, the component (C) is an organic solvent comprising isobutyl alcohol or DMSO, and the component (D) is at least one selected from the group consisting of sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters and polyoxyethylene alkyl ethers.

[0099] A combination comprising a component (B), a component (C), a component (D) and, if necessary, a surfactant further comprising an alkyl sulfate, wherein the component (B) is an antioxidant comprising BHT, sodium sulfite, glutathione and uric acid, the component (C) is an organic solvent comprising isobutyl alcohol or DMSO, and the component (D) is one or more selected from the group consisting of sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters and polyoxyethylene alkyl ethers.

[0100] As shown in the examples described below, when soybean seeds, which are nodule-forming plants, are sown and then the soybeans are inoculated with rhizobia and cultivated, if an antioxidant such as BHT and an organic solvent are added in combination with ascorbic acid, the nodule weight and nodule activity increase, and urea production by the nodules is promoted, indicating that the growth of the host plant is promoted.

[0101] Therefore, the combination of ascorbic acid, a specific antioxidant and an organic solvent can be a growth promoter or a nodule activity promoter for legumes contained in beans, can be used for growth promotion or nodule activity promotion of legumes contained in beans, and can be used to manufacture a growth promoter or a nodule activity promoter for legumes contained in beans. It has been reported that the amount of urea generated from nodules is correlated with grain yield (rotation field soybean total base fertilizer nitrogen diagnosis method Ibaraki Prefecture Agricultural Comprehensive Center-Agricultural Research Institute H18 main results, http: / / www.pref.ibaraki.jp / nourinsuisan / noken / seika / h18pdf / documents / 27.pdf), so it can be considered that such a nodule activity promoter is effective for increasing grain yield.

[0102] The nitrogen fixation function of the host plant can be evaluated by urea production ability or amide production ability. Therefore, the nodule activity of the present invention can be specifically evaluated by the urea production amount or amide production amount per unit wet nodule weight. Such urea production amount or amide production amount can be calculated by measuring the urea production amount or amide amount in the exudate collected when the plant body is cut (for example, the aerial part is cut off just below the cotyledon). Here, as urea, allantoin, allantoic acid or citrulline can be listed. As amide, asparagine or glutamine can be listed. Preferably, the amount of allantoic acid or asparagine is measured.

[0103] The above-mentioned growth promoter or nodule activity promoter of legumes contained in beans can be used as a composition for promoting the growth or nodule activity of legumes contained in beans (such as various agricultural or horticultural materials, etc.), or a raw material (monomer) or preparation for adding or mixing in a cultivation base material for cultivating plants such as soil, culture medium, and soilless culture solution.

[0104] The composition may be in the form of a liquid or gel composition, or may be in the form of a solid state (block, powder, granular, etc.).

[0105] Here, the composition may be a composition obtained by premixing components (A), (B) and (C), or a composition obtained by separately preparing a preparation containing component (A) and a preparation containing components (B) and (C) and mixing the two when used.

[0106] In addition to the above-mentioned components (A) to (C), and (D) and (E) used in the present invention, the above-mentioned composition may also contain optional components. Examples of such components include solvents (e.g., water, buffer, culture medium, soilless culture solution, etc.), carriers (e.g., zeolite, silica, bentonite, sodium sulfate, diatomaceous earth, vermiculite, perlite, peat moss, activated carbon, humus, talc, clay, carbon black, pulp, straw, bean curd, kaolin, montmorillonite, alumina, etc.), pH regulators for promoting dissolution of the above-mentioned compounds, spreading agents for improving the spreading power on plants or soil, fertilizer components for improving fertilizer efficiency, pesticide components, adhesives, extenders, Plant growth promoting microorganisms such as rhizobia or mycorrhizal fungi, essential nutrients for plants, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleobases, sugars, monohydric alcohols, food additives, microbial extracts, plant hormones, nod factors, i.e. lipid chito-oligosaccharides, synthetic lipid chito-oligosaccharides, chito-oligosaccharides, chitin compounds, linoleic acid or its derivatives, linolenic acid or its derivatives, karrikin, acyl-homoserine lactone derivatives, betaine compounds, phenolic compounds, etc.

[0107] When the growth promoter or nodule activity promoter of leguminous plants contained in beans of the present invention is used in the form of a single-dose preparation, it is preferred to add one or more carriers selected from zeolite, silica, bentonite and sodium sulfate from the viewpoint of storage stability or inhibition of browning of the preparation, and silica is more preferred from the viewpoint of storage stability and inhibition of browning. In this case, the concentration of the carrier (component (F)) in the composition is preferably 50 mass ppm or more, more preferably 100 mass ppm or more, and further preferably 250 mass ppm or more, and is preferably 1,000 mass ppm or less, more preferably 750 mass ppm or less, and further preferably 650 mass ppm or less. In addition, it is preferably 50 to 1,000 mass ppm, more preferably 100 to 750 mass ppm, and further preferably 250 to 650 mass ppm.

[0108] In addition, as a ratio (mass ratio) when the (F) carrier is combined, when the component (B) is set to 1, the component (F) is preferably 50 or more, more preferably 100 or more, and further preferably 250 or more, and is preferably 1,000 or less, more preferably 750 or less, and further preferably 650 or less. In addition, it is preferably 50 to 1,000, more preferably 100 to 750, and further preferably 250 to 650.

[0109] The composition for aerial spraying may contain an emulsifier such as polyoxyethylene fatty acid ester or an oil such as decanol.

[0110] Examples of the above-mentioned composition include, but are not limited to, a cultivation base material containing at least the components (A) to (C) of the present invention (e.g., agricultural or horticultural soil, culture soil, culture medium, soilless culture solution, water, etc.), fertilizers, irrigation water, microbial materials such as rhizobia materials, soil conditioners, pesticides, materials for sowing, plant supplements (e.g., activators, nutrients, etc.), etc.

[0111] The fertilizer, microbial material, soil conditioner, material for sowing, and plant supplement are preferred because they help improve the soil for growing plants. The fertilizer, microbial material, soil conditioner, material for sowing, and plant supplement may be solid or liquid, and in the case of solid, may be in the form of blocks, powders, granules, etc., preferably powders or granules. In addition to containing components (A) to (C) as active ingredients, the fertilizer, microbial material, soil conditioner, material for sowing, and plant supplement may also contain components of fertilizers, microbial materials, soil conditioners, material for sowing, and plant supplements that are commonly used for plant cultivation.

[0112] The cultivation substrate, fertilizer, microbial material such as rhizobium material, soil conditioner, pesticide, material for sowing, plant supplement can be prepared by adding components (A) to (C) of the present invention to a common cultivation substrate (e.g., agricultural or horticultural soil, culture soil, culture medium, soilless culture solution, water, etc.), fertilizer, microbial material such as rhizobium material, soil conditioner, pesticide, material for sowing, plant supplement (e.g., activator, nutrient, etc.), etc.

[0113] The composition may be prepared separately from the preparation containing component (A) and the preparation containing components (B) and (C) and mixed them before use. In the case of a two-dose preparation in which a preparation containing component (A) (first dose) and a preparation containing components (B) and (C) (second dose) are prepared and mixed before use, the weight ratio of the first dose is 1 to 100, preferably 1 to 50, and more preferably 1 to 25, when the second dose is taken as 1.

[0114] When the first agent and the second agent are prepared by dissolving them separately in water, the concentration of the first agent in the aqueous solution is preferably 100 mass ppm to 300,000 mass ppm, and the concentration of the second agent in the aqueous solution is preferably 10 mass ppm to 100,000 mass ppm.

[0115] The method for supplying the growth promoter or nodule activity promoter for leguminous plants contained in beans of the present invention is not particularly limited as long as the agent can be applied to plants to exhibit the effects of the present invention.

[0116] That is, as long as the composition contacts or reaches the soil of the plant body or plant root system of the legume plant included in the beans, there is no particular limitation, and examples include spraying on the soil surface, pouring, hoeing, spraying on the leaves of plants, mixing in fertilizers, adding to hydroponic solutions, or applying or smearing on seeds before sowing (e.g., seed coating), etc., but it is preferably applied in the form of a spray liquid in which the components of the present invention are diluted with water, and foliar spraying is particularly suitable.

[0117] The spray liquid can be prepared at the time of application, and the dilution water used at this time may be any of agricultural water, well water, groundwater, river water, lake water, tap water, and the like.

[0118] The method of spraying is not particularly limited, and examples thereof include a spraying method, that is, a method of spraying a spray liquid in a mist form by spraying. By such a method, the leguminous plant growth promoter or nodule activity promoter contained in beans of the present invention can exhibit good spreadability on the plant after being attached to the plant.

[0119] Specific examples of the spraying method include manual spraying using a watering can, atomizer, sprayer (for example, a plunger pump sprayer), and the like, and aerial spraying using an airplane, a helicopter, an unmanned aerial vehicle, and the like.

[0120] The amount of the legume growth promoter or nodule activity promoter contained in beans of the present invention to be applied depends on the concentration of components (A) to (C) contained in the composition at the time of application. For example, when the concentration of component (A) contained in the spray liquid is 100 to 300,000 mass ppm, the amount of component (A) used per plant in the composition is preferably 1 mg or more, more preferably 5 mg or more, and further preferably 10 mg or more, and preferably 150 mg or less, more preferably 100 mg or less, and further preferably 50 mg or less. In addition, it is preferably 1 to 150 mg, more preferably 5 to 100 mg, and further preferably 10 to 50 mg. The legume growth promoter or nodule activity promoter contained in beans can be applied in the above range at one time or in multiple applications.

[0121] The timing and frequency of application vary depending on the type of legume included in the beans. In the case of soybeans, it is usually applied by spraying, pouring, hoeing or seed coating on the surface of a cultivation substrate such as soil, preferably once or 1 to 3 times before or at the same time as sowing. In the case of application after sowing, it is preferably between the early stage of the vegetative growth period before entering the reproductive growth period and the grain swelling period after entering the reproductive growth period.

[0122] Regarding the above-mentioned embodiment, the present invention also discloses the following aspects.

[0123] <1> A growth promoter for leguminous plants contained in beans, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0124] <2> A growth promoter for leguminous plants contained in beans, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0125] <3> A nodule activity promoter, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0126] <4> A nodule activity promoter, wherein the preparation is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0127] <5> A yield-increasing agent for leguminous plants contained in beans, wherein the agent is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0128] <6> A yield-increasing agent for leguminous plants contained in beans, wherein the agent is a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is one or more selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0129] <7> The preparation according to any one of <1> to <6>, wherein, with respect to the ratio (mass ratio) of (A) ascorbic acid or a salt thereof to (C) an organic solvent, when component (B) is set to 1, component (A) is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, and component (C) is 4 to 100,000, preferably 10 to 50,000, more preferably 20 to 20,000, and even more preferably 50 to 10,000.

[0130] <8> The preparation according to any one of <1> to <7>, wherein the (B) antioxidant further comprises one or more selected from the group consisting of sodium sulfite, potassium sulfite, glutathione and uric acid.

[0131] <9> The preparation according to any one of <1> to <8>, wherein the (C) organic solvent is at least one selected from isobutyl alcohol and dimethyl sulfoxide.

[0132] <10> The preparation according to any one of <1> to <9>, further comprising (D) a surfactant.

[0133] <11> In the preparation according to <10>, when the component (B) is 1, the ratio (mass ratio) of the surfactant (D) is preferably 10 to 300,000, more preferably 50 to 100,000, and even more preferably 100 to 5,000.

[0134] <12> The preparation according to <10> or <11>, wherein the (D) surfactant comprises a nonionic surfactant and / or an anionic surfactant.

[0135] <13> The preparation according to <12>, wherein the (D) nonionic surfactant is at least one selected from the group consisting of sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters and polyoxyethylene alkyl ethers, and the anionic surfactant is an alkyl sulfate ester salt.

[0136] <14> The preparation according to <12>, wherein the (D) surfactant comprises a nonionic surfactant and an anionic surfactant.

[0137] <15> A preparation as described in <14>, wherein (D) the nonionic surfactant is at least one selected from sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters and polyoxyethylene alkyl ethers, and the anionic surfactant is at least one selected from alkyl sulfate ester salts and fatty acid salts.

[0138] <16> The preparation according to any one of <1> to <15>, further comprising (E) a chelating agent.

[0139] <17> The preparation according to <16>, wherein, with respect to the ratio (mass ratio) of (A) ascorbic acid or a salt thereof, (C) an organic solvent, (D) a surfactant and (E) a chelating agent, when component (B) is taken as 1, component (A) is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, component (C) is 4 to 100,000, preferably 10 to 50,000, more preferably 20 to 20,000, further preferably 50 to 10,000, component (D) is preferably 10 to 300,000, more preferably 50 to 100,000, further preferably 100 to 5,000, and component (E) is preferably 0.01 to 100, more preferably 0.1 to 100, more preferably 1 to 100, further preferably 1 to 10.

[0140] <18> The preparation according to <16> or <17>, wherein the (E) chelating agent is at least one selected from ethylenediaminetetraacetic acid and ethylenediamine-N,N′-disuccinic acid.

[0141] <19> The preparation according to any one of <1> to <18>, further comprising (F) a carrier.

[0142] <20> The preparation according to <19>, wherein the ratio (mass ratio) of the carrier (F) is 10 to 10,000, preferably 50 to 1,000, more preferably 100 to 750, and even more preferably 250 to 650, relative to component (B).

[0143] <21> The preparation according to <19> or <20>, wherein the carrier (F) is at least one selected from the group consisting of zeolite, silica, bentonite and sodium sulfate.

[0144] <22> The preparation according to any one of <1> to <9>, comprising a first agent containing the above-mentioned component (A) and a second agent containing the above-mentioned components (B) and (C), and the two agents are combined when used.

[0145] <23> A method for promoting the growth of legumes contained in beans, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0146] <24> A method for promoting root nodule activity, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0147] <25> A method for increasing the yield of legumes contained in beans, comprising the step of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the mixture to soil or plants, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when the component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0148] <26> The method according to any one of <23> to <25>, wherein a first agent containing the component (A) and a second agent containing the components (B) and (C) are prepared, and when used, the two agents are applied in combination to soil or plants.

[0149] <27> The preparation according to <22>, wherein the weight ratio of the first dose is 1 to 100, when the second dose is assumed to be 1.

[0150] <28> The method according to <26>, wherein the first agent and the second agent are dissolved in water before use.

[0151] <29> The method according to <28>, wherein the concentration of the first agent in the aqueous solution is from 100 mass ppm to 300,000 mass ppm, and the concentration of the second agent is from 10 mass ppm to 100,000 mass ppm.

[0152] <30> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a growth promoter for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0153] <31> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the production of a growth promoter for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when the component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0154] <32> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the manufacture of a nodule activity promoter, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the proportion (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0155] <33> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the manufacture of a nodule activity promoter, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0156] <34> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a yield-increasing agent for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, and when component (B) is set to 1, the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000.

[0157] <35> Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for the manufacture of a yield-increasing agent for legumes contained in beans, wherein the (B) antioxidant is at least one selected from tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, and the (C) organic solvent is at least one selected from alcohols having a carbon number of 5 or less and dimethyl sulfoxide, and when component (B) is set to 1, the proportion (mass ratio) of the (C) organic solvent is 4 to 100,000.

[0158] <36> The use as described in any one of <30> to <35>, wherein, with respect to the ratio (mass ratio) of (A) ascorbic acid or a salt thereof and (C) an organic solvent, when component (B) is set to 1, component (A) is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, and component (C) is 4 to 100,000, preferably 10 to 50,000, more preferably 20 to 20,000, and further preferably 50 to 10,000.

[0159] <37> The use according to any one of <30> to <36>, wherein the (B) antioxidant further comprises one or more selected from the group consisting of sodium sulfite, potassium sulfite, glutathione and uric acid.

[0160] <38> The use according to any one of <30> to <37>, wherein the (C) organic solvent is at least one selected from isobutyl alcohol and dimethyl sulfoxide.

[0161] <39> The use according to any one of <30> to <38>, further comprising (D) a surfactant.

[0162] <40> The use according to <39>, wherein the ratio (mass ratio) of the surfactant (D) is preferably 10 to 300,000, more preferably 50 to 100,000, and even more preferably 100 to 5,000, relative to the component (B).

[0163] <41> The use according to <39> or <40>, wherein the surfactant (D) comprises at least a nonionic surfactant or an anionic surfactant.

[0164] <42> The use according to <41>, wherein the surfactant (D) further comprises an anionic surfactant in addition to the nonionic surfactant.

[0165] <43> The use as described in <42>, wherein (D) the nonionic surfactant is at least one selected from sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters and polyoxyethylene alkyl ethers, and the anionic surfactant is at least one selected from alkyl sulfate ester salts and fatty acid salts.

[0166] <44> The use according to any one of <30> to <43>, further comprising (E) a chelating agent.

[0167] <45> The use according to <44>, wherein, with respect to the ratio (mass ratio) of (A) ascorbic acid or a salt thereof, (C) an organic solvent, (D) a surfactant and (E) a chelating agent, when component (B) is taken as 1, component (A) is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, component (C) is 4 to 100,000, preferably 10 to 50,000, more preferably 20 to 20,000, further preferably 50 to 10,000, component (D) is preferably 10 to 300,000, more preferably 50 to 100,000, further preferably 100 to 5,000, and component (E) is preferably 0.01 to 100, more preferably 0.1 to 100, further preferably 1 to 10, further preferably 1 to 10.

[0168] <46> The use according to <44> or <45>, wherein the (E) chelating agent is at least one selected from ethylenediaminetetraacetic acid and ethylenediamine-N,N′-disuccinic acid.

[0169] <47> The use according to any one of <30> to <38>, comprising a first agent containing the above-mentioned component (A) and a second agent containing the above-mentioned components (B) and (C), and the two are combined when used.

[0170] <48> In <1>, <2>, <5>, <6>, <23>, <25>, <30>, <31>, <34> and <35>, the legume contained in the beans is preferably a legume selected from the group consisting of soybean, red bean, chickpea, lotus root, kidney bean, peanut, broad bean, pea, runner bean, lima bean, mung bean, cowpea, lentil and sword bean, more preferably a legume selected from the group consisting of soybean, red bean, kidney bean, pea, broad bean, chickpea and lotus root, and even more preferably soybean.

[0171] [Example]

[0172] Test Example 1 Effect of water dissolving ascorbic acid on application effect

[0173] An aqueous ascorbic acid solution in which ascorbic acid was dissolved in water and an aqueous ascorbic acid solution in which ascorbic acid was dissolved in deionized water were prepared, and the effect of the water in which the ascorbic acid was dissolved on the application effect was examined in soybean (Glycine max).

[0174] The effect of spraying 500 ppm by mass of ascorbic acid on the soybean yield was evaluated. The test plots 1 to 3 evaluated were as follows.

[0175] Partition 1: Control (no application)

[0176] 2nd zone: Spray 500 ppm by mass of ascorbic acid on the leaves (dissolved in tap water)

[0177] Partition 3: Spray 500 ppm by mass of ascorbic acid on the leaves (dissolved in deionized water)

[0178] (1) Cultivation conditions

[0179] Soybean cultivation was carried out in farmland in Tochigi Prefecture, and the soybean cultivar used was "Sato no Hohoemi". The ridge width of 2m in the farmland excluding the periphery was set as one section, and foliar spraying was carried out. For one test area, 12 sections were tested in the control area (test area 1), and 3 sections were tested in the ascorbic acid application area (test areas 2 and 3). An average of 19 individuals were cultivated in the range of 2m set as one section.

[0180] (2) Foliar spraying and determination of seed weight

[0181] As ascorbic acid, "Food Additive Grade Vitamin C (L-Ascorbic Acid) Fine Mesh Type SSS" produced by Fuso Chemical Industry Co., Ltd. was used. A 500 mass ppm aqueous solution of ascorbic acid was dissolved in tap water or deionized water to prepare it. Using a dry cell sprayer (GT-5HS, Koshin Co., Ltd.), 20 mL of the ascorbic acid aqueous solution was sprayed on each soybean plant in a manner of spraying the entire soybean plant body. Spraying was carried out on the 60th day after sowing, which is a period of growth stage equivalent to the grain swelling period. Harvesting was carried out on the 108th day after sowing. All grains were recovered from each individual after harvesting and dried at 100°C for 48 hours. The mass of dry grains was measured as yield data.

[0182] (3) Results

[0183] After calculating the average grain weight of each individual in one partition, calculate the average and standard deviation of the three partitions ( Figure 1 ). The graph in the figure represents the mean ± standard deviation. It shows that when the same concentration of ascorbic acid is sprayed on the leaves, the effect on seed weight is different when tap water and deionized water are used. In other words, it is clear that the technical problem that the effect of ascorbic acid disappears when using water (tap water) used in agricultural sites is clarified.

[0184] Test Example 2 Solubility of Butylated Hydroxytoluene in Various Solvents

[0185] The amount of BHT dissolved in 100 μL of isobutyl alcohol was changed. It was confirmed that the BHT was uniformly dissolved until the BHT reached 25 mg, that is, until the ratio of BHT:isobutyl alcohol = 1:4 ( Figure 2 ). The solubility of the hydroxyl group was also confirmed in other solvents up to a ratio of BHT:solvent = 1:4. Among the solvents studied, the hydroxyl group was uniformly soluble in ethanol, methanol, 2-propanol, 1-butanol, and dimethyl sulfoxide (DMSO) except glycerol ( Figure 3 ). It is known that BHT is poorly soluble in water. Here, each reagent used was a product of Fujifilm Wako Pure Chemical Industries, Ltd.

[0186] Example 1 Preparation of a spray liquid having plant growth promoting, nodule activity promoting and yield increasing effects

[0187] (1) Preparation of dual-dose composition

[0188] The two-dose composition shown in Table 1 was prepared using the following ingredients. One or more selected from BHT, BHA and tocopherol as component (B) was preliminarily dissolved in component (C), and then component (D) was mixed to prepare the second dose. Next, one or more selected from component (A) and component (E) were mixed to prepare the first dose. Each composition example shown in Table 2 was prepared in the form of a two-dose composition in which component (A), component (B), component (C), component (D) and component (E) were mixed in the amounts shown in the table.

[0189] (2) Preparation of spray solution and solubility evaluation

[0190] The prepared two-component composition was dissolved in 100 mL of tap water heated to 30° C., and the solubility of the spray solution was evaluated. That is, after the second component was dissolved in heated tap water, the first component was dissolved and the solubility was evaluated.

[0191] (3) Reagents used

[0192] Ascorbic acid, "Food Additive Grade Vitamin C (L-Ascorbic Acid) Fine Mesh Type SSS" manufactured by Fuso Chemical Industries, Ltd. was used. Sodium ascorbate, tocopherol, BHT, BHA, ethanol, methanol, 2-propanol, isobutanol, 1-butanol, DMSO, and citric acid were products of Fujifilm Wako Pure Chemical Industries, Ltd. EDTA was a product of Dojin Chemical Laboratories, Ltd. As surfactants, sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, polyoxyethylene lauryl ether, polyoxyethylene alkyl ether, and polyethylene glycol monolaurate were used.

[0193] (4) Results of solubility evaluation

[0194] The solubility of the spray liquid prepared using each composition in aqueous solution was evaluated for appearance according to the following criteria.

[0195] ○: No residual solution, a uniform aqueous solution.

[0196] △: Some residual solution was left, but the mixture became uniform immediately after stirring.

[0197] ×: Dissolved residue is visible and the mixture is not uniform immediately after stirring.

[0198] The results in Table 2 show that in Composition Examples 1 to 17, a uniform spray liquid suitable for spraying as a plant growth promoter can be obtained.

[0199] [Table 1]

[0200]

[0201]

[0202] Example 2 Evaluation of nodule weight and nodule activity

[0203] (1) Soil preparation and sowing

[0204] The medium-term fertilizer-effective culture soil (TAKII water-containing cell culture soil medium-term fertilizer-effective type, TAKII & CO., LTD.) and vermiculite particles (Akagi-Engei Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a plastic seedling pot (diameter 10.5 cm, height 9 cm). Soybean seeds used "Enrei" (purchased from Nikko Seed Co., Ltd.). After supplying 250 mL of tap water to each pot, 2 seeds were sown per pot, one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of replicates in each test area was 6 (n = 6).

[0205] (2) Inoculation of Rhizobia

[0206] A solid medium was prepared by adding 1.5% agar (Wako Pure Chemical Industries, Ltd.) to a yeast-mannitol (YM) medium (K2HPO4 0.5 g, MgSO4·7H2O 0.2 g, NaCl 0.1 g, yeast extract 0.4 g, mannitol 10 g, distilled water 1 L (pH 6.8)), and the soybean rhizobium (Bradyrhizobium japonicum) NBRC14783T strain was grown on the solid medium. A platinum loop of the grown rhizobia was taken and inoculated into 50 mL of YM liquid medium in a 500 mL Sakaguchi flask, and cultured at 30°C with shaking for 36 hours. A rhizobium culture solution with a turbidity OD600 value of the bacterial cells of about 0.3 was prepared. After sowing, 1 mL of the rhizobium culture solution was dripped onto the seeds using a micropipette.

[0207] (3) Cultivation conditions

[0208] The cultivation from sowing to thinning was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 30°C in light period / 25°C in dark period, and the humidity was set to 50%. 7 days after sowing, thinning was performed so that there was one plant per pot. After thinning, cultivation was carried out outdoors. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray again to immerse the lower 5 cm of the pot.

[0209] (4) Foliar spraying

[0210] The spray liquid shown in Table 3 was prepared, and 6.7 mL was sprayed per plant using a spray pot on the 14th day after sowing. Foliar spraying was performed only once, and the 7 test plots were studied.

[0211] Regarding the preparation of the spray liquid, BHT in component (B) was pre-dissolved in component (C) DMSO, and then component (C) and component (D) were mixed to prepare the second dose. Next, component (A), uric acid (antioxidant) and component (E) were mixed to prepare the first dose. After the second dose was dissolved in water, the first dose was dissolved to prepare the spray liquid.

[0212] Here, ascorbic acid used is "Food Additive Grade Vitamin C (L-Ascorbic Acid) Fine Mesh Type SSS" produced by Fuso Chemical Industries, Ltd. BHT, uric acid, DMSO and EDTA used are products of Fujifilm Wako Pure Chemical Industries, Ltd., and EDDS used is "Chelest EDDS-35" produced by Chelest Co., Ltd. As component (C), DMSO and isobutyl alcohol are used. In component (D), sorbitan monolaurate or sorbitan fatty acid ester, polyoxyethylene resin acid ester are used as nonionic surfactants. In addition, in component (D), fatty acid soap potassium salt is used as an anionic surfactant.

[0213]

[0214] (5) Method for recovering exudate for measuring urea nitrogen as an indicator of nodule activity

[0215] After 21 days of cultivation, the soybean stems were cut at the cotyledonary node using pruning shears, covered with cotton balls #10 (Kawamoto Sangyo Co., Ltd.) filled in 1.5 mL microtubes (Eppendorf Co., Ltd.), and the exudate flowing out from the cut surface of the stems was collected for 2 hours. The cotton balls after the exudate was collected were stored in a refrigerator at -80°C. In addition, the amount of exudate was calculated by comparing the weight of the cotton balls before and after the exudate was collected.

[0216] (6) Extraction of exudate components recovered from cotton balls

[0217] The exudate components contained in the cotton ball were eluted with ultrapure water and filtered using a Micro Biospin column (Bio-Rad Laboratories, Inc.). The residual filtrate in the column was centrifuged and recovered at 15,000 rpm for 1 minute using a centrifuge (CR15RN, Hitachi, Ltd.). The filtrate diluted 1,000 times was quantified.

[0218] (7) Apparatus and measurement method used for quantification of urea nitrogen content

[0219] The HPLC apparatus and mass spectrometer used were Agilent 1260 Infinity LC system (Agilent Technologies, Inc.) and AB SCIEX TripleQuad 4500 system (AB Sciex Pte. Ltd.), respectively. The column used was Scherzo SS-C18 (100 mm×2 mm, 3 μm) (IMTAKT Co., Ltd.), and the oven temperature was set to 40°C. 5 μL of the appropriately diluted sample was injected, the flow rate was 0.5 mL / min, and the eluents used were 0.1% formic acid aqueous solution (eluent A) and 50 mM ammonium acetate / methanol solution (eluent B). After equilibration with eluent A: eluent B = 95:5, the sample was supplied, and after 5 minutes, urea nitrogen was eluted by a linear gradient reaching eluent A: eluent B = 80:20.

[0220] (8) Standards used and quantification

[0221] The amount of urea nitrogen in the exudate was analyzed by LC-MS. Allantoin (Tokyo Chemical Industry Co., Ltd.) and allantoic acid (Toronto Research Chemicals Inc.) were used as standards. The standards were analyzed by LC-MS, and a calibration curve was prepared in the range of 10-1000 ppb.

[0222] For each sample, allantoin and allantoic acid were identified based on the consistency of retention time, accurate mass, and MS / MS spectrum with each reagent. In addition, allantoin and allantoic acid in the sample were quantified based on the calibration curve to determine the total amount per plant.

[0223] (9) Results

[0224] The results of nodule weight and nodule activity are shown in Figure 4 and Figure 5 The highest and lowest values ​​were removed from the six replicates in each test area, and the mean and standard deviation were calculated using the data from four replicates. The graph in the figure represents the mean ± standard deviation.

[0225] In Test Example 1, the effect of water quality differences in dissolving ascorbic acid on soybean yield was shown. According to Test Areas 1 to 3 of this example, even in the early development stage of soybeans, it was confirmed that the nodule weight and nodule activity increased when ascorbic acid was dissolved in deionized water, and the application effect of ascorbic acid decreased when it was dissolved in tap water. According to the comparison between Test Areas 4 and 5, it was shown that in order to restore the nodule activity reduced by dissolving ascorbic acid in tap water, it was necessary to add an organic solvent. In addition, according to the comparison between Test Areas 6 and 7, it was taught that EDTA among chelating agents was more conducive to the recovery of nodule activity.

[0226] Example 3 Study on the Mixing Ratio of Ascorbic Acid and Antioxidants

[0227] The preparation and sowing of soil, inoculation of rhizobia, the method for recovering the exudate for measuring the amount of urea nitrogen as an indicator of nodule activity, the experimental operations before quantifying the exudate, the apparatus and measurement method used for quantifying the amount of urea nitrogen, the standards used and the quantitative items were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0228] (1) Cultivation conditions

[0229] The cultivation was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 30°C in light period / 25°C in dark period, and the humidity was set to 50%. After 7 days from sowing, thinning was performed so that there was one plant per pot. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to immerse the lower 5 cm of the pot.

[0230] (2) Foliar spraying

[0231] The spray solution shown in Table 4 below was prepared in the same manner as in Example 2, and 6.7 mL was sprayed per plant using a spraying pot on the 14th day after sowing. Foliar spraying was performed only once, and seven test plots were studied (Table 4). The concentration of ascorbic acid in the spray solution was adjusted by fixing the amount of water sprayed and changing the amount of ascorbic acid per plant. The types of reagents used were the same as in Example 2.

[0232]

[0233] (3) Results

[0234] The results of the nodulation activity assay are shown in Figure 6 and Figure 7 Graphs in the figures represent mean ± standard deviation.

[0235] exist Figure 6 and Figure 7 In the experiment, when ascorbic acid dissolved in tap water was sprayed on the leaves, the nodule activity did not change or even decreased compared to when no application was done. In the test area 3 with a composition not containing ascorbic acid, the nodule activity decreased, but by adding ascorbic acid to the composition of the test area 3, the nodule activity was restored or improved. In the experiment on the amount of water to be sprayed using a sprayer (such as a plunger pump sprayer), the nodule activity in the test area 4 with ascorbic acid:BHT=500:1 was the highest, and the nodule activity tended to decrease in the test area 7 with ascorbic acid:BHT=20,000:1. In addition, when ascorbic acid:BHT=20,000:1, Figure 8 As shown, the leaf color loss problem occurred, indicating that the mixing ratio of ascorbic acid, component (B), and component (C) is preferably 100 to 20,000:1:2 to 100. In addition, when isobutyl alcohol was used as component (C), it was also shown to have a nodule activity recovery effect.

[0236] Example 4: Effect of increasing soybean yield in farmland

[0237] The effect of foliar spraying of a composition containing 1,500 ppm ascorbic acid on soybean yield was evaluated. The foliar spray was applied only once and eight test plots were studied (Table 5).

[0238]

[0239] (1) Cultivation conditions

[0240] Soybean cultivation was carried out in farmland in Tochigi Prefecture, and the soybean cultivar used was "Sato no Hohoemi". The ridge width of 2m excluding the periphery of the farmland was set as one section, and foliar spraying was carried out. Three sections were tested for one test area. An average of 10 individuals were cultivated in a range of 2m set as one section.

[0241] (2) Foliar spraying and determination of seed weight

[0242] Polyoxyethylene lauryl ether was used as a nonionic surfactant in addition to the reagents described in Example 2. Urea, potassium dihydrogen phosphate, sodium sulfite, glutathione, and EDTA were products of Fujifilm Wako Pure Chemical Industries, Ltd.

[0243] The reagent was dissolved in tap water or deionized water to prepare a spray solution. Using a dry cell sprayer (GT-5HS, Koshin Co., Ltd.), 6.7 mL of ascorbic acid aqueous solution was sprayed on each soybean plant in the form of a spray on the entire soybean plant. Spraying was carried out on the 75th day after sowing, and the growth stage was a period equivalent to the grain hypertrophy period. Harvesting was carried out on the 173rd day after sowing. All grains were recovered from each individual after harvesting and dried at 100°C for 48 hours. The mass of dry grains was measured as yield data.

[0244] (3) Results

[0245] After calculating the average grain weight of each individual in one partition, calculate the average and standard deviation of the eight partitions ( Fig. 9 ). Graphs in the figures represent mean ± standard deviation.

[0246] It was confirmed again that when the same concentration of ascorbic acid was sprayed on the leaves, the effect on seed weight was different when tap water and deionized water were used. It was shown that by adding component (B), component (C) and component (D) to ascorbic acid, a yield-increasing effect can be obtained even when tap water is used. It was also shown that when polyoxyethylene lauryl ether was used as component (D), a yield-increasing effect was also obtained.

[0247] Example 5 Evaluation of nodulation activity by administration of high concentrations of ascorbic acid

[0248] The preparation and sowing of soil, inoculation of rhizobia, cultivation conditions, the method for recovering the exudate for measuring the amount of urea nitrogen as an indicator of nodule activity, the experimental operations before quantifying the exudate, the apparatus and determination method used for quantifying the amount of urea nitrogen, the standards used and the quantitative items were the same as those in Example 3, and soybean cultivation was carried out for 21 days.

[0249] (1) Foliar spraying

[0250] Foliar spray treatment was carried out on the 14th day after sowing. In addition to the reagents described in Examples 2, 3 and 4, polyethylene glycol monolaurate and glycerol fatty acid esters were used as non-ionic surfactants. The spray solution was prepared by dissolving the reagents in tap water or deionized water. Foliar spraying was carried out only once, and 8 test areas containing high concentrations of ascorbic acid were studied assuming aerial spraying (Table 6). Test areas 2 and 3 were sprayed with 6.7 mL per plant using a spray bottle, and test areas 4 to 8 were sprayed with 0.125 mL per plant.

[0251]

[0252] (2) Results

[0253] The results of the nodulation activity assay are shown in Fig.10The highest and lowest values ​​were removed from the six replicates in each test area, and the average and standard deviation were calculated using the data from four replicates. The graph in the figure represents the average ± standard deviation.

[0254] exist Fig.10 In the test plots 4 and 5 where ascorbic acid was sprayed with a small amount of water, there was a tendency for soybean nodule activity to increase compared with test plots 2 and 3. Regardless of the amount of water used for spraying, the nodule activity was improved when deionized water was used to dissolve ascorbic acid compared with tap water. According to the comparison between test plots 5 and 6, the nodule activity was highest by adding BHT as compound (B), DMSO as compound (C), and polyethylene glycol monolaurate as compound (D) to ascorbic acid. On the other hand, when the ascorbic acid concentration was increased to 300,000 ppm in test plot 7, the nodule activity decreased compared with test plot 6, indicating that the ascorbic acid concentration is preferably 300,000 ppm or less when spraying with a small amount of water. In addition, according to the comparison between test plots 8 and 6, it was shown that compound (C) was better using DMSO than isobutanol. It was also shown that when polyethylene glycol monolaurate was used as compound (D), it also had a root nodule activity recovery effect.

[0255] Example 6 Evaluation of initial growth indicators of red beans

[0256] (1) Soil preparation and sowing

[0257] The medium-term fertilizer-effective culture soil (TAKII water-containing cell culture soil medium-term fertilizer-effective type, TAKII & CO., LTD.) and vermiculite particles (Akagi-Engei Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a plastic seedling pot (diameter 10.5 cm, height 9 cm). Red bean seeds used "Tamba Dainagon Azuki" (purchased from Nikko Seed Co., Ltd.). After supplying 250 mL of tap water to each pot, 2 seeds were sown per pot, and sown one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of replicates in each test area was 6 (n = 6).

[0258] (2) Inoculation of Rhizobia

[0259] A solid medium was prepared by adding 1.5% agar (Wako Pure Chemical Industries, Ltd.) to a yeast-mannitol (YM) medium (K2HPO4 0.5 g, MgSO4·7H2O 0.2 g, NaCl 0.1 g, yeast extract 0.4 g, mannitol 10 g, distilled water 1 L (pH 6.8)), and the soybean rhizobium (Bradyrhizobium japonicum) NBRC14783T strain was grown on the solid medium. A platinum loop of the grown rhizobia was taken and inoculated into 50 mL of YM liquid medium in a 500 mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of the bacterial cells was prepared to be about 0.3. After sowing, 1 mL of the rhizobium culture solution was dripped onto the seeds using a micropipette.

[0260] (3) Cultivation conditions

[0261] The cultivation was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 25°C in light period / 20°C in dark period, and the humidity was set to 50%. After 7 days from sowing, thinning was performed so that there was one plant per pot. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to immerse the lower 5 cm of the pot.

[0262] (4) Foliar spraying

[0263] Prepare a spray solution, and spray 6.7 mL per plant once with a spray bottle on the 21st day after sowing. For the preparation of the spray solution, BHT was dissolved in isobutyl alcohol, the isobutyl alcohol was mixed with sorbitan monolaurate, and then dissolved in water. Then, ascorbic acid was dissolved in water to prepare the solution. The reagents used were the same as those in Example 2. The test areas 1 to 4 evaluated were as follows.

[0264] On the 28th day after sowing, the plant bodies were dried at 90°C for 24 hours, and then the above-ground dry weight and the underground dry weight of the plant bodies were measured.

[0265] Partition 1: Control (no application)

[0266] 2nd zone: Spray 1,500 ppm by mass of ascorbic acid on the leaves (dissolved in tap water)

[0267] Zone 3: Spray 1,500 ppm by mass of ascorbic acid on the leaves (dissolved in deionized water)

[0268] 4th zone: Foliar spraying of ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + isobutyl alcohol 50 mass ppm + sorbitan monolaurate 350 mass ppm (with tap water) (5) Results

[0269] The above-ground dry weight and underground dry weight are as follows: Fig.11 and Fig.12 The graphs in the figures represent the mean ± standard deviation.

[0270] According to Test Example 1 and Example 2, when the same concentration of ascorbic acid is sprayed on the soybean leaf surface, the effects on seed weight and initial growth index are different when tap water and deionized water are used. In red beans, as with soybeans, it is shown that the effects on initial growth indexes are different depending on the quality of the water in which ascorbic acid is dissolved. According to the comparison between Test Areas 2 and 4, by adding component (B), component (C) and component (D) to ascorbic acid, a growth promoting effect can be obtained even when tap water is used.

[0271] Example 7 Evaluation of initial growth indicators of chickpea

[0272] (1) Soil preparation and sowing

[0273] The medium-term fertilizer-effective culture soil (TAKII water-containing cell culture soil medium-term fertilizer-effective type, TAKII & CO., LTD.) and vermiculite particles (Akagi-Engei Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a plastic seedling pot (diameter 10.5 cm, height 9 cm). Chickpeas used seeds of the kabuli species (purchased from Nikko Seedling Co., Ltd.). After supplying 250 mL of tap water to each pot, 2 seeds were sown per pot, and sown one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of replicates in each test area was 6 (n = 6).

[0274] (2) Inoculation of Rhizobia

[0275] A solid medium was prepared by adding 1.5% agar (Wako Pure Chemical Industries, Ltd.) to a yeast-mannitol (YM) medium (K2HPO4 0.5 g, MgSO4·7H2O 0.2 g, NaCl 0.1 g, yeast extract 0.4 g, mannitol 10 g, distilled water 1 L (pH 6.8)), and the chickpea rhizobium (Mesorhizobium ciceri) NBRC100389T strain was grown on the solid medium. A platinum loop of the grown rhizobia was taken and inoculated into 50 mL of a YM liquid medium in a 500 mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of the bacterial cells was prepared to be about 0.3. After sowing, 1 mL of the rhizobium culture solution was dripped onto the seeds using a micropipette.

[0276] (3) Foliar spraying

[0277] The cultivation conditions and the method of foliar spraying were the same as those in Example 6 (the example of adzuki beans). The evaluation test plots 1 to 4 were as follows.

[0278] On the 28th day after sowing, the plant bodies were dried at 90°C for 24 hours, and the dry weight of the aboveground part of the plant bodies was measured.

[0279] Partition 1: Control (no application)

[0280] 2nd zone: Spray ascorbic acid 1,500 ppm by mass (dissolved in tap water) on the leaves

[0281] Zone 3: Spray 1,500 ppm by mass of ascorbic acid on the leaves (dissolved in deionized water)

[0282] 4th zone: foliar spray of ascorbic acid 1,500 ppm by mass + BHT 1 ppm by mass + isobutyl alcohol 50 ppm by mass + sorbitan monolaurate 350 ppm by mass (dissolved in tap water)

[0283] (4) Results

[0284] The results of the aboveground dry weight determination are shown in Fig.13 The highest and lowest values ​​were removed from the six replicates in each test area, and the mean and standard deviation were calculated using the data from four replicates. The graph in the figure represents the mean ± standard deviation.

[0285] According to Test Example 1 and Example 2, when the same concentration of ascorbic acid is sprayed on the soybean leaf surface, the effects on seed weight and initial growth indexes are different when tap water and deionized water are used. In chickpea, as in soybean, it is shown that the effect on initial growth indexes is different depending on the quality of water in which ascorbic acid is dissolved. According to the comparison between Test Areas 2 and 4, by adding component (B), component (C) and component (D) to ascorbic acid, a growth promoting effect can be obtained even when tap water is used.

[0286] Example 8: Evaluation of the seed weight of Lotus japonicus

[0287] (1) Soil preparation

[0288] Mid-term fertilizer-effective culture soil (TAKII Aqueous Cell Culture Soil Mid-term Fertilizer-effective Type, TAKII & CO., LTD.) and vermiculite fine particles (Akagi-Engei Co., Ltd.) were mixed at a volume ratio of 1:1, and the soil was filled into a plastic seedling pot (6 cm in diameter, 5.5 cm in height).

[0289] (2) Germination and sowing

[0290] The seeds of Lotus japonicus were sown using the Miyakojima MG-20 system. 1 mL of concentrated sulfuric acid (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added to a 2 mL microtube (Eppendorf Co., Ltd.) with about 1 / 3 of the seeds added, and the tube was left to stand for 10 minutes. After washing with tap water 5 times, the tube was immersed in water for 2 hours. After supplying 100 mL of tap water to each pot, 3 seeds that had been treated for germination were sown one by one in each pot at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions in each test area was 8 (n=8).

[0291] (3) Inoculation of Rhizobia

[0292] A solid medium was prepared by adding 1.5% agar (Wako Pure Chemical Industries, Ltd.) to a yeast-mannitol (YM) medium (K2HPO4 0.5 g, MgSO4·7H2O 0.2 g, NaCl 0.1 g, yeast extract 0.4 g, mannitol 10 g, distilled water 1 L (pH 6.8)), and Mesorhizobium loti MAFF303099 (ML GUS) strain was grown on the solid medium. A platinum loop of the grown rhizobia was taken and inoculated into 50 mL of YM liquid medium in a 500 mL Sakaguchi flask, and cultured at 30°C with shaking for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of the bacterial cells was prepared to be about 0.3. After sowing, 1 mL of the rhizobium culture solution was dripped onto the seeds using a micropipette.

[0293] (4) Cultivation conditions

[0294] The cultivation was carried out indoors, and the cultivation conditions were set as a 16-hour photoperiod, 25°C, LED light source (OHGETS Co., Ltd., model: VGL-1200W), and light intensity set to 400-440 μmol / m 2 / s. After 10 days from sowing, thinning was performed so that there was one plant per pot. For watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to submerge the lower 5 cm of the pot. After 3 weeks from sowing, a 1,000-fold dilution of a high-grade flowering promoter (Hyponex Japan Corp., Ltd.) was added to the irrigation water once a week.

[0295] (5) Foliar spraying

[0296] The spray solution shown in Table 7 was prepared, and 6 weeks after sowing, 6.7 mL was sprayed per plant in test plots 2 to 6 only once, and 0.125 mL was sprayed per plant in test plots 7 to 9 only once. The spray solution shown in Table 13 was prepared, and 6.7 mL was sprayed per plant in a spray can on the 14th day after sowing. Foliar spraying was performed only once, and 7 test plots were studied.

[0297] Regarding the preparation of the spray liquid, component (B) BHT was preliminarily dissolved in component (C), and then component (C) and component (D) were mixed to prepare the second dose. Next, component (A) and other components were mixed to prepare the first dose. After dissolving the second dose in the water to be used, the first dose was dissolved to prepare the spray liquid. The mature grains were harvested in sequence, and the cultivation was terminated 16 weeks after sowing. The number of pods and seed weight of each plant were measured.

[0298]

[0299] (6) Results

[0300] The results of the pod number determination are shown in Fig.14 The results of seed weight determination are shown in Fig.15 Graphs in the figures represent mean ± standard deviation.

[0301] According to Test Example 1 and Example 2, when the same concentration of ascorbic acid is sprayed on the soybean leaves, the effects on seed weight are different when tap water and deionized water are used. In Lotus japonicus, as in soybean, it is shown that the effect on seed weight is different depending on the quality of the water in which ascorbic acid is dissolved. According to the comparison between Test Areas 3 and 4, by adding components (B), (C) and (D) to ascorbic acid, the number of pods and seed weight are increased even when tap water is used. According to Test Area 5, when fertilizer components are added to the composition, an increase in yield can also be obtained. In addition, as shown in Test Area 6, a sufficient application effect is obtained when the concentration of the composition is halved.

[0302] According to Example 5, when a high-concentration composition is applied to soybean, the nodule activity during the initial growth is improved. In Lotus japonicus, as in soybean, by adding component (B), component (C) and component (D) to ascorbic acid, a yield-increasing effect can be obtained even when a high-concentration spray is performed with a small amount of tap water.

[0303] Example 9 Evaluation of the weight of broad beans

[0304] (1) Soil preparation and cultivation conditions

[0305] After filling 2L of flower pot bottom stone in a flower pot (65.3cm wide, 24.5cm deep, 18.5cm high), fill it with 9L of medium-term fertilizer-effective culture soil (TAKII water-containing cell culture soil medium-term fertilizer-effective type, TAKII & CO., LTD.). After supplying 2L of tap water to each flower pot, transplant 3 broad bean seedlings purchased at a home center (variety: Rende Ichicun, TAKII & CO., LTD.). In addition, the number of replicates in each test area is 3 (n=3). Cultivation is carried out outdoors.

[0306] (2) Foliar spraying

[0307] The spray solution was prepared and 6.7 mL was sprayed per plant using a spray bottle on the 51st day after the seedlings were planted. Foliar spraying was performed only once, and four test plots were studied.

[0308] Regarding the preparation of the spray liquid, the BHT in the component (B) was preliminarily dissolved in the component (C), and then the component (C) and the component (D) were mixed to prepare the second agent. Then, the component (A) and the other components were mixed to prepare the first agent. After the second agent was dissolved in the water to be used, the first agent was dissolved to prepare the spray liquid. The reagent type was the same as that in Example 1. The test areas 1 to 4 evaluated were as follows.

[0309] The mature seeds were harvested in sequence, and the cultivation was terminated 96 days after the start of planting. The number of beans and the weight of seeds of each plant were measured.

[0310] Partition 1: Control (no application)

[0311] Partition 2: Spray 1,500 ppm by mass of ascorbic acid on the leaves (dissolved in deionized water)

[0312] 3rd zone: Spray 1,500 ppm by mass of ascorbic acid on the leaves (dissolved in tap water)

[0313] 4th zone: foliar spray of ascorbic acid 1,500 ppm by mass + BHT 1 ppm by mass + isobutyl alcohol 50 ppm by mass + sorbitan monolaurate 350 ppm by mass (dissolved in tap water)

[0314] (3) Results

[0315] The results of the determination of the number of beans per individual are shown in Fig.16 The results of seed weight determination are shown in Fig.17 . The graph in the figure represents the mean ± standard deviation. Comparison between test plots 1 and 4 showed that by adding component (B), component (C) and component (D) to ascorbic acid, the number of beans per plant increased by 7.5%, the grain weight increased by 32.9%, and the yield of broad beans was also increased. In addition, comparison between test plots 3 and 4 showed that by adding component (B), component (C) and component (D) to ascorbic acid, the yield-increasing effect was improved.

[0316] Example 10 Storage stability test of a composition having plant growth promoting, nodule activity promoting and yield increasing effects

[0317] (1) Preparation of single-dose or double-dose compositions

[0318] As component (A), 20 g of ascorbic acid, 13.3 mg of BHT, 0.67 mL of isobutyl alcohol, and 4.7 mL of sorbitan monolaurate are used as component (D), and a single-dose or double-dose composition is prepared according to the following method. Component (B) is dissolved in component (C), and then component (D) is mixed, and component (A) is mixed with the resulting mixture to prepare a single-dose composition. The preparation of the double-dose composition is carried out according to the following method. Component (B) is pre-dissolved in component (C), and then component (D) is mixed to prepare a second dose. Next, only component (A) is used as the first dose. The types of reagents are the same as those in Example 1.

[0319] (2) Storage stability test

[0320] Half of the prepared single-dose or double-dose composition was taken and injected into two transparent glass bottles. The bottles were stored in a refrigerator set at 4°C or a storage box set at 50°C for 1 week. After 1 week, the appearance was evaluated, and the aqueous solution solubility of the stored preparations when diluted 500 times in water was evaluated.

[0321] (3) Results of the evaluation of the appearance and solubility of the preparation

[0322] The results of the appearance evaluation of the preparations are shown in Fig.18 The left side of the photo shows the first and second doses of the two-dose composition after storage at 50°C, the middle of the photo shows the first and second doses of the two-dose composition after storage at 4°C, and the right side of the photo shows the single-dose composition after storage at 50°C and the single-dose composition after storage at 4°C.

[0323] according to Fig.18 The results showed that there was no difference in appearance between the samples of the two-dose composition stored at 4°C and the samples stored at 50°C. However, the appearance color of the single-dose composition was different due to the storage temperature. It also showed that a uniform spray liquid suitable for spraying as a plant growth promoter can be obtained in any of the single-dose and double-dose formulations. The research results taught that the single-dose formulation has a large change in color change during storage, and the double-dose formulation has an advantage from the perspective of appearance.

[0324] Example 11 Study on the Mixing Ratio of Organic Solvents

[0325] The preparation and sowing of soil, inoculation of rhizobia, the method for recovering the exudate for measuring the amount of urea nitrogen as an indicator of nodule activity, the experimental operations before quantifying the exudate, the apparatus and measurement method used for quantifying the amount of urea nitrogen, the standards used and the quantitative items were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0326] (1) Cultivation conditions

[0327] The cultivation was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 30°C in light period / 25°C in dark period, and the humidity was set to 50%. After 7 days from sowing, thinning was performed so that there was one plant per pot. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to immerse the lower 5 cm of the pot.

[0328] (2) Foliar spraying

[0329] The spray solution shown in Table 8 was prepared. On the 14th day after sowing, 6.7 mL was sprayed once per plant in test plots 1 to 5 and 0.125 mL was sprayed once per plant in test plots 6 to 8 using a spraying pot. Foliar spraying was performed only once, and 7 test plots were studied (Table 8). The types of reagents used were the same as in Example 5.

[0330]

[0331] (3) Results

[0332] The results of the nodulation activity assay are shown in Fig.19 Graphs in the figures represent mean ± standard deviation.

[0333] Comparison between test area 2 and test areas 3 to 8 showed that by adding component (B), component (C) and component (D) to ascorbic acid, a higher effect of improving root nodule activity can be obtained compared with spraying ascorbic acid dissolved in tap water alone. It was also shown that the effect can be exerted even when the mixing ratio of component (B) to component (C) is set to 1:10 to 10000.

[0334] Example 12 Evaluation of initial growth indicators at low concentrations of antioxidants

[0335] The preparation of soil, sowing, and inoculation of rhizobia were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0336] (1) Cultivation conditions

[0337] The cultivation was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 30°C in light period / 25°C in dark period, and the humidity was set to 50%. After 7 days from sowing, thinning was performed so that there was one plant per pot. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to immerse the lower 5 cm of the pot.

[0338] (2) Foliar spraying

[0339] Prepare the spray solution, and spray 6.7 mL per plant only once on the 14th day after sowing with a spray bottle. For the preparation of the spray solution, after dissolving BHT in isobutyl alcohol, isobutyl alcohol and sorbitan monolaurate are mixed and then dissolved in water. Then, ascorbic acid is dissolved in water for preparation. The reagents used are the same as those in Example 2. The evaluated test areas 1 to 3 are as follows. On the 21st day after sowing, the plant body was dried at 90°C for 24 hours, and the dry weight of the aboveground part of the plant body was measured.

[0340] Partition 1: Control (no application)

[0341] 2nd zone: Spray ascorbic acid 1,500 ppm by mass (dissolved in tap water) on the leaves

[0342] Part 3: Spray 1,500 ppm by mass of ascorbic acid + 0.01 ppm by mass of BHT + 50 ppm by mass of isobutyl alcohol + 350 ppm by mass of sorbitan monolaurate on the leaves (dissolved in tap water)

[0343] (3) Results

[0344] The results of the aboveground dry weight determination are shown in Fig. 20 Graphs in the figures represent mean ± standard deviation.

[0345] Comparison between test plots 2 and 3 showed that even when the concentration of BHT of component (B) was set to 0.01 ppm by mass, by adding components (B), (C) and (D) to ascorbic acid, a higher growth promoting effect than that obtained by spraying ascorbic acid dissolved in tap water alone could be obtained.

[0346] Example 13 Evaluation of initial growth index when using anionic surfactant

[0347] The preparation of soil, sowing, and inoculation of rhizobia were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0348] (1) Cultivation conditions

[0349] The cultivation was carried out in an artificial weather device (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the photoperiod (light intensity 440-480 μmol / m 2 / s) 16 hours in light period / 8 hours in dark period, the temperature was set to 30°C in light period / 25°C in dark period, and the humidity was set to 50%. After 7 days from sowing, thinning was performed so that there was one plant per pot. Regarding watering, after the water in the tray placed under the pot disappeared, tap water was added to the tray to immerse the lower 5 cm of the pot.

[0350] (2) Foliar spraying

[0351] Foliar spraying was performed on the 14th day after sowing. In addition to the reagents described in Example 2, sodium lauryl sulfate, ammonium lauryl sulfate, and triethanolamine lauryl sulfate were used as anionic surfactants. The reagents were dissolved in tap water to prepare a spray solution. Foliar spraying was performed only once, and 5 test areas were studied (Table 9). Prepare the spray solution and spray 6.7 mL on each plant with a spray bottle. On the 21st day after sowing, the plant body was dried at 90°C for 24 hours, and the dry weight of the underground part of the plant body was measured.

[0352]

[0353] (3) Results

[0354] The results of the dry weight determination of the underground part are shown in Fig.21 The graph in the figure represents the mean value ± standard deviation. The comparison between test areas 2 and 3, 4, and 5 shows that even when an anionic surfactant such as sodium lauryl sulfate, ammonium lauryl sulfate, or triethanolamine lauryl sulfate is used as component (D), by adding components (B), (C), and (D) to ascorbic acid, a higher growth promoting effect can be obtained than when ascorbic acid dissolved in tap water is sprayed alone.

[0355] Example 14 Storage stability of single-dose preparation

[0356] (1) Preparation of single-dose composition

[0357] As component (A), 300 g of ascorbic acid, 0.2 g of BHT, 10 mL of isobutyl alcohol, and 70 mL of sorbitan monolaurate were used as component (D) to prepare a single-dose composition according to the following method. Component (B) was dissolved in component (C), and then component (D) was mixed, and the resulting mixture was mixed with component (A) to prepare a single-dose composition. The types of reagents were the same as those in Example 1.

[0358] (2) Preparation of powdered composition

[0359] The following carriers were used as component (F) and mixed with the single-dose composition to prepare a powder composition. The contents of the components in the obtained powder composition are shown in Table 10.

[0360] Zeolite (ZEOLITE (POWDER) produced by ZEOBUILDER CO., LTD), silica (Tokusil NP produced by Oriental Silicas Corporation), bentonite (Odosorb K-400 produced by Kurosaki Shirotoku Co., Ltd.), and thenardite (a sample with an average particle size of 25 μm obtained by dry-grinding anhydrous thenardite produced by China National Salt Huaian Salt Chemical Group Co., Ltd.).

[0361] (3) Storage stability test

[0362] After each powdered composition was stored at room temperature for 1 day, two sieves with mesh sizes of 2.0 mm and 9.5 mm and a tray were used for the composition. The sieves were stacked in order from the smaller mesh size on the tray. 30 g of the composition was added from the top 9.5 mm sieve, and the mixture was placed in a small sieve shaker (manufactured by AS ONE CORPORATION., shaking speed 3) and shaken for 10 minutes. The mass of the particles remaining on each sieve and tray was measured. The mass passing through the 2.0 mm sieve was divided by the total mass to obtain the sieving rate.

[0363] Screening rate = (mass passing through the screen / total mass) × 100

[0364] The results are shown in Table 10. As can be seen from the results in Table 10, the sieving rate of each powdery composition was higher than that of the comparative example, that is, the increase in particle size due to surface adhesion during storage was suppressed, and thus the storage stability was excellent.

[0365] (4) Anti-browning test

[0366] 5 g of each powdered composition was dispensed into a transparent glass bottle. The mixture was stored in a storage box set at 50°C for 2 weeks. After 2 weeks, the browning degree of the appearance was evaluated on a 4-level scale (0: no browning, 1: slightly visible browning, 2: visible browning, 3: obvious browning)

[0367] The results are shown in Table 10. According to the results in Table 10, in the composition to which silica was added, browning after storage was suppressed. On the other hand, browning of the appearance was observed in the other compositions. The results of the study showed that adding silica was superior from the viewpoint of appearance stability.

[0368]

Claims

1. A growth promoter for leguminous plants contained in beans, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

2. A growth promoter for leguminous plants contained in beans, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

3. A nodule activity promoter, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

4. A nodule activity promoter, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

5. A yield-increasing agent for leguminous plants contained in beans, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

6. A yield-increasing agent for leguminous plants contained in beans, wherein: The preparation is composed of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

7. The preparation according to any one of claims 1 to 6, wherein The ratio of (A) ascorbic acid or a salt thereof to (C) an organic solvent is, by mass ratio, 10 to 3,000,000 for the component (A) and 4 to 100,000 for the component (C), when the component (B) is 1.

8. The preparation according to any one of claims 1 to 7, wherein (B) The antioxidant further contains one or more selected from the group consisting of sodium sulfite, potassium sulfite, glutathione and uric acid.

9. The preparation according to any one of claims 1 to 8, wherein (C) The organic solvent is at least one selected from the group consisting of isobutyl alcohol and dimethyl sulfoxide.

10. The preparation according to any one of claims 1 to 9, wherein The composition is further combined with (D) a surfactant.

11. The preparation according to claim 10, wherein When the component (B) is 1, the ratio of the surfactant (D) is 10 to 300,000 in mass ratio.

12. The preparation according to claim 10 or 11, wherein (D) The surfactant includes a nonionic surfactant and / or an anionic surfactant.

13. The preparation according to claim 12, wherein (D) the nonionic surfactant is at least one selected from the group consisting of sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters and polyoxyethylene alkyl ethers, The anionic surfactant is an alkyl sulfate ester.

14. The preparation according to claim 12, wherein (D) The surfactant includes a nonionic surfactant and an anionic surfactant.

15. The preparation according to claim 14, wherein (D) the nonionic surfactant is at least one selected from the group consisting of sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters, and polyoxyethylene alkyl ethers, The anionic surfactant is one or more selected from alkyl sulfate ester salts and fatty acid salts.

16. The preparation according to any one of claims 1 to 15, wherein The composition is further combined with (E) a chelating agent.

17. The preparation according to claim 16, wherein The ratios of (A) ascorbic acid or a salt thereof, (C) an organic solvent, (D) a surfactant and (E) a chelating agent are calculated by mass ratio. When component (B) is set to 1, component (A) is 10 to 3,000,000, component (C) is 4 to 100,000, component (D) is 10 to 300,000 and component (E) is 0.01 to 100.

18. The preparation according to claim 16 or 17, wherein (E) The chelating agent is at least one selected from the group consisting of ethylenediaminetetraacetic acid and ethylenediamine-N,N′-disuccinic acid.

19. The preparation according to any one of claims 1 to 18, wherein It is also combined with a (F) vector.

20. The preparation of claim 19, wherein When the component (B) is 1, the ratio of the carrier (F) is 10 to 10,000 in terms of mass ratio.

21. The preparation according to claim 19 or 20, wherein (F) The carrier is at least one selected from the group consisting of zeolite, silica, bentonite and sodium sulfate.

22. The preparation according to any one of claims 1 to 9, wherein It is composed of a first agent containing the above-mentioned component (A) and a second agent containing the above-mentioned components (B) and (C), and the two are combined when used.

23. A method for promoting the growth of leguminous plants contained in beans, wherein: The method comprises the steps of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the resultant mixture to soil or plants. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

24. A method for promoting nodule activity, wherein: The method comprises the steps of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the resultant mixture to soil or plants. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

25. A method for increasing the yield of leguminous plants contained in beans, wherein: The method comprises the steps of combining (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent with water and applying the resultant mixture to soil or plants. (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

26. The method according to any one of claims 23 to 25, wherein: A first agent containing the component (A) and a second agent containing the components (B) and (C) are prepared, and when used, the two agents are applied in combination to soil or plants.

27. The formulation of claim 22, wherein When the second dose is set to 1, the weight ratio of the first dose is 1-100.

28. The method of claim 26, wherein: The first and second doses are dissolved in water for use.

29. The method of claim 28, wherein: The concentration of the first agent in the aqueous solution is 100 mass ppm to 300,000 mass ppm, and the concentration of the second agent is 10 mass ppm to 100,000 mass ppm.

30. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a growth promoter for leguminous plants contained in beans, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

31. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent for producing a growth promoter for leguminous plants contained in beans, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

32. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent in the manufacture of a nodule activity promoter, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

33. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant and (C) an organic solvent in the manufacture of a nodule activity promoter, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

34. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent for producing a yield-increasing agent for leguminous plants contained in beans, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 in mass ratio.

35. Use of a combination of (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent for producing a yield-increasing agent for leguminous plants contained in beans, wherein: (B) the antioxidant is at least one selected from the group consisting of tocopherol, butylated hydroxytoluene and butylated hydroxyanisole, (C) the organic solvent is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide, When the component (B) is 1, the ratio of the organic solvent (C) is 4 to 100,000 in mass ratio.

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