Plant growth promoter composition housed in container, and container housing same

Abstract

Provided are: a composition for promoting the growth of leguminous plants contained in beans, which is housed in a container; and a container for housing the composition. The present invention relates to a growth promoter composition for leguminous plants contained in legumes, the growth promoter composition being obtained by combining a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (B) an antioxidant and (C) an organic solvent, the antioxidant (B) being at least one selected from the group consisting of tocopherol, dibutylated hydroxytoluene and butylated hydroxyanisole; the (C) organic solvent is one or more selected from the group consisting of an alcohol having 5 or less carbon atoms and dimethyl sulfoxide, the proportion (mass ratio) of (A) ascorbic acid or a salt thereof is 10-3,000 when component (B) is 1, or the proportion (mass ratio) of (C) organic solvent is 4-100,000 when component (B) is 1, and the content (mass ratio) of (A) ascorbic acid or a salt thereof is 1. The first agent and the second agent are respectively accommodated in any one bottle body of a double-end type bottle body formed by connecting a first bottle body and a second bottle body through a connecting part.

Description

Technical Field

[0001] The present invention relates to a composition for promoting the growth of leguminous plants contained in beans, which is contained in a container, and the container containing the composition. Background Art

[0002] About one-third of the land on Earth is dry land, and due to future warming, it is expected that dry land will increase further. In addition, as a countermeasure to the serious food shortage caused by population growth, it is urgent to develop a technology to improve, maintain, and increase the yield of grains such as soybeans, rice, wheat, and corn in arid areas, salt accumulation areas, high temperature and low temperature areas, that is, areas where it is difficult to grow or growth deteriorates and yield decreases. In particular, soybeans are important grains and are widely consumed around the world, led by Japan. In addition, unlike other grains, soybeans have a high proportion of protein and lipids and are rich in nutritional value. Therefore, it is also important as a feed or oil raw material, and the development of technology to increase yield is underway.

[0003] Oxygen is an essential substance for many animals and plants to survive, but on the other hand, oxygen will produce highly reactive oxygen species in the cells of animals and plants, causing serious damage to the organisms, such as gene damage or enzyme inactivation. For this reason, various antioxidant substances including L-ascorbic acid are accumulated in plants, and a complex enzyme system that uses these antioxidant substances to eliminate reactive oxygen species works in various parts of the cells.

[0004] From this point of view, antioxidants containing ascorbic acid are often added to the raw materials applied to plants to promote growth. For example, non-patent document 1 discloses the effect of promoting growth or increasing yield by applying ascorbic acid to olives, sugar cane, wheat, etc. In addition, non-patent document 2 discloses that when a certain concentration of ascorbic acid aqueous solution dissolved in distilled water is sprayed on the leaves of leguminous crops for a total of three times during the vegetative growth period or the reproductive growth period, a maximum yield increase effect of 30% can be achieved. 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 glycyrrhizin 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 PotentialOxidantScavenger and Its Role in Plant Development and Abiotic StressTolerance. Frontiers in Plant Science, 8: 613)

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

[0009] Technical problem to be solved by the invention

[0010] The present invention relates to providing a composition for promoting the growth of legumes contained in beans, which is contained in a container, and a container for containing the composition.

[0011] Technical solutions for solving technical problems

[0012] The inventors of the present invention have studied the growth promotion of plants using ascorbic acid, and 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 promotion effect based on ascorbic acid is hardly achieved. It has also been found that by using ascorbic acid in combination with a specific antioxidant dissolved in an organic solvent, specifically, by containing them in separate containers and mixing them at the time of use, a root nodule activity promotion effect and a growth promotion effect can be achieved.

[0013] That is, the present invention is based on the following inventions 1) and 2).

[0014] 1) A composition, which is a growth promoter composition for leguminous plants contained in beans, comprising a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (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 5 or less carbon atoms and dimethyl sulfoxide, and the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 when the component (B) is 1, or the ratio (mass ratio) of (C) the organic solvent is 4 to 100,000 when the component (B) is 1, and the first agent and the second agent are respectively contained in one of the bottles of a double-headed bottle body formed by connecting the first bottle body and the second bottle body by a connecting member.

[0015] 2) A double-headed bottle, which is a bottle for containing the composition described in 1), wherein the wide-mouth bottle head of the second bottle having a bottle body and a wide-mouth bottle head is connected to the lower part of the first bottle having a bottle body with an external thread at the lower part, a narrow-mouth bottle head and a cover body via a connecting member.

[0016] Effects of the Invention

[0017] According to the present invention, the growth amount of the legumes contained in the beans is increased, the root nodule activity is promoted, and the yield of the fruit, seed, or fruit of the plant can be increased. In addition, according to the present invention, the first agent and the second agent with different functions are respectively contained in independent bottles with good sealing properties, and are sold as a whole during sales. When used, the consumer separates the two connected bottles, and can easily prepare a mixture of the first agent and the second agent mixed in a specified ratio. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is the seed weight per soybean plant.

[0019] Figure 2 The solubility when the mixing ratio of BHT and isobutyl alcohol is changed.

[0020] Figure 3 It is the solubility of a mixture of BHT and organic solvent (1:4).

[0021] Figure 4 It is the weight of soybean nodules one week after foliar spraying.

[0022] Figure 5 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0023] Figure 6 It is the amount of allantoin contained in soybean exudate one week after foliar spraying.

[0024] Figure 7 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0025] Figure 8 This is the soybean plant body in test plot 7 one week after foliar spraying.

[0026] Fig. 9 It is the seed weight per soybean plant.

[0027] Fig.10 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0028] Fig.11 It is the aboveground dry weight of adzuki beans one week after foliar spraying.

[0029] Fig.12 It is the dry weight of the underground part of adzuki beans one week after foliar spraying.

[0030] Fig.13 It is the aboveground dry weight of chickpea 1 week after foliar spraying.

[0031] Fig.14 It is the number of pods per individual of Lotus japonicus.

[0032] Fig.15 It is the seed weight of each individual of Lotus japonicus.

[0033] Fig.16 It is the number of beans per individual bean.

[0034] Fig.17 It is the weight of each bean fruit.

[0035] Fig.18 This is the appearance of the one-dose or two-dose composition after storage for one week.

[0036] Fig.19 It is the amount of allantoic acid contained in soybean exudate one week after foliar spraying.

[0037] Fig. 20 It is the dry weight of aboveground part of soybeans one week after foliar spraying.

[0038] Fig.21 It is the dry weight of underground parts of soybeans one week after foliar spraying.

[0039] Fig. 22 The diagram is a front view and a cross-sectional view showing a double-headed bottle body (a state in which a first bottle body and a second bottle body are connected) according to the first embodiment.

[0040] Fig.23 yes Fig. 22 An exploded perspective view of a double-headed bottle body is shown.

[0041] Fig.24 yes Fig.23 A perspective view and a cross-sectional view of the connecting parts shown.

[0042] Fig.25 It is a front view and a cross-sectional view showing a double-headed bottle body (a state where a first bottle body and a second bottle body are connected) according to a second embodiment.

[0043] Fig.26 yes Fig.25 An exploded perspective view of a double-headed bottle body is shown.

[0044] Fig. 27 yes Fig.26 A perspective view and a cross-sectional view of the connecting parts shown.

[0045] Fig.28 This is a perspective view of a double-headed bottle body according to the first embodiment. Left: a first bottle body and a second bottle body are connected, right: a first bottle body is separated.

[0046] Fig.29 This is a perspective view of a double-headed bottle body according to the second embodiment. Left: a first bottle body and a second bottle body are connected, right: a first bottle body is separated.

[0047] Fig.30 It is the soybean nodule activity one week after foliar spraying.

[0048] Fig.31 It is the dry weight of aboveground part of soybean 2 days after foliar spraying.

[0049] Description of Reference Numerals

[0050] 1: first bottle body; 2: second bottle body; 3: bottle body with external thread at the bottom; 4: narrow-mouth bottle head; 5: cover body; 6: bottle body; 7: wide-mouth bottle head; 8: connecting member; 9: cover plate; 10: first ring wall of connecting member 8; 11: second ring wall of connecting member 8; 12: external thread of bottle body 3; 13: screw structure of first bottle body and connecting member 8; 14: screw structure of second bottle body and connecting member 8; 15: internal thread of inner side wall of first ring wall 10 ; 16: external threaded portion of the wide-mouth bottle head 7; 17: internal threaded portion of the inner wall of the second ring wall 11; 18: connecting member; 19: cover plate; 20: first ring wall of the connecting member 18; 21: second ring wall of the connecting member 18; 22: internal threaded portion of the inner wall of the first ring wall 20; 23: screw-fitting structure of the first bottle body and the connecting member 18; 24: internal threaded portion of the inner wall of the second ring wall 21; 25: screw-fitting structure of the second bottle body and the connecting member 18; 26a, 26b double-headed bottle body. DETAILED DESCRIPTION

[0051] <Growth promoter composition for leguminous plants contained in beans>

[0052] In the growth promoter composition for legumes contained in beans of the present invention (hereinafter also referred to as "the composition of the present invention"), "growth promotion" means increasing the growth amount (fresh weight, height growth, etc.) of the legumes contained in the beans, and increasing the yield of fruits, seeds, or fruits.

[0053] In addition, when the plant is a nodulating plant, its growth promotion includes the promotion of nodule activity. Promoting nodule activity refers to promoting nodule activity, i.e., the nitrogen fixation function in the host plant exerted by nodules. Here, "nodule" refers to a nodule produced on the root of a plant through symbiosis with bacteria (rhizobium). Rhizobium reduces atmospheric nitrogen in nodules and converts it into ammoniacal nitrogen, which is supplied to the host in so-called symbiotic nitrogen fixation.

[0054] In the present invention, the Fabaceae plants included in the beans are so-called "beans" whose seeds and fruits are harvested and used, such as soybeans (including edamame), adzuki beans, chickpeas, lotus roots, kidney beans, peanuts, broad beans, peas, pinto 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, adzuki beans, kidney beans, peas, broad beans, chickpeas, and lotus roots, and more preferably soybeans.

[0055] The composition of the present invention is a growth promoter composition for leguminous plants contained in beans, which is a composition composed of a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (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 5 or less carbon atoms and dimethyl sulfoxide, and the ratio (mass ratio) of (A) ascorbic acid or a salt thereof is 10 to 3,000,000 when the component (B) is 1, or the ratio (mass ratio) of (C) the organic solvent is 4 to 100,000 when the component (B) is 1, and the first agent and the second agent are respectively contained in either bottle of a double-headed bottle body formed by connecting a first bottle body and a second bottle body by a connecting member. The composition is used by combining the two when used.

[0056] In the present invention, the IUPAC system name of ascorbic acid as component (A) 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 the L-form (so-called L-ascorbic acid) is preferred.

[0057] As ascorbic acid, commercially available products of various grades can be used.

[0058] Examples of the salt of ascorbic acid include sodium salt, potassium salt, calcium salt, magnesium salt, barium salt, ammonium salt, and salts with nitrogen-containing organic bases such as pyridine, trimethylamine, triethylamine, tributylamine, and diethylamine.

[0059] The concentration of (A) ascorbic acid or its salt in the composition when applying the composition of the present invention to plants can be appropriately adjusted in the range of, for example, 100 mass ppm or more and 300,000 mass ppm or less according to the supply method. For example, when spraying using a sprayer (e.g., a nozzle sprayer), it is preferably 100 mass ppm or more, more preferably 300 mass ppm or more, and further preferably 500 mass ppm or more in the spray liquid, and preferably 20,000 mass ppm or less, more preferably 10,000 mass ppm or less, and further 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 further preferably 500 to 4,500 mass ppm.

[0060] 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 even more 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 even more 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 even more preferably 150,000 to 200,000 mass ppm.

[0061] In the composition of the present invention, the antioxidant as 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.

[0062] Furthermore, in the present invention, as an antioxidant, antioxidants other than tocopherol, BHT and BHA, such as sodium isoascorbate, propyl gallate, sodium sulfite, potassium sulfite, potassium pyrosulfite, chlorogenic acid, catechin, glutathione, uric acid, etc. may also be used together, preferably one or more selected from sodium sulfite, potassium sulfite, potassium pyrosulfite, chlorogenic acid, catechin, glutathione and uric acid.

[0063] Therefore, as the antioxidant of component (B), one or more phenolic antioxidants selected from BHT and BHA are used. In addition to the phenolic antioxidants, as a preferred embodiment, one or more selected from sodium sulfite, potassium sulfite, glutathione and uric acid can be used in combination.

[0064] If ascorbic acid is dissolved in water containing metal ions such as tap water, the metal ions and ascorbic acid react to generate hydrogen peroxide, and then the hydrogen peroxide and the metal ions undergo a Fenton reaction to generate hydroxyl radicals. Furthermore, if hydroxyl radicals invade the plant body, a continuous lipid peroxidation reaction occurs that reacts with lipids present in the cell membrane, etc., generating lipid radicals or lipid peroxyl radicals. By using an antioxidant, this free radical can be supplemented.

[0065] In the present invention, by using the (B) antioxidant in a solution state dissolved in the (C) organic solvent, the plant growth promoting or nodule promoting activity effect of ascorbic acid can be effectively exerted.

[0066] In addition, by using a two-dose composition in which a preparation (first dose) containing component (A) of the present invention and a preparation (second dose) containing components (B) and (C) are separately prepared and combined, discoloration that occurs when the preparation containing all components (A), components (B) and (C) is stored can be suppressed.

[0067] Furthermore, the first agent containing component (A) may contain an antioxidant different from 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.

[0068] When the composition of the present invention is applied to plants, the concentration of the antioxidant (B) in the composition (mixture of the first agent and the second agent) 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.

[0069] The organic solvent as the component (C) is a solvent used to dissolve the antioxidant (B), and specifically is at least one selected from alcohols having 5 or less carbon atoms and dimethyl sulfoxide (DMSO).

[0070] Here, as the alcohol having 5 or less carbon atoms, preferably 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, glycerol, etc. are mentioned, and isobutanol and DMSO are more preferred.

[0071] When one or more selected from BHT and BHA are used as the (B) antioxidant, DMSO, isobutyl alcohol or a mixture thereof is preferably used as the (C) organic solvent.

[0072] When the composition of the present invention is applied to plants, the concentration of the organic solvent (C) in the composition (mixture of the first agent and the second agent) can be appropriately adjusted within a range of 1 ppm by mass to 100,000 ppm by mass, depending on the method of administration.

[0073] For example, when spraying is performed using a sprayer (e.g., a nozzle sprayer), the amount in the spray liquid is preferably 1 mass ppm or more, more preferably 10 mass ppm or more, and even more preferably 50 mass ppm or more, and is preferably 10,000 mass ppm or less, more preferably 5,000 mass ppm or less, and even more 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 even more preferably 50 to 1,000 mass ppm.

[0074] In the case of aerial spraying, the amount of 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.

[0075] Regarding 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 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 by a sprayer, it is preferably 500 to 45,000, and when spraying by aerial spraying, it is preferably 150,000 to 2,000,000. Furthermore, when spraying by a sprayer, it is more preferably 500 to 4,500, and when spraying by aerial spraying, it is preferably 150,000 to 200,000.

[0076] In addition, when component (B) is set to 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 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, it is preferably 20 to 10,000 when sprayed by a sprayer, and preferably 2,000 to 100,000 when sprayed by aerial spraying. Furthermore, it is more preferably 50 to 1,000 when sprayed by a sprayer, and more preferably 5,000 to 10,000 when sprayed by aerial spraying.

[0077] In the present invention, (D) a surfactant may be used in combination with (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent.

[0078] By using a surfactant, the wettability, adhesion, and permeability of (A) ascorbic acid or a salt thereof to the plant surface can be improved, and the effect of (A) ascorbic acid or a salt thereof can be enhanced or the effect can be efficiently exerted.

[0079] The (D) surfactant is preferably contained in the second agent containing the components (B) and (C).

[0080] As the surfactant, a nonionic surfactant and / or anionic surfactant may be contained.

[0081] Examples of the nonionic surfactant include at least one 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, polyoxyalkylene aryl ethers, polyoxyethylene alkenyl ethers, alkyl polyglycosides, polyoxyalkylene alkyl polyglycosides, and sucrose fatty acid esters. Examples of the anionic surfactant include at least one selected from alkyl sulfates, alkylbenzene sulfonates, polyoxyalkylene alkyl ether sulfates, polyoxyalkylene alkenyl ether sulfates, and polyoxyalkylene alkyl aryl ether sulfates, fatty acid salts, pyrophosphates, dodecyl phosphoric acid, polycarboxylic acid type polymers, polyoxyethylene alkylene alkyl acetic acid, aromatic sulfonate formalin condensates, polyoxyethylene distyrenated ether sulfates, alkyl diphenyl ether disulfonates, dialkyl sulfosuccinates, alkyl naphthalene sulfonates, and the like.

[0082] Among them, from the viewpoint of preventing phytotoxicity to plants during excessive application, the nonionic surfactant is preferably one or more selected from sorbitan fatty acid esters, glycerol fatty acid esters, polyoxyalkylene sorbitan fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters, polyoxyethylene alkyl ethers, alkyl polyglycosides and sucrose fatty acid esters, and more preferably one or more selected from 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 preferably one or more selected from alkyl sulfates (e.g., sodium lauryl sulfate, ammonium lauryl sulfate, triethanolamine lauryl sulfate, etc.) and fatty acid salts.

[0083] When the composition of the present invention is applied to plants, the concentration of the surfactant (D) in the composition (mixture of the first agent and the second agent) can be appropriately adjusted within the range of 10 mass ppm to 30,000 mass ppm depending on the application method.

[0084] For example, when spraying is performed using a sprayer (e.g., a nozzle sprayer), the amount 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.

[0085] In the case of aerial spraying, the amount of the chlorinated hydrocarbon in 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.

[0086] In addition, regarding the ratio (mass ratio) in the case of combining with the surfactant (D), 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 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, it is preferably 100 to 5,000 when sprayed by a sprayer, and preferably 1,000 to 50,000 when sprayed by aerial spraying. Moreover, it is more preferably 100 to 500 when sprayed by a sprayer, and more preferably 1,000 to 5,000 when sprayed by aerial spraying.

[0087] In the present invention, (E) a chelating agent may be used in combination with (A) ascorbic acid or a salt thereof, (B) an antioxidant, and (C) an organic solvent.

[0088] 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.

[0089] (E) The chelating agent is preferably contained in the first agent containing the component (A).

[0090] As the chelating agent, for example, aminocarboxylic acid chelating agents, phosphonic acid chelating agents, hydroxycarboxylic acid chelating agents, polycarboxylic acid chelating agents, etc. can be mentioned. As the aminocarboxylic acid chelating agent, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), methylglycinediacetic acid (MGDA), triethylenetetraaminehexaacetic acid (TTHA), glutamic aciddiacetic acid (GLDA), hydroxyethyliminodiacetic acid (HIDA), dihydroxyethylglycine (DHEG), aspartic aciddiacetic acid (ASDA), ethylenediaminesuccinic acid (EDDS) and salts thereof can be mentioned. As the phosphonic acid chelating agent, hydroxyethylidene diphosphonic acid (HEDP), nitrilotrimethylenephosphonic acid (NTMP), phosphonobutanetricarboxylic acid (PBTC), ethylenediaminetetramethylenephosphonic acid (EDTMP) and salts thereof can be mentioned.

[0091] Examples of the hydroxycarboxylic acid chelating agent include citric acid, malic acid, tartaric acid, gluconic acid, lactic acid, and salts thereof.

[0092] Examples of the polycarboxylic acid chelating agent include succinic acid, oxalic acid, glutaric acid, adipic acid, fumaric acid, malonic acid, and salts thereof.

[0093] When the composition of the present invention is applied to plants, the concentration of the chelating agent (E) in the composition (mixture of the first agent and the second agent) is preferably 0.01 mass ppm or more, more preferably 0.1 mass ppm or more, and even more preferably 1 mass ppm or more, and is preferably 100 mass ppm or less, more preferably 50 mass ppm or less, and even more 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 even more preferably 1 to 10 mass ppm.

[0094] In addition, regarding the ratio (mass ratio) in the case of combining with the chelating agent (E), when the component (B) is set to 1, the component (E) is preferably 0.01 or more, more preferably 0.1 or more, and even more preferably 1 or more, and is preferably 100 or less, more preferably 50 or less, and even more preferably 10 or less. In addition, it is preferably 0.01 to 100, more preferably 0.1 to 100, even more preferably 1 to 100, and even more preferably 1 to 10.

[0095] Preferred combinations of (B) an antioxidant, (C) an organic solvent, and (D) a surfactant used together with (A) ascorbic acid include, for example, the following combinations.

[0096] The component (B) is an antioxidant containing at least one selected from BHT and BHA, the component (C) is an organic solvent containing isobutyl alcohol or DMSO, the component (D) is a surfactant containing 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 further containing an alkyl sulfate as desired.

[0097] · Component (B) is an antioxidant containing BHT, sodium sulfite, glutathione and uric acid, component (C) is an organic solvent containing isobutyl alcohol or DMSO, component (D) is a combination of a surfactant containing one or more selected from sorbitan fatty acid esters, glycerol fatty acid esters, polyalkylene glycol fatty acid esters, polyoxyethylene resin acid esters and polyoxyethylene alkyl ethers, and further containing an alkyl sulfate as desired.

[0098] As shown in the examples described below, when soybean seeds as a nodulation plant are sown and then inoculated with rhizobia to cultivate soybeans, if an antioxidant such as BHT and an organic solvent (second agent) are combined and added to ascorbic acid (first agent), the nodule weight and nodule activity increase, and the formation of urea-based nodules can be promoted, indicating that the growth of the host plant is promoted.

[0099] Therefore, the combination of ascorbic acid (first agent) and a specific antioxidant and an organic solvent (second agent) is a composition for promoting root nodule activity because it promotes the growth of legumes contained in beans. Since the amount of urea derived from nodules is reported to be correlated with the seed yield (diagnostic method of total basal fertilizer nitrogen for soybeans in rotation fields, Ibaraki Prefecture Agricultural Comprehensive Center Agricultural Research Institute H18 main results; http: / / www.pref.ibaraki.jp / nourinsuisan / noken / seika / h18pdf / documents / 27.pdf), the composition of the present invention is also useful for increasing seed yield.

[0100] In addition, since the nitrogen fixation function in the host plant can be evaluated as urea production capacity or amide production capacity, the promoting effect of the composition of the present invention on nodule activity can be specifically evaluated as the urea production amount or amide production amount per wet nodule weight. The urea production amount or amide production amount can be measured and calculated in the case of cutting the plant body (for example, cutting the aerial part just below the cotyledon) in the exudate collected. Here, as urea, allantoin, allantoic acid or citrulline can be cited, and as amide, asparagine or glutamine can be cited, but it is preferred to measure the amount of allantoic acid or asparagine.

[0101] The composition obtained by combining the first agent and the second agent as described above promotes the growth or nodule activity of legumes contained in beans, and thus becomes a composition for increasing the yield of fruits, seeds or grains. For example, it can be a preparation for adding or formulating to various agricultural or horticultural materials, or to cultivation substrates for cultivating plants such as soil, culture medium, and nutrient solution.

[0102] The composition may be in the form of a liquid or gel composition, or in the form of a powder or granules.

[0103] Furthermore, the first and second doses are respectively contained in one of the two-headed bottles formed by connecting the first and second bottles via a connecting member.

[0104] The above composition may contain any component other than the above components (A) to (C) and (D) and (E). Such components include solvents (e.g., water, buffer, culture medium, nutrient 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, soybean meal, kaolin, montmorillonite, alumina, etc.), pH adjusters for promoting dissolution of the above compounds, developing agents for improving the developing power to plants or soil, fertilizer components for improving fertilizer efficiency, pesticide components, adhesives, and extenders. Agents, microorganisms that promote plant growth such as rhizobia or mycorrhizal fungi, essential nutrients for plants, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleic acid bases, sugars, monohydric alcohols, food additives, microbial extracts, plant hormones, nod factors, i.e., lipid chitosan oligosaccharides, synthetic lipid chitosan oligosaccharides, chitosan oligosaccharides, chitosan compounds, linoleic acid or its derivatives, linolenic acid or its derivatives, karrikin, acyl-homoserine lactone derivatives, betaine compounds, phenolic compounds, etc.

[0105] In addition, the composition for aerial spraying may contain an emulsifier such as polyoxyethylene fatty acid ester and an oil such as decanol.

[0106] As the form of the composition of the present invention, for example, there can be cited a cultivation substrate (such as a solution for agricultural or horticultural nutrient solution cultivation, etc.), a fertilizer, watering water, a microbial material such as a rhizobium material, a soil conditioner, a pesticide, a material for sowing, a plant supplement (such as an activator, a nutrient agent, etc.) containing the composition of the present invention, but it is not limited thereto. Among them, fertilizers, microbial materials, soil conditioners, materials for sowing, and plant supplements are preferred because they help improve the soil for cultivating plants. The fertilizer, microbial material, soil conditioner, material for sowing, and plant supplement may be solid or liquid, and in the case of solid, preferably powder 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 generally contain components of fertilizers, microbial materials, soil conditioners, materials for sowing, and plant supplements for cultivating plants.

[0107] The cultivation substrate, fertilizer, microbial materials such as rhizobium materials, soil conditioners, pesticides, materials for sowing, and plant supplements can also be prepared by adding the composition of the present invention to a common cultivation substrate (e.g., agricultural or horticultural soil, lodging, culture medium, nutrient solution, water, etc.), fertilizer, microbial materials such as rhizobium materials, soil conditioners, pesticides, materials for sowing, plant supplements (e.g., activators, nutrients, etc.), etc.

[0108] The composition of the present invention is used as a mixture of two bottles connected during use, wherein the first agent and the second agent are mixed at a predetermined ratio. The mass ratio of the first agent is 1 to 100, preferably 1 to 50, and more preferably 1 to 25, when the mass ratio of the second agent is 1.

[0109] 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.

[0110] The method for supplying the composition of the present invention is not particularly limited as long as the composition is applied to plants in a manner that can exert the effects of the present invention.

[0111] That is, as long as the composition (a mixture of the first agent and the second agent) comes into contact with the plant body of the legume plant contained in the beans or the soil in the rhizosphere of the plant, or is delivered to the plant body of the legume plant contained in the beans or the soil in the rhizosphere of the plant, there is no particular limitation, and examples include spraying on the soil surface, pouring, turning the soil, spraying on the leaves of plants, applying in admixture with fertilizers, adding to hydroponic solutions, or applying or smearing on seeds before sowing (for example, seed dressing), 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 preferred.

[0112] Furthermore, the spray liquid may be prepared at the time of application, and in this case, the dilution water used may be any of agricultural water, well water, groundwater, river water, lake water, tap water, and the like.

[0113] The spraying method is not particularly limited, and examples thereof include a spraying method, i.e., a method of spraying a spray liquid in a mist form by spraying. According to this method, the growth promoter or nodule activity promoter for leguminous plants contained in the beans of the present invention can exhibit good spreadability on the plants after being attached to the plants.

[0114] Specific examples of the spraying method include manual spraying using a mist blower, atomizer, sprayer (eg, pipe sprayer), etc., and aerial spraying using an airplane, helicopter, drone, etc.

[0115] The application amount of the composition of the present invention 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 application amount of component (A) 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 composition can be applied in the above range at one time or in multiple applications.

[0116] The time and frequency of application may also vary depending on the type of legume included in the beans. In the case of soybeans, when it is usually applied by spraying, pouring, turning the soil or mixing with seeds on the surface of a cultivation substrate such as soil, it is preferably applied once or 1 to 3 times before sowing or simultaneously with sowing. In addition, when applied after sowing, it is preferably applied between the early stage of the vegetative growth period before entering the reproductive growth period and the fruit enlargement period after entering the reproductive growth period.

[0117] <Double-headed bottle for storing the composition of the present invention>

[0118] In the composition of the present invention, the first agent and the second agent are respectively contained in one of the two-headed bottle bodies formed by connecting the first bottle body and the second bottle body via a connecting member.

[0119] The double-headed bottle of the present invention is a double-headed bottle formed by connecting a first bottle body and a second bottle body via a connecting member, wherein the first bottle body has a bottle body with an external thread at the bottom, a narrow-mouth bottle head and a cap, and the second bottle body has a bottle body and a wide-mouth bottle head. Figures 22-24 The double-headed bottle shown in the figure can be cited as a second embodiment. Figures 25-27 The double-headed bottle shown.

[0120] In either case, the first bottle body has a bottle body 3 having an external threaded portion at the bottom, a narrow-mouth bottle head 4, and a cap 5, and the second bottle body has a bottle body 6 and a wide-mouth bottle head 7 ( Fig.23 , Fig.26 The outer side walls of the narrow-mouth bottle head 4 of the first bottle body and the wide-mouth bottle head 7 of the second bottle body are respectively provided with external threaded parts for screwing with the cover body 5 or the connecting member 8 or 18.

[0121] <First Embodiment>

[0122] use Figures 22-24 A double-headed bottle having the connecting member 8 will be described.

[0123] The connecting member 8 includes a cover plate 9, a first ring wall 10 extending vertically upward along the outer periphery of the cover plate 9, and a second ring wall 11 formed by the first ring wall 10 extending horizontally outward and then folding vertically downward. Fig.24 An internal thread portion 15 is provided on the inner side wall of the first annular wall 10. In addition, an internal thread portion 17 ( Fig.24 ).

[0124] The external threaded portion of the first bottle body 1, which is provided at the lower part of the bottle main body 3 having the external threaded portion at the lower part, is connected to the first ring wall 10 of the connecting member 8 via a screw structure 13, and the second ring wall 11 of the connecting member 8 is connected to the wide-mouth bottle head 7 of the second bottle body 2 via a screw structure 14 ( Fig.23 ).

[0125] The screw structure 13 is composed of an external threaded portion 12 of the bottle main body 3 having an external threaded portion at the bottom of the first bottle body 1 and an internal threaded portion 15 provided on the inner side wall of the first ring wall 10. In addition, the screw structure 14 is composed of an external threaded portion 16 provided on the outer side wall of the wide-mouth bottle head 7 of the second bottle body 2 and an internal threaded portion 17 provided on the inner side wall of the second ring wall 11 ( Fig.23).

[0126] That is, the internal thread portion 15 of the first ring wall 10 is screwed with the external thread portion 12 of the bottle main body 3 having an external thread portion at the lower portion of the first bottle body 1, and the internal thread portion 17 of the second ring wall 11 is screwed with the external thread portion 16 of the outer wall of the second bottle body 2 provided on the wide-mouth bottle head 7 ( Fig.23 ).

[0127] The screwing direction of the screwing structure 13 between the first bottle body 1 and the connecting member 8 may be the same as or different from the screwing direction of the screwing structure 14 between the second bottle body 2 and the connecting member 8 .

[0128] <Second Embodiment>

[0129] use Figures 25-27 A double-headed bottle including the connecting member 18 will be described.

[0130] The connecting member 18 includes a cover plate 19, a first annular wall 20 and a second annular wall 21 which protrude vertically toward the outer periphery of the cover plate 19. Fig. 27 An internal thread portion 22 is provided on the inner side wall of the first annular wall 20. In addition, an internal thread portion 24 is provided on the inner side wall of the second annular wall 21 ( Fig. 27 ).

[0131] The external threaded portion of the first bottle body 1, which is provided at the lower part of the bottle main body 3 having an external threaded portion at the lower part, is connected to the first ring wall 20 of the connecting member 18 via a screw structure 23, and the connecting member 18 is connected to the wide-mouth bottle head 7 of the second bottle body 2 via a screw structure 25 ( Fig.26 ).

[0132] The screw structure 23 is composed of the external threaded portion 12 of the bottle main body 3 having an external threaded portion at the bottom of the first bottle body 1, and the internal threaded portion 22 provided on the inner side wall of the first ring wall 20. In addition, the screw structure 25 is composed of the external threaded portion 16 provided on the outer side wall of the wide-mouth bottle head 7 of the second bottle body 2, and the internal threaded portion 24 provided on the inner side wall of the second ring wall 21 ( Fig.26 ).

[0133] That is, the internal thread portion 22 of the first annular wall 20 is screwed with the external thread portion 12 of the bottle main body 3 having an external thread portion at the lower portion of the first bottle body 1, and the internal thread portion 24 of the second annular wall 21 is screwed with the external thread portion 16 of the outer wall of the second bottle body 2 provided on the wide-mouth bottle head 7 ( Fig.26 ).

[0134] The screwing direction between the first bottle body 1 and the connecting member 18 may be the same as or different from the screwing direction between the second bottle body 2 and the connecting member 18 .

[0135] The material of the first bottle body and the second bottle body may be synthetic resin or metal. As the synthetic resin, for example, polyethylene such as polyethylene terephthalate, polypropylene, high-density polyethylene, vinyl chloride, etc. may be cited. As the metal, aluminum or tinplate may be cited. The synthetic resin may be transparent or opaque.

[0136] The first bottle body and the second bottle body made of synthetic resin or metal can be manufactured by synthetic resin injection molding or synthetic resin blow molding or metal press molding using a mold.

[0137] The bottle body having an external threaded portion at the lower portion and the narrow-mouthed bottle head of the first bottle body and the bottle body and the wide-mouthed bottle head of the second bottle body are preferably integral products made by blow molding of synthetic resin.

[0138] The cover body connected to the narrow-mouth bottle head of the first bottle body is preferably made by injection molding of synthetic resin.

[0139] The material of the connecting member may be any of the materials cited as the materials of the first bottle body and the second bottle body. The material of the connecting member may also be the same material as the bottle body. From the viewpoint of the formability and handling of the spiral strip, the material of the connecting member is preferably a synthetic resin. When the material of the connecting member is a synthetic resin, it can be manufactured by injection molding using a mold. The connecting member is preferably an integral product made of a synthetic resin.

[0140] The first bottle and the second bottle contain separately a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (B) an antioxidant and (C) an organic solvent. The first agent and the second agent can be contained in either the first bottle or the second bottle.

[0141] The first agent containing (A) ascorbic acid or a salt thereof may be contained in a powdery or granular form. In this case, tap water, agricultural water, well water, etc. may be added to the bottle containing the first agent to prepare a liquid agent.

[0142] The first bottle body and the second bottle body are separated during use ( Fig.28 , Fig.29 ), the first agent and the second agent are mixed. For example, this method includes placing the composition in the first bottle into the second bottle, or placing the composition in the first bottle and the composition in the second bottle into another container respectively and mixing them.

[0143] For example, one method is to store a powdered first agent in a second bottle and a liquid second agent in a first bottle. Then, the liquid second agent in the first bottle is dissolved in tap water, agricultural water, well water, etc. that has been added to another container in advance, and then the powdered first agent in the second bottle is dissolved in the other container, or the first agent and the second agent are dissolved in the reverse order to prepare a liquid agent.

[0144] Regarding the above-mentioned embodiment, the present invention further discloses the following aspects.

[0145] <1> A composition is a growth promoter composition for leguminous plants contained in beans, wherein the composition is a composition comprising a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (B) an antioxidant and (C) an organic solvent,

[0146] (B) the antioxidant is at least one selected from 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, the ratio (mass ratio) of (A) ascorbic acid or its salt is 10 to 3,000,000 when component (B) is set to 1, or the ratio (mass ratio) of (C) the organic solvent is 4 to 100,000 when component (B) is set to 1, and the first agent and the second agent are respectively contained in any one of the double-headed bottles formed by connecting the first bottle body and the second bottle body by a connecting member.

[0147] <2> like <1> The composition, wherein

[0148] The first bottle body includes a bottle body having an external threaded portion at the lower portion, a narrow-mouth bottle head portion, and a cap body, and the second bottle body includes a bottle body and a wide-mouth bottle head portion.

[0149] <3> like <1> or <2> The composition, wherein

[0150] The first dose is contained in the second bottle, and the second dose is contained in the first bottle.

[0151] <4> like <1> ~ <3> The composition described in any one of the foregoing, wherein the ratio (mass ratio) of (A) ascorbic acid or a salt thereof to (C) an organic solvent is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, and the ratio (mass ratio) of component (A) to 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, when component (B) is set to 1.

[0152] <5> like <1> ~ <3> The composition described in any one of the preceding claims, wherein

[0153] (B) The antioxidant further comprises at least one selected from the group consisting of sodium sulfite, potassium sulfite, glutathione and uric acid.

[0154] <6> like <1> ~ <3> The composition described in any one of the preceding claims, wherein

[0155] (C) The organic solvent is at least one selected from isobutyl alcohol and dimethyl sulfoxide.

[0156] <7> like <1> ~ <6> Any one of the agents, wherein

[0157] The second agent further contains (D) a surfactant.

[0158] <8> like <7> The composition, wherein

[0159] (D) When the ratio (mass ratio) of the surfactant is 1, it is preferably 10 to 300,000, more preferably 50 to 100,000, and even more preferably 100 to 5,000.

[0160] <9> like <7> or <8> The composition, wherein

[0161] (D) The surfactant includes a nonionic surfactant and / or an anionic surfactant.

[0162] <10> like <9> The composition, wherein

[0163] (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, and the anionic surfactant is an alkyl sulfate ester salt.

[0164] <11> like <9> The agent, wherein

[0165] (D) The surfactant includes a nonionic surfactant and an anionic surfactant.

[0166] <12> like <11> The composition, wherein

[0167] (D) The nonionic surfactant is one or more 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 one or more selected from alkyl sulfates and fatty acid salts.

[0168] <13> like <1> ~ <12> The composition described in any one of the preceding claims, wherein

[0169] (E) A chelating agent is also combined in the first dose.

[0170] <14> like <13> The composition, wherein

[0171] When the ratio (mass ratio) of (A) ascorbic acid or a salt thereof, (C) an organic solvent, (D) a surfactant and (E) a chelating agent is 1, the ratio (mass ratio) of the component (A) is 10 to 3,000,000, preferably 100 to 2,000,000, more preferably 500 to 200,000, the ratio (C) is 4 to 100,000, preferably 10 to 50,000, more preferably 20 to 20,000, further preferably 50 to 10,000, the ratio (D) is preferably 10 to 300,000, more preferably 50 to 100,000, further preferably 100 to 5,000, and the ratio (E) is preferably 0.01 to 100, more preferably 0.1 to 100, further preferably 1 to 10, further preferably 1 to 10.

[0172] <15> like <13> or <14> The composition, wherein

[0173] (E) The chelating agent is at least one selected from ethylenediaminetetraacetic acid and ethylenediamine-N,N'-disuccinic acid.

[0174] <16> exist <1> ~ <15> The legumes included in the beans are preferably beans selected from the group consisting of soybeans, adzuki beans, chickpeas, lotus roots, kidney beans, peanuts, broad beans, peas, pinto beans, lima beans, mung beans, cowpeas, lentils and sword beans, more preferably beans selected from the group consisting of soybeans, adzuki beans, kidney beans, peas, broad beans, chickpeas and lotus roots, and further preferably soybeans.

[0175] <17> A double-headed bottle body, which is used to contain <1> The bottle of the composition, wherein the first bottle has: a bottle body with an external thread at the bottom; a narrow-mouth bottle head; and a cap, and the second bottle has a bottle body and a wide-mouth bottle head. The wide-mouth bottle head of the second bottle is connected to the bottom of the first bottle via a connecting member.

[0176] <18> like <17> The bottle body, wherein

[0177] The connecting component comprises: a cover plate; a first annular wall extending vertically upward along the outer peripheral edge of the cover plate; and a second annular wall formed by the first annular wall extending horizontally outward and then folding vertically downward, the external threaded portion of the first bottle body provided at the lower part of the bottle main body having an external threaded portion at the lower part is screwed with the internal threaded portion of the connecting component provided on the inner side wall of the first annular wall, and the external threaded portion of the wide-mouth bottle head provided on the second bottle body 2 is screwed with the internal threaded portion of the connecting component provided on the inner side wall of the second annular wall, thereby connecting the first bottle body and the second bottle body.

[0178] <19> like <17> The bottle body, wherein

[0179] The connecting component has a cover plate and a first annular wall and a second annular wall protruding in a vertical direction toward the outer peripheral edge of the cover plate, respectively; an external threaded portion of the first bottle body arranged at the lower part of the bottle main body having an external threaded portion at the lower part is screwed with an internal threaded portion of the connecting component arranged on the inner side wall of the first annular wall; an external threaded portion of the wide-mouth bottle head arranged on the second bottle body is screwed with an internal threaded portion of the connecting component arranged on the inner side wall of the second annular wall, thereby connecting the first bottle body and the second bottle body.

[0180] Example

[0181] Test Example 1 Effect of ascorbic acid dissolved water on application effect

[0182] 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 ascorbic acid-dissolved water on the application effect was examined in soybean (Glycine max).

[0183] The effect of 500 mass ppm of ascorbic acid applied once to the leaves on soybean yield was evaluated. The evaluated test plots 1 to 3 are as follows.

[0184] 1st division: control (no application)

[0185] 2 divisions: 500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0186] 3rd division: 500 ppm by mass of ascorbic acid for foliar spraying (dissolved in deionized water)

[0187] (1) Cultivation conditions

[0188] Soybean cultivation was carried out at a farm in Tochigi Prefecture, and the soybean cultivar used was "Satonohohoemi". In the farm, foliar spraying was performed with the width of the ridge excluding the peripheral part as 2m as one division. For one test plot, the control (test plot 1) was tested in 12 divisions, and the ascorbic acid application area (test plots 2 and 3) was tested in 3 divisions. An average of 19 individuals were cultivated within the range of 2m set as one division.

[0189] (2) Foliar spraying and determination of seed weight

[0190] As ascorbic acid, "Food Additive Grade Vitamin C (L-Ascorbic Acid) Fine Mesh Type SSS" manufactured by Fuso Chemical Industry Co., Ltd. was used. It is made by dissolving a 500 mass ppm aqueous solution of ascorbic acid in tap water or deionized water. Using a dry cell sprayer (GT-5HS, Koshin Co., Ltd.), 20 mL of ascorbic acid aqueous solution was sprayed on the entire soybean plant body for each plant. Spraying was carried out on the 60th day after sowing, and the growth stage was a period corresponding to the fruit enlargement period. Harvesting was carried out on the 108th day after sowing. Whole fruits were recovered from each individual after harvesting and dried at 100°C for 48 hours. As yield data, the mass of dried fruits was measured.

[0191] (3) Results

[0192] After calculating the average fruit weight of each individual in the first division, the average and standard deviation of the three divisions were calculated ( Figure 1 ). The graph in the figure represents the mean ± standard deviation. When the same concentration of ascorbic acid is sprayed on the leaves, it is shown that the effect of tap water and deionized water on seed weight is different. That is, the technical problem that the effect of ascorbic acid disappears when water (tap water) used in agricultural sites is used becomes clear.

[0193] Test Example 2 Solubility of Butylated Hydroxytoluene in Various Solvents

[0194] The amount of BHT dissolved in 100 μL of isobutyl alcohol was varied. It was confirmed that BHT was uniformly dissolved to 25 mg, i.e., the ratio of BHT:isobutyl alcohol = 1:4 ( Figure 2 ). The solubility of other solvents was also confirmed at a ratio of BHT:solvent = 1:4. Among the solvents studied, it was uniformly soluble in ethanol, methanol, 2-propanol, 1-butanol, and dimethyl sulfoxide (DMSO) except glycerol ( Figure 3 ). BHT is known to be poorly soluble in water. In addition, each reagent used was manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.

[0195] Example 1 Preparation of a spray liquid having plant growth promoting, nodule promoting and income increasing effects

[0196] (1) Preparation of 2-dosage composition

[0197] The following ingredients were used to prepare the two-dose composition shown in Table 1. One or more selected from BHT, BHA and tocopherol as component (B) was pre-dissolved in component (C), and then component (D) was further 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 as 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.

[0198] (2) Preparation of spray solution and solubility evaluation

[0199] The prepared two-dose 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 dose was dissolved in heated tap water, the first dose was further dissolved, and the solubility was evaluated.

[0200] (3) Reagents used

[0201] Ascorbic acid used was "Food Additive Grade Vitamin C (L-Ascorbic Acid) Fine Mesh Type SSS" manufactured by Fuso Chemical Industries, Ltd. Sodium ascorbate, tocopherol, BHT, BHA, ethanol, methanol, 2-propanol, isobutanol, 1-butanol, DMSO, and citric acid used were manufactured by Fujifilm Wako Pure Chemical Industries, Ltd. EDTA used was manufactured by Dojindo 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.

[0202] (4) Results of solubility evaluation

[0203] The appearance of the spray liquid prepared using each composition was evaluated for solubility in aqueous solution according to the following criteria. The results are shown in Table 2.

[0204] ○: No dissolved residue, a uniform aqueous solution.

[0205] △: A small amount of dissolved residue was observed, but the mixture became uniform immediately after stirring.

[0206] ×: Dissolved residues were visible and the mixture was not uniform immediately after stirring.

[0207] The results in Table 2 show that in Composition Examples 1 to 17, uniform spray liquids suitable for spraying as plant growth promoters can be obtained.

[0208] Table 1

[0209]

[0210] Table 2

[0211]

[0212] Example 2 Evaluation of nodule weight and nodule activity

[0213] (1) Soil preparation and sowing

[0214] The mid-term fertilizer type soil (Takii water-containing cell soil mid-term fertilizer type, Takii Seedling Co., Ltd.) and vermiculite fine particles (Akagi Horticulture Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a polyethylene pot (diameter 10.5 cm, height 9 cm). Soybean seeds used "エンレイ" (purchased from Nikko Seedling Co., Ltd.). After 250 mL of tap water was given to each pot, 2 seeds were sown in each pot, and they were sown one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions in each test area was set to 6 (n = 6).

[0215] (2) Inoculation of Rhizobia

[0216] 1.5% agar (Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH 6.8)) to prepare a solid medium, and Bradyrhizobium japonicum NBRC14783T strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0217] (3) Cultivation conditions

[0218] Cultivation from sowing to thinning was carried out using an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m 2 / s) 16 hours / dark period 8 hours, the temperature was set to 30°C in the light period / 25°C in the dark period, and the humidity was set to 50%. After 7 days of sowing, the seedlings were thinned out in a manner of one plant per pot. After thinning, the seedlings were cultivated outdoors. After the water in the trough under the pot was gone, tap water was added to the trough to overflow the lower part of the pot by about 5 cm to water it.

[0219] (4) Foliar spraying

[0220] The spray liquid shown in Table 3 was prepared, and 6.7 mL was sprayed per plant using a mist blower on the 14th day after sowing. Foliar spraying was performed only once, and 7 test plots were studied.

[0221] Regarding the preparation of the spray liquid, BHT in component (B) is pre-dissolved in DMSO of component (C), and then component (C) and component (D) are mixed to prepare the second dose. Next, component (A), uric acid (antioxidant) and component (E) are mixed to prepare the first dose. After the second dose is dissolved in water, the spray liquid is prepared by further dissolving the first dose.

[0222] Here, ascorbic acid used is "Food Additive Grade Vitamin C (L-Ascorbic Acid) Pore Type SSS" manufactured by Fuso Chemical Industry Co., Ltd. BHT, uric acid, DMSO and EDTA used are manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., and EDDS used is "CHELEST EDDS-35" manufactured by CHELEST Corporation. As component (C), DMSO and isobutyl alcohol are used. In component (D), as a nonionic surfactant, sorbitan monolaurate or sorbitan fatty acid ester, polyoxyethylene resin acid ester is used. In addition, in component (D), fatty acid soap potassium salt is used as an anionic surfactant.

[0223] Table 3

[0224]

[0225] (5) Method for recovering exudate for measuring the amount of ureide nitrogen as an indicator of nodule activity

[0226] After 21 days of cultivation, the soybean stems were cut at the cotyledonary node using flower and tree scissors, and a 1.5 mL microtube (Eppendorf Corporation) plugged with a cotton ball #10 (Kawamoto Sangyo Co., Ltd.) was placed on the cap to collect the exudate flowing out from the cut surface of the stem for 2 hours. The cotton ball after collecting the exudate was stored in a freezer at -80°C. In addition, the amount of exudate was calculated by comparing the weight of the cotton ball before and after collecting the exudate.

[0227] (6) Extraction of exudate components recovered from cotton balls

[0228] The exudate components contained in the cotton ball were eluted with ultrapure water and filtered using a microbial spin chromatography column (Bio-Rad Laboratories Co., Ltd.). The filtrate remaining in the column was centrifuged and recovered by running a centrifuge (CR15RN, Hitachi, Ltd.) at 15,000 rpm for 1 minute. The filtrate diluted 1,000 times was quantified.

[0229] (7) Apparatus and measurement method used for quantification of ureide nitrogen content

[0230] The HPLC device and mass spectrometer used Agilent 1260 Infinity LC system (Agilent Technologies Co., Ltd.) and AB SCIEX TripleQuad 4500 system (AB Sciex Co., Ltd.), respectively. The chromatographic column used Scherzo SS-C18 (100 mm × 2 mm, 3 μm) (Imtakt Co., Ltd.), and the chamber temperature was set to 40°C. 5 μL of appropriately diluted sample was injected, the flow rate was 0.5 mL / min, and the eluent used 0.1% formic acid aqueous solution (eluent A) and 50 mM ammonium acetate / methanol solution (eluent B). After balancing with eluent A: eluent B = 95:5, the sample was supplied, and after 5 minutes, the ureide nitrogen was eluted by a linear gradient of eluent A: eluent B = 80:20.

[0231] (8) Standards used and quantification

[0232] The amount of ureide 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.

[0233] For each sample, allantoin and allantoic acid were identified based on the retention time, precision mass, and MS / MS spectrum of 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.

[0234] (9) Results

[0235] The results of nodule weight and nodule activity are shown in Figure 4 and Figure 5 The mean and standard deviation were calculated from the data of 4 replicates excluding the highest and lowest values ​​in each test area with 6 replicates. The graphs in the figures represent the mean ± standard deviation.

[0236] In Test Example 1, the effects of different water qualities for dissolving ascorbic acid on soybean yield are shown. According to the test plots 1 to 3 of this example, it can be confirmed that even in the early stage of soybean growth, when ascorbic acid is dissolved in deionized water, there is a trend of increasing nodule weight and nodule activity, and when it is dissolved in tap water, there is a trend of decreasing the application effect of ascorbic acid. The comparison between test plots 4 and 5 shows that in order to restore the nodule activity reduced by dissolving ascorbic acid in tap water, it is necessary to add an organic solvent. In addition, the comparison between test plots 6 and 7 shows that in order to restore the nodule activity, EDTA is more preferred among chelating agents.

[0237] Example 3 Study on the Mixing Ratio of Ascorbic Acid and Antioxidants

[0238] The preparation and sowing of soil, inoculation of rhizobia, the method for recovering the exudate for measuring the amount of ureide nitrogen which is an indicator of nodule activity, the experimental operations before quantifying the exudate, the apparatus and measurement method used in quantifying the amount of ureide nitrogen, the standards used and the items quantified were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0239] (1) Cultivation conditions

[0240] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (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 of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to cover the lower part of the pot by about 5 cm.

[0241] (2) Foliar spraying

[0242] As in Example 2, the spray solution shown in Table 4 below was prepared, and 6.7 mL was sprayed per plant using a mist blower on the 14th day after sowing. Foliar spraying was performed only once, and 7 test plots were studied (Table 4). The amount of water sprayed was constant, and the concentration of ascorbic acid in the spray solution was adjusted by changing the amount of ascorbic acid per plant. The reagents used were the same as in Example 2.

[0243] Table 4

[0244]

[0245] (3) Results

[0246] The results of the nodulation activity assay are shown in Figure 6 and Figure 7 Graphs in the figures represent mean ± standard deviation.

[0247] exist Figure 6 and Figure 7 In the experiment, the nodule activity when ascorbic acid dissolved in tap water was sprayed on the leaves did not change, but decreased, compared with the case where no application was performed. In test area 3, which was a composition without ascorbic acid, the nodule activity decreased, but the nodule activity was restored or improved by adding ascorbic acid to the composition of test area 3. In the experiment using the water volume assumed to be sprayed using a sprayer (such as a nozzle sprayer), the following trend was shown: the nodule activity in the test area 4 with ascorbic acid:BHT=500:1 was the highest, and the nodule activity decreased in test area 7 with ascorbic acid:BHT=20,000:1. It was also shown that: because ascorbic acid:BHT=20,000:1 Figure 8 The results showed that the leaves lost their color, so the mixing ratio of ascorbic acid, component (B), and component (C) was preferably 100 to 20,000:1:2 to 100. It was also shown that component (C) had a root nodule activity recovery effect when isobutyl alcohol was used.

[0248] Example 4: Effect of increasing soybean income on farms

[0249] The effect of foliar spraying of a composition containing 1,500 ppm of ascorbic acid on soybean yield was evaluated. Foliar spraying was performed only once, and eight test plots were studied (Table 5).

[0250] Table 5

[0251]

[0252] (1) Cultivation conditions

[0253] Soybean cultivation was carried out at a farm in Tochigi Prefecture, and the soybean cultivar used was "Satonohohoemi". In the farm, foliar spraying was performed with the width of the ridge excluding the periphery as 2m as one section. Three sections were tested for one test area. An average of 10 individuals were cultivated within the 2m range set as one section.

[0254] (2) Foliar spraying and determination of seed weight

[0255] 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 manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.

[0256] The spray solution was prepared by dissolving the reagent in tap water or deionized water. A dry cell sprayer (GT-5HS, Koshin Co., Ltd.) was used to spray the soybean plant as a whole with 6.7 mL of ascorbic acid aqueous solution for each plant. Spraying was performed on the 75th day after sowing, and the growth stage was a period equivalent to the fruit enlargement period. Harvesting was performed on the 173rd day after sowing. Whole fruits were recovered from each individual after harvesting and dried at 100°C for 48 hours. The mass of dried fruits was measured as yield data.

[0257] (3) Results

[0258] After calculating the average fruit weight of each individual in one zone, the average and standard deviation of the eight zones were calculated ( Fig. 9 ). Graphs in the figures represent mean ± standard deviation.

[0259] When the same concentration of ascorbic acid was sprayed on the leaves, it was confirmed again that tap water and deionized water had different effects on seed weight. This showed that by adding component (B), component (C) and component (D) to ascorbic acid, a yield-increasing effect could be obtained even when tap water was used. It was also shown that component (D) also had a yield-increasing effect when polyoxyethylene lauryl ether was used.

[0260] Example 5 Evaluation of the nodulation activity of high concentration application of ascorbic acid

[0261] The preparation and sowing of soil, inoculation of rhizobia, cultivation conditions, method of recovering exudate for measuring the amount of ureide nitrogen which is an indicator of nodule activity, experimental operations before quantifying the exudate, apparatus and determination method used in quantifying the amount of ureide nitrogen, standard substances used and items to be quantified were the same as those in Example 3, and soybean cultivation was carried out for 21 days.

[0262] (1) Foliar spraying

[0263] Foliar spraying was performed 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 nonionic surfactants. The spray solution was prepared by dissolving the reagents in tap water or deionized water. Foliar spraying was performed 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 mist blower, and test areas 4 to 8 were sprayed with 0.125 mL per plant.

[0264] Table 6

[0265]

[0266] (2) Results

[0267] The results of the nodulation activity assay are shown in Fig.10 The mean and standard deviation were calculated from the data of 4 replicates excluding the highest and lowest values ​​in each test area with 6 replicates. The graphs in the figures represent the mean ± standard deviation.

[0268] exist Fig.10 In the experiment, a trend of increasing nodule activity of soybean can be seen in the experiment areas 4 and 5 where ascorbic acid was sprayed with a small amount of water compared to the experiment areas 2 and 3. Regardless of the amount of water used for spraying, the nodule activity was improved when deionized water was used for dissolving ascorbic acid compared to the case of using tap water. According to the comparison between the experiment areas 5 and 6, the nodule activity was improved to the greatest extent by adding BHT as compound (B), DMSO as compound (C), and polyethylene glycol monolaurate as compound (D) to ascorbic acid. On the other hand, it was shown that in the experiment area 7, the nodule activity decreased when the ascorbic acid concentration was increased to 300,000 ppm compared to the experiment area 6, and therefore, when spraying with a small amount of water, the ascorbic acid concentration is preferably 300,000 ppm or less. In addition, according to the comparison between the experiment areas 8 and 6, DMSO in the compound (C) is more preferred than isobutanol. It was also shown that when polyethylene glycol monolaurate was used as compound (D), it also had the effect of restoring root nodule activity.

[0269] Example 6 Evaluation of initial growth indicators in adzuki beans

[0270] (1) Soil preparation and sowing

[0271] The mid-term fertilizer type soil (Takii water-containing cell soil mid-term fertilizer type, Takii Seedling Co., Ltd.) and vermiculite fine particles (Akagi Horticulture Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a polyethylene pot (diameter 10.5 cm, height 9 cm). The adzuki bean seeds used "Tamba Dainagon Azuki Bean" (purchased from Takii Seedling Co., Ltd.). After 250 mL of tap water was given to each pot, 2 seeds were sown in each pot, and one seed was sown at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions in each test area was set to 6 (n = 6).

[0272] (2) Inoculation of Rhizobia

[0273] 1.5% agar (Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH 6.8)) to prepare a solid medium, and Bradyrhizobium japonicum NBRC14783T strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0274] (3) Cultivation conditions

[0275] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m 2 / s) 16 hours in the light period / 8 hours in the dark period, the temperature was set to 25°C in the light period / 20°C in the dark period, and the humidity was set to 50%. After 7 days of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to cover the lower part of the pot by about 5 cm.

[0276] (4) Foliar spraying

[0277] The spray solution was prepared and sprayed once with 6.7 mL per plant using a mist blower on the 21st day after sowing. For the preparation of the spray solution, BHT was dissolved in isobutyl alcohol, and then isobutyl alcohol and sorbitan monolaurate were mixed and dissolved in water. Thereafter, ascorbic acid was dissolved in water to prepare the solution. The reagents used were the same as those in Example 2. The test plots 1 to 4 evaluated were as follows.

[0278] On the 28th day after sowing, the plants were dried at 90°C for 24 hours, and then the above-ground dry weight and below-ground dry weight of the plants were measured.

[0279] 1st division: control (no application)

[0280] 2-zone: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0281] 3rd division: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in deionized water)

[0282] 4 divisions: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + isobutyl alcohol 50 mass ppm + sorbitan monolaurate 350 mass ppm foliar spray (dissolved in tap water)

[0283] (5) Results

[0284] The aboveground dry weight and underground dry weight are shown in Fig.11 and Fig.12 Graphs in the figures represent mean ± standard deviation.

[0285] According to Test Example 1 and Example 2, it was shown that in soybeans, when the same concentration of ascorbic acid was sprayed on the leaves, tap water and deionized water had different effects on seed weight and initial growth indicators. It was shown that in adzuki beans, as in soybeans, the effect on initial growth indicators was different depending on the quality of the water in which ascorbic acid was dissolved. According to the comparison between Test Areas 2 and 4, it was shown that by adding component (B), component (C) and component (D) to ascorbic acid, a growth-promoting effect could be obtained even when tap water was used.

[0286] Example 7 Evaluation of initial growth indicators in chickpea

[0287] (1) Soil preparation and sowing

[0288] The mid-term fertilizer type soil (Takii water-containing cell soil mid-term fertilizer type, Takii Seedling Co., Ltd.) and vermiculite fine particles (akagi Horticulture Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a polyethylene pot (diameter 10.5cm, height 9cm). Chickpeas used seeds of Kabuli (Kabuli) species (purchased from Nikko Seedling Co., Ltd.). After 250mL of tap water was given to each pot, 2 seeds were sown in each pot, and one seed was sown at a depth of about 1 to 2cm from the soil surface. In addition, the number of repetitions in each test area was set to 6 (n = 6).

[0289] (2) Inoculation of Rhizobia

[0290] 1.5% agar (Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH 6.8)) to prepare a solid medium, and Mesorhizobium ciceri NBRC100389T strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0291] (3) Foliar spraying

[0292] The cultivation conditions and the method of foliar spraying were the same as those in Example 6 (the example of adzuki beans). The test plots 1 to 4 evaluated were as follows.

[0293] On the 28th day after sowing, the plants were dried at 90°C for 24 hours, and then the above-ground dry weight of the plants was measured.

[0294] 1st division: control (no application)

[0295] 2-zone: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0296] 3rd division: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in deionized water)

[0297] 4 divisions: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + isobutyl alcohol 50 mass ppm + sorbitan monolaurate 350 mass ppm foliar spray (dissolved in tap water)

[0298] (4) Results

[0299] The aboveground dry weight results are shown in Fig.13 The mean and standard deviation were calculated from the data of 4 replicates excluding the highest and lowest values ​​in each test area with 6 replicates. The graphs in the figures represent the mean ± standard deviation.

[0300] According to Test Example 1 and Example 2, when the same concentration of ascorbic acid is sprayed on the leaves of soybeans, tap water and deionized water have different effects on seed weight and initial growth indicators. It is shown that even in chickpeas, as in soybeans, the effect on initial growth indicators varies 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.

[0301] Example 8: Evaluation of the fruit weight in Lotus japonicus

[0302] (1) Soil preparation

[0303] Mid-term fertilizer-effective soil (Takii Aquatic Cellular Soil Mid-term Fertilizer-effective Type, Takii Seedling Co., Ltd.) and vermiculite granules (Akagi Horticulture Co., Ltd.) were mixed at a volume ratio of 1:1, and the soil was filled in a polyethylene pot (6 cm in diameter, 5.5 cm in height).

[0304] (2) Germination and sowing

[0305] The seeds of Lotus japonicus were collected 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.) containing about 1 / 3 of the seeds, 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 each pot was watered with 100 mL of tap water, 3 seeds that had been treated for germination were sown in each pot, one seed at a time, at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions for each test area was set to 8 (n=8).

[0306] (3) Inoculation of Rhizobia

[0307] 1.5% agar (Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH6.8)) to prepare a solid medium, and Mesorhizobium loti MAFF303099 (MLGUS) strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0308] (3) Cultivation conditions

[0309] The cultivation was carried out indoors under the conditions of 16 hours of daylight, 25°C, LED light source (OHGETS Co., Ltd.; model: VGL-1200W), and light intensity of 400-440 μmol / m 2 / s. After 10 days of sowing, thinning was performed at one plant per pot. After the water in the trough under the pot was empty, watering was performed by adding tap water to the trough to cover the lower part of the pot by about 5 cm. After 3 weeks of sowing, 1,000-fold dilution of high-concentration flowering promotion (HYPONeX JAPAN Co., Ltd.) was added to the water used for watering once a week.

[0310] (4) Foliar spraying

[0311] The spray solution shown in Table 7 was prepared, and 6 weeks after sowing, 6.7 mL was sprayed once per plant in test plots 2 to 6, and 0.125 mL was sprayed once per plant in test plots 7 to 9. The spray solution shown in Table 13 was prepared, and 6.7 mL was sprayed per plant using a mist blower on the 14th day after sowing. Foliar spraying was performed only once, and 7 test plots were studied.

[0312] Regarding the preparation of the spray liquid, the BHT of component (B) was preliminarily dissolved in component (C), and then component (C) and component (D) were mixed to prepare the second dose. Next, component (A) was mixed with other components to prepare the first dose. After the second dose was dissolved in the water to be used, the first dose was further dissolved to prepare the spray liquid. The mature fruits 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.

[0313] Table 7

[0314]

[0315] (5) Results

[0316] 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.

[0317] According to Test Example 1 and Example 2, it was shown that in soybeans, when the same concentration of ascorbic acid was sprayed on the leaves, the effects on seed weight given by tap water and deionized water were different. Even in Lotus japonicus, it was shown that, as with soybeans, the effect on seed weight was different depending on the quality of the water in which ascorbic acid was 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 also increased when tap water was used. According to Test Area 5, when fertilizer components were added to the composition, an increase in yield could also be obtained. In addition, as shown in Test Area 6, a sufficient application effect could also be obtained when the concentration of the composition was halved.

[0318] According to Example 5, in soybean, when a high concentration of the composition is applied, the activity of nodules in the early growth phase is improved. In Lotus japonicus, it was shown that, similar to soybean, by adding component (B), component (C) and component (D) to ascorbic acid, a yield-increasing effect can be obtained when a high concentration and a small amount of water are sprayed using tap water.

[0319] Example 9 Evaluation of the weight of broad beans

[0320] (1) Soil preparation and cultivation conditions

[0321] After filling 2L of potting stones into a planter (width 65.3cm, depth 24.5cm, height 18.5cm), 9L of medium-term fertilizer-effective soil (Takii water-containing cell soil medium-term fertilizer-effective type, Takii Seedling Co., Ltd.) was filled. After 2L of tap water was supplied to each planter, 3 broad bean seedlings (variety: Rende Ichisun, Takii Seedling Co., Ltd.) purchased from a family daily necessities center were planted in each planter. In addition, the number of repetitions for each test area was set to 3 (n=3). Cultivation was carried out outdoors.

[0322] (2) Foliar spraying

[0323] The spray solution was prepared and 6.7 mL was sprayed per plant using a mist blower on the 51st day after seedling planting. Foliar spraying was carried out only once, and four test plots were studied.

[0324] 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 use water, the spray liquid was prepared by further dissolving the first agent. The reagents were the same as those in Example 1. The test areas 1 to 4 evaluated were as follows.

[0325] The seeds were harvested one by one from the mature seeds, and the cultivation was terminated 96 days after planting. The number of beans and the weight of seeds of each plant were measured.

[0326] 1st division: control (no application)

[0327] 2nd division: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in deionized water)

[0328] 3-zone: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0329] 4 divisions: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + isobutyl alcohol 50 mass ppm + sorbitan monolaurate 350 mass ppm foliar spray (dissolved in tap water)

[0330] (3) Results

[0331] The results of the measurement 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 weight of the seeds increased by 32.9%, and the yield-increasing effect was also obtained for broad beans. 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.

[0332] Example 10 Storage stability test of the composition having plant growth promoting and nodule promoting effects and yield increasing effects

[0333] (1) Preparation of one-dose or two-dose composition

[0334] Using ascorbic acid 20g as component (A), BHT13.3mg as component (B), isobutyl alcohol 0.67mL as component (C), and sorbitan monolaurate 4.7mL as component (D), a 1-dose or 2-dose composition is prepared by the following method. After dissolving component (B) in component (C), component (A) is further added to the component mixed with component (D) to prepare a 1-dose composition. The preparation of the 2-dose composition is carried out by the following method. After dissolving component (B) in component (C) in advance, component (D) is further mixed to prepare a second dose. Then, only component (A) is set as the first dose. The reagents are the same as those in Example 1.

[0335] (2) Storage stability test

[0336] The prepared one-dose or two-dose composition was divided into two transparent glass bottles in half. The bottles were stored in a refrigerator set at 4°C or a storage room set at 50°C for 1 week. After 1 week, the appearance was evaluated and the aqueous solution solubility of the stored preparations diluted 500 times with water was evaluated.

[0337] (3) Results of the evaluation of the appearance and solubility of the preparation

[0338] The results of the evaluation of the appearance of the preparations are shown in Fig.18 The left photo shows the first and second doses of the 2-dose composition after storage at 50°C, the center photo shows the first and second doses of the 2-dose composition after storage at 4°C, and the right photo shows the 1-dose composition after storage at 50°C and the 1-dose composition after storage at 4°C.

[0339] according to Fig.18 The results showed that the 2-dose type composition had no difference in appearance from the sample stored at 4°C and the sample stored at 50°C. On the other hand, the 1-dose type composition showed a difference in appearance color depending on the storage temperature. It was also shown that a uniform spray liquid suitable for spraying as a plant growth promoter could be obtained even with either the 1-dose type or the 2-dose type. The results of the study showed that the 1-dose type had a large degree of discoloration during storage, and the 2-dose type was preferred from the perspective of appearance.

[0340] Example 11 Study on the Mixing Ratio of Organic Solvents

[0341] The preparation and sowing of soil, inoculation of rhizobia, the method for recovering the exudate for measuring the amount of ureide nitrogen which is an indicator of nodule activity, the experimental operations before quantifying the exudate, the apparatus and measurement method used in quantifying the amount of ureide nitrogen, the standards used and the items to be quantified were the same as in Example 2, and soybean cultivation was carried out for 21 days.

[0342] (1) Cultivation conditions

[0343] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m 2 / s) 16 hours in the light period / 8 hours in the dark period, the temperature was set to 30°C in the light period / 25°C in the dark period, and the humidity was set to 50%. After 7 days of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to overflow the lower part of the pot by about 5 cm.

[0344] (2) Foliar spraying

[0345] The spray solution shown in Table 8 was prepared and sprayed once with 6.7 mL per plant in test plots 1 to 5 and once with 0.125 mL per plant in test plots 6 to 8 on the 14th day after sowing. Foliar spraying was performed only once, and 7 test plots were studied (Table 8). The reagents used were the same as those in Example 5.

[0346] Table 8

[0347]

[0348] (3) Results

[0349] The results of the nodulation activity assay are shown in Fig.19 Graphs in the figures represent mean ± standard deviation.

[0350] Comparison between test plot 2 and test plots 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 than when ascorbic acid dissolved in tap water is sprayed alone. It also showed that the effect can be exerted even when the mixing ratio of component (B) to component (C) is 1:10 to 10000.

[0351] Example 12 Evaluation of initial growth indicators under low concentration of antioxidant

[0352] The preparation of soil, sowing, and inoculation of rhizobia were carried out in the same manner as in Example 2, and soybean cultivation was carried out for 21 days.

[0353] (1) Cultivation conditions

[0354] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m 2 / s) 16 hours in the light period / 8 hours in the dark period, the temperature was set to 30°C in the light period / 25°C in the dark period, and the humidity was set to 50%. After 7 days of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to overflow the lower part of the pot by about 5 cm.

[0355] (2) Foliar spraying

[0356] Prepare the spray solution, and spray only 6.7 mL per plant once using a mist blower on the 14th day after sowing. For the preparation of the spray solution, after dissolving BHT in isobutyl alcohol, mix isobutyl alcohol and sorbitan monolaurate, and then dissolve them in water. Thereafter, prepare it by dissolving ascorbic acid in water. 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 then the aboveground dry weight of the plant body was measured.

[0357] 1st division: control (no application)

[0358] 2-zone: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0359] 3 divisions: Ascorbic acid 1,500 mass ppm + BHT 0.01 mass ppm + isobutyl alcohol 50 mass ppm + sorbitan monolaurate 350 mass ppm foliar spray (dissolved in tap water)

[0360] (3) Results

[0361] The aboveground dry weight results are shown in Fig. 20 Graphs in the figures represent mean ± standard deviation.

[0362] Comparison between test plots 2 and 3 showed that even when the concentration of BHT in 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 could be obtained than when ascorbic acid dissolved in tap water was sprayed alone.

[0363] Example 13 Evaluation of initial growth index when using anionic surfactant

[0364] The preparation of soil, sowing, and inoculation of rhizobia were carried out in the same manner as in Example 2, and soybean cultivation was carried out for 21 days.

[0365] (1) Cultivation conditions

[0366] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m 2 / s) 16 hours in the light period / 8 hours in the dark period, the temperature was set to 30°C in the light period / 25°C in the dark period, and the humidity was set to 50%. After 7 days of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to overflow the lower part of the pot by about 5 cm.

[0367] (2) Foliar spraying

[0368] Foliar spraying was performed on the 14th day after sowing. In addition to the reagents described in Example 2, sodium dodecyl sulfate, ammonium dodecyl sulfate, and triethanolamine dodecyl sulfate were used as anionic surfactants. The spray solution was prepared by dissolving the reagents in tap water. Foliar spraying was performed only once, and 5 test areas were studied (Table 9). Prepare the spray solution and use a mist blower to spray 6.7 mL per plant. On the 21st day after sowing, the plant body was dried at 90°C for 24 hours, and then the dry weight of the underground part of the plant body was measured.

[0369] Table 9

[0370]

[0371] (3) Results

[0372] The results of underground dry weight determination are shown in Fig.21 The graph in the figure represents the mean value ± standard deviation. Comparison between test areas 2 and 3, 4, and 5 shows that even when anionic surfactants such as sodium lauryl sulfate, ammonium lauryl sulfate, and triethanolamine lauryl sulfate are 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.

[0373] Example 14 Evaluation of soybean nodule activity

[0374] (1) Soil preparation and sowing

[0375] The mid-term fertilizer type soil (Takii water-containing cell soil mid-term fertilizer type, Takii Seedling Co., Ltd.) and vermiculite fine particles (Akagi Horticulture Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a polyethylene pot (diameter 10.5 cm, height 9 cm). Soybean seeds used "エンレイ" (purchased from Nikko Seedling Co., Ltd.). After 250 mL of tap water was given to each pot, 2 seeds were sown in each pot, and they were sown one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions in each test area was set to 6 (n = 6).

[0376] (2) Inoculation of Rhizobia

[0377] 1.5% agar (Fujifilm Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH 6.8)) to prepare a solid medium, and the soybean rhizobium (Bradyrhizobium japonicum) NBRC14783T strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0378] (3) Cultivation conditions

[0379] Cultivation from sowing to thinning was carried out using an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (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 of sowing, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to cover the lower part of the pot by about 5 cm.

[0380] (4) Foliar spraying

[0381] The spray solution was prepared and 6.7 mL was sprayed per plant using a mist blower on the 14th day after sowing. Foliar spraying was performed only once, and three test plots were studied. The test plots 1 to 3 evaluated are as follows.

[0382] <Spray liquid>

[0383] 1st division: control (no application)

[0384] · 2nd division: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0385] · 3-partition: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + diethylene glycol 200 mass ppm + sorbitan monolaurate 350 mass ppm for foliar spraying (dissolved in tap water)

[0386] (5) Method for recovering exudate for measuring the amount of ureide nitrogen as an indicator of nodule activity

[0387] After 21 days of cultivation, the soybean stems were cut at the cotyledonary node using flower and tree scissors, and covered with a 1.5 mL microtube (Eppendorf Co., Ltd.) plugged with a cotton ball #10 (Iwatsuki Co., Ltd.), and the exudate flowing out from the cut surface of the stem was collected for 2 hours. The cotton ball after the exudate was collected was stored in a freezer at -80°C. In addition, the amount of exudate was calculated by comparing the weight of the cotton ball before and after the exudate was collected.

[0388] (6) Extraction of exudate components recovered from cotton balls

[0389] The exudate components contained in the cotton ball were eluted with ultrapure water and filtered using a microbial spin chromatography column (Bio-Rad Laboratories Co., Ltd.). The filtrate remaining in the column was centrifuged and recovered by running a centrifuge (CR15RN, Hitachi, Ltd.) at 10,000 rpm for 1 minute. The filtrate diluted 3,000 times was quantified.

[0390] (7) Apparatus and measurement method used for quantification of ureide nitrogen content

[0391] The HPLC device and mass spectrometer used Agilent 1260 Infinity LC system (Agilent Technologies Co., Ltd.) and AB SCIEX TripleQuad 4500 system (AB Sciex Co., Ltd.), respectively. The chromatographic column used Scherzo SS-C18 (100 mm × 2 mm, 3 μm) (Imtakt Co., Ltd.), and the chamber temperature was set to 40°C. 5 μL of appropriately diluted sample was injected, the flow rate was 0.5 mL / min, and the eluent used 0.1% formic acid aqueous solution (eluent A) and 50 mM ammonium acetate / methanol solution (eluent B). After balancing with eluent A: eluent B = 95:5, the sample was supplied, and after 5 minutes, the ureide nitrogen was eluted by a linear gradient of eluent A: eluent B = 80:20.

[0392] (8) Standards used and quantification

[0393] The amount of ureide nitrogen in the exudate was analyzed by LC-MS. Allantoic acid (Toronto Research Chemicals Inc.) was used as a standard. The standard was analyzed by LC-MS, and a calibration curve was prepared in the range of 10-1000 ppb.

[0394] For each sample, allantoic acid was identified based on the retention time, precise mass, and MS / MS spectrum of each reagent. In addition, allantoic acid in the sample was quantified based on the calibration curve to determine the total amount per plant.

[0395] (9) Results

[0396] The results of the nodulation activity assay are shown in Fig.30 The graphs in each figure represent the mean ± standard deviation.

[0397] Comparison between test plots 2 and 3 showed that the addition of component (B), component (C) and component (D) to ascorbic acid resulted in a higher nodule activity enhancement effect than when ascorbic acid dissolved in tap water was sprayed alone.

[0398] Example 15 Effect on soybean growth

[0399] (1) Soil preparation and sowing

[0400] The mid-term fertilizer type soil (Takii water-containing cell soil mid-term fertilizer type, Takii Seedling Co., Ltd.) and vermiculite fine particles (Akagi Horticulture Co., Ltd.) were mixed in a volume ratio of 1:1, and the soil was filled into a polyethylene pot (diameter 10.5 cm, height 9 cm). Soybean seeds used "エンレイ" (purchased from Nikko Seedling Co., Ltd.). After 250 mL of tap water was given to each pot, 2 seeds were sown in each pot, and they were sown one by one at a depth of about 1 to 2 cm from the soil surface. In addition, the number of repetitions in each test area was set to 6 (n = 6).

[0401] (2) Inoculation of Rhizobia

[0402] 1.5% agar (Fujifilm Wako Pure Chemical Industries, Ltd.) was added to Yeast-Mannitol (YM) medium (K2HPO4 0.5g, MgSO4·7H2O 0.2g, NaCl 0.1g, Yeast Extract 0.4g, Mannitol 10g, distilled water 1L (pH 6.8)) to prepare a solid medium, and the soybean rhizobium (Bradyrhizobium japonicum) NBRC14783T strain was grown on the solid medium. An inoculation loop of the grown rhizobia was taken and inoculated on 50mL of YM liquid medium in a 500mL Sakaguchi flask, and cultured with shaking at 30°C for about 36 hours. A rhizobium culture solution with a turbidity OD600 value of about 0.3 was prepared. After sowing, 1mL of the rhizobium culture solution was dropped on the seeds using a micropipette for inoculation.

[0403] (3) Cultivation conditions

[0404] The cultivation was carried out in an artificial weather machine (LPH-411SP, Nippon Ikagi Seisakusho Co., Ltd.), and the light conditions were set to the bright period (light intensity 440-480 μmol / m2 / s) 16 hours / dark period 8 hours, temperature set to 30°C in light period / 25°C in dark period, humidity set to 50%. After germination, thinning was performed with one plant per pot. After the water in the trough under the pot was gone, tap water was added to the trough to cover the lower part of the pot by about 5 cm.

[0405] (4) Foliar spraying

[0406] The spray solution was prepared and 6.7 mL was sprayed per plant using a mist blower on the 7th day after sowing. Foliar spraying was performed only once, and five test plots were studied. The test plots 1 to 5 evaluated were as follows.

[0407] <Spray liquid>

[0408] 1st division: control (no application)

[0409] · 2nd division: 1,500 ppm by mass of ascorbic acid for foliar spraying (dissolved in tap water)

[0410] · 3rd division: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + glycerin 50 mass ppm (dissolved in tap water)

[0411] 4 divisions: Ascorbic acid 1,500 mass ppm + BHT 1 mass ppm + Propylene glycol 50 mass ppm (dissolved in tap water)

[0412] 5th division: Sodium ascorbate 1,500 mass ppm + tocopherol 10 mass ppm + glycerol 50 mass ppm (dissolved in tap water)

[0413] (5) Results

[0414] On the 9th day after sowing, the plants were dried at 100°C for 7 days, and then the aboveground dry weight of the plants was measured. The results of the aboveground dry weight measurement are shown in Fig.31 . The graphs in each figure represent the mean value ± standard deviation. Comparison of test plots 1 to 5 showed that no growth-promoting effect was observed when ascorbic acid was dissolved in tap water and sprayed on the leaves, but by adding component (B) and component (C) to ascorbic acid or sodium ascorbate, a growth-promoting effect was obtained when tap water was used as the dissolving water. Compared with the control, the aboveground dry weight increased by an average of about 3.7% in test plot 3, about 9.1% in test plot 4, and about 2.1% in test plot 5.

Claims

1. A composition, which is a growth promoter composition for leguminous plants contained in beans, wherein: The composition is a composition comprising a first agent containing (A) ascorbic acid or a salt thereof and a second agent containing (B) an antioxidant and (C) an organic solvent. (B) the antioxidant is at least one selected from 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, (A) the mass ratio of ascorbic acid or a salt thereof is 10 to 3,000,000 when the component (B) is 1, or (C) the mass ratio of the organic solvent is 4 to 100,000 when the component (B) is 1, The first and second doses are respectively contained in one of the two-headed bottles formed by connecting the first and second bottles via a connecting member.

2. The composition according to claim 1, wherein The first bottle body includes: a bottle body having an external threaded portion at the lower portion; a narrow-mouth bottle head portion; and a cap body, and the second bottle body includes a bottle body and a wide-mouth bottle head portion.

3. The composition according to claim 1 or 2, wherein The first dose is contained in the second bottle, and the second dose is contained in the first bottle.

4. A double-headed bottle, which is a bottle for containing the composition according to claim 1, wherein: The first bottle body comprises: a bottle body having an external threaded portion at the bottom; a narrow-mouthed bottle head; and a cap body. The second bottle body comprises a bottle main body and a wide-mouth bottle head. The wide-mouth bottle head of the second bottle body is connected to the lower part of the first bottle body via a connecting member.

5. The bottle according to claim 4, wherein: The connecting member comprises: a cover plate; a first ring wall extending vertically upward along the outer peripheral edge of the cover plate; and a second ring wall formed by the first ring wall extending horizontally outward and then folding vertically downward. The external threaded portion of the first bottle body arranged at the lower part of the bottle main body having an external threaded portion at the lower part is screwed into the internal threaded portion of the connecting component arranged on the inner wall of the first ring wall, and the external threaded portion of the second bottle body arranged at the wide-mouth bottle head is screwed into the internal threaded portion of the connecting component arranged on the inner wall of the second ring wall, thereby connecting the first bottle body and the second bottle body.

6. The bottle according to claim 4, wherein: The connecting member comprises a cover plate and a first annular wall and a second annular wall respectively protruding toward the outer peripheral edge of the cover plate in a vertical direction. The external threaded portion of the first bottle body arranged at the lower part of the bottle main body having an external threaded portion at the lower part is screwed into the internal threaded portion of the connecting component arranged on the inner wall of the first ring wall, and the external threaded portion of the second bottle body arranged at the wide-mouth bottle head is screwed into the internal threaded portion of the connecting component arranged on the inner wall of the second ring wall, thereby connecting the first bottle body and the second bottle body.

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