WO2018159393A1 - マメ科植物生育促進剤 - Google Patents
マメ科植物生育促進剤 Download PDFInfo
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- WO2018159393A1 WO2018159393A1 PCT/JP2018/006081 JP2018006081W WO2018159393A1 WO 2018159393 A1 WO2018159393 A1 WO 2018159393A1 JP 2018006081 W JP2018006081 W JP 2018006081W WO 2018159393 A1 WO2018159393 A1 WO 2018159393A1
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- plant
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- soyasapogenol
- glycoside
- soybean
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/20—Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/10—Fertilisers containing plant vitamins or hormones
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/256—Polyterpene radicals
Definitions
- the present invention relates to an ingredient that promotes the growth of legumes.
- Non-Patent Document 1 Increasing the root weight, that is, the underground weight, not only directly increases the nutrient supply capacity (source capacity), but also contributes to the prevention of aboveground lodging. It is expected as a way to increase.
- nodules are known as an underground source (nitrogen source) characteristic of legumes.
- Non-patent Document 2 In soybean, it has been shown that an increase in the number of nodules and nodule weight leads to an increase in yield (Non-patent Document 2).
- Technology that promotes nodulation is expected to achieve increased yields while suppressing the use of chemical fertilizers, and is therefore desirable from the viewpoint of building a sustainable agricultural production system.
- Saponins are a type of glycoside contained in various plants and have been used as surfactants, emulsifiers and the like. The structures of saponins are roughly classified into triterpenoid saponins and steroid saponins, but the types are extremely diverse.
- saponins The diverse structures of saponins reflect their diverse physiological activities. In other words, saponins exert various physiological actions on animals and plants, but their actions seem to differ depending on the structure. For example, it has been reported that Quillaja saponin (Patent Document 1) and saponin extracted from loofah, ginseng, cucumber, melon or amacha-kul (Patent Document 2) promotes plant growth or increases yield. Moreover, it is described that saponin derived from tea seeds promotes infection of VA mycorrhizal fungi (VesicularesArbuscular Mycorrhizae), which promotes plant growth (Patent Document 3).
- Soyasaponins which are a kind of saponins, are a kind of oleanane-type triterpenoid saponins and are characteristic metabolites contained in legumes. However, there are many types of soyasaponins, and their characteristics are also very different from each other.
- glycosides of Soyasapogenol B glycosides (so-called Soyasaponins of Group B group) in which a sugar is bonded to the C-3 position hydroxy group of Soyasapogenol B and the C-22 position is a hydroxy group
- Soyasapogenol B The glycosides (so-called DDMP saponins) in which a sugar is bonded to the C-3 position hydroxy group and maltol is bonded to the C-22 position hydroxy group are greatly different in characteristics.
- Non-patent Document 3 DDMP saponin was involved in removal of active oxygen and promotion of root elongation in germinated soybeans under experimentally controlled conditions, but group B group soyasaponin had an effect on root growth. It has been reported that there was not.
- Non-Patent Document 4 reports that crude saponin derived from mungbean (Yenenari) promoted the germination rate and initial growth of mungbean but did not increase the yield.
- Non-Patent Documents 5 and 6 report reduction of mung bean seedling size by soyasaponin of group B group derived from mung bean and inhibition of wheat seedling growth by alfalfa-derived saponin.
- Non-patent documents 7 to 10 also report the effects of various soyasaponins on plant growth, but it is suggested that the effects differ depending on the plant species and the structure of soyasaponins.
- soyasaponin ⁇ g also referred to as soyasaponin VI or chromosaponin I
- DDMP saponin which is a DDMP saponin
- group B group soyasaponin Bb also referred to as soyasaponin I
- DDMP saponin is chemically unstable, and its production requires a strict extraction process (Non-patent Document 11). Therefore, the practicality of DDMP saponins in the cultivation of plants in actual agriculture is extremely low due to problems in production and stability during application.
- Patent Document 1 JP-A No. 2004-121186 (Patent Document 2) JP-A No. 61-15806 (Patent Document 3) JP-A No. 8-23963 (Non-Patent Document 1) Japanese Society of Breeding and Japanese Crop Society Hokkaido discourse bulletin, 1992, (31): 64 (Non-patent document 2) Crop research report, 2007, (8): 49-108 (Non-patent document 3) Seed science and biotechnology, Seed Physiology and Biochemistry Study Group (edited), Academic Publishing Center, 2009, pp.106-112 (Non-Patent Document 4) Botanical Bulletin of Academia Sinica, 1995, 36 (1): 9-18 (Non-Patent Document 5) Advances in Plant Glycosides, Chemistry and Biology, Volume 6, Elsevier Science, 1999, pp.105-130 (Non-patent document 6) Plant and Soil, 1987, 98 (1): 67-80 (Non-patent document 7) Physiol Plantarum, 1995
- Non-Patent Document 9 Isoprenoid Synthesis in Plants and Microorganisms: New Concepts and Experimental Approaches, Springer, 2013, pp.405-424 (Non-Patent Document 10) Phytochem Rev, 2013, (12): 877-893 (Non-Patent Document 11) Journal of Soy Protein Research Association, 1994, (15): 36-40
- the present invention provides a glycoside of Soyasapogenol B, wherein the C-22 position of Soyasapogenol B is a hydroxy group, and a sugar is bonded to the C-3 position hydroxy group of Soyasapogenol B.
- the present invention provides a leguminous plant growth promoter comprising a glycoside as an active ingredient.
- the present invention relates to a glycoside of Soyasapogenol B, wherein the C-22 position of Soyasapogenol B is a hydroxy group, and a sugar is bonded to the C-3 position hydroxy group of Soyasapogenol B.
- the present invention provides a method for promoting the growth of legumes using the existing glycoside as an active ingredient.
- Rhizobium Rhizobium material only, Rhizobium + Gen: Rhizobium material + genistein application group, Rhizobium + SSB: Rhizobium material + soyasaponin Bb application group.
- Non-application area soil only, rhizobia: only rhizobial material, rhizobia + S50: rhizobial material + soybean saponin preparation 50 ppm, rhizobia + S100: rhizobia material + soybean saponin preparation 100 ppm, rhizobia + S500: rhizobia material + soybean Saponin preparation 500 ppm.
- the labels on the horizontal axis are the same as in FIG.
- Non-application section saponin preparation non-application section
- preparation saponin preparation application section.
- Non-application area saponin preparation non-application area
- saponin saponin preparation application area
- catechin catechin preparation application area
- combination area saponin-catechin combination area.
- Non-application area saponin preparation non-application area
- saponin saponin preparation application area
- iron phosphate iron phosphate (III) application area
- combination area saponin-iron phosphate (III) combination area.
- the labels on the horizontal axis are the same as in FIG.
- Non-application area nutrient solution only, 10 ppm: saponin preparation 10 ppm application area, 100 ppm: saponin preparation 100 ppm application area.
- Non-application zone soil only, 10 ppm: saponin formulation 10 ppm application zone, 100 ppm: saponin formulation 100 ppm application zone.
- the present invention provides ingredients that promote the growth of legumes.
- the leguminous plant growth promoter of the present invention promotes the growth of legumes and improves the productivity of legumes, which are important crops.
- a group B group of soyasaponins that is, glycosides of soyasapogenol B, wherein the C-22 position of the soyasapogenol B is a hydroxy group, and a sugar is present in the C-3 position hydroxy group of the soyasapogenol B).
- Glycosides are used as active ingredients to promote the growth of legumes.
- “Growth promotion” of legumes means, for example, an increase in the weight of the above-ground and underground parts, an increase in the number of lateral buds, an increase in yield, and the promotion of nodulation (for example, Any one or more selected from the group consisting of an increase or an increase in nodule weight).
- “yield” is selected from the group consisting of the number of flowers, the number of pods, the weight of the seeds, the weight of seeds, and the increase of the total weight of seeds per unit area or unit area (so-called seed yield). Any one or more.
- the “growth promotion” of legumes according to the present invention is any one or more selected from the group consisting of an increase in underground weight, an increase in the number of lateral buds, an increase in yield, and promotion of nodulation. More preferably, it means an increase in yield, especially an increase in the total weight of seeds per seed or unit area (so-called seed yield).
- the “above-ground part” and “underground part” of legumes are respectively higher than the upper surface of a cultivation base (for example, soil, water, nutrient solution, medium, etc.) in the legume plant body.
- a cultivation base for example, soil, water, nutrient solution, medium, etc.
- leguminous plant examples include, for example, a plant belonging to the genus Soybean (Glycine), a plant belonging to the genus Bean, a chick genus plant, a pea genus plant, a genus Lens, a genus Cajanus. Plants, Vicia genus plants, Arachis genus plants, Medicago genus plants, Nepitonia genus plants, Trigonella genus plants, Psophocarpus genus plants, etc.
- Glycine genus Soybean
- Preferred examples include soybean plant, kidney bean plant, chickpea plant, pea plant, lentil plant, bean plant, broad bean plant, and peanut plant, more preferred examples Is to, soybean plant of the genus Phaseolus plants, chickpea plants of the genus, include pea plants of the genus, as more preferred examples include soybean plants of the genus.
- soybean genus plants include soybean (Glycine max), examples of common bean plants, common bean (Phaseolus vulgaris), examples of chickpea plants, chickpea (Cicer arietinum), examples of pea plant
- a pea Pieris sativum
- a lentil genus plant as for an example of a lentil (Lens culinaris)
- a genus of bean genus (Cajanus cajan)
- a broad bean genus plant Is a broad bean (Vicia faba)
- an example of a peanut plant is Arachis hypogaea
- an example of a coconut genus plant is an alfalfa (Medicago ⁇ sativa)
- Neptunia oleracea is an example of the plant belonging to the genus Fenugreek, Trigonella foenum-g raecum)
- winged genus plants include winged bean
- leguminous plants to be promoted by the present invention include soybean plant, kidney bean plant, chickpea plant, pea plant, lentil plant, bean plant, broad bean plant, and peanut plant. Is at least one selected from the group consisting of In a more preferred embodiment, the leguminous plant to be promoted by the present invention is at least one selected from the group consisting of soybean, kidney bean, chickpea, pea, lentil, bean, broad bean and groundnut. More preferably, the leguminous plant to be promoted for growth according to the present invention is at least one selected from the group consisting of soybean, kidney bean and chickpea, and more preferably soybean.
- the soyasaponin of group B group means that the C-22 position of soyasapogenol B represented by the following formula (I) is a hydroxy group, and a sugar is bonded to the C-3 position hydroxy group of soyasapogenol B. It is a glycoside of soyasapogenol B.
- a glycoside in which 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one is bonded to the C-22 hydroxy group of soyasapogenol B is The soyasaponins in Group B are not classified.
- R represents a sugar residue or a sugar chain.
- examples of the sugar residue represented by R include glucuronic acid, galactose, glucose, rhamnose, arabinose, and the like.
- sugar chain represented by R include rhamnose (1 ⁇ 2).
- Examples of the compound represented by the formula (I) include Soyasaponin Bb (also referred to as Soyasaponin I), Soyasaponin Bc (also referred to as Soyasaponin II) and Soyasaponin Ba (also referred to as Soyasaponin V), and more preferably Soyasaponin Bb. Is mentioned.
- the structures of soyasaponin Bb, soyasaponin Bc and soyasaponin Ba are represented by the following formulas (II), (III) and (IV), respectively.
- the active ingredient for promoting the growth of legumes used in the present invention is a hydroxy group at the C-22 position of soyasapogenol B, and a sugar is bonded to the C-3 hydroxy group of soyasapogenol B.
- Glycoside of Soyasapogenol B preferably one or more selected from the group consisting of compounds represented by the above formula (I), more preferably selected from the group consisting of Soyasaponin Bb, Soyasaponin Bc and Soyasaponin Ba Or more, and more preferably Soyasaponin Bb.
- the group B group soyasaponins used in the present invention can be prepared by extraction or purification from legumes such as soybean, or secretory production from legumes such as soybean.
- Group B is obtained by pulverizing legume seeds such as dried soybeans, extracting with a solvent such as ethanol, and further purifying through a column or resin as necessary.
- a group of soyasaponins can be isolated.
- a group B group soyasaponin secreted into a hydroponic culture solution of a leguminous plant such as soybean can be used.
- Group B group soyasaponins eg, available from ChromaDex, Inc.
- a commercially available soy saponin preparation rich in soyasaponin of Group B group eg Wako Pure Chemical Industries, Ltd., Access One Co., Ltd., J-Oil Mills, Fuji Oil Co., Ltd.
- Tokiwa Phytochemical Laboratory available from FAP Japan Co., Ltd.
- the present invention provides a leguminous plant growth promoter comprising group B group soyasaponin as an active ingredient.
- the present invention provides the use of Group B group Soyasaponins for the manufacture of legume growth promoters.
- the present invention provides the use of Group B group Soyasaponins to promote legume growth.
- the present invention provides group B group soyasaponins for use in promoting legume growth.
- the present invention provides a method for promoting the growth of legumes using Group B group soyasaponins.
- the growth promotion preferably refers to any one or more selected from the group consisting of an increase in the weight of the underground part, an increase in the number of side buds, an increase in yield, and promotion of nodulation, and more preferably An increase in yield, especially an increase in the total seed weight per line or unit area.
- group B group soyasaponin may be used alone or in the form of a composition containing group B group soyasaponin as an active ingredient. Therefore, the form of the leguminous plant growth promoter provided by the present invention is not particularly limited.
- the group B group soyasaponin alone may be used, but the group B group soyasaponin is included as an active ingredient. It may be a composition (for example, various agricultural or horticultural materials).
- the growth promoter of the present invention can have any form such as a block, powder, granule, liquid, or gel.
- the group B group soyasaponin When using a group B group soyasaponin as a simple substance, the group B group soyasaponin is added to a cultivation base for cultivating legumes such as soil, a medium, a solution for hydroponics, or the like. What is necessary is just to prepare the water and additive containing soyasaponin of the group B group, and to add this water and additive to the cultivation base material for this legume. Or the water and additive containing the soyasaponin of the group B group may be prepared, and the water and additive may be given to the legume seeds and plants.
- composition containing group B group soyasaponin as an active ingredient examples include, for example, a cultivation substrate containing group B group soyasaponin as an active ingredient (eg, agricultural or horticultural soil, soil, culture medium, nutrient solution) Cultivation solutions, water, etc.), fertilizers, water for watering, microbial materials such as rhizobial materials, soil improvers, pesticides, sowing materials, plant supplements (eg, activators, nutrients, etc.), etc.
- a cultivation substrate containing group B group soyasaponin as an active ingredient eg, agricultural or horticultural soil, soil, culture medium, nutrient solution) Cultivation solutions, water, etc.
- fertilizers water for watering
- microbial materials such as rhizobial materials, soil improvers, pesticides, sowing materials
- plant supplements eg, activators, nutrients, etc.
- a fertilizer, microbial material, soil improver, sowing material, and plant supplement containing soyasaponin of Group B group as an active ingredient is preferable because it contributes to the improvement of the soil on which legumes are grown.
- the fertilizer, microbial material, soil improver, sowing material, and plant supplement may be solid or liquid.
- the fertilizer, microbial material, soil conditioner, sowing material, and plant supplement are solid, they can be in the form of blocks, powders, granules, etc., but are preferably powders or granules.
- the fertilizer, microbial material, soil improver, sowing material, and plant supplement include fertilizer, microbial material, and soil improver that are usually used for cultivation of legumes, in addition to containing group B group soyasaponin as an active ingredient Ingredients for sowing, plant supplements.
- the microbial material is preferably a rhizobial material.
- the species of rhizobia contained in the rhizobial material can be selected according to the leguminous plant to be cultivated. Table 1 below illustrates examples of rhizobia that can be included in the rhizobial preparation used in the present invention and leguminous plants that are targets thereof.
- blended with microbial material are also called microbial cells.
- any one of the rhizobia species listed above may be used alone for each target leguminous plant, or any two or more may be used in combination.
- -For soybean plants one or more selected from the group consisting of the genus Bradyrhizobium and Ensifer (Sinorhizobium) is preferred, and the Bradyrhizobium genus is more preferred.
- the Bradyrhizobium genus is preferably at least one selected from the group consisting of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens and Bradyrhizobium elkanii, more preferably at least one selected from the group consisting of Bradyrhizobium japonicum and Bradyrhizobium diazoefficiens.
- Ensifer (Sinorhizobium) is preferred; -Rhizobium spp. are preferred for the use in the kidney bean plant, the pea plant, the broad bean plant, the lentil plant, and the bean plant.
- the Rhizobium genus is preferably one or more selected from the group consisting of Rhizobium leguminosarum, Rhizobium gallicum and Rhizobium giardinii, more preferably Rhizobium leguminosarum; -For chickpea plants, the genus Mesorhizobium is preferred, and the genus Mesorhizobium is preferably at least one selected from the group consisting of Mesorhizobium ciceri and Mesorhizobium mediterraneum, more preferably Mesorhizobium ciceri; -For peanut plants, Bradyrhizobium is preferred.
- the preferred species of the genus Bradyrhizobium is the same as in the case of soybean plants.
- a material of a microbial species other than rhizobia may be used, or a material combining a microbial species other than rhizobia and a rhizobia may be used.
- microbial species other than rhizobia include plant growth promoting rhizosphere bacteria and plant growth promoting fungi.
- Examples of the plant growth promoting rhizobacteria include, for example, Bacillus, Pseudomonas, Azospirillum, Burkholderia, Enterobacter, Talaromyces, Arthrobacter, Agrobacterium, Corynebacterium, Erwinia Fungi, Psychrobacter genus, Serratia genus and Rhodococcus genus; examples of the plant growth promoting fungi include Penicillium, Trichoderma, Fusarium, Phoma, Glomus, Acaulospora Entrophospora, Gigaspora, Scutellospora and Aspergillus.
- Cultivation base materials, soy saponins of Group B group as active ingredients, fertilizers, microbial materials such as rhizobial materials, soil improvers, pesticides, sowing materials, plant supplements are used for normal cultivation base materials (for example, for agriculture) Or horticultural soil, soil, culture medium, solution for hydroponics, water, etc., fertilizer, microbial materials such as rhizobial materials, soil improvers, agricultural chemicals, sowing materials, plant supplements (eg, activators, It may be prepared by adding a group B group soyasaponin to a nutrient or the like.
- the concentration of the group B group soyasaponin in the composition containing the group B group soyasaponin as an active ingredient is, for example, when the composition is a fertilizer, a microbial material, a soil conditioner, a sowing material, a plant supplement, etc.
- the composition preferably 0.0005% by mass or more, more preferably 0.005% by mass or more, and preferably 80% by mass or less, more preferably 50% by mass or less, and further preferably 5% by mass. Or less, more preferably 0.5% by mass or less, or preferably 0.0005 to 80% by mass, more preferably 0.0005 to 50% by mass, 0.0005 to 5% in the total amount of the composition.
- the concentration of group B group soyasaponin in the composition is preferably 0.01 ppm by mass or more, more preferably 0.1 ppm by mass or more, in the total amount of the composition. And preferably 100 ppm by mass or less, more preferably 10 ppm by mass or less, further preferably 5 ppm by mass or less, more preferably 2 ppm by mass or less, or preferably in the total amount of the composition.
- 0.01-100 mass ppm more preferably 0.01-10 mass ppm, 0.01-5 mass ppm, 0.01-2 mass ppm, 0.1-100 mass ppm, 0.1-10 mass ppm 0.1 to 5 ppm by mass or 0.1 to 2 ppm by mass.
- the amount of group S group soyasaponin for promoting the growth of legumes according to the present invention is preferably 0.01 to 100 ppm by mass as the concentration in the cultivation base for cultivating legumes.
- the amount of group B group soyasaponin used per 1 liter of cultivation base is preferably 0.01 to 100 mg, more preferably 0.01 to 50 mg, 0.01 to 20 mg, 0.01 to 13 mg, 0 .1 to 10 mg, 0.1 to 5 mg, or 0.1 to 2 mg may be used.
- Group B group soyasaponins may be added to the soil in amounts of .01 to 1 kg, 0.01 to 0.5 kg, or 0.01 to 0.2 kg. That is, in the case of a composition containing soyasaponin of group B group as an active ingredient, such as fertilizer, microbial material, soil conditioner, sowing material, plant supplement, etc., the amount of the composition used in the composition It depends on the concentration of soyasaponin in the group B included.
- the amount of the composition used per 10 ares of land is preferably 20 to 200,000 kg, more preferably 20 to 100,000 kg, 20 to 40,000 kg, 20 to 26,000 kg, 200 to 20,000 kg, 200 to 10,000 kg, or 200 to 4,000 kg.
- leguminous plant cultivation techniques include hydroponics, spray cultivation, sand cultivation, gravel cultivation, etc., soil cultivation, soil culture cultivation, etc.
- Soil cultivation is preferred.
- soil cultivation it is preferable to use agricultural or horticultural soil or soil.
- the soil or soil used for soil cultivation is preferably subjected to soil improvement such as agglomeration treatment.
- a soil improving agent for agglomeration treatment of soil or soil a soil improving agent containing alkali-treated lignin as an active ingredient described in JP-A-2017-190448 is preferable.
- the cultivation of legumes in the present invention is preferably indoor cultivation from the viewpoint of environmental stability, but outdoor cultivation is more preferred from the viewpoint of securing the yield.
- the method for promoting the growth of legumes according to the present invention is a group B group soyasaponin (in the form of a simple substance or a composition containing the above-mentioned group B group soyasaponin as an active ingredient). ) As an active ingredient.
- the leguminous plant is cultivated together with the group B group soyasaponin.
- the group B group soyasaponin is preferably added to a cultivation substrate (for example, soil, soil, culture medium, solution for hydroponics, water, etc.).
- a cultivation substrate for example, soil, soil, culture medium, solution for hydroponics, water, etc.
- a legume plant is cultivated by a normal procedure on a cultivation base material containing the group B group soyasaponin, the effect of promoting the growth of the leguminous plant by the group B group soyasaponin can be obtained.
- fertilizer or rhizobial material normally applied to legumes may be added to the soil as needed, and the group B group soyasaponin may be applied thereto.
- the group B group soyasaponins are used in the form of fertilizers, microbial materials, etc., it is not necessary to add fertilizers, microbial materials, etc. separately, but they may be combined with the addition of other fertilizers, microbial materials, etc. If the leguminous plant is cultivated in a normal procedure in the prepared soil to which the group B group soyasaponin is added, the growth promoting effect by the group B group soyasaponin can be obtained. In the case of hydroponics, the soyasaponin of the group B group may be added to the nutrient solution.
- the group B group soyasaponin is preferably added to the cultivation substrate before sowing, but may be added after sowing, or may be added both before and after sowing.
- the group B group soyasaponins are preferably applied by applying or smearing seeds before sowing (for example, as seed dressing). If seeds to which soyasaponin of group B group is added are cultivated on a cultivation base by a normal procedure, the growth promotion effect of legumes by soyasaponin of group B group can be obtained.
- the addition of the group B group soyasaponin to the cultivation substrate eg, soil, soil, culture medium, hydroponic solution, water, etc.
- the seeds before sowing may be combined with the application or smearing of the group B group soyasaponin.
- the Group B group soyasaponins may be provided by direct application, spraying or application (eg foliar application) to legumes. If the leguminous plant to which the group B group soyasaponin is added is cultivated by a normal procedure, the growth promoting effect of the leguminous plant by the group B group soyasaponin can be obtained.
- the addition of the group B group soyasaponin to the cultivation substrate eg, soil, soil, culture medium, hydroponic solution, water, etc.
- the cultivation substrate eg, soil, soil, culture medium, hydroponic solution, water, etc.
- application or smearing of the group B group soyasaponin to the seed before sowing may be combined with direct application, spraying or application of the group B group soyasaponin to the legume plant.
- the cultivation period of the leguminous plant according to the method of the present invention is preferably 20 days or more from the sowing of the seed, and more preferably until the seed can be harvested.
- the period required until the seed can be harvested depends on the type of legumes and the cultivation environment.
- group B group soyasaponins are other components for promoting leguminous plant growth, such as plant essential nutrients, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleobases, sugars, monohydric alcohols.
- Nonionic surfactant Food additive, microbial extract, plant hormone, nod factor ie lipo-chitooligosaccharide, synthetic lipo-chitooligosaccharide, chitooligosaccharide, chitinous compound, linoleic acid or its derivatives, linolenic acid or It may be used in combination with one or more selected from derivatives thereof, calikines, acyl-homoserine lactone derivatives, betaine compounds, phenol compounds and the like.
- These other components may be contained in a composition containing the group B group of soyasaponins described above, or may be applied separately to legumes. The amount of these other components used may be an amount that does not inhibit the leguminous plant growth promoting effect of group B group soyasaponins.
- Examples of essential nutrients of the plant include nitrogen, phosphorus, potassium, calcium, sulfur, magnesium, boron, chlorine, manganese, iron, zinc, copper, molybdenum, nickel, and the like.
- Examples of the flavonoid include genistin and daidzin.
- Examples of the organic acid include citric acid, oxalic acid, ferulic acid, caffeic acid, malic acid, malonic acid, picidic acid, mugineic acid, dehydroxymugineic acid, hydroxymugineic acid, and acetic acid.
- amino acids examples include serine, proline, leucine, isoleucine, methionine, cysteine, tryptophan, asparagine, glutamine, aspartic acid, glutamic acid, and 5-aminolevulinic acid; Guru Examples include thione, glycopeptide, soybean protein degradation product, plant-derived polypeptide, etc .; examples of the nucleoside include inosine, guanosine, uridine, etc .; examples of the nucleotide include inosinic acid, guanylic acid, uridylic acid, etc.
- nucleobase examples include hypoxanthine, guanine, and uracil
- examples of the sugar include trehalose, sucrose, glucose, and maltose
- examples of the monohydric alcohol include alcohol lauryl.
- nonionic surfactants include polyoxyethylene oleyl ether, polyoxyethylene lauryl ether, and polyoxyethylene.
- examples of the food additive include chitosan; examples of the microorganism extract include yeast extract; and examples of the plant hormone include indole-3-acetic acid and indole.
- Synthetic lipo-chitooligosaccharides include, for example, synthetic LCO compounds described in WO2005 / 063784, and AcNodRM-1 and AcNodRM-3 described in US Pat. No.
- sugar examples include oligo-N-acetylglucosamine; examples of the chitinous compound include chitin and chitosan; examples of the linoleic acid and derivatives thereof include linoleic acid; Examples of the linolenic acid or its derivatives include Examples of the calicin include hydrochloride and hydrobromide; examples of the acyl-homoserine lactone derivative include N-acyl-L-homoserine lactone and N-hexanoyl-L-homoserine lactone.
- betaine compound examples include N, N, N-trimethylglycine, carnitine and the like; Examples include cresol and chlorophenol.
- group B group soyasaponins for promoting legume growth according to the present invention are used in combination with catechins.
- catechins include catechin (C), gallocatechin (GC), catechin gallate (Cg), gallocatechin gallate (GCg), epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECg). And at least one selected from the group consisting of epigallocatechin gallate (EGCg).
- the catechins can be extracted from tea leaves, grapes, cacao beans or processed products thereof produced from Camellia plants (for example, C. sinensis var. Sinensis, C. sinensis var. Assamica, etc.).
- soyasaponin group B group for the leguminous plant growth promotion by the present invention is combined with iron phosphate (III) (FePO 4).
- [2] Preferably essential plant nutrients, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleobases, sugars, monohydric alcohols, nonionic surfactants, food additives, microbial extracts, plant hormones, Nod factors, ie lipo-chitooligosaccharides, synthetic lipo-chitooligosaccharides, chitooligosaccharides, chitinic compounds, linoleic acid or derivatives thereof, linolenic acid or derivatives thereof, calikines, acyl-homoserine lactone derivatives, betaine compounds, and phenolic compounds
- the leguminous plant growth promoter according to [6], wherein the cultivation base material is preferably soil, culture medium, culture medium, hydroponic solution or water.
- Growth promotion is preferably one or more selected from the group consisting of an increase in underground weight, an increase in the number of lateral buds, promotion of nodulation, and an increase in yield, and more preferably an increase in yield.
- the leguminous plant growth promoter according to any one of [1] to [8].
- the leguminous plant growth promoter is Preferably, plant essential nutrients, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleobases, sugars, monohydric alcohols, nonionic surfactants, food additives, microbial extracts, plant hormones, nod factors That is, it consists of lipo-chitooligosaccharide, synthetic lipo-chitooligosaccharide, chitooligosaccharide, chitinous compound, linoleic acid or its derivatives, linolenic acid or its derivatives, calicin, acyl-homoserine lactone derivative, betaine compound, and phenolic compound One or more selected from the group, More preferably, catechins or iron (III) phosphate, The use according to [10], further comprising: [12] The use according to [10] or [11], wherein the leguminous plant growth promoter is preferably an agrochemical,
- Growth promotion is preferably one or more selected from the group consisting of an increase in underground weight, an increase in the number of side buds, promotion of nodulation, and an increase in yield, and more preferably an increase in yield. Use of any one of [10] to [17].
- the glycoside is a plant essential nutrient, flavonoid, organic acid, amino acid, peptide, nucleoside, nucleotide, nucleobase, sugar, monohydric alcohol, nonionic surfactant, food additive, Microbial extract, plant hormone, nod factor ie lipo-chitooligosaccharide, synthetic lipo-chitooligosaccharide, chitooligosaccharide, chitinic compound, linoleic acid or its derivatives, linolenic acid or its derivatives, calicin, acyl-homoserine lactone derivative, One or more selected from the group consisting of betaine compounds and phenolic compounds, More preferably, catechins or iron (III) phosphate, [19] The use according to [19].
- the glycoside is contained in a pesticide, a fertilizer, a microbial material, a soil conditioner, a sowing material or a plant supplement for legumes, [19] or [20] use.
- the microbial material is preferably a rhizobial material.
- the agrochemical, fertilizer, microbial material, soil conditioner, sowing material or plant supplement contains 0.0005 to 80% by mass of the glycoside, [21] or [22] Use of.
- the cultivation base material is preferably soil, culture medium, culture medium, hydroponic solution or water.
- the cultivation base preferably contains 0.01 to 100 ppm by mass of the glycoside.
- Growth promotion is preferably any one or more selected from the group consisting of an increase in underground weight, an increase in the number of lateral buds, promotion of nodulation, and an increase in yield, and more preferably an increase in yield.
- the glycoside is Preferably, plant essential nutrients, flavonoids, organic acids, amino acids, peptides, nucleosides, nucleotides, nucleobases, sugars, monohydric alcohols, nonionic surfactants, food additives, microbial extracts, plant hormones, nod factors That is, it consists of lipo-chitooligosaccharide, synthetic lipo-chitooligosaccharide, chitooligosaccharide, chitinous compound, linoleic acid or its derivatives, linolenic acid or its derivatives, calicin, acyl-homoserine lactone derivative, betaine compound, and phenolic compound One or more selected from the group, More preferably, catechins or iron (III) phosphate, The method according to [28] or [29], which is used in combination.
- the addition of the glycoside of Soyasapogenol B to the cultivation base material includes an agrochemical, a fertilizer, a microbial material, a soil improver, and a seeding containing the glycoside as an active ingredient to the cultivation base material
- the method according to [32] comprising adding a plant material or a plant supplement.
- the application or smearing of the soyasapogenol B glycoside to the legume seeds includes an agrochemical, a fertilizer, a microbial material, a soil conditioner containing the glycoside as an active ingredient on the seeds,
- the method according to [33] comprising applying or smearing a seeding material or a plant supplement.
- the spraying, spraying or application of the glycoside of Soyasapogenol B to the legume plant body includes an agrochemical, fertilizer, or microbial material containing the glycoside as an active ingredient on the plant body
- the method according to [34] comprising spraying, spraying or applying a soil conditioner, a seeding material or a plant supplement.
- the agrochemical, fertilizer, microbial material, soil conditioner, seeding material or plant supplement contains 0.0005 to 80% by mass of the soyasapogenol B glycoside, [35] to [37 ] The method of any one of the above. [39] The method according to any one of [31] to [38], wherein, during the cultivation, the concentration of the glycoside in the cultivation substrate is 0.01 to 100 ppm by mass. . [40] The method according to any one of [31] to [39], wherein the cultivation base material for cultivation is preferably soil, culture medium, culture medium, solution for hydroponics or water.
- Growth promotion is preferably any one or more selected from the group consisting of an increase in subsurface weight, an increase in the number of side buds, promotion of nodulation, and an increase in yield, and more preferably an increase in yield.
- [42] The method according to any one of [28] to [41], wherein soil or cultivated soil that has been agglomerated with an alkali-treated lignin is preferably used as a cultivation base.
- the glycoside is preferably used in combination with a microbial cell or a microbial material.
- the glycoside of soyasapogenol B is a compound represented by the above formula (I) (in the formula (I), R is a sugar residue or a sugar chain, Preferably, it is a sugar residue selected from the group consisting of glucuronic acid, galactose, glucose, rhamnose and arabinose, or rhamnose (1 ⁇ 2) galactose (1 ⁇ 2) glucuronic acid (1 ⁇ 3), rhamnose ( 1 ⁇ 2) A sugar chain selected from the group consisting of arabinose (1 ⁇ 2) glucuronic acid (1 ⁇ 3) and glucose (1 ⁇ 2) galactose (1 ⁇ 2) glucuronic acid (1 ⁇ 3)) ; More preferably, it is at least one selected from the group consisting of soyasaponin Bb, soyasaponin Bc and soyasaponin Ba; More preferred is Soyasaponin Bb
- the legumes are: Preferably, it is at least one selected from the group consisting of soybean genus plant, kidney bean plant, chick genus plant, pea plant, lentil plant, bean plant, broad bean plant, and peanut plant, More preferably, it is at least one selected from the group consisting of soybeans, kidney beans, chickpeas, peas, lentils, pigeons, broad beans and peanuts.
- the soil is soil or culture soil that has been agglomerated using an alkali-treated lignin.
- Example 1 (Effects of soyasaponin Bb on soybean growth) After irrigating 100 mL of tap water into Leonard jar (Soil Science and Plant Nutrition, 1983, 29: 97-100) filled with about 65 g of soil (Takii hydrous cell soil TM-1, Takii Seed and Seed Co., Ltd.), soybean ( Two varieties “Midori” and Kaneko Seedling Co., Ltd.) were seeded at a depth of about 1 cm from the surface of the soil.
- soil Teakii hydrous cell soil TM-1, Takii Seed and Seed Co., Ltd.
- soybean Two varieties “Midori” and Kaneko Seedling Co., Ltd.
- soyasaponin Bb Soyasaponin I, ChromaDex, Inc.
- 50 mM TAPS buffer 50 mM TAPS buffer (pH 7.7) was dropped onto a soil covering the seeds using a micropipette.
- the soyasaponin Bb concentration in the soil was about 0.3 ppm (mass ppm, the same applies hereinafter).
- 50 mM TAPS buffer 50 mM TAPS buffer (pH 7.7) was applied in the same procedure.
- Plants were cultivated in an artificial meteorograph (LPH-350SP, Nippon Medical Instrument Co., Ltd.).
- the light conditions were light period (light intensity 130 ⁇ mol / m 2 / s) 16 hours / dark period 8 hours, temperature 26 degrees C. light period 20 degrees C., and humidity 50%.
- Water was replenished by adding an appropriate amount of tap water to the bottom of the jar as appropriate (about once every 7-10 days). 24 days after sowing, growth indices (leaf age, plant height, number of side buds, aboveground fresh weight, underground fresh weight) were measured.
- SSB represents a soyasaponin Bb application zone
- non-application zone represents a soyasaponin Bb non-application zone.
- soyasaponin Bb The leaf age and plant height were not significantly different between the soyasaponin Bb application group and the non-application group, and the values were not lowered by the addition of soyasaponin Bb (FIGS. 1 and 2).
- soyasaponin Bb since a significant increase in the underground weight (see Non-Patent Document 1), which is a growth index related to the increase in soybean seed yield, was observed, soyasaponin Bb was applied to soybean, It was expected that soybean seed yield could be increased.
- Example 2 (Effect of soyasaponin Bb on soybean growth: combined use with rhizobial materials) After irrigating 100 ml of tap water to Leonard jar filled with culture soil (Takii hydrous cell culture soil TM-1, Takii Seed and Seed Co., Ltd.), using spatula, rhizobial material ("Dr Mametaro (registered trademark)" (Idemitsu) Kosan Co., Ltd.) was sprayed on the cultivated soil in the jar so that the thickness was about 5 mm, and here two soybeans (variety “Midori”, Kaneko Seed Co., Ltd.), about 1 cm from the surface 1 mL of a 20 ⁇ M soyasaponin Bb (Soyasaponin I, ChromaDex, Inc.) solution dissolved in 50 mM TAPS buffer (pH 7.7) was dropped on a soil covering the seeds using a micropipetter.
- soyasaponin Bb Soya
- soyasaponin Bb concentration in the soil was about 0.3 ppm, only 1 mL of 50 mM TAPS buffer as a negative control, As a positive control for the promotion of formation, 1 mL of 20 ⁇ M genistein (Gen) solution was applied in the same manner (see the following for the nodulation promotion effect of genistein: Plant and Soil, 1997, 192: 141-151; Secretions and Updates in Biological Systems 27-48, Signaling and Communication in Plants 12, Springer-Verlag Berlin Heidelberg, 2012).
- + Gen represents the genistein application area
- + SSB represents the soyasaponin Bb application area.
- FIGS. 6 to 12 when a significant difference between groups is found with a significance level of less than 5% by a significant difference test between groups, different alphabets (a, b, c) are attached. It was shown that there was a significant difference between the groups. Only in the soyasaponin Bb application group, the fresh weight of the underground portion was statistically significantly increased (about 1.5 times) as compared to the non-application group (only the rhizobial material) (FIG. 10).
- Non-Patent Document 1 which is a growth index related to soybean seed yield increase, and the number of nodules also related to seed yield increase Since an increasing tendency of nodule weight (see Non-Patent Document 2) was recognized, it was expected that soybean seed yield could be increased by applying soyasaponin Bb and rhizobial material to soybean.
- Example 3 (Effect of soyasaponin Bb-containing soybean saponin preparation on soybean growth: combined use with rhizobial materials) Soybean saponin preparation containing about 10% (w / w) of soyasaponin Bb (“saponin, derived from soybean”; Wako Pure Chemical Industries, Ltd.) and rhizobial material (“Dr Mametaro (registered trademark)” (Idemitsu Kosan) In order to obtain a constant concentration (0 ppm, 50 ppm, 100 ppm or 500 ppm), the Leonard jar filled with the soil (Takiicell Culture TM-1, Takii Seed Co., Ltd.) was fed overnight from the bottom.
- + S50, + S100, and + S500 represent the soybean saponin preparations 50 ppm, 100 ppm, and 500 ppm, respectively.
- FIGS. 13 to 17 when a significant difference between groups is found with a significance level of less than 5% by a significant difference test between groups, different alphabets (a, b, c) are attached to the groups. It was shown that there was a significant difference between them. Nodule formation was not observed in soybean in the non-application area grown only with cultivation soil. When only the rhizobial material was applied, the formation of about 2 nodules per plant individual was observed.
- the number of nodules increased statistically significantly, and only the rhizobial material increased more than three times the application area (FIG. 13).
- the root nodule fresh weight per plant individual also increased statistically significantly compared to the application of only the rhizobial material (FIG. 14). The largest increase in both the number of nodules and the fresh weight of nodules was observed in the 50 ppm soybean saponin preparation.
- Non-Patent Document 2 which are growth indicators related to the increase in soybean seed yield. It was expected that the seed yield of soybean could be increased by applying the rhizobial material added with the saponin preparation to soybean.
- YM (Yeast Extract Mannitol) medium K 2 HPO 4 0.5 g, MgSO 4 .7H 2 O 0.2 g, NaCl 0.1 g, Yeast Extract 0.4 g, Mannitol 10 g, distilled water 1 L (pH 6.8) ) was added with 1.5% agar (Wako Pure Chemical Industries, Ltd.) to prepare an agar medium, and the nodule fungus Bradyrhizobium japonicum NBRC14783T was grown. The grown rhizobia was inoculated with one platinum ear in 5 mL of YM medium prepared in a 50 mL test tube, and cultured with shaking at 30 ° C.
- rhizobial culture solution was inoculated into 100 mL of YM medium having the same composition prepared in a 500 mL Sakaguchi flask, and cultured with shaking for about 72 hours.
- 1 mL of a rhizobial culture solution in which the value of the turbidity OD600 of the cells was grown to about 0.3 was inoculated dropwise from above the seeds sowed using a micropipette.
- soybean saponin 80 (soybean saponin 80”; Access One Co., Ltd.) solution dissolved in Milli-Q water was prepared, and 0.2 mL of the solution was dropped from above the seeds using a micropipette.
- soy saponin preparation corresponds to about 0.067 ppm as the concentration in the soil.
- soyasaponin concentration of group B group in the soil was determined in the soybean saponin quantified in Production Example 1 described later. Equivalent to about 0.015 ppm in terms of content in the formulation). Only the bacterial solution was dropped into the negative control (“non-application area”).
- soybean seedlings were thinned out to 1 strain per pot.
- the light conditions were set to 16 hours / dark period 8 hours.
- the temperature was 25 ° C.
- Example 5 (Effect of soyasaponin Bb-containing soybean saponin preparation on soybean yield: application by soil irrigation)
- “Fukuyutaka” purchased from Nikko Seed Seed Co., Ltd.
- saponin preparation (“soybean saponin 80”; Access One Co., Ltd.) or 5 mg of purified saponin preparation prepared in Production Example 1 described below was suspended in 100 mL of tap water and irrigated on the surface of the soil (in the medium)
- concentrations of the saponin preparation and the purified saponin preparation were about 12.5 ppm and about 1.25 ppm, respectively, and the concentration of Group B soyasaponin converted from the quantitative results in Production Example 1 (Table 7) was about 6.1 ppm and about 0. Equivalent to 9 ppm).
- non-application area As a negative control, only 100 mL of tap water was irrigated in the same manner (“non-application area”).
- rhizobial fungus rhizobium Bradyrhizobium japonicum NBRC14783T strain
- soybean seedlings were thinned out to 2 strains per pot.
- the light conditions were set to 16 hours light period / 8 hours dark period from the sowing to the 47th day and 12 hours light period / 12 hours dark period after the 48th day.
- Light conditions were set to 16 hours of light period / 8 hours of dark period from the sowing to 13th day, and 12 hours of light period / 12 hours of dark period after the 14th day.
- the temperature was 26 ° C. in the light period / 20 ° C. in the dark period, and the humidity was 50%.
- Example 7 (Effects of soyasaponin Bb-containing soybean saponin preparations on soybean yield: application by mixing soil with soil) As the soybean seeds, “Fukuyutaka” (purchased from Nikko Seed Seed Co., Ltd.) was used. Add “Mizuho Kasei Fertilizer No.
- the surface layer of several centimeters was mixed with a spatula (the concentration of the saponin preparation in the medium was about 12.5 ppm, and the concentration of group B group soyasaponin converted from the quantitative results in Production Example 1 described later (Table 7)) Equivalent to about 6.1 ppm).
- the saponin formulation was not added to the negative control ("Non-application area").
- Three seeds were sown at a depth of about 1 cm from the surface of the soil. From the seed sowed, 1 mL of a rhizobial fungus (rhizobium Bradyrhizobium japonicum NBRC14783T strain) prepared in the same manner as in Example 4 was dropped using a micropipetter.
- soybean seedlings were thinned out to 1 strain per pot.
- Light conditions were set to 16 hours of light period / 8 hours of dark period from the sowing to 13th day, and 12 hours of light period / 12 hours of dark period after the 14th day.
- the temperature was 26 ° C. in the light period / 20 ° C. in the dark period, and the humidity was 50%.
- the fresh weight of the saponin preparation 50 mg application group showed an increase of about 20%, although there was no significant difference (Tukey-Kramer method) compared to the non-application group (FIG. 23).
- Table 5 shows the yield per 10 ares calculated from the dry weight of the seeds.
- the yield of the saponin preparation 50 mg application group increased by 17% on average compared to the non-application group.
- Example 8 (Effect of combined use of soyasaponin Bb-containing soybean saponin preparation and soil improver alkali-treated lignin on soybean yield) Using the soil that had been subjected to agglomeration treatment, the yield increase of soybean was examined when a saponin preparation was applied.
- As the soybean seed “Yuagagarusume” (Kaneko Seedling Co., Ltd.) was used.
- Example 9 (Effect of combined use of soyasaponin Bb-containing soybean saponin preparation and catechin on soybean growth) After supplying water from the bottom to Leonard jar (Soil Science and Plant Nutrition, 1983, 29: 97-100) filled with about 65 g of soil (Takii hydrous cell culture soil TM-1, Takii Seed and Seed Co., Ltd.), soybean (Cultivar “Fukuyutaka”, Nikko Seed Seed Co., Ltd.) was seeded one by one at a depth of about 1 cm from the surface.
- YM (Yeast Extract Mannitol) medium K 2 HPO 4 0.5 g, MgSO 4 .7H 2 O 0.2 g, NaCl 0.1 g, Yeast Extract 0.4 g, Mannitol 10 g, distilled water 1 L (pH 6.8)
- Nodule Mycobacterium Bradyrhizobium japonicum NBRC14783T is grown on an agar medium prepared by adding 1.5% agar (Wako Pure Chemical Industries, Ltd.) to a 5 mL YM medium prepared in a 50 mL test tube. Platinum ears were inoculated and cultured with shaking at 30 ° C. and 250 rpm for 24 hours.
- Rhizobium culture solution was inoculated into 100 mL of YM medium prepared in a 500 mL Sakaguchi flask, and cultured with shaking at 30 ° C. and 120 rpm, and the bacteria were grown to an OD600 of about 0.3. 1 mL of the obtained rhizobial culture solution was inoculated dropwise from above the seeds sowed.
- Plants were cultivated using an artificial meteorograph (LPH-411SP, Nippon Medical Instrument Co., Ltd.).
- the light conditions were light period (light intensity 130 ⁇ mol / m 2 / s) 12 hours / dark period 12 hours, temperature 25 ° C./dark period 20 ° C., and humidity 50%.
- the measurement result of the underground dry weight is shown in FIG.
- FIG. 24 when a significant difference is recognized between the mean values with a significance level of less than 5% by the significance test (Dunnett method) of the section, different alphabets (a, b) are attached to the sections. It showed that there was a significant difference. Compared to the non-application area, the saponin area and the catechin area increased the dry weight of the underground (about 22% and about 45% increase, respectively). Furthermore, the underground dry weight in the combination group showed a significant increase (an increase of about 64%, a significance level of 5% or less) compared to the non-application group, and increased more than the saponin group and the catechin group.
- saponin preparation (“soybean saponin 80”; Access One Co., Ltd.), 0.75 mg of iron (III) phosphate tetrahydrate (Junsei Co., Ltd.), and a mixture thereof were each added to 100 mL of tap water.
- saponin group “soybean saponin 80”; Access One Co., Ltd.)
- iron (III) phosphate tetrahydrate (Junsei Co., Ltd.)
- concentration of the saponin preparation in the combination group and saponin group was about 12.5 ppm
- concentration of group B group soyasaponin converted from the quantitative results in Example 1 corresponds to about 6.1 ppm).
- the persimmon fresh weight increased by about 10% in the saponin group and iron phosphate group compared to the non-application group, and in the combination group significantly increased compared to the non-application group (about 12% increase, significance level of 5% or less) (FIG. 25).
- persimmon fresh weight significantly increased (about 10% increase, significant level of 5% or less) in the iron phosphate group and the combination group compared to the non-application group (FIG. 26).
- Example 11 (Effect of combined use of soyasaponin Bb-containing soybean saponin preparation and iron phosphate on the yield of soybeans grown in non-cultivated soil) Soybean seeds were “green cocoon” (Kaneko Seedling Co., Ltd.). Fill the Wagner pot of 1/5000 are filled with about 4L of soil (soil from non-cultivated land that has not been fertilized, etc., collected from the Tochigi Plant of Kao Corporation) and about 1cm deep from the surface of the soil Three seeds were sown.
- Combination zone the concentration of the saponin preparation in the soil is about 12.5 ppm and corresponds to about 6.1 ppm as the concentration of group B group soyasaponin converted from the quantitative results in Production Example 1 (Table 7).
- As a control only 100 mL of tap water was irrigated in the same manner (“non-application area”).
- the light conditions were set to 16 hours light period / 8 hours dark period. The temperature was 26 ° C.
- the dry weight per pot in the combination group increased by about 30% although there was no significant difference from the non-application group (FIG. 27).
- the combination group significantly increased compared to the non-application group (about 240% increase, significance level of 5% or less) (FIG. 28).
- the seedlings were removed from the soil, and the roots were washed with running water and filled with 50 mL of Hoagland nutrient solution (prepared using Hoagland Modified Basal Salt Mixture (PhytoTechnology Laboratories)) diluted to a predetermined 5-fold concentration.
- Hoagland nutrient solution prepared using Hoagland Modified Basal Salt Mixture (PhytoTechnology Laboratories)
- One individual was transferred to the screw tube, Hoaglan in the screw tube.
- saponin preparation (“soybean saponin 80”; Access One Co., Ltd.) at a final concentration of 0 ppm (“non-application area”), 10 ppm (“10 ppm application area”) or 100 ppm (“100 ppm application area”)
- concentration of group B group soyasaponin in the nutrient solution corresponds to 0 ppm, about 4.9 ppm, and about 49.1 ppm as converted concentrations from the quantitative results in Production Example 1 (Table 7)).
- kidney beans and peas were prepared in 5 samples, and the plants were cultivated in an artificial meteorometer, the light conditions were 16 hours light period / 8 hours dark period, the temperature was 26 ° C light period / 20 ° C dark period, and the humidity was The nutrient solution was replaced with a new one containing no saponin preparation every 7 days, 14 days after the transition to the nutrient solution, the fresh weight of the above ground part and the fresh part of the underground part were measured. Using the Dunnett's test to multiple testing of the physical section.
- both the 10 ppm application area of the saponin preparation and the fresh underground weight of the 100 ppm application area increased about 1.3 times as compared to the non-application area (FIG. 29).
- the fresh underground weight increased significantly (about 20%, significant level 5% or less) in the saponin preparation 10 ppm application area compared to the non-application area, and an increase of about 15% was observed even in the 100 ppm application area.
- FIG. 30 Regarding the fresh weight of the aerial part, an increase of 18% or more was observed in the pea saponin application area compared to the non-application area (FIG. 30).
- Example 13 (Growth promotion effect of soybean and chickpea by foliar application of saponin preparation)
- a seedling box vertical 270 mm ⁇ width 345 mm ⁇ height 75 mm
- soybean variety “Fukuyutaka”; Nikko seedling
- seeds of chickpea variety unknown (fog type); purchased from Nikko Seed Seed Co., Ltd.
- the soybean seeds are arranged at an equal interval of about 1 cm and 5 vertical x 9 horizontal, and the chickpea seed is arranged at an equal interval of about 1 cm and 4 vertical x 9 horizontal. It was made to become.
- the seedling box was installed on the bat, and water was supplied by appropriately supplying tap water to the bat. Cultivation was carried out in an artificial meteorograph, light conditions were set to 16 hours light period / 8 hours dark period, temperature 26 ° C./dark period 20 ° C., and humidity 50%.
- For soybean a group of 5 vertical x 3 horizontal plants was taken as one unit.
- chickpeas a group of 4 vertical and 3 horizontal plants was taken as one unit.
- saponin preparation ("soybean saponin 80"; Access One Co., Ltd.) in an aqueous solution containing 0.05% by mass of approach BI (Kao Co., Ltd.) as a treatment liquid as a treatment liquid.
- BI Kao Co., Ltd.
- each of these treatment solutions was sprayed (about 5 mL / unit) on the above-ground part of each unit using a spray bottle 13 days after sowing of each plant species (the concentration of the applied saponin preparation is included in the soil of each unit) If so, the concentration of group B group soyasaponin converted from the quantitative results in Production Example 1 (Table 7) is equivalent to about 0.018 ppm and 0.18 ppm).
- a paper partition was set up between the units to prevent the treatment liquid from scattering into different units.
- the appearance of the roots extending from the bottom of the seedling box of each plant species was photographed. From the photographed image, the area (rooting) of the root extension region in each application section was analyzed and digitized as follows using the open source image analysis software ImageJ.
- the captured image was RGB-divided by the “Split Channels” function of ImageJ.
- the analysis area (vertical seedling box vertical (270 mm) and horizontal (345 mm)) in the image is divided into equal intervals by lines with a width of 3 pixels, and formed at the intersection of each line. A total of 1536 sections were divided. The length of 1 pixel in the image corresponds to 0.013 cm for the soybean image and 0.012 cm for the chickpea image.
- the brightness threshold (“Threshold” item) on the software for identifying root parts was set to 106-255 for soybean and 116-255 for chickpeas.
- the threshold value was set in accordance with a conventional method while visually confirming that an appropriate range was selected on the image (reference document: Tajima, Nemoto no 23 (3): 75-81 (2014)).
- the area of the root extension region is measured by executing “Analyze Particles”. In this case, the “Size” item in the software is 0 to 100000 (cm 2 ), and the “Circularity” item is 0.01. -1.00. From the measurement result, the relative value when the rooting in the non-application area was set to 1 was obtained.
- the fresh weight of the above-ground part significantly increased (about 7% increase, significant level of 5% or less) in the 10 ppm application group in comparison with the non-application group (FIG. 31).
- the above-ground fresh weight significantly increased in the 10 ppm application zone and the 100 ppm application zone as compared to the non-application zone (an increase of about 30%, a significance level of 5% or less and 1% or less, respectively) (FIG. 32).
- rooting an increase in rooting was observed in the saponin application group compared to the non-application group in both soybean and chickpea (FIGS. 33A and B).
- the rooting increase is about 2.3 times in the 10 ppm application zone and about 3.1 times in the 100 ppm application zone compared to the non-application zone. (Fig. 34), and also in chickpeas, an increase in rooting of about 2.1 times in the 10 ppm application zone and about 2.9 times in the 100 ppm application zone was observed (Fig. 34). 35).
- Production Example 1 1) Purification of saponin preparation 4.9 g of a soybean saponin preparation ("soybean saponin 80"; Access One Co., Ltd.) was dissolved in 300 mL of 40 v / v% ethanol, and centrifuged to collect the supernatant.
- group B group soyasaponin was then purified from the 60 v / v% ethanol fraction. 0.87 g of the freeze-dried 60 v / v% ethanol fraction was redissolved in 200 mL of 60 v / v% ethanol, and 1.0 g of activated carbon (Shirakaba P, Osaka Gas Chemical) was added thereto, followed by stirring with a stirrer for 1 hour. After filtration through a PTFE filter, the filtrate was concentrated under reduced pressure, and then freeze-dried to obtain a powdered purified saponin preparation. The soyasaponin content of Group B in the soybean saponin preparation and the obtained purified saponin preparation was quantified by the following procedure.
- the eluent used was A: 0.1 v / v% formic acid water, B: acetonitrile, and the gradient conditions were 0 min to 1 min (10 v / v% B) ⁇ 1 min to 7 min (1 v / v% B to 47 0.5 v / v% B) ⁇ 7 to 9 minutes (47.5 v / v% B to 85 v / v% B) ⁇ 9 to 9.01 minutes (85 v / v% B to 100 v / v% B) ⁇ 9.01 to 10 minutes (100 v / v% B) ⁇ 10 minutes to 10.01 minutes (100 v / v% B to 10 v / v% B) ⁇ 10.01 minutes to 11 minutes (10 v / v% B) It was.
- the flow rate was 0.5 mL / min.
- the MRM method multiple reaction monitoring
- Soyasaponins I, II and V contained in each sample were quantified from a calibration curve.
- Table 7 shows the total amount (mass%) of soyasaponin of group B in each preparation calculated from the quantitative values.
- an alkali-treated lignin (lignin degradation product) serving as a soil aggregating agent was produced by the following steps 1 and 2.
- lignin degradation product lignin degradation product
- steps 1 and 2 As herbaceous biomass, sugarcane bagasse was put in a 30 g glass bottle as a dry mass, and a 1.6 mass% sodium hydroxide aqueous solution was added so that the solid content was 10 mass%. The glass bottle was heated in an autoclave at 95 ° C. for 6 hours to obtain a reaction product.
- Step 2 The reaction product obtained in Step 1 was filtered under reduced pressure using a 400 mesh SUS mesh and Nutsche. The residue was washed with 300 mL of ion exchange water at 90 ° C. The filtrate and the washing solution were collected, and the pH was adjusted to 4 with 1.0 M hydrochloric acid to obtain a suspension containing a lignin decomposition product.
- step 2 The suspension obtained in step 2 was centrifuged. Centrifugation was performed under the conditions of 10000 rpm for 20 minutes using “himac CR 20G III” manufactured by Hitachi Koki Co., Ltd. After centrifugation, the supernatant was removed, and 300 mL of ion exchange water was added and stirred. Then, it centrifuged again under the same conditions as the above, and washed with water. Washing with water was performed twice, and the resulting precipitate was freeze-dried to obtain a powdery alkali-treated lignin (lignin decomposition product).
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Abstract
Description
(特許文献2)特開昭61-15806号公報
(特許文献3)特開平8-23963号公報
(非特許文献1)日本育種学会・日本作物学会北海道談話会会報,1992, (31):64
(非特許文献2)作物研究所研究報告, 2007, (8):49-108
(非特許文献3)種子の科学とバイオテクノロジー, 種子生理生化学研究会(編集), 学会出版センター, 2009, pp.106-112
(非特許文献4)Botanical Bulletin of Academia Sinica, 1995, 36(1):9-18
(非特許文献5)Advances in Plant Glycosides, Chemistry and Biology, Volume 6, Elsevier Science, 1999, pp.105-130
(非特許文献6)Plant and Soil, 1987, 98(1):67-80
(非特許文献7)Physiol Plantarum, 1995, 93(4):785-789
(非特許文献8)Biologically Active Natural Products: Agrochemicals, CRC Press, 1999, pp. 248-272
(非特許文献9)Isoprenoid Synthesis in Plants and Microorganisms: New Concepts and Experimental Approaches, Springer, 2013, pp.405-424
(非特許文献10)Phytochem Rev, 2013, (12):877-893
(非特許文献11)大豆たん白質研究会会誌, 1994, (15):36-40
さらなる一実施形態において、本発明は、ソヤサポゲノールBの配糖体であって、該ソヤサポゲノールBのC-22位がヒドロキシ基であり、かつ該ソヤサポゲノールBのC-3位ヒドロキシ基に糖が結合している配糖体を有効成分として用いるマメ科植物の生育促進方法を提供する。
別の一実施形態において、本発明は、マメ科植物生育促進剤の製造のためのグループB群のソヤサポニンの使用を提供する。
別の一実施形態において、本発明は、マメ科植物生育促進のためのグループB群のソヤサポニンの使用を提供する。
別の一実施形態において、本発明は、マメ科植物生育促進に使用するためのグループB群のソヤサポニンを提供する。
別の一実施形態において、本発明は、グループB群のソヤサポニンを用いたマメ科植物の生育促進方法を提供する。
本発明において、該生育促進とは、好ましくは地下部の重量の増加、側芽数の増加、収量の増加、および根粒形成の促進からなる群より選択されるいずれか1以上をいい、より好ましくは収量の増加、特に一株または単位面積当たり種子の総重量を増加させることをいう。
・ダイズ属植物用としては、Bradyrhizobium属菌およびEnsifer (Sinorhizobium)属菌からなる群より選択される1種以上が好ましく、Bradyrhizobium属菌がより好ましい。Bradyrhizobium属菌としては、Bradyrhizobium japonicum、Bradyrhizobium diazoefficiensおよびBradyrhizobium elkaniiからなる群より選択される1種以上が好ましく、Bradyrhizobium japonicumおよびBradyrhizobium diazoefficiensからなる群より選択される1種以上がより好ましい。Ensifer (Sinorhizobium)属菌としてはEnsifer (Sinorhizobium) frediiが好ましい;
・インゲンマメ属植物、エンドウ属植物、ソラマメ属植物、ヒラマメ属植物、およびキマメ属植物用としては、Rhizobium属菌が好ましい。Rhizobium属菌としては、Rhizobium leguminosarum、Rhizobium gallicumおよびRhizobium giardiniiからなる群より選択される1種以上が好ましく、Rhizobium leguminosarumがより好ましい;
・ヒヨコマメ属植物用としては、Mesorhizobium属が好ましく、Mesorhizobium属としては、Mesorhizobium ciceriおよびMesorhizobium mediterraneumからなる群より選択される1種以上が好ましく、Mesorhizobium ciceriがより好ましい;
・ラッカセイ属植物用としては、Bradyrhizobium属菌が好ましい。好ましいBradyrhizobium属菌の種はダイズ属植物の場合と同様である。
〔2〕好ましくは植物の必須栄養素、フラボノイド、有機酸、アミノ酸、ペプチド、ヌクレオシド、ヌクレオチド、核酸塩基、糖、1価アルコール、非イオン性界面活性剤、食品添加物、微生物抽出物、植物ホルモン、nod因子すなわちリポ-キトオリゴ糖、合成リポ-キトオリゴ糖、キトオリゴ糖、キチン性化合物、リノール酸またはその誘導体類、リノレン酸またはその誘導体類、カリキン、アシル-ホモセリンラクトン誘導体、ベタイン化合物、およびフェノール類化合物からなる群より選択される1つ以上、
より好ましくは、カテキン類またはリン酸鉄(III)、
をさらに含有する、〔1〕記載のマメ科植物生育促進剤。
〔3〕好ましくは、マメ科植物用の農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントの形態である、〔1〕または〔2〕記載のマメ科植物生育促進剤。
〔4〕好ましくは、前記微生物資材が根粒菌資材である、〔3〕記載のマメ科植物生育促進剤。
〔5〕好ましくは前記配糖体を0.0005~80質量%含有する、〔3〕または〔4〕記載のマメ科植物生育促進剤。
〔6〕好ましくは、マメ科植物用の栽培基材である、〔1〕または〔2〕記載のマメ科植物生育促進剤。
〔7〕好ましくは、前記栽培基材が土壌、培土、培地、養液栽培用溶液または水である、〔6〕記載のマメ科植物生育促進剤。
〔8〕好ましくは前記配糖体を0.01~100質量ppm含有する、〔6〕または〔7〕記載のマメ科植物生育促進剤。
〔9〕生育促進が、好ましくは地下部重の増加、側芽数の増加、根粒形成の促進および収量増加からなる群より選択されるいずれか1以上であり、より好ましくは収量増加である、〔1〕~〔8〕のいずれか1項記載のマメ科植物生育促進剤。
〔11〕前記マメ科植物生育促進剤が、
好ましくは、植物の必須栄養素、フラボノイド、有機酸、アミノ酸、ペプチド、ヌクレオシド、ヌクレオチド、核酸塩基、糖、1価アルコール、非イオン性界面活性剤、食品添加物、微生物抽出物、植物ホルモン、nod因子すなわちリポ-キトオリゴ糖、合成リポ-キトオリゴ糖、キトオリゴ糖、キチン性化合物、リノール酸またはその誘導体類、リノレン酸またはその誘導体類、カリキン、アシル-ホモセリンラクトン誘導体、ベタイン化合物、およびフェノール類化合物からなる群より選択される1つ以上、
より好ましくは、カテキン類またはリン酸鉄(III)、
をさらに含有する、〔10〕記載の使用。
〔12〕好ましくは、前記マメ科植物生育促進剤がマメ科植物用の農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントである、〔10〕または〔11〕記載の使用。
〔13〕好ましくは、前記微生物資材が根粒菌資材である、〔12〕記載の使用。
〔14〕好ましくは、前記マメ科植物生育促進剤が前記配糖体を0.0005~80質量%含有する、〔12〕または〔13〕記載の使用。
〔15〕好ましくは、前記マメ科植物生育促進剤がマメ科植物用の栽培基材である、〔10〕または〔11〕記載の使用。
〔16〕好ましくは、前記栽培基材が土壌、培土、培地、養液栽培用溶液または水である、〔15〕記載の使用。
〔17〕好ましくは、前記マメ科植物生育促進剤が前記配糖体を0.01~100質量ppm含有する、〔15〕または〔16〕記載の使用。
〔18〕生育促進が、好ましくは地下部重の増加、側芽数の増加、根粒形成の促進および収量増加からなる群より選択されるいずれか1以上であり、より好ましくは収量増加である、〔10〕~〔17〕のいずれか1項記載の使用。
〔20〕好ましくは、前記配糖体が、植物の必須栄養素、フラボノイド、有機酸、アミノ酸、ペプチド、ヌクレオシド、ヌクレオチド、核酸塩基、糖、1価アルコール、非イオン性界面活性剤、食品添加物、微生物抽出物、植物ホルモン、nod因子すなわちリポ-キトオリゴ糖、合成リポ-キトオリゴ糖、キトオリゴ糖、キチン性化合物、リノール酸またはその誘導体類、リノレン酸またはその誘導体類、カリキン、アシル-ホモセリンラクトン誘導体、ベタイン化合物、およびフェノール類化合物からなる群より選択される1つ以上、
より好ましくは、カテキン類またはリン酸鉄(III)、
と併用される、〔19〕記載の使用。
〔21〕好ましくは、前記配糖体が、マメ科植物用の農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントに含有されている、〔19〕または〔20〕記載の使用。
〔22〕好ましくは、前記微生物資材が根粒菌資材である、〔21〕記載の使用。
〔23〕好ましくは、前記農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントが、前記配糖体を0.0005~80質量%含有する、〔21〕または〔22〕記載の使用。
〔24〕好ましくは、前記配糖体がマメ科植物用の栽培基材に含有されている、〔19〕または〔20〕記載の使用。
〔25〕好ましくは、前記栽培基材が土壌、培土、培地、養液栽培用溶液または水である、〔24〕記載の使用。
〔26〕好ましくは、前記栽培基材が前記配糖体を0.01~100質量ppm含有する、〔24〕または〔25〕記載の使用。
〔27〕生育促進が、好ましくは地下部重の増加、側芽数の増加、根粒形成の促進および収量増加からなる群より選択されるいずれか1以上であり、より好ましくは収量増加である、〔19〕~〔26〕のいずれか1項記載の使用。
〔29〕ソヤサポゲノールBの配糖体であって、該ソヤサポゲノールBのC-22位がヒドロキシ基であり、かつ該ソヤサポゲノールBのC-3位ヒドロキシ基に糖が結合している配糖体を有効成分として用いる、マメ科植物の生育促進方法。
〔30〕前記配糖体が、
好ましくは、植物の必須栄養素、フラボノイド、有機酸、アミノ酸、ペプチド、ヌクレオシド、ヌクレオチド、核酸塩基、糖、1価アルコール、非イオン性界面活性剤、食品添加物、微生物抽出物、植物ホルモン、nod因子すなわちリポ-キトオリゴ糖、合成リポ-キトオリゴ糖、キトオリゴ糖、キチン性化合物、リノール酸またはその誘導体類、リノレン酸またはその誘導体類、カリキン、アシル-ホモセリンラクトン誘導体、ベタイン化合物、およびフェノール類化合物からなる群より選択される1つ以上、
より好ましくは、カテキン類またはリン酸鉄(III)、
と併用される、〔28〕または〔29〕記載の方法。
〔31〕好ましくは、前記マメ科植物を前記ソヤサポゲノールBの配糖体とともに栽培することを含む、〔28〕~〔30〕のいずれか1項記載の方法。
〔32〕好ましくは、
前記ソヤサポゲノールBの配糖体を栽培基材に有効成分として添加すること、および、
得られた該配糖体を添加した栽培基材で前記マメ科植物を栽培すること、
を含む、〔31〕記載の方法。
〔33〕好ましくは、
前記ソヤサポゲノールBの配糖体を播種前のマメ科植物種子に有効成分として塗布または塗抹すること、および、
該種子を栽培基材で栽培すること、
を含む、〔31〕記載の方法。
〔34〕好ましくは、
前記ソヤサポゲノールBの配糖体をマメ科植物の植物体に有効成分として散布、噴霧または塗布すること、および、
該植物体を栽培基材で栽培すること、
を含む、〔31〕記載の方法。
〔35〕好ましくは、前記栽培基材への前記ソヤサポゲノールBの配糖体の添加が、該栽培基材へ該配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントを添加することを含む、〔32〕記載の方法。
〔36〕好ましくは、前記マメ科植物種子への前記ソヤサポゲノールBの配糖体の塗布または塗抹が、該種子へ該配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントを塗布または塗抹することを含む、〔33〕記載の方法。
〔37〕好ましくは、前記マメ科植物の植物体への前記ソヤサポゲノールBの配糖体の散布、噴霧または塗布が、該植物体へ該配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントを散布、噴霧または塗布することを含む、〔34〕記載の方法。
〔38〕好ましくは、前記農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントが前記ソヤサポゲノールBの配糖体を0.0005~80質量%含有する、〔35〕~〔37〕のいずれか1項記載の方法。
〔39〕好ましくは、前記栽培の際に、前記栽培基材中における前記配糖体の濃度が0.01~100質量ppmである、〔31〕~〔38〕のいずれか1項記載の方法。
〔40〕好ましくは、前記栽培の栽培基材が土壌、培土、培地、養液栽培用溶液または水である、〔31〕~〔39〕のいずれか1項記載の方法。
〔41〕生育促進が、好ましくは地下部重の増加、側芽数の増加、根粒形成の促進および収量増加からなる群より選択されるいずれか1以上であり、より好ましくは収量増加である、〔29〕~〔40〕のいずれか1項記載の方法。
〔42〕好ましくは、栽培基材として、アルカリ処理リグニンを用いて団粒化処理した土壌もしくは培土を用いる、〔28〕~〔41〕のいずれか1項記載の方法。
〔43〕好ましくは、前記配糖体が微生物菌体もしくは微生物資材と併用される、〔28〕~〔42〕のいずれか1項記載の方法。
前記ソヤサポゲノールBの配糖体は、
好ましくは、上記式(I)で示される化合物であり
(該式(I)中、Rは
糖残基もしくは糖鎖であり、
好ましくは、グルクロン酸、ガラクトース、グルコース、ラムノースおよびアラビノースからなる群より選択される糖残基であるか、またはラムノース(1→2)ガラクトース(1→2)グルクロン酸(1→3)、ラムノース(1→2)アラビノース(1→2)グルクロン酸(1→3)、およびグルコース(1→2)ガラクトース(1→2)グルクロン酸(1→3)からなる群より選択される糖鎖である);
より好ましくは、ソヤサポニンBb、ソヤサポニンBcおよびソヤサポニンBaからなる群より選択される少なくとも1種であり;
さらに好ましくはソヤサポニンBbである。
〔45〕前記〔1〕~〔44〕のいずれか1項において、
前記マメ科植物は、
好ましくは、ダイズ属植物、インゲンマメ属植物、ヒヨコマメ属植物、エンドウ属植物、ヒラマメ属植物、キマメ属植物、ソラマメ属植物、およびラッカセイ属植物からなる群から選択される少なくとも1種であり、
より好ましくは、ダイズ、インゲンマメ、ヒヨコマメ、エンドウ、レンズマメ、キマメ、ソラマメおよびラッカセイからなる群から選択される少なくとも1種である。
〔46〕前記〔7〕、〔16〕、〔25〕および〔40〕のいずれか1項において、好ましくは、前記土壌はアルカリ処理リグニンを用いて団粒化処理した土壌もしくは培土である。
(ソヤサポニンBbがダイズ生育に与える効果)
約65gの培土(タキイ含水セル培土TM-1、タキイ種苗(株))を充填したLeonard jar(Soil Science and Plant Nutrition,1983,29:97-100)に水道水を100mL灌注したのち、ダイズ(品種「緑碧」、カネコ種苗(株))を2粒ずつ、培土表面から約1cmの深さに埋め込んで播種した。50mM TAPSバッファー(pH7.7)に溶解した20μMのソヤサポニンBb(ソヤサポニンI、ChromaDex,Inc.)溶液1mLを、種子を覆う培土の上にマイクロピペッターを用いて滴下した。該培土中のソヤサポニンBb濃度は約0.3ppm(質量ppm、以下同じ)であった。対照(ソヤサポニンBb非施用区)として、50mM TAPSバッファー(pH7.7)を同様の手順で施用した。
(ソヤサポニンBbがダイズ生育に与える効果:根粒菌資材併用)
培土(タキイ含水セル培土TM-1、タキイ種苗(株))を充填したLeonard jarに水道水を100mL灌注したのち、スパーテルを用いて、根粒菌資材(「Dr豆太郎(登録商標)」(出光興産(株))をjar中の培土上に厚さが5mm程度となるよう散布した。ここにダイズ(品種「緑碧」、カネコ種苗(株))を2粒ずつ、培土表面から約1cmの深さに埋め込んで播種した。50mM TAPSバッファー(pH7.7)に溶解した20μMのソヤサポニンBb(ソヤサポニンI、ChromaDex,Inc.)溶液1mLを、種子を覆う培土の上にマイクロピペッターを用いて滴下した。該培土中のソヤサポニンBb濃度は約0.3ppmであった。陰性対照として50mM TAPSバッファー 1mLのみを、また根粒形成促進の陽性対照として、20μMのゲニステイン(Gen)溶液1mLを同様に施用した(ゲニステインの根粒形成促進作用については以下を参照:Plant and Soil,1997,192:141-151;Secretions and Exudates in Biological Systems 27-48,Signaling and Communication in Plants 12,Springer-Verlag Berlin Heidelberg,2012)。
(ソヤサポニンBb含有大豆サポニン製剤がダイズ生育に与える効果:根粒菌資材併用)
ソヤサポニンBbを約10%(w/w)含有する大豆サポニン製剤(「サポニン,大豆由来」;和光純薬工業(株))を、根粒菌資材(「Dr豆太郎(登録商標)」(出光興産(株))中に一定濃度(0ppm、50ppm、100ppmまたは500ppm)となるように添加した。培土(タキイセル培土TM-1、タキイ種苗(株))を充填したLeonard jarに底面から一晩給水させたのち、大豆サポニン製剤を添加した根粒菌資材約2g(湿重量)を、jar中の培土上に均一な厚さとなるよう撒布した。該培土中のソヤサポニンBb濃度は0.15~1.5ppmであった。対照(非施用区)として培土のみのjarを準備した。ここにダイズ(品種「湯あがり娘」、カネコ種苗(株))を1粒ずつ、表面から約1cm以内の深さに埋め込んで播種した。
(ソヤサポニンBb含有サポニン製剤の生育促進効果)
ダイズ種子は「フクユタカ」(日光種苗(株)より購入)を使用した。10cm角の深型ポットに培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約1.5Lを充填し、培土表面から約1cmの深さに種子を3粒ずつ播種した。
(ソヤサポニンBb含有大豆サポニン製剤がダイズ収量に与える効果:土壌灌水による施用)
ダイズ種子は「フクユタカ」(日光種苗(株)より購入)を使用した。5000分の1アールのWagnerポットに培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約4Lを充填し、培土表面から約1cmの深さに種子を3粒ずつ播種した。サポニン製剤(「大豆サポニン80」;(株)アクセスワン)50mgまたは後述の製造例1にて作製した精製サポニン製剤5mgを、それぞれ100mLの水道水に懸濁して培土表面に灌注した(該培土中、サポニン製剤および精製サポニン製剤の濃度はそれぞれ約12.5ppmおよび約1.25ppm、かつ製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約6.1ppmおよび約0.9ppmに相当)。陰性対照として水道水100mLのみを同様に灌注した(「非施用区」)。播種した種子の上から、実施例4と同様に調製した根粒菌(根粒菌Bradyrhizobium japonicum NBRC14783T株)の菌液1mLを、マイクロピペッターを用いて滴下した。出芽後、ダイズ幼苗をポット当たり2株になるよう間引いた。光条件は播種から47日目までは明期16時間/暗期8時間、48日目以降は明期12時間/暗期12時間に設定した。温度は明期26℃/暗期20℃、湿度は50%とした。水の補給は、2~4日に1回、水道水を適量灌注することで行った。播種から126日後に、ポット当たりの子実数および重量を測定した(n=5)。
(ソヤサポニンBb含有大豆サポニン製剤がダイズ収量に与える効果:種子粉衣による施用)
ダイズ種子は「緑碧」(カネコ種苗(株))を使用した。5000分の1アールのWagner ポットに培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約4Lを充填し、培土表面から約1cmの深さに種子を3粒ずつ播種した。播種の際、サポニン製剤の粉体(「大豆サポニン80」;(株)アクセスワン)25mgまたは50mgを、直接種子の上に施用した(該培土中、各サポニン製剤の濃度はそれぞれ約6.25ppmおよび約12.5ppm、かつ後述の製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約3.1ppmおよび約6.1ppmに相当)。陰性対照にはサポニン製剤を添加しなかった(「非施用区」)。播種した種子の上から、実施例4と同様に調製した根粒菌(根粒菌Bradyrhizobium japonicum NBRC14783T株)の菌液1mLを、マイクロピペッターを用いて滴下した。出芽後、ダイズ幼苗をポット当たり1株になるよう間引いた。光条件は播種から13日目までは明期16時間/暗期8時間、14日目以降は明期12時間/暗期12時間に設定した。温度は明期26℃/暗期20℃、湿度は50%とした。水の補給は、2~4日に1回、水道水を適量灌注することで行った。播種から73日後に、ポット当たりの子実数および重量を測定した(n=10)。
(ソヤサポニンBb含有大豆サポニン製剤がダイズ収量に与える効果:田土への土壌混和による施用)
ダイズ種子は「フクユタカ」(日光種苗(株)より購入)を使用した。5000分の1アールのWagnerポットに、「みづほ化成肥料8号」(サンアグロ(株))と「グッドリンカリ肥料」((株)ことぶき門司工場)を重量比7:5で添加してN:P:K=3.5:6:6(kg/10アール換算)に調整した荒木田土約4Lを充填し、次いでサポニン製剤(「大豆サポニン80」;(株)アクセスワン)50mgを直接土壌表面に添加したのち、表層数cmをスパーテルで混和した(該培土中、サポニン製剤の濃度は約12.5ppm、かつ後述の製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約6.1ppmに相当)。陰性対照にはサポニン製剤を添加しなかった(「非施用区」)。培土表面から約1cmの深さに、種子を3粒ずつ播種した。播種した種子の上から、実施例4と同様に調製した根粒菌(根粒菌Bradyrhizobium japonicum NBRC14783T株)の菌液1mLを、マイクロピペッターを用いて滴下した。出芽後、ダイズ幼苗をポット当たり1株になるよう間引いた。光条件は播種から13日目までは明期16時間/暗期8時間、14日目以降は明期12時間/暗期12時間に設定した。温度は明期26℃/暗期20℃、湿度は50%とした。水の補給は、2~4日に1回、水道水を適量灌注することで行った。播種から90日後に、ポット当たりの子実の重量を測定した(n=6)。
(ソヤサポニンBb含有大豆サポニン製剤と土壌改良剤アルカリ処理リグニンの併用がダイズ収量に与える効果)
団粒化処理を行った土壌を使用し、加えてサポニン製剤を施用した際のダイズの収量増加性を調べた。ダイズ種子は「湯あがり娘」(カネコ種苗(株))を使用した。栽培土壌には、荒木田土とベントナイトを95:5の比率となるように混合しかつ「みづほ化成肥料8号」(サンアグロ(株))を添加してN:P:K=6:6:6(kg/10アール換算)に調整した土壌を使用した。当該土壌に対して、土壌改良作用を有するアルカリ処理リグニン(後述の製造例2(特開2017-190448号公報の明細書における製造例1の工程中の一部を省略した方法)に従って製造されたもの、以下ALと略記)を0.05質量%添加し、撹拌混合して土壌を団粒化した。当該団粒化した土壌(+AL土壌)と、ALによる団粒化処理を行わなかった土壌(-AL土壌)を、それぞれ18cm径のポリポットに約2L充填した。それぞれの土壌の表面から約1cmの深さに、ダイズ種子を2粒播種した。さらに、+AL土壌に対してのみ、サポニン製剤(「大豆サポニン80」;(株)アクセスワン)50mgを直接土壌表面に添加し、表層数cmを混和した(該土壌中、サポニン製剤の濃度は約25ppm、かつ後述の製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約12.3ppmに相当)。露地の自然光下で栽培し、給水は水道水を用いて、各ポットが均等になるように適宜行った。播種から14日後に2株出芽しているものに関しては間引き作業を行い、1ポットあたり1株になるように調整を行った。播種から111日後に、株当たりの莢の数を測定した(n=8)。
(ソヤサポニンBb含有大豆サポニン製剤およびカテキンの併用がダイズ生育に与える効果)
約65gの培土(タキイ含水セル培土TM-1、タキイ種苗(株))を充填したLeonard jar(Soil Science and Plant Nutrition,1983,29:97-100)に底面から一晩給水させたのち、ダイズ(品種「フクユタカ」、日光種苗(株))を1粒ずつ、表面から約1cm以内の深さに埋め込んで播種した。別途、YM(Yeast Extract Mannitol)培地(K2HPO4 0.5g、MgSO4・7H2O 0.2g、NaCl 0.1g、Yeast Extract 0.4g、Mannitol 10g、蒸留水1L(pH6.8))に1.5%の寒天(和光純薬工業(株))を加えて調製した寒天培地で根粒菌Bradyrhizobium japonicumNBRC14783T株を生育させ、これを容積50mLの試験管に調製した5mLのYM培地に一白金耳植菌し、30℃、250rpmにて24時間振盪培養した。その後、この根粒菌培養液を容積500mLの坂口フラスコに調製したYM培地100mLに1mL植菌し、30℃、120rpmにて振とう培養し、OD600が0.3程度まで菌を増殖させた。得られた根粒菌培養液1mLを、上記の播種した種子の上から滴下接種した。サポニン製剤(「大豆サポニン80」;アクセスワン(株))とカテキン製剤(「カテキン混合物,緑茶由来」;和光純薬工業(株)」;カテキン含量80%)について、それぞれ100ppmの水溶液、および各々の終濃度が100ppmとなるように調製したサポニン-カテキン混合水溶液を調製し、上記の播種した種子の上から200μL滴下した(「サポニン区」、「カテキン区」および「併用区」;該培土中、サポニン区および併用区のサポニン製剤の濃度はいずれも約0.16ppm、かつ製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約0.076ppmに相当)。対照として根粒菌培養液のみを滴下したものを準備した(「非施用区」)。
(ソヤサポニンBb含有大豆サポニン製剤およびリン酸鉄の併用がダイズ収量に与える効果)
ダイズ種子は「緑碧」(カネコ種苗(株))を使用した。5000分の1アールのWagnerポットに培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約4Lを充填し、培土表面から約1cmの深さに種子を3粒ずつ播種した。サポニン製剤(「大豆サポニン80」;(株)アクセスワン)50mg、リン酸鉄(III)四水和物(純正化学(株))0.75mg、およびそれらの混合物をそれぞれ、100mLの水道水に懸濁して培土表面に灌注した(それぞれ、「サポニン区」、「リン酸鉄区」および「併用区」;該培土中、併用区およびサポニン区におけるサポニン製剤の濃度は約12.5ppm、かつ製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約6.1ppmに相当)。対照として水道水100mLのみを同様に灌注した(「非施用区」)。播種した種子の上から、実施例4と同様に調製した根粒菌(根粒菌Bradyrhizobium japonicum NBRC14783T株)の菌液1mLを、マイクロピペッターを用いて滴下した。出芽後、ダイズ幼苗をポット当たり1株になるよう間引いた。光条件は播種から21日目までは明期16時間/暗期8時間、22日目以降は明期12時間/暗期12時間に設定した。温度は明期26℃/暗期20℃、湿度は50%とした。水の補給は、2~4日に1回、水道水を適量灌注することで行った。播種から86日後に莢(莢には子実を含む。以下同じ)を収穫し、株(ポット)当たりの莢新鮮重および莢当たり莢新鮮重(莢1個当たりの平均重量)を測定した(併用区のみn=7、それ以外はn=8)。処理区間の多重検定にはWilliams検定を用いた。
(ソヤサポニンBb含有大豆サポニン製剤およびリン酸鉄の併用が非耕作地土壌栽培ダイズの収量に与える効果)
ダイズ種子は「緑碧」(カネコ種苗(株))を使用した。5000分の1アールのWagnerポットに土壌(花王(株)栃木事業場構内から採取した、施肥等を行っていない非耕作地の土壌)約4Lを充填し、培土表面から約1cmの深さに種子を3粒ずつ播種した。サポニン製剤(「大豆サポニン80」;(株)アクセスワン)50mgおよびリン酸鉄(III)四水和物(純正化学(株))の混合物を100mLの水道水に懸濁して土壌表面に灌注した(「併用区」;該土壌中のサポニン製剤の濃度は約12.5ppm、かつ製造例1での定量結果(表7)から換算したグループB群ソヤサポニンの濃度として約6.1ppmに相当)。対照として水道水100mLのみを同様に灌注した(「非施用区」)。光条件は明期16時間/暗期8時間に設定した。温度は明期26℃/暗期20℃、湿度は50%とした。水の補給は、2~4日に1回、水道水を適量灌注することで行った。播種から83日後に莢(莢には子実を含む。以下同じ)を収穫して100℃で一晩乾燥させ、ポット当たりの莢乾燥重および子実のみの乾燥重を測定した。処理区ごとに最大値および最小値は除外して測定項目の平均値を算出した(併用区はn=5、非施用区はn=4)。処理区間の有意差検定にはStudentのt検定を用いた。
(ダイズ以外のマメ科植物に対するサポニン製剤の効果)
インゲンマメ(品種「軟莢つるなし」;日光種苗(株)およびエンドウ(品種「赤花つるなし絹莢豌豆」;日光種苗(株)より購入)の種子を、培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約4Lを充填した育苗箱(345mm×270mm×高さ75mm)に、培土表面から約1cmの深さにそれぞれ播種した。播種から8日後に、それぞれの幼植物体を培土から抜いて根を流水で洗浄し、所定の5倍の濃度に希釈したHoagland養液(Hoagland Modified Basal Salt Mixture(PhytoTechnology Laboratories社)を用いて調製)50mLを充填した50mL容のスクリュー管に1個体ずつ移した。該スクリュー管中のHoagland養液には、サポニン製剤(「大豆サポニン80」;(株)アクセスワン)を終濃度で0ppm(「非施用区」)、10ppm(「10ppm施用区」)または100ppm(「100ppm施用区」)添加した(該養液中のグループB群ソヤサポニンの濃度は、製造例1での定量結果(表7)からの換算濃度として0ppm、約4.9ppmおよび約49.1ppmに相当)。各施用区についてインゲンマメおよびエンドウそれぞれを5検体ずつ準備した。植物の栽培は人工気象器内で行い、光条件は明期16時間/暗期8時間、温度は明期26℃/暗期20℃、湿度は50%に設定した。養液は7日おきにサポニン製剤を含まない新しいものと入れ替えた。養液に移行してから14日後に、地上部新鮮重と地下部新鮮重の測定を行った。処理区間の多重検定にはDunnett検定を用いた。
(サポニン製剤の葉面散布によるダイズおよびヒヨコマメの生育促進効果)
培土(タキイ含水セル培土中期肥効型:バーミキュライト=1:1(体積比))約4Lを充填した育苗箱(タテ270mm×ヨコ345mm×高さ75mm)に、ダイズ(品種「フクユタカ」;日光種苗(株)より購入)およびヒヨコマメ(品種不明(カブリ型);日光種苗(株)より購入)の種子を培土表面から約1cmの深さにそれぞれ播種した。播種の際、ダイズの種子については約1cmの等間隔でタテ5個×ヨコ9個の配置となるようにし、ヒヨコマメの種子については約1cmの等間隔でタテ4個×ヨコ9個の配置となるようにした。育苗箱はバット上に設置し、当該バットに適宜水道水を補給することで給水を行った。栽培は人工気象器内で行い、光条件は明期16時間/暗期8時間、温度は明期26℃/暗期20℃、湿度は50%に設定した。ダイズについては、タテ5個×ヨコ3個の植物体群を1単位とした。ヒヨコマメについては、タテ4個×ヨコ3個の植物体群を1単位とした。両植物種のそれぞれ3単位を下記の葉面散布処理に供した(出芽しなかった個体があるため、実際のサンプル数としては後述の通りn=10~15の範囲となった)。
1)サポニン製剤の精製
大豆サポニン製剤(「大豆サポニン80」;(株)アクセスワン)4.99gを40v/v%エタノール300mLに溶解し、遠心して上清を回収した。合成吸着剤HP-20(三菱ケミカル)500mLをガラスカラムに充填し、エタノールで活性化し、40v/v%エタノールで平衡化したカラムに上清をアプライし、1000mLの40v/v%エタノール、続いて1000mLの60v/v%エタノール、最後に1000mLの99.5v/v%エタノールで溶出させた。各溶出液は減圧濃縮し、次いで凍結乾燥した。60v/v%エタノール溶出画分に比較的多くのグループB群のソヤサポニンが移行したことから、次に60v/v%エタノール画分からグループB群のソヤサポニンの精製を行った。凍結乾燥した60v/v%エタノール画分0.87gを60v/v%エタノール200mLに再溶解させ、そこへ活性炭(白鷺P、大阪ガスケミカル)1.0gを添加し、スターラーで1時間撹拌した。PTFEフィルターでろ過し、ろ液を減圧濃縮した後、さらに凍結乾燥して粉体の精製サポニン製剤を得た。大豆サポニン製剤および得られた精製サポニン製剤におけるグループB群のソヤサポニンの含有量を、以下の手順により定量した。
<LC-MS分析条件>
HPLC装置および質量分析装置は、それぞれShimadzu Nexera UHPLCシステム(島津株式会社)およびTripleQuad4500システム(株式会社エービー・サイエックス)を使用した。カラムはCapcell Core C18(2.1×50mm、2.7μm)およびガードカラムCapcell Core C18(2.1×5mm、2.7μm)(株式会社資生堂)を使用した。溶離液は、A:0.1v/v%ギ酸水、B:アセトニトリルを用い、グラジエント条件を0分~1分(10v/v%B)→1分~7分(1v/v%B~47.5v/v%B)→7分~9分(47.5v/v%B~85v/v%B)→9分~9.01分(85v/v%B~100v/v%B)→9.01分~10分(100v/v%B)→10分~10.01分(100v/v%B~10v/v%B)→10.01分~11分(10v/v%B)とした。流速は0.5mL/分とした。検出方法にはMRM法(多重反応モニタリング)を用い、極性はポジティブモードでおこなった。
標準品(製品番号、入手先)として、グループB群のソヤサポニンとしてソヤサポニンI(P2505、株式会社常磐植物化学研究所)、II(NP-000100、AnalytiCon Discovery)およびV(P2506、株式会社常磐植物化学研究所)を用い、それぞれについて検量線を作成した。
<サンプル>
実施例3で用いた大豆サポニン製剤(「サポニン,大豆由来」;和光純薬工業(株)) 実施例4で用いた大豆サポニン製剤(「大豆サポニン80」;(株)アクセスワン)
上記1)で得た精製サポニン製剤
各サンプルに含まれるソヤサポニンI、IIおよびVを検量線から定量した。定量値から算出した各製剤中のグループB群のソヤサポニンの総量(質量%)を表7に示す。
特開2017-190448号公報の記載に準じて、下記工程1および2により、土壌団粒化剤となるアルカリ処理リグニン(リグニン分解物)を製造した。
<工程1>
草本系バイオマスとして、サトウキビバガスを、乾燥質量として30gガラス瓶に入れ、固形分含有量が10質量%になるように、1.6質量%水酸化ナトリウム水溶液を加えた。ガラス瓶をオートクレーブで、95℃、6時間加熱して反応物を得た。
工程1で得られた反応物を、400メッシュのSUSメッシュとヌッチェを用いて減圧濾過した。残渣を、90℃のイオン交換水300mLで洗浄した。ろ液と洗浄液を集め、1.0M塩酸でpH4にしてリグニン分解物を含む懸濁液を得た。
Claims (24)
- ソヤサポゲノールBの配糖体であって、該ソヤサポゲノールBのC-22位がヒドロキシ基であり、かつ該ソヤサポゲノールBのC-3位ヒドロキシ基に糖が結合している配糖体を有効成分とするマメ科植物生育促進剤。
- 前記ソヤサポゲノールBの配糖体がソヤサポニンBbである、請求項1記載のマメ科植物生育促進剤。
- 前記マメ科植物が、ダイズ属植物、インゲンマメ属植物、ヒヨコマメ属植物、エンドウ属植物、ヒラマメ属植物、キマメ属植物、ソラマメ属植物、およびラッカセイ属植物からなる群から選択される少なくとも1種である、請求項1又は2記載のマメ科植物生育促進剤。
- 前記マメ科植物が、ダイズ、インゲンマメ、ヒヨコマメ、エンドウ、レンズマメ、キマメ、ソラマメおよびラッカセイからなる群から選択される少なくとも1種である、請求項3記載のマメ科植物生育促進剤。
- マメ科植物用の栽培基材、農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントである、請求項1~4のいずれか1項記載のマメ科植物生育促進剤。
- ソヤサポゲノールBの配糖体であって、該ソヤサポゲノールBのC-22位がヒドロキシ基であり、かつ該ソヤサポゲノールBのC-3位ヒドロキシ基に糖が結合している配糖体を有効成分として用いるマメ科植物の生育促進方法。
- 前記マメ科植物を前記ソヤサポゲノールBの配糖体とともに栽培することを含む、請求項6記載の方法。
- 前記ソヤサポゲノールBの配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントを栽培基材へ添加すること、および、
得られた該配糖体を添加した栽培基材で前記マメ科植物を栽培すること、
を含む、請求項7記載の方法。 - 前記ソヤサポゲノールBの配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントを播種前のマメ科植物種子へ塗布または塗抹すること、および、
該種子を栽培基材で栽培すること、
を含む、請求項7記載の方法。 - 前記ソヤサポゲノールBの配糖体を有効成分として含有する農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントをマメ科植物の植物体へ散布、噴霧または塗布すること、および、
該植物体を栽培基材で栽培すること、
を含む、請求項7記載の方法。 - 前記農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントが前記ソヤサポゲノールBの配糖体を0.0005~80質量%含有する、請求項8~10のいずれか1項記載の方法。
- 前記栽培の際に、前記栽培基材中における前記ソヤサポゲノールBの配糖体の濃度が0.01~100質量ppmである、請求項7~10のいずれか1項記載の方法。
- 前記ソヤサポゲノールBの配糖体がソヤサポニンBbである、請求項6~12のいずれか1項記載の方法。
- 前記マメ科植物が、ダイズ属植物、インゲンマメ属植物、ヒヨコマメ属植物、エンドウ属植物、ヒラマメ属植物、キマメ属植物、ソラマメ属植物、およびラッカセイ属植物からなる群から選択される少なくとも1種である、請求項6~13のいずれか1項記載の方法。
- 前記マメ科植物が、ダイズ、インゲンマメ、ヒヨコマメ、エンドウ、レンズマメ、キマメ、ソラマメおよびラッカセイからなる群から選択される少なくとも1種である、請求項14記載の方法。
- 栽培基材として、アルカリ処理リグニンを用いて団粒化処理した土壌もしくは培土を用いる、請求項6~15のいずれか1項記載の方法。
- 前記ソヤサポゲノールBの配糖体がカテキン類またはリン酸鉄(III)と併用される、請求項6~16のいずれか1項記載の方法。
- ソヤサポゲノールBの配糖体であって、該ソヤサポゲノールBのC-22位がヒドロキシ基であり、かつ該ソヤサポゲノールBのC-3位ヒドロキシ基に糖が結合している配糖体の、マメ科植物生育促進剤の製造のための使用。
- 前記マメ科植物生育促進剤が農薬、肥料、微生物資材、土壌改良剤、播種用資材または植物用サプリメントであり、前記ソヤサポゲノールBの配糖体を0.0005~80質量ppm含有する、請求項18記載の使用。
- 前記マメ科植物生育促進剤が栽培基材であり、前記ソヤサポゲノールBの配糖体を0.01~100質量ppm含有する、請求項18記載の使用。
- 前記ソヤサポゲノールBの配糖体がソヤサポニンBbである、請求項18~20のいずれか1項記載の使用。
- 前記マメ科植物が、ダイズ属植物、インゲンマメ属植物、ヒヨコマメ属植物、エンドウ属植物、ヒラマメ属植物、キマメ属植物、ソラマメ属植物、およびラッカセイ属植物からなる群から選択される少なくとも1種である、請求項18~21のいずれか1項記載の使用。
- 前記マメ科植物が、ダイズ、インゲンマメ、ヒヨコマメ、エンドウ、レンズマメ、キマメ、ソラマメおよびラッカセイからなる群から選択される少なくとも1種である、請求項22記載の使用。
- 前記マメ科植物生育促進剤がカテキン類またはリン酸鉄(III)をさらに含有する、請求項18~23のいずれか1項記載の使用。
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US11632960B2 (en) | 2017-10-17 | 2023-04-25 | Kao Corporation | Plant growth promoter |
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