Plant growth–promoting composition

Abstract

A plant growth–promoting composition containing, as an active ingredient, one or more selected from the group consisting of grape extracts and grape powders.

Description

Plant growth promoting composition

[0001] The present invention relates to a plant growth promoting composition and a method for promoting plant growth.

[0002] With the current explosive growth in the human population, producing a sufficient supply of plants for food has become a global challenge. Furthermore, desertification due to the reduction in green space has become a problem, and rising earth temperatures due to global warming have also become a problem, resulting in an increasing number of environmental problems affecting plant growth. In other words, excessive stress on plants caused by global environmental factors often causes problems for plant growth.

[0003] Therefore, there is a demand for technologies that promise more efficient harvesting than conventional technologies, and biostimulants are attracting attention as a new technology that reduces damage to plants caused by climate and soil conditions by controlling environmental stress on plants, thereby providing healthy plants.

[0004] Patent Document 1 discloses a plant stomatal opening promoter containing stevia, and reports that artificially promoting plant stomatal opening with stevia improves heat stress tolerance. Patent Document 2 discloses a composition for activating a gene that activates the ethylene and / or jasmonic acid signaling system, containing a yeast cell wall decomposition product, and reports that applying the yeast cell wall decomposition product to a plant activates the expression of a plant defensin gene involved in the antibacterial properties and disease prevention of the plant. Patent Document 3 discloses a method for increasing grain yields by inoculating a plant for cultivation with a substance containing the cell wall of a methanol-assimilating bacterium.

[0005] Patent No. 6741262 Patent No. 4931388 JP2019-180361

[0006] As mentioned above, research has been conducted on improving environmental stress tolerance and increasing yield in plants using various materials, but no examples have been reported using grape extract or grape powder.

[0007] As a result of extensive research, the inventors have discovered that, by using a highly accurate screening method that they developed themselves, grape extract or grape powder, among various materials, is an effective ingredient for promoting plant growth, and have thus completed the present invention.

[0008] That is, the present invention is as follows: [1] A plant growth-promoting composition containing one or more active ingredients selected from the group consisting of grape extract and grape powder. [2] The plant growth-promoting composition according to [1] above, which has one or more plant growth-promoting effects selected from the group consisting of stem, leaf, or root elongation, increased leaf number, promotion of flowering or fruiting, increased flower or fruit number, increased plant weight or crop yield, greening, and promotion of tillering. [3] The plant growth-promoting composition according to [1] or [2] above, which has an effect of improving environmental stress tolerance. [4] The plant growth-promoting composition according to [3] above, which has an effect of improving environmental stress tolerance to one or more stresses selected from the group consisting of temperature stress, nutritional stress, chemical stress, light stress, drought stress, pH stress, salt stress, hypoxic stress, herbicide stress, physical stress, and disease stress. [5] The plant growth-promoting composition according to [3] or [4] above, which activates a gene in a stress response system. [6] A method for promoting plant growth, comprising applying a plant growth-promoting composition containing one or more active ingredients selected from the group consisting of grape extract and grape powder to a plant, soil in which the plant grows, or a culture solution. [7] The method for promoting plant growth according to [6], wherein the amount of the plant growth-promoting composition used is 0.001 to 50% by mass or 0.001 to 50% by volume relative to the culture solution or dilution solvent.

[0009] By applying a composition containing one or more selected from the group consisting of grape extract and grape powder of the present invention to plants, plant growth can be promoted more safely than with conventional materials.

[0010] 1A and 1B are images showing the improvement in root establishment of rice plants without the addition of grape extract or grape powder (left) and with the addition of 0.45 g of a plant growth-promoting composition containing grape powder (Example 1) (right). 1C and 1D are images showing the improvement in root establishment of rice plants without the addition of grape extract or grape powder (left) and with the addition of 2.7 mL of a plant growth-promoting composition containing grape extract (Example 2) (right). 1D and 1D are images showing the improvement in root establishment of rice plants without the addition of grape extract or grape powder (A) and with the addition of 2.7 mL of a plant growth-promoting composition containing grape extract (B).

[0011] Hereinafter, a mode for carrying out the present invention (hereinafter also referred to as "the present embodiment") will be described in detail. Note that the present invention is not limited to the present embodiment, and various modifications can be made within the scope of the gist of the present invention.

[0012] <Plant Growth-Promoting Composition> The plant growth-promoting composition of this embodiment contains one or more active ingredients selected from the group consisting of grape extract and grape powder. Grape (Vitis vinifera) is a plant of the genus Vitis in the family Vitaceae. Specific examples include Vitis vinifera, and more specifically Vitis labrusca. The grape variety is not particularly limited, and may be a variety for eating fresh, a variety for processed products such as wine, a variety domestic or foreign to Japan, or a black grape variety, red grape variety, or white grape variety. Examples of grape varieties include Kyoho, Fujiminori, Pione, Muscat Bailey A, Delaware, Ruby Roman, Queen Nina, Kaiji, Koshu, Shine Muscat, Muscat of Alexandria, Niagara, Cabernet Sauvignon, Merlot, Syrah, Zinfandel, Campbell Early, Zweigelt, Rondo, Kerner, Muscat, Muller-Thurgau, Pinot Noir, Chardonnay, Riesling, Sauvignon Blanc, Nagano Purple, Rosario Bianco, King Dela, Honey Venus, Beni Ballad, Goldfinger, Suiho, Wink, Gorby, Aki Queen, Manicure Finger, Oriental Star, Shinano Smile, Akamine, New Pione, Aurora Black, Shion, etc. The origin of the grapes is also not particularly limited.

[0013] In this embodiment, the grape extract is an extract of specific components from various parts of grapes, and is not particularly limited. The grape extract can be obtained, for example, by the method described below in "Method for preparing a plant growth-promoting composition." Furthermore, the grape powder is obtained by drying and pulverizing various parts of grapes or grape extract, and is not particularly limited.

[0014] The plant growth-promoting composition in this embodiment is a composition that has a plant growth-promoting effect, and here, the plant growth-promoting effect includes, for example, growth-promoting effects such as elongation of stems, leaves, or roots, an increase in the number of leaves, promotion of flowering or fruiting, an increase in the number of flowers or fruits, an increase in plant weight or crop yield, greening, or promotion of tillering.

[0015] More specifically, a plant growth-promoting effect refers to an increase in actual weight gain (%) or dry weight gain (%) when the plant growth-promoting composition of the present embodiment is used compared to when it is not used. The actual weight or dry weight may be the actual weight or dry weight of the entire plant, or the actual weight or dry weight of each part of the plant (preferably the root). When the plant growth-promoting composition is used, the actual weight gain (%) or dry weight gain (%) is preferably increased by 10% or more, more preferably 20%, and even more preferably 30% or more compared to when the plant growth-promoting composition of the present embodiment is not used. A plant growth-promoting effect includes a case where no growth is observed without the use of the plant growth-promoting composition, but growth is observed after use of the plant growth-promoting composition. Note that the above values ​​are average values ​​for the entire plant to which the plant growth-promoting composition is applied.

[0016] In addition, the plant growth-promoting effect also includes the effect of improving the activity of plants, such as: 1) root elongation and improved root survival rate; 2) increased nutrient absorption efficiency; 3) flower bud induction, promotion of flowering, and increased fruit production; 4) accumulation of sugars and other components; 5) increased water-saving effect; 6) callus induction effect; 7) increased plant size, particularly elongation or thickening of above-ground and below-ground parts; and 8) wound repair. In other words, the plant growth-promoting composition of this embodiment may have the effects 1) to 8) above in addition to growth-promoting effects such as elongation of stems, leaves, or roots, increased leaf number, promotion of flowering or fruiting, increased number of flowers or fruits, increased plant weight or crop yield, greening, or promotion of tillering. The exertion of effect 1) above may promote nutrient absorption by the roots, thereby promoting the growth of the entire plant.

[0017] The plant growth-promoting composition of this embodiment may confer tolerance to a variety of environmental stresses to which a plant is exposed. The plant growth-promoting composition of this embodiment may have the effect of improving environmental stress tolerance. Here, examples of environmental stress include biotic stress and abiotic stress, and specific examples include herbicide stress, disease stress, temperature stress such as high temperature stress and low temperature stress, nutritional stress, chemical stress, light stress, drought stress, pH stress such as oxidative stress, salt stress such as osmotic stress, hypoxic stress, and physical stress. Nutritional stress includes, for example, nutrient deficiency stress and nutrient excess stress.

[0018] In this embodiment, "environmental stress tolerance" refers to a trait that allows a plant to grow normally or near-normally even under growth conditions that cause environmental stress, without substantially suffering from undesirable effects such as inability to grow (withering), poor growth (e.g., bleaching or yellowing of the plant, reduced root length or reduced leaf number), reduced growth rate, or reduced plant weight or crop yield. By improving the plant's environmental stress tolerance with the plant growth-promoting composition of this embodiment, the plant can grow normally even in external environments that cause undesirable effects on the growth of ordinary plants.

[0019] The temperature stress tolerance-imparting effect of the plant growth-promoting composition of this embodiment includes, for example, the effect of promoting normal growth under high-temperature and low-temperature conditions. High-temperature conditions may be appropriately determined by a person skilled in the art based on common general knowledge in the technical field, but are preferably 35°C or higher, for example, 35°C to 50°C. Low-temperature conditions may be appropriately determined by a person skilled in the art based on common general knowledge in the technical field, but are preferably 10°C or lower, for example, 0°C to 10°C. Furthermore, the salt stress-imparting effect enables normal growth in the presence of 0.1 to 3.0% by mass of sodium chloride. The drought stress-imparting effect enables normal growth even at a relative humidity of 10 to 40%. The nutritional stress-imparting effect of the plant growth-promoting composition of this embodiment includes, for example, the effect of promoting normal growth in plants that are deficient in chemical substances that serve as nutrient sources for plants (nutrient deficiency). Here, the nutrient deficiency state may be, for example, a state in which the amount of each nutrient is less than the concentration indicated in the test period in Table 1 of the Examples, and specifically, a state in which the amount of each nutrient is about one-half to one-third, one-third to one-quarter, one-quarter to one-fifth, one-fifth to one-sixth, one-sixth to one-seventh, or one-seventh to one-eighth of the amount indicated in the test period concentration in Table 1. The test period concentrations in Table 1 are concentrations generally used in the cultivation of crops such as rice.

[0020] In this embodiment, the environmental stress tolerance of a plant can be evaluated, for example, by the following means. The presence or absence of salt stress tolerance can be confirmed by growing a target plant in a medium or soil to which an appropriate concentration of salt has been added under conditions of a growth temperature and growth period that are suitable for the plant, and evaluating the phenotype. The presence or absence of drought stress tolerance can be confirmed by growing a target plant under conditions of a growth temperature, growth humidity, and growth period that are dry for the plant, and evaluating the phenotype. Furthermore, the presence or absence of high temperature stress tolerance can be confirmed by acclimating a target plant to a high temperature, such as 35°C to 50°C, and then growing it under conditions of a growth temperature and growth period that are suitable for the plant, and evaluating the phenotype.

[0021] When the plant growth-promoting composition of this embodiment has an effect of improving environmental stress tolerance, it may activate a gene in a stress response system. Examples of stress response genes include, but are not limited to, genes generally known as indicators of response to environmental stresses such as temperature stress, nutritional stress, chemical stress, light stress, drought stress, pH stress, salt stress, hypoxic stress, herbicide stress, physical stress, and disease stress. Genes generally known as indicators of response to environmental stresses include genes disclosed in publicly known databases such as the National Center for Biotechnology Information (NCBI), Ensemble Plants, The Arabidopsis Information Resource (TAIR), and The Rice Annotation Project (RAP). Specifically, genes generally known as indicators of a response to high temperature stress include Os06g0682900, Os11g0216100, Os03g0426900, Os08g0127600, Os03g0161900, Os05g0460000, Os05g0530400, etc. For example, by applying the plant growth-promoting composition of this embodiment to a plant, the expression levels of genes such as Os06g0682900, Os11g0216100, Os03g0426900, Os08g0127600, Os03g0161900, Os05g0460000, and Os05g0530400 in the plant acclimated to high temperature are increased compared to the control.

[0022] The plant growth-promoting composition of this embodiment may contain one or more selected from the group consisting of grape extract and grape powder, either alone or in combination with one or more agriculturally acceptable ingredients. Examples of agriculturally acceptable ingredients include, but are not limited to, solvents, carriers, excipients, binders, solubilizers, stabilizers, thickeners, leavening agents, lubricants, surfactants, oily liquids, buffers, disinfectants, antifreeze agents, antifoaming agents, colorants, antioxidants, preservatives, and additional active ingredients. Preferred agriculturally acceptable carriers include water, mineral oil fractions such as kerosene or diesel oil, vegetable or animal oils, cyclic or aromatic hydrocarbons (e.g., paraffin, tetrahydronaphthalene, alkylated naphthalenes or their derivatives, or alkylated benzenes or their derivatives), alcohols (e.g., methanol, ethanol, propanol, butanol, or cyclohexanol), ketones (e.g., cyclohexanone), amines (e.g., N-methylpyrrolidone), and mixtures thereof.

[0023] When the plant growth-promoting composition of this embodiment contains one or more additional active ingredients, various compounds known in the art having an activity of improving environmental stress tolerance can be used as the additional active ingredients. When the plant growth-promoting composition contains such one or more additional active ingredients, the environmental stress tolerance of the plant tends to be further improved.

[0024] The plant growth-promoting composition of this embodiment can be used in conjunction with conventionally known agricultural materials, and examples of such agricultural materials include pesticides, fertilizers, soil conditioners, and potting soil (culture soil). Specifically, the pesticides may be fungicides, insecticides, insecticides, fungicides, miticides, nematicides, rodenticides, insect repellents, attractants, spreaders, hormones (growth regulators), herbicides, and the like; the fertilizers may be organic fertilizers, chemical fertilizers, liquid fertilizers, solid fertilizers, compound fertilizers, compost, growth promoters, stimulants, calcium supplements, and the like; the soil conditioners may be those intended for physical, chemical, or biological soil improvement, and may also be oxygen suppliers, and the like. In addition, the pesticides, fertilizers, soil conditioners, and potting soil may contain components derived from plants, animals, lime, ores, or microorganisms.

[0025] The content of one or more selected from the group consisting of grape extract and grape powder in the plant growth-promoting composition relative to the entire plant growth-promoting composition may be any value as long as the one or more selected from the group consisting of grape extract and grape powder functions as an active ingredient for promoting plant growth, but may be, for example, 5, 10, 20, 30, 40, 50, 60, 70, 80, or 90% by mass (volume %) or more, and may be 99, 95, 90, 80, 70, 60, 50, 40, 30, 20, or 10% by mass (volume %) or less.

[0026] The plant growth-promoting composition of this embodiment can be used as an agricultural chemical formulation or pesticide to promote plant growth. The plant growth-promoting composition may be in any form, such as a solid (e.g., powder or granules) or a liquid (e.g., a solution or suspension), and is preferably used in the form of a liquid such as a solution or suspension. When used as a solution, it may be in a liquid state, or may be used as a liquid prepared just before use when applied.

[0027] The formulation of the plant growth-promoting composition of this embodiment is not particularly limited, and can be formulated into formulations commonly used in the art, such as emulsions, wettable powders, liquids, water-soluble powders, dusts, powders, pastes, or granules.

[0028] In this embodiment, the plant to which the plant growth-promoting composition is applied is not particularly limited, and examples thereof include angiosperms and gymnosperms. Examples of target plants include Asteraceae plants such as chrysanthemums and gerberas, Solanaceae plants such as potatoes, tomatoes, and eggplants, Brassicaceae plants such as rapeseed and rapeseed, Poaceae plants such as rice, corn, wheat, sugarcane, and barley, Legumes such as soybeans, Apiaceae plants such as carrots, Lamiaceae plants such as basil, mint, and rosemary, Convolvulaceae plants such as morning glory, Salicaceae plants such as poplars, and Euphorbiaceae plants such as castor beans, cassava, and jatropha. Convolvulaceae plants such as sweet potato, Rutaceae plants such as oranges and lemons, Rosaceae plants such as cherry blossoms and roses, Orchidaceae plants such as moth orchids, Gentianaceae plants such as bellflowers, Primulaces such as cyclamen, Violets such as pansies, Liliaceae plants such as lilies, Amaranthaceae plants such as sugar beets, Vitaceae plants such as grapes, Cupressaceae plants such as cedars and cypresses, Oleaceae plants such as olives and Osmanthus, and Pinaceae plants such as red pine.

[0029] The target plant may be not only the whole plant (i.e., the complete plant), but also parts of the plant, such as plant tissues or organs (e.g., cut flowers or vegetative propagation organs such as rhizomes, tubers, corms or runners), cultured cells and / or callus.

[0030] The plant growth-promoting composition of this embodiment can be applied to the whole or part of a plant at any stage of growth, including before or after plant emergence (e.g., the whole or part of a seed, seedling, or mature plant).

[0031] <Method for Preparing Plant Growth-Promoting Composition> A plant growth-promoting composition can be prepared, for example, by obtaining one or more selected from the group consisting of grape extract and grape powder, and then mixing them with the agriculturally acceptable ingredients described above, as needed. The method for obtaining grape extract is not particularly limited, and a general method for extracting plant extracts can be used. Specifically, the composition can be obtained by a method including the steps of: heating or crushing one or more selected from the group consisting of grape leaves, stems, skins, stalks, seeds, and fruits, and then removing impurities to obtain a grape extract raw material; concentrating the grape extract raw material obtained in the above step to obtain a grape concentrate; and fractionating the grape concentrate obtained in the above step to obtain a grape extract containing specific components. Alternatively, grape extract can be obtained using one or more selected from the group consisting of grape leaves, stems, skins, stalks, seeds, and fruits that have been subjected to the respective steps for obtaining grape extract as a raw material. For example, a grape extract raw material or a grape concentrate can be obtained from grape residue, such as impurities removed in the process of obtaining a grape extract raw material, using a method similar to that described above, and then grape extract can be obtained from the grape extract raw material or grape concentrate using a method similar to that described above. Alternatively, grape extract can be obtained using grape residue, such as impurities obtained during the production process of any grape processed product, as a raw material. The grape extract raw material or grape concentrate can be used as the grape extract as is. During or after each process, the grape extract raw material, grape concentrate, or grape extract can be heated or pressurized. When performing the heat treatment or pressurization treatment, a solvent can be added as appropriate. The conditions for obtaining the grape extract can be determined appropriately depending on the variety, part, condition, etc. of the grapes used. The method for obtaining grape powder is not particularly limited, and the grape powder can be obtained by drying fresh or frozen grape residue such as impurities generated when obtaining grape extract raw materials, grape concentrates, or grape extract, grape extract raw materials, grape concentrates, grape extract, or grapes (one or more selected from the group consisting of grape leaves, stems, skins, stalks, seeds, and fruits) by a method such as hot air drying, air drying, dry heat drying, or vacuum drying, and then crushing the dried grapes into powder.

[0032] (Crushing step) Grapes can be crushed using fresh grapes or dried grapes, for example, using a mixer. The crushed grapes are filtered using a filter cloth, filter paper, or the like to obtain a grape extract raw material from which impurities have been removed. When filtering, an extraction solvent such as an organic solvent may be added in advance. Alternatively, instead of filtration, impurities may be removed by collecting the supernatant using a centrifuge.

[0033] (Concentration Step) The obtained grape extract raw material can be concentrated using a method such as vacuum concentration, freeze concentration, or membrane concentration to obtain a grape concentrate. The concentration ratio during concentration is preferably 3 to 30 times, and more preferably 4 to 20 times. A concentration ratio of 5 times or more tends to reduce the amount of plant growth-promoting composition added to plants, improving handling and reducing transportation costs. A concentration ratio of 30 times or less tends to reduce the amount of precipitate in the plant growth-promoting composition, making it easier to handle. Note that concentration of the grape extract raw material is not essential in preparing the plant growth-promoting composition; the obtained grape extract raw material may be directly subjected to the fractionation step described below.

[0034] (Fractionation step) The obtained grape concentrate can be fractionated by solvent extraction or the like to obtain a grape extract containing specific components. In the fractionation step, for example, a hydrophilic fraction and a hydrophobic fraction can be obtained by liquid-liquid partitioning. Further, by passing the fraction through a column, a more purified fraction can be obtained.

[0035] The grape extract obtained in the fractionation step may be further purified or subjected to a separation process for a highly active fraction, if necessary. Examples of the purification process include filtration, adsorption (ion exchange resin column, activated carbon column, etc.), etc. Examples of the separation process for a highly active fraction include gel filtration, adsorption, silica gel column chromatography, HPLC, etc.

[0036] <Plant Growth Promotion Method> The plant growth promotion method of this embodiment is a method for promoting plant growth by applying a composition containing one or more active ingredients selected from the group consisting of the above-mentioned grape extract and grape powder to a plant, or to the soil or culture solution in which the plant grows. In this method, the plant growth promotion composition can be applied to a plant or a part thereof (e.g., a seed, a seedling, or a mature plant) at any growth stage, including before or after germination. Furthermore, the composition can be applied not only to the plant itself, but also to the soil, culture medium, or culture solution in which the plant grows. Plant growth can be promoted by applying the plant growth promotion composition to a plant at a growth stage or to the soil, culture medium, or culture solution in which the plant grows.

[0037] The plant growth-promoting composition may be applied to a plant once or multiple times. Furthermore, when the plant growth-promoting composition is applied at multiple application periods, the plant growth-promoting composition may be applied once or multiple times at each application period.

[0038] Examples of means for applying the plant growth-promoting composition to plants include addition, spraying, coating, and immersion. These may be performed alone or in combination of two or more. The plant growth-promoting composition may be applied to plants manually or by machine. When using a machine, the method is not particularly limited, but application (spraying) may be performed using, for example, a drone.

[0039] The amount of the plant growth-promoting composition applied to plants, when applied to a culture solution in which plants grow, or when diluted with a diluent solvent and applied to a field or the like in which plants grow, is preferably 0.001 to 50% by mass or 0.001 to 50% by volume, more preferably 0.01 to 30% by mass or 0.01 to 30% by volume, and even more preferably 0.01 to 10% by mass or 0.01 to 10% by volume, relative to the culture solution or diluent. When the amount of the plant growth-promoting composition used is within the above range, a sufficient plant growth-promoting effect tends to be obtained. However, the amount used may be adjusted appropriately depending on the content of one or more selected from the group consisting of grape extract and grape powder in the plant growth-promoting composition, the type and growth state of the plant to which the plant growth-promoting composition is applied, and the plant's growth environment, such as temperature, humidity, and light. The amount used may be determined by investigating a concentration range in which an effect is obtained using a known cultivation test method. When a plant growth-promoting composition diluted with a diluting solvent is applied to a plant using a drone, the amount of solution that can be carried on the drone is limited by the drone's carrying capacity, etc., and therefore the amount of plant growth-promoting composition used relative to the diluting solvent is preferably 1 to 50% by mass or 1 to 50% by volume.

[0040] The components of the culture medium and dilution solvent are not particularly limited as long as they do not adversely affect the growth of the plant, and may contain water, pesticides, etc.

[0041] The preferred range of the amount used varies depending on the concentration of the grape extract raw material, but the preferred range is when the concentration is 3 to 30 times.

[0042] The present invention will be explained in more detail using examples and comparative examples, but the present invention is not limited to these examples in any way.

[0043] (Method for measuring root weight) The roots of the rice plants were identified based on their color and shape, and the weight of the identified roots was measured in a bottle using a microbalance.

[0044] Example 1: Grape powder was obtained by drying and pulverizing grape skins cut at 3 cm intervals. Rice was grown in the seedling culture medium shown in Table 1. Rice was then transferred to 900 mL of the test culture medium shown in Table 1, and 0.45 g (0.05% by mass) of the grape powder obtained above was added. The effects on the rice were investigated. As a result, the weight of the rice roots increased by 25.1% compared to the case without grape extract, demonstrating an improved effect on root establishment (Figure 1). Note that the concentration of nutrients during the seedling stage was approximately one-sixth to one-seventh of the concentration during the test stage, which was sufficient for rice growth, indicating that the rice plants supplemented with grape extract were nutrient-deficient. Example 2: Grape extract raw material was obtained from 20 g of a mixture containing grape skins and twigs, and the grape extract raw material was then concentrated approximately five times to obtain grape extract. Rice was grown in the seedling culture medium shown in Table 1. The rice plants were then transferred to 900 mL of the culture medium for the test period shown in Table 1, and 2.7 mL (0.3% by volume) of the grape extract obtained above was added to investigate its effect on the rice. The results showed that the weight of the rice roots increased by 31.1% compared to when grape extract was not added, demonstrating an improved effect on the root development of rice (Figure 2). Furthermore, since the concentration of nutrients during the seedling stage was approximately one-sixth to one-seventh of the concentration during the test period, which is sufficient for rice growth, the rice plants to which grape extract was added were in a state of nutritional deficiency.

[0045]

[0046] Example 3 Rice seedlings grown in the same manner as in Example 2 were transferred to 900 mL of the culture medium for the test period shown in Table 1, and 2.7 mL (0.3% by volume) of the same grape extract as in Example 2 was added. After cultivation for several days in this state, the rice was cultivated under high-temperature conditions of 35°C or higher to investigate the effect of high-temperature conditions on the rice. As a result, when grape extract was not added (Rice A), leaf death and yellowing of the above-ground parts were evident, whereas when grape extract was added (Rice B), the progression of leaf death and yellowing was suppressed (Figure 3).

[0047] [Example 4] The changes in gene expression in rice A and B were also evaluated.

[0048] (A) Sample preparation Preparation of rice plants RNA was extracted from the tissues of rice plants A and B. RNA extraction RNA samples were obtained using a Maxwell RSC Plant RNA Kit (Promega) and a Maxwell RSC Instrument (Promega) according to the manufacturer's instructions. - RNA quality confirmation Measurement equipment: Agilent 5400 Bioanalyzer <5312AA> (Agilent Technologies), reagent kit: Agilent RNA kit (15nt) <DNF-471> (Agilent Technologies) and analysis software: ProSize (Agilent Technologies) were used to confirm that there were no problems with the quality of the RNA sample, and then used for library preparation. - Library preparation A library for the next-generation sequencer DNBSEQ T7 was prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina according to the following steps. 1. Purification of mRNA using oligo(dT) magnetic beads. 2. Fragmentation. 3. Synthesis of single-stranded and double-stranded cDNA using random primers. 4. End repair, polyadenylation, adapter binding, size selection, and amplification are performed.

[0049] (B) Analysis using a next-generation sequencer Sequence analysis Instrument: DNBSEQ T7 (MGI) Sequencing (overview) Sequencing was performed using the DNBSEQ T7 according to the following steps: 1. Addition of sequencing reagents 2. Single-base extension reaction 3. Removal of unreacted bases 4. Incorporation of fluorescent signal 5. Removal of protecting groups and fluorescence 6. Repeat cycles of 2 cycles... 3 cycles... up to 300 cycles

[0050] (C) Data Analysis - Primary Data Analysis Analysis computer OS: Ubuntu Desktop 22.04.4 LTS Analysis software: FastQC, hisat2, samtools, HTSeq, DESeq2 Analysis was performed using the above software. 1. Read Quality Verification FastQC was used to verify the quality of the bases contained in the obtained fastq file. 2. Mapping and Sorting Hisat2 was used to map to the Oryza sativa reference genome, Oryza sativa Japonica Group (assembly IRGSP-1.0). Subsequently, the files were sorted and converted to binary data using samtools. 3. Read Count The number of mapped reads was counted using htseq-count in HTSeq. 4. Statistical Analysis The gene expression levels in rice A and B were compared using the DESeq2 package in the statistical package R.

[0051] [Results] Comparison of expression levels between two groups A comparison between the two groups was performed using a generalized linear model. The fold change values ​​(= rice B / rice A) and p-values ​​of genes whose expression levels were significantly increased in the grape extract-treated group are shown in Table 2 below.

[0052]

Claims

1. A plant growth promoting composition comprising, as an active ingredient, one or more selected from the group consisting of grape extract and grape powder.

2. The plant growth-promoting composition according to claim 1, which has one or more plant growth-promoting effects selected from the group consisting of elongation of stems, leaves or roots, increased leaf number, promotion of flowering or fruiting, increased number of flowers or fruits, increased plant weight or crop yield, greening, and promotion of tillering.

3. A plant growth-promoting composition according to claim 1 or 2, which has the effect of improving environmental stress tolerance.

4. A plant growth-promoting composition as described in claim 3, which has the effect of improving environmental stress tolerance against one or more stresses selected from the group consisting of temperature stress, nutritional stress, chemical stress, light stress, drought stress, pH stress, salt stress, hypoxic stress, herbicide stress, physical stress, and disease stress.

5. The plant growth-promoting composition according to claim 3, which activates genes in the stress response system.

6. A method for promoting plant growth, comprising applying to a plant, or to soil or culture solution in which the plant grows, a plant growth-promoting composition containing one or more active ingredients selected from the group consisting of grape extract and grape powder.

7. The plant growth promoting method according to claim 6, wherein the amount of the plant growth promoting composition used is 0.001 to 50% by mass or 0.001 to 50% by volume relative to the culture solution or dilution solvent.

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