Composition for increasing yield of plants of leafy vegetables, fruit vegetables, and legumes

Abstract

Provided is a composition for increasing the yield of plants such as leafy vegetables, fruit vegetables, and legumes, said composition comprising a cell-wall-containing substance of methanol-assimilating bacteria.

Description

Composition for increasing yield of leafy vegetables, fruit vegetables and leguminous plants

[0001] The present invention relates to a composition for increasing the yield of leafy vegetables, fruit vegetables and leguminous plants, a method for increasing the yield of leafy vegetables, fruit vegetables and leguminous plants, and a method for cultivating leafy vegetables, fruit vegetables and leguminous plants.

[0002] Currently, with the explosive growth of global population, producing a sufficient supply of agricultural products has become a worldwide issue. Application of fertilizers is indispensable for improving productivity and achieving stable production of agricultural crops. However, from the perspectives of environmental burden and price instability caused by resource localization, technologies for reducing fertilizer application amount are in demand.

[0003] Therefore, in recent years, biostimulant materials that ensure crop yield and quality while reducing fertilizer application amount have attracted attention as a new technology.

[0004] Patent Document 1 discloses a technique for infecting rice with nitrogen-fixing bacteria in order to promote the growth of rice and increase seed yield.

[0005] Japanese Unexamined Patent Publication No. 2003-274779

[0006] As described above, although studies on increasing plant yield using nitrogen-fixing bacteria have been conducted so far, there has been no report on examples of using cell wall-containing substances of methanol-assimilating bacteria on leafy vegetables, fruit vegetables and leguminous plants.

[0007] As a result of intensive studies, the present inventors have found that the yield can be increased by inoculating leafy vegetables, fruit vegetables and leguminous plants with a composition containing cell walls of methanol-assimilating bacteria, and have completed the present invention.

[0008] In other words, the present invention is as follows: [1] A composition for increasing the yield of leafy vegetables, comprising a cell wall-containing substance of methanol-utilizing bacteria. [2] A composition for increasing the yield of fruit vegetables, comprising a cell wall-containing substance of methanol-utilizing bacteria. [3] A composition for increasing the yield of legumes, comprising a cell wall-containing substance of methanol-utilizing bacteria. [4] The yield-increasing composition according to any one of [1] to [3], wherein the methanol-utilizing bacteria is Methylobacterium sp. Rst strain with accession number NITE P-02628. [5] The composition according to any one of claims [1] to [4], wherein the cell wall-containing substance of methanol-utilizing bacteria comprises at least one of dead cells and bacterial cell walls. [6] A method for increasing the yield of leafy vegetables, comprising the step of inoculating leafy vegetables with a cell wall-containing substance of methanol-utilizing bacteria. [7] A method for cultivating leafy vegetables, comprising the step of inoculating leafy vegetables with a cell wall-containing substance of methanol-utilizing bacteria. [8] The method according to [6] or [7], wherein the inoculation is performed during the 4-8 leaf stage of the leafy vegetables or 1 to 3 weeks before harvest. [9] A method for increasing the yield of fruit vegetables, comprising the step of inoculating fruit vegetables with a cell wall-containing substance of methanol-utilizing bacteria.

[10] A method for cultivating fruit vegetables, comprising the step of inoculating fruit vegetables with a cell wall-containing substance of methanol-utilizing bacteria.

[11] The method according to [9] or

[10] , wherein the inoculation is performed 5 to 10 days before harvest of the fruit vegetables.

[12] A method for increasing the yield of legumes, comprising the step of inoculating legumes with a cell wall-containing substance of methanol-utilizing bacteria.

[13] A method for cultivating legumes, comprising the step of inoculating legumes with a cell wall-containing substance of methanol-utilizing bacteria.

[14] The method according to

[12] or

[13] , wherein the inoculation is performed between 1 and 50 days after heading of the legume plant.

[15] The method according to any one of [6] to

[14] , wherein the methanol-assimilating bacterium is Methylobacterium sp. Rst strain with accession number NITE P-02628.

[16] The method according to any one of [6] to

[15] , wherein the cell wall content of the methanol-assimilating bacteria comprises at least one of dead cells and bacterial cell walls.

[0009] According to the present invention, it is possible to provide a composition for increasing the yield of leafy vegetables, fruit vegetables, and legumes that has a lower environmental impact than conventional methods, a method for increasing the yield of leafy vegetables, fruit vegetables, and legumes, and a method for cultivating leafy vegetables, fruit vegetables, and legumes.

[0010] The following describes in detail embodiments for carrying out the present invention (hereinafter also referred to as "this embodiment"). It should be noted that the present invention is not limited to this embodiment and can be implemented in various modifications within the scope of its gist.

[0011] 1. Yield-increasing composition This embodiment provides a yield-increasing composition for leafy vegetables, fruit vegetables, or legumes, which contains a cell wall-containing substance of methanol-utilizing bacteria. The yield-increasing composition of this embodiment may contain a cell wall-containing substance of methanol-utilizing bacteria as an active ingredient.

[0012] 1-1. Cell wall-containing material of methanol-assimilating bacteria It is preferable that the cell wall-containing material of methanol-assimilating bacteria contains at least one of the dead cells and cell walls of methanol-assimilating bacteria, as this facilitates formulation and quality control and reduces environmental safety risks when used. In this case, the cell wall-containing material of methanol-assimilating bacteria in this embodiment may contain both the dead cells and cell walls of methanol-assimilating bacteria, or it may contain only one of the dead cells or cell walls of methanol-assimilating bacteria. The cell walls of methanol-assimilating bacteria may be derived from either live or dead cells of methanol-assimilating bacteria. The cell wall-containing material of methanol-assimilating bacteria may contain live cells of methanol-assimilating bacteria.

[0013] When the cell wall content of methanol-assimilating bacteria includes dead cells and / or cell walls, the content of dead cells and / or cell walls in the cell wall content of methanol-assimilating bacteria is not particularly limited and can be appropriately selected by those skilled in the art depending on the purpose. The cell wall content of methanol-assimilating bacteria may consist only of dead cells and / or cell walls, or a solvent may be appropriately added to the dead cells and / or cell walls to form the cell wall content.

[0014] The content of the cell wall components of methanol-assimilating bacteria in the yield-increasing composition is not particularly limited and may be any as long as it exerts a yield-increasing effect, and can be appropriately selected by those skilled in the art depending on the purpose.

[0015] 1-2. Methanol-utilizing bacteria Methanol-utilizing bacteria are bacteria belonging to the genus Methylobacterium. They grow on the surface of plants such as perilla and rice, and are known to utilize methanol derived from plants while also supplying vitamins and plant hormones to the plants, thus forming a symbiotic relationship.

[0016] The method for obtaining methanol-assimilating bacteria is not particularly limited, but for example, those isolated from the surface of plant tissue can be used.

[0017] The method for isolating methanol-utilizing bacteria is not particularly limited, but examples include accumulation culture using methanol alone as the carbon source for the culture medium. Specifically, methods include inoculating a washing solution obtained by washing methanol-utilizing bacteria off the surface of a plant into a culture medium, identifying the colonies that have grown, and isolating the methanol-utilizing bacterial colonies, or inoculating a culture solution obtained by culturing a plant in a liquid culture medium into a culture medium, identifying the colonies that have grown, and isolating the methanol-utilizing bacterial colonies.

[0018] The plant tissue used for isolating methanol-utilizing bacteria is not particularly limited as long as it is a part where methanol-utilizing bacteria grow, but can be appropriately selected depending on the purpose. Examples include the stems and hulls of grains. One or more of these tissues may be used.

[0019] The composition of the culture medium and the cultivation conditions for methanol-utilizing bacteria are not particularly limited and can be appropriately selected according to the purpose.

[0020] While methanol-assimilating bacteria are not particularly limited, Methylobacterium sp. Rst strain is preferred. The Rst strain was deposited with the Patent Microorganism Depositary Center of the National Institute of Technology and Evaluation (NITE) (2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan) and was accepted as NITE P-02628 on February 7, 2018.

[0021] 1-3. Cell Walls of Bacterial Cells There are no particular limitations on the method for preparing the cell walls of methanol-assimilating bacteria, but known methods can be appropriately selected. For example, a method can be used in which cultured and recovered bacterial cells are suspended in an aqueous solution containing 50 mM EDTA and 2% SDS, incubated at approximately 37°C for approximately 60 minutes, then centrifuged, 3 M sodium acetate and ethanol are added to the supernatant and mixed, and centrifuged again to prepare pellet-shaped bacterial cell walls. There are no particular limitations on the method for confirming the presence or absence of bacterial cell wall components, and can be appropriately selected depending on the purpose. For example, a method can be used in which polysaccharides are used as an indicator for confirmation using the phenol-sulfuric acid method.

[0022] 1-4. Dead Bacteria The method for preparing dead bacteria of methanol-assimilating bacteria is not particularly limited, but known methods can be appropriately selected depending on the purpose. For example, a method of culturing and sterilizing the cultured bacteria collected in an autoclave can be used. The dead bacteria may also be crushed bacteria. The method of crushing is not particularly limited, but known methods can be appropriately selected depending on the purpose. The degree of crushing is also not particularly limited, but can be appropriately selected depending on the purpose.

[0023] 1-5. The method for preparing live bacterial cells of methanol-assimilating bacteria is not particularly limited, and known methods can be appropriately selected according to the purpose, for example, a method of recovering cultured bacterial cells.

[0024] 1-6. Other components for increasing yield may further contain the cell wall contents of methanol-assimilating bacteria and other components. The other components are not particularly limited and can be appropriately selected depending on the purpose, and may further contain, for example, components that have a yield-increasing effect.

[0025] The yield-increasing composition may contain a cell wall substance of methanol-assimilating bacteria in combination with one or more agriculturally acceptable components. Agriculturally acceptable components are not particularly limited, but examples include solvents, carriers, excipients, binders, solubilizers, thickeners, emulsifiers, dispersants, suspending agents, spreading agents, penetrating agents, stabilizers, swelling agents, lubricants, surfactants, oily liquids, buffers, bactericides, antifreezes, defoaming agents, colorants, antioxidants, preservatives, and further active ingredients. Agriculturally acceptable carriers include agriculturally acceptable liquid carriers such as water, mineral oil fractions like kerosene or diesel oil, oils of plant or animal origin, 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), or amines (e.g., N-methylpyrrolidone), or mixtures thereof.

[0026] The content of other components in the cell wall contents of methanol-utilizing bacteria in the yield-increasing composition is not particularly limited and can be appropriately selected depending on the purpose.

[0027] The yield-increasing composition can be used in combination with conventionally known agricultural materials such as pesticides and fertilizers. Examples of agricultural materials include pesticides, fertilizers, soil conditioners, and growing media (potting soil). Specifically, pesticides may include fungicides, insecticides, insecticide-fungicides, acaricides, nematicides, rodenticides, insecticides / repellents, attractants, spreading agents, hormones (growth regulators), herbicides, etc. Fertilizers may include organic fertilizers, chemical fertilizers, liquid fertilizers, solid fertilizers, compound fertilizers, compost, growth promoters, vitality enhancers, calcium supplements, etc. Soil conditioners may aim to improve soil physically, chemically, or biologically, and may also include oxygen supply agents, etc. Furthermore, pesticides, fertilizers, soil conditioners, and growing media may contain components derived from plants, animals, lime, minerals, or microorganisms. It is preferable that the cell wall components of methanol-assimilating bacteria include at least one of the dead cells and cell walls of methanol-assimilating bacteria, in order to allow the yield-increasing composition to be used in combination with a fungicide.

[0028] The form of the yield-increasing composition is not particularly limited and can be appropriately selected depending on the purpose. For example, it may be a solid (e.g., powder or granules) or a liquid (e.g., solution or suspension), and it is preferable to use it in the form of a liquid such as a solution or suspension. When used as a solution, it may be in liquid form, or it may be used as a liquid prepared at the time of application. The dosage form of the yield-increasing composition is also not particularly limited and can be appropriately selected depending on the purpose. For example, it can be formulated in dosage forms such as emulsions, wettable powders, liquids, oils, water-soluble powders, granules, powders, aerosols, pastes, or granules.

[0029] Yield-increasing compositions can be used to increase plant yields.

[0030] 2. Method for increasing yield and cultivation This embodiment provides a method for increasing the yield of leafy vegetables, fruit vegetables, or legumes, and a method for cultivating leafy vegetables, fruit vegetables, or legumes.

[0031] 2-1. Step of inoculating a plant with the cell wall contents of methanol-utilizing bacteria The method of this embodiment includes a step of inoculating a plant with the cell wall contents of methanol-utilizing bacteria.

[0032] There are no specific restrictions on the timing of vaccination; it can be chosen as appropriate depending on the purpose.

[0033] When increasing yield or cultivating leafy vegetables, inoculation is preferably carried out, for example, during the 4-8 leaf stage of the leafy vegetable plant or 1-3 weeks before harvest. Inoculation may also be carried out during the 4-8 leaf stage of the leafy vegetable plant and 1-3 weeks before harvest. When increasing yield or cultivating fruit vegetables, inoculation is preferably carried out during a period when the plant is under stress due to fruit set and nutrients being taken by the fruit. For example, inoculation is preferably carried out 5-10 days before harvest of fruit vegetables. If the plant is a crop that is harvested continuously, in the above, "before harvest" may mean before the start of harvest. When increasing yield or cultivating legumes, inoculation is preferably carried out, for example, between 1 and 50 days after heading of the legume plant, more preferably between 1 and 30 days after heading, and even more preferably between 5 and 20 days after heading. Inoculating at the above preferred times is advantageous in that it can further increase the yield of leafy vegetables, fruit vegetables, and legumes.

[0034] The number of vaccinations may be one or multiple. Similarly, if there are multiple vaccination periods, the number of vaccinations at each period may also be one or multiple.

[0035] The amount of methanol-utilizing bacterial cell wall-containing material inoculated into the cereal plants is not particularly limited and can be appropriately selected depending on the purpose. However, the amount of methanol-utilizing bacterial cell wall-containing material inoculated per plant strain, prepared so that the OD is equivalent to 0.5 in OD600 terms, is preferably 1 mL or more, more preferably 2 mL or more, and even more preferably 3 mL or more. The above preferred inoculation amounts are advantageous in that they can further increase the yield of the cereals. The inoculation amounts may be equal or different at each inoculation.

[0036] The method of inoculation is not particularly limited, as long as the cell wall contents of methanol-utilizing bacteria can be brought into contact with at least a portion of the plant, and can be appropriately selected depending on the purpose. Examples include spraying, coating, and immersion. These may be performed individually or in combination of two or more methods. In particular, it is preferable to spray the plant with a liquid containing the cell wall contents of methanol-utilizing bacteria. Inoculation may be done manually or using machinery. When using machinery, there are no particular limitations, but for example, application (spraying) may be done using a drone.

[0037] The part of the plant to which the cell wall-containing substance of methanol-utilizing bacteria is inoculated is not particularly limited and can be appropriately selected depending on the timing of inoculation. For example, it can be inoculated on seeds, seedlings, or the whole or a part of a mature plant, but inoculation on the leaf surface is preferable. According to the method of this embodiment, even if the inoculation of the cell wall-containing substance of methanol-utilizing bacteria does not reach the rhizosphere, the yield-increasing effect can be obtained by inoculating the above-ground part of the plant. Therefore, it is advantageous because it is simpler than adding it as fertilizer and can be used in combination with pesticides.

[0038] 2-2. Other Processes The other processes are not particularly limited as long as they do not impair the effects of the present invention, and ordinary processes in the cultivation of grains can be appropriately selected according to the purpose. The cultivation of plants may be hydroponics or soil cultivation.

[0039] 2-3. Leafy vegetables, fruit vegetables, legumes, and grains. Leafy vegetables are not particularly limited, but examples include spinach, leeks, cabbage, celery, lettuce, Chinese cabbage, broccoli, komatsuna, bok choy, garland chrysanthemum, and mizuna. Among these, the method of this embodiment is preferably used for spinach and leeks.

[0040] 2-4. Fruiting vegetables The fruiting vegetables are not particularly limited, but examples include tomatoes, cucumbers, eggplants, bell peppers, pumpkins, sweet corn, okra, etc. Among these, the method of this embodiment is preferably used for tomatoes.

[0041] 2-5. Leguminous Plants The leguminous plants are not particularly limited, and examples include soybean, adzuki bean, mung bean, cowpea, kidney bean, lima bean, peanut, pea, and broad bean.

[0042] Hereinafter, the present invention will be described with reference to Production Examples and Test Examples, but the present invention is not limited to these Production Examples and Test Examples in any way.

[0043] Production Example 1: Isolation of methanol-assimilating bacteria Using rice stems as a sample, enrichment culture was carried out in a medium prepared by adding 0.01% yeast extract and 0.5% methanol to NMS medium (Whittenbury, R., K. C. Phillips, and J. F. Wilkinson. 1970. Enrichment, isolation and some properties of methane-utilizing bacteria. J. Gen. Microbiol. 61:205-218.). After the enrichment culture, the obtained culture broth was streaked onto a plate medium with the same composition, and the resulting colonies were isolated to obtain Methylobacterium sp. strain Rst.

[0044] The above Methylobacterium sp. strain Rst has been deposited for deposit with the Patent Microorganisms Depositary Center, National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan), and has been accepted as NITE P-02628 on February 7, 2018.

[0045] Production Example 2: Preparation of cell wall content of methanol-assimilating bacteria <Pre-culture> The Methylobacterium sp. Rst strain obtained in the above Production Example 1 was cultured in 5 mL of Hypho minimal medium (Delaney N. F., Kaczmarek M. E., Ward L. M., Swanson P. K., Lee M. C. & Marx C. J. Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens. PLoS One 8, e62957 (2013)) supplemented with 0.2% sodium succinate, and cultured in a test tube at 28°C for 48 hours.

[0046] <Main Culture> To 100 mL of Hypho minimal medium supplemented with 0.5% methanol, the above pre-cultured culture solution was added so that OD₆₀₀ reached 0.002, and cultured in two 500 mL baffled Erlenmeyer flasks at 28°C and 180 rpm for 72 hours. The OD₆₀₀ at the time of bacterial collection was 2.9 to 3.3.

[0047] <Preparation of cell wall content> After completing the above main culture, cultured bacterial cells were collected by centrifugation. Thereafter, the cells were washed twice with sterile water. The washed bacterial cells were sterilized in an autoclave at 121°C for 20 minutes. The sterilized pellet was diluted with a spreading agent having the following composition so that the OD was equivalent to 0.5 in terms of OD₆₀₀ conversion when prepared into a bacterial solution, thereby obtaining a cell wall content solution. Although a spreading agent is used herein, the spreading agent may or may not be used. -Spreading agent- ·Polyoxyethylene (9) nonyl phenyl ether: 0.2 g / L ·Sodium carboxymethyl cellulose: 1 g / L ※ Prepared with ultrapure water

[0048] Using spinach and Welsh onion as examples of leafy vegetables, tests were conducted to increase the yield of leafy vegetables in the following manner.

[0049] Test Example 1: Spinach <Sampling> In a greenhouse in Fukaya City, Saitama Prefecture, the sample was sprayed on 54 spinach plants in each test plot (see below). The first spraying was done on November 29th, and the second spraying was done on December 8th. The spraying amount was approximately 500 mL per 0.2 a, and it was sprayed onto the leaves using a sprayer. The first and second sprayings were done at the 4-leaf and 8-leaf stages of the spinach, respectively, and the spinach was harvested on December 16th. -Test Plots- ・Control plot: Sprayed with spreading agent only ・Test plot: Sprayed with the cell wall-containing solution prepared in Manufacturing Example 2

[0050] <Analysis of Each Test Plot> The total weight of spinach (54 plants) harvested on December 16th was calculated. The result was 32.6 kg in the control plot, compared to 38.7 kg in the test plot. Furthermore, the plant height and leaf color (SPAD value) of the spinach harvested on December 16th were analyzed. The result was that the average plant height was 29.4 cm in the control plot, compared to 30.5 cm in the test plot. The average SPAD value was 59.2 in the control plot, compared to 61.2 in the test plot. This suggests that the cell wall contents of methanol-utilizing bacteria may be involved in the increased yield of leafy vegetables.

[0051] Test Example 2: Green Onions <Sampling> In a greenhouse in Fukaya City, Saitama Prefecture, the sample was sprayed on 265 green onion plants in each test plot (see below). The first spraying was done on November 11th, the second on November 30th, and the third on December 10th. The spraying rate was approximately 1000 mL per 0.2 a, and it was sprayed onto the leaves using a sprayer. The first and second sprayings were done when the green onions were at the 4-leaf and 8-leaf stages, respectively, and the green onions were harvested on December 17th. -Test Plots- ・Control plot: Sprayed with spreading agent only ・Test plot: Sprayed with the cell wall-containing solution prepared in Manufacturing Example 2

[0052] <Analysis of each test plot> The total weight of the scallions (265 plants) harvested on December 17th was calculated. The result was 6.6 kg in the control plot, compared to 8.2 kg in the test plot. Furthermore, the height of the scallions after the third application was analyzed. The result was that the average height was 44.7 cm in the control plot, compared to 52.0 cm in the test plot. This suggests that the cell wall contents of methanol-utilizing bacteria may be involved in the increased yield of leafy vegetables.

[0053] Test Example 3: As an example of a legume, soybeans were used to test the increase in yield of fruit and vegetable crops as follows.

[0054] <Sample Application> The sample was applied to 1200 soybean plants in each test plot (see below) at a field in Kutchan-cho, Abuta-gun, Hokkaido. One application was made approximately 10 days after heading. The amount applied per plant was approximately 8 mL, and it was sprayed onto the leaves using a sprayer. -Test Plots- ・Control plot: Sprayed with only a spreading agent ・Test plot: Sprayed with the cell wall-containing solution prepared in Production Example 2

[0055] <Analysis of each test plot> The dry grain weight per plant was calculated for the harvested soybeans (1200 plants). As a result, it was 17.6 kg in the control plot, compared to 19.8 kg in the test plot. This suggests that the cell wall contents of methanol-utilizing bacteria may be involved in the increased yield of legumes.

[0056] Test Example 4: Using tomatoes as an example of fruit and vegetable crops, we tested increasing the yield of fruit and vegetable crops as follows.

[0057] <Sampling> Samples were sprayed on approximately 80 tomato plants in each test plot (see below) in a greenhouse in Mukawa-cho, Yufutsu-gun, Hokkaido. The first spraying was done in early July, one week after the start of harvesting. The second spraying was done two weeks after the first spraying, and the third spraying was done two weeks after the second spraying. The amount sprayed per plant was approximately 8 mL, and it was sprayed onto the leaves using a sprayer. Tomato harvesting took place from mid-July to late September. -Test plots- ・Control plot: Sprayed with only a spreading agent ・Test plot: Sprayed with the cell wall-containing solution prepared in Manufacturing Example 2

[0058] <Analysis of each test plot> The total weight of tomatoes (80 plants) harvested for seven consecutive days in late September was calculated. As a result, the control plot yielded 16.52 kg, while the test plot yielded 23.8 kg. This suggests that the cell wall contents of methanol-utilizing bacteria may be involved in the increased yield of fruit and vegetable crops.

[0059] NITE P-02628

[0060] [Supplement based on Rule 26, 02.04.2026]

Claims

1. A composition for increasing the yield of leafy vegetables, containing cell wall material of methanol-utilizing bacteria.

2. A composition for increasing the yield of fruit and vegetable plants, comprising cell wall components of methanol-utilizing bacteria.

3. A composition for increasing the yield of legume plants, comprising cell wall components of methanol-assimilating bacteria.

4. The yield-increasing composition according to any one of claims 1 to 3, wherein the methanol-assimilating bacterium is Methylobacterium sp. Rst strain with accession number NITE P-02628.

5. The composition according to any one of claims 1 to 3, wherein the cell wall-containing material of the methanol-assimilating bacteria comprises at least one of dead cells and bacterial cell walls.

6. A method for increasing the yield of leafy vegetables, comprising the step of inoculating leafy vegetables with the cell wall contents of methanol-utilizing bacteria.

7. A method for cultivating leafy vegetables, comprising the step of inoculating leafy vegetables with a cell wall-containing substance of methanol-utilizing bacteria.

8. The method according to claim 6 or 7, wherein the inoculation is performed during the 4-8 leaf stage of the leafy vegetable plant or during the 1-3 week period before harvest.

9. A method for increasing the yield of fruit and vegetable plants, comprising the step of inoculating fruit and vegetable plants with the cell wall contents of methanol-utilizing bacteria.

10. A method for cultivating fruit and vegetable plants, comprising the step of inoculating fruit and vegetable plants with the cell wall contents of methanol-utilizing bacteria.

11. The method according to claim 9 or 10, wherein the inoculation is performed between 5 and 10 days before harvesting of the fruit and vegetable plants.

12. A method for increasing the yield of legume plants, comprising the step of inoculating legume plants with the cell wall contents of methanol-utilizing bacteria.

13. A method for cultivating legumes, comprising the step of inoculating legumes with a cell wall substance of methanol-utilizing bacteria.

14. The method according to claim 12 or 13, wherein the inoculation is performed between 1 and 50 days after heading of the legume plant.

15. The method according to claim 6, 7, 9, 10, 12, or 13, wherein the methanol-assimilating bacterium is Methylobacterium sp. Rst strain with accession number NITE P-02628.

16. The method according to claim 6, 7, 9, 10, 12, or 13, wherein the cell wall-containing material of the methanol-assimilating bacteria comprises at least one of dead cells and bacterial cell walls.

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