JP6572153B2 - Cultivator and method for evaluating substances that affect plant growth using the same - Google Patents

Description

  The present invention relates to a cultivator for cultivating a plant, a cultivating apparatus, a method for cultivating a plant using the same, and a substance or an organism that affects plant growth, that is, a growth promoting substance, a growth inhibiting substance, an infection protective substance, an infection therapeutic substance, an agrochemical. The present invention relates to a method for evaluating the infectivity and mode of infection of bacteria and viruses.

Hydroponics is known as one method for cultivating plants without using soil. In hydroponics, a plant is cultivated in a state where a plant support such as a sponge, mat, or sheet is immersed in or floated on a plant culture solution (for example, Patent Document 1). Hydroponic cultivation is divided into submerged hydroponics, in which plant roots are grown by immersing them directly in the culture solution, and thin-film hydroponics or capillary hydroponics (floating root hydroponic) in which plant roots are grown in moisture. It is done.
On the other hand, a method is also known in which a culture solution is made into a gel, and plant seeds and aerial parts are grown on the surface of the gel medium while supporting the roots inside.

  Cultivation methods that grow plant roots in moisture can supply the roots with sufficient oxygen for root growth, compared to cultivation methods in which the roots are immersed in the culture solution, and also absorb excess plant culture media. There is an advantage that can be suppressed. In addition, seeds and seedlings can be prevented from sinking into the culture solution and becoming poorly grown, and it is easy to cope with the seedling stage from seeding to germination through germination.

JP 2003-143984 A

Generally, in capillary hydroponics, a culture solution sucked up by capillary action spreads over the whole plant support on the culture solution and is supplied to the plant.
However, non-uniformity in the concentration and composition of the culture solution is likely to occur between the position where the culture solution is sucked up and other positions, and the plant growth tends to vary within a single cultivation device or from one cultivation device to another. There's a problem. Variations in plant growth not only have a significant impact on the stable production of plants, but also when performing multi-condition, compound conditions, and multiple specimen cultivation tests at the same time, It is thought that inconvenience also arises in appropriately evaluating candidate compounds useful for growth promotion and the like.
In the cultivation method using a gel medium, although variation in the growth of such a plant is small, since it is necessary to raise the temperature when the culture solution is made into a gel, it is cumbersome and evaluation of candidate compounds that are vulnerable to heat is Have difficulty. In addition, during the cultivation period, addition of culture fluid components, culture fluid exchange, adjustment of the culture fluid volume, water replenishment, inoculation of bacteria and viruses is impossible, and if this is done forcibly, the plant body, especially the roots will be damaged. Cause non-uniformity.

  Accordingly, the present invention has been made in view of the above-mentioned conventional problems and conditions, and the growth of plants is good, the variation in growth is small, and the plant culture solution and its composition are used during the cultivation period. In addition, the present invention relates to providing a new plant cultivator, a cultivation apparatus, a plant cultivation method using the same, and a method for evaluating substances and organisms that affect plant growth, which can be adjusted and exchanged strictly and easily.

In order to solve the above problem, the present invention according to claim 1
A container body for storing a plant culture solution;
A plurality of oscillators disposed on a flat bottom surface in the container body;
A planter comprising a plant support disposed immediately above the plurality of oscillators,
The plurality of oscillators are at least one selected from the group consisting of a sphere, a cylinder, and an elliptic sphere.
The present invention according to claim 2 is characterized in that the plant support is in the form of a sheet.
The present invention according to claim 3 is characterized in that the plurality of oscillators have the same diameter or short axis length.
Further, the present invention according to claim 4 is characterized in that the number of oscillating bodies arranged on the flat bottom surface of the container body is three or more.
Moreover, the present invention according to claim 5 is characterized in that the cultivator is used in combination with a rocking device.
The present invention according to claim 6
A plant cultivation device comprising a cultivator and a rocking device,
The cultivator includes a container body that contains a plant culture solution, and a plurality of oscillating bodies selected from the group consisting of a sphere, a cylinder, and an elliptic sphere disposed on a flat bottom surface in the container body; A plant support disposed immediately above the plurality of oscillators,
The container body contains a plant culture solution so that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other,
The plurality of oscillating bodies are disposed on a flat bottom surface in the container main body so that the center or the central axis thereof is located on the same plane and can be oscillated. The surface of the plurality of oscillating bodies is transmitted along with the oscillation of the plurality of oscillating bodies by the oscillating device, and the plurality of oscillating bodies and the plant support are in contact with each other through the plurality of contacts or tangents. It is characterized by being supplied to a plant support.
The present invention according to claim 7 provides
A plant cultivation method using a cultivator and a rocking device,
The cultivator comprises a container main body, at least one type of a plurality of rocking bodies selected from the group consisting of a sphere, a cylindrical body, and an elliptic sphere, and a plant support.
Disposing the plurality of oscillating bodies on the flat bottom surface in the container main body so that the center or the central axis thereof is located on the same plane and swingable;
Disposing the plant support directly on the plurality of oscillators;
Storing the plant culture solution in the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other;
Placing a plant on the plant support;
The plant culture solution is transmitted to the surface of the rocking body along with the rocking of the rocking bodies by the rocking device and supplied to the plant support through the plurality of contacts or tangents. And the step of cultivating the plant.
The present invention according to claim 8 provides:
A method for evaluating substances and / or organisms that affect plant growth using a cultivator and a rocking device,
The cultivator comprises a container main body, at least one type of a plurality of rocking bodies selected from the group consisting of a sphere, a cylindrical body, and an elliptic sphere, and a plant support.
Disposing the plurality of oscillating bodies on the flat bottom surface in the container main body so that the center or the central axis thereof is located on the same plane and swingable;
Disposing the plant support directly on the plurality of oscillators;
A plant culture solution containing substances and / or organisms that affect plant growth is placed on the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other. A step of accommodating in,
Placing a plant on the plant support;
The plant culture solution containing the substance that affects plant growth and / or organisms is transmitted along the surface of the rocking body as the rocking body is swung by the rocking device, and the plurality of contacts or Supplying the plant support through a tangent and cultivating the plant;
And a step of evaluating the growth state of the cultivated plant.
The present invention according to claim 9
A method for evaluating substances and / or organisms that affect plant growth using a cultivator and a rocking device,
The cultivator comprises a container main body, at least one type of a plurality of rocking bodies selected from the group consisting of a sphere, a cylindrical body, and an elliptic sphere, and a plant support.
Disposing the plurality of oscillating bodies on the flat bottom surface in the container main body so that the center or the central axis thereof is located on the same plane and swingable;
Disposing the plant support directly on the plurality of oscillators;
Placing a plant pretreated with an organism that affects plant growth on said plant support;
Storing the plant culture solution containing a substance that affects plant growth in the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other. When,
A plant culture solution containing a substance that affects plant growth is transmitted along the surface of the rocking body as the rocking body is swung by the rocking device, and the plant is transmitted through the plurality of contacts or tangents. Feeding the support and cultivating the plant;
And a step of evaluating the growth state of the cultivated plant.

  According to the present invention, the plant grows well, the variation in the effects of substances and organisms that affect plant growth, the variation in growth is small, and the plant culture solution and its composition are adjusted according to the growth status of the plant, A replaceable plant grower is provided. In addition, with the use of the cultivator of the present invention, it is possible to carry out multi-condition, multi-condition, multi-specimen cultivation tests at once in a short period of time with labor-saving, space-saving and low running cost, It is possible to simply and appropriately evaluate the effects of organisms, optimal concentrations, and the like.

It is a figure explaining the basic composition of the cultivation machine concerning one embodiment of the present invention. It is a figure explaining the cultivation method of the plant using a cultivator. (A) It is a figure which shows the state which has arrange | positioned the cylindrical body in the container main body. (B) It is a figure which shows the state which has arrange | positioned the spherical body in the container main body. It is a figure which shows the plant individual weight obtained by each cultivation method. It is a figure which shows the change of the chlorophyll amount of Arabidopsis thaliana by ALA administration. It is a figure which shows the growth state of Arabidopsis thaliana on the 19th day of cultivation. It is a figure which shows the growth state of Arabidopsis thaliana on the 19th day of cultivation. It is a figure which shows the growth state of the watercress of the cultivation 12th day. It is a figure which shows the growth state of the lettuce of the cultivation 12th day. It is a figure which shows the growth state of the basil of the cultivation 16th day. It is a figure which shows the growth state of the parsley of the cultivation 28th day.

  Embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is a diagram illustrating a basic configuration of a cultivator according to an embodiment of the present invention. In FIG. 1, U is a cultivator, P is a container main body for storing a plant culture solution, S is a rocking body, and B is a plant support.
As shown in FIG. 1, the cultivator U has a plurality of oscillating bodies S disposed on the flat bottom surface inside the container body P, and a plant support B is disposed immediately above the plurality of oscillating bodies S. Yes.

The container body P for storing the plant culture solution is not particularly limited, and examples thereof include a cell tray, a seedling pot, a water tank, a petri dish, and a plate having an opening at the upper end. The inner bottom surface of the container body P is planar.
The container main body P is preferably a round petri dish, a plate-type petri dish, or a plate from the viewpoint that it can be made compact when multiple conditions, complex conditions, and multiple sample plant cultivation tests are performed at once. In particular, in the present invention, it is more preferable to use a multi-hole plate (flat bottom) having a plurality of holes (wells, bottomed holes), for example, 6 holes or 12 holes, for accommodating a plant culture solution. The well diameter and well depth are not particularly limited.
As a material of the container main body P, for example, a plastic, such as glass, stainless steel, polystyrene, polypropylene, or polycarbonate is preferable.

The shape of the plurality of oscillating bodies S is a sphere, a cylinder, or an elliptic sphere. The shape of the rocking body S may be any shape as long as it has a substantially spherical appearance as a whole even if the surface has irregularities and undulations like a golf ball. The ellipsoidal sphere may be either a long sphere with the major axis of the ellipse as the rotation axis or a flat sphere with the minor axis as the rotation axis, but is preferably an oblong.
The shape of the oscillating body S is preferably a sphere or a cylindrical body from the viewpoint of increasing the uniformity of the culture solution and reducing the variation in the effects of substances and organisms that affect plant growth and the variation in plant growth. A sphere is preferable.

  As long as a plurality of the oscillating bodies S can be accommodated in the container body P, those having a suitable size can be used. For example, the diameter of the sphere or cylinder, or the length of the minor axis of the elliptic sphere is 5 to 10 mm, preferably 7 to 9 mm. The length of the cylinder or the major axis of the elliptic sphere is 8 -15 cm, preferably 10-12 cm.

  The material of the oscillating body S is not particularly limited as long as it can attach a plant culture solution by surface tension, and examples thereof include plastic balls such as polystyrene and polypropylene, glass balls, stainless steel balls, steel balls, rubber balls, and the like. Can be mentioned. From the viewpoint of handleability, a plastic sphere is preferable.

  The number of the oscillating bodies S is plural, that is, two or more. The number of the rocking bodies S is preferably 3 or more from the viewpoint of enhancing the uniformity of the culture solution and reducing the variation in the effects of substances and organisms that affect plant growth and the variation in plant growth. The upper limit is not particularly limited, and is preferably 100 or less, more preferably 80 or less, and even more preferably 50 or less, although it varies depending on the flat bottom area in the container body P and the size of the rocking body S.

The plurality of oscillating bodies S have the same diameter or short axis length from the viewpoint of increasing the uniformity of the culture solution and reducing the variation in the effects of substances and organisms that affect plant growth and the variation in plant growth. It is preferable that
Moreover, although the same shape or different shapes may be sufficient as the combination of the rocking bodies S, it is preferable to combine the same shape from the same viewpoint.

The plant support B is a support that can support a plant. As will be described later, the plant absorbs the culture solution through the plant support B and grows. Therefore, the plant support B has a property of absorbing or permeating a plant culture solution.
The plant support B preferably does not adsorb the components of the plant culture solution. Furthermore, the plant support B preferably has a small pore size and has a root-preventing property that blocks plant root entry from the viewpoint of high-throughput screening of substances and organisms that affect plant growth. When plant roots do not enter the plant support and grow on the plant support with sufficient culture medium, not only will the growth be good, but the support and plant will be easy to separate after growth without damaging the roots. Plant transplantation, plant weight and length measurement, etc. can be performed accurately, easily and safely. In addition, the plant culture solution and its composition adjustment and exchange according to the growth of the plant, that is, addition of the culture solution, exchange of the culture solution, inoculation of bacteria and viruses, and adjustment of the culture solution amount can also be performed on the support It can be carried out easily or safely while it is grown.
From this point of view, the plant support B is preferably in the form of a sheet capable of maintaining flatness, more preferably cloth, paper, non-woven fabric, etc., and further preferably paper, non-woven fabric.

  Examples of the material for the plant support B include hydrophilic fibers such as cellulose fibers, nylon fibers, polyester fibers, polyolefin fibers, polyvinyl alcohol fibers, and polyacrylonitrile fibers.

Two or more plant supports B may be arranged immediately above the plurality of rocking bodies S, but the generation of a nonuniformity due to air mixing between the stacked supports and the prevention of uniform component diffusion due to deviation. From this point of view, it is preferable that one is disposed immediately above the plurality of oscillators S.
The plant support B may be disposed in contact with a plurality of rocking bodies S at a plurality of contacts or tangents. However, the plant support B improves the uniformity of the culture solution and has the effect of substances and organisms that affect plant growth. From the viewpoint of reducing variations and variations in plant growth, it is preferable to arrange them over the entire surface of the container body P.

Furthermore, the cultivator U may include a lid that is detachably attached. By attaching a lid to the upper end opening of the cultivator and keeping the cultivator in a hermetically sealed condition, it can be an aseptic environment or an environment in the presence of only specific bacteria.
The lid of the cultivator U can be appropriately selected according to the cultivator, and examples thereof include a resin cover, a plate seal, Saran wrap (registered trademark), a transparent vinyl sheet, and the like.

Next, a method for cultivating a plant using the cultivator U configured as described above will be described with reference to FIG. 2, and a rocking device (not shown) is used in combination for plant cultivation. That is, a plant cultivating apparatus including a cultivator U and a swinging device that swings the cultivator U is used. As shown in FIG. 2, when the cultivator U is oscillated by the oscillating device, a plurality of oscillating bodies S in the cultivator U oscillate, and the plant culture solution 11 travels along the surface of the oscillating body. Since it is supplied to the plant support B arranged just above the rocking body, the plant 12 can be cultivated on the plant support B. In FIG. 2, reference numeral 10 denotes a contact or tangent where the plurality of rockers S and the plant support B are in contact.
Examples of the swing device include a known shaker.

In plant cultivation, first, a plurality of oscillating bodies S are arranged on the flat bottom surface in the container body P so that their centers or central axes are located on the same plane and can be oscillated. That is, it arrange | positions so that rocking bodies may not mutually overlap on the flat bottom face in the container main body P. FIG. In the elliptical sphere, it is preferable that the long axis is a central axis, and the central axis (long axis) is located on the same plane and is swingable.
A plurality of oscillators S are provided on the flat bottom surface of the container body P from the viewpoint of improving the uniformity of the culture solution and reducing the variation in the effects of substances and organisms that affect plant growth and the variation in plant growth. It is preferable to arrange. FIG. 3A is a diagram in which seven cylinders having the same diameter are arranged in a maximum number so that the central axes are located on the same plane and can be swung, and the eight cylinders can be swung. This indicates a state where it cannot be placed. FIG. 3B is a diagram in which five spheres having the same diameter are arranged in such a manner that the center point is located on the same plane and can be swung. Six spheres cannot be swung. Is shown.

  A plant support B is disposed immediately above the plurality of oscillators S. At this time, the plurality of rocking bodies S and the plant support B are in contact with each other at a plurality of contacts or tangents 10.

Further, the plant 12 is placed on the plant support B. It does not specifically limit as a kind of plant 12, Various flowers, vegetables, a herb etc. are mentioned. The plant 12 may have been subjected to pretreatment such as contacting a substance or organism that affects plant growth.
The plant may be seeded with seeds or seedlings may be transplanted. It is preferred to sow plant seeds on the plant support.

In the container body P, the plant culture solution 11 is stored such that the liquid level is lower than the plurality of contacts or tangents 10 where the plurality of rocking bodies S and the plant support B are in contact. The liquid level of the plant culture solution 11 may be lower than that of the contact or tangent line 10, but a necessary and sufficient amount of the culture solution is supplied to the plant support, but seeds and seedlings on the plant support are cultured. From the viewpoint of preventing sinking in the liquid, when the distance from the flat bottom surface in the container body P to the contact or tangent line 10 where the rocking body S and the plant support B are in contact is 1, the flat bottom surface in the container body P It is preferable that the distance is 0.9 or less, further 0.8 or less, and further 0.5 to 0.7.
The plant culture solution 11 is obtained by adding water, a culture solution generally used for plant cultivation, and various substances and organisms that affect plant growth.

  In this way, the plant culture solution 11 accommodated in the container main body P is transmitted to the surface of the oscillating body with the oscillation of the plurality of oscillating bodies S by oscillating the cultivator U by the oscillating device. The plant support B is supplied through a plurality of contacts or tangents 10, and the plant 12 on the plant support B is cultivated. Since there are multiple contact points or tangents to which the plant culture solution is supplied, the culture solution including substances and organisms that affect plant growth is uniformly and stably supplied to the entire plant support and the plants supported by the plant support. Thus, the plant grows well in a short period of time, and the growth is uniform among plant individuals regardless of the cultivation position, thereby reducing variation. In addition, when a plant is cultivated in each well of the multi-hole plate, the growth of the plant is even between the wells and the variation is reduced.

Oscillation of the oscillating body S by the oscillating device is performed by supplying the plant culture solution 11 attached by surface tension to the plant support B from a plurality of contacts or tangents 10 where the plurality of oscillating bodies S and the plant support B are in contact. In the case of a sphere, it is a turning motion and a rotation motion along the circular arc of the sphere. Further, in the case of a cylinder or an elliptic sphere, a reciprocating motion and a rotation motion along an arc around the central axis of the cylinder or the elliptic sphere. Note that the rotation of the rocking body S may be less than 360 degrees, and it is not always necessary to make one rotation.
The swing condition can be set as appropriate, but in the case of a sphere, it is preferably horizontal swinging at 6 to 180 r / min. In the case of a cylindrical body or an elliptical sphere, the swaying width is preferably 1 mm to 10 cm.

In plant cultivation, temperature, humidity, light conditions, cultivation days, and the like can be appropriately set according to the type of plant. You may grow in the incubator etc. which were set up on fixed conditions.
In the cultivator of the present invention, if the oscillating conditions of a plurality of oscillating bodies are the same, the plant culture solution is uniformly supplied to the plant support and the plants supported by the cultivating apparatus. The growth is uniform not only between the plants but also between the individual plants for each cultivator, with little variation and uniform growth. Therefore, if the cultivation device of the present invention is used, it is possible to cultivate a plant with high quality, accurately evaluate growth, and the like.
Moreover, it is possible to simultaneously perform a multi-condition, multi-condition, multi-sample cultivation test in a short period of time, labor saving, space saving, and low running cost. For example, as will be described later in Examples, it is possible to simply and appropriately evaluate substances useful for plant growth, usefulness of organisms, optimum concentrations, and the like.

Next, a method for evaluating substances and / or organisms that affect plant growth using the cultivator U of the present invention will be described. Also in the evaluation method, as described above, a rocking device is used in combination.
In the method for evaluating a substance and / or organism that affects plant growth, a container culture solution 11 containing a substance and / or organism that affects plant growth is accommodated in the container body P. In the same manner as described above, the culture medium 11 is transmitted along the surface of the oscillating body as the plurality of oscillating bodies S are oscillated by the oscillating device, and the plurality of oscillating bodies S and the plant support B are in contact with each other. The plant 12 is cultivated by supplying the plant support B through the contact or tangent line 10. When plants are cultivated in this way, the variation in the effects of substances and organisms that affect plant growth contained in the culture solution is reduced, and the effects of substances and organisms that affect plant growth in a short period of time, the optimum concentration, etc. can be simplified. In addition, it is possible to evaluate appropriately.
In addition, in the method for evaluating substances and / or organisms that affect plant growth, the plant 12 is pre-treated in advance, for example, by contacting the plant 12 with organisms that affect plant growth. The plant culture solution 11 containing the substance to be stored is contained, and the culture solution 11 is transmitted to the surface of the rocking body along with the rocking of the rocking bodies S by the rocking device in the same manner as described above. The plant 12 may be cultivated by supplying it to the plant support B through a plurality of contacts or tangents 10. Thereby, it is possible to simply and appropriately evaluate the effects of substances and organisms that affect plant growth.

  Examples of substances and organisms that affect plant growth that can be evaluated using the cultivator of the present invention include plant growth, for example, plant weight, increase in the number of leaves / stems, elongation / expansion of plant bodies, flower bud formation, fruit set, etc. Organic / inorganic compounds and organisms that affect the environment. The types include, for example, amino acids, sugars, lipids, nucleic acids, proteins, nitrogen, phosphorus, potassium, heavy metals, plant pathogens, soil bacteria, mycorrhizal fungi, rhizobia, viruses, or combinations thereof. Substances and organisms that affect plant growth include any candidate compounds and organisms in addition to those whose usefulness and toxicity are estimated and expected.

  In the evaluation of the growth state of cultivated plants, for example, the growth state of cultivated plants in the presence of substances or organisms that affect plant growth and the growth state of cultivated plants in the absence of substances or organisms that affect plant growth. In comparison, if the growth of cultivated plants in the presence of substances or organisms that affect plant growth is improved, the substances or organisms can be evaluated and selected as substances or organisms useful for promoting plant growth. . Conversely, when the growth of the cultivated plant is suppressed, the substance or organism can be evaluated and selected as a substance or organism useful for suppressing plant growth or a substance or organism toxic to plants. In addition, by changing the concentration of substances and organisms that affect plant growth in the culture solution and comparing the growth state of cultivated plants at each concentration, the optimum concentration of substances and organisms that affect plant growth can be evaluated. Can do.

  Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.

The following materials and plants were used.
[Culture medium]
MS medium (pH; 5.7-5.8, composition is shown in Table 1)
Hyponex medium (stock solution, Hyponex)
Hyponica medium (liquid fertilizer, manufactured by Kyowa Co., Ltd.)
[container]
12 hole plate (flat bottom, material: polystyrene, plate outer dimension: 85.4 mm x 127.6 mm, well diameter: φ22.1 mm)
[sphere]
Polypropylene sphere (diameter: about 8mm)
[Plant support]
Hydrophilic paper (filter paper, material: Advantech filter paper No. 514A, thickness 0.32 mm, cellulose fiber), cut into a circle with a diameter of 20 mm for use.
[plant]
Arabidopsis thaliana (wild strain, T-DNA insertion mutant, genetically engineered strain, genome editing strain)
Watercress (Watercress)
Lettuce (Salinas 88)
Basil (sweet basil)
Parsley (Italian parsley)

Test example 1
(1) The following cultivation plate was produced.
[Gel plate]
2 mL each of autoclaved sucrose-free MS medium (containing 0.6% gellan gum) was poured into each well of a sterilized 12-well plate at about 50 ° C., cooled and solidified to form a gel plate. In FIG. 4, it was expressed as gel (cont).

[Liquid plate]
2 mL of sterilized sucrose-free MS medium was injected into each well of a sterilized 12-well plate to prepare a liquid plate. In FIG. 4, it was described as no support.

[Funnel plate]
A filter paper was cut out in a circular shape and cut from the outer periphery toward the center to form a funnel shape. 1 mL of sterilized sucrose-free MS medium was injected into each well of a sterilized 12-well plate, and funnel-shaped filter papers were placed one by one to form a funnel plate. In this plate, the medium is supplied to the entire filter paper through the center of the filter paper by capillary action. In FIG. 4, it was described as funnel.

[Bridge plate]
The filter paper was cut into a rectangle and folded at two places to form a bridge. 1.5 mL of sterilized sucrose-free MS medium was injected into each well of a sterilized 12-well plate, and a bridged filter paper was installed one by one to form a bridge plate. In this plate, the medium is supplied to the entire filter paper through the bridge portion of the filter paper folded by capillary action. In FIG. 4, it was written as bridge.

[Sphere plate]
Five sterilized polypropylene spheres were placed in each well of a sterilized 12-well plate so as not to overlap each other, and 1200 μL of sucrose-free MS medium was injected. At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate. In FIG. 4, it was described as sphere.

(2) The seeds of Arabidopsis thaliana are aseptically sown on the culture medium, culture solution or filter paper of each cultivation plate prepared in (1) above, the lid is closed, and the vinyl tape and surgical tape are placed around the medium. In a dark state, the plant was cultivated for 2 weeks under continuous irradiation conditions of 23 μC and 70 μE of daylight white fluorescent light (N = 6 or 7).
During the cultivation period, the sphere plate was shaken horizontally at 60 r / min using a shaker (Taitech Co., Ltd. shaker, the same applies hereinafter).

(3) After cultivation, the weight of each plant was measured. Evaluation is based on individual weight and C.I. V. Comparison was made by (Coefficient of variation, coefficient of variation). C. V. The following formula was used for calculation.
C. V. (%) = (Standard deviation) / (average) × 100
The results are shown in Table 2 and FIG.

As shown in Table 2 and FIG. 4, the median value was 3.50 g as a result of cultivation on a gel plate. V. The value was 28.8%.
In the liquid plate, the median was as low as 0.45 g, and seeds and seedlings were submerged and could not be exposed to the air, so photosynthesis was not possible, and it was confirmed that they could not grow on MS medium without sucrose. In addition, the funnel plate grew slightly larger at a median of 1.15 g, but the degree of growth was about 1/3 that of the gel plate. The bridge plate grew to a median of 3.60 g, which was about the same weight as the gel plate. V. The value was 59.1%, and the variation among individuals was large between each well.
On the other hand, with the spherical plate, the median is 3.70 g, which is equivalent to or higher than the gel plate, and C.I. V. The value is 37.9% and the C.I. V. Compared to the value, it was confirmed that the individual variation between each well could be suppressed.

Test example 2
(1) Six Arabidopsis thaliana grains are placed inside 5 circular petri dishes, and 6 seeds are aseptically seeded on a filter paper sucked with 10 mL of MS medium. After vernalization at 4 ° C. for 24 hours, circular The petri dish was moved to the cultivation room, and aseptic cultivation was started under continuous irradiation conditions of 23 ° C. and 70 μE under daylight white fluorescent lamp (cultivation day 0).

(2) Place 5 polypropylene spheres in each well of a 12-well plate so as not to overlap each other, and add 5-200-aminolevulinic acid (ALA) hydrochloride solution (hydroxylized) so that the final concentration shown in FIG. PH was adjusted from 5.5 to 5.9 with potassium). At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate.
On the 6th day from the start of cultivation, two seedlings were transplanted from the circular petri dish into each well of the sphere plate, the lids were closed, sealed with vinyl tape and surgical tape, and further irradiated continuously at 23 ° C under daylight white fluorescent light at 70 µE. Under aseptic conditions, aseptic cultivation was performed using a shaker for 16 days with horizontal swirling at 60 r / min (N = 4).

  On the 22nd day from the start of cultivation, the plant was removed from the well and the amount of chlorophyll was measured. The amount of chlorophyll was measured by spectroscopy using dimethylformamide extraction. The results are shown in FIG. In addition, 5-aminolevulinic acid hydrochloride is reported as one of the substances which increase chlorophyll in plants and promote growth.

  As shown in FIG. 5, the amount of chlorophyll in the plant increased as the concentration of 5-aminolevulinic acid hydrochloride added increased, and decreased when given too much. It was confirmed that evaluation was possible.

Test example 3
(1) In the same manner as in Test Example 2, 100 seeds of Arabidopsis thaliana were aseptically seeded on a filter paper sucked with the MS medium in 5 round petri dishes, and subjected to vernalization at 4 ° C. for 24 hours. The circular petri dish was moved to the cultivation room, and aseptic cultivation was started under continuous irradiation conditions of 23 [deg.] C. and 70 [mu] E under daylight white fluorescent light (cultivation day 0).

(2) Five polypropylene spheres were placed in each well of a 12-well plate so as not to overlap each other, and two kinds of MS medium, hyponex medium and hyponica medium were appropriately diluted and mixed at various concentrations shown in Table 3. A total of 1200 μL of medium was injected. At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate.
On the 6th day from the start of cultivation, two seedlings were transplanted from the circular petri dish into each well of the sphere plate, the lids were closed, sealed with vinyl tape and surgical tape, and continuous irradiation conditions at 23 ° C and 70 μE under daylight white fluorescent light Then, aseptically cultivated under the same conditions (N = 2) while shaking horizontally at 60 r / min using a shaker for 13 days.
Photographs were taken on the 19th day from the start of cultivation, and the plants were taken out of the wells and the plant weight was measured. Table 3 shows the average weight of the plants in each well, and FIG. 6 shows the growth state of Arabidopsis on the 19th day of cultivation.

Test example 4
(1) In the same manner as in Test Example 2, 100 seeds of Arabidopsis thaliana were aseptically seeded on a filter paper sucked with the MS medium in 5 round petri dishes, and subjected to vernalization at 4 ° C. for 24 hours. The circular petri dish was moved to the cultivation room, and aseptic cultivation was started under continuous irradiation conditions of 23 [deg.] C. and 70 [mu] E under daylight white fluorescent light (cultivation day 0).

(2) A medium in which five polypropylene spheres are placed so as not to overlap each well of a 12-well plate, and a Hyponex medium and a Hyponica medium diluted with various concentrations as shown in Table 4, or an MS medium Was diluted and the concentration was changed to two stages, and further, Hyponex medium and Hyponica medium were diluted, and a medium covered with various concentrations was injected in a total amount of 1200 μL. At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate.
On the 5th day from the start of cultivation, two seedlings were transplanted from the circular petri dish into each well of the sphere plate, the lids were closed, and sealed with vinyl tape and surgical tape. Then, the plant was cultivated under the same conditions while shaking horizontally at 60 r / min using a shaker for 14 days (N = 2).
Photographs were taken on the 19th day from the start of cultivation, and the plants were taken out of the wells and the plant weight was measured. Table 4 shows the average weight of the plants in each well, and FIG. 7 shows the growth state of Arabidopsis on the 19th day of cultivation.

As shown in Table 3 and Table 4, Arabidopsis thaliana grew well by spherical plate cultivation. Moreover, as shown in FIG.6 and FIG.7, the dispersion | variation in growth was small between the plant individual | organism | solids in a well, and it was able to evaluate in about 3 weeks after sowing.
Furthermore, it was confirmed from Test Examples 3 and 4 that the combined effect of two or three agents against Arabidopsis thaliana can be studied very quickly and easily.

Test Examples 5-7
(1) Aseptically seeded on a filter paper sucked with 10 mL of MS medium in one circular petri dish each of 50 watercress, lettuce, and basil, transferred to the cultivation room, and started cultivation (Day 0 of cultivation) ). Cultivation was performed under continuous irradiation conditions of 23 [deg.] C. and 70 [mu] E under a daylight fluorescent lamp.

(2) Place 5 polypropylene spheres in each well of the 12-well plate so that they do not overlap each other, and inject a total of 1200 μL of medium containing MS medium concentration and hyponica medium concentration as shown in Tables 5-7. did. At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate.
On the 3rd day from the start of cultivation, two seedlings were transplanted from the circular petri dish into each well of the sphere plate, the lids were closed, sealed with vinyl tape and surgical tape, and continuous irradiation conditions at 23 ° C and 70μE under daylight white fluorescent light Then, the plant was cultivated with horizontal shaking at 60 r / min using a shaker (N = 2).
On the 12th day from the start of cultivation, watercress and lettuce were photographed, and the plants were removed from the wells and the plant weight was measured. Basil was photographed on the 16th day from the start of cultivation, and the plant weight was taken out from the well. Tables 5 to 7 show the average weight of the plants in each well, and FIGS. 8 to 10 show the growth state of watercress, lettuce or basil on the 12th or 16th day of cultivation.

Test Example 8
(1) 50 seeds of parsley were aseptically seeded on a filter paper in which 10 mL of MS medium in a circular petri dish was sucked, and cultivation was started under continuous irradiation conditions of 23 μC and 70 μE under daylight white fluorescent light (cultivation) Day 0).

(2) Five polypropylene spheres were placed in each well of a 12-well plate so as not to overlap each other, and as shown in Table 8, a total of 1200 μL of medium filled with MS medium concentration and hyponica medium concentration was injected. At this time, the liquid level of the culture solution was at a height of 1/2 or more and 2/3 or less of the diameter of the polypropylene sphere. Next, one piece of filter paper cut out in a circle was placed immediately above the five polypropylene spheres to form a spherical plate.
On the 14th day from the start of cultivation, two seedlings were transplanted from the circular petri dish into each well of the sphere plate, the lid was closed, sealed with vinyl tape and surgical tape, and continuous irradiation conditions at 23 ° C and 70μE under daylight white fluorescent light Then, cultivation was continued using a shaker for 14 days at 60 r / min with horizontal swirling (N = 2).
Photographs were taken on the 28th day from the start of cultivation, and the plants were taken out of the wells and the plant weight was measured. Table 8 shows the average weight of the plants in each well, and FIG. 11 shows the growth state of parsley on the 28th day of cultivation.

As shown in Tables 5 to 8, it was confirmed that the herb grows as well as Arabidopsis thaliana in a short period of time by spherical plate cultivation. Moreover, as shown in FIGS. 8-11, the dispersion | variation in growth was small between the plant individuals in a well.
Furthermore, from Test Examples 5 to 7 and 8, the combined effect of the two agents on each plant can be examined very quickly and easily, and the optimum concentration for each plant was confirmed.

U ... cultivator P ... container body S for storing plant culture medium ... rocking body B ... plant support 10 ... contact or tangent 11 where rocking body and plant support are in contact Plant culture solution 12. Plant

Claims (9)

A cultivator used in combination with a rocking device ,
A container body for storing a plant culture solution;
A plurality of oscillating bodies that are arranged on a flat bottom surface in the container body and oscillate and rotate by oscillating the cultivator with an oscillating device, the shape of which is a sphere, a cylinder, or an elliptic sphere ,
A cultivator comprising a plant support disposed directly above the plurality of oscillators .   The cultivator according to claim 1, wherein the plant support is in a sheet form.   The cultivator according to claim 1 or 2, wherein the plurality of oscillators have the same diameter or short axis length.   The cultivator according to any one of claims 1 to 3, wherein the number of rocking bodies disposed on the flat bottom surface of the container body is three or more. Wherein the plurality of the rocking body flat bottom surface in the container body, cultivation device according to any one of claims 1 to 4 in which the center or center axis Ru is arranged to be positioned on the same plane. A plant cultivation device comprising a cultivator and a rocking device,
The cultivation comprises: a container body for containing the culture medium of the plant, the arranged flat bottom surface of the container body, swings rotated by oscillation of the cultivation device according to the rocking device, shaped spherical, cylindrical, or A plurality of oscillating bodies that are elliptical spheres , and a plant support disposed directly above the plurality of oscillating bodies,
The container body contains a plant culture solution so that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other,
The plurality of oscillating bodies are disposed on a flat bottom surface in the container main body so that the center or the central axis thereof is located on the same plane and are capable of swinging and rotating , and the plant culture solution in the container main body Is transmitted to the surface of the plurality of oscillating bodies by the oscillation of the cultivator by the oscillating device, and the plant supports are connected to the plant support through the plurality of contacts or tangents where the plurality of oscillating bodies contact the plant support. Supply plant cultivation equipment. A plant cultivation method using a cultivator and a rocking device,
The cultivator comprises a container body , a plurality of oscillating bodies whose shape is a sphere, a cylinder, or an ellipsoidal sphere, which is oscillated and rotated by oscillating the cultivator by an oscillating device , and a plant support. ,
Disposing the plurality of oscillating bodies on the flat bottom surface in the container body so that the centers or the central axes thereof are located on the same plane and swingably rotatable ;
Disposing the plant support directly on the plurality of oscillators;
Storing the plant culture solution in the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other;
Placing a plant on the plant support;
Cultivating the plant by supplying the plant culture solution to the surface of the oscillating body by the oscillation of the cultivator by the oscillating device and supplying the plant support through the plurality of contacts or tangents; A method for cultivating a plant. A method for evaluating substances and / or organisms that affect plant growth using a cultivator and a rocking device,
The cultivator comprises a container body , a plurality of oscillating bodies whose shape is a sphere, a cylinder, or an ellipsoidal sphere, which is oscillated and rotated by oscillating the cultivator by an oscillating device , and a plant support. ,
Disposing the plurality of oscillating bodies on the flat bottom surface in the container body so that the centers or the central axes thereof are located on the same plane and swingably rotatable ;
Disposing the plant support directly on the plurality of oscillators;
A plant culture solution containing substances and / or organisms that affect plant growth is placed on the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other. A step of accommodating in,
Placing a plant on the plant support;
A plant culture solution containing substances and / or organisms that affect the plant growth is transmitted along the surface of the rocking body by rocking of the cultivator by the rocking device, and the plant support is supported through the plurality of contacts or tangents. Feeding the body and cultivating the plant;
A method for evaluating substances and / or organisms that affect plant growth, comprising the step of evaluating the growth state of cultivated plants. A method for evaluating substances and / or organisms that affect plant growth using a cultivator and a rocking device,
The cultivator comprises a container body , a plurality of oscillating bodies whose shape is a sphere, a cylinder, or an ellipsoidal sphere, which is oscillated and rotated by oscillating the cultivator by an oscillating device , and a plant support. ,
Disposing the plurality of oscillating bodies on the flat bottom surface in the container body so that the centers or the central axes thereof are located on the same plane and swingably rotatable ;
Disposing the plant support directly on the plurality of oscillators;
Placing a plant pretreated with an organism that affects plant growth on said plant support;
Storing the plant culture solution containing a substance that affects plant growth in the container body such that the liquid level is lower than a plurality of contacts or tangents where the plurality of oscillators and the plant support are in contact with each other. When,
A plant culture solution containing a substance that affects plant growth is supplied to the plant support through the plurality of contacts or tangential lines, along the surface of the rocking body by rocking the cultivator by the rocking device. Cultivating plants,
A method for evaluating substances and / or organisms that affect plant growth, comprising the step of evaluating the growth state of cultivated plants.