JPH0677516B2 - Plant tissue culture equipment - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial field of application] The present invention irradiates strong light and distributes air to promote metabolism of plants, accelerate growth, and increase temperature in a culture device by strong light irradiation. The present invention relates to a plant tissue culture device that prevents

[Conventional technology]

In so-called tissue culture, which is separated from organs and tissues of plants, cells and plants, and aseptically cultivated under appropriate culture conditions to regenerate solids that have complete functions as plants, so-called sterile Conditions must be maintained. At the same time, since plants carry out carbonic acid assimilation, it is necessary to supply a sufficient amount of carbon dioxide gas and a sufficient amount of light, and various methods have been proposed for preventing microorganisms from being mixed with the passage of these gases.

As the culture container, an Erlenmeyer flask having an air entry path smaller than the culture area is often used, and other test tubes, wide-mouthed bottles and the like are also used. Conventionally, a so-called aluminum cap has been used for these container mouths, which is covered with a cotton plug obtained by hardening cotton fibers and an aluminum foil, and the portion protruding from the container mouth is twisted by hand to close while forming wrinkles. This forms a passage between the outside air consisting of a winding bottleneck and the inside of the bottle in the periphery of the container mouth, and when air passes through this bottleneck, bacteria remain in the overflow channel and prevent contamination by bacteria.

In addition, screw lids and mating lids are also used, but in this case packing is not used, and when outside air circulates through the gap between the lid and the container that is inevitably left, the bacteria flow from bottom to top. Keeps aseptic because it is hard to move.

Even in tissue culture, since it is a plant, it photo-synthesizes and consumes a large amount of carbon dioxide. In the conventional method, the supply of carbon dioxide gas is insufficient, and when the illuminance increases, the carbon dioxide concentration decreases to 100 ppm or less, sufficient photosynthesis is not performed at such low concentration, and saccharose, glucose and other sugars must be added. did not become. Moreover, this seedling was very weak and had a low survival rate during acclimation.

Furthermore, since tissue culture seedlings of plants are probably handicraft and labor costs occupy most of the production cost, mechanization, labor saving, and rapid growth are required.
Japanese Patent Publication No. 4 discloses a technique in which the supply of water is ensured to allow sufficient air circulation and the working efficiency is improved.

[Problems to be Solved by the Invention]

However, there is a drawback that the medium is dried when the air flow is vigorously made and the carbon dioxide gas is sufficiently supplied. Tissue culture seedlings are usually grown in a sealed manner for 2 to 3 months, and during that time, it is necessary to prevent the culture medium from being dried to such an extent that the growth of plants is not adversely affected. Furthermore, when the amount of light is increased, the temperature inevitably rises, exceeds the optimum temperature for culturing the plant, and further causes the medium to dry, so that the expected result is not obtained.

Therefore, there has been a demand for means for preventing excessive temperature rise of the medium while performing sufficient irradiation.

[Means for solving problems]

The present invention is intended to solve the above problems, a part of which is a culture instrument comprising a water tank and a culture medium container in a plastic bag which is a microporous film that passes air and blocks microorganisms, It is characterized in that a light source is housed in the upper part of a culture unit housed so that water vapor evaporated from a water tank can reach the culture medium, a transparent plate is provided on the lower surface, and a light source chamber having a blower port and an exhaust port is provided. .

As the light source of the present invention, various light sources such as a fluorescent lamp and an incandescent lamp are widely used in addition to natural sunlight, but a fluorescent lamp which is generally used and has less heat generation compared to the amount of light is preferable. This light source is housed in a light source chamber whose lower surface is made of a transparent plate. In order to effectively utilize the light source, it is preferable that the inner surfaces other than the lower surface are mirror surfaces having reflectivity.

The transparent plate is usually glass, but may be a plastic plate, a plastic sheet or the like as long as it has high transparency. In general, UV light has a negative effect on biological tissues,
Since infrared rays are also heat rays, it is preferable to provide the transparent plate with an ultraviolet ray blocking property and an infrared ray blocking property. In order to impart such characteristics, UV or infrared blocking glass may be used, or a UV blocking or infrared blocking sheet may be attached to ordinary glass, or these sheets may be doubled. Alternatively, a paint having these characteristics may be applied.

Since the light source room becomes hot due to the light source, there is a ventilation port on one side.
On the other hand, an exhaust port is provided to remove the hot air inside. Therefore, since it does not reach the plant tissue other than the radiant heat, it is possible to prevent an excessive rise in the temperature of the medium even if a strong light source is used. The hot air discharged from the exhaust port can be effectively used by being used for drying various materials, products, preheating, and the like.

As a tissue culture unit for plants, a part is a microporous film that allows air to pass through and is a microporous film that blocks the culture equipment consisting of a water tank and a culture container, so that the water vapor evaporated from the water tank can reach the culture medium. A stored culture unit is preferred. The plastic bag is a transparent container made of heat-resistant glass, polypropylene, polycarbonate, polystyrene, high-density polyethylene, nylon, etc. that can withstand heat sterilization because it efficiently transmits the light required for photosynthesis and withstands repeated use, and the trunk and leaves grow. It is preferable that the portion above the culture medium to be used is transparent.

In addition, the medium and the water tank are separated, and in addition to the surface of the medium has a water surface separated from it, these are in a transparent plastic bag which is a microporous film that partially passes air and does not pass microorganisms Those stored in are preferable. As a means for separating, a two-stage structure is provided in which an upper box that serves as a culture medium is provided on a lower box that serves as a water tank, and a notch is provided at the upper end of the lower box.
There are methods such as forming a through hole above the water surface of the lower box, or providing a pipe-like through hole that projects from the bottom surface of the upper box onto the medium. Furthermore, a method of directly sucking the water in the lower box into the medium in the upper box by an assembly of capillaries such as strings or strings, or the water vapor in the lower box directly into the medium without dropping the medium or water on the bottom of the upper box It is also possible to provide a porous support plate that is absorbed. Further, the medium and the water tank may be arranged side by side.

It suffices that the water has an amount and an evaporating water surface sufficient to supply water evaporated from the medium during the culture period.

The culture medium container and the water tank are hermetically housed and sealed in a bag at the same time.
It is necessary that a part of this bag is made of a microporous film described later. Furthermore, it is preferable that the bag is transparent, and it is not necessary that the bag is entirely transparent as long as the above-mentioned plant grows from the culture medium in a part where the above-mentioned container grows.

The method of sealing the plastic bag may be any method as long as it can be kept completely airtight throughout the culture period, but the method of heat fusion is reliable and simple.

The microporous film used in the present invention has a pore size of 0.02μ or more, preferably 0.01μ or more microorganisms can not pass, if the difference between the major axis and the minor axis is small, the minor axis of the pores is 0. .
It is 1 μm or less, preferably 0.05 μm or less. Further, in the case of a slender elongated hole, if the maximum minor axis is 0.1 μm or less, preferably 0.05 μm or less, more preferably 0.02 μm or less, passage of microorganisms can be prevented even if the major axis is large. In short, a film having pores as small as possible within a range in which oxygen and carbon dioxide can flow without resistance may be used.

A monolayer film is preferable in order to reduce the flow resistance of gas and to supply carbon dioxide more smoothly. In general, many microporous films have layers having large pores that are overlapped with each other to form a bottleneck. In such a microporous film having a multi-layered structure, the ventilation resistance is inevitably increased, which tends to hinder the smooth supply of carbon dioxide gas.
Moreover, microorganisms having no self-mobility cannot pass through the bottleneck and are blocked, but invasion of microorganisms having self-mobility such as fine mites cannot be completely prevented.

The microporous film material is preferably a water repellent film such as polypropylene, polyethylene, nylon, and fluorine resin. Since microorganisms mainly move with water, even if the pores have the same size, it is difficult for the microorganisms to pass if the microporous film material is water repellent. In addition, the drying of the medium is due to the evaporation of water contained in the medium, but in the case of a water-repellent microporous film, hydrophobic carbon dioxide and oxygen pass well but water vapor does not pass easily, It has a good air-permeability but also has some effect of preventing the medium from drying.

To use a microporous film as a part of a plastic bag, for example, a part of the bag, preferably a side part when a container is stored, is cut at one or more places to form a through hole, and a non-woven fabric or the like is provided in the through hole. The reinforced microporous film may be heat-sealed. Alternatively, holes may be intermittently provided in the bag, and the reinforced microporous film may be heat-sealed or adhered to cover the holes.

When the culture medium container and the water tank are not hermetically housed in the bag, the culture medium container and the water tank are hermetically sealed with a lid, and the microporous film is attached to the lid. Further, it is necessary that the upper space of the culture medium and the upper space of the water tank are in communication with each other. When the culture medium container and the water tank are stacked vertically in two stages, the upper space of the ice bath of the lower box and the culture medium may communicate with each other.

[Action]

According to the present invention, even when strong light is irradiated from the upper part of the tissue culture tank, the light source chamber is shielded from the culture unit by the transparent plate, and the heated air is separately removed, so that the temperature of the medium increases. It is possible to grow a strong seedling in a short period of time by irradiating sufficient light and supplying sufficient air without causing any harmful effects, and it can be directly planted as it is without acclimation.

〔Example〕

FIG. 1 is a perspective view of one embodiment of the present invention, and FIG. 2 is a sectional view. Reference numeral 1 is a shelf, and 2 is a leg portion that supports the shelf. Three
Is a light source chamber, the lower surface of the shelf 1 is used as the upper surface of the light source chamber, and three fluorescent lamps 4 are arranged inside. 5 is a connector.
Reference numeral 6 denotes a glass plate provided on the lower surface of the fluorescent lamp 4 and forms the lower surface of the light source chamber 3. The fluorescent lamp 4 is hermetically sealed by the glass plate 6, the lower surface of the shelf 1 and the side end portion 9 except the intake port 7 and the exhaust port 8. An ultraviolet blocking film and an infrared blocking film were attached to the glass plate 6. Reference numeral 10 denotes a motor provided in the intake port 7. The intake fan 11 driven by the motor 10 rotates to forcibly suck the air into the light source chamber 3, and the hot air heated by the fluorescent lamp is discharged from the exhaust port 8. FIG. 2 shows a state in which the exhaust duct 8 is provided at the exhaust port 8. Reference numeral 12 is a motor support bracket. The culture unit 13 is arranged on the shelf 1. The culture unit is irradiated with sufficient light, and the temperature does not rise excessively.

The exhaust port 8 may be simply opened as shown in FIG. 1, but in the case of culturing in a large amount, an exhaust duct 22 is provided as shown in FIG. Conduct.

In this embodiment, a culture tank having a ventilation hole 16 is also provided above the water tank 15 having a ventilation hole 14 as the culture unit 13.
17 bags are placed and a bag made entirely of polypropylene film
Sealed to 18. A hole was made in the bag 18, and air was passed through the hole, and Juraguard (trademark) manufactured by US Celanese Co., Ltd. was fused as a microporous film 19 for blocking microorganisms.
20 is a medium and 21 is water. In this way, the culture tank 17 is maintained in a sterile state while being supplied with sufficient air. Further, since the culture medium tends to be dried because the air flow from the microporous film 19 is free, the water in the water tank 15 evaporates, the bag 18 is filled with the air from the ventilation hole 14, and the culture is performed from the ventilation hole 16. Enter the tank 17. Therefore, in this example, it was possible to prevent the culture medium from drying even if the air flow was free.

In the present embodiment, the motors are provided in the light source chambers 3 of the respective stages, but it is also possible to blow air to the intake ports of the plurality of light source chambers 3 in which one motor is provided in the collective intake pipe. Alternatively, the exhaust port 8 of one light source chamber 3 may be connected to the intake port 7 of the adjacent light source chamber.

〔effect〕

According to the present invention, the temperature of the medium does not rise abnormally even when irradiated with strong light, the medium does not dry, and the plantlets can be robustly grown by receiving sufficient light. The tissue-cultured seedlings obtained by the present invention are strong and do not need acclimation, and can be directly planted as they are, thus expanding the types of plants that can use the tissue-cultured seedlings.

[Brief description of drawings]

The drawings show an embodiment of the present invention. FIG. 1 is a perspective view and FIG. 2 is a sectional view. In the drawings, reference numeral 1 is a shelf, 2 is a leg portion, 3 is a light source chamber, 4 is a fluorescent lamp, 5 is a connector, 6 is a glass plate, 7 is an intake port, 8 is an exhaust port, 10 is a motor, and 11 is an intake fan. , 13 is a culture unit, 14 and 16 are ventilation holes, 15 is a water tank, 17 is a culture tank, 18 is a bag, 19 is a microporous film, 20 is a culture medium, 21 is water, and 22 is an exhaust duct.

Claims (2)

[Claims] 1. A culture bag comprising a water tank and a culture medium container in a plastic bag, a part of which is a microporous film that allows air to pass through and blocks microorganisms, so that water vapor evaporated from the water tank can reach the culture medium. At the top of the stored culture unit,
A plant cultivating device, which houses a light source, has a transparent plate on a lower surface, and is provided with a light source chamber having a ventilation port and an exhaust port. 2. The plant culture apparatus according to claim 1, wherein the culture device has a culture medium container placed on a water tank, and a notch or perforation is provided on the upper portion of the water tank.