JP5821427B2 - Biological treatment apparatus, gene processing apparatus, and biological breeding system including the same - Google Patents

Description

  The present invention relates to an organism processing apparatus, and more particularly to a gene processing apparatus. Moreover, it is related with the organism training system provided with them.

  In order to confer resistance to organisms such as plants, in order to create faster-growing organisms, and because petals have a more vivid hue, attempts to transform plants have been widely conducted. Yes. A vacuum infiltration method is known as a method used when transforming such an organism (for example, Non-Patent Document 1).

  The vacuum infiltration treatment is a typical method for producing a genetically modified plant individual, which is simpler and quicker than other methods such as tissue culture methods using explants such as roots and stems, and the skill of the operator. It has the distinct advantage of not requiring it and is commonly used. The simple operation method will be described below.

  First, a healthy plant is grown, and the above-ground part of the plant body is immersed in a suspension for a recombination composition to be introduced and placed under reduced pressure so as to infiltrate the suspension into the plant body. Examples of the composition for recombination include a composition containing Agrobacterium for gene recombination. In the case of using Agrobacterium, after the above-described decompression infiltration operation, infection of Agrobacterium to the plant body and transfer of the T-DNA site, which is a part of the plasmid gene possessed by Agrobacterium, to the plant body are performed. However, when the transfer occurs in the flower, the T-DNA is transmitted to the next generation seed through fruiting, and the transformed first generation is contained in a certain ratio in the seed. This is a so-called genetically modified plant.

"Model Plant Lab Manual", 2000 Masaki Iwabuchi and others Springer Fairlark Tokyo

In the vacuum moistening method in genetic recombination, after removing the plant-flowering flowers, the plant is suspended in the pot while the plant is planted in the pot with respect to the dispersed Agrobacterium suspension. Genetic recombination is performed by dipping in a suspension and reducing the atmosphere (see Non-Patent Document 1).
And the operation | work which inverts a bowl in this way and soaks in a suspension or takes out is mainly performed manually. When such gene recombination is performed manually, it is very inefficient, and when the plant is moved or immersed in the suspension, the plant is stressed and leaves and stems break, There was a fear of entering.
In addition, the gene recombination treatment often requires a decompression treatment, and there is a problem that it is difficult to maintain airtightness in a facility capable of gene recombination.
This invention solves such a problem, and makes it a subject to provide the biological treatment apparatus suitable for biological treatment.

The present inventors have examined a biological treatment apparatus capable of efficiently performing genetic recombination operations while ensuring airtightness. As a result, a fixing member for holding and fixing the biological body in the biological treatment apparatus having airtightness is provided. The present inventors have found that the above problem can be solved by enabling reversal, and have completed the present invention.
The present invention includes a casing having an opening for introducing and discharging organisms, a lid that can seal the opening of the casing, a fixing member that is located inside the casing and holds and fixes the organism, and a liquid in the casing A biological treatment apparatus comprising supply / discharge means for supplying and discharging
The casing has airtightness by sealing the opening with a lid,
The fixing member is a biological treatment apparatus that can be reversed while holding and fixing the biological matter.

  Another aspect of the present invention is a biological treatment method performed using the biological treatment apparatus, wherein the biological member is brought into contact with the liquid in the casing by reversing the fixing member. A biological treatment method comprising:

In another aspect of the present invention, a casing having an opening for introducing and discharging organisms, a lid that can seal the opening of the casing, a fixing member that is located inside the casing and holds and fixes the organisms, And a gene introduction processing device comprising supply and discharge means for supplying and discharging liquid in the casing,
The fixing member is a gene introduction processing apparatus that can be inverted while holding and fixing an organism, and that performs gene introduction processing by inverting the fixing member.

Another aspect of the present invention is an organism training system in which an organism training apparatus having a space capable of growing an organism is connected to the organism processing apparatus or the gene transfer processing apparatus,
The living body breeding system includes a transporting unit that can transport the living body between the living body growing space and the living body processing apparatus or the gene introduction processing apparatus.

The organism treatment apparatus of the present invention eliminates the need for manual transfer and inversion of organisms, reduces stress on the organisms, and enables genetic recombination processing efficiently. In addition, since the device has airtightness, contamination of germs and the like can be prevented.
In addition, since the organism growing apparatus is connected to the organism processing apparatus and the organism growing system is provided with transfer means, the organism can be easily transferred between the organism processing apparatus and the organism growing apparatus. In addition, automation of the entire system can be easily achieved.

It is a schematic diagram showing the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the inversion state of the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the external appearance of the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the inversion state of the one aspect | mode of the biological treatment apparatus of this invention. It is a schematic diagram showing the one aspect | mode of the biological body cultivation system of this invention. It is a schematic diagram showing the one aspect | mode of the biological body cultivation system of this invention.

  An object processing apparatus of the present invention includes a casing having an opening for introducing and discharging a living organism, a lid capable of sealing the opening of the casing, a fixing member that is located inside the casing and holds and fixes the living organism, and It is a biological treatment apparatus provided with the supply / discharge means which supplies / discharges the liquid in the said casing.

  A casing introduces a biological body into it and processes a biological body. The shape of the casing is not particularly limited, but is preferably a cylindrical body or a columnar body from the viewpoint of ease of reversal operation, ease of fixing, ease of introduction and discharge of organisms, and the like. A shape having a same cross section of a point target shape such as a circular shape, an elliptical shape, a square shape, a rectangular shape, a hexagonal shape, and an octagonal shape is preferable. Further, in order to accommodate a reasonable and efficient quantity of organisms, when the casing is cylindrical or columnar, it is preferable that the casing is long in the longitudinal direction.

  Depending on the type of biological treatment, the treatment may be performed under reduced pressure. In order to cope with such a case, the casing is preferably made of a material that can withstand a reduced pressure state, and may have a reinforcing structure. As the reinforcing structure, the strength of the casing can be improved by arranging a reinforcing material around the casing and fixing the casing and the reinforcing material by bonding or welding. The reinforcing material is preferably one having high strength, but is not particularly limited, and examples thereof include metal materials, plastic materials, and wooden materials. It is also possible to use the same material as the casing as the reinforcing material.

  The material of the casing is not particularly limited as long as it is a material that can be used for normal biological treatment. In addition, when the organism is treated, airtightness to the extent that various germs and the like are not mixed is required. In terms of moisture resistance and strength, a metal material having corrosion resistance is preferable. Examples of the metal material include stainless steel, hastelloy, rust-proof carbon steel, aluminum, zinc alloy, brass, copper, bronze, and their materials. An alloy etc. are mentioned. Of these, stainless steel is preferable in terms of economy. Further, in the biological treatment apparatus of the present invention, when the biological material is grown using an artificial light source, at least a part of the casing material is preferably transparent. Specific examples of the transparent material include acrylic plastic, transparent plastic materials such as polystyrene and polycarbonate, and glass.

  The size of the casing is not particularly limited, and can be set as appropriate in consideration of the size and amount of the organism to be treated. Since it is necessary to have a minimum volume for accommodating a living organism, the cross section is usually 10 cm × 10 cm or more, preferably 20 cm × 20 cm or more, more preferably 30 cm × 30 cm or more, and in the longitudinal direction of the casing. The length is usually 20 cm or more, preferably 50 cm or more, more preferably 70 cm or more. In addition, from the viewpoint of throughput of decompression and decompression operations, and from the viewpoint of the case where it is necessary to have strength that can withstand decompression, the cross section of the casing is usually 3 m × 3 m or less, preferably 1 m × 1 m or less, more preferably Is 70 cm × 70 cm or less, and the length of the casing in the longitudinal direction is usually 10 m or less, preferably 5 m or less, more preferably 2 m or less.

The casing has an opening for introducing and discharging organisms. The number, position, and size of the opening are not limited as long as one or more openings are provided, but it is preferable that the opening is located on the side surface of the casing in the longitudinal direction. It is more preferable to have two openings and to face the side surface of the end of the casing in the longitudinal direction.
The size of the opening is not particularly limited, and the size capable of introducing and discharging organisms can be appropriately selected. When located on the side surface of the longitudinal end portion of the casing, it may be the same size as the cross section perpendicular to the longitudinal direction of the casing, or may be slightly smaller.

When the biological treatment apparatus of the present invention is incorporated into a biological growth system, the biological growth apparatus is connected to one or both of the longitudinal directions of the casing. The opening of the casing in this case is provided with an opening at one or both of its longitudinal ends in accordance with the arrangement of the organism growing device. In the case of such an aspect, in order to facilitate introduction / discharge of the organism from the organism growth apparatus to the organism treatment apparatus, the same as the opening in the organism growth apparatus on the side surface in the longitudinal direction of the casing. It is preferable that the opening has a size because the casing and the organism growing apparatus can be smoothly connected. Moreover, in order to fix a casing and a biological body growing apparatus, it is preferable to equip both with a flange structure and to fix in a flange. As a fixing means, fixing by bolts & nuts or clamps is preferable.

  The shape, size, material, and the like of the lid that can seal the opening of the casing are not particularly limited, and can be appropriately selected according to the shape and size of the opening of the casing. Also, the material is not limited as long as it can seal the opening of the casing and has airtightness for performing biological treatment, and the same material as the casing can be used. Specifically, it is preferable to use stainless steel having high strength and low risk of corrosion due to moisture.

For sealing the casing and the lid, it is preferable that both be provided with a flange structure and fixed at the flange. As a fixing means, fixing by bolts & nuts or clamps is preferable.
In addition, depending on the type of biological treatment, the treatment may be performed under reduced pressure. In order to cope with such a case, a reinforcing plate made of a metal material or the like may be provided together in fixing the casing and the lid.

By sealing the opening of the casing with a lid, the inside of the casing of the biological treatment apparatus of the present invention has airtightness. The biological treatment apparatus of the present invention preferably has airtightness when a negative pressure is applied to the inside of the container in order to effectively infiltrate the liquid into the biological body, for example, in the treatment by the vacuum wetting method. That is, it is preferable to have airtightness between the casing and the lid so that outside air does not enter during the decompression process.
In this way, when the casing is under reduced pressure, when the organism placed inside is a plant, air is sucked out mainly from the space between the cells inside the leaves of the plant during negative pressure. When the negative pressure is restored, the suspension is physically injected into the space where the intercellular spaces exist, and the plant can be subjected to the desired treatment. When the target treatment is recombinant gene transfer using Agrobacterium, Agrobacterium is sucked into the leaves of the plant together with the suspension when the pressure is restored, and the gene transfer is preferably performed.

  The airtightness of the biological treatment apparatus of the present invention is usually sufficient to have airtightness at normal pressure, but depending on the type of biological treatment, it is necessary to have airtightness even under reduced pressure. Usually, it is sufficient to ensure airtightness at 1 atm (760 Torr), but it is preferable to ensure airtightness at 100 Torr for 10 minutes, more preferably at 25 Torr for 10 minutes.

  In the biological treatment apparatus of the present invention, when the genetic recombination treatment is performed by the wet decompression method, a decompression device necessary for forcibly exhausting the air inside the casing is provided to decompress the inside of the casing. It is done. The decompression device is not particularly limited as long as the inside of the casing can be decompressed. However, it is preferable that the casing includes an exhaust port, and the exhaust port is connected to the decompression device. The exhaust port may be connected to the decompression device by a tube or piping. Examples of the material of the tube include resins such as rubber, silicone, and tetrafluoroethylene. Examples of the piping material include metals such as carbon steel, copper and brass, and resins such as vinyl chloride. Examples of the decompression device include a vacuum pump and an aspirator.

Further, in order to restore the pressure inside the decompressed casing, a leak port necessary for introducing air into the casing is provided. The leak port may be provided separately from the exhaust port, or the leak port and the exhaust port may be the same member, and the tube or pipe connected to the exhaust port and the pressure reducing device may be branched at a halfway point. By providing a valve, the pressure reducing function and the returning pressure function may be provided. Moreover, in order to confirm the pressure reduction degree inside a casing, you may provide the pressure gauge.

In order to prevent moisture and volatile components inside the casing from excessively entering the decompression device, a mist trap having a function of collecting liquid mist may be interposed in a tube or piping connecting the casing and the decompression device. Good.
For the purpose of observing the inside of the casing in the process of decompressing the inside of the casing, an observation window may be provided in the transparent casing. The shape of the observation window is not limited as long as the airtightness in the installation portion is not impaired. As the material of the transparent portion, glass or a transparent resin can be usually used.

The fixing member of the present invention is located in the casing and holds and fixes the organism. In the biological treatment apparatus of the present invention, the fixing member can be reversed while holding and fixing the biological body. Therefore, it is necessary to fix the biological body so that the biological body is not detached or dropped during the reversal.
As described above, a known structure and means can be used as means for holding and fixing the organism to the fixing member when the fixing member is reversed. Specific examples are given below.

Examples of the fixing member include a member having a concave portion having an appropriate size and shape that can accommodate a root portion of a living organism. The material of the member is not limited, and examples thereof include resins such as vinyl chloride, wood, bricks, and metals. It is preferable that it consists of resin from a viewpoint of weight reduction or ease of handling. It is sufficient that at least one recess is present, but the number may be appropriately set according to the efficiency of processing by the apparatus. Moreover, the member for fixing a biological body to a recessed part is also required. For example, the root part of a living organism is sealed with a sealing material such as urethane resin, rock wool, or an inorganic porous material, and the sealing material and the fixing member are fixed by means such as adhesion. In addition, the soil may be fixed by pouring soil into the recess with the root portion of the organism arranged in the recess and closing the recess with a sealing member such as a plastic plate. In addition, a concave portion of the fixing member and a detachable cup are separately prepared, and the organism is disposed and fixed in the cup, and the cup is attached to the concave portion of the fixing member, so that the organism for biological treatment It is preferable because exchange can be performed smoothly and without stressing the organism.
In addition, the fixing member is preferably detachable from the casing. In this case, since the whole fixed member can be taken out from the casing and the organism can be exchanged, the organism can be exchanged without applying stress.

  The fixing member may or may not be fixed to the casing. In the case of being fixed, the fixing means is not particularly limited. When the casing is cylindrical, the end of the fixing member and the peripheral edge of the casing may be fixed by the coupling means, and the end of the fixing member and the side surface of the longitudinal end of the casing are fixed by the coupling means. Also good. Moreover, a transfer rail may be arrange | positioned to a casing and a fixing member may be fixed on a transfer rail. When such a transfer rail is arranged, it is possible to smoothly transfer the fixing member to which the organism is fixed into the casing, which is preferable. When the fixing member and the casing are fixed, the fixing member also follows and reverses by reversing the casing.

  The fixing member of the biological treatment apparatus of the present invention is characterized in that it can be reversed. As a means for reversing the whole casing, for example, a guide roller and a turning roller are installed in the lower part of the casing, and the reversal is performed by driving the turning roller. In the case of such an embodiment, the casing is preferably a cylinder from the viewpoint of easy reversal. The power for reversing may be electric power such as an electric motor, or can be reversed manually. In the case of manual operation, it is possible to reverse the casing by a reversing operation of a lever or the like using a device having a speed increasing / power increasing mechanism using a gear or the like.

The contact surface between the casing and the turning roller is preferably subjected to anti-slip processing such as rubber, gears, and turret processing. By setting it as such an aspect, reversing operation can be performed stably and adhesion to the unnecessary part of the biological body treatment liquid by having reversed too much can be prevented. Moreover, the wall for fixing a casing and the pillar are arrange | positioned in the outer side surface part of a casing, and the installation of the roller for inverting such a fixing member becomes easy.

  On the other hand, when the fixing member and the casing are not fixed, the fixing member is fixed to the reversing shaft extending in the longitudinal direction of the casing, and the reversing shaft is penetrated to the side surface of the longitudinal end portion of the casing. It can also be reversed. In addition, a guide ring that is not fixed to the casing but fixed to the fixing member and can be reversed together with the fixing member can be provided inside or outside the casing. When the fixing member and the casing are not fixed as described above, the fixing member is inverted by inverting the reversing shaft or the guide ring without inverting the casing itself.

  In the case of such an aspect, a motor for reversing the reversing shaft may be installed, or a turning roller for reversing the guide ring may be separately provided. It is also possible to manually reverse the reversing shaft or reverse the guide ring.

  The means for supplying and discharging the liquid into the casing is not particularly limited as long as it is a means capable of supplying the liquid necessary for the treatment of the organism into the casing and discharging it from the casing. As a specific example of the means for supplying and discharging the liquid into the casing, supplying and discharging the liquid through a hole or the like can be mentioned. In the case of using a hole, the position of the hole is not particularly limited, and can be set as appropriate in consideration of the reversible fixing means.

If the casing is also reversed when the reversible fixing means is reversed, it is sufficient to install only one hole. Usually, when supplying the liquid, it is only necessary to be positioned above the central portion of the casing. In such a configuration, while the liquid is reversed and the treatment of the organism is performed, a hole is formed by a stopper or the like. It is necessary to close it.
On the other hand, when the reversible fixing means is reversed, when the casing does not follow and the fixing means alone is reversed, it is preferable to arrange the holes above and below the central portion of the casing. In this case, the hole arranged below the central portion must be closed with a plug or the like when filling the liquid, and the liquid can be removed by removing the plug when discharging the liquid. Can be discharged.

  The size of the hole is not particularly limited, and can be appropriately set in consideration of the size of the casing. Generally, it is about φ5-30 mm. In addition, when a genetically modified product is contained in the liquid, it is necessary to inactivate the gene by autoclaving or formalin treatment in order to prevent its diffusion to the external environment. A processing apparatus for inactivation can be provided separately. After the gene is inactivated, it can be discarded by the same method as that for normal industrial waste liquid treatment.

The biological body processing apparatus of this invention is an apparatus for performing the process to a biological body. The organism is not particularly limited as long as it can be treated with a liquid, but is preferably a plant.
Although it does not specifically limit as a plant, It applies preferably to the angiosperm plant generally used as a plant for gene recombination, or the plant which attaches a leaf with a comparatively big area. Specific examples of such plants include: Gramineae, Gramineae, Wheat genus, Sugarcane genus, etc., Gramineae, Duckweed, etc. Legumes such as genus, pea genus, and genus Leguminosae, eggplant genus, capsicum genus, tobacco genus, etc. Aceraceae, cotton genus, etc., mallow, mushrooms, apples, etc., roses, cassaceae, etc. And plants of the family Brassicaceae such as Wasabi spp.

Moreover, it is preferable to use the biological body processing apparatus of this invention for the process to a hydroponics plant.
Hydroponic cultivation does not use a medium that easily contains germs and the like, and therefore can be cultivated in a considerably aseptic condition. Since this processing apparatus can reduce the opportunity to touch outside air in the operation process, it is advantageous in that it does not easily impair the features of aseptic cultivation.
Hydroponics is often cultivated with the underground part exposed in the cultivation nutrient solution. Therefore, when the plant body is physically manipulated, the underground part may be damaged, or excessive stress may be applied to the underground part. In particular, operations that invert the top and bottom, such as the treatment in the vacuum moistening method, may cause excessive stress on the plant body, adversely affect growth, or cause unwanted secondary effects. There has been a problem with increasing the possibility of side effects in which metabolites are produced. However, if the processing apparatus of this invention is used, it will become possible to minimize a physical load to the underground part of a plant body, and load stress can be reduced extremely. In addition, since the reversal operation is rational, it is extremely advantageous from the viewpoint of productivity.

Processing by the biological treatment apparatus of the present invention is not particularly limited as long as it can be performed by the apparatus of the present invention. Preferably, it is the process which makes a liquid contact the leaf stem part of a plant.
The treatment of bringing the liquid into contact with the leaf stem part of the plant is divided into a method of immersing the liquid in the leaf stem part of the plant under reduced pressure and a method of immersing the liquid in the leaf stem part of the plant under normal pressure. It is done. As a method for immersing the liquid in the leaf stem portion of the plant under reduced pressure, there is a reduced pressure wetting method (Infiltration).
As a specific step of the reduced pressure wetting method, first, a plant body is grown to a certain extent, and the above-ground part, particularly the leaf part, of the plant body is immersed in a liquid prepared according to the purpose of treatment and decompressed. In this step, air is removed from the portion corresponding to the gap between the cell tissues constituting the above-ground part of the plant body. Next, the process of releasing the decompression and restoring the pressure is performed. Here, due to the restoring force of the cell wall constituting the gap of the cell tissue, a negative pressure is generated in the area where the gap exists, and the suspension is formed in the space. Physically infiltrate. When the liquid is a suspension composition containing Agrobacterium for genetic recombination, Agrobacterium can be infiltrated into the portion where the gap exists at the time of infiltration.

On the other hand, as a method for immersing the liquid in the leaf stem portion of the plant under normal pressure, there is a Filar dip method.
The Filoral dip method is a method similar to the Infiltration method, except that the decompression and decompression operations are not performed. That is, it is a method of immersing the suspension in the plant body under atmospheric pressure.

When the biological treatment by the biological treatment apparatus of the present invention is the above-described reduced pressure wetting method, which is a gene introduction treatment, the conditions are not particularly limited, and the conditions such as temperature, pressure, etc., under which a normal reduced pressure wetting method is performed may be carried out. it can.
When the pressure is reduced to the desired degree of pressure reduction, it is preferable that the pressure is gradually or stepwise reduced rather than abruptly reduced from the atmospheric pressure. For example, a speed of about 50 to 300 Torr / min is usually used.
Similarly, when returning from a predetermined degree of pressure reduction to return to atmospheric pressure, it is preferable to return the pressure gradually or stepwise.

Further, the degree of pressure reduction in the negative pressure operation is not limited as long as the purpose is achieved, but is usually 500 Torr or less and 20 Torr or more. When the suspension is an aqueous solution, the degree of vacuum is preferably such that water does not reach the boiling point. When the temperature is 25 ° C., the preferred degree of vacuum is about 25 Torr. When the pressure of decompression is too high, liquid infiltration becomes insufficient, which is not preferable. On the other hand, when the pressure is too low, the liquid reaches the boiling point, remarkably evaporates, the liquid is reduced, and the infiltration becomes insufficient.

  In the biological treatment apparatus of the present invention, when handling a substance or a living body for the purpose of genetic recombination, in order to prevent the recombinant gene from adversely affecting the human body, the environment, etc. It is necessary to provide a structure. Before the gas exhausted from the inside of the container is released to the outside world, the genetically modified product is collected by an installed filter or the like. You may equip the equipment for performing a gene inactivation process as needed.

  The organism treatment apparatus of the present invention is preferably used as an organism cultivation system by linking and arranging an organism cultivation device having a space capable of growing an organism. By setting it as such a biological body cultivation system, a biological body can be raised efficiently.

  It is preferable that the organism growing apparatus which has the space which can grow an organism which comprises an organism growing system is used for hydroponics. When using it for hydroponics, it sets so that it may become an environment suitable for hydroponics of a plant in the living body upbringing apparatus. Usually, the temperature is 15 to 35 ° C., the humidity is 30 to 95% RH, and an air stream is preferable because photosynthesis is promoted. It is also preferable to artificially introduce carbon dioxide because photosynthesis is promoted. When carbon dioxide is artificially introduced, the concentration is usually 400 to 2000 ppm.

  In the case of hydroponics, a so-called nutrient solution that is an aqueous solution in which a nutrient component is dissolved is brought into contact with the root of the plant body. It is preferable to provide a mechanism for keeping the concentration of nutrient components in the nutrient solution constant. Nutritional components are usually normal fertilizer components such as phosphorus, nitrogen, potassium and the like. The concentration can be managed, for example, by a method of managing the ion concentration by measuring electrical conductivity or the like.

  Although the shape of the organism growing apparatus is not particularly limited, it is possible to accommodate the organism and preferably keep the temperature in the apparatus constant, and it is preferable to have a cylindrical shape. In addition, when the organism growing apparatus has a cylindrical shape, one end thereof is connected to the organism processing apparatus of the present invention, but the opening at the end is connected to the organism processing apparatus of the present invention for ease of connection. The shape is preferably substantially the same as the opening of the device.

  In addition, the living body growing device preferably has an artificial light source in order to increase the efficiency of photosynthesis. The artificial light source is not particularly limited as long as it is a light source capable of photosynthesis, and examples thereof include a fluorescent lamp, a cold cathode tube, a sodium lamp, a mercury lamp, a light emitting diode, and electroluminescence.

  Moreover, in the biological body cultivation system of this invention, it can transfer to a biological body processing apparatus from a biological body cultivation apparatus, and can grow and process continuously. In this case, transfer can be performed smoothly by connecting the organism growing apparatus and the organism processing apparatus in the horizontal direction. Although the specific method of transfer is not particularly limited, the member to which the organism is fixed can be moved in and out in the horizontal direction by a method such as sliding on the guide rail or moving by hanging on the transfer guide. A guide roller may be provided on the guide rail for the purpose of facilitating transfer.

  When a liquid is infiltrated into an organism for the purpose of gene introduction, the organism is stored in a static environment or in an appropriate environment after treatment to express the gene. When the organism is a plant, it is placed under appropriate light, temperature and humidity conditions so that the gene is appropriately expressed.

  Hereinafter, specific embodiments of the present invention will be described with reference to the drawings. However, the present invention is not limited to the specific embodiments shown in the drawings.

  FIG. 1 is a diagram showing an embodiment of the biological treatment apparatus of the present invention. The casing 1 has an opening, and the organism 2 is introduced and discharged from the opening. The shape of the casing 1 is not particularly limited as long as the organism 2 can be accommodated inside the casing. The biological treatment apparatus of FIG. 1 is rectangular, but it may be cylindrical as shown in FIG. The opening of the casing 1 is provided with a lid 3, and the casing 1 can be sealed by closing the lid 3 in the direction of the arrow, and the casing 1 has airtightness.

  As for the airtightness of the casing 1, it is sufficient to block outside air from outside at normal pressure. However, when performing genetic recombination treatment, it is preferable to have airtightness under reduced pressure, and airtightness for 10 minutes at 25 Torr. It is preferable that the security can be secured. Therefore, it is preferable to use a material that can withstand such a reduced pressure as the material of the casing 1.

  The organism 2 is held and fixed to the fixing member 4, and the fixing member 4 is fixed to the casing 1 via the coupling members 5 and 6. Further, the casing 1 includes a supply port 7 for supplying the liquid into the casing and a discharge port 8 for discharging the liquid out of the casing. Moreover, the casing 1 is provided with the exhaust port 9, and the exhaust port 9 is connected with the decompression device 10 through the pipe | tube. Further, the pipe connecting the exhaust port 9 and the decompression device 10 is branched in the middle, and one end thereof is a leak port 91. After the casing 1 is decompressed in the biological treatment, the inside of the casing can be decompressed by opening the leak port 91.

The biological treatment apparatus of the present invention is characterized in that the fixing member 4 is reversed while holding and fixing the biological body 2. Examples of invertible structures are shown in FIGS.
FIG. 2 is a cross-sectional view of a surface perpendicular to the longitudinal direction of a rectangular casing 1 in the biological treatment apparatus of the present invention. The casing 1 is fixed at both ends with a fixing member 4 to which the organism 2 is fixed. The casing 1 is also fixed with the guide roller 11, and the guide roller 11 is reversed by driving of the turning roller 12, and accordingly, the casing 1 is also reversed, and the fixing member 4 fixed to the casing 1 is also reversed. . An inverted diagram of the inverted situation is shown on the right. In the reverse view, a state in which the liquid is filled is shown, and the liquid level 13 is filled so as to immerse the leaf stem portion 21 of the organism 2 that is a plant.

  Moreover, FIG. 3 is the schematic which looked at the biological treatment apparatus from the exterior about arrangement | positioning of the guide roller 11, the turning roller 12, and the casing 1 of the biological treatment apparatus demonstrated in FIG. The turning roller 12 is driven manually or automatically, and the organism can be reversed when the guide roller 11 and the casing 1 are reversed.

In FIG. 4, the schematic of the biological body processing apparatus of this invention in case the casing 1 is cylindrical shape is shown. The cylindrical casing 1 is inverted in a state where the fixing member 4 to which the organism 2 is fixed is accommodated, but the organism 2 and the fixing member are inverted by inverting the two inversion shafts 14 fixed to the fixing member 4. 4 can be inverted. At this time, when the fixing member 4 and the casing 1 are fixed, the casing 1 is also inverted as the fixing member 4 is inverted. When the fixing member 4 and the casing 1 are not fixed, only the fixing member 4 is inverted. Will be reversed.
Although there are two inversion axes 14 in the figure, it is preferable to use a plurality of inversion axes from the viewpoint of stability during inversion. However, it may be reversed by one reversing shaft, and in that case, it can be easily fixed to the fixing member 4 by making the cross-sectional shape of the reversing shaft rectangular.

In FIG. 4, the reversal according to an aspect in which the longitudinal direction of the reversing shaft 14 is parallel (substantially parallel) to the longitudinal direction of the casing 1 is illustrated, but in FIG. 5, the longitudinal direction of the reversing shaft is the longitudinal direction of the casing 1. It shows a mode that goes straight. The reversing shaft 14 can be arranged as described above, and the fixing member can be reversed.

  FIG. 6 is a mode in which a circular guide ring 15 is provided on the outer side of the cylindrical casing 1, a turning roller 12 is disposed between the casing 1 and the guide ring 15, and the inner casing 1 is inverted. Even in such an embodiment, the organism can be easily inverted.

FIG. 7 is a schematic view of the organism training system of the present invention. By connecting the organism growing apparatus 18 having the organism growing space to both ends in the longitudinal direction of the casing according to the organism processing apparatus of the present invention, the organism growing system of the present invention is obtained.
Although not shown in the figure, a flow of liquid fertilizer is formed in the lower part of the organism growing apparatus 18 in the longitudinal direction of the organism growing apparatus, and the root part of the organism is immersed in the liquid fertilizer to the organism. Supply and nurture nutrients. Moreover, the liquid fertilizer which flows into the living body growing space can be circulated in the apparatus by providing the liquid circulating apparatus outside the living body growing apparatus 18. At this time, by controlling and adjusting the concentration of the liquid fertilizer, it becomes possible to make the liquid fertilizer concentration more appropriate for the growth of the organism.

  It is preferable that the organism growing system includes a transfer means including a transfer roller 16 and a transfer rail 17 so that the fixing member 4 to which the organism is fixed can transfer the organism growing device and the organism processing apparatus. According to such an embodiment, it is not necessary to manually move the organism, and it is difficult to apply excessive stress to the organism.

As the transfer means, as shown in FIG. 8, it is possible to hang and fix a fixing member to which a living organism is fixed to the transfer guide 20. Even in such an embodiment, the plant body can be smoothly transferred.
In addition, it is preferable that the organism training system includes an artificial light source 19. The artificial light source allows the plant body to perform photosynthesis regardless of day and night, and the growth of the plant body is accelerated.

The organism treatment apparatus of the present invention can be used for the treatment of organisms in general, and is particularly suitably used for gene recombination treatment such as a vacuum wetting method. Use of this device is expected to improve the growth rate of organisms.
In addition, by combining with a living body growing apparatus to form a living body growing system, it is possible to improve the growing speed of the living body and to automate a series of flows from growing to genetic recombination processing.

1 Casing 2 Plant (organism)
21 Plant body leaf stem portion 22 Plant body root portion 3 Lid 4 Fixing member 5, 6 Coupling member 7 Liquid supply port 8 Liquid discharge port 9 Exhaust port 91 Leak port 10 Decompression device 11 Guide roller 12 Turning roller 13 Liquid surface 14 Reverse shaft 15 Guide ring 16 Transfer roller 17 Transfer rail 18 Organism growing device 19 Artificial lighting 20 Transfer guide

Claims (17)

Supplying and discharging casing, can lid sealing the opening of the casing, the casing interior located fixing member for holding and fixing the organism, the liquid prior Symbol casing having an opening for introducing and discharging the organism A biological treatment apparatus comprising supply / discharge means and a decompression device capable of reducing the pressure inside the casing ,
The casing has airtightness by sealing the opening with a lid,
The biological treatment apparatus, wherein the fixing member can be reversed while holding and fixing the biological body.   The biological treatment apparatus according to claim 1, wherein the airtightness is airtightness under a pressure of 25 Torr. The biological treatment apparatus according to claim 1 , wherein the casing includes an exhaust port, and the exhaust port is connected to the decompression device.   The biological treatment apparatus according to claim 1, wherein the fixing member is not fixed to the casing and can be reversed independently of the casing.   The biological treatment apparatus according to claim 1, wherein the fixing member is fixed to the casing and can be reversed by inverting the casing.   The biological treatment apparatus according to any one of claims 1 to 5, wherein the casing is a cylindrical body, and the openings are opposed to both ends of the casing in the longitudinal direction.   The biological treatment apparatus according to any one of claims 1 to 6, wherein the fixing member has a cup for holding a biological body, and the cup is detachable from the fixing member.   The organism treatment apparatus according to any one of claims 1 to 7, wherein the organism is a plant.   The biological body processing apparatus of any one of Claims 1-8 used for the hydroponics of a plant body.   It is a biological treatment method performed using the biological treatment apparatus of any one of Claims 1-9, Comprising: The process which makes a biological body contact the liquid in a casing by inverting the said fixing member, Including a biological treatment method. Supplying and discharging casing, can lid sealing the opening of the casing, the casing interior located fixing member for holding and fixing the organism, the liquid prior Symbol casing having an opening for introducing and discharging the organism A gene introduction treatment device comprising supply / discharge means and a decompression device capable of reducing the pressure inside the casing ,
The gene transfer treatment apparatus, wherein the fixing member can be reversed while holding and fixing a living organism. The gene introduction treatment apparatus according to claim 11 , wherein the organism is a plant body, and the leaf stem portion of the plant is brought into contact with the liquid in the casing by inverting the fixing member. The gene introduction processing device according to claim 12 , wherein the casing has airtightness under a pressure of 25 Torr by sealing the opening with a lid. The gene introduction treatment apparatus according to any one of claims 11 to 13, which has a treatment apparatus capable of inactivating a gene contained in a liquid in a casing. Gene transfer processing device according to any one of the organism processing apparatus or Claim 11 to 14 according to any one of claims 1-9, organism growing apparatus having a growing space capable organisms Is an organism development system that is connected and arranged,
A living body breeding system comprising a transfer means that enables a living body to move between the living body growing space and the living body processing apparatus or the gene introduction processing apparatus.   The organism development system according to claim 15, wherein the organism growth apparatus has an opening, and the shape of the opening is the same as the opening of the organism treatment apparatus or the gene introduction treatment apparatus. The living body breeding system according to claim 15 or 16, further comprising an artificial light source for irradiating a living body arranged in the living body growing apparatus.

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