JPH06165624A - Cultivation of plant body and bioreactor - Google Patents

Abstract

PURPOSE:To improve the absorption efficiency of nutrient to a plant body, increase the dissolved oxygen concentration, promote the growth of a plant body and enable the cultivation of a plant without using complicate operations such as transplantation of the plant body. CONSTITUTION:A plant body S is put into a culture vessel 10 of a bioreactor. The culture liquid W in a tank 15 is sucked by a pump 23, supplied through an ejection pipe 25 to the top of the culture vessel 10 and stored in the vessel 10. When the liquid level reaches the top of a siphon pipe 20, the culture liquid in the culture vessel 10 is flowed down through the siphon pipe 20 into the tank 15 at a stroke by the pressure difference between the culture vessel 10 and the tank 15. The plant body is subjected to the repetition of the immersion in the culture liquid and the exposure to the atmosphere and the growth of the plant body is promoted by the stimulation caused by the repeated immersion and exposure. Furthermore, the dissolved oxygen concentration in the culture liquid is maintained to a sufficiently high level by the relative transfer of the culture liquid and, accordingly, the plant body can be activated, the absorption efficiency of the nutrient to the plant body is improved and the growth of the plant body is promoted.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for cultivating a plant such as a plant and a bioreactor for culturing the plant.

[0002]

2. Description of the Related Art Conventionally, a method of culturing a plant will be described, for example, in the case of culturing a plant in which a part of the plant is cut off. As shown in FIG. A solid medium 2 such as agar is formed, the plant S is placed on the medium 2, the lid 3 is placed on the medium S, and the plant S is left under appropriate temperature and humidity conditions. As a result, the plant S proliferates to form a so-called callus. Next, this callus is divided, and as shown in FIG. 5B, it is replanted in another container 1 provided with a medium 4 in which a plant hormone for germination is mixed. If germinated in germination medium 4, then FIG.
As shown in (c), it is transplanted to another container 1 provided with a medium 5 in which a plant hormone for rooting is mixed. Then, the rooting medium 5 is allowed to root and grow.

[0003]

However, the above-mentioned conventional method for culturing a plant has a problem that the growth of the plant is slow. The reason for this is that the tissue-cultured plant body has a relatively insufficient ability to absorb nutrients, and thus the absorption of nutrients is slowed by contact with a solid phase such as agar. As the nutrients around the plant are consumed because the medium is in the solid phase, the supply of new nutrients becomes insufficient,
It causes nutrient deficiency. Waste products generated from the roots of the plant body are accumulated in the medium, and the waste products inhibit the growth of the plant body. Dissolved oxygen in the medium is limited, and sufficient oxygen cannot be supplied to the plant body. Since the type of medium must be changed according to the growth process of the plant, the plant must be replanted each time, but the root of the plant is damaged during the replanting, and this damage Inhibits the growth of plants. It is because of such reasons.

Further, in the above-described conventional method for culturing a plant, the type of medium has to be changed according to the developmental stage of the plant, so that the plant must be replanted each time. Therefore, there is also a problem that the labor for growing the plant is complicated accordingly.

Therefore, an object of the present invention is to improve the absorption efficiency of nutrients, increase dissolved oxygen, and accelerate the growth of plants. Moreover, the complicated work such as replanting of plants is required. It is intended to provide a method for culturing a plant and a bioreactor that can be cultivated without performing the above.

[0006]

[Means for Solving the Problems] A method for culturing a plant according to the present invention for solving the above-mentioned problems is a method for culturing a plant in a culturing method in which the plant is placed in a culture container and cultivated. The solution is placed in a culture vessel to immerse the plant in the culture solution, the culture solution is discharged from the culture vessel to expose the plant to the atmosphere, and the immersion and exposure are repeated to culture the plant. is there.

Further, the bioreactor of the present invention comprises a sealed culture container in which a plant is placed, a sealed tank disposed below the culture container and in which a culture solution for culturing the plant is placed, The siphon tube, which has one end opened into the tank and the other end opened into the culture container and the path leading to the open other end in the culture container extends upward in the culture container and then bends downward, and the culture solution in the tank It is provided with a pump for sucking and discharging, a discharge pipe for supplying the culture solution discharged from the pump into the culture container, and a communication pipe for connecting the upper part of the culture container and the upper part of the tank.

It is effective to provide a stirring device for stirring the culture solution in the tank to dissolve the air in the tank in the culture solution.

Further, it is effective that the upper part of the culture vessel is provided with an air-permeable porous body.

[0010]

According to the plant culture method of the present invention thus constructed, the plant is repeatedly immersed in the culture solution and exposed from the culture solution in the culture container. In addition, during the immersion and exposure of the plant body, the relative movement of the culture solution makes it easier for air to be mixed into the culture solution, thereby increasing the dissolved oxygen. Further, the culture solution can be exchanged or modified according to the growth process of the plant body, and since the culture solution is a liquid, the exchange or modification can be easily performed.

According to the bioreactor having the above structure, when the plant is placed in the culture container and the pump is driven in this state, the culture solution in the tank is sucked and the upper part of the culture container is discharged through the discharge pipe. It is supplied from and accumulates in the culture container. Then, when the liquid level reaches the upper end of the siphon tube, a difference occurs between the internal pressure of the culture container and the internal pressure of the tank, so that the culture solution in the culture container flows down the siphon tube all at once. Then, since the culture solution is supplied from the upper part of the culture container, the culture solution is accumulated again in the culture container. In this process, the plant body is immersed in the culture solution when the culture solution is accumulated in the culture container, and is exposed to the inside air of the culture container when the culture solution is discharged from the culture container. Then, this immersion and exposure are repeated. Further, since the culture solution circulates, air is easily mixed in the culture solution.

When the agitating device for agitating the culture solution in the tank is provided, the culture solution is agitated. Therefore, in addition to circulation of the culture solution, air entrainment is promoted and dissolved in the culture solution. The amount of oxygen is increased.

When the culture container is provided with an air-permeable porous body, fresh air is supplied into the culture container and the temperature inside the culture container is adjusted.

[0014]

The plant culturing method and bioreactor according to the embodiments of the present invention will be described below with reference to the accompanying drawings.
The plant culture method is realized by the bioreactor according to the example. FIG. 1 shows a bioreactor according to the example.

In FIG. 1, 10 is a sealed culture container in which the plant S is placed. This culture vessel 10 is a glass tube which is vertical and whose upper and lower openings are sealed with stoppers. The upper stopper 11 of the upper part of the culture container 10 is a porous body having air permeability and pores that prevent invasion of various bacteria, for example,
It is composed of a sponge-like silicon stopper. Culture vessel 1
The lower plug 12 of the lower part 0 is formed of a non-air-permeable rubber plug.

Reference numeral 15 denotes a sealed tank which is disposed below the culture container 10 and in which a culture solution W for culturing the plant S is put. The tank 15 is formed by sealing an opening of a glass container-shaped tank body with a non-air-permeable rubber stopper 16. First and second insertion holes 17 and 18 are formed on the side portion of the body of the tank 15 so as to communicate with each other and sealed with a stopper.

Reference numeral 20 denotes a siphon tube penetrating the rubber stopper 16 of the tank 15 and the lower stopper 12 of the culture container 10. One end 21 is open to the upper part of the tank 15 and the other end 22 is the culture container 1.
The path extending to the inner lower portion of the culture container 10 and extending to the open other end 22 in the culture container 10 extends upward in the culture container 10 and then bends and extends downward. The other end 22 and the bottom (bottom stopper 1
The distance L (FIG. 2) from the upper surface of 2 is set appropriately.

Reference numeral 23 is a pump for sucking and discharging the culture solution W in the tank 15. 24 is a suction pipe connected to the pump 23, which penetrates the rubber stopper 16 of the tank 15,
The suction port reaches the lower part of the tank 15. Reference numeral 25 denotes a discharge pipe connected to the pump 23, which penetrates the upper stopper 11 of the culture container 10, the discharge port 26 is located in the upper part of the culture container 10, and the culture solution W discharged from the pump 23 is supplied to the culture container. 1
It is supplied from above from 0. Reference numeral 27 denotes a drive control unit of the pump 23, which adjusts the suction / discharge amount of the culture solution W and is programmed to appropriately adjust the drive stop time of the pump 23.

Reference numeral 28 is a communication pipe that connects the upper part of the culture container 10 and the upper part of the tank 15 to each other.
6 and the upper stopper 11 of the culture container 10 are penetrated.

Reference numeral 30 is a stirring device for stirring the culture solution W in the tank 15 so that the air in the tank 15 is dissolved in the culture solution W. The stirrer 30 includes a stirrer 31 provided in the tank 15 and a stirrer base 32 on which the tank 15 is mounted and which magnetically rotates the stirrer 31.

Reference numeral 33 denotes a supply device for supplying the culture solution W into the tank 15, and the culture solution W in the flask 34 is pumped by the pump 3
5, and the gas is discharged into the tank 15 through the discharge pipe 35a penetrating the plug of the first insertion hole 17 of the tank 15. Reference numeral 36 denotes a discharge device 36 for discharging the culture solution W in the tank 15, and a pump 37 for pumping the culture solution W in the tank 15 through a suction pipe 36a penetrating the stopper of the first insertion hole 17 of the tank 15. And suck into the flask 38.
Each of the flasks 34, 38 is provided with a flow pipe 39 for circulating the air inside the flasks 34, 38. 39a is an air filter interposed in the flow pipe 39.

In the second insertion hole 18 of the tank 15, an electric conductivity meter (EC meter) 40 for measuring the nutrient concentration of the culture solution W in the tank 15 and a dissolved oxygen for measuring the amount of dissolved oxygen. An oxygen measuring device (DO meter) 41, a PH meter 42, etc. are attached. Reference numeral 43 denotes a display unit 43 that displays the measured value of each instrument.

Next, the case of cultivating the plant S using this bioreactor will be described. As the plant body S, for example, a part of “asparagus” that has been cut off is cultured. As shown in FIG. 2, the plant S together with its holder 44 is put in the culture container 10 in advance. As the holding body 44, for example, a large number of jelly-like and beads-like ones formed by dropping sodium alginate into calcium chloride are used. This holder 44 has a specific gravity larger than that of the culture medium W, a normal particle size of 1 mm to 4 mm, and a siphon tube 2
Greater than 0 inner diameter. In addition, in the tank 15 in advance,
For example, a culture medium W mixed with a plant hormone for callus production (for example, α-naphthalene acetic acid as auxin)
Add an appropriate amount. Further, the flask 3 of the supply device 33
A similar culture solution W is also placed in 4.

In this state, the drive controller 27 drives the pump 23 and the stirring device 30. As a result, the culture solution W in the tank 15 is sucked through the suction pipe 24, supplied from the upper portion of the culture container 10 through the discharge pipe 25, and accumulates in the culture container 10 from the bottom surface thereof. Then, as shown in FIG. 3, when the liquid level reaches the upper end of the siphon tube 20, a difference occurs in the internal pressure of the culture container 10 and the internal pressure of the tank 15, and the culture container 10 and the tank 15 are connected by the communication pipe 28. Since they are communicated with each other, the culture solution W in the culture container 10 travels down the siphon tube 20 and flows down into the tank 15 all at once. Then, since the culture solution W is supplied again from the upper part of the culture container 10, as shown in FIG.
The culture solution W accumulates from the 0 bottom surface. In this process, the plant body S is immersed in the culture solution W when the culture solution W is accumulated in the culture container 10, and is exposed to the inside air of the culture container 10 when the culture solution W is discharged from the culture container 10. . Then, this immersion and exposure are repeated.

In this process, since the agitating device 30 is driven, the culture solution W is agitated and also agitated when flowing down from the siphon tube 20. As a result, the air in the tank 15 and the culture container 10 is mixed with the culture solution W, so that the dissolved oxygen amount of the culture solution W is maintained in a substantially saturated state. Further, on the upper part of the culture container 10, a breathable upper stopper 1
1, the supply of fresh air is ensured. Therefore, it is possible to prevent the situation in which the temperature and humidity are adjusted and the temperature and humidity are high and the plant S is adversely affected. In addition, this bioreactor is a closed system, and since the upper plug 11 of the upper part of the culture container 10 is breathable but has very fine holes, it prevents the invasion of various bacteria from the outside, and the environment of the plant S is good. Kept in.

In this process, the plant S is in the culture solution W.
It absorbs nutrients and hormones inside and forms callus. In this case, since the plant body S is in contact with the liquid called the culture medium W, it is in good contact with the nutrients, and since the dissolved oxygen amount is substantially saturated, the oxygen amount is sufficiently secured. , The nutrients are absorbed more efficiently, and the faster the growth. In particular, the plant body S is repeatedly soaked and exposed to the culture solution W periodically, which serves as a stimulus to promote the growth. Further, since the culture solution W is circulated, the nutrients around the plant S are sufficiently supplied, and the situation in which the nutrients are insufficient can be prevented.

Further, in this process, the nutrient concentration, the dissolved oxygen amount and the PH of the culture solution W in the tank 15 are measured by the respective instruments, and the measured values are displayed on the display section 43. for that reason,
Changes in the culture solution W such as the consumption state of nutrients can be seen. Then, if it is determined from the measured value displayed on the display unit 43 that the nutrients in the culture medium W have been consumed, the pump 37 of the discharge device 36 is driven to discharge the culture medium W to some extent and supply it. The pump 35 of the device 33 may be driven to supply a new culture solution W. As a result, the supply of nutrients and hormones becomes sufficient, and in this respect as well, growth can be accelerated. In this case, the tank 15 and the culture container 1
While keeping 0 closed, nutrients and hormones can be supplied and the culture solution W can be kept in a constant state, so that the plant S is prevented from being damaged. It should be noted that only the nutrients and hormones may be supplied from the first insertion hole 17 without discharging and supplying the culture solution W.

Next, when callus is formed, the pump 23 is stopped and the culture solution W containing seedling-forming hormone is replaced. In this case, first, the pump 37 of the discharge device 36
Is driven to discharge the culture solution W, and the culture solution W containing a plant hormone (eg, kinetin as cytokinin) is placed in the flask 34 of the supply device 33, and the pump 35 is driven to supply a new culture solution W. good. Further, the plant hormone may be added from the first insertion hole 17 to the culture solution W used so far without discharging the culture solution W.
In this case, with the tank 15 and the culture container 10 closed,
Since the culture solution W can be exchanged or modified, damage to the plant S can be prevented. Further, in this case, the culture solution W can be exchanged or modified without replanting the plant S, so that the work is easily performed.

Then, the pump 23 is driven again. As a result, the plant S is immersed in another culture solution W and exposed to the inside air of the culture container 10, and the immersion and exposure are repeated. In this process, the plant S absorbs nutrients and plant hormones in the culture solution W to form seedlings. Also in this case, since the plant S is in contact with the liquid called the culture medium W, it is in good contact with the nutrients, and moreover, the dissolved oxygen content is almost saturated, so that the oxygen content is sufficiently secured. Therefore, the absorption efficiency of nutrients is improved, and the faster the growth takes place. If nutrients, plant hormones, etc. in the culture solution W are consumed, the pump 37 of the discharge device 36 is driven to discharge the culture solution W to some extent and the pump 35 of the supply device 33 is driven in the same manner as above. It may be driven to supply a new culture solution W.

Next, if seedlings are formed, the pump 23
Is stopped and the culture medium W for rooting, for example, one having a low auxin concentration is exchanged. In this case, first, the discharge device 3
6, the culture medium W is discharged by driving the pump 37, the culture medium W for rooting is put in the flask 34 of the supply device 33, and the pump 35 is driven to supply a new culture medium W. Even in this case, since the culture solution W can be exchanged or modified while the tank 15 and the culture container 10 are closed, the plant S is prevented from being damaged. Further, since the culture solution W can be replaced or modified without replanting the plant body S, the work is easily performed.

Then, the pump 23 is driven again. As a result, the plant S is immersed in the rooting culture solution W and exposed to the inside air of the culture container 10, and this immersion and exposure are repeated. In this process, as shown in FIG. 4, the plant body S absorbs the nutrients in the culture solution W and roots and grows. In this case, the root extends between the holding bodies 44, so that the holding is ensured and the above-ground portion can be stably grown upward. Further, since the plant body S is in contact with the liquid called the culture solution W, the contact with the nutrients is good,
Moreover, since the amount of dissolved oxygen is almost saturated, the amount of oxygen is sufficiently secured, so that the absorption efficiency of nutrients is improved and the growth is accelerated accordingly. In particular, when the plant body S is periodically immersed in the culture solution W, it becomes wet and dry repeatedly, which stimulates the growth of the vascular bundle (conduit, phloem) of the plant. . Therefore, the nutrient absorption capacity of the root is fully developed, and the nutrient absorption is accelerated. Further, since the culture solution W is circulated, the nutrients around the plant S are sufficiently supplied, and the situation in which the nutrients are insufficient can be prevented.

At this stage, the amount of waste products and growth-inhibiting substances emitted from the plant S, especially roots, increases, but since the culture solution W circulates, these waste products and growth-inhibiting substances diffuse. Therefore, the situation of inhibiting the growth of roots is prevented, and the growth is accelerated also in this respect. If the nutrients in the culture medium W are consumed,
Similar to the above, a new culture solution W may be supplied using the discharge device 36 and the supply device 33.

In the above embodiment, the holder 44 for the plant S was placed in the culture container 10, but the culture can be performed without necessarily holding the holder 44. Further, the shape and size of the culture container 10 and the tank 15 are not limited to those described above, and may be changed as appropriate. Furthermore, in the above-mentioned embodiment, "asparagus" is used as the plant S, but the plant S is not limited to this and may be applied to all kinds of plant S. The components of the culture solution W are not limited to those described above and may be changed as appropriate.

[0034]

As described above, according to the plant culturing method and the bioreactor of the present invention, the plant is cultivated by repeatedly immersing the plant in the culture solution and exposing the plant to the atmosphere. Therefore, since the plant comes into contact with a liquid called a culture solution, the contact with the nutrients is improved, and since the plant is repeatedly soaked and exposed to the culture solution, this stimulates the growth. It In addition, the relative movement of the culture broth facilitates the incorporation of air into the culture broth, ensuring a sufficient amount of dissolved oxygen in the culture broth and activating the plant body. Therefore, the absorption efficiency of the nutrients of the plant can be improved, and the growth can be accelerated.

Further, since the culture medium moves relatively, nutrients around the plant can be sufficiently supplied to prevent a situation where the nutrient is insufficient, and at the same time, waste products and growth produced from the plant can be prevented. Since the inhibitory substance diffuses, it is possible to prevent the situation in which the growth of the plant is inhibited, and also in this respect, the growth can be accelerated.

Further, the culture medium can be exchanged or modified according to the growth process of the plant, and since the culture medium is a liquid, the exchange or modification can be easily carried out,
Therefore, unlike the conventional case, it is not necessary to replant a plant body, and accordingly, there is an effect that complexity is reduced and workability and management can be facilitated.

Further, according to the bioreactor of the present invention,
Since it is a closed system using the siphon principle, invasion of various bacteria can be prevented and the plant can be protected from the influence of various bacteria.

Furthermore, in the case where a stirring device for stirring the culture solution in the tank is provided, the culture solution can be stirred to promote the entrainment of air. Therefore, in addition to the entrainment of air by circulating the culture solution, The amount of dissolved oxygen in the culture solution can be increased.

Further, when an air-permeable porous body is provided on the upper part of the culture container, fresh air is slowly supplied into the culture container, and water vapor can also flow in and out.
It is possible to prevent the situation where the temperature and humidity inside the culture vessel are adjusted and the temperature and humidity are high and the plants are adversely affected.

[Brief description of drawings]

FIG. 1 is a perspective view showing a bioreactor according to an embodiment of the present invention.

FIG. 2 is a cross-sectional view showing a bioreactor according to an embodiment of the present invention together with the supply state of the culture solution.

FIG. 3 is a cross-sectional view showing a bioreactor according to an example of the present invention together with another supply state of the culture solution.

FIG. 4 is a cross-sectional view showing a bioreactor according to an example of the present invention together with a growing state of a plant.

FIG. 5 is a diagram showing a conventional method for culturing a plant.

[Explanation of symbols]

 S Plant W Culture medium 10 Culture vessel 11 Upper stopper 12 Lower stopper 15 Tank 16 Rubber stopper 17 First insertion hole 18 Second insertion hole 20 Siphon pipe 21 One end 22 Other end 23 Pump 24 Suction pipe 25 Discharge pipe 26 Discharge Outlet 27 Drive control unit 28 Communication pipe 30 Stirring device 33 Supply device 36 Discharge device 40 Electric conductivity meter (EC meter) 41 Dissolved oxygen measuring device (DO meter) 42 PH meter 43 Display unit

Claims (4)

[Claims] 1. In a culture method for culturing a plant in a culture vessel, a culture solution for culturing the plant is placed in the culture vessel, the plant is immersed in the culture solution, and the culture solution is placed in the culture vessel. A method for cultivating a plant, which comprises culturing the plant by discharging it from the plant to expose the plant to the inside air, and repeating the immersion and the exposure. 2. A sealed culture container in which a plant is placed, a sealed tank which is disposed below the culture container and holds a culture solution for culturing the plant, and one end of which is opened into the tank. A siphon tube with the other end opening into the culture container and a path leading to the open other end in the culture container extending upward in the culture container and then bending and extending downward, a pump for sucking and discharging the culture solution in the tank, and a pump A bioreactor comprising: a discharge pipe for supplying the culture solution discharged from the inside of the culture container, and a communication pipe for connecting the upper part of the culture container and the upper part of the tank. 3. The bioreactor according to claim 2, further comprising a stirrer for stirring the culture solution in the tank to dissolve the air in the tank in the culture solution. 4. The bioreactor according to claim 2, further comprising an air-permeable porous body on the upper part of the culture vessel.