JP3101438B2 - Gas phase culture equipment - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a plant tissue, a plant cell,
The present invention relates to a vapor-phase culturing apparatus capable of continuously culturing a body to be cultured, such as a regenerated plant, in a gas phase, and particularly to a vapor-phase culturing apparatus capable of efficiently separating harvesting and reseeding of the body to be cultured in continuous culturing. About.

[0002]

2. Description of the Related Art Heretofore, as a method for culturing a body to be cultured such as a plant tissue, a plant cell, or a regenerated plant (hereinafter simply referred to as a body to be cultured), fermentation is performed using a tank such as a jar fermenter. It was performed by a liquid culture method according to the method. However, in the liquid culture, stirring and mixing with stirring blades, air, or the like are indispensable, and at this time, there is a possibility that the culture target may be destroyed or damaged, and such damage may occur particularly in a differentiated plant tissue system. easy. For this reason, it is very difficult to mass-cultivate the body to be cultured by liquid culture, and at present, it is not actually done.

In recent years, an apparatus capable of culturing a body to be cultivated in a gas phase in place of such a liquid culture has been proposed (for example, see Japanese Patent Publication No. 30509/1985). As shown in FIG. 3, the vapor phase culturing apparatus includes an implantation port 10 for feeding a culture object, a transfer means 11, a culture solution supply means 12,
, Separation means 13 and take-out means 14. Further, the culture tank includes a storage means 15 and a culture solution recovery means 16.
And a small-lumped culture-substance recovering means 17 and an oxygen supply means 18
And an exhaust unit 19.

[0004] According to the apparatus having the above-described structure, the transfer means 11 composed of a mesh-like endless conveyor belt 110 arranged in three stages inside the culture tank 1 causes
0 is transported from the one side to the other side in the gas phase at a predetermined speed, and the culture solution is supplied to the conveyor belt 110 by the culture solution supply means 12 comprising the nozzle 120 and the culture solution recovery means 16. The culture medium is supplied to the culture medium 100 and cultured, and the harvested large mass culture body, the small mass culture body, and the surplus culture medium are separated by the separation means 13 including the perforated plate 130 made of stainless steel net. And take-out means 14
, A continuous gas phase culture is performed in which the small-sized cultivated object collecting means 17 collects and reuses the small-sized cultivated body as a seed again. Therefore, when the culture object is vapor-phase cultured using the above-described apparatus, it is not necessary to agitate the culture object, so that physical damage to the culture object is reduced, and further, energy for stirring is unnecessary. There is an advantage of becoming.

[0005] In the above-described vapor phase culture apparatus, the culture resistance is moved on the inclined perforated plate 130, and the reseed material and the harvested material are separated according to the hole size of the perforated plate 130. The small-mass culture medium passing through the holes is collected by the small-mass culture medium collection means 17 and used for reseeding, and the large mass culture medium remaining on the perforated plate is harvested by the removal means. Therefore, in the vapor phase culture apparatus, the setting of the hole size and the inclination angle of the perforated plate 130 is important, and if the setting is slightly wrong, for example, when the hole size is small, the culture target on the perforated plate is Almost everything moves to the extraction means without going through the hole, so-called Wash out
A phenomenon occurs and continuous effective culture cannot be continued. In addition, when the pore size is large, almost all of the to-be-cultured body is recovered through the pore and used for reseeding, and cannot be harvested. Since the hole size and the inclination angle of the perforated plate differ depending on the type of the culture object, the size of the apparatus, etc., it takes a lot of time and effort to determine the appropriate hole size and the inclination angle. Therefore, there is a problem that it is difficult to perform stable continuous gas phase culture.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a vapor phase culturing apparatus which can solve the above-mentioned problems and facilitate stable continuous vapor phase culturing.

[0007]

Means for Solving the Problems The present inventors have repeatedly studied focusing on the separation method using a perforated plate in the above-described vapor phase culture apparatus, and as a result, while maintaining the perforated plate horizontally and applying vibration thereto. The present inventors have found that the above problem can be solved by adding the means, and have completed the present invention.

That is, the vapor phase culturing apparatus of the present invention is a vapor phase culturing apparatus for culturing an object to be cultured while moving the object into the gas phase. Transfer means for transferring at a speed, a culture solution supply means for supplying a culture solution to the culture medium being transferred , and separation means for separating the culture body into harvest and reseed. Means
To be moved by vibration.
Be harvested or reseed
It is characterized by having.

Hereinafter, the present invention will be described in more detail with reference to the drawings. FIG. 1 is a cross-sectional view of a gas phase culture device according to the present invention. In the figure, the difference from the above-mentioned known gas-phase culturing apparatus is that the separating means 13 is installed substantially horizontally, and the vibrating means 20 is additionally provided. The separation means 13 holds the culture object 100 falling from the terminal end of the lower belt conveyor 110 and separates the surplus culture solution from the culture medium 100. In the present embodiment, the separation plate 13 includes a perforated plate 130 made of a stainless steel net. Used. Therefore, the perforated plate 130 is connected to the transfer means 1
1, it is disposed substantially horizontally below the end of the lower belt conveyor 110. In the present invention, the separating means may be any as long as it has a function of separating the culture medium and the surplus culture solution. For example, the porous plate made of the stainless steel net, the porous side wall is formed on the porous plate It may be like a container provided with. The pore size of the perforated plate may be appropriately selected depending on the type of the culture object, the amount of culture, the size of the culture tank, and the like.

The vibrating means 2 provided in parallel with the separating means 13
Numeral 0 gives vibration in the vertical direction to the separating means 13. In this embodiment, a vibrator is used. However, the present invention is not limited to this. For example, as shown in FIG. 2, the vibrator 200 is provided at two locations on the lower surface of the perforated plate 130, and one of the vibrators (200B or 200B) is provided.
When A) is driven, the other vibrator (200A or 200B) is left stationary. The vibrating means 20 can be driven and stopped from outside the culture tank.

The driving of the vibrating means 20 is controlled by the
This is performed based on the balance between the harvest amount of 0 and the reseed amount. For example, as shown in FIG.
Is provided, the inside is observed, and the determination is made based on the amount of the culture object deposited on the perforated plate 130. Further, the apparatus of the present invention determines the drive time of the vibrating means based on the analysis of the culture time, the harvest amount, the reseed amount, etc. of the cultured body, and operates by automatic control to perform stable continuous culture. Can also.

[0012] As the culture target that can be cultured by the apparatus of the present invention, for example, at least one selected from the group consisting of plant tissues, plant cells, and regenerated plants can be used.

[0013] According to the above configuration, the culture object falling from the end of the belt conveyor is deposited on the perforated plate 130 as shown in FIG. When the vibrating means 200 is driven when a predetermined amount of the culture medium is deposited, the vibrating means 200B vibrates in the vertical direction as shown in FIG.
The culture object moves to the 200A side of the perforated plate 130 and falls. The falling culture object falls to the extraction means 14 shown in FIG.
Harvested from 4. Conversely, when the vibrating means 200A is driven, the vibrating means 200A vibrates in the vertical direction as shown in FIG.
It moves to the B side and falls. The falling culture medium falls into the storage means 15 shown in FIG. 1 and is collected as a reseed. Therefore, in the present invention, in the initial stage of culturing, the culturing is promoted by driving the vibrating means so as to collect the cultivated body as a reseed, and after a large amount of culturing is performed, the cultivated body is mainly removed from the removing means. What is necessary is just to drive a vibration means so that harvesting is possible.

[0014]

According to the above construction, the vibration means provided in the separation means applies a vertical vibration to the separation means, so that the culture object can be easily moved on the separation means. . Further, by driving the vibrating means, the culture object deposited on the separation means, when necessary,
The required amount can be dropped into the extracting means or the storing means. Therefore, a stable continuous vapor-phase culture can be easily performed while maintaining the optimal balance between the harvested amount and the reseed amount of the culture object.

[0015]

EXAMPLES Examples of the present invention will be shown below and will be described more specifically. Using the gas phase culture device shown in FIG.
Eleuthero belonging to (Araliaceae) (scientific name: Acanthopan
ax senticosus) callus derived from leaves
Continuous cultivation was performed at 0. In this example, 1 ppm of IBA was used as auxin in the MS medium in which the nitrogen source was modified.
m, 0.1 ppm of kinetin was added as a cytokinin to prepare a culture solution. In addition, as a sugar source, sucrose 3
%It was used. 2 liters of the above culture solution was used for the gas phase culture apparatus
And sterilized in the culture tank 1. In addition to this callus 100
Conveyor belt 1 from implant 10 by 150 g
10 were inoculated uniformly. After closing the implantation port 10, the conveyor belt 110 and the vibrator 200 </ b> A were driven, and the culture solution was sprayed from the nozzle 120. The required amount of the culture solution was absorbed by the culture subject 100, and the remainder dropped into the storage tank 150 as droplets. In this way, after keeping the inside of the tank at 25 ° C. and performing gas phase culture of the callus 100 for 2 weeks, the drive of the vibrator is switched to 200B, and about 5/6 of the total callus is taken out to the extraction means 14 side. Dropped and harvested. Next, the drive of the vibrator is switched to 200A, and the remaining callus 100 is dropped to the storage tank side and collected as a reseed. It could be done continuously. The callus induction used in this embodiment is disclosed in detail in the above-mentioned Japanese Patent Application Laid-Open No. 1-228464.

[0016]

According to the gas phase culture apparatus of the present invention, the following effects can be obtained. According to the above configuration, since the vibration in the vertical direction is applied to the separating means by the vibrating means provided in parallel with the separating means, the inoculated body can be easily moved on the separating means. In addition, by driving the vibrating means, it becomes possible to drop a required amount of the culture to be deposited on the separating means to the extracting means or the storing means when necessary. Therefore, a stable continuous vapor-phase culture can be easily performed while maintaining the optimal balance between the harvested amount and the reseed amount of the culture object.

[Brief description of the drawings]

FIG. 1 is a sectional view showing an example of a gas phase culture device according to the present invention.

FIG. 2 is a diagram illustrating an operation of a separating unit by a vibrating unit.

FIG. 3 is a cross-sectional view showing a conventional gas phase culture device.

[Explanation of symbols]

 Reference Signs List A gas phase culture apparatus 1 culture tank 11 transfer means 12 culture solution supply means 13 separation means 20 vibration means 100 culture object

Continuation of the front page (56) References JP-A-3-85451 (JP, A) JP-B-60-30509 (JP, B2) (58) Fields investigated (Int. Cl. ⁷ , DB name) C12M 1 / 00 C12M 3/00

Claims (2)

(57) [Claims] 1. A vapor phase culturing apparatus for culturing a body to be cultured while moving the body into a gas phase, comprising: a transfer means for transferring the body to be cultured into a gas phase in a culture tank at a predetermined speed; A culture solution supply means for supplying a culture solution to the medium to be cultured therein, and separation means for separating the culture body into harvest and reseed.
Vibration means are installed in the step, and the culture
Move the body by vibration and separate it into harvest or reseed
A vapor phase culturing apparatus characterized in that it is configured to be capable of causing the culturing. 2. The method according to claim 1, wherein the transferring means is configured to transfer the object to be cultured in a culture tank.
A belt conveyor for transferring at a predetermined speed in the gas phase
Wherein the separating means comprises a belt conveyor.
2. A perforated plate which is disposed substantially horizontally below the terminal end.
The gas-phase cultivation apparatus according to the above.