JPS5945873A - Vapor-phase cultivation apparatus for continuous culture of vegetable tissue - Google Patents

Abstract

PURPOSE:To culture a large amount of vegetable tissue, continuously, without damaging the tissue, by supplying a culture liquid to the vegetable tissue, and separating the excess culture liquid from the vegetable tissue dropped from the transfer means. CONSTITUTION:A culture liquid is applied continuously or intermittently to the vegetable tissue 100 on the transfer means 11, by spraying, scattering or dripping. The excess culture liquid is separated from the vegetable tissue by the separation means 13 placed under the front part of the transfer means 11. The large blocks in the dropping vegetable tissue are screened by the perforated plate 130, while small-sized blocks are transferred together with the culture liquid through the preforations and collected to the reservior 15 under the perforated plate. The small-sized blocks passed through the perforated plate 130 are recovered and recycled to the transfer means 11 as the seed. The vegetable tissue separated by the separation means 13 is discharged from the system as the final culture product by the discharging means 14.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a gas phase culture device for continuously culturing plant tissue in the entire gas phase.

(3) Conventional large-scale culture of plant cells or tissues is carried out by liquid culture similar to the fermentation method using tanks such as jar fermenters. In liquid culture, it is essential to mix and stir the culture solution using a stirring blade or air. Sliding consumes a large amount of energy, and plant cells? Destroy or damage. Such damage is particularly likely to occur in single-differentiated plant tissue systems. Therefore, mass culture of plant tissues has not yet been achieved.

An object of the present invention is to provide a vapor phase culture apparatus that can continuously culture a large amount of plant tissue without causing any physical damage to the plant tissue. Another object of the present invention is to provide a continuous gas-phase culture device in which the amount of used culture solution is minimal and the excess culture solution can be recycled and reused.

Below are drawings based on all embodiments of the present invention? This will be explained in detail with reference.

As shown in Figure 9, the apparatus of the present invention is equipped with a culture tank 1'.
Transfer means 11, culture solution supply means 12 (4), 1 separation means 13, and extraction means 14? have

The culture tank 1 further includes a storage means 15, a culture solution collection means 16, and a small tissue collection means 17.

The implantation port 10 is sealable and is provided in the ceiling of the tank 1, particularly in a corresponding portion near the rear end 112 of the transfer means 11.

The transfer means 11 is, for example, a mesh-like endless conveyor bell)
110, and this while retaining 100 plant tissues? The gas phase inside the tank 1 is transferred at a predetermined speed from one side to the other side. conveyor bell) 110
is at least one stage. Whether the method is one-stage or multi-stage can be appropriately selected depending on the growth rate of the plant tissue, the transfer rate, the space within the tank, and the like.

The culture solution supply means 12 is, for example, a nozzle 120,
The culture solution is supplied to the plant tissue 100 on the transfer means 11 either continuously or intermittently in the form of spraying, scattering, or dripping. The supply form is not limited to this. Nozzle 12
The number of zeros to be installed is not particularly limited and can be set as appropriate depending on the amount of cultured plant tissue 100, the transport distance, and the like.

The separation means 13 is disposed at the lower part of the front end of the transfer means 11 and is made of a porous hole 130 made of, for example, a stainless steel mesh. This is to separate the plant tissue and surplus culture solution that fall from the front end 111 of the transfer means 11. Among the falling plant tissues, large clumps of tissue stay on the pores &130, and small clumps of tissue together with the culture solution ′k]+! I pass through and enter the storage means 15 below. The small lump tissue that has passed through the perforated plate 130' is collected and circulated on the transfer means 11 as a new seed. This takes advantage of the fact that plant tissues are extremely stable and do not easily undergo mutations.

Since the perforated plate 130 is used to separate plant tissue according to its size, the dimensions of the mesh are important. The mesh can be selected depending on the type of plant tissue being treated and the desired size of the tissue culture as the final product. The perforated plate 130 is.

The large chunks of tissue that are separated and stay there are taken out by the next extraction means 1 due to their own weight.
It is arranged at an angle so that it can be moved to 4.

The extraction means 14 is provided adjacent to the separation means 13. The extracting means 14 is for extracting the plant tissue separated by the separating means 1.3 out of the system as a final culture. The extraction means 14 includes, for example, an inclined portion 141 provided at the extraction port 140' which can be opened and closed downward, and an air curtain mechanism 142 using sterile air to bring the inside of the system of the free exit 140 into a fully applied state. air curtain machine,
The outlet 142 completely prevents outside air from flowing into the system when the outlet 140 is fully opened.1 For example, sterile air flows into the sterile air inlet 143 through a filter by a pump (not shown), and the air inside the system is The inclined portion 1410tll' is configured to be pressurized from outside the system.

The storage means 15 is disposed below the separation means 13 and completely stores the culture solution and the small lump-like tissue 101 that are separated by the separation means 13 and flow down or fall. This storage means 15 is connected to the transfer means l from the culture solution supply means 12.
Transfer means 11 of the culture solution supplied to the plant tissue on l
It also accommodates the culture solution flowing down. For this purpose, the floor surface 102 corresponding to the transfer means 11 is appropriately inclined along the traveling direction of the transfer means 11 so that all the culture solution flowing down from the transfer means 11 can join the storage means 15. The storage means 15 is, for example, a storage tank 150, which has a culture solution outlet 152' (r) with a filter 151 on its side, and a small tissue outlet at the bottom through which the small tissue and culture solution can be lacquered. 153
have a meeting

The culture solution recovery means 16 recovers a portion of the culture solution from the mixture of the culture solution and the small lump tissue 101 stored in the storage means 15 and circulates the recovered culture solution to the culture solution supply means 12. be. This culture solution recovery means 16 connects the storage means 15 and the culture solution supply means 12. This recovery means 16 is, for example, a liquid sending g160 equipped with a culture solution circulation pump 161, one end of which is connected to the culture solution outlet 152 of the storage means 15, and the other end connected to the nozzle 120 of the culture solution supply means 12. It is connected. In order to make it possible to replenish the culture solution, the liquid supply pipe 160 can also be relayed through the culture solution tank 162.

The small lump tissue collection means 17 collects the small lump tissue 101 stored in the storage means 15 together with the culture solution from the transfer means 1.
1 near the rear end 112. This collecting means 17 is, for example, a pipe 170 provided in a small tissue circulation pump 171', and one end thereof is connected to the above-mentioned storage means 15.
The other end of the transfer means 11 is connected to the small tissue outlet 153 of the transfer means 11, and the other end thereof is opened above the rear end 112 of the transfer means 11.

Since the culture tank 1 is a tank in which all plant tissue is differentiated and grown aseptically, it has 9 heat sterilization means, temperature control means, culture solution analysis means, which are included in a normal jar fermenter.
Monitoring means etc. may be attached as necessary.

Using the device of the invention, plant tissue can be continuously cultured as follows: Sterilize the culture tank Xi with the culture medium. After cooling the culture M to a predetermined temperature, the transfer means 11. Pumps 161 and 1
71 'i drive. A desired plant tissue 1, for example a plant callus, is inoculated as whole seed tissue onto the transfer means 11 from the inlet 10 of tank 1. The inoculated tissue is transferred to the transfer means 11 while completely absorbing the supplied culture solution from the upper culture solution supply means 12.
It moves at a predetermined speed in the gas phase of tank l. The excess culture solution flows down the transfer means 11i, flows down the tank floor surface 102, and flows into the storage means 15. Plant tissue grows on the transport means 11. The grown tissue culture is transferred to the transfer means 1 along with the surplus culture solution.
from the front end 111 of 1 to the separating means 13 below. Here, the tissue culture is separated from the culture medium.

Culture solution and small lump tissue are separated by means 13? It passes through and enters the storage means 15 below. The remaining bulk tissue culture is transferred from the separation means 13 to the adjacent removal means 14 under its own weight. This large tissue culture is transferred out of the system through the outlet 140 as the final culture. At this time, the air
Since the kerf mechanism 142 is operating, there is no fear that outside air will flow into the system and contaminate the entire system.

A part of the culture solution and the small lump tissue contained in the storage means 15 is circulated to the culture solution supply means 12 by the culture solution recovery means 16 . Fresh culture solution can be replenished as needed by using the culture solution tank 162i valve.

The small lump tissue 101 in the storage means 15 is transferred to the rear end 1 of the transfer means 11 by the small lump tissue collecting means 17 together with the culture solution.
It is circulated around 12 areas. Then, as a seed tissue, it rides on the transport means 11 and moves through the gas phase in the tank.

In this way, the culture solution and plant tissues circulate within the tank 1. As for the culture solution, the culture solution tank 162
All valves were refilled with fresh culture medium as appropriate.

As for plant tissues, even if large clumps of tissue are excavated out of the system through the extraction means 14 as the final culture, small clumps of tissue 1
01 is constantly circulated onto the transport means ll as a seed tissue.

[Brief explanation of drawings]

The figure is a schematic front sectional view showing an embodiment of the device of the present invention. 1-.Culture tank, 10.. Implantation 0.11'..Transportation means. 12...Culture solution supply means, 13...Separation means, 14
... Retrieval means, 15... Storage means, 16... Culture solution collection means, 17... Small lump tissue collection means, ]00-
...Plant tissue. 101... Small lumpy tissue, 110... Porous conveyor belt. 120... Nosul, 130-... Perforated plate, 140.
...Outlet. 141... Inclined part, 142... Air curtain mechanism, 143... Sterile air inlet, 150-... Storage tank, 151-... Filter, 152... Culture solution outlet, 153... Small lump iU woven mud outlet 160...Feed-1pi pipe, 161.171...Pump. ]62...Culture solution tank, 170...Distribution-t 0
Agent: Patent Attorney Shusaku Yamamoto

Claims (1)

[Claims] 1. fll An implant port for inputting plant tissue. (2) A transfer means for transferring the plant tissue at a predetermined speed through the gas phase in the tank while holding the plant tissue. (3) a culture solution supply means that opens above the transfer means and supplies all of the culture solution to the plant tissues on the transfer means; Separation means for separating plant tissue and surplus culture solution according to the size of the plant tissue. (5) A removal means for removing the plant tissue separated by the separation means as a final culture out of the system. (6) A storage means that is disposed below the separation means and stores the mixture of the small lump tissue and the culture solution separated by the separation means and the excess culture solution flowing down during the transfer by the transfer means. (7) A culture solution recovery step of recovering a portion of the culture solution from the mixture of the culture solution and small-sized tissue stored in the storage means and circulating the recovered culture solution to the culture solution supply means. (8) A small tissue collection means for circulating the small tissue tissue stored in the storage means, together with a culture solution, over the vicinity of the rear end of the transfer means. Equipped with a culture tank with purple color. A gas phase culture device for continuously culturing plant tissues. 2. The apparatus according to claim 1, wherein the transport means is an endless conveyor belt of at least one stage. 3. The device according to claim 1, wherein the culture solution supply means is a nozzle. 4. The apparatus according to claim 1, wherein the separating means is a perforated plate. 5. The take-out means includes a sloped portion having a take-out port which can be opened and closed downward, and an air curtain mechanism using sterile air to pressurize the inside of the system at the take-out port. The plant tissue from said separation means due to its own weight. The device as set forth in claim 1, which is capable of sequentially transferring outside air out of the system through the outlet while preventing outside air from flowing into the system. 6. The storage means is a storage tank, and the tank includes:
Claim 1, further comprising a culture solution outlet with a filter on its side that is connected to the culture solution collection means, and a small tissue outlet that communicates with the small tissue collection means on the bottom thereof. Device. 7. The apparatus according to claim 11, wherein the culture solution recovery means is a liquid supply pipe that connects the storage means and the culture solution supply means via a culture solution circulation pump. 8. The small lump tissue collecting means is a pipe equipped with a small lump tissue circulation pump, one end of which is connected to the storage means and the other end opening near the rear end of the transfer means. The device according to item 1.