CN110896852A - Potential energy driven intermittent immersion type bioreactor - Google Patents
Potential energy driven intermittent immersion type bioreactor Download PDFInfo
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- CN110896852A CN110896852A CN201811545726.8A CN201811545726A CN110896852A CN 110896852 A CN110896852 A CN 110896852A CN 201811545726 A CN201811545726 A CN 201811545726A CN 110896852 A CN110896852 A CN 110896852A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention discloses a potential energy driven intermittent immersion bioreactor, which comprises a tissue culture assembly, a liquid assembly, a first lifting assembly and a second lifting assembly, wherein the tissue culture assembly is communicated with the liquid assembly through a connecting pipeline and is arranged at the upper part of the first lifting assembly; the liquid assembly is arranged on the upper part of the second lifting assembly, and culture solution in the liquid assembly is injected into the tissue culture assembly or is discharged into the liquid assembly through the lifting of the first lifting assembly and the second lifting assembly, so that the plant tissue in the tissue culture assembly is subjected to intermittent immersion treatment. The principle of the bioreactor of the present invention is to directionally drive the liquid culture medium by gravity after changing the height (potential energy) of the vessel, thus distinguishing it from other types of batch submerged bioreactors. The bioreactor has the advantages of simultaneously operating a plurality of groups of bioreactors, having relatively simple structure, easy operation, convenient unified management and reduced misoperation. Is especially suitable for large-scale tissue culture production.
Description
Technical Field
The invention relates to a bioreactor for plant tissue culture, in particular to an intermittent immersion type bioreactor for plant tissue culture, belonging to the technical field of plant material tissue culture and plant development.
Background
Batch Immersion Bioreactors (TIBs) are a system developed for plant tissue culture and plant secondary metabolite research in recent years by international research.
The plant tissue culture and rapid propagation technology is superior to the traditional seed propagation and seed stem propagation because of the characteristics of rapid propagation of the seedling through the explant, maintenance of the genetic characteristic of the parent, acquisition of the seedling without virus and the like, thereby achieving the rapid development and full utilization of the technology. The research on large-scale liquid immersion tissue culture is numerous at home and abroad, and the intermittent immersion bioreactors for plant tissue culture and rapid propagation mainly have four types: an APCS intermittent complete immersion culture system, a peristaltic pump driven intermittent semi-immersion culture system, an air pump driven intermittent immersion bioreactor and TIBs intermittent immersion bioreactor.
The APCS intermittent complete immersion culture system consists of 4 parts: the whole system realizes the regular supplement and drainage of liquid under the drive of the impeller, and saves the successive transfer operation of frequently updating the culture solution; the peristaltic pump drives the intermittent semi-submerged culture system to be an APCS system improvement, a timer is added, the peristaltic pump and a time control switch are utilized to achieve automatic intermittent updating of nutrient solution, and intermittent semi-submerged culture is conducted on tissue culture seedlings; above two kinds of culture systems all utilize external mechanical power as the direct drive power of liquid medium, utilize the control procedure to realize the intermittent type of nutrient solution and supply and drain, have reduced labour's consumption, have reduced manufacturing cost, but because it is long consuming time to change the nutrient solution, the operation is complicated, pollutes seriously and space restriction problem for it receives the restriction of certain degree on popularization and application. The air pump drives the intermittent immersed bioreactor to have two containers to make up, there are nutrient solution storage containers below, above there are tissue culture seedling culture containers, there are pipelines to link up in the middle, utilize the atmospheric pressure that the air pressure pump produces as the direct driving force of the liquid culture medium, oppress the liquid culture medium to flow into the tissue culture seedling culture container from the nutrient solution storage container through the central pipeline, cultivate, there is flow of the gas at the same time, realize the renewal of the air in the culture container, when the gas enters the nutrient solution reservoir, in order to prevent the polluted gas from needing to be filtered through the polytetrafluoroethylene filter membrane of 0.22 μm, but the system is small, the structure is complicated, the price is high, these conditions have limited and promoted and applied; the TIBs intermittent immersion bioreactor is a set of equipment integrating plant tissue culture and plant secondary metabolite research, and utilizes a liquid culture medium to perform intermittent culture on plant tissue culture seedlings by using filtered air pressure as a direct driving force of the liquid culture medium, and the main components are as follows: a tissue culture seedling culture container, a nutrient solution storage container, an air compressor, a needle filter with a filter membrane aperture of 0.22 mu m, a time control switch and an electromagnetic valve. The two containers are connected through a silicon tube, gas is sterilized through a needle filter with the aperture of a filter membrane of 0.22 mu m, the supply and the drainage of nutrient solution are realized by utilizing the gas pressure of an air compressor, and the control of the intermittent frequency is realized by controlling an electromagnetic valve through a time switch to realize the time and the direction of air flow. The system avoids the damage of vitrification deformity and secondary metabolites caused by a long period in the liquid culture process, effectively prevents the deposition of nutrient substances and the accumulation of harmful substances, enables the tissue culture seedling to more effectively utilize the nutrient components of a culture medium, and provides a good environment for plants in the liquid culture process.
The existing intermittent immersion type bioreactor has the defects of complex operation, serious pollution and space limitation (such as an APCS intermittent complete immersion culture system and a peristaltic pump driven intermittent semi-immersion culture system) or small volume, complex structure and high price (such as an integral air pump driven intermittent immersion type bioreactor). The air pump drives the intermittent immersion type bioreactors, each group of bioreactors needs an independent air compression pump (uneven air pressure is easily caused when one pump is used for multiple purposes), and sterile air is used for directly applying pressure to the liquid culture medium. These can result in increased bioreactor cost and contamination of plant material (e.g., integrated and split air pump driven batch immersion bioreactors). Bioreactors that are costly and rely on independent operating power are also not conducive to scale-up to uniform standards.
The above-mentioned 4 types of bioreactors are driven by external force (mechanical force, air pressure) directly acting on the liquid culture medium, and thus have the disadvantages of complicated structure, high manufacturing cost, difficult operation, easy contamination, etc. The bioreactor is fundamentally different in that after the potential energy (position height) of the container is changed by external force, the liquid culture medium is directionally driven by the gravity, and the bioreactor has the advantages of simple structure and easiness in operation, each power system can simultaneously operate a plurality of groups of bioreactors, so that unified management is facilitated, and the opportunities of misoperation and plant material pollution are reduced. Is particularly suitable for standard large-scale production.
Disclosure of Invention
The invention aims to provide a potential energy driven intermittent immersion bioreactor aiming at the technical defects of bioreactors used in the existing plant tissue culture process. The tissue culture assembly and the liquid assembly are respectively arranged on the two groups of lifting platforms (plates). The flow of the liquid culture medium between the tissue culture assembly and the liquid assembly is controlled by the change of the relative height between the tissue culture assembly and the liquid assembly. The bioreactor of the invention changes the potential energy (position height) of the container by external force, and directionally drives the liquid culture medium by means of the gravity, in the process of tissue culture, the liquid culture medium flows between the tissue culture assemblies and the liquid assemblies through the change of the relative height between each group of tissue culture assemblies and the liquid assemblies, thereby realizing the intermittent immersion of plant materials placed in the tissue culture assemblies. The reaction device has simple structure and easy operation, each power system can simultaneously operate a plurality of groups of bioreactors (different sizes), thereby being convenient for overall management and being suitable for standardized production.
To achieve the object of the present invention, the present invention provides a potential energy driven intermittent submerged bioreactor comprising: group banks up subassembly, liquid subassembly and first lifting unit, second lifting unit with earth, wherein:
the tissue culture assembly is used for placing plant materials to be subjected to tissue culture treatment, and the plant materials are subjected to tissue culture in the tissue culture assembly; the liquid component is used for storing liquid culture medium; the tissue culture assembly is communicated with the liquid assembly through a connecting pipeline;
the first lifting assembly is used for lifting or lowering the tissue culture assembly; the second lifting assembly is used for lifting or lowering the liquid assembly, and along with the lowering of the first lifting assembly or the lifting of the second lifting assembly, the culture solution in the liquid assembly is injected into the tissue culture assembly; along with the rising of first lifting unit or the reduction of second lifting unit for the culture solution in the group banks up the subassembly with earth and flows into the liquid subassembly, realizes that the culture solution is to the intermittent type submergence of the material of treating in the group banks up the subassembly with earth and handle.
The tissue culture assembly is communicated with the liquid assembly through a connecting pipeline, and culture solution in the liquid assembly is injected into the tissue culture assembly through the connecting pipeline by the lowering of the first lifting assembly or the rising of the second lifting assembly; through the rising of first elevating system or the reduction of second elevating system, the culture solution of group in the subassembly of cultivateing discharges into in the liquid subassembly through connecting tube, carries out intermittent type submergence processing to the plant tissue of group in the subassembly of cultivateing.
Particularly, the first lifting assembly keeps the height unchanged, and the culture solution in the liquid assembly is injected into the tissue culture assembly through the lifting of the second lifting assembly; through the reduction of second lifting unit for the culture solution in the group banks up the subassembly with earth flows into liquid subassembly, realizes that the culture solution is to the intermittent type submergence of the material of treating in the group banks up the subassembly with earth and handles. Or the second lifting component keeps the height unchanged, and the culture solution in the liquid component is injected into the tissue culture component through the reduction of the first lifting component; through the rising of second lifting unit for the culture solution in the group banks up the subassembly with earth flows into liquid subassembly, realizes that the culture solution is to the intermittent type submergence of waiting to handle the material in the group banks up the subassembly with earth.
Wherein, group banks up subassembly including incubator and incubator support with earth, wherein the incubator is placed and is fixed the upper portion of incubator support with earth.
In particular, the incubator is used for placing plant tissue culture materials, such as plant callus, plant embryogenic tissue or mature somatic embryos and the like; the incubator support plate is flat, the incubator is placed on the upper portion of the incubator support plate, the incubator support is horizontally placed on the upper portion of the first lifting component, and the incubator support and the incubator placed on the upper portion of the incubator support are driven to ascend or descend along with ascending or descending of the first lifting component.
The top of the incubator is provided with an incubator cover used for sealing the incubator body, the incubator cover is provided with an air filter, and gas entering and exiting the incubator is filtered to prevent pollution.
Particularly, the middle lower part of the incubator is provided with an incubator liquid inlet/outlet hole, and the size of the liquid inlet/outlet hole is matched with that of the connecting pipeline.
Particularly, the liquid inlet/outlet hole is formed at a position close to the bottom of the incubator, so that the complete discharge of the culture solution in the incubator is facilitated.
In particular, the incubator further comprises an internal support box disposed within the chamber for receiving plant material to be treated.
The size of the horizontal section of the inner support box is matched with that of the horizontal section of the incubator, and the top of the inner support box is clamped and fixed with the top of the incubator and is level with the top of the incubator in the same horizontal plane.
Particularly, the ratio of the height of the inner support box to the height of the incubator is 1/2-2/3.
Particularly, the bottom of the internal support box is provided with a plurality of liquid flowing holes for culture solution to flow freely and to enter and exit the internal support box freely.
In particular, the plant tissue to be treated is placed on the bottom surface of the inner support box of the incubator or on a support layer laid on the bottom surface of the inner support box.
In particular, the top of the incubator is clamped and fixed with the top of the incubator, and is level with the top of the incubator and in the same horizontal plane.
In particular, the incubator further comprises a support plate arranged within the chamber for placing plant material to be treated.
Wherein, the shape and the size of the support plate are matched with the shape and the size of the horizontal section of the incubator.
In particular, the support plate is arranged close to the bottom of the incubator at a distance from the bottom of the incubator.
Particularly, the ratio of the height of the support plate from the bottom of the incubator to the height of the incubator is 1/4 to 1/2.
The plant material of the present invention may be placed directly on the bottom of the incubator.
Wherein, the liquid subassembly includes liquid reserve tank and liquid reserve tank support, wherein the liquid reserve tank is placed and is fixed on the upper portion of liquid reserve tank support.
In particular, the culture solution tank is used for storing culture solution for plant tissue culture, such as various liquid culture media and the like; the liquid reserve tank support is flat, the liquid reserve tank is placed on the upper portion of liquid reserve tank support, and liquid reserve tank support level is placed on the upper portion of second lifting unit, along with the rising or the decline of second lifting unit, drives liquid reserve tank support and places the liquid reserve tank on the liquid reserve tank support and rises or descend.
The top of the liquid storage tank is provided with a liquid storage tank top cover used for sealing the liquid storage tank body, a liquid storage tank air filter is arranged on the liquid storage tank top cover, and gas entering and exiting the liquid storage tank is filtered to prevent pollution.
Particularly, the middle lower part of the liquid storage tank is provided with a liquid storage tank liquid inlet/outlet hole, and the size of the liquid storage tank liquid inlet/outlet hole is matched with that of the connecting pipeline.
The first lifting assembly and the second lifting assembly are selected from a lifting platform or a lifter, and preferably from the lifting platform.
In particular, the lifting platform is an external force mechanical system such as an electric lifting platform, a hydraulic lifting platform, an air pressure lifting platform and the like.
The lifting assembly is arranged on the ground or in a laboratory bench or a sterile workshop. The lifting height, the lifting speed, the lifting time or the lifting time of the lifting component are controlled automatically or manually. Computer application instructions are typically employed to precisely control or manually manipulate the raising and lowering of the lift assembly in emergency situations.
The lifting control assembly is connected with the lifting assembly through one end of a wire, the other end of the wire is connected with a computer, an instruction signal of the computer is transmitted to the lifting control assembly through the wire, and the lifting control assembly controls the lifting assembly to ascend, descend or keep the height unchanged through the wire.
The lifting control assembly comprises a controller, a computer and a lead. The controller is a device known in the art that can control the raising or lowering of the lift platform or lift, such as an electric switch.
Compared with the prior art, the intermittent immersion bioreactor has the following advantages and benefits:
1. the bioreactor has a simple structure, the tissue culture assembly and the liquid assembly are communicated through a pipeline to form a closed system, and the tissue culture assembly and the liquid assembly are integrally used to reduce contamination probability during operation; the method reduces the production cost of plant tissue culture, improves the multiplication coefficient and reduces the production labor operation (mainly inoculating plant materials).
2. The bioreactor is easy to operate, the tissue culture component and the liquid component are arranged on the lifting platform (plate), are separated from the power system, are independent from each other and easy to operate, the culture conditions of plant tissue culture influenced by the time and frequency of immersing plant materials in a liquid culture medium can be accurately controlled, and the efficiency of the plant tissue culture is improved;
3. the power system for controlling the lifting component to lift in the bioreactor can simultaneously operate a plurality of lifting components, thereby controlling the lifting of a plurality of tissue culture components and liquid components and improving the tissue culture efficiency of plants.
4. In the bioreactor, because the culture solution flows from a high position to a low position under the driving of the self gravity of the culture solution, a plurality of tissue culture assemblies and liquid assemblies with the same or different sizes can be arranged on the same lifting assembly, and the culture treatment of a plurality of or different plant tissues is realized simultaneously.
5. In the bioreactor, a plurality of tissue culture assemblies and liquid assemblies which are different or identical in tissue culture are used, so that unified management is facilitated, and the bioreactor is suitable for standard production.
Drawings
FIG. 1 is a schematic view of the structure of a temporary immersion bioreactor according to example 1 of the present invention;
FIG. 2 is a schematic view of the structure of the culture assembly of the present invention;
FIG. 2A is a cross-sectional view taken along line A-A of FIG. 2;
FIG. 3 is a schematic view of a fluid module according to the present invention;
FIG. 4 is a schematic view showing the operation state of the batch immersion bioreactor according to the present invention in which the liquid storage tank injects the culture solution into the culture tank;
FIG. 5 is a schematic view showing the operation of discharging the culture solution from the culture tank to the liquid storage tank in the temporary immersion bioreactor of the present invention;
FIG. 6 is a schematic view of the structure of a temporary immersion bioreactor according to example 1A of the present invention;
FIG. 6A is a schematic view of the structure of a temporary immersion bioreactor according to example 1B of the present invention;
FIG. 6B is a schematic structural view of a support plate according to the present invention;
FIG. 7 is a schematic view of the structure of a temporary immersion bioreactor in example 2 of the present invention;
FIG. 8 is a schematic view of the structure of a temporary immersion bioreactor according to example 3 of the present invention.
Description of the reference numerals
1. A culture assembly; 11. an incubator; 12. an incubator support; 13. a culture box cover; 14. an air filter; 15. an incubator liquid inlet/outlet hole; 16. an inner support box; 17. a liquid flow aperture; 18. a support plate; 2. a liquid component; 21. a liquid storage tank; 22. a reservoir holder; 23. a top cover of the liquid storage tank; 24. liquid inlet/outlet holes of the liquid storage tank; 3. a first lifting assembly; 4. a second lifting assembly; 5. and connecting the pipelines.
Detailed Description
The advantageous effects of the present invention will be further described below by way of specific embodiments, which are merely exemplary and do not set any limit to the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Example 1
As shown in fig. 1, the intermittent immersion bioreactor of the present invention comprises a tissue culture assembly 1, a liquid assembly 2, a first lifting assembly 3 for raising or lowering the tissue culture assembly, and a second lifting assembly 4 for raising or lowering the liquid assembly, wherein the tissue culture assembly is installed on the upper portion of the first lifting assembly; the liquid assembly is arranged at the upper part of the second lifting assembly. The tissue culture assembly is communicated with the liquid assembly through a connecting pipeline 5, and culture solution stored in the liquid assembly is conveyed into the tissue culture assembly through rising or lowering of the first lifting assembly and the second lifting assembly, or the culture solution in the tissue culture assembly is discharged and conveyed into the liquid assembly, and intermittent immersion treatment is carried out on plant tissues in the tissue culture assembly.
Or keeping the height of the first lifting component unchanged, and injecting the culture solution in the liquid component into the tissue culture component through the lifting of the second lifting component; the culture solution in the tissue culture assembly flows into the liquid assembly through the lowering of the second lifting assembly; keeping the height of the second lifting assembly unchanged, and injecting the culture solution in the liquid assembly into the tissue culture assembly through the reduction of the first lifting assembly; through the rising of second lifting unit for the culture solution in the group banks up the subassembly with earth flows into liquid subassembly, realizes that the culture solution is to the intermittent type submergence of waiting to handle the material in the group banks up the subassembly with earth.
As shown in fig. 1 and 2, the tissue culture assembly 1 is used for placing plant materials to be subjected to tissue culture treatment, and the plant materials are subjected to tissue culture in the tissue culture assembly, and comprises an incubator 11 and an incubator support 12 for placing and fixing the incubator, wherein the incubator support plate is flat, the incubator is placed on the upper portion of the incubator support plate, the incubator support is horizontally placed on the upper portion of the first lifting assembly, and the incubator support and the incubator placed on the upper portion of the incubator support are driven to ascend or descend along with the ascending or descending of the first lifting assembly.
The top of the incubator is provided with an incubator cover 13 for closing the incubator body, the incubator cover is provided with an air filter 14 for eliminating pressure difference in the incubator caused by liquid level lifting, air enters and exits the incubator and filters gas entering and exiting the incubator to prevent pollution; as shown in FIG. 2, the lower middle part of the incubator is provided with an incubator liquid inlet/outlet hole 15 having a size matching that of the connecting pipe 5, the connecting pipe is fixedly connected and sealed with the liquid inlet/outlet hole, and the culture liquid is injected into the incubator or discharged from the incubator. The liquid inlet/outlet holes are usually formed near the bottom of the incubator to facilitate the complete discharge of the culture solution from the incubator.
As shown in fig. 2A, a tissue culture inner support box 16 is further disposed inside the incubator, and is used for placing plant materials to be treated, i.e., providing a support surface (fixed position) for the plant materials, and separating from the liquid surface when necessary. The bottom of the inner supporting box can also be paved with water-absorbing fiber materials, thereby having the effect of infiltrating plant materials. The height of the inner support box is 1/2-2/3 of the height of the incubator, the size of the horizontal section of the inner support box is matched with that of the horizontal section of the incubator, the top of the inner support box is clamped and fixed with the top of the incubator and is equal to the top of the incubator in the same horizontal plane; the bottom of the inner support box is spaced from the bottom of the incubator by a certain distance to form a culture solution storage space; the bottom of the inner support box is uniformly provided with a plurality of liquid flowing holes 17 for culture solution to flow freely and enter and exit the inner support box freely, so that the plant material to be treated is immersed intermittently.
As shown in fig. 1 and 3, the liquid assembly 2 is used for storing liquid culture medium and comprises a liquid storage tank 21 and a liquid storage tank support 22 for placing and fixing the liquid storage tank, the liquid storage tank support is flat, the liquid storage tank is placed on the upper portion of the liquid storage tank support, the liquid storage tank support is horizontally placed on the upper portion of the second lifting assembly, and the second lifting assembly ascends or descends to drive the liquid storage tank support and the liquid storage tank placed on the liquid storage tank support to ascend or descend.
A liquid storage tank top cover 23 used for sealing the liquid storage tank body is arranged at the top of the liquid storage tank, and a liquid storage tank air filter 14 is arranged on the liquid storage tank top cover and used for eliminating pressure difference in the liquid storage tank caused by lifting of the liquid level, utilizing air to enter and exit the liquid storage tank, and filtering gas entering and exiting the liquid storage tank to prevent pollution; liquid storage tank inlet/outlet 24 is formed in the middle lower portion of the liquid storage tank, the size of the liquid storage tank inlet/outlet is matched with the size of the connecting pipeline 5, the connecting pipeline and the liquid storage tank inlet/outlet are fixedly connected and sealed, and culture solution is injected into the liquid storage tank or discharged out of the liquid storage tank. The liquid inlet/outlet hole of the liquid storage tank is usually arranged at a position close to the bottom of the liquid storage tank, so that the culture liquid in the liquid storage tank can be completely discharged.
The culture box of the tissue culture assembly is communicated with the liquid storage box of the liquid assembly through a connecting pipeline 5, when the liquid level of the culture solution in the liquid storage box is higher than the liquid level in the culture box, the culture solution in the liquid storage box flows into the culture box and submerges plant materials placed on the bottom of an inner support box in the culture box; when the liquid level in the incubator is higher than the liquid level in the liquid storage tank, the culture solution in the incubator flows into the liquid storage tank, and the plant material originally immersed in the liquid culture medium is separated from the liquid culture medium immediately.
The plant material to be treated according to the invention can be placed directly on the bottom of the incubator or on a mat on the bottom.
The first lifting component and the second lifting component select a lifting platform, preferably an electric lifting platform.
The lifting assembly is arranged on the ground or in a laboratory bench or a sterile workshop, the specific embodiment of the invention is partially described by taking an electric lifting platform as an example, and other mechanical systems such as a hydraulic lifting platform, an air pressure lifting platform or a manual lifting platform are suitable for the invention besides the electric lifting platform.
The electric lifting platform controls the lifting or descending of the lifting platform through a lifting control component (not shown in the figure, such as an electric switch), the lifting control component is connected with a computer through a lead, a control signal of the computer is transmitted to the lifting control component, and the lifting or descending of the lifting component is controlled or the height is kept unchanged.
First lifting unit can fixed height, the first lifting unit of lift control subassembly control does not rise, does not reduce, keep first lifting unit height unchangeable, only control second lifting unit and rise or reduce, perhaps lifting unit control second lifting unit does not rise/reduce, keep second lifting unit height unchangeable, after the fixed certain height of second lifting unit, only control first lifting unit and reduce or rise for the culture solution in the liquid reserve tank of liquid subassembly injects the incubator or injects the culture solution in the incubator into the liquid reserve tank.
The lifting height, lifting speed and lifting time of the first and second lifting assemblies are precisely controlled by a lifting control assembly (not shown). The lifting control assembly receives computer program instructions or is manually operated. In the embodiment of the present invention, the lifting control component selects the electric control switch, and besides the electric switch, other devices known in the art for controlling the lifting platform or the lifter to ascend or descend are suitable for the lifting control component of the present invention.
The lift control assembly includes: computer, controller and the wire that connects computer and controller. According to the operating conditions for processing plant materials, corresponding parameters are input into a computer, and the lifting direction, distance (height), speed, stopping time and the like of the lifting platform are controlled by computer software. The computer signal is sent to the lifting platform with the motor through the controller so as to complete the instruction.
The lifting control assembly is connected with a computer through a wire, the lifting height, the lifting speed and the lifting initial time of the lifting assembly lifting platform are accurately controlled according to corresponding control parameters in the computer, the immersion time of a liquid culture medium is accurately controlled, and accurate tissue culture treatment of plant materials is realized.
The working principle of the intermittent immersion bioreactor is as follows:
1. as shown in fig. 1, the first lifting assembly and the second lifting assembly are adjusted to be at the same height, the incubator support and the liquid storage tank support are respectively and horizontally placed on the first lifting platform and the second lifting platform, the incubator is placed and fixed on the incubator support, and the liquid storage tank is placed and fixed on the liquid storage tank support; the culture box is communicated with the liquid storage box through a connecting pipeline;
2. adjusting the relative heights of the first lifting component and the second lifting component through the lifting control component, wherein along with the fact that the height of the second lifting component is higher than that of the first lifting component, when the liquid level in the incubator is lower than that of the culture solution in the liquid storage tank, the culture solution in the liquid storage tank flows into the incubator, and plant tissues to be treated in the incubator are immersed until the liquid levels in the incubator and the liquid storage tank are consistent, as shown in figure 4;
3. after the plant tissue is immersed for a certain time, the relative height of the first lifting component and the second lifting component is adjusted through the lifting control component, along with the fact that the height of the second lifting component is lower than that of the first lifting component, when the liquid level in the incubator is higher than that of the culture liquid in the liquid storage tank, the culture liquid in the incubator flows into the liquid storage tank, the plant tissue immersed in the liquid culture medium is not immersed in the liquid culture medium, and the immersion treatment of the plant tissue is stopped, as shown in fig. 5. The plant tissue in the incubator is separated from the liquid medium, thereby achieving intermittent immersion of the plant tissue.
When the intermittent immersion bioreactor is used for tissue culture, the culture box and the inner support box of the tissue culture assembly of the bioreactor and the liquid storage box of the liquid assembly are sterilized at high temperature and then are placed on a superclean workbench. Placing plant material to be cultured in the inner supporting box under aseptic condition, pouring appropriate amount of liquid culture medium into the liquid storage tank, and covering the container cover. The tissue culture assembly and the liquid assembly are respectively arranged on the first group of lifting platform and the second group of lifting platform, and the rubber tube connected between the first group of lifting platform and the second group of lifting platform is kept in a proper length.
Tissue culture subassembly and liquid subassembly place and fix the back, the input is used for controlling first, the relative lifting motion's of second elevating platform subassembly corresponding to count the machine program of formula. Along with the decline or the rising of first, second lifting unit for the height of the group culture subassembly on the second lifting unit is higher than the height of group culture subassembly on the first lifting unit, and liquid medium flows to the incubator from the liquid reserve tank because of the effect of gravity, thereby accomplishes soaking to plant material. On the contrary, along with the ascending or descending of the first lifting component and the second lifting component, the height of the liquid component on the second lifting component is lower than that of the tissue culture component on the first lifting component, and the liquid reversely flows into the liquid storage tank, so that the plant material is separated from the liquid. The time, speed and number of times of the repetition of the process are all controlled by computer instructions. The movement of the first lifting platform (group) or the movement of the two groups of lifting platforms can be controlled to change the relative heights of the liquid storage box and the culture box, so that the liquid flow direction can be controlled.
Example 1A
The same as example 1 was used except that the tissue culture assembly contained no internal support box in the incubator and the plant material to be treated was placed directly on the bottom of the incubator. As shown in fig. 6.
Example 1B
In addition to the inner support box, a support plate 18 may be provided inside the incubator instead of the inner support box, the support plate being provided at a lower middle portion of the incubator and having a shape and a size corresponding to those of the horizontal cross section of the incubator, the support plate being provided at a height spaced from the bottom of the incubator, for example, a ratio of the height from the bottom of the incubator to the height of the incubator is 1/3 to 1/2. As shown in FIG. 6B, a plurality of liquid flowing holes 17 are uniformly formed in the supporting plate, so that the culture solution can freely flow and freely pass through the supporting plate, and the plant material to be treated can be intermittently immersed.
The same as in example 1 was used except that the inner support box in the incubator of the tissue culture assembly was replaced by a support plate and the plant material to be treated was placed directly on the support plate of the incubator. As in fig. 6A.
Example 2
As shown in fig. 7, a plurality of (e.g., 2) incubators are placed on the incubator support 12; a plurality of (e.g., 2) liquid tanks are placed on the liquid tank support plate 22; the number of the culture boxes is consistent with that of the liquid storage boxes, the culture boxes correspond to the liquid storage boxes to form a plurality of groups (for example, 2 groups) of culture-liquid storage boxes, and the culture boxes and the liquid storage boxes in each group of culture-liquid storage boxes are communicated through connecting pipelines.
As shown in fig. 6, the first lifting assembly selects 1 lifting platform (preferably an electric lifting platform); the second lifting assembly selects 1 lifting platform (preferably an electric lifting platform). The incubator bracket 12 is horizontally placed and fixed on 2 lifting platforms of the first lifting component; the tank support 22 is horizontally placed on 2 lifting platforms of the second lifting assembly.
Example 3
As shown in FIG. 8, a plurality of (e.g., 4) incubators are placed on the incubator support plate 12; a plurality of (e.g., 4) liquid tanks are placed on the liquid tank support plate 22; the number of the culture boxes is consistent with that of the liquid storage boxes, the culture boxes correspond to the liquid storage boxes to form a plurality of groups (for example, 4 groups) of culture-liquid storage boxes, and the culture boxes and the liquid storage boxes in each group of culture-liquid storage boxes are communicated through connecting pipelines.
As shown in fig. 7, the first lifting assembly selects a plurality (e.g., 2) of lifting platforms (preferably electric lifting platforms); the second lifting assembly selects a plurality (e.g., 2) of lifting platforms (preferably motorized lifting platforms). The incubator bracket 12 is horizontally placed and fixed on 2 lifting platforms of the first lifting component; the tank support 22 is horizontally placed on 2 lifting platforms of the second lifting assembly.
A plurality of lifting platforms through the first lifting component of lift control subassembly control rise simultaneously or descend or through a plurality of lifting platforms of lift control subassembly control second lifting component rise simultaneously or descend, perhaps control the first lifting component of lift control subassembly and rise simultaneously and control the decline of second lifting component, perhaps control the decline of first lifting component through the lift control subassembly and control the second lifting component simultaneously and rise, make the liquid level height in incubator and the liquid reserve tank inconsistent, make the culture solution flow between incubator and liquid reserve tank, the realization is to the intermittent type submergence of the pending plant tissue in the incubator.
Claims (10)
1. A potential energy driven intermittent immersion bioreactor comprises a tissue culture component and a liquid component, and is characterized by also comprising a first lifting component and a second lifting component, wherein,
the tissue culture assembly is used for placing plant materials to be subjected to tissue culture treatment, and the plant materials are subjected to tissue culture in the tissue culture assembly; the liquid component is used for storing liquid culture medium; the tissue culture assembly is communicated with the liquid assembly through a connecting pipeline;
the first lifting assembly is used for lifting or lowering the tissue culture assembly; the second lifting assembly is used for lifting or lowering the liquid assembly, and along with the lowering of the first lifting assembly or the lifting of the second lifting assembly, the culture solution in the liquid assembly is injected into the tissue culture assembly; along with the rising of first lifting unit or the reduction of second lifting unit for the culture solution in the group banks up the subassembly with earth and flows into the liquid subassembly, realizes that the culture solution is to the intermittent type submergence of the material of treating in the group banks up the subassembly with earth and handle.
2. The bioreactor as claimed in claim 1, wherein the tissue culture assembly is communicated with the liquid assembly through a connecting pipeline, and the culture solution in the liquid assembly is injected into the tissue culture assembly through the connecting pipeline by the lowering of the first lifting assembly or the raising of the second lifting assembly; through the rising of first elevating system or the reduction of second elevating system, the culture solution of group in the subassembly of cultivateing discharges into in the liquid subassembly through connecting tube, carries out intermittent type submergence processing to the plant tissue of group in the subassembly of cultivateing.
3. The bioreactor of claim 1, wherein the tissue culture assembly comprises an incubator and an incubator support, wherein the incubator is disposed and fixed on an upper portion of the incubator support.
4. The bioreactor of claim 3, wherein the incubator support is formed in a flat plate shape, the incubator is placed on an upper portion of the incubator support plate, the incubator support is horizontally placed on an upper portion of the first elevating unit, and the incubator support and the incubator placed on the upper portion of the incubator support are moved up and down by the first elevating unit in accordance with the elevation or lowering of the first elevating unit.
5. The bioreactor of claim 3, wherein the top of the incubator is provided with an incubator cover for closing the incubator body, and the incubator cover is provided with an air filter for filtering air entering and exiting the incubator to prevent contamination.
6. The bioreactor of claim 3, wherein the lower part of the incubator is provided with an incubator inlet/outlet hole, and the size of the incubator inlet/outlet hole is matched with that of the connecting pipeline.
7. The bioreactor of claim 1, wherein the fluid module comprises a reservoir and a reservoir holder, wherein the reservoir is positioned and secured on an upper portion of the reservoir holder.
8. The bioreactor of claim 7, wherein the tank holder is formed in a flat plate shape, the tank is placed on an upper portion of the tank holder plate, the tank holder is horizontally placed on an upper portion of the second elevating unit, and the tank holder and the tank placed on the upper portion of the tank holder are moved up and down by the second elevating unit as the second elevating unit is elevated or lowered.
9. The bioreactor of claim 7, wherein the top of the tank is provided with a tank top cover for sealing the tank body, and the top cover is provided with an air filter for filtering the gas entering and exiting the tank to prevent contamination.
10. The bioreactor of claim 7, wherein the lower portion of the tank is provided with a tank inlet/outlet hole having a size matching the size of the connection pipe.
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| CN201811545726.8A CN110896852A (en) | 2018-12-18 | 2018-12-18 | Potential energy driven intermittent immersion type bioreactor |
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