Simple device for culturing adventitious roots of medicinal plants

Abstract

The utility model discloses a simple medicinal plant adventitious root culture device, the utility model discloses 10L transparent high borosilicate glass bottle is adopted, compared with traditional triangular flask has larger volume, only needs inoculation, can reduce the pollution risk, replaces the mechanical vibration of traditional shaking bed through the airflow of air pump and stirs, and the design of no shaking bed dependence makes the equipment occupied space reduce, can reduce the purchase cost, and the maintenance cost of shaking bed is saved, and laboratory culture cost is reduced significantly, and the medium and device itself can be connected common vertical autoclave sterilization through quick connector, and the design of quick connector makes the medium replacement efficiency promote, and operating personnel can carry out aseptic supplement, replace and sampling operation to the medium in the culture process, and air distributor makes the uniformity of dissolved oxygen improve significantly, and solves the problem of adventitious root blockage, and the culture stability improves significantly, and integrated sterilization scheme makes the preparation time shorten in advance, and avoids the secondary pollution risk after the sterilization of part alone.

Description

Technical Field

[0001] This utility model relates to the field of plant tissue culture technology, and more specifically, to a simple adventitious root culture device for medicinal plants. Background Technology

[0002] As an important source of active ingredients in natural drugs, the large-scale laboratory cultivation technology of adventitious roots of medicinal plants has always been a research hotspot in the field of plant biotechnology. Currently, the mainstream cultivation methods are mainly divided into two categories: Erlenmeyer flask shaking culture and small-scale bioreactor culture, but both have significant technical bottlenecks in practical applications.

[0003] Erlenmeyer flask culture, as the most traditional culture method, has inherent drawbacks that are difficult to overcome: 1. Erlenmeyer flask culture is small in volume, and the accumulation of adventitious roots requires multiple manual inoculations, increasing the risk of contamination; multi-flask culture is also cumbersome. 2. The use of shakers occupies a large amount of space. 3. Purchasing shakers requires a significant investment.

[0004] Although small-scale bioreactors can achieve a certain scale of cultivation, the cost of the bioreactor itself and its supporting equipment is high, and the operation requires a certain level of expertise. Moreover, the operation of changing the culture medium during the adventitious root cultivation process in small-scale bioreactors is quite difficult.

[0005] Some improvements have been attempted in existing technologies, such as replacing shake flasks with larger-volume glass containers, but still relying on shakers for power; or simplifying the reactor structure to eliminate the stirring system, but due to insufficient dissolved oxygen, adventitious root growth is slow, the total biomass increase is reduced, and the adventitious root biomass obtained per unit time or per unit of resource input is lower than normal. These improvements have not fundamentally solved the three-in-one problem of "high pollution risk, complex operation, and high cost." Therefore, there is an urgent need to develop a new cultivation device that balances high-efficiency cultivation, simple operation, and controllable cost. Utility Model Content

[0006] This invention provides a simple device for culturing adventitious roots of medicinal plants, aiming to solve the problems of high pollution risk, large space occupation, high cost, cumbersome operation, and difficulty in changing culture medium in the existing technology for culturing adventitious roots of medicinal plants in the laboratory.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: a simple medicinal plant adventitious root culture device, comprising a sterile bottle, a multi-port cap at the bottle mouth, an air inlet pipe and an exhaust pipe inserted into the multi-port cap, one end of the air inlet pipe and the exhaust pipe communicating with the internal cavity of the sterile bottle, and the other end of the air inlet pipe and the exhaust pipe communicating with the external space, one end of the air inlet pipe connected to an air distributor located inside the sterile bottle, the other end of the air inlet pipe connected to an air pump located in the external space, air filters provided at the ends of the air inlet pipe and the exhaust pipe, and several quick-connect holes provided on the side wall of the sterile bottle along its height direction, with quick connectors provided at the quick-connect holes.

[0008] Furthermore, the sterile bottle is a 10L transparent high borosilicate glass bottle.

[0009] Furthermore, the multi-port bottle cap is selected as a two-port bottle cap, and the two-port bottle cap has an air inlet connected to the air inlet pipe and an exhaust outlet connected to the exhaust pipe.

[0010] Furthermore, a high-temperature resistant rubber gasket is provided between the mouth of the sterile bottle and the multi-port bottle cap.

[0011] Furthermore, the air intake pipe is equipped with a plastic clip.

[0012] Furthermore, the quick connector is selected as a φ8mm pagoda head quick connector.

[0013] Furthermore, there are two quick-connect holes and two quick-connect connectors, one of which is located at the upper end of the sterile bottle and the other is located at the lower end of the sterile bottle.

[0014] Furthermore, the air filter is selected as a 0.2μm microporous membrane filter.

[0015] Furthermore, the multi-port bottle cap is provided with a connecting pipe, which is connected to the air inlet pipe. The air distributor is located at the port of the connecting pipe, and the air inlet pipe, the connecting pipe, and the air distributor are arranged sequentially from top to bottom.

[0016] Furthermore, the quick connector located at the upper end of the sterile bottle is equipped with a peristaltic pump.

[0017] Compared with existing technologies, the beneficial effects of this utility model are as follows: This utility model uses a 10L transparent high borosilicate glass bottle, which has a larger volume than the traditional triangular shake flask. It only requires one inoculation, which can reduce the risk of contamination. The transparent bottle allows for direct observation of the culture status, further improving the ease of operation. The airflow stirring by the air pump replaces the mechanical shaking of the traditional shaker. The shaker-free design reduces the space occupied by the equipment, which can reduce procurement costs and eliminate shaker maintenance costs, significantly reducing laboratory culture costs. The culture medium and the device itself can be sterilized by connecting to a common 80L vertical autoclave in the laboratory via quick connectors. The quick connector design improves the efficiency of culture medium replacement, allowing operators to perform aseptic replenishment, replacement, and sampling of the culture medium during the culture process. The 30μm air distributor significantly improves dissolved oxygen uniformity and solves the problem of adventitious root blockage, significantly improving culture stability. The integrated sterilization scheme shortens the preparation time, avoids the risk of secondary contamination after individual component sterilization, and has a higher operational error tolerance. Attached Figure Description

[0018] To more clearly illustrate the technical solutions of the embodiments of this utility model, the drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this utility model and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained from these drawings without creative effort.

[0019] Figure 1 This is a schematic diagram of the structure of this utility model;

[0020] Figure 2 This is a schematic diagram of aseptic airflow during the cultivation process of this utility model;

[0021] Figure 3 This is a schematic diagram of the flow of the culture medium during the replenishment of the culture medium according to this utility model;

[0022] Figure 4 This is a schematic diagram of the flow of the culture medium during the discharge of the culture medium according to this utility model.

[0023] Explanation of main component symbols

[0024] 1. Sterile bottle; 2. Multi-port cap; 21. Air inlet; 22. Exit outlet; 3. Air inlet pipe; 4. Exit pipe; 5. Air filter; 6. Air distributor; 7. Quick connector; 8. Air pump; 9. Plastic clip; 10. Connecting pipe. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this utility model, not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model. Therefore, the following detailed description of the embodiments of this utility model provided in the accompanying drawings is not intended to limit the scope of the claimed utility model, but merely represents selected embodiments of this utility model. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model.

[0026] In this utility model, unless otherwise explicitly specified and limited, the terms "installation," "connection," "joining," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0027] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on top" of the second feature includes the first feature directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature includes the first feature directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0028] Example

[0029] like Figures 1-4As shown, this utility model discloses a simple adventitious root culture device for medicinal plants, including a sterile bottle 1. The sterile bottle 1 has a multi-port cap 2 at its mouth. An air inlet pipe 3 and an exhaust pipe 4 are inserted into the multi-port cap 2. One end of the air inlet pipe 3 and the exhaust pipe 4 are connected to the internal cavity of the sterile bottle 1, and the other end of the air inlet pipe 3 and the exhaust pipe 4 are connected to the external space. One end of the air inlet pipe 3 is connected to an air distributor 6, which is located inside the sterile bottle 1. The other end of the air inlet pipe 3 is connected to an air pump 8, which is located in the external space. Air filters 5 are provided at the ports of the air inlet pipe 3 and the exhaust pipe 4. Several quick-connect holes are opened on the side wall of the sterile bottle 1 along its height direction, and quick-connect connectors 7 are provided at the quick-connect holes.

[0030] This invention uses a multi-port bottle cap 2 to make the sterile bottle 1 a closed, sterile culture container; the air inlet pipe 3 can deliver sterile air into the sterile bottle 1 to provide sufficient dissolved oxygen for the culture of adventitious roots, while the tail gas pipe 4 at the other end discharges the tail gas to balance the pressure inside the bottle; the air distributor 6 has both efficient dissolved oxygen and anti-clogging functions, which can evenly disperse air and prevent adventitious roots from clogging the distribution holes; the air pump 8 introduces sterile air to form an airflow that replaces the mechanical shaking of the traditional shaker, realizing a culture mode without the dependence on a shaker; the air filter 5 can prevent microorganisms from entering the sterile bottle 1 through the air inlet pipe 3 and the tail gas pipe 4, providing a sterile environment for the culture of adventitious roots; the quick-connect hole allows the culture medium to pass through, and the quick-connect joint 7 can control the inflow and outflow of the culture medium.

[0031] Among them, sterile bottle 1 is a 10L transparent high borosilicate glass bottle. The use of transparent high borosilicate glass bottle makes it easy for operators to observe the condition inside the bottle.

[0032] Among them, the multi-port bottle cap 2 is a two-port bottle cap. The two-port bottle cap has an air inlet 21 connected to the air inlet pipe 3 and an exhaust port 22 connected to the exhaust pipe 4. The two-port bottle cap integrates air intake regulation and exhaust gas emission, realizing the overall control of multiple functions of the culture system by a single bottle cap.

[0033] Among them, a high-temperature resistant rubber gasket is provided between the mouth of the sterile bottle 1 and the multi-port bottle cap 2, which can ensure the airtightness of the sterile bottle 1.

[0034] The intake pipe 3 is equipped with a plastic clip 9, which can control the flow of gas inside the intake pipe 3.

[0035] Among them, quick connector 7 is selected as φ8mm pagoda head quick connector 7, and the pagoda head quick connector hole can facilitate the connection between the quick connector hole and the external pipe.

[0036] There are two quick-connect holes and two quick-connect connectors 7. One quick-connect hole is located at the upper end of the sterile bottle 1, and the other quick-connect hole is located at the lower end of the sterile bottle 1. The culture medium in the sterile bottle 1 can flow out from the quick-connect hole and quick-connect connector 7 located at the lower end of the sterile bottle 1 under the action of gravity. The operator can transfer the culture medium into the sterile bottle 1 through the quick-connect hole and quick-connect connector 7 located at the upper end of the sterile bottle 1.

[0037] Among them, the air filter 5 is a 0.2μm microporous membrane filter, which can effectively prevent microorganisms from entering the sterile bottle 1.

[0038] The multi-port bottle cap 2 is provided with a connecting pipe 10, which is connected to the air inlet pipe 3. The air distributor 6 is located at the port of the connecting pipe 10. The air inlet pipe 3, the connecting pipe 10 and the air distributor 6 are arranged in sequence from top to bottom.

[0039] The quick connector 7 located at the upper end of the sterile bottle 1 is equipped with a peristaltic pump, which can work with the quick connector 7 and quick connector hole to deliver culture medium to the sterile bottle 1.

[0040] Device assembly: Install the multi-port bottle cap 2 onto the mouth of the sterile bottle 1 and seal it with a high-temperature resistant rubber gasket. A 30μm air distributor 6 is installed at the lower end of the connecting pipe 10 integrated on the multi-port bottle cap 2, and the upper end is connected to the air inlet pipe 3. The air pump 8 is connected to the air distributor 6 via a silicone hose, a 0.2μm air filter 5, the air inlet pipe 3, the air inlet 21 of the multi-port bottle cap 2, and the connecting pipe 10. The exhaust port 22 of the multi-port bottle cap 2 is connected to another 0.2μm air filter 5 via an exhaust pipe 4, forming a complete sterile airflow circulation system. Quick-connect holes and quick-connect connectors 7 can adapt to different culture medium operation requirements.

[0041] Sterilization: After filling the culture medium into the sterile bottle 1, tighten the multi-port cap 2, clamp the air inlet tube 3 with the plastic clip 9, and introduce external steam through the quick connector 7. Perform high-pressure steam sterilization at 121°C for 20 minutes. No need to disassemble parts for separate sterilization, shortening the preparation time. If it is necessary to replenish, replace or sample the culture medium during the culture process, the sterile container and the culture medium storage bottle can be connected to the quick connector 7 and the external silicone tube for joint sterilization before sterilization to avoid secondary assembly contamination. The sterile container and the culture medium storage bottle can also be sterilized separately and then aseptically connected directly on the ultra-clean workbench during the culture process.

[0042] Inoculation: After cooling, open the multi-port bottle cap 2 in the ultra-clean workbench to complete aseptic inoculation. The integrated design of the multi-port bottle cap 2 reduces the exposure time of the opening and lowers the risk of contamination.

[0043] Incubation: After tightening the multi-port bottle cap 2, place the bottle in a suitable temperature environment and turn on the air pump 8 to achieve a shaker-free incubation mode. Airflow agitation replaces the mechanical shaking of a traditional shaker, saving equipment space. The 30μm air distributor 6 evenly releases sterile air, improving dissolved oxygen uniformity. The exhaust gas is discharged through the exhaust pipe 4 to balance the pressure and ensure the stability of the incubation system.

[0044] Culture medium operation: When culture medium needs to be replenished, the peristaltic pump is connected to the quick connector 7 at the upper end of the sterile bottle 1 to deliver the culture medium into the sterile bottle 1; when culture medium needs to be discharged, the culture medium can flow out of the sterile bottle 1 under the action of gravity through the quick connector hole and quick connector 7 at the lower end of the sterile bottle 1; when no operation is required, the quick connector 7 is kept closed, and the whole process realizes a closed loop of sterile operation.

[0045] The above are merely preferred embodiments of this utility model and are not intended to limit the scope of this utility model. Various modifications and variations can be made to this utility model by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of this utility model should be included within the protection scope of this utility model.

Claims

1. A simple device for cultivating adventitious roots of medicinal plants, characterized in that, The device includes a sterile bottle with a multi-port cap at its opening. An air inlet pipe and an air outlet pipe are inserted into the multi-port cap. One end of each pipe communicates with the internal cavity of the sterile bottle, while the other end communicates with the external space. One end of the air inlet pipe is connected to an air distributor located inside the sterile bottle, and the other end is connected to an air pump located in the external space. Air filters are provided at both ends of the air inlet and air outlet pipes. Several quick-connect holes are formed on the side wall of the sterile bottle along its height, and quick-connect fittings are provided at these holes.

2. The simple adventitious root culture device for medicinal plants according to claim 1, characterized in that; The sterile bottle is a 10L transparent high borosilicate glass bottle.

3. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that; The multi-port bottle cap is selected as a two-port bottle cap, which has an air inlet connected to the air inlet pipe and an exhaust outlet connected to the exhaust pipe.

4. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that; A high-temperature resistant rubber gasket is provided between the mouth of the sterile bottle and the multi-port bottle cap.

5. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that; The air intake pipe is equipped with a plastic clip.

6. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that: The quick connector selected is a φ8mm pagoda head quick connector.

7. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that: There are two quick-connect holes and two quick-connect connectors, one of which is located at the upper end of the sterile bottle and the other at the lower end of the sterile bottle.

8. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that: The air filter is selected as a 0.2μm microporous membrane filter.

9. A simple medicinal plant adventitious root culture device according to claim 1, characterized in that; The multi-port bottle cap is provided with a connecting pipe, which is connected to the air inlet pipe. The air distributor is located at the port of the connecting pipe, and the air inlet pipe, the connecting pipe, and the air distributor are arranged sequentially from top to bottom.

10. A simple medicinal plant adventitious root culture device according to claim 7, characterized in that; The quick connector located at the upper end of the sterile bottle is equipped with a peristaltic pump.

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