• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Reactor system for the in vitro cultivation of plant material, kit for transforming a receptacle into a reactor suitable for said system, and method for the in vitro cultivation of plant material using said reactor system. It comprises a receptacle (1) for the cultivation of plant material (11), a lid (2) to close an opening (3) of said receptacle (1), and means (8, 9) to allow entry and/or exit gas of said receptacle (1), and is characterized by the fact that it comprises a container (4) to receive a liquid culture medium inside the receptacle (1), and means (6, 7) to support the material (11) vegetable on said receiving container (4), where said container (4) is attached to the lid (2) of the receptacle (1) in a position suitable for use, said lid (2) including at least one conduit (5) input and/or output medium that communicates with the container (4) to be able to fill and/or empty said container (4) through said conduit (5). (Machine-translation by Google Translate, not legally binding)
    公开号:ES2763637A1
    申请号:ES201831164
    申请日:2018-11-29
    公开日:2020-05-29
    发明作者:Sanjuan Ramon J Dolcet；Morales Carlos Rolando Mendoza
    申请人:Institut Recerca i Tecnologia Agroalimentaries IRTA；
    IPC主号:
    专利说明:

    [0004] The present invention relates to a reactor system for clonal propagation of plants comprising a receptacle for the in vitro cultivation of plant material. It also refers to a kit suitable for transforming a receptacle into an in vitro culture reactor of plant material suitable for said system, and to a method for in vitro culture of plant material using said reactor system.
    [0006] Background of the Invention
    [0008] Reactor systems for the in vitro culture of plant material are known, such as for example Plantform ® brand or Rita ® brand reactor systems, which comprise a receptacle provided with a tray to support the plant material, a cover for closing an opening of the receptacle and a plurality of air injection ducts with which temporary pumping of a liquid culture medium stored at the bottom of the culture receptacle is achieved. The temporary immersion system of plant material in a liquid culture medium has the advantage over other in vitro culture systems, in that it provides a uniform distribution of nutrients, which results in a faster growth of plant material and an increase of the quality of the material obtained.
    [0010] Plantform ® or Rita ® reactor systems have the advantage of taking up little space, making them easy to place and store in in vitro culture chambers. However, these reactor systems have the drawback that the liquid culture medium remains in contact with the receptacle during cultivation, making it difficult to change the medium, as well as managing the reactor to comfortably access the plant material during the cutting phase. and collection.
    [0012] There are other reactor systems, such as the Setis ® brand reactor systems, which include two independent receptacles with their respective covers that communicate with each other. A first receptacle is used for the growth of the plant material while a second receptacle is used to store the liquid culture medium, which is periodically pumped through a connector to the first receptacle to carry out the temporary immersion of the plant material in the culture medium. The first receptacle in which the plant material is housed is stacked on the second receptacle and includes an inclined bottom to facilitate drainage of the liquid culture medium.
    [0014] The Setis ® reactor system has the advantage that it facilitates the change of the medium during the growth phase of the plant material, since it has a separate receptacle that can be disconnected to sterilize and replace the liquid culture medium that is housed in its inside. However, this reactor system has the drawback that it takes up space in the in vitro culture chambers. In addition, the presence of two receptacles makes it difficult to manage the system, and in particular, access to plant material for cutting and harvesting.
    [0016] Description of the Invention
    [0018] The objective of the present invention is to provide a reactor system for the in vitro cultivation of plant material that solves the aforementioned drawbacks, presenting the advantages that will be described below.
    [0020] In accordance with this objective, according to a first aspect, the present invention provides a reactor system for the in vitro cultivation of plant material, comprising a receptacle for the cultivation of plant material, a lid adapted to close an opening of said receptacle , and means to allow the entry and / or exit of gas from said receptacle.
    [0022] The system is characterized in that it comprises a container for receiving a liquid culture medium inside the receptacle and means for supporting the plant material on said reception container, where said container is attached to the lid of the receptacle in a position suitable for use, said lid including at least one medium inlet and / or outlet conduit that communicates with the container to be able to fill and / or empty said container through said conduit.
    [0024] In the claimed reactor system, the culture medium is pumped from the outside through a conduit that communicates the lid with the container that receives the liquid medium and in turn supports the plant material. This container is attached, preferably integrally, to the lid of the culture receptacle in a position suitable for use, so that when removing the lid, the container on which the vegetal material. Thanks to this, access to plant material for cutting or harvesting is very comfortable and easy.
    [0026] Another advantage of the claimed system lies in the fact that the culture medium never comes into contact with the culture receptacle, since the entry and exit of the medium occurs through at least one conduit that directly communicates the cover with the medium receiving container. Thanks to this, the dimensions of the culture receptacle can be reduced, and also, the shape and size of the receptacle can be very varied, with the only condition that the dimensions of the opening always make possible the introduction of the container attached to the lid. in a position suitable for use. In fact, in the claimed system, the receptacle may be, for example, a conventional receptacle of glass, or other material, which are commercially available for multiple uses, including, for example, an opening provided with a susceptible thread to be closed and sealed by a system cover adapted with a complementary thread.
    [0028] According to a second aspect, the present invention provides a kit for transforming a receptacle into a reactor for the in vitro cultivation of plant material, characterized in that it comprises;
    [0029] a) a lid adapted to close an opening of said receptacle, b) a container for receiving a liquid culture medium inside the receptacle, said container being attached to the lid of the receptacle in a position suitable for use, and including said lid at least one inlet and / or outlet conduit of the culture medium that communicates with the container to be able to fill and / or empty said container through said conduit, and c) means for supporting the plant material on said container for receiving the culture.
    [0031] The present invention provides a kit that allows a receptacle, for example a commercial receptacle of various shapes, dimensions and materials, to be transformed into a reactor for the in vitro cultivation of plant material suitable for the claimed system. This reactor can be used to cultivate in vitro plant material for industrial use, research use or domestic food use. For example, the kit can be used to transform a conventional receptacle into a reactor that allows in vitro cultivation of soybeans whose sprouts can have commercial or domestic food-grade use.
    [0033] In particular, the kit provides a lid adapted to close the opening of a receptacle. This cover may be, for example, a cover provided with a thread and adapted to close and threading sealing an opening of a conventional receptacle, for example, a conventional receptacle of glass, or other material, of those commercially available for multiple uses.
    [0035] The lid of the kit has the particularity that it is attached to a receiving container for a liquid culture medium on which the plant material is supported in a position suitable for use, so the opening of the receptacle must allow the passage of this container to attach the lid. By removing the same lid or receptacle, the receiving container that includes the culture medium and plant material is removed or accessed, leaving the culture receptacle free and intact. The reception container is filled through a duct provided in the lid itself to feed the culture medium from an external tank.
    [0037] According to one embodiment, the culture medium receiving container is attached to the lid so that in a fit-for-use position it extends longitudinally inside the culture receptacle from an inside face of the lid body. For example, the container can extend longitudinally inside the culture receptacle attached to the lid so that it does not contact the walls of this receptacle. Thanks to these characteristics, unlike the state-of-the-art reactor systems, the claimed reactor system allows working with smaller volumes of culture medium, saving costs and space.
    [0039] Preferably, according to a preferred embodiment, the body of the cap comprises a support base configured to be able to keep the receptacle and the container for receiving the liquid culture medium in a stable horizontal position, when the receptacle is attached to the cap.
    [0041] This particular configuration of the lid has the advantage that it allows to maximize the useful cultivation area of the container for receiving the culture medium, which extends longitudinally attached to the lid in a horizontal position inside the receptacle. Thanks to this it is possible to cultivate a large number of units of plant material in a very comfortable way. Another advantage lies in the fact that the technician can manipulate the plant material keeping the container and lid assembly in a stable horizontal position.
    [0043] Advantageously, the end of the receiving container attached to the interior face of the lid, includes increased side walls adapted to come into contact with the opening. of the receptacle. These side walls are attached to the opening of the receptacle and have the function of preventing the plant material from falling out of the receptacle, or avoiding the contact of the plant material with the external parts of the receptacle or with the technician's hands at the time of extraction. of plant material.
    [0045] Again advantageously, the culture medium receiving container includes a cavity provided with a hole to be able to actively or passively drain the liquid culture medium to the outlet and / or culture medium inlet duct of the lid. Optionally, the inner bottom of the receiving container can be configured with a slope to facilitate the drainage of the liquid to the exit hole. This hole can be used to inject through it the liquid culture medium from a conduit associated with an external tank in which said culture medium is stored.
    [0047] Preferably, the lid of the receptacle comprises at least one hole for the gas inlet and / or outlet of the receptacle. A gas can be injected through this hole to modify the interior atmosphere of the receptacle, if necessary.
    [0049] Again preferably, the plant material support means comprise at least one body, for example, a platform-like body, adapted to be removably coupled to the container receiving the liquid culture medium, said body being configured to allow the contact of the liquid culture medium with the plant material.
    [0051] According to an embodiment, the body on which the plant material is supported comprises a support platform provided with a plurality of perforations to allow the passage of an upward flow of liquid culture medium. This platform is adapted to be movably anchored inside the container, maintaining a predetermined distance from the bottom of the container itself.
    [0053] According to a different embodiment, instead of a platform, the body that supports the plant material and allows the passage of liquid can be configured, for example, as a fabric, such as, for example, a porous tissue, or a permeable membrane, or inert material that allows the passage of liquid. This body will be arranged on the receiving container of the culture medium.
    [0055] Advantageously, the system comprises means for holding the plant material capable of growing on the receiving container of the culture medium. These holding means are they are removably coupled on the receiving container, or more specifically, on the support platform that is anchored to the receiving container of the culture medium so as to allow the passage of shoots or growing parts of the plant material.
    [0057] Preferably, the support means are configured as a grid, so that the framework of this grid supports the base of the plant material that grows on the receiving container or, more specifically, on the platform for supporting said plant material.
    [0059] Holding the plant material by means of a grid, or any other similar or equivalent means, facilitates the cutting and harvesting of shoots, since it maintains the explants in position, without the need to manipulate them manually, when the receiving container tilts or inverts subject by the cover to facilitate the cutting and collection operations of the plant material. When these operations are carried out, the cavity of the container is free of liquid culture medium, since the medium has previously been drained through the liquid outlet duct of the lid.
    [0061] Again preferably, the claimed reactor system comprises means for pumping with a predetermined frequency the liquid culture medium through the at least one culture medium inlet and / or outlet conduit of the lid, said pumping enabling temporary immersion of the plant material arranged on the support means of the receiving container of the culture medium.
    [0063] In accordance with a third aspect, the present invention provides a Method for cultivating plant material in vitro in a receptacle by the claimed reactor system, comprising the steps of;
    [0064] a) injecting a liquid culture medium into the culture receptacle through at least one inlet duct provided in a lid attached to a culture medium receiving container,
    [0065] b) temporarily immersed in the liquid culture medium a plant material previously deposited on the receiving container that is attached to the lid,
    [0066] c) actively or passively drain the liquid culture medium through an outlet duct provided in the lid,
    [0067] d) after step c) access the container for receiving the culture medium and all the plant material deposited on the container, to carry out the cutting and collection on the outside of the receptacle.
    [0068] The claimed in vitro cultivation method has the advantage that it allows obtaining a higher multiplication rate of the plant material, since the shoots are longer, have a higher fresh weight and with larger leaves than those obtained with other cultivation methods. This is so thanks to the fact that the reactor system allows to practice an optimal temporary immersion frequency of liquid medium, guaranteeing at all times also an optimal drainage of the medium through the outlet duct provided in the lid and that communicates directly with the drain hole of the container itself.
    [0070] On the other hand, the claimed method allows access to the plant material in a very easy way, removing the lid or the receptacle to access the receiving container and the plant material that is supported on the receiving container. Once outside the receptacle, the container can be inverted or tilted to facilitate cutting and harvesting of plant material. Furthermore, for the embodiments in which the lid is provided with a support base configured to be able to keep the container itself in a stable horizontal position, the handling of the plant material is extremely agile.
    [0072] Optionally, the claimed method comprises the step of modifying the interior atmosphere of the culture receptacle by injecting gas through at least one conduit of the lid that communicates with the interior of the receptacle.
    [0074] In the present invention;
    [0076] Plant material means any type of material capable of growing, or of being vegetatively multiplied or propagated, for example, a material that contains or consists of plant tissues, plant organs, seeds and / or plant cells, preferably, said material may consist in explants or living tissues isolated from its own organ, prepared aseptically.
    [0078] A culture receptacle means a container or container in which the growth of the plant material is carried out.
    [0080] In vitro cultivation means a cultivation of plant material for industrial use, research use, commercial food use or domestic use, capable of being carried out using the claimed system either in industrial facilities, research laboratories, commercial premises, or in a user's home. In vitro culture comprises plant material, such as explants or parts of a plant or seed, which is carried out under controlled temperature, photoperiod and light intensity conditions, with culture media formulated according to predetermined objectives. Optionally, the plant material may comprise biotic agents, for example bacteria and / or fungi, or abiotic agents, for example chemical compounds added to the liquid culture medium.
    [0081] Brief description of the figures
    [0083] For a better understanding of what has been stated, some drawings are attached in which, schematically and only by way of non-limiting example, a practical case of embodiment is represented.
    [0085] FIG. 1 is a perspective view of one embodiment of the culture receptacle of the reactor system of the present invention.
    [0087] Figure 2 is a perspective view of a means of holding the plant material configured as a grid that forms a lattice.
    [0089] Figure 3 is a perspective view of a means for supporting the plant material on the culture medium receiving container, which are configured as a platform provided with a plurality of perforations to allow the upward flow of the liquid culture medium.
    [0091] Figure 4 is a perspective view of a lid and liquid culture medium receiving container assembly, which is attached to the lid on its inner face. This lid is adapted to close the opening of the receptacle of figure 1.
    [0093] Figure 5 is an exploded perspective view schematically showing the assembly of the reactor system with the claimed kit.
    [0095] Figure 6 is a perspective view showing the reactor system in the cultivation phase, when the culture medium receiving container is fed from the outside through a conduit provided in the lid to temporarily immerse the plant material . The arrows indicate the entrance of the liquid medium that causes the simultaneous exit of air from the interior of the culture receptacle.
    [0097] Figure 7 is a perspective view showing the reactor system in the cultivation phase, when the liquid culture medium leaves the container through the conduit of the lid. The arrows indicate the exit of the liquid medium that causes the simultaneous entry of air into the culture receptacle.
    [0099] Description of a preferred embodiment
    [0101] A preferred embodiment of the system and kit of the present invention is described below with reference to Figures 1 to 7.
    [0102] As discussed in the previous sections, the present invention provides a reactor system and a kit suitable for transforming a conventional receptacle 1, such as that shown in Figure 1, into a reactor for the in vitro cultivation of plant material.
    [0103] In the described embodiment, the kit consists of the following components;
    [0105] - a cover 2 to close the receptacle 1 where the cultivation takes place,
    [0106] - a container 4 for receiving the liquid culture medium (not shown) which is attached to the lid 2,
    [0107] - a body 6 as a platform with perforations 6a that allows to support the plant material 11 on the container 4,
    [0108] - a grid-like body 7 capable of being removably coupled on the body 6 or support platform, in order to hold the base of the plant material 11 and facilitate the cutting and harvesting of the shoots of said plant material 11, - a conduit 5 for the entry and / or exit of the liquid culture medium through the cover 2, and
    [0109] - two holes 8, 9 for the entry and / or exit of gases from the culture receptacle 1.
    [0111] The lid 2 can be, for example, a lid 2 provided with a thread 2a and adapted to close and thread seal 1 the opening 3 of the conventional receptacle 1, which in the described embodiment is a receptacle 1 of glass or polycarbonate , commercially available for multiple uses. Attached to the cover 2 is a sealing gasket 10 to ensure the sealing of the assembly.
    [0113] The reactor system includes means (not shown) for pumping, at a predetermined frequency, the liquid culture medium from an external tank (not shown) to the receiving vessel 4.
    [0115] In the described embodiment, the cover 2 has the particularity that it includes a support base 2b configured to be able to keep the receptacle 1 and the container 4 for receiving the culture medium in a stable horizontal position. As can be seen in Figures 4 to 7, the container 4 for receiving the culture medium extends longitudinally joined integrally to the lid 2, from an inside face of said lid 2, so that it is arranged in a horizontal position stable, substantially parallel to the walls of the culture receptacle 1a, but without contacting these walls.
    [0117] As discussed in the description of the invention, this particular configuration allows to maximize the useful cultivation area and also to work with smaller volumes of culture medium, which saves cost and space.
    [0118] Figure 4 shows a perspective view of the assembly of lid 2 and receiving container 4 attached to lid 2. This figure shows the inclined bottom 4b for the active or passive drainage of the culture medium through a hole that communicates with the cover 2 and the outlet duct 5 of liquid culture medium. The side walls of the container 4 include projections 4a that serve as a support base for the body 6 or platform on which the plant material is supported. These projections 4a are positioned halfway up the walls of the container 4, so that the body 6 or platform on which the explants are arranged can be flooded with liquid medium when the culture medium passes through the perforations 6a (see figure 3).
    [0120] Continuing with the description of figure 4, it is observed that the container 4 includes at the end that contacts the inner face of the lid 2, increased walls 4d adapted to fit into the walls of the opening 3, so as to prevent spillage of liquid medium when the technician slightly tilts the reactor or the assembly of cover 2 and receiving container 4 to, for example, carry out the cutting of the plant material.
    [0122] Figure 2 shows a perspective view of the body 7 as a grid, provided with a framework that is crossed by the sprouts of the plant material 11 (see figures 6 and 7). This lattice serves to hold the base of the explants when container 4 is tilted to facilitate rapid cutting and harvesting of shoots. This grid or body 7 includes lateral walls 7a that are arranged on the support platform or body 6, at a certain distance from said platform.
    [0124] As mentioned in the description of the invention, unlike the state-of-the-art reactor systems, the reactor system of the present invention has the advantage that the collection or cutting of the shoots is carried out by accessing the lid 2 and container 4 assembly from the outside of the receptacle 1, so that the technician can even slightly tilt this assembly to facilitate the rapid cutting of the plant material 11 in different positions.
    [0126] The method of in vitro cultivation of plant material 11 by means of the claimed reactor system described in the attached figures is described below.
    [0128] Once the different components have been assembled following the arrangement shown schematically in Figure 5, air filters (not shown) are connected to the gas inlet and outlet ports 8 and 9 of cover 2, and a tube to the inlet and outlet conduit 5 of liquid culture medium. The set is sterilized before being used for the cultivation of the explants in order to guarantee sterile conditions.
    [0129] After sterilization, the container 4 and lid 2 are accessed inside a laminar flow cabinet and, in sterile conditions, the plant material 11 is deposited on the platform or body 6, taking advantage of the openings left by the body's framework 7th grating of the explants.
    [0131] Once the plant material 11 has been deposited, the container 4 and cover 2 assembly is introduced inside the culture receptacle 1 and it is placed in an in vitro culture chamber, under controlled conditions of temperature, photoperiod and light intensity. The liquid culture medium is then injected into the receptacle 1 through the inlet duct 5 provided in the cover 2 until the plant material 11 is immersed in the medium. The injection is carried out periodically from an external tank (not shown) by means of pumping, and using timers.
    [0133] During the cultivation, the plant material 11 is temporarily immersed according to the temporary immersion cultivation system, already known in the state of the art. The immersion phase is short, of a few minutes, to moisten the explants, leaving them impregnated with culture medium. When the injection ceases, the medium is drained from the inclined bottom 4b through the same conduit 5 of the cover 2, which at this time acts as an outlet conduit (see figure 7). During the injection, the air inside the receptacle 1 exits through the holes 8, 9 provided in the cover 2 and associated with the filters (not shown) (see figure 6). However, by stopping the injection and draining the medium, fresh air enters through the same ports 8 and 9. If necessary, the interior atmosphere of the culture receptacle 1 can be modified by injecting a gas.
    [0135] The plant material 11 remains impregnated with medium for hours, growing inside the culture receptacle 1 under controlled light and temperature conditions. When cutting or harvesting is necessary, the technician accesses the container 4 and lid 2 assembly, in this case unscrewing the receptacle 1. The lid 2 support base 2b allows the container 4 to be kept in a stable horizontal position to carry out the cut accurately.
    [0137] As mentioned in the description of the invention, the cultivation method described allows obtaining a higher multiplication rate of the plant material, 11 the shoots being longer, with a higher fresh weight and larger leaves, than those obtained with other cultivation methods that employ other reactor systems.
    [0139] The table below shows the results of a culture test carried out using a Rita ® brand reactor system and the claimed reactor system.
    [0142] The results show that the number of initial explants per culture vessel is much higher in the reactor system of the present invention. The number of final shoots obtained and the average length and weight of the shoots are also higher.
    [0144] Although reference has been made to a specific embodiment of the invention, it is evident to a person skilled in the art that the described reactor system and kit is susceptible of numerous variations and modifications, and that all the mentioned details can be replaced by others technically equivalent, without departing from the scope of protection defined by the appended claims. For example, although an embodiment has been described in which the cover 2 includes a support base 2b and the receptacle 1 is of a conventional type, the same reactor system would be viable with a receptacle that includes a support base that would also allow maintaining the assembly in a stable horizontal position. In this case, it would not be necessary for the lid to have a support base and the lid and container assembly could still be removed for cutting and harvesting, even though, once removed from the receptacle, it alone could not be kept in a stable horizontal position. Similarly, although plant support and support means 11 configured as a platform and grid have been described, similar results could be obtained with other plant material support and support means provided that said means allowed the material The vegetable could also be impregnated by the culture medium stored in the cavity of the container 4 attached to the lid 2.
    权利要求:
    Claims (22)
    [1]
    1. Reactor system for the in vitro cultivation of plant material, comprising a receptacle (1) for the cultivation of plant material (11), a cover (2) to close an opening (3) of said receptacle (1) , and means (8, 9) to allow the entry and / or exit of gas from said receptacle (1), characterized in that it comprises a container (4) to receive a liquid culture medium inside the receptacle (1 ), and means (6, 7) for supporting the plant material (11) on said receiving container (4), where said container (4) is attached to the lid (2) of the receptacle (1) in a position suitable for use, said cover (2) including at least one inlet and / or outlet duct (5) of medium that communicates with the container (4) to be able to fill and / or empty said container (4) through said duct (5) ).
    [2]
    The reactor system according to claim 1, wherein the culture medium receiving container (4) is attached to the lid (2) so that, in a fit-for-use position, it extends longitudinally from one side of the body of the cover (2) inside the receptacle (1).
    [3]
    3. Reactor system according to any of claims 1 to 2, wherein the body of the lid (2) comprises a support base (2b) configured to be able to maintain the receptacle (1) and the receiving container (4) of the liquid culture medium in a stable horizontal position.
    [4]
    Reactor system according to any of claims 2 to 3, in which the end of said receiving container (4), which is attached to the inner face of the lid (2), includes walls (4c, 4d) regrowth sides adapted to come into contact with the opening (3) of the receptacle (1) and facilitate cutting of the plant material without damaging the cover (2).
    [5]
    5. Reactor system according to claim 1, wherein said culture medium receiving container (4) includes a cavity provided with an inclined bottom (4b) and a hole for draining the liquid culture medium to the at least conduit (5) exit and / or entry of culture medium.
    [6]
    6. Reactor system according to any of claims 1 to 5, wherein said support means for the plant material (11) comprise at least one body (6) adapted to be removably coupled to the container receiving the culture medium liquid, said body being configured to allow contact of the liquid culture medium with the plant material (11).
    [7]
    7. Reactor system according to any of claims 1 to 6, comprising means (7) for holding the plant material (11) capable of growing on said support means coupled to the container (4) for receiving the culture medium.
    [8]
    Reactor system according to claim 7, wherein said holding means (7) are configured as a grid, the framework of said grid being capable of holding the base of the plant material (11) growing on the container ( 4) receiving the culture medium.
    [9]
    9. System according to claim 1, wherein said gas inlet and / or outlet means of the receptacle comprise at least one hole (8, 9) provided in the cover (2) for the inlet and / or outlet of gas.
    [10]
    10. Reactor system according to any of the preceding claims, wherein said receptacle is provided with an opening (3) dimensioned to allow the introduction of the container (4) for receiving the liquid culture medium that is attached to the lid (2 ) closing the receptacle (1).
    [11]
    11. Reactor system according to any of claims 3 to 10, wherein the opening (3) of said receptacle includes a thread (1a) and said cover (2) is adapted to screw-close and seal said opening (3) , said cover (2) comprising a support base (2b) configured to keep the receptacle (1) and the container (4) in a stable horizontal position while said receptacle (1) is coupled to the cover (2).
    [12]
    12. System according to any of the preceding claims, comprising means for pumping with a predetermined frequency the liquid culture medium through said at least one conduit (5) for entry and / or exit of culture medium from the cover (2 ), said pumping making possible the temporary immersion of the plant material (11) arranged on the support means (6) of the container (4) for receiving the culture medium.
    [13]
    13. Kit for transforming a receptacle (1) into a reactor suitable for the in vitro cultivation of plant material (11), characterized in that it comprises;
    a) a lid (2) adapted to close an opening (3) of said receptacle (1), b) a container (4) to receive a liquid culture medium inside the receptacle (1), said container being attached to the lid (2) of the receptacle (1) in a position suitable for use, and said lid (2) including at least one inlet and / or outlet duct (5) of the culture medium that communicates with the container (4) for to be able to fill and / or empty said container (4) through said conduit (5), and
    c) means (6) for supporting the plant material (11) on the container (4) for receiving the culture medium.
    [14]
    Kit according to claim 13, in which the culture medium receiving container (4) is attached to the lid (2) so that it extends longitudinally from one face of the lid body (2).
    [15]
    Kit according to any of claims 13 to 14, in which the cover (2) includes a support base (2b) adapted to be able to keep the container (4) for receiving the culture medium in a stable horizontal position.
    [16]
    Kit according to any one of claims 13 to 15, in which said means for supporting the plant material (11) comprise at least one body (6) capable of being removably coupled to the container (4) for receiving the liquid culture, said body (6) being configured to allow contact of the liquid culture medium with the plant material (11).
    [17]
    17. Kit according to any of claims 13 to 16, comprising means (7) for holding the plant material (11) when said material grows on the container (4) for receiving the culture medium.
    [18]
    Kit according to claim 17, in which said fastening means (7) are configured as a grid, the framework of said grid being susceptible holding the base of the plant material (11) capable of growing on the support body (6) coupled to the container (4) for receiving the culture medium.
    [19]
    Kit according to any one of claims 13 to 18, wherein said cover (2) is adapted to screw close and seal a threaded opening (3) of a commercially available receptacle.
    [20]
    20. Method for cultivating plant material in vitro by means of the reactor system according to any of claims 1 to 12, comprising the steps of;
    a) injecting a liquid culture medium into the receptacle (1) through at least one inlet conduit (5) provided in a lid (2) attached to a container (4) for receiving the culture medium,
    b) temporarily keep immersed in the liquid culture medium a (11) plant material previously deposited on the receiving container (4) that is attached to the lid (2),
    c) actively or passively drain the liquid culture medium through an outlet duct (5) provided in the cover (2),
    d) after step c) access the receiving container (4) attached to the lid (2) to cut and collect from the outside of the receptacle (1).
    [21]
    21. Method according to claim 20, comprising the step of modifying the interior atmosphere of the receptacle (1) by injecting gas through at least one duct (8, 9) of the cover (2) that communicates with the interior of the receptacle ( one).
    [22]
    22. The method according to any of claims 20 to 21, wherein the cover (2) includes a support base (2b) adapted to be able to keep the receptacle and the container (4) for receiving the culture medium in a horizontal position stable while said receptacle (1) remains attached to the cover (2).
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    同族专利:
    公开号 | 公开日
    CL2021001417A1|2021-11-12|
    ES2763637B2|2020-10-14|
    WO2020109637A1|2020-06-04|
    US20220022396A1|2022-01-27|
    EP3888453A1|2021-10-06|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO2012044239A1|2010-09-30|2012-04-05|Plant Form Ab|Bioreactor for in vitro culture of plants|
    WO2012156440A1|2011-05-18|2012-11-22|Sopet Nv|Container system for immersion growth regime|
    WO2014072962A1|2012-11-12|2014-05-15|Universidad Católica Del Maule|A temporary immersion bioreactor method and relative product|
    GB2533129A|2014-12-11|2016-06-15|Alkion Biopharma Sas|Bioreactor|
    MX2016015393A|2016-11-24|2018-05-23|Inst Politecnico Nacional|System for the temporary immersion of a single growth chamber for in vitro culture of plants.|
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