Container and method for in-vitro plant cultivation

Abstract

A lower part (2) of a container (1) closed media-tightly by an upper part (3) has a receiving chamber (8) for a culture medium, said receiving chamber being divided by a grating (6) into a plurality of cells (7). For improved handling when transplanting, a separate grating (6) is provided which has cell bases (18) which are flexible and / or can be pushed through and / or are at least partially removable. The bottom (4) of the lower part can also be divided by the adjacent grating into cell bases which are flexible and / or can be pushed through and / or are at least partially removable, so that the plants (15) can be transplanted without having to remove the grating (6). According to a second concept of the invention, at least one portion of a side wall (5) of the lower part (2), which side wall is taller than the grating (6), is reduced and / or can be reduced in height and / or can be opened.

Description

[0001] Containers and methods for in vitro plant cultivation

[0002] The present invention relates to a container for in-vitro plant cultivation according to the preamble of claim 1, as well as a method for in-vitro plant cultivation.

[0003] In vitro plant cultivation involves cloning plant explants on a culture medium in a sterile environment into complete and genetically identical plants. Transparent plastic containers are used. These consist of a cup-shaped lower section and a lid attached to the upper section in a media-tight manner. Grid-shaped inserts are used to facilitate the orderly formation of the meristems. These form individual cells within the receiving space, from which the newly formed plants can be selectively removed. The grid can be removed from the lower section, particularly for automated transplanting. A container of this type equipped with a grid is known from JP S63287478 A.

[0004] The object of the present invention is to provide a container for in vitro plant cultivation that improves handling of the container, particularly during automatic plant transplanting. A further object is to provide a related method.

[0005] The object is achieved by a container according to claim 1 and a container according to claim 8. The method is specified in claim 12. Advantageous embodiments of the invention can be found in the dependent claims referring back to these claims as well as in the following description of advantages and figures.

[0006] According to a first inventive concept, it is provided that the bottom of the lower part is divided into respective cell bottoms by the adjacent grid and / or the grid has its own bottom with respective cell bottoms, wherein the respective cell bottoms are flexible and / or pierceable in the direction of the cell interior and / or the bottom is at least partially removable from the lower part or the grid.

[0007] The flexibility and / or penetrability of the cell bottoms and / or the ability to pull the bottom off the base or the grid make it possible to lift the newly formed plantlets on the nutrient medium out of the grid using pestles positioned underneath and to push them at least partially upwards so that they can be more easily grasped and transplanted into the next rearing and / or cultivation container, e.g. a rearing tray. Preferably, the cells contain a nutrient medium that solidifies into (agar) plugs, e.g. gelling agar-agar, or porous plugs made of e.g. rock wool or foamed material that absorb the nutrient medium, so that the plantlets are lifted together with the plugs on which they have grown and are at least partially pushed out of the grid. The plantlets can be more easily grasped using the plugs and, in particular, can be transplanted together with the plugs onto a subsequent substrate.This can then be done in particular automatically with transplanting machines, which remove the plugs with the plants from the grid, preferably row by row. The bottom of the lower section or the grid can remain closed until immediately before transplanting, thus avoiding contamination from any falling or adhering nutrient medium. This significantly improves and simplifies handling of the container and ensures a clean working environment. If the flexibility and / or pierceability of the cell bases and / or the removal of the soil from the lower section of the container is realized, the previously necessary removal of the grid from the lower section can be eliminated. Transplanting can be done directly from the container or lower section in a way that is gentle on the plants. The cup-shaped lower section with the grid contained therein is also easier to handle, especially when automated, than a grid removed from the lower section.On the other hand, a grid that can be removed from the base can be handled better and, in particular, automatically if the grid is closed with a flexible and / or pierceable and / or removable base.

[0008] The grid extends upwards from the bottom, with the grid walls preferably tapering towards the top. The plugs can be more easily lifted and removed from the conically widened cells. The grid can be a separate grid insert that can be arranged in the lower part, preferably directly adjacent to the bottom of the lower part, and preferably secured in the lower part, e.g., by means of a positive connection such as a snap connection. The cell size and the number of cells in the container depend in particular on the plant species and are particularly tailored to the number of trays in the subsequent cultivation tray. The preferably square-shaped cells have a width of 5 to 50 mm and a height of 5 to 50 mm, enclosing a cell volume of 0.125 to 125 ml. A common cell size has the dimensions W x W x H = 12.7 x 12.7 x 12.7 mm. 3 and encloses a cell volume of approximately 2 ml.

[0009] Flexibility is to be interpreted such that at least a section of the respective cell base can be deformed relative to the adjacent grid in the expulsion direction by a few millimeters, preferably by more than 5 mm, into the cell interior, while the remaining lower part or grid remains essentially dimensionally and form-stable. The same applies to penetrability, whereby the respective cell bases can be pierced comparatively easily and, above all, in a targeted manner and limited to the respective cell base by pestles, preferably with cutting edges. The pestles, which preferably move continuously, can also expel the plants from the cells immediately after penetration.

[0010] The flexibility and / or puncturing ability and / or peeling ability can be provided by a film- or foil-like base with a thickness of 45 to 200 μm. The preferably multi-layered film can contain a foamed, microperforated and / or membrane layer for this purpose. Due to this flexibility, the base is particularly reversibly deformable, so that the container can also be used multiple times in in vitro plant cultivation. The flexibility and / or puncturing ability of the base is matched to the lattice structure, so that the respective cell bases deform essentially point-symmetrically or tear open in the middle when pierced. In particular, the base can have a thinner wall thickness within the cells than in areas directly adjacent to the lattice.Sufficient flexibility or easy penetrability of the cell bottoms can also be achieved by making the bottom of a different, in particular more elastic or more brittle, plastic than the other lower part, whereby the lower part can be manufactured by 2K injection molding.

[0011] The container, which consists of the lower and upper parts as well as the grid, is therefore preferably made entirely of a preferably identical transparent plastic, such as polypropylene (PP) or polystyrene (PS), so that plant growth can be visually monitored, particularly through the ground. The lower part preferably has a flange- and / or apron-shaped peripheral edge, which can be arranged at the end opposite the ground or laterally on the side wall and with which a peripheral edge on the upper part cooperates to seal the container in a media-tight manner. A gas-permeable membrane can be inserted in the upper part, preferably in its lid. In a preferred further embodiment, the grid is connected integrally to the lower part.The grid is therefore not a separate component, but an integral part of the base, ensuring it is securely integrated and, above all, securely positioned. When the nutrient medium is added, the grid does not float and does not require special support during transport. Preferably, the base of the base is at least partially connected to the grid to form closed cells at the bottom, preventing the plants from rooting together and allowing the plugs containing the newly formed plants to be gently removed and replanted from the base of the container.

[0012] In a preferred further embodiment, the base is formed by a label. The label, punched from a thin film, can be glued or sealed to the lower part or grid produced with an open base by injection molding or thermoforming, or can be integrated into the further lower part or grid as an in-mold label (IML) during injection molding. The film can be a multilayer film and contain a foamed, microperforated, and / or membrane layer.

[0013] Preferably, the label has at least one grip tab, which allows the label to be easily removed at least partially from the base or the grid. The plugs can be easily pushed out of the exposed cells on the underside of the grid.

[0014] In a preferred further embodiment, the respective cell bases of the lower part or the grid in particular have (pre-)embossed thin spots. The cell bases tear open at the thin spots when pierced. This can also enable the cell bases to be punched out. Additionally or alternatively, hinges can be formed in the cell bases through thin spots, through which the pushed-open cell bases remain connected to the further lower part or grid and are not removed together with the plugs. The thin spots can be formed by appropriately designed tool molds and / or formed by embossing carried out subsequent to or integrated with the injection molding and / or pre-embossed in a label forming the base.

[0015] In a preferred further embodiment, the cell bottoms are convex and / or concave when viewed from below. The basic shape of the cell bottoms is in particular convex, so that correspondingly rounded plugs are present, which can be more easily inserted into a subsequent substrate. The convex basic shape of the flexible cell bottoms can be changed into a concave shape by means of a tappet, whereby a first lifting and thus release of the plugs from the cells takes place without the cell bottoms tearing open. The cell bottoms can then be pierced for a second further lifting. Alternatively, if the plugs can already be removed with the first lifting, the cell bottoms can be pressed back into their convex basic shape without causing any damage. A small central section of the cell bottom can be concave, concentric to the convex basic shape, so that a receiving section is provided which centers the tappet.

[0016] In a preferred further embodiment, the walls of the grid surrounding the cells are provided with recesses. These recesses, particularly in the upper region of the cell walls, form overflow channels, allowing agar added to one or a few places to be evenly distributed over the cells. The (agar) plugs, which subsequently solidify, are connected only by the narrow webs of the overflow channels, which are easily torn off when the plugs are removed.

[0017] According to a second inventive concept, the side wall of the lower part is raised relative to the grid, with at least one section of the side wall being reduced in height and / or reducible and / or openable. The side wall, which is raised relative to the grid, creates a protected space for the newly forming plants. In particular, the raised side wall provides lateral support for the still weak plants. The side wall, which is reduced in height and / or reducible and / or openable, is preferably openable or can be opened on one side of the lower part up to the height of the grid, so that lateral access is provided to the plants or plugs, which have preferably been at least partially pushed out of the grid, while the plants continue to be supported by the further side wall. In particular, transplanting machines can thus transplant the plants or plugs more easily and automatically.Plugs can be grasped and removed from the base without first having to remove the trellis. The height-reduced and / or reducible and / or openable side panel also facilitates the removal of a separate trellis, preferably one with its own base, as the trellis can be grasped directly from the side without having to reach between the plants from above.

[0018] The height reduction of the side wall is relative to a plane parallel to the bottom of the lower part, in which the uppermost edge of the side wall lies. The upper edge of the side wall may be inclined relative to this plane and / or locally offset downward, particularly after the side wall has been reduced in height. To bridge openings and reductions preformed in the side wall of the lower part, the upper part has a correspondingly high side wall.

[0019] In a preferred further embodiment, it is provided that the at least one section of the side wall can be at least partially separated from the side wall. The at least one section of the side wall can be completely separated and thus removed from the lower part. The dividing lines can be defined by thin spots. An articulated connection to the further lower part can remain, so that the relevant side wall section can be folded out like a door or flap. In a preferred further embodiment, it is provided that the at least one section of the side wall is formed by an upper, circumferential side wall part which is removably attached to a lower side wall part. The upper side wall part, which is connected in particular in a gas-tight manner to the lower side wall part, can be separated from the lower side wall part and removed by a removable separating strip.The dividing line can be formed by a groove-like depression that is easily severed and, in particular, provides a guide for a cutting blade. The upper side wall section can be integrally connected to the lower side wall section via a shoulder, enclosing it in an apron-like manner. The upper side wall section can thus be punched out in the area of ​​the shoulder opposite the lower side wall section or, if predefined tear lines are present, can be easily pressed down. The respective dividing line or shoulder is preferably located approximately at the level of the grid. The upper side wall section can be a separate component that can be removably and media-tightly attached to the side wall of the lower section.

[0020] In a preferred further embodiment, a flange- and / or skirt-shaped peripheral edge of the lower part lies in an edge plane that runs at an acute angle to the standing plane of the container. The standing plane is defined by the bottom of the lower part or by at least one support web. Preferably, a complementarily designed upper part has an edge plane that runs at an acute angle to the lid plane, so that the container composed of the lower and upper parts in turn forms a support surface parallel to the standing plane for another container to be stacked on top.

[0021] In a preferred further embodiment, the bottom of the lower part is formed by a removable sealing film. From the cells, which can thus be opened at the bottom, the plugs can be lifted out of the grid by pushers inserted from below and at least partially pushed out, making them easier to grasp from the side. The method according to the invention comprises the following steps:

[0022] - Providing a container according to the first and / or second inventive concept;

[0023] - Introducing a nutrient medium into the cells of the receiving space of the lower part divided by the grid;

[0024] - Arranging sterile plant cell material on the nutrient medium;

[0025] - Close the container in a sterile manner;

[0026] - Let the plants grow;

[0027] - Opening the container;

[0028] - At least partial pushing of the plants upwards out of the cells of the grid;

[0029] - grasping the pushed-out plants; and

[0030] - Transplanting the plants into a second container.

[0031] In a preferred further embodiment of the method, the introduced nutrient medium solidifies into (agar) plugs and / or is absorbed by porous plugs inserted into the cells and that the plants are pushed out and grasped via the plugs.

[0032] In a preferred further embodiment of the method, it is provided that during or after opening the container, at least a section of the side wall of the lower part is reduced and / or opened.

[0033] In a preferred further embodiment of the method, after opening the container, a separate grid provided with its own base is removed from the container so that the plants outside the container are pushed out of the grid upwards.

[0034] In a preferred further embodiment of the method, the plants or plugs are pushed out of the cells at least partially upwards by means of pushers acting on the bottom of the lower part or the grid removed from the container, in that the pushers deform and / or pierce the respective cell bottoms due to their flexibility or the soil has previously been at least partially removed from the lower part or the grid.

[0035] Further advantages and details of the invention can be found in the following description of the figures. Individual technical features of the exemplary embodiments described below can also be combined with previously described exemplary embodiments as well as the features of the independent claims and any further claims to form subject matter according to the invention. Where appropriate, functionally equivalent elements are provided with identical reference numerals.

[0036] Fig. 1 a lower part provided with a grid;

[0037] Fig. 2 shows a section of the bottom of the lower part in a first embodiment;

[0038] Fig. 3 the bottom of the lower part in a second embodiment;

[0039] Fig. 4 a grid with its own base;

[0040] Fig. 5 - 7 sectional views of the lower part with different base designs;

[0041] Fig. 8 - 14 different embodiments of the container.

[0042] A grid 6 is integrated into the lower part 2 shown in Fig. 1, which divides the receiving space 8 delimited by a base 4 and a side wall 5 into individual cells 7. The grid 6, which is connected in one piece to the lower part 2, has recesses 10 in its walls as overflow channels, through which thin liquid agar introduced into the receiving space 8 as a nutrient medium can be evenly distributed over the cells 7 before it solidifies into (agar) plugs 16. The fill level is at the level of the upper edges of the grid 6 or slightly below, so that the plugs 16 are connected to one another at most by the narrow webs in the recesses 10, which are easily torn off when the plugs 16 are removed. The lower part 2 has a flange-shaped peripheral edge 9, to which an upper part 3 can be fastened in a media-tight manner.

[0043] 2 and 3 show first and second embodiments of the lower part 2, in which the base 4 or the individual cell bases delimited by the grid 6 can be pierced by deliberately formed thin spots 13, 14, as shown in Fig. 5. A plunger 17 is pressed from below against the respective cell base 18, which tears open at the thin spots 13, 14 when a certain pressure force is exceeded. The plunger 17 can penetrate further into the respective cell 7 and push the plug 16 with the plantlet 15 formed on it out of the grid 6 far enough that a gripper (not shown in detail) of a transplanting machine can grasp the plug 16 laterally and pull it out of the grid 6.

[0044] Fig. 4 shows a separate grid 6, which can be arranged in a container and removed from it again at the end of a cultivation phase. The grid 6 is sealed on the underside with a label 11 forming its own base 11. The label 11 is attached as a sealing film to the lower edges of the grid 6. It can be removed from the additional grid 6 via a laterally protruding grip tab 12. The cells 7 are then opened at the bottom, and the pusher 17 can push the plug 16 with the plantlet 15 formed on it out of the grid 6, as shown in Fig. 6.

[0045] A further embodiment of a lower part 2 can be seen in Fig. 7. It shows a cross-section of convex-shaped cell bottoms 19, wherein a central section is concavely shaped concentrically to the convex basic shape as a centering receiving section for the plunger 17 pressing against it. The cell bottoms 19 are flexible, so that the plunger 17 can reshape the respective cell bottom 19 into a concave shape without destroying the cell bottom 19. The plugs 16 are thereby demolded from the respective cells 7 and lifted relative to the grid 6. The lower part 2 stands on support webs 20. The embodiments of the container 1 consisting of a lower part 2 and an upper part 3 shown in Figs. 8 - 14 can in particular be combined with the base designs of the lower part and grid described above. In Fig. 8, an overall flat lower part 2 is shown, as in Fig.1, combined with an upper part 3, which has a high side wall 21 compared to the lower part, which extends downward from a lid 22 provided with filter membranes 24 and encloses a correspondingly large space for the developing seedlings 15. With an edge 23 opposite the lid 22, the upper part 3 can be secured in a media-tight manner to the edge 9 of the lower part 2. To transplant the seedlings 15, the upper part 3 is removed from the lower part 2.

[0046] Fig. 9 shows a lower part 2 in which the side wall 5 extends upwards beyond the circumferential edge 9 on three sides, so that the seedlings 15 are still supported laterally even when the upper part 3 is removed. The fourth side, which is not raised or reduced in height, forms a lateral opening 25 through which a gripper of a transplanting machine can move into the lower part 2 and directly grasp and remove the grid 6 or the seedlings. In the lower part 2 shown in Fig. 10, such an opening 25 is created by a wall section 26 being removable from a high, circumferential side wall 5. The corresponding upper part 3 has a significantly lower side wall 21.

[0047] Fig. 11a shows a lower part 2 in which the circumferentially high side wall is divided into a lower side wall part 27 and an upper side wall part 28. The two side wall parts 27, 28 are connected to one another in a media-tight manner via a shoulder 32, wherein the shoulder 32 has a tear line at which the connection tears open when the upper side wall part 28 is pressed down relative to the lower side wall part 27. The upper side wall part 28 separated from the lower part 2 can be removed or, as shown in Fig. 11b, can accommodate the lower side wall part 27. Figs. 12 and 13 show a lower part in which an edge 29 is formed which runs at an acute angle to the standing plane. The corresponding upper part 3 has a complementary edge 31, so that the container 1, composed of the lower and upper parts, in turn forms a support surface parallel to the standing plane for another container to be stacked on top. In the lower part 2 in Fig.12, the side wall 5 is shaped to correspond to the inclined edge 29, so that at least on one side of the lower part there is unhindered access to the grid and the plants located in its cells. The adjoining sides of the side wall 5 rise continuously up to the opposite, raised side and offer at least partial protection for the plants located in the lower part. In particular, the plants located in the rear rows of the grid, which are removed last, are sufficiently supported by the correspondingly raised areas of the side wall. In the embodiment shown in Fig. 13, the plants in the front rows are also well supported laterally, namely in which one side wall 30 is raised relative to the edge 29 in the area of ​​the inclined edge sections.

[0048] Fig. 14 shows a standard in vitro container 1' from which the grid 6 shown in Fig. 4 and provided with its own base 11 can be removed.

[0049] List of reference symbols

Claims

Claims for a container (1) for in vitro plant cultivation, comprising a lower part (2) with a bottom (4) and a circumferential side wall (5), wherein the lower part (2) has a receiving space (8) for a nutrient medium divided into several cells (7) by a grid (6), and an upper part (3) sealing the lower part (2) in a medium-tight manner with a lid (22) and with or without a circumferential side wall (21), characterized in that the bottom (4) of the lower part (2) is divided into respective cell bottoms (18, 19) by the adjacent grid (6) and / or the grid (6) has its own bottom (11) with respective cell bottoms (18), wherein the respective cell bottoms (18, 19) are flexible and / or perforable in the direction of the cell interior and / or the bottom (4, 11) is at least partially removable from the lower part (4) or the grid (6). Container according to claim 1, characterized in that the grid (6) is integrally connected to the lower part (2).Container according to claim 1 or 2, characterized in that the base (4, 11) is substantially formed by a label (11). Container according to claim 3, characterized in that the label (11) has at least one grip tab (12) for at least partial removal from the base (2) or the grid (6). Container according to any one of claims 1 to 4, characterized in that the respective cell bottoms (18) have, in particular, (pre-)embossed thin sections (13, 14). Container according to any one of claims 1 to 5, characterized in that the cell bottoms (19) are convex and / or concave when viewed from below. Container according to any one of claims 1 to 6, characterized in that the walls of the grid (6) bounding the cells (7) have recesses (10).Container (1) for in vitro plant cultivation, comprising a lower part (2) with a base (4) and a circumferential side wall (5), wherein the lower part (2) has a receiving chamber (8) for a nutrient medium divided into several cells (7) by a grid (6), and an upper part (3) with a lid (22) and with or without a circumferential side wall (21) that seals the lower part (2) in a media-tight manner, particularly according to one of claims 1 to 7, characterized in that the side wall (5) of the lower part (2) is raised above the grid (6), wherein at least one section of the side wall (5) is reduced in height and / or reducible and / or openable. Container according to claim 8, characterized in that the at least one section of the side wall (5) is at least partially separable from the side wall (5).Container according to claim 8 or 9, characterized in that at least one section of the side wall (5) is formed by an upper, circumferential side wall part (28), which is detachably fixed to a lower side wall part (27). Container according to any one of claims 8 to 10, characterized in that a flange- and / or skirt-shaped circumferential edge (29) of the lower part (2) lies in an edge plane that runs at an acute angle to the base plane of the container (1). A method for in-vitro plant cultivation that includes the following steps: - Providing a container (1) according to any one of claims 1 to 11; - Introducing a nutrient medium into the cells (7) of the receiving chamber (8) of the lower part (2) subdivided by the grid (6); - Arranging sterile plant cell material on the nutrient medium; - Sealing the container (1) in a sterile manner; - Growing the plants (15); - Opening the container (1); - At least partial pushing of the plants (15) upwards out of the cells (7); - Grasping the pushed-out plants (15); and - Transplanting the plants (15) into a second container. Method according to claim 12, characterized in that the introduced nutrient medium solidifies into (agar) plugs (16) and / or is absorbed by porous plugs (16) inserted into the cells (7), and that the plants (15) are pushed out and grasped via the plugs (16). Method according to claim 12 or 13, characterized in that at or after opening the container (1), at least a section of the side wall (5) of the lower part (2) is reduced and / or opened. Method according to any one of claims 12 to 14, characterized in that after opening the container (1), a separate grid (6) with its own base (11) is removed from the container (1). Method according to one of claims 12 to 15, characterized in that the plants (15) or plugs (16) are pushed at least partially upwards out of the cells (7) by means of plungers (17) engaging the bottom (4, 11) of the lower part (2) or of the grid (6) removed from the container, by the plungers (17) deforming and / or piercing the respective cell bottoms (18, 19) due to their flexibility or by the bottom (4, 11) having been at least partially removed from the lower part (2) or the grid (6) beforehand.

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