Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
  • A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
  • A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
  • A01G9/029—Receptacles for seedlings
  • A01G9/0293—Seed or shoot receptacles
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
  • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
  • A01G22/60—Flowers; Ornamental plants
  • A01G22/63—Orchids
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
  • A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
  • A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
  • A01G9/029—Receptacles for seedlings
  • A01G9/0295—Units comprising two or more connected receptacles
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
  • A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
  • A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
  • A01G9/029—Receptacles for seedlings
  • A01G9/0297—Grids for supporting several receptacles
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
  • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
  • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques

Definitions

• the present invention relates to methods for cultivating epiphytes, in particular orchids and devices for use in such methods.
• Epiphytes are plants that can grow harmlessly upon other plants such as trees, and can derive moisture and nutrients from the air and rain. Therefore, many epiphytes have aerial roots. A large number of plants with aerial roots are of economic importance, mainly as ornamental plants. Orchids are of particular relevance as ornamental plants.
• the roots of plantlets obtained via in vitro cultures typically have the tendency to grow in all directions, rendering their handling difficult. Moreover, these roots are fragile and at risk of damage upon handling Adding a stress factor to the delicate transfer process by wounding of the roots is undesirable.
• Plant holders for cultivating young plants are known in the art, such as those described in NL8500603. However, these are generally used to ensure hygienic growth conditions for young plants, and do not address the above-mentioned challenges. For example, the methods described in NL8500603 are designed to further normal growth of the plant and in no way address the handling of epiphytes. Accordingly, there remains a need in the art for improved methods for cultivating epiphytes such as orchids, which mitigate at least one of the above problems.
• the present inventors have found that the cultivation of epiphytes such as orchids can be facilitated by transferring in vitro rooted plantlets from an in vitro culture to a tubular container prior to transferring the plants to a plug or loose soil for further growth.
• the tubular containers allow for pre-forming the plant roots and leaves, which may significantly facilitate subsequent transfer of the plants to a growth container. More particularly it allows handling of the plant without damaging it and makes it possible to allow transfer of the young plant to a plant pot or plug without requiring extra handling of the leaves or roots. By orienting the root tips downward, the plant will easily fit into a plug or pot. Moreover this can be achieved in a semi- sterile or sterile environment. By preforming the young plants it also becomes possible to sort plants more easily based on size. At the same time, the fact that the uniformity of the plants is increased helps to ensure that each plant is adequately positioned in the plug or pot. This allows automation of the cultivation process.
• the invention provides methods for cultivating epiphytes, comprising the step of maintaining an in vitro rooted plantlet of each epiphyte in a tubular container of a limited diameter for a time period which allows for roots and/or leaves of said plantlet to adapt to the shape of said tubular container, thereby obtaining a preformed plantlet.
• the tubular container is a container (2) with a diameter of 10-30 mm and the time period is between 3 and 21 days.
• the methods comprise the steps of transferring an epiphyte plantlet from an in vitro culture to the tubular container; maintaining the epiphyte plantlet in the tubular container until the plantlet is pre-formed; and transferring the pre-formed plantlet from the tubular container to a growth container comprising a substrate.
• the methods comprise the steps of transferring an in vitro rooted plantlet of said epiphyte to a tubular container (2) having a diameter of 10-30 mm, wherein said plantlet is positioned such that the stem and the leaf sheath is positioned within the top part of the container.
• the methods comprise the steps of transferring an in vitro rooted plantlet of said epiphyte to a tubular container (2) having a diameter of 10-30 mm and a conical shape tapering towards the bottom of said tubular container; (b) maintaining said plantlet in said tubular container for 3 to 21 days, allowing for roots and/or leaves of said plantlet to adapt to the shape of said tubular container, thereby obtaining a pre-formed plantlet; and (c) transferring said pre-formed plantlet from said tubular container to a growth container comprising a substrate.
• the transfer step is automated.
• the methods are of particular interest for plants which are a member of the family of Orchidaceae.
• the plantlet is provided with water via spraying while maintained in said tubular container.
• the plantlet comprises at least 2 roots which are between 0.5-1 cm long more particularly are at least 1 cm or at least 2 cm long upon introduction into the tubular container.
• the plantlet is introduced into the tubular container by placing the tubular container in a conductor tray (6) which comprises a duct configured such that when the tubular container is positioned the conductor tray (6), a duct of the tray is aligned with a tubular container and extends beyond the tubular container.
• the invention also provides a tubular container (2) for the propagation of epiphytes, which are characterized in that it has a diameter of 10-30 mm, a length of at least 50mm, a conical shape tapering towards the bottom of said tubular container and in that the upper rim (3) comprises a depressed portion, thereby providing the tubular container (2) with an upper lateral opening (4).
• the tubular container further comprises one or more recesses (14) in the rim of the opening (4) of the container.
• the invention further provides an apparatus for the propagation of epiphytes comprising a plurality of tubular containers (2) as described herein wherein said plurality of tubular containers (2) are configured in a linear array (1 ), laterally connected to each other.
• the plurality of tubular containers comprises at least ten tubular containers configured in a linear array.
• the invention further provides a combination of an apparatus comprising tubular containers (2) as described herein and a conductor tray (6), whereby the conductor tray comprises a plurality of parallel ducts (8) configured such that when a tubular container or the apparatus comprised of tubular containers is positioned in the conductor tray, one or more ducts of the tray are aligned with a tubular container and extend beyond a tubular container.
• the conductor tray further comprises a closing arm (13) which maintains the apparatus and/or tubular containers in position therein.
• the tray has a base for supporting the tray on a flat surface, and the plurality of parallel ducts are provided in an angle between 5° and 50° with respect to the base.
• FIG. 1 Perspective view of a particular embodiment of the apparatus comprising a linear array (1 ) of tubular conical containers (2) as described herein, and a corresponding conductor tray (6).
• FIG. 2 Perspective view of a particular embodiment of the apparatus comprising a linear array (1 ) of tubular containers (2) as described herein, positioned in a corresponding conductor tray (6).
• FIG. 3 Detailed view of a particular embodiment of the apparatus comprising a linear array (1 ) of tubular containers (2) as described herein.
• FIG. 4 Storage container (10) for storing a plurality of linear arrays (1 ) of tubular containers as described herein.
• Fig. 5 Cart (12) comprising a plurality of storage containers (10).
• FIG. 6 Perspective view of a particular embodiment of the apparatus comprising a linear array (1 ) of tubular containers (2) as described herein, positioned in a corresponding conductor tray (6) with a closing arm (13).
• FIG. 7 Front view of a young plant positioned within a tubular container; the leaves extend beyond the upper rim of the tubular container;
• FIG. 8 Front view of two examples of a plantlet before (A) and one plantlet after (B) having been pre-formed by the methods provided herein.
• heterotrophic refers to plant material that is not capable or at most weakly capable of photosynthesis. Accordingly, heterotrophic plant material requires an external carbon source such as sucrose provided in a growth medium for normal growth and development.
• autotrophic refers to plant material that is capable of photosynthesis and therefore not requiring external carbon sources for normal growth and development.
• epiphytes refers to plants which grow on other plants without taking food from the latter.
• epiphytes refers in particular, to the botanical orders of the Bromeliaceae, Araceae and the Asparagales. More particularly, it refers to members of the Orchidaceae family. Most particularly, the term “epiphytes” refers to plants belonging to the genus Cattleya, Phalaenopsis and Dendrobium.
• plantlet refers to cultured small rooted plant or seedling obtained via in vitro culturing.
• the growth medium during in vitro culture may be a liquid, solid, or semi-solid (gellike) medium.
• the plantlets used in the methods envisaged herein are obtained after having been grown on a growth medium which is a gel.
• the growth medium may comprise sucrose and agar.
• the growth medium may comprise a plant growth regulator (PGR).
• the methods envisaged herein further comprise, prior to the step of transferring the plantlet to a tubular container, the step of culturing a plantlet in vitro, i.e. typically in a culture dish in a laboratory environment.
• the plant material may go through various developmental phases before developing into a plantlet.
• the explant may form embryogenic calluses, followed by the formation of protocorm-like bodies (PLBs), which can further develop into plantlets, as is known in the art.
• PLBs protocorm-like bodies
• the methods involve the use of in vitro grown plantlets and transferring them to particular containers to direct growth of their leaves and roots.
• the methods involve using in vitro grown plantlets.
• the exact timing of the transfer from in vitro culture to the containers provided herein can differ depending on the species used, and is mainly determined by the size and number of their roots (as detailed below)
• the method described herein involves the provision of a shoot or plantlet of an epiphyte in a tubular container and allowing it to grow therein which ensures guiding and orienting of the roots and leaves of the plantlet. This allows for pre-forming the plantlet to a shape which allows for easy handling. The pre-formed plantlets are subsequently transferred to a further growth container for continued growth.
• tubular container generally refers to a tubular or cylindrical shape, i.e. a container having a diameter which is significantly smaller than the length of the container; for instance the length of the container is typically at least 2x, typically at least 3x the diameter of the container.
• tubular is not envisaged to imply any particular shape or form of the container, in that the cross-section of said container may be circular but may also be angular, such as rectangular or polygonal.
• the tubular containers as envisaged herein can be characterized by a top part (2a) and a bottom or lower part (2b). The top part is typically a section of about 2cm which may comprise a lateral opening as further described herein.
• the cross-section of the top part and the bottom part need not be the same.
• the cross-section of the bottom part may be circular, while the cross-section of the top-part is square.
• the top part and the bottom part are seamlessly integrated into one container.
• the methods provided herein may comprise a first step of transferring an epiphyte plantlet from an in vitro culture to a tubular container and a second step of maintaining the plantlet in said tubular container, thereby allowing for roots and to some extent also the leaves of the plantlet, to adapt to the shape of the tubular container thereby obtaining a pre-formed plantlet.
• the growth of the plantlet in the tubular container allows for pre-forming the roots of the plantlet vertically as the tubular container directs the roots along the longitudinal direction of the container.
• the present inventors have moreover found that this limited growth period in a tubular container can thereafter significantly facilitate the introduction of the plantlet into a growth container. More particularly it avoids the risk of damaging the roots upon introduction into the growth container.
• the increased uniformity of the methods provided herein facilitates transport and manipulation of the plantlets, thereby allowing for a higher degree of automation of the cultivation process. Indeed, the uniformity of the plants allows for automated monitoring and detection of the plantlet.
• the inventors have found that the methods allow for a more gradual transition from lab to greenhouse. Indeed, the shoots grown in vitro live on sugar but have to transition to become autotrophic when grown in a growth container. During cultivation the plantlets are grown essentially without sugars forcing the plant to become entirely autotrophic already prior to entry into the greenhouse.
• the plantlet which is introduced into the tubular container according to the methods provided herein may be obtained from a seed or explant.
• seed or explant typically, sterilized seeds or explants are used. Suitable sterilization methods are known in the art.
• explant refers to a plant tissue harvested from a donor plant, capable of vegetative reproduction in vitro on a growth medium. Such culture for vegetative reproduction is also known as “tissue culture”.
• the explant may be harvested from plant parts including but not limited to roots, root tips, rhizomes, shoot tips, stems, flower buds, and leaves.
• the plantlet will comprise one or more roots.
• the plantlet typically comprises at least 2 roots which are between 0.5-1 cm long or at least 2 cm long, preferably at least 3cm long. In these stages the roots are handled most easily.
• the plantlets comprise at least 2 roots, each of which are at least 1 cm in length. In particular embodiments, the length of the roots is less than 5.5 cm, such as less than 5cm or even less than 4.5 cm.
• the size of the plantlet upon transfer as such is not critical. Indeed, the size of the leaves in particular may vary depending on the clone. In particular embodiments, the plantlet has a size between 20mm and 150 mm (length of the longest leaf).
• the tubular container used in the present method is a hollow tube, i.e. it has a longitudinal hollow shape.
• longitudinal refers to an object having a length/width ratio of at least 2, preferably at least 3, more preferably at least 4.
• the tubular container at least in part has a conical shape, i.e. meaning that the diameter of the tubular container decreases towards one end (referred to herein as the lower end) of the container.
• a limited upper part (opposite side of the lower end) of the tubular container may be straight, while the lower part is conical.
• the entire container has a conical shape. This will prevent the plantlet from sliding down too deep into the container during transport and will moreover facilitate removal of the plantlet from the container.
• the tubular container may have an essentially conical shape, more particularly a frustroconical shape, i.e. a cone with a truncated apex.
• the tubular container may have an aperture angle, i.e. the maximal angle between two generatrix lines, between 2° and 20°, preferably between 3° and 10°.
• the tubular container is dimensioned such that it is sufficiently large such that it can accommodate the plantlet, but also sufficiently narrow (relative to the plant) such that the roots are oriented along the general direction of the longitudinal axis of the tubular container.
• the size of the tubular container can moreover to some extent be determined by the size of the growth container to which the plantlet will be transferred afterwards. Indeed, the size of the tubular container is typically at least 2 mm smaller than the smallest diameter of the growth container to which the plantlet will be transferred thereafter.
• the internal width or diameter of the tubular container in its widest measure has a diameter of at least 10 mm.
• the diameter of the tubular container at its widest point ranges between 10 mm and 50 mm, preferably between 10 mm and 30 mm, such as about 20-25 mm.
• the growth container is a conical plug with a widest diameter of about 38mm and a smallest diameter of about 28mm.
• the widest diameter of tubular container is preferably less than 28mm, more particularly 26mm or less.
• the length of the tubular container is less critical.
• the tubular container has a length between 20 mm and 300 mm, preferably between 50 mm and 200 mm, for example about 90 mm.
• the tubular container has a diameter of about 20 mm and a length of at least 50mm Such dimensions were found particularly suitable for the cultivation of orchid plantlets. Indeed these dimensions allow for the roots of the plantlets to grow vertically along the longitudinal axis of the tubular container.
• the tube has at least one open end, i.e. the upper end. This allows for introducing the plantlet into the tubular container, removal of the plantlet after pre-forming and optionally providing the plantlet with any light, water and nutrients which may be required for the plantlet's development.
• both ends (top and bottom) of the tubular container are open or provided with holes. An open bottom can allow for draining excess water, e.g. when spraying the plantlet.
• the tubular container preferably has a circular cross-section. Indeed, this optimally ensures that the growth of the leaves of the plantlets is guided without damage around a central axis which facilitates manipulation of the plantlet upon removal thereof.
• the cross-section may also have other shapes.
• the tubular container may have a cross-section which is circular, ellipsoidal, or polygonal.
• the cross-section of the tubular container may have an identical or different shape along the container's length.
• the tubular container can be divided into a top and a bottom part with a top end and a bottom end, whereby the top part is at the wider end and the bottom part is at the opposing end.
• the plantlet is typically positioned within the container such that the start of the roots is located within the bottom part, the leaves extending through the top part of the container.
• the plantlets are disposed in tubular containers such that that the plantlets' roots are positioned within the containers and the leaves of the plantlets extend above the container rim.
• Leaf sheaths are the lower parts of a leaf. More particularly, the leaf sheath is positioned below the upper rim of the container. This ensures that the leaves are guided upwards in their growth and prevents the leaves from extending horizontally from the sheath.
• the (open) top end of the tubular container comprises a rim having a depressed portion, more particularly a portion which is recessed towards the bottom of the tubular container.
• an opening can be provided in the wall of the tubular container.
• the recessed portion provides the wall of the tubular container with an upper lateral hole or opening. This can facilitate the removal of a plantlet in and from the tubular container, while still allowing for a suitable protection of the plantlet when inside the container. Accordingly, this can allow for a faster plantlet transfer, which may even be automated.
• the lateral hole or opening may further facilitate inspection and selection of the plantlets.
• the position of the opening in the tubular container is provided such that, when the plantlet is positioned therein, it corresponds to the zone of the leaves, but above the leaf sheaths.
• the sheets of several leaves often form a "stem” together, because they are wrapped around each other.
• it is of interest to ensure that the plant can be positioned in the container such that the "stem" of the leaves can be just below the opening, and the opening corresponds to a region of the leaves of the plantlet.
• the opening preferably has a height which is between 10% and 50% of the total height of the tubular container, more preferably between 10% and 30%.
• the width of the hole can vary along the height, but preferably is between 50% and 95% of the maximal width of the tubular container.
• the opening has a height between 10 mm and 10 cm, more preferably between 20 mm and 100 mm.
• the opening has an (average) width between 4 mm and 40 mm, preferably between 8 mm and 25 mm.
• the transfer of the plantlets into the tubular container and/or the removal of the pre-grown plantlet out of the tubular container is ensured either manually or in an automated way with a device comprising two arms extending through the lateral opening of the tubular container along each side of the plantlet, such as with a plier.
• a device comprising two arms extending through the lateral opening of the tubular container along each side of the plantlet, such as with a plier.
• the removal of the pre-grown plantlet is ensured with an automated device comprising two arms which are positioned to grasp the pregrown plantlet through the lateral opening of the tubular container corresponding within the region of the leaves of the plantlet.
• the rim along the top edge of the tubular container (such as but not limited to the edge of the hole or opening described above) comprises one or more recesses or other functional features which allow for manipulation of the plantlet in and out of the tubular container with a gripping device such as a plier (see below).
• tubular containers are made from materials including but not limited to plastics, glass, metal, and ceramics, but is preferably made from a plastic material.
• the tubular containers may be made from an opaque or optically transparent material.
• the plantlet is kept in the tubular container for a period sufficient to ensure that the plantlet is ready for a growth container.
• plantlet roots and leaves are preformed, i.e. for the roots adapt to the (internal) shape of the tubular container.
• the roots of the plantlets are shaped such that they follow the inner walls of the tubular container vertically downward and the leaves are shaped to grow vertically upward.
• the plantlet is kept in the tubular container for a period between 1 day and 3 weeks, more particularly between 3 and 14 days, such as between 3 and 7 days.
• a plantlet of which the roots are pre-formed using a tubular container as described herein is also referred to herein as "preformed plantlet”.
• the plantlet is preferably maintained in the tubular container under suitable climatic conditions.
• the plantlets are typically still partly autotrophic. Accordingly, while the plantlet is maintained in the tubular container, it is mainly provided with moisture.
• the plantlet may also be provided with nutrients.
• the plantlets are provided with water via spraying.
• the water which is sprayed on the plantlets comprises components such as nitrogen, potassium, calcium, iron, copper, boron, zinc.
• the water may comprise plant growth inducers or systemic pesticides.
• the tubular containers are kept at optimal climatic conditions of light (duration, quality, intensity), humidity, gas exchange, temperature (24-28 °C) to ensure further growth of the epiphyte plantlet.
• the optimal climatic conditions are known to the skilled person and may depend on the specific plant genus which is cultivated.
• the present method is described herein for the cultivation of a single plantlet, the skilled person will understand that this method is particularly suitable for mass cultivation. More particularly, the present method may involve the growth of a series of plantlets in an tubular containers as described herein. Accordingly, the methods may comprise the transfer of a plurality of plantlets obtained via in vitro culturing to a plurality of tubular containers as described herein. Typically, each tubular container is provided with a single plantlet. In particular embodiments, the plurality of tubular containers may be provided in a linear array (see further).
• the pre-formed plantlet is transferred to a container other than the tubular container described above.
• the actual size of the plantlet upon removal from the tubular container is not critical.
• the roots are less than 5.5 cm long at this stage, but this may depend on the size of the growth container to be used.
• the application thus provides plantlets, more particularly epiphyte or Phaleanopsis plantlets, which are obtainable by the methods provided herein, and are characterized in that their leaves and roots grow essentially along the same vertical axis.
• the size of the plantlets is not significantly different from that of the plantlet prior to introduction into the container.
• the preformation of the leaves and roots is significant, in that it allows easy manipulation of the plants and automated transfer into larger growth containers.
• the plantlets have roots of less than 6cm in length.
• the invention provides a set of at least 10 Phalaenopsis plantlets, characterized in that the leaves and roots grow essentially vertically along the same axis.
• the plantlets do not display roots which are positioned at an angle of more than 45° with the vertical axis extending downward from the stem.
• the plantlets have roots of less than 6 cm in length.
• the container to which the plantlet is then transferred is referred to herein as "growth container". It is characterized by the fact that it typically contains a substrate.
• the substrate can be provided as a plug (compacted substrate) or loose in a bag (see below). Growth containers and substrates suitable for cultivating epiphytes are well known in the art.
• the growth container typically is larger than the tubular container described herein above or at least has a larger diameter, thereby allowing for further growth of the pre-formed plantlet.
• the growth container may be designed for accommodating a single or multiple pre-formed plantlets.
• Typical substrates used in growth containers for epiphytes are soils which can be in powder or in fibre form.
• the loose substrate may contain a variety of materials such as tree bark (pine tree, cork), plastics (polystyrene, polyurethane foam), moss, coconut and coco products, peat, charcoal, rocks (lava rocks, pumice, perlite, rock wool), additives (sand, lime, gypsum), among others.
• the loose substrate may be highly porous or, alternatively, possess a high capillary effect. Such loose substrates allow for an optimum level of aeration and air humidity to circulate, as well as ensuring a sufficient supply of water to the plant.
• the substrate may be provided as a substrate plug.
• substrate plug refers to a composite of fibre-rich plant substrate and an adhesive, which is then shaped in a mold, thereby producing a plug.
• the fibre-rich plant substrate may contain different suitable materials, such as coconut fibres, peat, rock wool.
• Suitable adhesive may comprise a heated thermoplastic synthetic material.
• Suitable substrate plugs are known in the art. Examples include the Xtract Plug ⁇ (Quick Plug B.V), or the V-Xcel Plug ⁇ (Quick Plug B.V.), the Xcellent Plug ⁇ (Quick Plug B.V.), , Fibernet twin 3.0 or a plug such as described in EP 2327293.
• the growth containers are typically maintained in a greenhouse where the plants are further allowed to grow until marketable size.
• the apparatus comprises a plurality of tubular containers as described above, which are configured in a linear array.
• the tubular containers are provided in a rigid configuration, wherein the tubular containers are provided side by side, and parallel to each other, and preferably in a planar configuration.
• the transfer of a small plantlet to a tubular container at an early stage ensures that the roots of the plantlets are formed vertically, which facilitates manipulation of the plantlets.
• the provision of tubular containers in a linear array can facilitate manipulation and transport of the plantlets provided therein, and can therefor allow for a high degree of automation of the cultivating process.
• the linear arrays may further allow for stacking tubular containers in high densities, thereby allowing for efficiently storing and transporting plantlets.
• one or more of the tubular containers in the linear array may be removable and/or interchangeable. This allows for replacing defective containers, or moving containers from one array to another array, for example when plants within a single array.
• the apparatus may comprise a plurality of separate tubular containers, and a holder for holding the containers such that they form a linear array.
• the tubular containers may have a fixed position within the array.
• the apparatus described herein may be manufactured as a single part.
• Each of the tubular containers has an upper rim and a bottom, at opposing ends of the tubular container.
• the tubular containers are provided such that their upper rims and bottoms are aligned.
• the upper rim of the tubular containers may comprise a depressed portion, such that each of the containers is provided with an upper lateral opening or hole.
• all upper lateral opening within the linear array are provided on the same side of the tubular containers in the linear array. This facilitates providing the tubular containers with plantlets, and facilitates further manipulation of the plantlets in the containers.
• one or more of the tubular containers within the linear array may taper towards its bottom, as described above. More particularly, each of the tubular containers may have a conical shape tapering towards the bottom of the container.
• both ends of the tubular containers of the apparatus described herein may be open or provided with holes or openings, as described above.
• the rim along the top edge of the tubular container (such as but not limited to the edge of the hole or opening described above) comprises one or more recesses and/or other functional features which allow for manipulation of the plantlet upon removal of the plantlet from the tubular container and/or to handle the tubular container.
• the recesses are provided are essentially vertical (i.e. perpendicular to the longitudinal axis of the tubular container) but they may also run at a substantial angle to the longitudinal axis of the tubular container.
• the tubular container comprises at least two recesses extending from both sides of the opening, preferably in the lower half of the opening.
• a recess is between 3-7 mm wide.
• the tubular containers within the linear array may all have the same shape, or may have different shapes.
• the linear array comprises tubular containers having identical shapes.
• different linear arrays can be developed with tubular containers comprising different internal diameters for different kinds of clones (i.e. smaller and larger clones)
• the apparatus described herein allows for manipulating a plurality of containers (and plantlets contained therein) at once, thereby significantly facilitating the manipulation of plantlets according to the method described herein.
• the linear array comprises as many containers as possible. More particularly, the apparatus preferably comprises at least 5 tubular containers provided in a linear array.
• the maximal number of containers in a single linear array preferably is below 100, more preferably below 50, for example about 10 to 15.
• the apparatus described herein may be provided with one or more additional features that further facilitate automated handling of the tubular containers or plantlets contained therein.
• the apparatus described herein may further be provided with a conductor tray for facilitating the introduction of plantlets into the tubular containers of the apparatus. Accordingly, further provided herein is a combination of an apparatus as described above and a conductor tray.
• such conductor tray may comprise a plurality of parallel ducts corresponding to the shape of the tubular containers.
• the ducts of the conducting tray typically have a longitudinal shape and are configured to extend beyond the upper rim of tubular containers, when these containers are placed in the conductor tray. More particularly, the ducts are configured such that when the apparatus is positioned in the conductor tray, each of the ducts is aligned with one of the tubular containers of the apparatus.
• the ducts need not extend over the entire length of the containers but at least extend but typically extend at least along the upper part of the corresponding container. Such configuration allows for introducing plantlets into the tubular containers, via the ducts. In order to further facilitate this, the ducts are typically located above the tubular containers.
• the tray further comprises a receiving portion configured to support the apparatus within the tray.
• the receiving portion supports the apparatus by forming a ledge on which the bottom of the tubular containers of the apparatus can be positioned.
• the receiving portion comprises a clamping mechanism which fixes the apparatus at one or more edges of the apparatus, typically extending from behind the apparatus, so as to allow easy manipulation of the plantlets in and out of the apparatus.
• the conductor tray has a base for supporting said conductor tray on a flat surface, wherein said plurality of parallel ducts are provided in an angle between 5° and 50° with respect to said base. More particularly, the conductor tray may further be designed such that, when the apparatus is positioned in the receiving portion, the angle between the longitudinal axis of the tubular containers (and the ducts) with respect to the base of the conductor tray is between 5° and 50°, preferably between 5° and 20°. This can further facilitate the transfer of plantlets to the tubular containers.
• Fig. 1 shows a perspective view of a particular embodiment of an apparatus comprising a linear array (1 ) of tubular containers (2) as described herein, and a corresponding conductor tray (6).
• Fig. 3 shows a detailed view of the same linear array (1 ) as shown in Fig. 1 .
• the tubular containers In figure 3A the tubular containers have a circular cross-section, while in Figure 3B the tubular containers have a square cross-section.
• the linear array (1 ) comprises a plurality of tubular containers (2), each having an upper rim (3) and a bottom (5), at opposing ends of the containers (2).
• the tubular containers are open-ended at the top or have openings at both ends.
• the upper rim (3) of the tubular containers is provided with a recessed portion, thereby providing each container with a lateral upper opening (4). This can facilitate removal of plantlets (not shown) out of the tubular containers (2).
• the rim of the opening (4) may further comprise features such as a recess (14) for handling of the tubular container.
• each of the tubular containers (2) can be provided with a single plantlet. This may be done using a conductor tray (6) comprising a receiving portion (7) for receiving a linear array (1 ) of tubular containers (2) and a series of parallel ducts (8).
• Fig. 2 shows the same linear array (1 ) and conductor tray (6) as shown in Fig. 1 , now with the linear array (1 ) being positioned in the receiving portion (7) of the conductor tray (6).
• Each of the ducts (8) is aligned with one of the tubular containers (2), wherein the ducts (8) and tubular containers (2) have an angled orientation with respect to the base (9) of the conductor tray (6) wherein the ducts (8) are positioned higher than the tubular containers (2). More particularly, the angle between the longitudinal axis of the tubular containers (2) and the base (6) is about 10°. In this way, plantlets (not shown) can easily be introduced into the tubular containers (2), via the ducts (8).
• the conductor tray (6) may comprise a closing arm (13) which maintains the array of tubular containers (1 ) in position (Fig. 6).
• the tubular containers (2) are rigidly fixed to each other, thereby allowing for manipulating and transporting all tubular containers (2) within the linear array (1 ) at once.
• a storage container (10) for storage and transport, several linear arrays (1 ) may be placed in a storage container (10) as shown in Fig. 4, wherein the linear arrays form different rows within the storage container.
• the container (10) may further be provided with a lid (1 1 ) for protecting the plantlets and/or allowing for a controlled atmosphere within the storage container (10).
• the lid (1 1 ) may be provided with holes or transparent regions such that the plantlets in the tubular containers (2) can be provided with light.
• Multiple storage containers (10) comprising the tubular containers (2) may be stacked onto a cart (12) as shown in Fig. 5, thereby allowing for an efficient transport of plantlets.
• Fig. 7 shows a linear array (1 ) comprising a plurality of tubular containers (2).
• Each of the tubular containers comprises an upper part and a lower part,
• the tubular containers are open-ended and comprise an upper rim (3).
• the upper part further comprises, a lateral opening (4).
• Plantlets (15) are disposed in the tubular containers (2) such that the roots are placed within the bottom part of the tubular container and the leaves of the plantlets extend through the upper part above the rim of the tubular container. In other words, the sheath of the leaves or stem of the plantlet (15) is located below the upper rim (3) of the tubular container (2) in which the plantlet (15) is positioned.
• Fig. 8 shows examples of plantlets (15) having leaves (16), roots (17), and a stem (18).
• the plantlets in panel A are obtained by microculture and have roots and leaves which grow in different directions, more particularly the leaves and roots are not positioned vertically along the same axis; in particular, some roots of the plantlets in panel A grow at an angle of more than 45°, or even more than 90° from the vertical axis extending downward from the stem of the plantlet.
• the plantlet in panel B (15) is an example of a plantlet which has been pre-formed by means of a tubular container according to particular embodiments of the methods provided herein.
• the plantlet's (15) roots (17) were positioned in the tubular container, the plantlet's (15) stem (18) was positioned below the upper rim of the tubular container, and the plantlet's leaves (16) were allowed to extend beyond the upper rim (3).
• the plantlet was maintained in the container for a period of 3-7 days. As a result, the leaves and roots of the plantlet grow along the same vertical axis. In this way the plantlet was formed in a shape which allows easy handling and ready automation.

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• Life Sciences & Earth Sciences (AREA)
• Environmental Sciences (AREA)
• Botany (AREA)
• Engineering & Computer Science (AREA)
• Biotechnology (AREA)
• Cell Biology (AREA)
• Developmental Biology & Embryology (AREA)
• Cultivation Receptacles Or Flower-Pots, Or Pots For Seedlings (AREA)
• Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

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Citations (5)

• 2015
  • 2015-06-30 NL NL2015052A patent/NL2015052B1/en not_active IP Right Cessation
  • 2015-07-30 BE BE2015/5486A patent/BE1023173B1/nl not_active IP Right Cessation
• 2016
  • 2016-06-30 TW TW105120826A patent/TW201707558A/zh unknown
  • 2016-06-30 WO PCT/EP2016/065325 patent/WO2017001571A1/en active Application Filing

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