Method and device for propagating living organisms

Abstract

The invention relates to a method for propagating living organisms, in particular plants, fungi, insects and / or aquacultures having the following steps: A Providing an explant (120) of a living organism; C In-vitro incorporation of the explant (120) into a medium to form an in-vitro cell network; D1 Cloning the in-vitro cell network with formation of a cloning starting material; and / or, E Rooting and / or incubating the cloning starting material, wherein in the course of the method a determination of a living organism species and / or a living organism part from which the explant was provided is carried out as a dataset, and a control of at least one process parameter for setting the growth conditions in the preceding and / or subsequent steps is carried out on the basis of the determined dataset; and an apparatus (100) for carrying out the method.

Description

[0001] METHOD AND DEVICE FOR REPROPAGATION OF LIVING BEINGS

[0002] The present invention relates to a method and a device for propagating living beings.

[0003] Living organisms such as insects, aquacultures, fungi, or plants are often produced on a large scale today to meet global demand. However, such production is often characterized by manual steps, leading to production bottlenecks or massive labor requirements. Some sub-steps are completely automated, as disclosed in DE 10201601068A1. This describes an automated cloning step for ornamental plants, but without taking the origin of the explant into account. DE 202022105678 U1 discloses the detection of diseases in leaves. WO 2022253523A1 discloses the semi-automatic propagation of plants. DE 10055907 discloses a container for cultivating zooplankton and phytoplankton, and DE 202021100695 discloses an aquaculture system.

[0004] All of these systems only allow the automation of partial steps, and none of them covers the entire propagation cycle of a living organism, which in plants, for example, includes the extraction of explants, the in vitro cultivation of a new plant, the cloning of said plant, and the rooting of the respective plant. In particular, the currently known methods lack a statistical and / or combinatorial approach to determine or adjust optimal propagation and / or growth conditions. Furthermore, no method is known for testing successfully cloned organisms for mutations or other stress factors.

[0005] Based on the aforementioned prior art, the object of the present invention is to provide a method and a device for creating automatic and optimized reproduction and / or growth conditions for living beings.

[0006] The present invention solves this problem by providing a method having the features of claim 1 and an apparatus having the features of claim 10.

[0007] A method according to the invention for the propagation of non-human living beings comprises at least the following steps:

[0008] A Provision of an explant of a non-human organism; C In vitro uptake of the explant into a medium to form an in vitro cell cluster, preferably an embryo, seedling, larva, egg or spore.

[0009] D1 Cloning of the in vitro cell network to form a cloning starting material and / or

[0010] E Rooting or incubating the cloning starting material.

[0011] Optionally, in a step F, one or more phenotypic parameters and / or growth parameters can also be determined.

[0012] According to the invention, in the course of the method, the type variety is determined, which can be, for example, the food variety, e.g. the plant variety and / or the food part, e.g. the plant part, from which the explant was provided, which is carried out as a data set and a control of at least one process parameter for setting the growth conditions in the preceding steps is carried out on the basis of the determined data set.

[0013] The control and thus the promotion of growth can thus be individually adapted to the type of organism, for example, the plant or the type of plant part. This enables particularly effective, automated and resource-conserving reproduction of organisms.

[0014] In the context of the present invention, the term “living beings” preferably includes the classes of insects, aquacultures, plants and / or fungi.

[0015] Further advantageous embodiments of the method according to the invention are the subject of the subclaims.

[0016] It is advantageous for the creation of controlled and uniform initial conditions if the process includes a step B between steps A and C as cleaning and / or sterilization of the explant.

[0017] The method may furthermore advantageously comprise a preservation of the cell composite after step D1 or simultaneously with step D1.

[0018] The phenotypic parameter according to step F advantageously comprises or corresponds to one or more of the following variables: morphology, dimension, color, electromagnetic absorption, growth rate, transpiration molecules, respiratory molecules, excretion molecules, and / or mobility. Mobility is particularly interesting in the case of insects and aquaculture. The determined phenotypic parameter, in addition to the data set regarding the organism species and / or organism part, is considered as a decision parameter when adjusting, in particular controlling or regulating, the process parameter.

[0019] The respective process parameter to be set includes one or more of the following variables:

[0020] • Light intensity and / or light spectrum

[0021] • Humidity and / or air temperature

[0022] • Composition of a cleaning agent and / or type of sterilizing agent

[0023] • Concentration of one or more nutrients in a nutrient medium, in particular hormone concentration in a nutrient medium and / or

[0024] • Temperature, viscosity and / or ion concentration of a nutrient medium.

[0025] • Additionally for fungi, plants and insects the humidity and composition of the atmosphere

[0026] • Additional for plants the droplet size of the particles of the air humidity

[0027] • Additionally for aquaculture the nutrient medium and / or water temperature

[0028] The automated process may comprise, in a step G, placing the embryos, seedlings, larvae, eggs or spores into suitable transport containers.

[0029] The process is carried out in an aseptic atmosphere, at least from step B to step E, and preferably until step G. This allows plants or other living organisms used in the pharmaceutical industry to be cultivated.

[0030] In particular, the steps AG are carried out without manual intervention by a person. In particular, physical contact with people is completely avoided in the aforementioned procedures.

[0031] The method can comprise, as a sub-step of step G, for example, a selection of a packaging material and / or a packaging process depending on the data set relating to the plant variety and / or plant part. Individual plants, e.g., nightshade plants, can be sensitive to direct sunlight. These should be packaged, for example, in a tinted film. The packaging material can further protect against atmospheric influences, such as foreign gases or toxic gases, e.g., O2 for plants or CO2 for insects, or simulate a natural environment, such as nutrients in the water for aquaculture.

[0032] The invention further relates to an apparatus for carrying out the method according to the invention. The apparatus can have a plurality of successively arranged compartments, wherein at least one or more of the compartments are designed as a transport device or wherein the compartments are connected to one another by transport devices.

[0033] Furthermore, according to the invention, the apparatus has at least one compartment that is maintained under aseptic conditions. This enables automated and sterile processing of the organisms.

[0034] It is advantageous if at least one compartment has a sensor arrangement, in particular an image capture system, for determining the species and / or part of the organism. This can, for example, be an image capture system with subsequent evaluation regarding dimensions, color, or mobility, or a combination of these for the recognition, also with the aid of algorithms, of parts of the organism, such as a stem, a flower, a fruit, a head, legs, antennae, or a tail.

[0035] Furthermore, it is advantageous if at least one or more compartments are designed as a segmented chamber wheel with a plurality of cell chambers arranged radially around a rotation axis. The arrangement in cell chambers allows each chamber in a compartment to have its own microclimate or nutrient supply protocol. In this way, several different living organism species or the same species with different microclimates and / or nutrients can be bred in one and the same compartment. Taking into account the time required to carry out each process step, the time required to carry out each process step is used optimally. For example, a plant genus can be placed in a compartment with different or identical microclimates and different or identical nutrients in order to statistically and / or combinatorially develop the optimal growth conditions.

[0036] Further advantages, features, and details of the invention will become apparent from the following description, in which several embodiments of the invention are explained in more detail with reference to the accompanying figures. Those skilled in the art will conveniently consider the features disclosed in the figures, the description, and the claims individually and combine them into useful further combinations. They show:

[0037] Fig. 1 schematic representation of an embodiment variant of a device according to the invention for carrying out a method according to the invention.

[0038] Fig. 1 shows an apparatus 100 for developing and covering the entire industrial propagation of living organisms. The apparatus and the underlying method preferably cover the aseptic, industrial propagation of living organisms with the steps

[0039] (A) Illustration of explants;

[0040] (B) cleaning and / or sterilization of explants,

[0041] (C) in vitro uptake of explants into a medium,

[0042] (D1 ) Cloning of in vitro cell clusters and / or

[0043] (D2) Preservation of cloning starting material,

[0044] (E) Rooting and / or incubation of the starting material.

[0045] (F) Determination of phenotypic parameters and / or

[0046] (G) Placing the embryos, seedlings, larvae, eggs or spores in suitable transport containers.

[0047] In particular, the apparatus and the method are agnostic with regard to the type of living organisms, e.g. it is irrelevant whether pharmaceutically interesting plants, other crops, food, fungi, algae, insects, larvae, eggs, small crustaceans and / or small fish are processed.

[0048] The process is preferably carried out aseptically, at least from step (B), with a sterility of SAL 10-6 CFU according to ISO 11137. For example, sterility can be achieved by means of suitable sterilization processes and agents, such as hydrogen peroxide, ozone, ethylene oxide, perchloroacetic acid, UV-C, eBeam, gamma radiation, ethanol, or isopropanol.

[0049] The sub-steps (A) to (F) can be carried out sequentially or in parallel with at least one station of the apparatus 100 per sub-step.

[0050] The process is supported by software, e.g., algorithms, self-learning AI, and / or supervised AI, to learn from literature, interpret, and independently test probabilities of promising process sequences, and / or through specific and / or planned statistical and / or combinatorial experiments. In particular, the aspect of literature interpretation as an input factor allows the software to continually acquire more process knowledge about different explants, protocols, nutrients, microclimates, and procedures. This can lead to specific experiments and / or processes not being performed from the outset, or to experiments being deliberately performed due to a lack of data.For example, it may be that a nutrient or toxin that is not yet sufficiently researched is always tested in certain vulnerable organisms, whereas another nutrient, for which there is sufficient knowledge about its biological inertness, is omitted in specific organisms.

[0051] The apparatus according to the invention only allows a high-throughput screening of possible process factors in steps A - G in combination with suitable control and interpretation software.

[0052] The procedural steps in detail:

[0053] Step A

[0054] From now on, plants will be used as an example in the descriptions to represent the living beings listed.

[0055] The suitable apparatus 100 for step A has a first compartment 20 into which the plants can be introduced via a suitable lock 7 or opening. Such a compartment can be designed, for example, as a safety workbench for biological work or as an isolator workbench.

[0056] The plants can be introduced into a chamber 23, for example, in a pot or similar containers. These plants, also called mother plants 110, can be picked up from these containers by a suitable conveyor mechanism 22. The mother plant is first positioned by the transport device 4 in an inspection station 23, where it is visually measured using suitable measures to determine the type of plant or the type of specific plant parts.

[0057] The plant parts to be identified are preferably leaves, stem, buds, flowers, fruits, nodes, rhizomes, roots, and / or root tips. In the case of mushrooms, these are the cap, gills, collar, stalk, sheath, or tuber. In the case of fish and fish larvae, these are the head, dorsal fin, caudal fin, gills, eyes, or, in the case of translucent fish and larvae, individual organs.

[0058] To determine the type of plant and / or plant parts, the apparatus

[0059] 100 has a camera system 1. Preferably, the camera system 1 is designed for 3D image capture to determine the type of plant and / or plant parts. For this purpose, the camera system 1 can also have two or more cameras, which allow capture from different angles.

[0060] Particularly preferably, a corresponding camera system 1 is confocal and / or equipped with a sensor and / or evaluation unit for transmitting and / or receiving measured values ​​in the wavelength ranges of UV, VIS, NIR, and / or IR radiation.

[0061] Alternatively or additionally, the camera system may comprise a sensor and / or evaluation unit 2 for carrying out an imaging method in the form of stereoscopy, LiDAR, ultrasound, time-of-flight, laser and / or one or more other imaging methods.

[0062] Using suitable portioning means, e.g., grippers, scalpels, punches, and / or a combination thereof, the desired plant parts with a specific size, weight, and / or shape are explanted from a mother plant 110 as an explant 120. This is done by a removal device 3. The inspection station 23 and the removal station intended for removal can be one and the same station. Thus, after inspection, portioning can take place without moving the mother plant. This is particularly advantageous for fragile organisms, such as larvae or eggs, in order to avoid further mechanical stress on the organism.

[0063] However, in order to accurately determine plant data, it may be advantageous to provide a spatial separation between the two stations, i.e. the sampling station and the inspection station, in order to avoid contamination of the inspection station.

[0064] The advantage of sampling or explantation after inspection is that the results of successful explantation and subsequent steps are comparable in terms of in vitro uptake in relation to quantity, shape, and / or mass. This allows conclusions to be drawn as to which part of the plant is most successful in terms of explantation. For example, it can be determined whether increasing the explant mass has a positive effect on further processing into an in vitro culture.

[0065] In particular, this standardized explantation step allows for multiple explantations from the same plant in the compartment at the same or different times, thus allowing for the determination of a mutation rate in subsequent steps using suitable analytical methods. Suitable methods here primarily refer to gene sequencing, such as PCR, NGS, or similar methods, and are possible in all subsequent steps. The explants 120 can then be arranged in an array 6 comprising multiple storage locations.

[0066] Step B

[0067] An explant 120 after step A is then washed and / or sterilized in step B using cleaning media 25 and / or other agents known per se. For this purpose, a cleaning chamber 24 is provided into which the explant(s) are transported further after their removal by a transport device 4. The type and / or dosage of the cleaning media and / or the duration of the cleaning can be tailored to the type of plant or the type of plant parts based on the determination in step A. For example, roots often contain inorganic contaminants such as sand or the like, while leaves may contain aphids, for example. Accordingly, the selection of the cleaning agent and the duration of the cleaning can be adjusted based on the measurement data from step A.

[0068] This cleaning and / or sterilization step preferably achieves external sterility of the explant, but existing biological contaminants, such as viruses, fungi, bacteria, spores, and prions, are still present in the explant. Therefore, cleaning has the special function of allowing, for example, in vitro uptake of the plant material into the subsequent compartment 30 under external and internal sterile conditions.

[0069] Sterilization can be performed either wet-chemically or alternatively or additionally by irradiation. For example, sterilization can be performed by gamma irradiation. Further transport to the subsequent compartment 30 is carried out via transport device 4.

[0070] Step C

[0071] The downstream compartment 30 is designed in Fig. 1 as a segmented chamber wheel with a plurality of cell chambers 31. The cell chambers 31 can each be individually sterilized. The explant 120 arranged therein or the arrangement of a plurality of explants 120 arranged therein are accessible via a robot arm 32 and can thus be moved. The compartment 30, as well as all downstream compartments 40-80, can analogously have a corresponding robot arm 32. Furthermore, the compartments can be placed individually or as a whole in an atmosphere, e.g., an aseptic atmosphere. For optimal accessibility and transfer of the explants 120 in the cell chambers 31, the segmented chamber wheel can be designed to rotate around a central axis. This can also apply analogously to all downstream compartments.

[0072] Finally, one or preferably each compartment 30-80 has a sensor arrangement 9 for determining plant data. This can preferably be an optical sensor, such as a camera system comprising one or more cameras, which, for example, detect the shape and color of the explants and control a nutrient dosing system 10 likewise arranged in the compartment. The nutrient dosing system can be adjusted based on the sensor data from the sensor arrangement 9. This particularly concerns the adjustment of the concentration of one or more nutrients and / or hormones in a nutrient medium added by means of the nutrient dosing system 10. The residence time of the explant(s) 120 in the respective compartment 30-80, light intensity and / or the atmosphere, in particular the air humidity, oxygen content, CO2 content and / or droplet size of the moisture in the air, can also be adjusted according to the sensor data.

[0073] In particular, a comparison is made between the target and actual values, for example of the plant colour, the anticipated size of the plant or individual plant parts after a certain growth period and / or the concentration of plant emissions. If the actual value lies outside a limit value or a target value range, a first variable, e.g. the light intensity, can be adjusted. If this change does not result in a significant change in the actual value within a certain time interval, another variable, e.g. the hormone concentration in the nutrient medium, can be adjusted. If this also does not result in a significant change in the actual value within a certain time interval, a further variable, e.g. the air humidity or the phosphate content of the nutrient medium, can be adjusted.A statistical and / or combinatorial approach can also be used to adjust the variables in order to execute time-critical processes and / or the processes in general simultaneously and thus in a time-saving manner.

[0074] The extent of the time interval during the measurement and the decision of prioritization as to which variable is changed first, second, third, etc., can be made by an artificial intelligence, which, for example, uses empirical values ​​from previous growth behavior of other plants in the apparatus according to the invention to determine the aforementioned data.

[0075] Preferably, a measurement of the size of the explants 120 within a cell chamber 31 of a compartment 30-80 is carried out after a predetermined time interval. This eliminates the need to monitor each cell chamber 31 separately; instead, monitoring or determining the sensor data can take place in a specific rotational position of the cell chamber 31 of a compartment 30-80.

[0076] The suitable apparatus 100 comprises supply devices 8 as part of the second compartment 30, through which the explants 120 from the first compartment 20 are supplied with biological material. In this step, the externally sterile explants are also generated internally sterile, e.g., free from biological contamination, through suitable in vitro uptake and propagation via the route of direct or indirect embryogenesis. For this purpose, the explants 120 introduced into compartment 30 must preferably have the same mass, and both routes of embryogenesis are initiated in parallel by the apparatus 100 with all explants from the various plant parts.

[0077] Cell clusters, egg cells, microspores, somatic cells, or callus are particularly suitable as explants. In particular, the specific cell cluster called callus is characterized by direct embryogenesis through the formation of the globular follicle and the subsequent steps known in the literature to the gymosperm, monocot, and / or dicot.

[0078] For example, and / or in addition, identifying parts as root meristem, hypocotyl, shoot meristem, and / or cotyledon can be advantageous. These parts, or rather their development, can be used to calculate viability and proliferation rate, which in turn can be calculated as a key figure to make an (automatic) decision regarding the explant and protocol that will lead to the fastest and most reliable development of a viable and reproducible embryo.

[0079] In indirect embryogenesis, the explant undergoes dedifferentiation, forming the cotyledon, as described in the literature. Both embryogenesis pathways require testing of various hormones on different culture media, sometimes with multiple changes of hormones and / or culture media. This particular task is preferred for two reasons. a) By testing various culture media and hormones (also called hormone squares), the optimal mixtures, concentrations, processes, and / or media can be found quickly and automatically through combinatorics. This aspect is conveniently and quickly fulfilled with the present invention of the apparatus (100). b) By repeatedly transferring callus cell clusters or other developmental and maturation stages of living organisms resulting from the biological process to sterile media, the outgrowth of biological contamination can be achieved.

[0080] Thanks to the automation of the process using the apparatus 100, transfer to sterile media can be carried out multiple times and with high reproducibility during development and / or within a development stage.

[0081] An exception to this are pure root cultures, which are grown in a bioreactor and use a nutrient liquid as the medium. Although the principle remains the same regarding nutrient exchange, for root cultures and / or aquaculture, it is not the medium exchange that is relevant, but rather the added nutrients and other environmental parameters, such as the temperature of the medium, dissolved gases, or movement of the medium.

[0082] In particular, compartmentalization of the nutrient media in the dosing device 10 is recommended for this step in order to prevent cross-contamination.

[0083] Since embryogenesis potential is genotype-dependent, a route for preparation via explant or undifferentiated cells must be pursued. Compartment 30 of Apparatus 100 is helpful in this regard, as multiple genotypes can be processed simultaneously.

[0084] A rapid selection of embryogenesis cultures and non-embryogenesis cultures is easily carried out by means of the present invention, since the explants, undifferentiated cells and / or cells can be tracked and documented quickly, in a standardized manner and / or after short time intervals of a few minutes to a few hours using photographic and / or spectroscopic methods.

[0085] A further advantage of compartment 30 of the apparatus 100 according to the invention is that chemicals, e.g., hormones, cytokines, and vitamins, can be tested quickly, precisely, and statistically and / or combinatorially to test and determine the embryogenetic competence of a sample. These can be coordinated with the inspection performed in step A. For example, by preparing, verifying, and monitoring a concentration series of one or more vitamins using spectroscopic methods, embryogenetic competence can be determined as a factor of time and concentration.

[0086] This also allows chemical, biochemical, and molecular events to be detected, thus influencing parameters of a sample's embryogenetic competence. For example, hormone screening and / or hormone squaring can be used to determine the optimal hormone to maximize embryogenesis in terms of quantity and speed.

[0087] In particular, the zygotic and somatic embryogenesis pathways can be simultaneously determined, tested, documented, and / or influenced using Apparatus 100. For example, by simultaneously tracking both embryogenesis pathways, not only can the time required for embryogenesis be reduced, but the better pathway, in terms of successful embryogenesis, can also be simultaneously tested in a shorter time.

[0088] Embryo propagation and / or cloning can also be performed in this step, which has the advantage that cell clusters are formed rather than sprouts or seedlings, which might react to the stress of cutting, cloning, and / or propagation through unwanted mutation. Thus, this step C of embryogenesis and / or in vitro cloning is the step in which it is determined which factors need to be controlled for successful embryogenesis and / or in vitro cloning. With successful embryogenesis and / or in vitro cloning, the embryo can be allowed to develop in the next step.

[0089] Step D

[0090] Step D can be divided into two substeps, D1 and D2, which can be performed sequentially or in parallel, with D1 preferably always being performed. A third compartment 40 is provided for this purpose.

[0091] In step D1, direct cloning from an embryo and / or other meristem tissue is undertaken by either the zygotic or somatic route.

[0092] Since the embryos from step C can already be available individually, there is no need for propagation or cloning from a later offspring, as is done in other procedures. Cloning from an embryo is therefore merely the outgrowth of the embryo and not, in the truest sense, cloning the embryo.

[0093] By developing the embryo into a seedling under optimal conditions, mutations are also reduced. In this compartment 40, the optimal protocol for raising a seedling from an embryo can be developed quickly and precisely using combinatorics, DoE, neural networks, artificial intelligence, and / or statistics and analytics. This substep D1 is integrally included in the method according to the invention and allows for individual measurements of an embryo to produce a seedling. Again, using suitable mathematical methods, e.g., combinatorics, time series analysis, DoE, multiple identical and / or different embryos can be measured and analyzed simultaneously. This particularly reduces the time required to develop a process protocol and also generates in-depth knowledge of derived processes and / or molecular and / or biochemical events.For example, by adding nutrients during seedling development, one or more nutrients can be varied to determine whether there is a maximum or minimum in one or more phenotypic parameters, such as the growth rate of a seedling. Furthermore, by varying the embryos but keeping the concentration and / or timing of addition of one or more nutrients constant, similarities in processes can be determined, which serves as a basis for decisions regarding the application and / or omission of nutrients in the algorithm for future embryos.

[0094] In the optional step D2, an embryo and / or other meristem tissue is encapsulated into a synthetic seed. This occurs in a fourth compartment 50. Encapsulation has the advantage that preservation can be achieved easily and without the need for deep freezing or maintenance cultivation of a mother plant. Step D2 also allows for such encapsulation for spores, whereas deep freezing is advantageous for insects, larvae, and eggs.

[0095] Using suitable mathematical methods, e.g., combinatorial, DoE, or time series analysis, the optimal encapsulation process with different substances, e.g., agar agar, gelatin, can be achieved simultaneously, in a standardized, and timely manner. For example, in well plates, the combinatorial addition of various encapsulation substances, such as agar agar, gelatin, and / or xanthan, can be used to determine the encapsulation achieved using visual detection methods as a function of time and morphology and compare it with target values. The combinatorial mixture can be adapted, controlled, and / or calculated based on the previously obtained results until a target criterion, such as less than 5 minutes and a round to oval shape, is achieved during encapsulation.

[0096] Compartment 50 also serves as a storage facility for these synthetic seeds and is accordingly controlled with regard to atmospheric conditions.

[0097] Each of the compartments of the device 100 according to the invention is individually controlled with respect to atmospheric or medium conditions, nutrient supply, and / or light irradiation. Control is carried out, in particular, based on the identification in step A or further measurements in subsequent steps. Step E

[0098] Step E requires a separate compartment 60 for rooting and / or incubating the embryos and / or meristem tissue preserved in step D2 and / or produced in step D1. This compartment 60 can perform the rooting of individual starting materials spatially separated but simultaneously.

[0099] In this case, suitable mathematical methods, such as combinatorics, DoE, or time series analysis, can be used to test different chemicals and / or solvents on different culture media in order to quickly develop an optimal rooting protocol. For example, increased rooting through hairy roots can be considered optimal, which can be detected qualitatively, semi-quantitatively, or quantitatively using a camera system with image analysis.

[0100] Using suitable analytical methods, e.g., measuring the concentration of nitrates, phosphates, and the like, preferably using ion-selective electrodes or light intensity and flow rate, measurement points can be established that allow conclusions to be drawn about possible molecular and / or biochemical events using logic, statistics, or combinatorics. Imaging techniques, constituent and / or gas-phase analysis are particularly suitable for this purpose. This compartment 60 is directly connected to two other compartments 70 and 80.

[0101] Step F

[0102] Step F is preferably carried out in a separate compartment 70 and serves to measure individual seedlings for phenotypic growth parameters. This compartment can also be designed as a single station. This compartment 70 can, for example, take the form of one or more cabinets mounted on a rotating plate.

[0103] By measuring individual or sum parameters of seedlings from step E as individual seedlings and / or in groups, an optimized growth protocol can be developed using suitable sensors and control of the atmospheres around the seedlings.

[0104] In particular, sensors are designed to measure air pressure, atmospheric gases, light intensity and brilliance, temperature and humidity, volatile organic components, the optical and dimensional surface area of ​​the plant, and / or the flow of nutrients to and from the plant. An aeroponic system is used for this purpose, with nutrients being delivered to the plant as a mist.

[0105] In particular, optical measurements of the plant in the electromagnetic spectrum at wavelengths from 100 nm to 1 mm are of interest for phenotypic parameters. These wavelengths provide different information about the plants and are therefore important for monitoring the plant's growth process.

[0106] UV radiation (100 nm - 400 nm) is interesting for detecting various surface effects, such as the formation of secondary metabolites and toxins, such as furocoumarin, on the plant, which can indicate nutrient excess or deficiency.

[0107] The wavelengths of 400 nm - 800 nm in the visual range indicate color changes on the leaf, which, for example, indicate a calcium deficiency.

[0108] Wavelengths from 700 nm - 2500 nm allow easy penetration into the surface of the plant and can thus measure various aspects, such as water content in the leaf or at the stem in the plant.

[0109] In addition, this wavelength range, e.g. near infrared (NIR) and infrared, can be used to detect molecules in the gas phase that are secreted by the plant.

[0110] In particular, NIR is interesting for measuring the phenotypic parameter of respiration and transpiration of the plant, since NIR can also be used to penetrate into matter, usually requires no sample preparation and / or to measure molecules with very low dipole moment, such as oxygen, carbon dioxide, nitrogen and similar molecules.

[0111] Infrared (IR) is used to detect mostly organic molecules and water and can also be used for the gas-phase detection of organic volatile molecules (VOCs). These VOCs occur as parameters in the plant during various phenotypic processes and are secreted by different organs of the plant to signal a specific status or stage of plant growth. For example, during the ripening process of fruits, specific VOCs are secreted, which can be detected in this way. This allows the precise determination of the harvest time. Using suitable mathematical models, all sensor data can not only be evaluated and / or monitored, but also information can be derived from it that allows a prediction of phenotypic results.In particular, time series analyses, neural networks, and / or DoE and derived prognostic methods and algorithms offer advantages when it comes to understanding biological processes in plants through phenotypic characterization.

[0112] This step F can also be carried out in isolation and does not necessarily have to be used for industrial propagation.

[0113] Step G

[0114] In step G, the industrially produced and / or propagated seedling is placed in sterile packaging in compartment 80, again under sterile conditions. After the seedling is placed in each packaging unit, at least two packaging units are palletized together and prepared for shipping.

[0115] Steps A through G, with the exception of step F, must be performed under sterile or aseptic conditions for successful, automated, industrial propagation of living organisms, especially plants. The optional step D2 can be used to maintain cultures without relying on a mature plant, a so-called mother plant.

[0116] The compartment 80 may comprise a packaging unit 13 for a plurality of packaging variants depending on step A. This includes, for example, packaging in an aseptic atmosphere with a metal-coated film material or a metal foil to prevent gas exchange between the interior atmosphere of the package and the environment.

[0117] If an aseptic atmosphere is not required for the processed plant, the packaging unit can be made of transparent plastic for easier plant identification. This material at least prevents microorganisms from coming into contact with the plant.

[0118] Alternatively, a simple cardboard material can be selected for packaging. This can be used for particularly resistant plants or where requirements do not require hermetic protection. Accordingly, the packaging unit has a magazine made of different packaging materials, with the selection of the packaging material depending on the identification or other sensor data collected during the process.

[0119] Alternatively or additionally, the packaging conditions can also be adjusted based on the various sensor data. The compartments are advantageously connected to each other by transport devices, e.g., conveyor belts, robot arms, or the like, enabling an automated process. Manual interventions in the manufacturing process can be performed in the apparatus 100, preferably via service robots or via application in a glove box.

[0120] In an optional final station 14, the produced and packaged living beings can be palletized.

[0121] Reference symbol

[0122] 1 camera system

[0123] 2 Evaluation unit

[0124] 3 Removal device

[0125] 4 Transport device

[0126] 6 Arrangement of storage spaces

[0127] 7 Lock

[0128] 8 Nutrient dosing system

[0129] 9 Sensor arrangement

[0130] 13 packaging units

[0131] 14 palletizing unit

[0132] 20 compartments

[0133] 22 Funding mechanism

[0134] 23 Chamber / Inspection Station

[0135] 24 cleaning chamber

[0136] 25 cleaning media

[0137] 30 compartments

[0138] 31 cell chamber

[0139] 32 robot arm

[0140] 40 compartments

[0141] 50 compartments

[0142] 60 compartments

[0143] 70 compartments

[0144] 80 compartment

[0145] 100 equipment

[0146] 110 mother plants

[0147] 120 explants

Claims

Patent claims 1. A process for the propagation of living organisms, in particular plants, fungi, insects, and / or aquaculture, comprising the following steps: A providing an explant (120) of a non-human living being; C In-vitro uptake of the explant (120) into a medium with formation of an in-vitro cell network, D1 Cloning of the in vitro cell network to form a cloning starting material and / or E Rooting and / or incubating the cloning starting material, characterized in that in the course of the method a determination of a living being location and / or a living being part from which the explant was provided is carried out as a data set and that a control of at least one process parameter for setting the growth conditions in the preceding and / or subsequent steps is carried out on the basis of the determined data set.

2. Method according to claim 1, characterized in that the method comprises a step B between steps A and C as cleaning and / or sterilization of the explant (120).

3. Method according to claim 1 or 2, characterized in that the method comprises preserving the cell network after step D1 or at the same time as step D1 4. Method according to one of the preceding claims, characterized in that after step E, in a step F, a determination of one or more phenotypic parameters takes place, wherein the phenotypic parameter comprises one or more of the following variables: morphology, dimension, color, electromagnetic absorption, growth rate, transpiration molecules, respiratory molecules, excretion molecules and / or mobility.

5. Method according to one of the preceding claims, characterized in that the determined phenotypic parameter is taken into account when setting the process parameter.

6. Method according to one of the preceding claims, characterized in that the process parameter comprises one or more of the following variables: • Light intensity and / or light spectrum • Humidity and / or air temperature • Droplet size of the particles of the air humidity • Composition of the atmosphere Composition of a cleaning agent and / or type of sterilizing agent • Concentration of one or more nutrients in a nutrient medium, in particular hormone concentration in a nutrient medium and / or • Temperature, viscosity and / or ion concentration of a nutrient medium.

7. Method according to one of the preceding claims, characterized in that the method comprises, in a step G, placing the cell assembly into a suitable transport container.

8. Process according to one of the preceding claims, characterized in that the process from step B to step E, preferably to step G, is carried out in an aseptic atmosphere.

9. Method according to one of the preceding claims, characterized in that the method comprises selecting a packaging material and / or a packaging process depending on the data set.

10. Apparatus (100) for carrying out a method according to one of the preceding claims, characterized in that the apparatus has a plurality of successively arranged compartments (20, 30, 40, 50, 60, 70, 80), wherein at least one or more of the compartments are designed as a transport device or wherein the compartments (20, 30, 40, 50, 60, 70, 80) are connected to one another by one or more transport devices (4).

11. Apparatus according to claim 10, characterized in that at least one compartment has a sensor arrangement (9), in particular an image capture, for determining the type of living being and / or the part of the living being.

12. Apparatus according to one of the preceding claims 10 or 11, characterized in that at least one or more compartments (30, 40, 50, 60, 70, 80) as a segment chamber wheel, with a plurality of cell chambers (31) which are arranged radially around an axis of rotation.

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