AUTOMATED SEED SAMPLE AND METHOD REMOVAL SYSTEMS TO REMOVE SEED FABRIC SAMPLES

专利摘要:
automated systems to remove seed tissue samples and related methods. It is a seed sampling system that has an automated seed loading assembly that includes a seed container and is operable to select seeds from a plurality of seeds in the seed container. The system also includes an operable automated seed sampling assembly to remove tissue samples from selected seeds and an operable automated seed transport assembly to transfer selected seeds from the seed loading assembly to the seed sampling assembly. The seed transport assembly includes multiple retaining members. each retaining member is movable with respect to the seed loading assembly and the seed sampling assembly. The seed transport assembly is operable to position one of multiple retaining members adjacent to the seed loading assembly to engage one of the selected seeds, while positioning another of the retaining members adjacent to the seed sampling assembly to present another of the selected seeds to the seed sampling assembly.
公开号:BR112013001330B1
申请号:R112013001330-3
申请日:2011-07-19
公开日:2019-04-02
发明作者:Kevin L. Deppermann；Michael W. Petersen；Allen N. Ondes；David W. Finley；William M.Fischer；John M.Jensen；David Butruille；Stanton Dotson；Sam Eathington；Heather M.Forbes；Bruce Schnicker；John Tamulonis
申请人:Monsanto Technology Llc；
IPC主号:

专利说明:

Invention Patent Descriptive Report for SEED SAMPLE WITHDRAWAL SYSTEM AND AUTOMATED METHOD FOR REMOVING SEED TISSUE SAMPLES.
CROSS REFERENCE TO RELATED APPLICATION [001] This application claims priority to (and benefit) US Provisional Application No. 61 / 365,826, filed on July 20, 2010, the entire disclosure is incorporated herein by reference.
TECHNICAL FIELD [002] This description relates, in general, to automated systems and methods for removing tissue samples from biological materials, such as seeds, etc.
BACKGROUND [003] This section provides background information related to this description that does not necessarily represent the prior art.
[004] In the development and improvement of plants, genetic improvements are made in the plant, through selective breeding or genetic manipulation, and, when a desirable improvement is achieved, a commercial quantity is developed, or cultivated, when planting and harvesting seeds by several generations. However, not all the seeds harvested express the desired traits and, therefore, these seeds need to be removed from the cultivated quantity. To speed up the process of cultivating the quantity of seeds, statistical samples can be obtained and tested to remove seeds (or groups of seeds associated with the statistical samples) from the seeds that do not adequately express the desired traits. SUMMARY OF THE INVENTION [005] This section provides a general summary of the description and is not a complete description of its full scope or all of its characteristics.
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2/58 [006] In accordance with an aspect of the present description, a seed sampling system includes an automated seed loading assembly operable to select seeds from a plurality of seeds within the seed container, a removal assembly an automated seed sample operable to remove tissue samples from selected seeds and an automated seed transport assembly operable to transfer selected seeds from the seed loading assembly to the seed sample assembly. The seed loading assembly includes multiple retaining members. Each retaining member is movable in relation to the seed loading assembly and the seed sample assembly. The seed transport assembly is operable to position one of the multiple retaining members adjacent to the seed loading assembly to engage one of the selected seeds, while placing another among the retaining members adjacent to the seed sample assembly for present others among the selected seeds at the seed sampling assembly.
[007] According to another aspect of the present description, a seed sampling system includes an automated seed loading assembly that includes a seed container and is operable to separate individual seeds from a plurality of seeds in the seed container , an automated seed transport assembly that includes a transport carousel and multiple retaining member banks mounted on the transport carousel, and an automated seed sample assembly that includes multiple linearly arranged automated extractors adjacent to the transport carousel. Each of the multiple automated extractors is operable to remove a tissue sample from a seed. The transport carousel is configured to rotate around an axis to transport the multiple
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3/58 retaining member banks between extractors and seed loading assembly. The axis of rotation of the transport carousel is substantially parallel to a linear axis defined by the layout of the extractors.
[008] In accordance with yet another aspect of the present description, an automated method for removing tissue samples from seeds is described. The method includes selecting a seed from a plurality of seeds, engaging the selected seed with a retaining member of an automated seed transport assembly, rotating the seed transport assembly around an axis to move the retaining and seed member selected to a position adjacent to an extractor from an automated seed sampling assembly, and remove a tissue sample from the selected seed in the extractor.
[009] Other areas of applicability will become apparent from the description provided in this document. The description and specific examples in the summary are intended to illustrate only and not to limit the scope of this description.
BRIEF DESCRIPTION OF THE DRAWINGS [0010] The drawings described in this document are for illustrative purposes only of the selected modalities and not of all possible implementations, and are not intended to limit the scope of this description.
[0011] Figure 1 is a perspective view of a seed sampling system that includes one or more aspects of the present description and is configured to select seeds and to remove tissue samples from selected seeds;
[0012] Figure 2 is a perspective view of part of a seed loading assembly of the system of Figure 1 illustrating a seed container and a pair of wheels separating the seed loading assembly;
[0013] Figure 3 is a perspective view of part of the mount
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4/58 seed loading gem of the system of Figure 1 illustrating the seed container, the pair of separating wheels, a pair of diverters, and a pair of piping from the seed loading assembly;
[0014] Figure 4 is a perspective view that illustrates the operational relationship between part of the seed loading assembly, a seed transportation assembly, and a seed sample assembly from the system in Figure 1;
[0015] Figure 5A is an elevation view of an exemplary elevator unit of the seed loading assembly of Figure 4 that illustrates a piston of the elevator unit in a neutral position to receive a selected seed from one of the pipes of the seed assembly. seed loading;
[0016] Figure 5B is the elevation view of Figure 5A with the elevator unit piston illustrated in an elevated position to transfer a selected seed to the seed transport assembly;
[0017] Figure 5C is the elevation view of Figure 5A with the elevator unit piston shown in a retracted position to expel a selected seed from the elevator unit;
[0018] Figure 6 is a sectional view of part of the seed loading assembly, the seed transportation assembly and the seed sample removal assembly taken on a plane that includes line 6-6 in Figure 4;
[0019] Figure 7 is a perspective view of an exemplary extractor of the seed sample assembly of Figure 4;
[0020] Figure 8 is an elevation view of an extractor cutting wheel of Figure 7;
[0021] Figure 9 is a schematic illustration placement of a seed selected by a retaining member of the seed transport assembly of Figure 4 adjacent to an extractor from
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5/58 assembly of seed sampling of Figure 4 to remove a tissue sample from the selected seed; and [0022] Figure 10 is an elevation view of another exemplary embodiment of a cutting wheel suitable for use, for example, with the extractor of Figure 7;
[0023] Figure 11 is a perspective view of another exemplary modality of a seed sampling system that includes one or more aspects of the present description and is configured to select seeds and remove tissue samples from selected seeds;
[0024] Figure 12 is an end elevation view of the seed sampling system of Figure 11;
[0025] Figure 13 is a side elevation view of the seed sampling system of Figure 11;
[0026] Figure 14 is a perspective view of an exemplary orientation assembly of the Figure 11 seed sampling system operable to target selected seeds prior to operating the seed sampling system to remove tissue samples from seeds;
[0027] Figure 15 is another perspective view of the orientation assembly of Figure 14; and [0028] Figure 16 is a longitudinal section view of the orientation assembly in Figure 14.
[0029] Corresponding reference numerals indicate corresponding parts throughout all views of the drawings. DETAILED DESCRIPTION [0030] The exemplary modalities will now be more fully described with reference to the attached drawings. Figures 1 to 9 illustrate an exemplary embodiment of an automated seed sampling system 10 that includes one or more aspects of the present description. The illustrated system 10 is suitable for use in removing samples of biological materials. At
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6/58 samples can include, for example, tissue samples, etc. And biological materials can include, for example, seeds (for example, soybeans, corn, wheat, cotton, etc.), etc. The exemplary modality is provided for illustrative purposes only, and can be used in connection with one or more of the methods described in this document.
[0031] As shown in Figure 1, the automated seed sampling system 10 generally includes an automated seed loading assembly 12, an automated seed transport assembly 14 and an automated seed sample assembly 16 Generally, the seed loading assembly 12 operates to select (or isolate, etc.) individual seeds from an amount (e.g., a plurality, etc.) of seeds. The transport assembly 14 then operates to move the selected seeds from the seed loading assembly 12 to the seed sample assembly 16 where the tissue samples are removed from the selected seeds. The tissue samples can be subsequently analyzed to determine whether the seeds, from which the tissue samples were taken, exhibit one or more desired traits or not.
[0032] The operation of the seed loading assembly 12, the seed transportation assembly 14 and the seed sample assembly assembly 16 is automated and can be controlled (and coordinated), for example, by a central control system within the scope of this description. In addition, components of the seed loading assembly 12, the seed transportation assembly 14 and / or the seed sampling assembly 16 can be operated pneumatically using, for example, efficient air flows of about three cubic meters (10 ft ³ ) per minute or less, etc. Such pneumatic operations can be applied to moving seeds through the seed sampling system 10 and between assemblies 12, 14, 16. Such operations
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7/58 pneumatic operations may include taking seeds by the seed sampling system 10 (for example, via vacuum processes, etc.), forcing seeds by system 10 (for example, via air jets, etc.), and / or combinations of these, for example, to help prevent damage to seeds during transport, etc.
[0033] In the illustrated embodiment, the seed loading assembly 12, the seed transportation assembly 14 and the seed sample assembly 16 are supported by various structures such as fixed struts, beams, platforms, pedestals, platforms, etc. . Although such structures are necessary for the construction of the seed sampling system 10, the description of its placement, orientation and interconnections are not necessary for the person skilled in the art to fully and easily understand the structure, function and operation of the sampling system of seed sample 10. In particular, such structures are clearly illustrated by all the Figures and, as such, their locations, orientations and interconnections are easily understood by someone skilled in the art.
[0034] Still with reference to Figure 1, the seed hoppers 18 and 20 are provided to receive seeds in the sampling system 10 as desired and transport the seeds to the seed loading assembly 12. Seed funnels 18 and 20 can be configured (for example, having the size, shape, construction configured, etc.) to receive any type of seed desired (for example, soy, corn, wheat, cotton, etc.) and / or any type of seed quantity within the scope of this description. For example, in the illustrated embodiment, the seed hoppers 18 and 20 can each have the capacity to receive (and taper) about 4,500 soybean seeds for the seed loading assembly 12. Agitators (for example, mechanical mixers, air jets, vibrating devices, etc.) can be provided inside seed hoppers 18 and 20 to promote seed movement
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8/58 tes for mounting seed loading 12 and helping to prevent seeds from forming bridges (or forming empty spaces), adhere, group together, etc. at locations on seed funnels 18, 20.
[0035] Referring now to Figure 2, the seed loading assembly 12 includes a seed container 22 that defines a reservoir 24 for receiving and retaining tapered seeds from seed hoppers 18 and 20 (Figure 1). The seed loading assembly 12 also includes two separation wheels 26 and 28 operably mounted at least partially within the reservoir 24 (and in communication with the seeds in the reservoir 24). The separation wheels 26 and 28 each include openings 30 each communicating with a vacuum source (not shown). The openings 30 (in conjunction with the vacuum source) are configured to capture individual seeds from the seed quantity in reservoir 24 and retain the seeds in the openings 30 as desired. Sensors can be arranged close to each of the separation wheels 26 and 28 to, for example, sense if the individual seeds are captured correctly in the openings 30 (for example, a seed in an opening 30, etc.), to count seeds as they enter the openings 30 (for example, as part of a quality control to monitor the number of seeds entering the seed sampling system 10 and the number of seeds leaving the seed sampling system, etc.), and combinations thereof, etc. And agitators (for example, mechanical mixers, air jets, vibrating devices, etc.) can be provided inside the reservoir 24 to promote the reception of seeds in the openings 30 and help prevent seeds from forming bridges (or forming empty spaces) , join, group, etc. at locations in reservoir 24 where openings 30 receive seeds. In other exemplary embodiments, the sampling system may include seed loading assemblies with more than two separation wheels or less than two separation wheels and / or separation wheels
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9/58 tion with different numbers and / or sizes of openings within them.
[0036] In operation, the separation wheels 26 and 28 rotate (operated by M motors) to move the openings 30 through the reservoir 24 of the seed container 22. In the illustrated embodiment, the separation wheel 26 rotates in a different direction from the wheel separation wheel 28. For example, as can be seen in Figure 2, the separation wheel 26 rotates clockwise and the separation wheel 28 rotates counterclockwise. As each of the separation wheels 26 and 28 rotates, suction is provided to the openings 30 (via the vacuum source) so that the openings 30 that pass through the reservoir 24 capture and retain individual seeds in the openings 30. According to the separation wheels 26 and 28 continue to rotate, they move the openings 30 and capture seeds from the reservoir 24 and to the respective storage compartments 32. In the storage compartments 32, the captured seeds are dislodged from the openings 30 (reduced suction in the openings 30 and / or cleaners (not shown)) and received (for example, via gravity, vacuum, etc.) in a transport chamber (not visible) that extends through one of the corresponding diverters 36 (Figure 3). The separating wheels 26 and 28 then continue to rotate and move the emptied openings 30 back to reservoir 24 over time to capture additional seeds.
[0037] As shown in Figure 3, the diverters 36 of the seed loading assembly 12 are generally arranged below the separation wheels 26 and 28. The diverters 36 are each configured to individually distribute seeds displaced from the separation wheels 26 and 28 for one or two corresponding pipes 38. Derailleurs 36 can operate, for example, to rotate their transport chambers to select positions aligned with one of multiple conduits (not visible) that extend through the corresponding pipes 38 and then transfer ( via gravity, vacuum, mechanical operation, etc.) seeds
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10/58 individual from the transport chamber to the pipelines 38. More particularly, the diverters 36 can each operate to transfer an individual seed from their transfer chamber to one of the pipeline ducts, and then rotate until they line up with a another pipe line and transfer another individual seed to that line. Sensors can be associated with diverters 36 to, for example, sense incoming seeds, count seeds as they enter diverters 36, and combinations thereof, etc. [0038] As shown in Figure 4, multiple elevator units 44 (for example, twelve elevator units 44 in the illustrated embodiment, etc.) of the seed loading assembly 12 are positioned on a bench (or row) (usually below the diverters 36 (Figure 3)) to receive the selected seeds from pipes 38 (Figure 3). Each of the elevator units 44 is in communication with one or more conveying tubes 46 (Figure 3) that extend from a lower portion of each between the pipes 38. The conveying tubes 46 join to inlets 42 defined in a upper block portion 43 of the seed loading assembly with which each of the elevator units 44 is in communication. As such, selected seeds from pipelines 38 can be transferred (for example, via gravity, vacuum, pressurized air, etc.) through transport tubes 46, through upper block portion 43 and up to elevator units 44 for transfer subsequent for the seed transport assembly 14. [0039] With additional reference to Figures 5A to 5C (illustrating an example elevator unit 44), elevator units 44 each include movable pistons 48 (for example, via pneumatic operation, etc.) between a neutral position (Figure 5A), an elevated position (Figure 5B) and a retracted position (Figure 5C). When in the neutral position, each of the pistons 48 can receive seeds from the transport tubes 46 to end portions 50 of the pistons 48. The pistons 48 are then configured to act as
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11/58 seeds from neutral to elevated position for seed transfer / delivery for seed transport assembly 14 (for subsequent transport for seed sample assembly 16). If necessary, the pistons 48 can also be actuated from the neutral position or from the raised position to the retracted position in which the seeds are exposed to an outlet 52 defined in the block portion 43 of the seed loading assembly 12 to expel the seeds (for example, via gravity, vacuum, pressurized air, etc.), as desired, from the elevator units 44 (for example, to a remaining container, another location, etc.). Pistons 48 can be actuated to the stowed position, for example, if deliveries are lost to the seed transport assembly 14, if multiple seeds are detected in one of the elevator units 44 at any given time, if the seeds are detected with one or more specific characteristics (for example, undesirable characteristics, particular sizes, types, particulars, etc. based on intermediate analysis, etc.), etc. Sensors can be associated with elevator units 44 to, for example, sense seeds received from pipes 38, count seeds as they enter diverter elevator units 44, evaluate seeds to be expelled, as desired, from elevator units 44, combinations addition, etc. In addition, the piston end portions 48 may include suction cups (for example, vacuum cups, etc.) for use when receiving and retaining seeds (for example, via negative pressure suction applied to them, for example, through pistons 48, etc.).
[0040] The separation wheels 26 and 28 and diverters 36 of the seed loading assembly 12, in connection with the conduits in the pipes 38, allow the selection of individual seeds from the quantity of seeds that originally passed through the hoppers to the seed container 22. As such, the seed loading assembly 12 operates to supply individual seeds to the
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12/58 seed transport assembly 14 for subsequent transfer to the seed sample assembly assembly 16. Sensors can also be arranged in communication with one or more of the diverters 36 (and their transport chambers), the pipes 38 (and conduits), and / or the transport tubes 46 to help ensure that only one seed at a time is transferred through each transport tube 46 to each elevator unit 44 of the seed loading assembly 12.
[0041] Referring again to Figure 4, the seed transport assembly 14 of the seed sampling system 10 is illustrated positioned adjacent to the elevator units 44 of the seed loading assembly 12. And the sampling assembly of seeds 16 is illustrated positioned adjacent to the seed transport assembly 14. The seed transport assembly 14 includes multiple retaining members 56 mounted on four banks 58a to 58d around a generally tubular transport carousel 60 (for example, four banks 58a to 58d of the twelve retaining members 56 in the illustrated mode, etc.). And the seed sampling assembly 16 includes multiple automated extractors 62 (for example, twelve extractors 62 in the illustrated embodiment, etc.) generally aligned with the retaining members 56. As can be seen, the illustrated embodiment includes the same number elevator units 44, retaining members 56 (on each of the banks 58a to 58d), and extractors 62 so that elevator units 44, retaining members 56 and extractors 62 generally provide sample-taking paths by the seed sampling system 10 for the selected seeds.
[0042] As will be described, the retaining members 56 of the seed transport assembly 14 operate to select (e.g., engage, retain, etc.) the selected seeds from the corresponding elevator units 44 of the seed load assembly 12 and then transfer the seeds to the correct extractors
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13/58 pondentes 62 of the seed sampling assembly 16 for sampling. In the illustrated embodiment, the transport carousel 60 is configured to rotate (for example, via pneumatic operation, electrical operation, etc.) the seats 58a to 58d of the elevator units 44 generally around the rotational axis R (for example, in the counterclockwise as seen in Figure 4, etc.). The rotational axis R of the transport carousel 60 is generally parallel to the linear axis A defined by the multiple automated extractors 62, which are generally arranged linearly. In other exemplary embodiments, the systems may include seed transport assemblies with banks of retaining members configured to rotate differently than is described in this document, for example, around an axis that is substantially perpendicular to a geometric generally defined by automated extractors of these, etc.
[0043] The retaining members 56 of the seed transport assembly 14 are configured to engage and receive seeds from the elevator units 44 (when the pistons 48 of these act seeds to the elevated position) and transport the seeds to the extractors 62 of the system seed sample withdrawal 10. As previously described, the illustrated retaining members 56 are arranged in the four banks 58a to 58d (only three of banks 58a, 58c and 58d are visible in Figure 4). Retaining members 56 are generally evenly distributed along the transport carousel, and seats 58a to 58d are generally evenly distributed around transport carousel 60 (for example, at locations about ninety degrees around the transport carousel) transport 60, etc.). In other exemplary embodiments, the systems may include seed transport assemblies with more or less than four banks of retaining members and / or banks of retaining members oriented differently than is described in this document.
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14/58 [0044] With additional reference to Figure 6, the bench 58a of retaining members 56 is illustrated positioned adjacent to the elevator units 44 for receiving seeds while the bench 58c is positioned adjacent to the extractors 62 of the sample removal assembly seeds 16 to present the seeds to the extractors 62 for sample removal. And banks 58b and 58d are illustrated in idle positions between the seed loading assembly 12 and the seed sample assembly 16. Bank 58b includes a seed shown in transport for the seed sample assembly 16, and bank 58d is empty. After the seeds presented to the extractors 62 by the retention members 56 of the bank 58c are sampled, the retention members 56 release the seeds for subsequent transport as will be described hereinafter. The transport carousel 60 then rotates counterclockwise (as seen in Figure 6) (that is, at an angle of about ninety degrees) to position the bench 58b adjacent to the extractors 62 (to present its seeds to the extractors 62), and bench 58d rotates to a position adjacent to elevator units 44 (to receive additional seeds). Banks 58a and 58c are rotated to idle positions, with each retaining member 56 in bank 58a retaining a seed. The seed transport assembly 14 is operable to continuously rotate the banks 58a to 58d of the retaining members 56 between the elevator units 44 and the automated extractors 62.
[0045] Retaining members 56 include end portions 64 configured to retain seeds received from elevator units 44. In the embodiment illustrated, end portions 64 include suction cups (e.g. vacuum cups, etc.) for use when receiving and retaining the seeds (for example, via negative pressure suction, etc.). The suction cups may include end portions in the shape of a cup, defining, for example, V shapes, U shapes, other shapes, etc. conducive to retain semen
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15/58 tes. The suction cups are configured in such a way that when air pressure is supplied to the suction cups, the seeds can be coupled and retained thereby (with a seed received in a suction cup). In addition, the end portions 64 of the suction cup of the retaining members 56 are movable (for example, via pistons, etc.) relative to the transport carousel 60 (and the seed sampling system 10) to position the seeds in the extractors 62 of the seed sampling system 10. In this way, when the retaining members 56 are rotated to positions adjacent to the extractors 62, the end portions of the suction cup 64 can be actuated (for example, via pistons, etc.) towards the extractors 62 (in relation to the transport carousel 60) to present the seeds for sample removal. In other exemplary embodiments, the systems may include seed transport assembly with retaining members with end portions that define items other than suction cups for use when receiving and retaining seeds, for example, mechanical clamp, gripping mechanisms seed, etc.
[0046] Referring now to Figures 7 and 8, the extractors 62 of the seed sampling assembly 16 each include a cutting wheel 66 operably coupled to a motor 68 (Figure 6) to rotate the cutting wheel 66 during operation. The illustrated cutting wheel 66 includes teeth 70 configured to remove a tissue sample from a seed. The cutting wheel 66 is configured to rotate around an off-center axis 72 during operation. This allows each tooth 70 of the cutting wheel 66 to take a different rotational path in the seed (in relation to the depth) so that each tooth 70 removes a different portion of tissue from the seed, in places that are progressive, growing, etc. deeper into the seed. Figure 10 illustrates another exemplary embodiment of a cutting wheel 66 'that can be used with extractors 62 from the seed sampling assembly 16. In this embodiment, the
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16/58 section 66 'is configured to rotate around a generally central axis 72'. And the cutting wheel 66 'includes teeth 70 each (for example, around 130 of the total teeth 70, etc.) oriented at an angle A (for example, at an angle A of about 57 degrees, etc.). ). In other exemplary embodiments, the systems may include automated extractors with different cutting wheel characteristics to remove tissue samples from seeds, for example, mandrels, lasers, knives, etc.
[0047] With additional reference to the scheme in Figure 9, extractors 62 also each define a channel 74 adjacent to the cutting wheel 66 to guide the movement (for example, orienting, etc.) of seeds to a desired position ( orientation, etc.) inside the extractors 62 (that is, adjacent to the cutting wheel 66). As shown in the example puller 62, channel 74 is defined by two ramp surfaces 76 configured to direct (eg, bypass, etc.) a seed, as needed, in the desired position adjacent to the cutting wheel 66. When the portion end 64 of the retaining member 56 (which holds the seed S) is actuated towards the automated extractor 62, the ramp surfaces 76 guide the seed S in the desired position. The end portion 64 of the retaining member 56 can comprise a flexible material (for example, rubber, etc.) so that the end portion 64 can be actuated (for example, deflected, etc.) as needed to position the seed S at the desired position between the ramp surfaces 76. Sensors can be associated with extractors 62 to, for example, sense incoming seeds, count seeds as they enter extractors 62 from retaining members 56, and combinations thereof, etc. .
[0048] The size and / or shape of the tissue sample removed by the cutting wheels 66 can be adjusted as needed (for example, based on seed size, seed type, sample testing, etc.). For example, the retention system
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17/58 seed sample 10 can adjust the size / shape of a tissue sample by controlling each extractor 62 independently, or alternatively, by controlling any two or more extractors 62 evenly. In addition, extractors 62 can control the position and / or rotation of the cutting wheels 66, based on when the seeds are presented to extractors 62, to ensure incremental removal of tissue samples from the seeds. For example, one or more of the cutting wheels 66 can be attached when the seeds are presented to extractors 62, and then can be subsequently rotated to remove tissue samples from the seeds. Alternatively, one or more of the cutting wheels 66 of the automated extractors 62 may be rotating when the seeds are presented.
[0049] Referring again to Figure 6, after the tissue samples are removed from the seeds, the sampled seeds are captured (for example, released from the retaining members 56 of the seed transport assembly 14 and passed through the funnels, etc. .) and transported through ducts 78 (for example, via gravity, air pressure, air jets, etc.) to a seed tray 80 (Figure 1). And tissue samples are captured (for example, passed through funnels, etc.) and transported through conduits 82 (for example, via gravity, air pressure, air jets, etc.) to a sample tray (not show). The seed tray 80 can include multiple wells (e.g., ninety-six, three hundred and eighty-four, etc.), and the sample tray can include multiple corresponding wells. In addition, the seed tray 80 may include the same number of wells as the sample tray, or a different number (e.g., a multiplicity of these, etc.) in the present description. The seeds are deposited in wells of the seed tray 80, and tissue samples from the seeds are deposited in one or more corresponding wells of the sample tray. As such, the seeds and tissue samples taken
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18/58 of the seeds can subsequently be correlated. Sensors can be positioned (for example, along conduits 78, 82, etc.) to, for example, feel, count, etc. sampled seeds and / or tissue samples received in the sample tray and / or in the seed tray 80.
[0050] The seed and sample trays can be positioned and / or controlled via one or more stages (not shown) movable in XY directions or otherwise to position the trays in relation to the conduits (for example, the conduits 78 and 82, etc.) to ensure that seeds and tissue samples are deposited in the wells of the trays. Additionally, or alternatively, conduits can be structured and / or operable to ensure that tissue samples and seeds are deposited, as desired, without cross-contamination with other tissue / seed samples. In addition, or alternatively, the sample tray can be sealed, via lids, etc., before or shortly after removing the sample tray from the sampling system to limit cross-contamination of the tissue samples retained in each well of the sample tray . The seed tray can be similarly sealed to retain seeds in their respective cavities.
[0051] In the illustrated embodiment, the coupling of seeds by the retaining members 56 in the elevator units 44 and the removal of the seed sample in the automated extractors 62 can occur substantially simultaneously, thereby increasing the performance of the system 10. For example, the yield rate (eg, sampled seed output, etc.) of the illustrated seed sampling system 10 is at least four seeds per second (for example, between about four and six seeds per second , etc.). As such, the seed sampling system 10 can be seen as a high yield system, etc. It should be noted that different numbers of elevator units, of memory
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19/58 holders, and / or extractors as well as different numbers of holding member banks can be provided to adjust the yield rate as desired. In addition, the position of one or more of the components can be modified (for example, the location of the retaining member banks, etc.) to adjust the yield rate of the automated seed sampling system.
[0052] Figures 11 to 16 illustrate another exemplary embodiment of an automated seed sampling system 110 that includes one or more aspects of the present description. The system 110 of this modality is again suitable for use in removing samples of biological materials, and is substantially similar to the system for taking seed samples 10 previously described and illustrated in Figures 1 to 9. In reality, the parts previously described for the seed sampling system 10 can be readily used in connection with the system 110 of this modality, and vice versa.
[0053] As shown in Figures 11 to 13, the seed sampling system 110 generally includes an automated seed loading assembly 112, an automated seed transport assembly 114 and an automated seed sample assembly 116. And the seed loading assembly 112, the seed transportation assembly 114 and the seed sample assembly 116 are substantially similar (and operate in substantially similar ways) to the corresponding assemblies 12, 14 and 16 of the seed system seed sampling 10. As such, the previous descriptions of the seed loading assembly 12, the seed transport assembly 14 and the seed sample assembly 16 apply similarly until now.
[0054] For example, seed hoppers 118 and 120 are provided to receive quantities of seeds in the withdrawal system
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20/58 sample 110 and to direct the seeds to the seed loading assembly 112, which then operates to select (or isolate, etc.) individual seeds received from funnels 118 and 120. The seed loading assembly 112 includes a container seed (not visible) to receive and retain seeds that have passed through the seed hoppers 118 and 120. And the separation wheels (not visible) operate to select individual seeds from the seed container to transport one or more pipes 138 (Figure 13) which, in turn, then distributes the selected seeds to one of the multiple elevator units 144 (for example, twelve elevator units 144 in the illustrated embodiment, etc.) for subsequent transfer to the seed transport assembly 114.
[0055] Also, for example, the seed transport assembly 114 operates to move the selected seeds from the loading assembly 112 to the seed sample removal assembly 116 where tissue samples are removed from the seeds. The seed transport assembly 114 includes multiple retaining members 156 mounted on four benches around a generally tubular transport carousel 160 (e.g., four twelve-member retaining seats 156 in the illustrated embodiment, etc.). The retaining members 156 operate to select (for example, engaging, retaining, etc.) the selected seeds (for example, via suction cups, etc.) from the corresponding elevator units 144 and transfer the seeds (via rotation of the transport 160) for the seed sampling assembly 116. And the seed sampling assembly 116 includes multiple automated extractors 162 (Figure 13) (for example, twelve extractors 162 in the illustrated mode compatible with the number of members of retention 156 in each bank, etc.) generally aligned with the retention members 156 to sample the seeds retained in the retention members 156. The sampled seeds are then received in a 180 seed tray (for example, the seeds
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21/58 are released from retaining members 156 and directed to seed tray 180, etc.), and tissue samples are received in a sample tray (not shown) (for example, tissue samples are directed for the sample tray, etc.).
[0056] In this modality (and as compared to the seed sampling system 10), the seed loading assembly 112 also includes sensors 186 generally positioned below two pipes 138 (Figure 13) and a bank (or row) of multiple guiding units 188 (for example, twelve guiding units 188 in the illustrated embodiment compatible with the number of retaining members 156 in each bank of the seed transport assembly 114 and the number of extractors 162 from the seed sampling assembly 116 , etc.) positioned generally below the two sensors 186 and adjacent to the elevator units 144. The sensors 186 and the orientation units 188 are in communication with multiple transport tubes 146 that extend from a lower portion of each of the pipes 138. The transport pipes 146 extend from the lower portion of the pipes 138, through the sensors 186 and join the inlets 142 (see Figures d and 14 to 16) defined in the upper portions of the guiding units 188. As such, in this modality, selected seeds from the pipes 138 are first transferred by the transport pipes 146 (for example, via gravity, vacuum, pressurized air, etc.) by the sensors 186 and even the orientation units 188 and then transferred from the orientation units 188 to the elevator units 144 for presentation subsequent to the seed transport assembly 114.
[0057] Orientation units 188 are configured to direct selected seeds before transferring seeds to the seed transport assembly 114. As such, also in this modality, the seed transport assembly 114 operates to transport the selected seeds in Particular guidelines
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22/58 res for the 116 seed sample assembly (for example, so that tissue samples can be taken from particular portions of the targeted seeds, so that the seeds can be transferred more safely by the members of retention 156, etc.). In connection with this operation of the seed sampling system 110, the sensors 186 sensors are configured to see if the selected seeds are transferred from pipes 138 to orientation units 188 (for example, as part of a control program for to start operating the guidance units, etc.). Sensors 186 can also (or alternatively) be configured to, for example, count the number of seeds transferred from pipes 138 to orientation units 188, etc. Any suitable sensors can be used for these operations within the scope of this description.
[0058] Figures 14 to 16 illustrate an example of one of the guiding units 188 of the seed loading assembly 112 along with an example of one of the elevator units 144 of the assembly 112. The illustrated guidance unit 188 generally includes a driver 190 configured to target the selected seed and a holder 192 configured to receive the oriented seed from driver 190 in preparation for transfer to elevator unit 144. A transport tube 146 (not shown in Figures 14 to 16) if joins inlet 142 of the guidance unit 188 to transfer a selected seed from one or two of the pipes 138 to the guidance unit 188. The elevator unit 144 is usually located below the support 192. In that position, a piston 148 of the guidance unit elevator 144 (shown in a neutral position in Figures 14 to 16) can engage the oriented seed located in support 192 for subsequent presentation of seed for the seed transport assembly 114 (as previously described, in connection with the seed removal assembly
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23/58 seed sample 10 illustrated in Figures 1 to 9).
[0059] In the illustrated mode, the actuator 190 of the orientation unit 188 moves the received seed to direct the seed. Actuator 190 is mobile (for example, via pneumatic operation, etc.) between a retracted position to receive a selected seed and an extended position to direct the received seed and transfer it to support 192. Sensor 186a is provided to sense when driver 190 is in the stowed position, and sensor 186b is provided to sense when driver 190 is in the extended position. In the stowed position a head portion 190a of the actuator is located in the direction of a recess 194 of the orientation unit 188 so that a selected seed can be received from the entrance 142 and to a guide surface 196 of the orientation unit 188, generally between the head portion 190a and support 192. Moving the actuator 190 from the retracted to the extended position pushes the received seed along the guide surface 196 towards the support 192, causing the seed to roll / bounce to a generally wide surface (for example, a generally flat surface, one of the generally wider surfaces, etc.) of the seed orients along the guide surface 196. Actuator 190 then slides the seed in that orientation (along the generally wide surface of the seed) to the support 192 and generally by the piston 148 of the elevator unit 144 (with the generally wide surface of the seed facing a end portion 148a of piston 148).
[0060] Once the oriented seed is received in the support 192, the support 192 operates to help retain the oriented seed on the piston 148 of the elevator unit 144. For example, the arms 192a and 192b of the support 192 engage so fictional the seed to help retain the seed in its oriented position on the end portion 148a of piston 148. In addition, support 192 can operate to sense (for example, via suitable sensors, etc.) if the seed
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The seed is in a particular and desired orientation (for example, an orientation with the generally wide surface of the seed facing the end portion 148a of piston 148, etc.).
The lift unit 144 then operates the piston 148 from the neutral position to engage and receive the seed at the end portion 148a of the piston 148. In that embodiment, the end portion 148a of the piston 148 includes a vacuum cup for receiving and retain the seed (for example, via negative pressure suction applied to it, for example, through piston 148, etc.) at the end portion 148a of piston 148. Lift unit 144 can then move to an elevated position to transferring / delivering the seed to a retaining member 156 of the seed transport assembly 114 (for subsequent transport to the seed sample assembly 116) or to a retracted position to expel the seed (for example, via gravity, vacuum, pressurized air, etc.) of elevator unit 144 (via outlet 152 between elevator unit 144 and guidance unit 188). For example, as previously described, a desired orientation of the seed may include one in which the generally wide surface of the seed is engaged by the end portion 148a of piston 148. Here, if the seed is in the desired orientation on support 192 (for example , as determined by the perception characteristic of the support 192, etc.), the elevator unit 144 will move the seed oriented to the elevated position and the holding member 156 will engage the oriented seed along a surface in front of the generally wide surface of the seed. Thus, when transferring the seed oriented to the seed sample assembly 116, a tissue sample will be removed from the generally wide seed surface. Alternatively, if the seed is not in the desired orientation, the elevator unit 144 will move the seed to the stowed position and the seed will be expelled from the seed sampling system 110 through outlet 152.
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25/58 [0062] The seed sampling system 110 of this modality can be used to sample any desired type of seed. However, it can be particularly useful to sample wheat seeds that can generally be shaped like a D or generally shaped like a triangle and which typically have wider surfaces (for example, generally flat, etc.) located in front of the embryos of the seeds. As such, guiding units 188 can target wheat seeds prior to sampling (before using extractors 162 to remove tissue samples from wheat seeds) so that tissue samples are removed from wheat seeds in locations away from embryos (for example, locations along the generally wider surfaces of wheat seeds, etc.), for example, to help preserve the germination viability of wheat seeds, etc.
[0063] In some exemplary embodiments, the systems (for example, system 10, system 110, etc.) may include additional assemblies to create images and / or target seeds (for example, in addition to the guidance unit 188, another unit other than guidance unit 188, etc.). For example, seeds can have their image registered and / or be directed before and / or after presentation to sample collection stations. In at least one embodiment, a seed sampling system includes a camera (or other imaging device) to record a seed image, prior to sampling, so that extractors are able to use the size, shape and other features, etc. from the seeds to position the seeds and / or extractors in an appropriate manner (for example, cutting wheels, etc.) so that the desired tissue samples are removed from the seeds. Suitable exemplifying systems and / or methods for registering images and / or targeting seeds are described in U.S. Patent Application Publications 2007/0207485 and 2008/0317279 (the descriptions of which are incorporated herein by way of
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26/58 reference in its entirety).
[0064] For example, the seed sampling system 110 may include an imaging device (for example, a camera, etc.) operable in connection with the guidance unit 188 or, alternatively, operable at the location of the monitoring unit guidance. Here, the imaging device is configured to record the image of a seed, before sampling, so that extractors are able to use the size, shape, other characteristics, etc. of the seeds to position the seeds and / or extractors in an appropriate way (eg cutting wheels, etc.) so that the desired tissue samples are removed from the seeds. This can be used, for example, for seeds in desired positions before sampling, to help analyze seeds after sampling, as part of a quality control program to monitor the operation of the sampling system seed sample 110 (for example, to help adjust (for example, increase speed, decrease speed, etc.)), various processes (for example, seed loading assembly processes 112, seed transportation assembly 114 and the seed sampling assembly 116, etc.) of the seed sampling system 110 during operation and without interrupting the processes, etc.), etc. The various sensors included in the seed sampling system 10 and the seed sampling system 110 can be used in a similar way, for example, to help analyze the seeds after sampling, as part of a program to monitor the operation of seed sampling systems 10, 110, etc.
[0065] In some exemplary embodiments, seed sampling systems may include seed loading assemblies with seed funnel units provided to receive seeds in the sampling systems and for direction
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27/58 the seeds for seed loading assemblies. In these modalities, the seed hopper units can each be configured (for example, having the configured size, the configured shape, the configured construction, etc.) to select seeds (for example, via separation wheels on each among funnel units, etc.) and direct selected seeds to corresponding pipelines (so that each funnel unit processes a separate stream of seeds).
[0066] Seed sampling systems (eg system 10, system 110, etc.) and methods of the present description are operable to protect, preserve, etc. the viability of germination of sampled seeds and can therefore be considered non-destructive. For example, the size, position and / or shape of the removed tissue samples can be precisely controlled to protect the germination viability of the sampled seeds. Germination viability means that a predominant number of sampled seeds (that is, greater than about 50% of all sampled seeds) remains viable after sampling. In a particular modality, at least about 75% of the sampled seeds, and in some modalities at least about 85% of the sampled seeds remains viable. It should be noted that the low rates of germination viability may be tolerable under certain circumstances or for certain applications, for example, as genotyping costs decrease over time because a larger number of seeds could be sampled for less genotyping cost. It should also be noted that sample removal does not need to have any effect on viability.
[0067] In one embodiment, the germination viability of the sampled seeds is maintained for at least about six months after the sample is taken to ensure that the sampled seeds are viable until they reach the field for planting. In a particular modality, the sampled seeds are still treated
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28/58 da to maintain germination viability. Such treatment can generally include any means known in the art to protect a seed from environmental conditions when stored or transported. For example, in one embodiment, the sampled seeds can be treated with a polymer and / or a fungicide to protect the sampled seed when stored or transported to the field before planting.
[0068] The seed sampling systems (for example, system 10, system 110, etc.) of this description can define compact occupied areas. For example, a system (for example, the seed sampling system 10, the seed sampling system 110, etc.) with a seed loading assembly with twelve elevator units, a seed transportation assembly seeds with four banks of twelve holding members, and a seed sampling assembly with twelve extractors can define a footprint of about three meters (10 feet) by about three meters (10 feet), and can have a height of about 2.4 meters (8 feet). Such an occupied area is allowed by the configurations of the seed loading assembly, the seed transportation assembly, and / or the seed sample collection assembly from the system. The compact footprint (and compact size) allows the system to be transported for operation in different locations. Systems with seed loading assemblies with a number of elevator units other than twelve, seed transportation assemblies with a number of banks of the twelve holding members other than four, and seed sampling assemblies with a number of extractors other than twelve can define other occupied areas within the scope of this description.
[0069] The seed sampling systems (eg system 10, system 110, etc.) of this description are configured to accommodate different types of seeds and / or sizes
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29/58 different seeds. For example, separation wheel openings can be configured to accommodate individual seeds of different types and / or sizes (for example, via brushes that automatically adjust for variation in seed sizes, etc.) so that sample withdrawal can be used to process different types of seeds without changing the separation wheels. In addition, the end portions of the retaining members can be configured to retain individual seeds of different types and / or sizes. And extractors can be configured to sample individual seeds of different types and / or sizes.
[0070] Exemplary seeds that can be used with seed sampling systems (eg system 10, system 110, etc.) and methods of the present description include alfalfa seed, apple seed, banana seed , barley seed, bean seed, broccoli seed, cabbage seed, canola seed, carrot seed, castor seed, cauliflower seed, pak choi lettuce seed, citrus seed, clover seed, seed coconut, coffee seed, maize seed, cotton seed, cucumber seed, Douglas fir seed, common bean seed, eggplant seed, eucalyptus seed, fennel seed, common bean seed, zucchini seed round, leek seed, lettuce seed, yellow pine seed, flax seed, melon seed, oat seed, okra seed, olive seed, onion seed, palm seed, pea seed, peanut seed, sement and pepper, poplar seed, pumpkin seed, radiata pine seed, radish seed, rapeseed seed, rice seed, rye seed, spinach seed, sorghum seed, girl pumpkin seed, American pine seed , soybean seed, strawberry seed, sugar beet seed, sugar cane seed, sunflower seed, seed
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30/58 green maize, liquidambar seed, tea seed, tobacco seed, tomato seed, peat seed, watermelon seed, wheat seed and Arabidopsis thaliana seed. And analyzed crops that use the sampled seeds and / or tissue samples obtained as described in this document can forage crops, vegetable oil crops, grain crops, fruit crops, ornamental plants, vegetable crops, fiber crops, crops seasoning, nut crops, peat crops, sugar crops, drink crops, bulb crops, root crops, forest crops, etc.
[0071] In another exemplary embodiment, a seed sampling system includes an automated seed loading assembly operable to select seeds from a plurality of seeds, an automated seed sampling assembly operable to remove samples (e.g. , tissue samples, etc.) of the selected seeds and an automated seed transport assembly operable to transfer the selected seeds from the seed loading assembly to the seed sample assembly. The seed loading assembly includes multiple retaining members, and each of the retaining members is movable in relation to the seed loading assembly and the seed sample assembly. In addition, the seed transport assembly is operable to position one of the multiple retaining members adjacent to the seed loading assembly to engage one of the selected seeds, while placing another among the retaining members adjacent to the seed sample assembly to present others among the selected seeds to the seed sampling assembly.
[0072] In addition, (or alternatively), the seed loading assembly of this modality may include at least one elevator unit operable to actuate the selected seeds in a position to be engaged by one among the multiple members
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31/58 retention of the seed transport assembly. The at least one elevator unit can include a vacuum cup configured to help retain selected seeds in at least one elevator unit. The seed loading assembly can also (or alternatively) include multiple orientation units, each configured to target one of the selected seeds in a desired orientation. The multiple guidance units can each include a trigger configured to target one of the selected seeds and a holder configured to receive the targeted seed from the trigger in preparation for transfer to the seed sampling assembly. The support can also be operable to feel if the seed is in a desired orientation.
[0073] In addition, (or alternatively), the seed transport assembly of this modality may be operable to position one of the multiple retaining members adjacent to the seed loading assembly to engage one of the selected seeds while, substantially at the same time , positions another among the retaining members adjacent to the seed sampling assembly to present another among the selected seeds for the seed sampling assembly. The seed transport assembly may also (or alternatively) include a transport carousel, with each of the multiple retaining members mounted on the transport carousel. And the multiple retaining members can include at least four retaining members arranged circumferentially around the transport carousel. In addition, (or alternatively), each of the multiple retaining members may include a suction cup to retain one of the selected seeds. The suction cup of each of the retaining members can also (or alternatively) be configured to act in relation to the transport carousel to position selected seeds adjacent to the assembly of
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32/58 seed sample.
[0074] In addition, (or alternatively), the seed sampling assembly of this modality may include an extractor to remove a sample from a seed while protecting the germination viability of the sampled seeds. The extractor can be configured to direct the seed to a desired position on the extractor before removing the sample from said seed. For example, the extractor may include a channel configured to guide the seed to a desired position when one of the multiple retaining members acts to position the seed adjacent to the extractor. The channel can be defined by two ramp surfaces configured to direct said seed to the desired position in the extractor. The extractor may include a cutting wheel to remove a sample from a seed. The cutting wheel can be configured to rotate around an off-center axis to thereby remove a progressively deeper sample of the seed as the cutting wheel rotates.
[0075] In addition, (or alternatively), the seed sampling assembly may include multiple extractors aligned, generally, linearly along an axis. And the transport carousel can be operable to rotate around an axis to position one of the retaining members adjacent to at least one of the multiple extractors. Here, the linear axis of the multiple extractors can be directed generally parallel to the rotational axis of the transport carousel.
[0076] The seed sampling system can be operable to process at least about four seeds per second. In addition (or alternatively), the seed extractor system can be operable with any desired type of seed (for example, wheat seeds, corn seeds, cotton seeds, soybean seeds, etc.), and / or can be operable with at least two or more different types of seeds.
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33/58 [0077] In another exemplary embodiment, a seed sampling system includes an automated seed loading assembly with a seed container and it is operable to separate individual seeds from a plurality of seeds inside of the seed container, an automated seed transport assembly that includes a transport carousel and multiple retaining member banks mounted on the transport carousel and an automated seed sample assembly that includes multiple automated extractors arranged linearly along an axis adjacent to the transport carousel, where each of the multiple automated extractors is operable to remove a sample from a seed. The transport carousel is configured to rotate around an axis to transport the multiple banks of retaining members between the extractors and the seed loading assembly. And the axis of rotation of the transport carousel is substantially parallel to the linear axis defined by the layout of the extractors.
[0078] In addition (or alternatively), the seed loading assembly of this modality may include multiple operable elevator units to actuate seeds received from the seed container in a position to be engaged by the seed transport assembly retaining members. The elevator units can each include a vacuum cup configured to help retain seeds in the elevator units. The seed loader system may also (or alternatively) include multiple guidance units configured to target separate seeds in a desired orientation. The multiple guidance units can each include a trigger configured to target one of the selected seeds and a holder configured to receive the driven seed from the trigger in preparation for transfer to the seed sampling assembly. The support can be operable also to feel if the seed is in
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34/58 a desired orientation.
[0079] In addition (or alternatively), the seed transport assembly may be operable to position one of the retaining members adjacent to the seed loading assembly to engage one of the separate seeds, while placing another among the retaining members adjacent to one of the extractors from the seed sampling assembly to present another one of the seeds separated to the extractor. The seed transport assembly may include four banks of substantially uniformly retained members around the transport carousel.
[0080] In addition (or alternatively), the seed sampling assembly of this modality may include several extractors corresponding to several retaining members included in a seed transport assembly bank. For example, the seed sampling assembly can include twelve extractors and each bank in the seed transport assembly can include twelve retaining members. The extractors can be configured to direct the seeds to a desired position on the extractors before removing samples from said seeds. For example, extractors can include channels configured to guide seeds in desired positions when the retaining members act to position the seeds adjacent to the extractors. The channels can be defined by ramp surfaces configured to direct the seeds to the desired positions in the extractors.
[0081] The seed sampling system can be operable to process at least about four seeds per second. In addition (or alternatively), the seed extractor system can be operable with any desired seed type (for example, wheat seeds, corn seeds, cotton seeds, soybean seeds, etc.), and / or can be operable with at least two or more different types of seeds.
[0082] In another example, an automated method
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35/58 used to remove seed samples includes selecting a seed from a plurality of seeds, engaging the selected seed with a retaining member of an automated seed transport assembly, rotating the seed transport assembly around an axis to move the retaining member and the selected seed to a position adjacent to an extractor from an automated seed sampling assembly, and remove a sample from the selected seed in the extractor.
[0083] The coupling of the selected seed to the holding member can occur at about the same time that a sample is removed from another seed selected in the extractor. In addition (or alternatively), engaging the selected seed with a retaining member may include retaining the selected seed on the retaining member using a vacuum.
[0084] In addition, the method may also include (or alternatively) at least one or more of the following operations: act the holding member in the direction of the extractor to present the selected seed to the extractor; receiving the sample removed from the selected seed in a sample tray and receiving the selected seed from which the sample is removed in a seed tray; and direct the selected seed in a desired orientation.
[0085] When the method includes targeting the selected seed in a desired orientation, the operation of removing a sample from the selected seed may include removing a sample from a targeted seed. In addition (or alternatively), the orientation operation may include rolling the selected seed along a surface until the desired portion of the seed is directed along the surface.
[0086] The method can be operable to process at least about four seeds per second. In addition (or alternatively), the method can be operable with any type of seed desired (for example, wheat seeds, corn seeds, semen
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36/58 cotton seeds, soybean seeds, etc.), and / or can be operable with at least one or more of the different types of seeds.
[0087] Seeds and / or tissue samples obtained from the seeds using the seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description can be analyzed as desired. For example, the sampled seeds and / or their tissue samples can be analyzed due to desired traits of interest (for example, physical, chemical, morphological, and / or genetic characteristics; markers; genotypes; etc.), etc. Generally, such traits are determined by analyzing samples for one or more characteristics indicative of at least one genetic or chemical trait. And analyzes can include traces for starch, protein, oil content, determine fatty acid profiles, etc.
[0088] Seeds and / or tissue samples obtained from seeds when using seed sampling systems (for example, system 10, system 110, etc.) and related methods of this description can also be used to facilitate germplasm improvement activities. For example, seeds and / or their tissue samples can be analyzed to identify and select seeds that comprise one or more desired traits, markers and genotypes. In one aspect, analytical methods can be included with the seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description to allow individual seeds that are present in a population batch or volume of seeds are analyzed so that the chemical and / or genetic characteristics of the individual seeds can be determined.
[0089] Non-limiting examples of traits of interest include color (for example, white versus red, etc.), size, shape, type of seed, resistance to pests (for example, insects, mites, fungi, yeasts, molds, bacteria , nematodes, weeds, and
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37/58 parasitic and saprophytic plants, etc.), count of fall number, quality of baking or quality of being turned into paste, etc.
[0090] More particularly, non-limiting examples of characteristics indicative of chemical traits include proteins, oils, carbohydrates, fatty acids, amino acids, biopolymers, pharmaceuticals, starch, fermentable starch, secondary compounds, metabolites, etc. Accordingly, non-limiting examples of chemical traits include amino acid content, protein content, protein composition, starch content, fermentation yield, fermentation efficiency, energy yield, oil content, protein profiling, determination of fatty acid profiles, determination of metabolite profiles, etc.
[0091] And non-limiting examples of characteristics indicative of genetic traits may include, for example, genetic markers, single nucleotide polymorphisms, single sequence repeats, restriction fragment length polymorphisms, haplotypes, SNP tags, genetic marker alleles, genes, DNA-derived sequences, RNA-derived sequences, promoters, untreated 5 'regions of genes, untreated 3' regions of genes, microRNA, siRNA, quantitative trace loci (QTL), satellite markers, transgenes, mRNA, ds mRNA, transcriptional profiles, methylation patterns, etc.
[0092] In one embodiment, seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description can be used to remove tissue samples from wheat seeds. The tissue samples can then be analyzed for any desired characteristics (eg color (eg white versus red, etc.), protein composition, drop number count, baking quality or to be made into paste , etc.). Based on this analysis (for example, based on the presence or absence of one or
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38/58 plus desired characteristics, etc.), sampled wheat seeds can be selected for another use (for example, another analysis, cultivation, packaging, use in breeding operations, etc.).
[0093] In one embodiment, seed samples obtained using seed sampling systems (for example, system 10, system 110, etc.) and related methods include endosperm tissue that allows the determination of frequencies of allele, through which it is possible to infer the parental connection phase for a particular marker. In addition, comparing allele frequency data between two or more germplasm groups provides an understanding of selection targets, whereby alleles that increase in frequency together with a change in the distribution of one or more traits are considered be linked to said feature or said features of interest. In addition, the evaluation of relative allele frequency data between lines can contribute to the construction of genetic link maps.
[0094] In another embodiment, seed samples obtained using seed sampling systems (eg system 10, system 110, etc.) and related methods can be used with folded haploid technologies to contribute with germplasm improvement activities that include the saving of folded haploid programs by selecting only preferred seeds to fold. For example, seed samples can be taken to include folded haploid and haploid material and analyzed for their chemical and genotypic characteristics, and then used in connection with trait integration and trait assessment and marker assisted creation.
[0095] Seeds and / or tissue samples obtained from seeds when using seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description can also be used in a breeding program to select plants or seeds with a genetic trait or
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39/58 desired chemical, in which a genetic trait comprises a genotype, a haplotype, an allele, a sequence, a transcription profile, and a methylation pattern. For example, seeds and / or their tissue samples can be used in combination with any breeding methodology and can be used to select a single generation or to select multiple generations. The choice of the breeding method depends on the plant's reproduction mode, the heritability of the trait (s) to be improved, and the type of cultivar used commercially (for example, hybrid type Fl, cultivar of pure lineage, etc. .). Selected non-limiting approaches to plant creation are presented below. It should also be understood that any commercial and non-commercial cultivars can be used in a breeding program. Factors that include, for example, without limitation, emergency vigor, vegetative vigor, stress tolerance, disease resistance, branching, flowering, seed set, seed size, seed density, sustainability and spotted ability will usually dictate the choice.
[0096] In a particular embodiment, seeds and / or tissue samples obtained from seeds when using seed sampling systems (eg system 10, system 110, etc.) and related methods of this description are used to determine the genetic characteristics of seeds in a marker assisted breeding program. This allows marker-assisted breeding programs in which direct seed sampling (as described in this document) can be conducted while maintaining the identity of individual seeds in the seed sampling systems (for example, system 10, system 110, etc.) to the field. As a result, the marker-assisted breeding program results in a high throughput and a more efficient platform on which a seed population with a desired trait, marker or genotype can be more massive.
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40/58 effectively in less time, with less labor and field resources required. Such advantages will be described more fully below.
[0097] In some exemplary modalities, seeds and / or tissue samples obtained from seeds when using seed sampling systems (eg system 10, system 110, etc.) and related methods of this description can be used in connection with processes to analyze nucleic acids extracted from seeds and / or samples by the presence or absence of at least one genetic marker. Desired seeds can then be selected, based on the results of the nucleic acid analysis, for example, by cultivation plants, etc.
[0098] For example, DNA can be extracted from tissue samples using any DNA extraction methods known to those skilled in the art that will provide sufficient DNA yield, DNA quality, polymerase chain reaction (PCR) response and response to sequencing methods. A non-limiting example of suitable DNA extraction methods is extraction based on sodium dodecyl sulfate (SDS) with centrifugation. In addition, the extracted DNA can be amplified after extraction using any amplification method known to those skilled in the art. For example, a suitable method of application is the preparation of DNA amplification with the GenomiPhi® kit from Amersham Biosciences.
[0099] In addition, (or alternatively), RNA can be extracted from tissue samples using any RNA extraction methods known to those skilled in the art that will provide sufficient RNA yield, RNA quality, PCR response, and response of sequencing methods. A non-limiting example of suitable RNA extraction methods based on SDS extraction with centrifugation with consideration for RNase-free reagents and supplies. In addition, the extracted RNA can be amplified
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41/58 after extraction using any amplification method known to those skilled in the art. For example, a suitable amplification method is Full Spectrum ™ RNA Amplification from System Biosciences.
[00100] The extracted nucleic acids are analyzed for the presence or absence of an appropriate genetic polymorphism. A wide variety of genetic markers for the analysis of genetic polymorphisms are available and are known to those skilled in the art. As used in this document, genetic markers include, but are not limited to, single sequence repeats (SSRs), single nucleotide polymorphisms (SNPs), insertions or deletions (Indels), single characteristic polymorphisms (SFPs) or transcriptional profiles, and nucleic acid sequences. An analysis of nucleic acid by the presence or absence of the genetic marker can be used for seed selection in a breeding population. The analysis can be used to select genes, QTL, alleles, or genomic strains (haplotypes) that comprise or are linked to a genetic marker. In the present document, methods of analysis are known in the art and include, but are not limited to, PCR-based detection methods (e.g., TaqMan tests), microarray methods, and nucleic acid sequencing methods. The genes, alleles, QTL, or haplotypes to be selected can be identified by using newer molecular biology techniques with modifications to classic breeding strategies.
[00101] In one of these exemplifying modalities, sampled seeds are selected based on the presence or absence of one or more characteristics that are genetically linked with a QTL. Examples of QTLs that are often of interest include, but are not limited to, herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, increased grain yield, increased oil, increased nutritional content , increased cultivation rates, tolerance to
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42/58 increased tension, preferred maturity, heightened organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, or traits for improved consumer attraction, or a combination of traits as an index of multiple traits. Alternatively, seeds can be selected based on the presence or absence of one or more characteristics that are genetically linked with a haplotype associated with a QTL. Examples of such QTL that can be included without limitation again are herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, increased grain yield, increased oil, increased nutritional content, increased cultivation, increased stress tolerance, preferred maturity, enhanced organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, or traits for improved consumer attraction, or a combination of traits as a multiple trait index.
[00102] The selection of a breeding population can be started as early as the F2 breeding level, if homozygous inbreeding mother plants are used at the initial breeding crossing. An F1 generation can also be sampled and advanced if one or more of the parent plants of the cross are heterozygous for the alleles or markers of interest. The producer can analyze an F2 population to recover the marker genotype of every individual in the population. Initial population sizes, limited only by the number of seeds available for analysis, can be adjusted to satisfy the desired probability of successfully identifying the desired number of individuals. Accordingly, the probability of finding the desired genotype, the initial population size and the target size of the resulting population can be modified for various breeding methodologies and the level of inbreeding of the sampled population.
[00103] The selected seeds can be swollen or kept
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43/58 taken separately depending on the creation methodology and the target. For example, when a producer is analyzing an F2 population for disease resistance, all individuals with the desired genotype can be swelled and planted in the breeding pond. Conversely, if multiple QTLs with variant effects for a trait such as grain yield are being selected from a given population, the producer can maintain the individual's identity preserved, and go to the field to differentiate individuals with various combinations of the target QTL.
[00104] Various methods of preserving the identity of a single seed can be during the transfer of sampled seeds from the sampling site (for example, from the seed sampling system 10, from the seed sampling system 110 , etc.) to the field. Methods include, however, without limitation, transferring selected individuals (for example, directly from the seed sampling system 10, from the seed sampling system 110, etc.) to trays (for example, the seed tray 80, seed tray 180, etc.), seed strips, a cassette tray, index trays, or transplant the sampled seeds with peat pots, and plant by hand from individual seed packages.
[00105] Multiple selection cycles can be used depending on breeding targets and genetic complexity.
[00106] Advantages of using seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including analytical and seed breeding methods) include, without limitation , reduced labor and required field resources per population or breeding stock, increased ability to assess a larger number of breeding populations per field unit, and increased ability to analyze breeding populations for desired traits prior to planting. Field resources per population are reduced by limiting the space for
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44/58 necessary field to advance the desired genotypes. For example, a population of 1,000 individuals can be planted with 25 seeds per row, with a total of 40 rows being consumed in the field. When using conventional tissue sampling, all 1,000 plants would be labeled and sampled manually when marking the leaf tissue. Results of molecular markers would be necessary before pollination and only those plants that contain the desired genetic composition would be pollinated. Thus, if it were determined that 50 seeds contained in the desired genetic composition, conventional breeding methodology would have required the planting of 1,000 plants to retain the 50 desired seeds. In contrast, the present description allows the producer to analyze the 1,000 seeds in the laboratory and select the desired 50 seeds before planting. The 50 individuals can then be planted in the field, and consume only two rows of 25 seeds. In addition, the present description allows the producer to label and sample in the field, thereby significantly reducing the necessary manual labor resources.
[00107] In addition to reducing the number of field rows per population, use the seed sampling systems (for example, system 10, system 110, etc.) and related methods of this description (including analytical methods and seed breeding) may also allow an increase in the number of populations that the producer can evaluate in a given breeding nursery. Using the example above where 50 seeds from each population of 1,000 seeds contain the desired genetic composition, a producer applying the technology of this description could evaluate 20 populations of 50 seeds in each use the same field area consumed by a single population using conventional tissue sampling techniques in the field. Even if populations are selected by a single allele, use an expected segregation ratio of 1: 2: 1 for an F2 population, the producer can
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45/58 would evaluate 4 populations in the same field area as a single population of tissue sampled in the field.
[00108] Another potential advantage in using seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including analytical and seed breeding methods) is the mitigation of risks associated with plants that are grown in certain geographies where plants may be poorly cultivated or experience poor environmental conditions, or may even be destroyed during storms. For example, seeds with the best genotype or marker composition could be planted in geography 1 and seeds with the almost best genotype could be planted in geography 2. In this case, geography 2 would be a reserve if any problems occurred with the plants grown in geography 1. This is something very difficult to do with the traditional method of taking tissue samples from germinated plants for genotyping, because these plants would then have to be unprotected and transported to the second geography. By using the seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including analytical and seed breeding methods) the problem of transplantation is avoided and also simplifies the logistics of the breeding program.
[00109] In some embodiments, seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including analytical and seed breeding methods) can still be used in a breeding program to introgress a trait in a plant. Here, nucleic acids extracted from tissue samples are analyzed for the presence or absence of at least one genetic marker. Seeds are then selected based on the results of the nucleic acid analysis, and the plants are grown from the selected seeds. Cultivated plants can be used either as
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46/58 parent plants or parent plants in crosses with other plants. [00110] Examples of genetic analysis to select seeds by trait integration include, without limitation, identifying high and recurrent allele frequencies in mother plants, tracking transgenes of interest or mapping the search for the absence of unwanted transgenes, selecting test seed hybrid, select seed that expresses a gene of interest, select seed that expresses an inheritable phenotype, identify seed with selected genetic loci, and zygosity test.
[00111] The identification of high and recurrent allele pair frequencies when using seed sampling systems (eg system 10, system 110, etc.) and related methods of this description (including analytical and seed breeding) allows, again, a reduced number of rows per population and an increased number of populations, or inbred lines, to be planted in a given field unit. Thus, the present description can effectively reduce the resources needed to complete the conversion of inbred lines.
[00112] Seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description and tissue samples obtained therefrom (and the described analytical and seed breeding methods ) also provide quality assurance (QA) and quality control (QC) by ensuring that regulated or unwanted transgenes, unwanted genetic traits, or unwanted inherited phenotypes are identified and discarded before planting. Such an application in a QA capability could effectively eliminate unintended release infractions. Another extension of the present description is to map for the presence of contagious agents and remove contaminated seed before shipment.
[00113] Seed sampling systems (eg system 10, system 110, etc.) and related methods of price
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47/58 this description (and the described analytical and seed breeding methods) can also be applied to identify hybrid seed for transgenic testing. For example, in a conversion from an inbred strain to BCnF stage 1, a producer can effectively create a hybrid seed lot (with the exception of gamete selection) that was 50% hemizygous for the trait of interest and 50% homozygous for the trait to generate hybrid seed for testing. The producer could then analyze all F1 seeds produced at the test crossing and identify and select those seeds that are hemizygous. Such a method is advantageous in that the inferences from the hybrid experiments would represent commercial hybrid genetics with respect to the trace zygosity.
[00114] Other applications of the seed sampling system (for example, system 10, system 110, etc.) and related methods of the present description (including described analytical and seed breeding methods) include use in identifying , track and stack traces of interest, which carry the same advantages identified above with respect to necessary work and field resources. Generally, transgenic conversion programs are run at multi-station locations that carry a much higher cost of administration and land structure. As such, the impact of reducing the need for rows per population or increasing the number of populations in a given field unit is significantly more dramatic on a cost basis than temperate applications.
[00115] Seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including the described analytical and seed breeding methods) can also be used for seeds of plants with two or more transgenes, where accumulating or stacking transgenic regions in plants or strains can be achieved by adding transgenes by transformation, or by crossing parent plants or row
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48/58 genes that contain different transgenic regions, or any combination of these. Analyzes can be conducted to select individual seeds based on the presence of one or more characteristics associated with at least one transgene. Such characteristics include, but are not limited to, a transgene per se, a genetic marker attached to a transgene, mRNA expressed from a transgene, and a protein product of a transgene.
[00116] Furthermore, seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including the described analytical and seed breeding methods) can be used to improve the efficiency of the double haploid program by selecting desired genotypes in the haploid stage and identifying the ploidy level to eliminate non-haploid seeds before they are processed and advance to the field. Both applications again result in the reduction of field resources per population and the ability to assess a large number of populations in a given field unit.
[00117] Folded haploid (DH) plants provide an invaluable tool for plant producers, particularly in the generation of inbreeding strains. A good deal of time is saved when homozygous strains are essentially generated instantly, which eliminates the need for conventional multigenerational inbreeding.
[00118] Particularly, because DH plants are totally homozygous, they are very accessible to quantitative genetics studies. Additive variance and additive x additive genetic variances can be estimated from DH populations. Other applications include identification of epistasis and binding effects. For producers, DH populations were particularly useful in mapping QTL, cytplasmic conversions, and trace introgression. In addition, there is value in testing in evaluating homozygous strains for
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49/58 plant breeding. All genetic variation is between progeny at a breeding crossing, which improves the selection gain.
[00119] However, it is well known in the art that the HD production process is inefficient and can be labor intensive. Although folded haploid plants can occur spontaneously in the wild, this is extremely rare. Most research and breeding applications rely on artificial HD production methods. The initial stage involves the haploidization of the plant which results in the production of a population comprising haploid seed. Non-homozygous strains are crossed with an inducing parent plant, which results in the production of haploid seed. Seed that has a haploid embryo, but a natural triploid endosperm, advances to the second stage. That is, seed and haploid plants are any plant with a haploid embryo, regardless of the ploidy level of the endosperm.
[00120] After selecting haploid seeds from a population, the selected seeds undergo chromosomal duplication to produce folded haploid seeds. Spontaneous chromosomal duplication in a cell line will lead to normal gamete production or the production of unreduced gametes from haploid cell lines. The application of a chemical compound, such as colchicine, can be used to increase the rate of diploidization. Colchicine binds to tubulin and prevents its polymerization in microtubules, thus trapping mitosis in metaphase, it can be used to increase the rate of diploidization, that is, doubling the number of chromosomes. These chimeric plants are self-pollinated to produce diploid seed (folded haploid). This DH seed is cultivated and subsequently evaluated and used in the production of hybrid crossing tests.
[00121] However, processes for producing DH seed generally suffer from low efficacy although methods have been developed in an attempt to increase the frequency of HD production, in
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50/58 including treatment with colchicines. Excellent fabrics include low haploid seed production, reduced gamete viability that results in decreased self-pollination for generation of a DH plant, and inadequate DH seed yield for breeding applications. [00122] Seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including the described analytical and seed breeding methods) represent a breakthrough in seed applications. creation by facilitating the potential for selection in the haploid stage as well as in the diploid seed stage. For example, seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including described analytical and seed breeding methods) can provide sample sampling high yield of an entire haploid seed population, and allow subsequent analysis of samples removed from seeds. This can also provide a high yield crop for an entire population of double haploid seeds. Samples can be analyzed for the presence or absence of one or more characteristics indicative of at least one genetic or chemical trait and, based on the results of the analysis, one or more individual folded haploid seeds can then be selected and plants or plant tissue can be grown from the folded haploid seeds.
[00123] Seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including the analytical and described methods of seed breeding) may also include operations associated with these to analyze seeds for one or more characteristics, such as, for example, genetic markers, transgenes, markers linked to transgenes or diagnoses of these, characteristics related to event performance, event evaluation and trait integration, etc. to determine whether the seeds are in a haploid state or di
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51/58 ploid and / or to select preferred genotypic and phenotypic classes to be duplicated.
[00124] In another embodiment, seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including described analytical and seed breeding methods) can be used with operations to determine the connection phase. When using seed endosperm tissue derived from a diploid plant, the marker haplotypes of the parent plant can be determined by using a genotyping system that allows the detection of different allele frequencies in DNA samples. Since the endosperm tissue is triploid, with two copies derived from the twin gamete, the binding phase of the parent plant lineage can be derived by dissecting heterozygous progeny genotypes (see Figure 1). The DNA sample from the endosperm tissue allows a determination of the ploidy level of the genetic marker. A level of diploid ploidy in the genetic marker indicates maternal inheritance and a level of haploid ploidy in the genetic marker affects paternal inheritance.
[00125] Furthermore, differential allele frequency data can be used to infer the genetic linkage map, but, unlike methods that require haploid material, using the allele frequency described above. Determining the genetic link map is of great use in the context of haplotype characterization, marker (or haplotype) mapping - trait associations. This is particularly robust on a single seed basis, vs. swollen, and is therefore suitable for use in conjunction with seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including the described analytical and breeding methods seed).
[00126] In another embodiment, the seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including the described methods)
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52/58 analytical and seed breeding methods) can also be used in connection with an exam to predict the zygosity of the embryo for a particular gene of interest (GOI). The examination predicts the zygosity of the embryo based on the ratio of the relative copy numbers of a GOI and an internal control gene (IC) per cell or per genome. Generally, this test uses a CI gene that is known zygigosity, for example, homozygous in place (two copies of CI per diploid cell), to normalize the GOI measurement. The ratio of the relative copy numbers from the CI to the GOI predicts the GOI copy number in the cell. In a homozygous cell, for any gene (or single genetic sequence), the number of gene copies is equal to the ploidy level of the cell since the sequence is present in the same place on all homologous chromosomes. When a cell is heterozygous for a particular gene (or hemizygous in the case of a transgene), the gene copy number will be less than the ploidy level of the cell. If the GOI is not detected, the cell is null for the locus, as can happen for a negative segregant from a compromising event or in a mutagenized population. The zygosity of a cell at any locus can thus be determined by the gene copy numbers in the cell.
[00127] In a particular embodiment, seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including the described analytical and seed breeding methods) can be used in connection with an examination to predict zygosity of the corn embryo. In corn seed, the endosperm tissue is triploid, while the embryo tissue is diploid. The endosperm copy number reflects the zygosity of the embryo: a homozygous endosperm (positive or negative) accompanies a homozygous embryo, a heterozygous endosperm (a GOI copy number of 1 or 2) reflects a heterozygous embryo (GOI copy number of 1). Endosperm that is homozygous for the CI will contain three copies of the CI. Copy number of
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53/58
Endosperm GOI can be in the range between 0 (homozygous negative embryo) and 3 (homozygous positive embryo); and copy number of endosperm GOI of 1 or 2 is found in seed where the embryo is heterozygous for GOI (or hemizygous for GOI if GOI is a transgene). The copy number of the endosperm GOI (which can star in the range of 0 to 3 copies) can be determined from the ratio of the copy number of the endosperm IC to the copy number of the endosperm GOI (which can be in the range from 0/3 to 3/3, that is, from 0 to 1), which can then be used to predict the zygosity of the embryo.
[00128] Copy numbers of the GOI or IC can be determined by any examination technique for quantifying copy numbers, as is known in the art. Examples of suitable exams include, but are not limited to, Real Time PCR exams (from TaqMan®) (Applied Biosystems, Foster City, CA) and Invader® (Third Wave Technologies, Madison, WI). Preferably, such tests are developed in such a way that the application efficiency of the IC and GOI sequences are the same or very similar. For example, in a TaqMan® PCR Real-Time PCR exam, the single copy GOI signal (the source cell is determined to be heterozygous for the GOI) will be detected in an amplification cycle after which the signal from an IC two copies, because the quantity of the GOI is half the quantity of the CI. For the same heterozygous sample, an Invader® exam would measure a GOI / IC ratio of about 1: 2 or 0.5. For a sample that is homozygous for GOI and IC, the GOI signal would be detected at the same time as the IC signal (TaqMan®), and the Invader exam would measure a GOI / IC ratio of about 2: 2 or 1.
[00129] These guidelines apply to any polyploid cell, or to haploid cells (such as pollen cells), as the copy number of the GOI or IC remains proportional to the copy number of the genome (or ploidy level) of the cell. Thus, these zygosity tests can be performed on triploid tissues as
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54/58 corn endosperm. In addition, the copy number for a GOI can be measured in addition to 2 copies or in numerically different values than the cell ploidy. The method is still appropriate to detect GOI in polyploids, in some transgenic events with> 2 copies of transgene inserted, after GOI replication by transposition, when GOI exists in autonomously replicating chromosomes or in plasmids and other situations.
[00130] In plant breeding, it is useful to determine zygosity in one or more loci for the purpose of assessing the level of inbreeding (that is, the degree of gene fixation), segregation distortion (that is, in transgenic germplasm, testing for maternal inheritance or for loci that affects gamete fitness), and exogamy level (ie, the relative proportion of homozygosity and heterozygosity). Similarly, the extent of zygosity in one or more loci can be used to estimate hybridity and whether a particular seed lot meets a commercial or regulatory standard for sale as a certified hybrid seed. In addition, in transgenic germplasm, it is useful to know the number of ploidy, or copy number, to distinguish between quality events and to assist in trace integration strategies.
[00131] In another embodiment, seed sampling systems (eg system 10, system 110, etc.) and related methods of the present description (including described analytical and seed breeding methods) can be used in connection with operations to enhance the ability to monitor one or more groups of germplasm for changes in the frequencies of one or more genetic traits, where said genetic traits include markers, alleles, and haplotypes. Methodology is known in the art to compare genetic marker frequency between recently derived populations and their ancestral lines in order to identify those genetic loci that increase in frequency with time team (US Patent ^Nos 5,437,697 and 5,746,023). It is inferred that these loci with frequencies that exceed the allele frequency
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Expected 55/58 were subject to selection. Also, since the predominant selection criterion in breeding programs is income, it is expected that those increasingly frequent alleles can be linked to income.
[00132] In a particular embodiment, seed sampling systems (for example, system 10, system 110, etc.) and related methods of the present description (including the described analytical and seed breeding methods) can be used in connection with operations that allow haplotype-assisted creation. By comparing the frequency of haplotypes in emerging elite lines with the frequency of haplotypes in elite ancestral lines (as determined via pedigree analysis), the identification of haplotypes that deviate from the expected haplotype frequency is possible. In addition, when evaluating estimates of the haplotype effect for said haplotypes, it is also possible to link said haplotypes of increasing frequency with phenotypic results for a set of agronomic traits. The haplotype composition of individual seeds sampled from a plurality of seeds can be determined using genetic markers and seeds with preferred haplotypes are selected and advanced. Thus, more informed breeding decisions and establishment of superior pedigree development programs are allowed by this technology.
[00133] The previous description of the modalities has been provided for the purpose of illustrating and describing. It is not intended to be complete or limiting the invention. Individual elements or characteristics of a particular modality are generally not limited to that particular modality, but, when applicable, are interchangeable and can be used in a selected modality, even if not specifically illustrated or described. It can also be varied in several ways. Such variations should not be considered as a separation from the invention, and it is intended to include all such modifications within the scope of the invention.
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56/58 [00134] The exemplifying modalities were provided so that this description is complete, and fully transmits the scope to those versed in the art. Numerous specific details are presented as examples of specific components, assemblies and methods to provide a complete understanding of the modalities of the present description. It will be apparent to those skilled in the art that specific details do not need to be employed, that exemplifying modalities can be modeled in many different ways and that they should not be interpreted as limiting the scope of the description. In some exemplary embodiments, well-known processes, well-known device structures, and well-known technologies are not described in detail.
[00135] The terminology used in this document was used for the purpose of describing particular exemplifying modalities only and it is not intended to be limiting. As used in this document, the singular forms "one (a)" and "the (a)" may include plural forms as well, unless the context clearly indicates otherwise. The terms understands, understands, includes and has are inclusive and, therefore, specify the presence of characteristics, integers, steps, operations, elements, declared components and / or groups of these, but do not eliminate the presence or addition of one or more among other characteristics, integers, steps, operations, elements, components and / or groups of these. The steps, processes and operations of the method described in this document should not necessarily be interpreted as demanding your achievements in the particular order discussed or illustrated, unless specifically identified as an order of realization. It must also be understood that additional or alternative steps can be employed.
[00136] When an element or layer is referred to as, coupled to, connected to or coupled to another element or layer, it can be directly coupled, connected to or coupled to
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57/58 another element or layer, or interverting elements or layers may be present. On the other hand, when an element is termed as being directly on, directly engaged with, directly connected to or directly coupled to another element or layer, there can be no intervening elements or layers present. Other words used to describe the relationship between elements must be interpreted in a similar way (for example, between versus directly between, adjacent versus directly adjacent, etc.). As used herein, the term and / or includes any and all combinations of one or more within the associated listed terms.
[00137] Although the terms first, second, third, etc. can be used in this document to describe various elements, components, seeds, members and / or sections, those elements, components, seeds, members and / or sections should not be limited by these terms. These terms can be used only to distinguish an element, component, seed, member or section from another element, component, seed, member or section. Terms such as first, second and other numeric terms when used in this document do not imply a sequence or order unless clearly indicated by the context. Thus, a first element, component, seed, member or section discussed below could be referred to as a second element, component, seed, member or section without separating from the teachings of the exemplifying modalities.
[00138] Terms with respect to space, such as internal, external, below, below, lower, above, upper and the like, can be used in this document for an easier description of an element or the relationship of a characteristic with another (s) element (s) or characteristic (s), as illustrated in the Figures. Space-related terms may be intended to cover different orientations of the device in use or in operation in addition to the orientation shown
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58/58 sitting in the figures. For example, if the device in the Figures is turned over, elements described as below or under other elements or characteristics would then be oriented above other elements or characteristics. Thus, the example term below may cover an orientation above or below. The device can be oriented in another way (rotated 90 degrees or in other orientations) and the space descriptors used in this document interpreted accordingly.

权利要求:
Claims (18)
[1]
1. Seed sampling system (10, 110) comprising:
an automated seed loading assembly (12, 112);
an automated seed sample assembly (16, 116); and an automated seed transport assembly (14, 114);
characterized by the fact that:
the sampling assembly (16, 116) has multiple seed extractors (62, 162) aligned along an axis;
the seed transport assembly (14, 114) includes multiple retaining members (56, 156), each of the retaining members (56, 156) being movable in relation to the seed loading assembly (12, 112) and the assembly of seed sampling (16, 116);
the seed transport assembly (14, 114) is operable to rotate the multiple retaining members (56, 156) around an axis so as to position one of the multiple retaining members (56, 156) adjacent to the seed assembly loading of seeds (12, 112) and another among the retaining members (56, 156) adjacent to the seed sampling assembly (16, 116); and the axis around which the seed transport assembly (14, 114) is operable to rotate the multiple retaining members (56, 156) is parallel to the axis along which the multiple extractors (62, 162) are aligned .
[2]
2. System according to claim 1, characterized by the fact that the seed transport assembly (14, 114) includes a rotary transport carousel (60, 160), and in which each of the multiple retaining members ( 56, 156) is mounted on the transport carousel (60, 160).
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2/5
[3]
3. System according to claim 1 or 2, characterized by the fact that each of the multiple retaining members (56, 156) includes a suction cup.
[4]
System according to any one of claims 1 to 3, characterized in that the seed loading assembly (62, 162) includes an elevator unit.
[5]
5. System according to any one of claims 1 to 3, characterized by the fact that each of the multiple seed extractors (62, 162) of the seed sampling assembly (16, 116) includes a channel (74 ) defined by two ramp surfaces (76).
System according to any one of claims 1 to 3, characterized in that the seed loading assembly (62, 162) includes multiple guiding units (188).
[6]
7. System according to claim 6, characterized by the fact that the multiple guiding units (188) each include a driver (190).
[7]
8. Seed sampling system comprising:
an automated seed loading assembly (12, 112) that includes a seed container;
an automated seed transport assembly (14, 114); and an automated seed sampling assembly (16, 116);
characterized by the fact that:
the automated seed transport assembly (14, 114) includes a transport carousel (60, 160) and multiple retaining member banks (56, 156) mounted on the transport carousel (60, 160);
the sample withdrawal assembly (16, 116) automates
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3/5 of it includes multiple linearly automated seed extractors (62, 162) arranged along an axis adjacent to the transport carousel (60, 160); and the transport carousel (60, 160) is configured to rotate around an axis that is parallel to the linear axis defined by the arrangement of automated seed extractors (62, 162).
[8]
9. System according to claim 8, characterized by the fact that the automated seed sample assembly (16, 116) includes several extractors (62, 162) corresponding to several retaining members (56, 156) included in a bank of the seed transport assembly (14, 114).
[9]
10. System according to claim 8, characterized by the fact that each of the extractors (62, 162) includes a channel (74) defined by two ramp surfaces (76).
[10]
11. System according to claim 8 or 9, characterized by the fact that the seed loading assembly (12, 112) includes multiple guiding units (188).
[11]
12. System according to claim 11, characterized by the fact that the multiple guiding units (188) each include an actuator (190).
[12]
13. Automated method for removing tissue samples from seeds, characterized by the fact that the method comprises:
select a seed from a plurality of seeds;
engaging the selected seed with a retaining member (56, 156) of an automated seed transport assembly (14, 114);
rotate the seed transport assembly (14, 114) about an axis oriented parallel to an axis along which the extractors (62,162) of an automated seed sampling assembly (16, 116) are aligned to move the retaining member (56, 156) and the selected seed to an adja position
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4/5 connect to one of the extractors (62, 162) of the automated seed sampling assembly (16, 116); and removing a tissue sample from the selected seed from one of the extractors (62, 162).
[13]
14. Method according to claim 13, characterized in that the engagement of the selected seed with a retaining member (56, 156) includes retaining the selected seed on the retaining member (56, 156) using a vacuum.
[14]
15. Method, according to claim 13, characterized in that it further comprises receiving the tissue sample removed from the selected seed in a sample tray and receiving the selected seed from which the tissue sample was removed in a tray of seed.
[15]
16. Method, according to claim 13, characterized by the fact that it also comprises analyzing the tissue sample for one or more characteristics selected from the group consisting of a genetic marker, a single nucleotide polymorphism, a simple sequence repeat, a restriction fragment length polymorphism, a haplotype, an SNP tag, a genetic marker allele, a gene, a DNA-derived sequence, an RNA-derived sequence, a promoter, an untreated 5 'region of a gene , an untreated 3 'region of a gene, microRNA, siRNA, a QTL, a satellite marker, a transgene, mRNA, ds mRNA, a transcriptional profile and a methylation pattern.
[16]
17. Method, according to claim 13, characterized by the fact that it also comprises analyzing the tissue sample and either selecting or not selecting the seed from which the tissue sample is removed based on the presence of one or more characteristics in the tissue sample that are genetically linked to a QTL selected from the group consisting of herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, grain yield
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5/5 increased, increased oil, increased nutritional content, increased growth rates, increased stress tolerance, preferred maturity, marked organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, traits for improved consumer attraction and a combination of features as an index of multiple features.
[17]
18. Method, according to claim 13, characterized by the fact that it also comprises analyzing the tissue sample and either selecting or not selecting the seed from which the tissue sample was removed based on the presence of one or more characteristics in the sample of tissue that have been genetically linked to a type haplotype associated with a QTL selected from the group consisting of herbicide tolerance, disease resistance, insect or pest resistance, altered fatty acid, protein or carbohydrate metabolism, increased grain yield, oil increased, increased nutritional content, increased growth rates, marked stress tolerance, preferred maturity, marked organoleptic properties, altered morphological characteristics, other agronomic traits, traits for industrial uses, traits for improved consumer attraction and a combination of traits as an index multiple strokes.
[18]
19. Method according to any one of claims 13 to 28, characterized in that it comprises orienting the selected seed in a desired orientation, and in which removing the tissue sample from the selected seed includes removing a tissue sample from the oriented seed .

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同族专利:

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公开号 | 公开日

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CN103118527B|2016-06-08|

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EP2595465B1|2021-06-02|

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BR112013001330A2|2016-05-17|

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AU2011282273A1|2013-01-31|

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EP2595465A4|2018-01-17|

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WO2012012411A3|2012-05-24|

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ZA201300457B|2016-01-27|

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AR082300A1|2012-11-28|

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AU2011282273B2|2015-12-24|

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WO2012012411A2|2012-01-26|

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US20150355205A1|2015-12-10|

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法律状态:
2017-09-12| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2018-05-29| B07A| Technical examination (opinion): publication of technical examination (opinion) [chapter 7.1 patent gazette]|

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2019-01-15| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2019-04-02| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 19/07/2011, OBSERVADAS AS CONDICOES LEGAIS. (CO) 20 (VINTE) ANOS CONTADOS A PARTIR DE 19/07/2011, OBSERVADAS AS CONDICOES LEGAIS |

优先权:

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申请号 | 申请日 | 专利标题

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US36582610P| true| 2010-07-20|2010-07-20|

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US61/365,826|2010-07-20|

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PCT/US2011/044514|WO2012012411A2|2010-07-20|2011-07-19|Automated systems for removing tissue samples from seeds, and related methods|

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