The present invention relates to managing operations associated with plant breeding, and more particularly to managing operations associated with seeds on an individual seed basis.
A primary goal of seed companies is to develop seed that grow into plants that are commercially desirable to crop producers. Seed companies devote substantial resources towards research and development of commercially desirable seed.
Conventional research and development techniques tend to be laborious and require vast amounts of land and space. All or much of the seed involved in the research is planted in research plots. After plants emerge from the seed, tissue samples from each plant are acquired. The tissue samples are transported to a laboratory to determine information needed for the research and development of the seed and plants from the seed. These methods are well-known in the industry. The resource costs of land, labor, and machinery are substantial.
Therefore, a need exists in the industry to materially reduce the resources used for evaluating plants and their seed for potential commercial production or further use in plant and seed research and development.
A method for managing seed includes non-destructively sampling individual seeds to acquire evaluation data for each of the individual seeds, storing the evaluation data and seed identifiers associated with each of the individual seeds in a data store, selecting a subset of the sampled seeds for planting at least partially based on the evaluation data, and planting the subset of seeds.
A system for management of seed on an individual seed basis includes an evaluation subsystem for evaluating individual non-destructively sampled seeds to provide evaluation data for the seeds, a selection subsystem for selecting a subset of the individual seeds at least partially based on the evaluation data, a planting subsystem for planting the subset of individual seeds, and a data store for storing seed management data, the seed management data including the evaluation data.
The seed samples of the individual seeds are evaluated to provide genetic information such as phenotype and/or genotype data. Different types of data may be obtained. Of particular interest may be the identification of genetic markers associated with desirable traits. Examples of such traits may include yield traits, disease resistance traits, insect resistance traits, herbicide resistance traits, or tolerance to environmental stresses. In the process of evaluating the seed, any number of forms of analysis may be performed on the tissue sample. The analysis may be genetic, chemical or physical in nature. In addition, the evaluation data may be combined with other data in order to make seed selection decisions.
Where the seed sample is evaluated for genetic data, examples of types of genetic evaluation may include, without limitation, determining if the seed includes a particular genetic marker, determining if the seed includes a particular seed nucleotide polymorphism, determining if the seed includes a particular restriction fragment length polymorphism, a particular haplotype, a particular tag SNP, alleles of a particular genetic marker, a particular gene, a particular DNA-derived sequence, a particular RNA-derived sequence, a particular promoter, a particular siRNA, a particular QTL, a particular transgene or otherwise performing a genetic evaluation.
In step 16, selection occurs. Selection is an important aspect of the plant breeding process. The selection is at least partially based on evaluation data developed during the step of evaluating the seed tissue. Additional data may also be relied upon such as parent information, parent genotype, or other information of potential relevance to plant characteristics of interest. Note that selection occurs prior to the seed being planted. This provides a number of advantages over a process that requires planting and growing of seeds in order to evaluate the seeds, traits, or other characteristics. Efficiencies are achieved because seeds can be excluded without planting them. Thus, fewer resources need be devoted to less promising candidates and more resources can be devoted to more promising candidates. This is an important advantage as growing out seed can use significant resources.
In step 18, the selected individual seeds are stored. The individual seeds may be stored in an indexed system so that the location of any individual seed is known. Thus, the storage system used typically isolates each individual seed from other individual seeds, so that the identity of each individual seed may be maintained. Knowing the identity of each individual seed allows one to store and access data about each individual seed throughout the entire process.
In step 20, the individual seeds may be repositioned. Repositioning of the individual seeds is an optional, but sometimes desirable step. It is contemplated that after seed are stored, it may be desirable to re-index and/or reposition the seed before planting. Doing so may be desirable for several reasons. First, the individual seeds may be re-arrayed into a container which may be directly used by a planter. Second, it is desirable to place the seeds in a particular configuration for planting to gain additional efficiencies. The configuration may include relative positions of the seeds, thus a first seed may be placed next to a second seed where doing so may make the cross pollination process more efficient, or where isolation is necessary.
In step 22, planting of the seeds occurs. Planting of the seed may be performed with an automated single seed planter which is adapted to plant the seeds according to a pre-defined configuration. In addition the automated single seed planter is preferably adapted to collect as-planted data during planting. The as-planted data may include time and position information, such as may be provided by a global positioning system (GPS) receiver. In addition, the as-planted data may include data associated with planting operations. It is further contemplated that exceptions to the pre-defined configuration may occur. Thus, the planter may be further adapted to identify exception conditions which occur during planting.
In step 24, a validation operation is performed. The validation operation may involve a comparison to a pre-determined arrangement for the seeds to as-planted data collected from the planter. It is to be understood, that there may be exceptions to the pre-determined arrangement that occur for any number of reasons. Thus, the validation step allows for the configuration of the seed to be updated based on the as-planted data if necessary.
It should be understood that a data store 48 need not be a physical location, but may be a logical location. A data store may include a logical grouping of data without regard of where it is stored, and which may be stored across multiple locations. Examples of such locations may include, without limitation, one or more database servers, PLC memories, RFID tags, reports, or other locations where data is stored, or any combinations of such locations
A request step is also shown. Data which may be collected at or associated with the request step includes, without limitation, a requestor, a genetic element, an assay or test type, a seed inventory, a request date indicative of the date and/or time a request is made, and a need by time to indicate when the test results are needed.
A sample step 68 is also shown. Data which may be collected at or associated with the sample step 68 may include, without limitation, a sample ID, a seed ID, and a date/time that a sample is taken, assay or test type requested, sample location, and/or seed location. Note that where every seed is individually sampled, the sample ID and the seed ID could be the same.
A lab analysis step 70 is also shown. Data which may be collected at or associated with the lab analysis step 70 may include, without limitation, a sample ID, a seed ID, a date/time of the analysis, results of any assays or tests, and a need by date.
A selection step 76 is shown in
A re-array or re-positioning step 78 is shown in
A planting step 80 is also shown in
A validate step 82 is also shown in
Furthermore, the method is not strictly limited to seeds and would be useful for other propagative structures including meristemic tissue such as sprouted plants, tuber eyes, and the like.
In step 102, a determination is made as to plant characteristics of interest or potential interest. In step 104, corn seed having or potentially having the characteristics of interest is identified. In step 106, the ears of corn are shelled. In step 108, seed chipping (one type of seed sampling) is performed. In step 110, DNA or other types of analysis is performed on the seed chips. In step 112, genotype data is obtained, for example DNA sequences are identified, the presence or absence of particular genetic markers are confirmed and/or or other evaluation data or test results are obtained. In step 114, a seed tray is assembled which includes seeds which have been evaluated. In step 116, a field arrangement which includes seeds of interest is determined. In step 118, the seeds within the seed tray are planted. In step 120, the seeds are harvested. The harvest process in step 120 may use the as-planted data to assist in identifying position of the seed within the field.
A plurality of seeds, for example, corn kernels of different genotypes are analyzed and compared for the purpose of identifying and selecting whether any will be utilized for further research and development or planted to produce commercial or research scale quantities. The method applies as well to other seed specific tests or analyses, as will be apparent to the skilled artisan.
In step 201 candidate seed is identified. One or more factors are used to decide which seed will be a candidate seed for evaluation. In this example, a set of individual candidate seeds, each having a different trait and/or genotype and/or corn variety, are pre-selected. Each candidate seed is isolated from the other candidates but associated with information from which the candidate seeds may be identified in step 202. The identity of each seed may be maintained through the method. Each seed may be identified with an identifier or other code which is stored in the data store 48 or other computer accessible storage. Other methods are possible. In step 203, each candidate seed is assigned an identifying label, such as an identification number.
Pre-selection of candidate seeds can be based on any of a number of factors or criteria. Research scientists may select the factors or criteria used. Examples of types of factors and criteria are commonly known in the art. Some such examples include genotype, phenotype, parentage, traits, or characteristics. Further discussion of these factors or criteria can be found in such references as: (a) Chahal, G. S & Gosal, S. S., 2002. “Principles and Procedures of Plant Breeding”, Alpha Science International, United Kingdom; (b) Falconer, D. S. 1989. “Introduction to Quantitative Genetics”. 3rd Ed. Longman. Burnt Mill; and (c) Frisch, M. & Melchinger, A. E., 2005. “Selection Theory for Marker-assisted Backcrossing.” Genetics 170:909-917; which are incorporated by reference herein.
In step 204 a single candidate seed is isolated by any of a number of ways to present it for removal of specific tissue to gain access to, expose, or sample certain specific tissues(s), part(s), or structure(s) of that seed for testing, or collect the removed tissue for testing. For purposes of this description, tissue(s), part(s), or structure(s) of a seed will collectively sometimes be referred to as tissue. One example of isolation is to place the candidate seed into a cavity or well. Another is to grasp, hold, or restrain the seed by or to some device (e.g. with a vacuum; by clamping action). Another is to apply a substance to the seed which is attracted to or held to a surface or member (e.g. adhesive; magnetic material). Others are possible. The basic function is to hold the seed for accurate and efficient tissue removal and isolate the seed from others, while maintaining identity of the seed.
In step 205, specific tissue is removed from a specified location of the seed. A number of methods can be used. It can be useful, in certain of the methods, to first orient the seed in a certain manner. In some examples, magnetic paint (
An example of tissue removal is with use of a laser. A laser can be precisely controlled in intensity. It also can be focused to a beam width that can be effectively used for removing only a relatively small area of tissue from one side of a seed, and to a relatively small, controlled depth. The laser beam can be operated in a variety of ways to effect tissue removal. An example is programmable raster scanning. The beam is controlled to move at a programmed speed and direction relative to the area to be removed. The laser beam can be focused upon and moved with precision across the seed to ablate the portion of the seed it strikes and remove tissue. Ablation may provide for removing or destroying, cutting, abrading, evaporating (vaporizing), clipping or otherwise. As used herein, ablation refers to such actions, or to analogous actions that remove or separate such seed tissue from the seed. In some instances, this results essentially in a candidate seed having some tissue removed to expose or allow access to internal tissue. The ablation may result in one piece or just a few pieces of removed tissue (more in the sense of cutting or chipping). Alternatively, the ablation may result in the removed tissue being essentially debris (more in the sense of fragments or very small particles, even dust-like, from abrasion, erosive processes, or the like). Alternatively, the ablation may result in the removed material evaporating, sublimating, or forming a plasma. A laser can function in these manners to remove specific tissue from the seed. As mentioned earlier, removed tissue can be collected for testing or analysis. Alternatively, testing or analysis of the remaining seed can be conducted as the tissue removal can be designed to expose or allow access to tissue in the remaining seed. In the case of corn, a laser beam can be controlled to remove an area of the pericarp to gain non-destructive access to underlying seed tissue(s), part(s), or structure(s) of interest, samples of which are collected and used for analysis. In addition a method such as a laser or mechanical slicing may be advantageous over a grinding method, as such methods may reduce potential for contamination between samples.
However, other methods of non-destructive seed tissue removal are possible. One example is a water jet or abrasive jet (e.g. commercially available from Berkeley Chemical Research, Inc., Berkeley, Calif. 94706-026; Flow International Corporation, Kent, Wash. USA; and others). Another is a grinding tool (e.g. Dremel brand MultiPro™ rotary tool) with appropriate sized bit and tip (e.g. engraving, cutting, grinding, carving, sanding, or routing bit tip available at a variety of commercial locations or on-line from Robert Bosch Tool Corporation). Of course, other methods of non-destructive seed tissue removal may be used, and different types may be better suited to different types of seeds, different environments, and other considerations.
In step 206, seed specific analysis is performed on an isolated seed. A number of analyses can be applied to the seed after tissue has been removed, or to the removed tissue from the seed. One example is genetic analysis. By methods known in the art, samples from the seed crown which contains pericarp and endosperm may be assayed for detection of nucleic acids from which genetic information about the seeds can be derived.
An example of one such method is as follows. The ablated seed can be immersed in a polymerase chain reaction (PCR) mixture in preparation for any number of PCR analyses. A detector can generate a signal representative of some aspect of the PCR from which genotyping can be derived. Details of such a signal and its use are well known. A variety of PCR detectors are commercially available. One example is an optical detector for PCR (e.g. Chromo4™ Real-Time PCR Detector from Bio-Rad Laboratories, Inc., Life Science Research Group, 2000 Alfred Nobel Drive, Hercules, Calif. 94547 USA). For example, a sliced portion of the endosperm ma be ground, extracted and amplified via PCR or other amplification process.
In a PCR approach, oligonucleotide primers can be designed for use in PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted from any plant of interest. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed in, inter alia, Innis et al., eds. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press, New York); Innis and Gelfand, eds. (1995) PCR Strategies (Academic Press, new York); and Innis and Gelfand, eds. (1999) PCR Methods Manual (Academic Press, New York), herein incorporated by reference in their entirety. Known methods of PCR include, but are not limited to, methods using paired primers, nested primers, single specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially-mismatched primers, and the like.
In hybridization techniques, all or part of the nucleotide sequence is used as a probe that selectively hybridizes to other corresponding nucleotide sequences present in a population of cloned genomic DNA fragments or cDNA fragments (i.e., genomic or cDNA libraries) from a chosen organism. The hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labeled with a detectable marker. Methods for preparation of probes for hybridization and for construction of genomic libraries are generally known in the art.
Another analysis could be cellular level analysis. An example with respect to corn is described at Gabriella Consonni et al., “Genetic Analysis as a Tool to Investigate the Molecular Mechanisms Underlying Seed Development in Maize”, Annals of Botany 2005 96(3):353-362, which is incorporated by reference herein.
A still further example is nanoscale analysis. See, e.g., Georg H. H. et al., “Analysis of Detergent-Resistant Membranes in Arabidopsis. Evidence for Plasma Membrane Lipid”, Plant Physiol. January 2005; 137(1): 104-116, incorporated by reference herein.
Chemical analysis is another example. A variety of tests can be performed to identify a chemical trait of the tissue, or for other purposes. Other procedures or analyses are, of course, possible. The tissue removal step provides a sample for such analyses. One skilled in the art is familiar with the different analyses and testing that can be done on seed.
In step 207 the results of the analyses from step 206 are recorded. Typically, the results are recorded in an electronic database, but they may be recorded in other forms, such as ink on paper. In step 208 a determination is made as to whether the desired number of candidate seeds have been analyzed. If the desired number of candidate seeds have not yet been analyzed, an additional seed is selected is step 209 and steps 204-208 are repeated on the additional seed. Once the desired number of seeds have been analyzed, the results of the analyses are compared in step 210. Optionally, the analyses can be compared to results from previous assays for the same seed, and/or to data from parental or other non-related seed.
In step 211, a sample is selected based on the comparison of step 210. Once analysis has been completed, results or information from the analysis can be used to, for example, distinguish a seed from other seed, or identify a trait of the seed. This can be used to select one seed over another, or select a seed because of its trait. Of course, any number of other traits may be of interest. Examples of such traits may include herbicide tolerance, disease resistance, insect resistance, pest resistance, nutritional content traits, agronomic traits, industrial use traits, oil content traits, fatty acid content traits, environmental tolerance traits, or other traits of interest. By effective non-destructive sampling of a seed, and by an appropriate genotyping assay, seeds having particular traits (such as those traits indicated by the presence of genetic markers) can be identified.
As diagrammatically illustrated in
Importantly, non-destructive tissue removal and analysis allows such identification to be made without either planting the seed and waiting to test a tissue sample from its growing plant or having to use the land or greenhouse space, labor, and supplies to plant and grow the seed into plants. The controlled, precise, non-destructive removal of seed tissue for testing, or to gain access to relevant underlying tissue or structure for testing, allows analysis to make selections based on tissue of the seed, not on a plant grown from the seed. As can be appreciated, this represents a potential substantial savings in time, labor, and resources, including land resources, for selection processes for seed companies. The controlled, precise non-destructive tissue removal is capable of substantial automation, thus improving through put and efficiency of plant selection processes.
Automated systems may be used for seed handling. An indexed storage system assists in facilitating automatic handling of the seed. The automated system should maintain seeds isolated and separated, such that each seed can be individually identified. The automated seed handling system may be implemented in any number of ways. This may include through vacuum systems, pressurized air systems, mechanical systems, or other types of systems. The re-arraying or re-positioning process (step 20 of
Therefore, methods and systems for single seed management have been described. Numerous variations, options, and alternatives, are contemplated, such as may be apparent to one skilled in the art having the benefit of this disclosure.
This application claims priority under 35 U.S.C. §119 to U.S. Patent Application No. 61/090,961, filed Aug. 22, 2008, hereby incorporated by reference in its entirety.
Number | Date | Country | |
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61090961 | Aug 2008 | US |