METHOD AND COMPUTER PROGRAM PRODUCT FOR DISTINGUISHING AND SORTING SEEDS CONTAINING A GENETIC ELEMENT OF INTEREST

• US 20090032441A1
• Filed: 04/23/2008
• Published: 02/05/2009
• Est. Priority Date: 04/24/2007
• Status: Active Grant

• Current Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Original Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Inventors: Corak, Steven J., Hunter, James L.
• Granted Patent: US 8,626,337 B2
• Time in Patent Office: Days
• US Class Current: 209/3.300
• CPC Class Codes: A01C 7/044 Pneumatic seed wheels
• CPC Class Codes: B07C 5/3425 of granular material, e.g. ore particles, grain investigation moving fluids or granular solids using optical means G01N21/85
• CPC Class Codes: B07C 5/3427 by changing or intensifying the optical properties prior to scanning, e.g. by inducing fluorescence under UV or x-radiation, subjecting the material to a chemical reaction
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/85 Investigating moving fluids or granular solids

Abstract

A method and computer program product for distinguishing and sort seeds containing a genetic element of interest from a bulk sample. In various embodiments, the present invention comprises associating a marker with at least some of the seeds containing a genetic element of interest of the bulk sample, exciting the seeds using an electromagnetic energy emitting device, evaluating at least some of the seeds of the bulk sample for the presence or absence of the marker using an evaluating device configured to excite a majority of the surface area of the seeds, and sorting the seeds containing a genetic element of interest based on the presence or absence of the marker. In various other embodiments, the method and computer program product may comprise associating a red fluorescent protein marker with at least some of the seeds containing a genetic element of interest of the bulk sample, evaluating at least some of the seeds of the bulk sample for the presence of the red fluorescent protein marker using an evaluating device, and sorting the seeds containing a genetic element of interest based on the presence of the red fluorescent protein marker. In some embodiments, the red fluorescent protein marker is discernable when excited by a certain energy and the evaluating step comprises exciting the seeds containing a genetic element of interest with the certain energy detecting an emission resulting at least in part from the exciting step.

Claims

1. A method for distinguishing seeds containing a genetic element of interest from a bulk sample, the method comprising:

associating a red fluorescent protein marker with at least some of the seeds containing a genetic element of interest of the bulk sample;

evaluating at least some of the seeds of the bulk sample for the presence or absence of the red fluorescent protein marker using an evaluating device; and

sorting the seeds containing a genetic element of interest based on the presence or absence of the red fluorescent protein marker.

2. The method according to claim 1, further comprising:

associating a plurality of supplemental markers with at least some of the seeds containing a corresponding plurality of additional genetic elements of interest of the bulk sample;

evaluating at least one of the seeds of the bulk sample for the presence or absence of the plurality of supplemental markers using the evaluating device; and

sorting the seeds containing the plurality of additional genetic elements of interest based on the presence or absence of the a plurality of supplemental markers.

3. The method according to claim 2, wherein the plurality of supplemental markers are selected from the group consisting of:

yellow fluorescent proteins;

yellow/orange fluorescent proteins;

orange fluorescent proteins;

orange/red fluorescent proteins;

red/orange fluorescent proteins;

red fluorescent proteins;

cyan fluorescent proteins; and

combinations thereof.

4. The method according to claim 1, wherein the sorting step comprises sorting the seeds at a speed of at least about 500 seeds per minute.

5. The method according to claim 1, wherein the sorting step comprises sorting the seeds at a speed of at least about 750 seeds per minute.

6. The method according to claim 1, wherein the red fluorescent protein marker is discernable when excited by a certain energy having a wavelength ranging from substantially about 500 nm to substantially about 580 nm, the certain energy having a peak at substantially about 550 nm.

7. The method according to claim 1, wherein said evaluating step comprises:

exciting the seeds containing a genetic element of interest with the certain energy; and

detecting an emission resulting at least in part from the exciting step, the emission having a wavelength ranging from substantially about 500 nm to substantially about 600 nm, the emission having a peak at substantially about 580 nm.

8. The method of claim 1, wherein the evaluating device comprises a seed color sorter.

9. The method according to claim 1, wherein said evaluating step comprises evaluating seeds for the presence of a marker using at least one image sensing device configured to differentiate between a range of normal seed emissions and an emission from the red fluorescent protein marker.

10. The method according to claim 2, wherein said evaluating step comprises evaluating seeds for the presence of a marker using at least one image sensing device configured to differentiate between a range of normal seed emissions and at least one emission from one or more of the plurality of supplemental markers.

11. The method according to claim 9, wherein the image sensing device is a CCD device or a CMOS device.

12. The method according to claim 9, wherein the evaluating device further comprises at least one filter disposed substantially between the image sensing device and the seeds containing a genetic element of interest, the at least one filter configured for passing the emission from the red fluorescent protein marker to the image sensing device.

13. The method according to claim 10, wherein the evaluating device further comprises at least one tunable filter disposed substantially between the image sensing device and the seeds containing a genetic element of interest, the at least one tunable filter configured for selectively passing the at least one emission from one or more of the plurality of supplemental markers to the image sensing device.

14. The method according to claim 1, wherein the sorting step comprises dispensing the sorted individual seeds into at least one container, the container including only seeds having the red fluorescent protein marker present or only seeds in which the red fluorescent protein marker is absent.

15. The method according to claim 1, further comprising singulating individual seeds from the bulk sample using a singulating device.

16. The method according to claim 15, wherein the singulating step comprises singulating individual seeds using a plurality of elongate hollow structures operatively connected to a vacuum source.

17. The method according to claim 15, wherein the singulating step comprises receiving the bulk sample in a hopper and vibrating the hopper so as to propel individual seeds along an inclined spiral ramp and through a gap configured to permit the passage of one seed at a time.

18. The method according to claim 15, wherein the singulating step comprises:

receiving the bulk sample in a hopper;

feeding seeds from the hopper to a rotatable disk; and

rotating the rotatable disk so as to distribute the seeds to a periphery of the rotatable disk by application of centrifugal force.

19. The method according to claim 15, wherein the singulating step comprises:

receiving the bulk sample in a hopper;

vibrating the bulk sample such that seeds tend to move towards an opening in the hopper;

distributing seeds from the opening in the hopper to a sloped device that includes at least one groove such that a line of individual seeds is formed in the groove; and

depositing individual seeds by gravity force from the sloped device onto a conveying device.

20. A computer program product for distinguishing seeds containing a genetic element of interest that have a red fluorescent protein marker associated with at least some of the seeds containing a genetic element of interest of a bulk sample, the computer program product comprising a computer-readable storage medium having computer-readable program code instructions stored therein comprising:

a first set of instructions for evaluating at least some of the seeds of the bulk sample for the presence or absence of the red fluorescent protein marker using an evaluating device; and

a second set of instruction for sorting the seeds containing a genetic element of interest based on the presence or absence of the red fluorescent protein marker.

21. The computer program product according to claim 20, wherein at least a portion of the seeds of the bulk sample contain a plurality of additional genetic elements of interest associated with a corresponding plurality of supplemental markers, the computer program product further comprising:

a third set of instructions for evaluating at least one of the seeds of the bulk sample for the presence or absence of the plurality of supplemental markers using the evaluating device; and

a fourth set of instructions for sorting the seeds containing the plurality of additional genetic elements of interest based on the presence or absence of the a plurality of supplemental markers.

22. The computer program product according to claim 21, wherein the plurality of supplemental markers are selected from the group consisting of:

yellow fluorescent proteins;

yellow/orange fluorescent proteins;

orange fluorescent proteins;

orange/red fluorescent proteins;

red/orange fluorescent proteins;

red fluorescent proteins;

cyan fluorescent proteins; and

combinations thereof.

23. The method according to claim 20, wherein the second set of computer instructions for sorting comprises sorting the seeds at a speed of at least about 500 seeds per minute.

24. The computer program product according to claim 20, wherein the second set of computer instructions for sorting comprises sorting the seeds at a speed of at least about 750 seeds per minute.

25. The computer program product according to claim 20, wherein the red fluorescent protein marker is discernable when excited by a certain energy having a wavelength ranging from substantially about 500 nm to substantially about 580 nm, the certain energy having a peak at substantially about 550 nm.

26. The computer program product according to claim 25, wherein said first set of instructions comprises:

exciting the seeds containing a genetic element of interest with the certain energy; and

detecting an emission resulting at least in part from the exciting step, the emission having a wavelength ranging from substantially about 500 nm to substantially about 600 nm, the certain energy having a peak at substantially about 580 nm.

27. The computer program product according to claim 20, wherein the evaluating device comprises a seed color sorter.

28. The computer program product according to claim 20, wherein said first set of instructions comprises evaluating seeds for the presence of a marker using at least one image sensing device configured to differentiate between a range of normal seed emissions and an emission from the red fluorescent protein marker.

29. The computer program product according to claim 21, wherein said third set of instructions comprises evaluating seeds for the presence of a marker using at least one image sensing device configured to differentiate between a range of normal seed emissions and at least one emission from one or more of the plurality of supplemental markers.

30. The computer program product according to claim 28, wherein the image sensing device is a CCD device or a CMOS device.

31. The computer program product according to claim 28, wherein the evaluating device further comprises at least one filter disposed substantially between the image sensing device and the seeds containing a genetic element of interest, the at least one filter configured for passing the emission from the red fluorescent protein marker to the image sensing device.

32. The computer program product according to claim 29, wherein the evaluating device further comprises at least one tunable filter disposed substantially between the image sensing device and the seeds containing a genetic element of interest, the at least one tunable filter configured for selectively passing the at least one emission from one or more of the plurality of supplemental markers to the image sensing device.

33. The computer program product according to claim 20, wherein said second set of instructions comprises dispensing the sorted individual seeds into at least one container, the container including only seeds having the red fluorescent protein marker present or only seeds in which the red fluorescent protein marker is absent.

34. The computer program product according to claim 20, further comprising a fifth set of instructions for controlling a singulating device configured for singulating individual seeds from the bulk sample.

35. The computer program product according to claim 34, wherein said fifth set of instructions comprises a set of instructions for singulating individual seeds using a plurality of elongate hollow structures operatively connected to a vacuum source.

36. The computer program product according to claim 34, wherein said fifth set of instructions comprises a set of instructions for receiving the bulk sample in a hopper and vibrating the hopper so as to propel individual seeds along an inclined spiral ramp and through a gap configured to permit the passage of one seed at a time.

37. The computer program product according to claim 34, wherein said fifth set of instructions comprises:

a set of instructions for receiving the bulk sample in a hopper, feeding seeds from the hopper to a rotatable disk, and rotating the rotatable disk so as to distribute the seeds to a periphery of the rotatable disk by application of centrifugal force.

38. The computer program product according to claim 34, wherein said fifth set of instructions comprises:

a set of instructions for receiving the bulk sample in a hopper, vibrating the bulk sample such that seeds tend to move towards an opening in the hopper, distributing seeds from the opening in the hopper to a sloped device that includes at least one groove such that a line of individual seeds is formed in the groove, and depositing individual seeds by gravity force from the sloped device onto a conveying device.

39. A method for distinguishing seeds containing a genetic element of interest from a bulk sample, the method comprising:

associating a marker with at least some of the seeds containing a genetic element of interest of the bulk sample;

exciting the seeds using an electromagnetic energy emitting device;

evaluating at least some of the seeds of the bulk sample for the presence or absence of the marker using an evaluating device; and

sorting the seeds containing a genetic element of interest based on the presence or absence of the marker,

wherein the electromagnetic energy emitting device is configured to excite a majority of the surface area of the seeds.

40. The method according to claim 39, wherein exciting the seeds using an electromagnetic energy emitting device comprises exciting the seeds using at least one illuminating device configured to illuminate a majority of the surface area of the seeds.

41. The method according to claim 40, wherein exciting the seeds using at least one illuminating device further comprises using two or more illuminating devices arranged to collectively illuminate a majority of the surface area of the seeds.

42. The method according to claim 39, wherein evaluating at least some of the seeds of the bulk sample comprises evaluating at least some of the seeds of the bulk sample using at least one image sensing device configured to receive emissions from a majority of the surface area of the seeds.

43. The method according to claim 42, wherein evaluating at least some of the seeds of the bulk sample further comprises using two or more image sensing devices arranged to collectively receive emissions from a majority of the surface area of the seeds.

44. The method according to claim 39, further comprising:

associating one or more supplemental markers with at least some of the seeds containing a corresponding plurality of additional genetic elements of interest of the bulk sample;

evaluating at least one of the seeds of the bulk sample for the presence or absence of the supplemental markers using the evaluating device; and

sorting the seeds containing the plurality of additional genetic elements of interest based on the presence or absence of the supplemental markers.

45. The method according to claim 44, wherein the marker and the supplemental markers are selected from the group consisting of:

green fluorescent proteins;

green/yellow fluorescent proteins;

yellow fluorescent proteins;

yellow/orange fluorescent proteins;

orange fluorescent proteins;

orange/red fluorescent proteins;

red/orange fluorescent proteins;

red fluorescent proteins;

cyan fluorescent proteins; and

combinations thereof.