Claims
- 1. A method for transforming plant explant tissue, comprising:
a) contacting a cotyledon explant from a plant seedling infected with an Agrobacterium containing DNA to be introduced into the explant with an agent that inhibits enzymatic browning of a wounded plant, plant tissue or plant cell so as to yield transformed explant tissue; and b) identifying transformed explant tissue.
- 2. The method of claim 1 wherein the cotyledon explant is from a legume seedling.
- 3. The method of claim 1 wherein the transformed explant tissue is identified by selection.
- 4. The method of claim 1 further comprising regenerating a differentiated transformed plant from the transformed explant tissue.
- 5. The method of claim 1 wherein the explant is from a dicot.
- 6. The method of claim 1 wherein the explant is from a monocot.
- 7. The method of claim 1 wherein the cotyledon is wounded in the region of the axillary bud or cotyledonary node prior to contacting.
- 8. The method of claim 1 wherein the agent is a sulfhydryl-containing agent.
- 9. The method of claim 1 wherein the agent is cysteine.
- 10. The method of claim 1 wherein the agent is glutathione, sodium thiosulfate, methionine or dithiothreitol.
- 11. The method of claim 1 wherein the agent is an iron chelator.
- 12. The method of claim 1 wherein the agent is a copper chelator.
- 13. The method of claim 1 wherein the agent inhibits plant polyphenol oxidase.
- 14. The method of claim 1 wherein the agent inhibits plant peroxidase.
- 15. The method of claim 1 wherein the agent is present in solid media.
- 16. The method of claim 2 wherein the legume is soybean.
- 17. The method of claim 1 wherein the DNA comprises a selectable gene.
- 18. The method of claim 1 wherein the DNA comprises a detectable gene.
- 19. The method of claim 1 wherein the DNA comprises a promoter operably linked to an open reading frame of interest.
- 20. A plant produced by the method of claim 4.
- 21. A seed produced by the plant of claim 20.
- 22. A method to identify an agent that enhances Agrobacterium-mediated transformation of a plant cell, plant tissue or plant part, comprising:
a) contacting the plant cell, plant tissue or plant part with Agrobacterium containing DNA to be introduced into the plant cell, plant tissue or plant part and the agent so as to yield a transformed plant cell, plant tissue or plant, wherein the agent is not a phenolic compound; and b) detecting or determining whether the agent enhances Agrobacterium-mediated transformation of the plant cell, plant tissue or plant part relative to Agrobacterium-mediated transformation of a plant cell, plant tissue or plant part in the absence of the agent.
- 23. The method of claim 22 wherein the agent is a sulfhydryl-containing agent.
- 24. The method of claim 22 wherein the agent is L-cysteine.
- 25. The method of claim 22 wherein the plant cell, plant tissue or plant part is wounded prior to contacting.
- 26. The method of claim 22 wherein the cell, tissue or part is from a dicot.
- 27. The method of claim 22 wherein the cell, tissue or part is from a monocot.
- 28. The method of claim 22 wherein the cell, tissue or part is from oat.
- 29. The method of claim 22 wherein the cell, tissue or part is from soybean.
- 30. The method of claim 22 wherein the plant cell, tissue or part is regenerable.
- 31. The method of claim 22 wherein the plant tissue is a cotyledon explant.
- 32. The method of claim 22 wherein the agent inhibits plant polyphenol oxidase.
- 33. The method of claim 22 wherein the agent inhibits plant peroxidase.
- 34. The method of claim 22 wherein the agent is an iron chelator.
- 35. The method of claim 22 wherein the agent is a copper chelator.
- 36. An agent identified by the method of claim 22.
- 37. A method for the stable transformation of plant tissue or cells, comprising:
a) contacting plant tissue or cells with an Agrobacterium containing DNA and an agent selected from the group consisting of a sulfhydryl-containing agent, an iron chelator, a copper chelator, an inhibitor of plant polyphenol oxidase and an inhibitor of plant peroxidase; and b) identifying stably transformed plant tissue or cells.
- 38. The method of claim 1 or 37 wherein the efficiency of stable transformation in the presence of the agent is at least 10% greater than the efficiency of transformation in the absence of the agent.
- 39. The method of claim 1 or 37 wherein the efficiency of stable transformation in the presence of the agent is at least 0.5% greater than the efficiency of transformation in the absence of the agent.
- 40. The method of claim 37 wherein the plant tissue or cells are leguminous tissue or cells.
- 41. The method of claim 37 wherein the transformed tissue or cells are identified by selection.
- 42. The method of claim 41 wherein the transformed tissue or cells are selected for in hygromycin.
- 43. The method of claim 3 wherein the transformed tissue is selected for in hygromycin.
- 44. A plant medium comprising: an amount of an agent effective to inhibit the enzymatic browning of a plant organ, tissue or cell, wherein the agent is selected from the group consisting of a sulfhydryl-containing agent, an iron chelator, a copper chelator, an inhibitor of polyphenol oxidase and an inhibitor of peroxidase.
- 45. The medium of claim 44 wherein the agent is cysteine.
- 46. The medium of claim 44 which is aqueous.
- 47. The medium of claim 44 which is a powder.
- 48. The medium of claim 46 wherein the agent is cysteine.
- 49. The medium of claim 48 comprising at least 50 mg/l cysteine.
- 50. The medium of claim 48 comprising at least 100 mg/l cysteine.
- 51. The medium of claim 47 wherein the agent is cysteine.
- 52. The medium of claim 51 which when mixed with a liquid results in a cysteine concentration of at least 50 mg/l.
- 53. The medium of claim 51 which when mixed with a liquid results in a cysteine concentration of at least 100 mg/l.
- 54. The medium of claim 46 wherein the agent is dithiothreiotol.
- 55. The medium of claim 54 comprising 0.75 mM dithiothreiotol.
- 56. The medium of claim 54 comprising 1.25 mM dithiothreiotol.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims the benefit of the filing date of U.S. application Ser. No. 60/170,981, filed Dec. 15, 1999, and U.S. application Ser. No. 60/224,715, filed Aug. 11, 2000, the disclosures of which are incorporated by reference herein.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60170981 |
Dec 1999 |
US |
|
60224715 |
Aug 2000 |
US |