FERULOYL-CoA:MONOLIGNOL TRANSFERASE

• US 20130219547A1
• Filed: 07/22/2011
• Published: 08/22/2013
• Est. Priority Date: 07/23/2010
• Status: Active Grant

• Current Assignee: The Board of Trustees of Michigan State University (Michigan State University), The University of British Columbia, Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: The Board of Trustees of Michigan State University (Michigan State University)
• Inventors: Wilkerson, Curtis, Ralph, John, Withers, Saunia
• Granted Patent: US 9,441,235 B2
• Time in Patent Office: Days
• US Class Current: 800/270
• CPC Class Codes: C07G 1/00 Lignin; Lignin derivatives
• CPC Class Codes: C08H 6/00 Macromolecular compounds derived from lignin, e.g. tannins, humic acids
• CPC Class Codes: C12N 15/8255 involving lignin biosynthesis
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 19/04 Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
• CPC Class Codes: C12Y 203/01133 Shikimate O-hydroxycinnamoyltransferase (2.3.1.133)

Abstract

The invention relates to nucleic acids encoding a feruloyl-CoA:monolignol transferase and the feruloyl-CoA:monolignol transferase enzyme that enables incorporation of monolignol ferulates, for example, including p-coumaryl ferulate, coniferyl ferulate, and sinapyl ferulate, into the lignin of plants.

Claims

1. An isolated nucleic acid encoding a feruloyl-CoA:

monolignol transferase wherein the nucleic acid can selectively hybridize to a DNA with a SEQ ID NO;

1 sequence or the nucleic acid has at least about 90% sequence identity with SEQ ID NO;

1.

5. The isolated nucleic acid of claim 1, wherein the nucleic acid encodes a feruloyl-CoA:

monolignol transferase that can catalyze the synthesis of monolignol ferulate(s) from monolignol(s) and feruloyl-CoA.

7. The isolated nucleic acid of claim 1, wherein the nucleic acid encodes a feruloyl-CoA:

monolignol transferase polypeptide with a SEQ ID NO;

2 sequence.

8. The isolated nucleic acid of claim 1, wherein the nucleic acid encodes a feruloyl-CoA:

monolignol transferase that can catalyze the synthesis of monolignol ferulate(s) from a monolignol(s) and feruloyl-CoA with at least about 50%, of the activity of a feruloyl-CoA;

monolignol transferase with the SEQ ID NO;

2.

9. A transgenic plant cell comprising the isolated nucleic acid of claim 1.

10. A transgenic plant comprising the isolated nucleic acid of any of claim 1.

11. An expression cassette comprising the feruloyl-CoA:

monolignol transferase nucleic acid of claim 1 operably linked to a promoter functional in a host cell.

14. The expression cassette of claim 11, wherein the expression cassette is within an expression vector.

15. The expression cassette of claim 11, wherein the promoter is a promoter functional during plant development or growth.

16. The expression cassette of claim 11, wherein the promoter is a poplar xylem-specific secondary cell wall specific cellulose synthase 8 promoter, cauliflower mosaic virus promoter, Z10 promoter from a gene encoding a 10 kD zein protein, Z27 promoter from a gene encoding a 27 kD zein protein, pea rbcS gene or actin promoter from rice.

17. A plant cell comprising the expression cassette of claim 11.

18. The plant cell of claim 17, wherein the plant cell is a monocot cell.

19. The plant cell of claim 17, wherein the plant cell is a maize, grass or softwood cell.

20. The plant cell of claim 17, wherein the plant cell is a dicot cell.

21. The plant cell of claim 17, wherein the plant cell is a hardwood cell.

22. A plant comprising the expression cassette of claim 11.

23. The plant of claim 22, wherein the plant is a monocot.

24. The plant of claim 22, wherein the plant is a grass, maize or softwood.

25. The plant of claim 22, wherein the plant is a gymnosperm.

26. The plant of claim 22, wherein the plant is a dicot.

27. The plant of claim 22, wherein the dicot is a hardwood.

28. A method for incorporating monolignol ferulates into lignin of a plant, comprising:

a) stably transforming plant cells with the expression cassette of claim 11 to generate transformed plant cells;

b) regenerating the transformed plant cells into at least one transgenic plant, wherein feruloyl-CoA;

monolignol transferase is expressed in at least one transgenic plant in an amount sufficient to incorporate monolignol ferulates into the lignin of the transgenic plant.

29. The method of claim 28, wherein the transgenic plant is fertile.

30. The method of claim 28, further comprising recovering transgenic seeds from the transgenic plant, wherein the transgenic seeds comprise the nucleic acid encoding a feruloyl-CoA:

monolignol transferase.

31. The method of claim 28, wherein the plant is a monocot.

32. The method of claim 28, wherein the plant is a grass, maize or softwood plant.

33. The method of claim 28, wherein the plant is a gymnosperm.

34. The method of claim 28, wherein the plant is a dicot.

35. The method of claim 34, wherein the dicot plant is a hardwood.

41. The method of claim 28, wherein the lignin in the plant comprises about 1-30% monolignol ferulate, or about 2-30% monolignol ferulate.

42. The method of claim 29, further comprising breeding the fertile transgenic plant to yield a progeny plant that has an increase in the percentage of monolignol ferulates in the lignin of the progeny plant relative to the corresponding untransformed plant.

43. The method of claim 29, further comprising breeding the fertile transgenic plant to yield a progeny plant that has an increase in the percentage of monolignol ferulates in the lignin of the progeny plant as a dominant trait while still maintaining functional agronomic characteristics relative to the corresponding untransformed plant.

46. A fertile transgenic plant having an increased percent of monolignol ferulates in the plant'"'"'s lignin, the genome of which is stably transformed by the nucleic acid of claim 1, wherein the nucleic acid is operably linked to a promoter functional in a host cell, and wherein the feruloyl-CoA:

monolignol transferase nucleic acid is transmitted through a complete normal sexual cycle of the transgenic plant to the next generation.

47. The plant of claim 46, wherein the plant is a monocot.

48. The plant of claim 46, wherein the plant is a grass, maize or softwood.

49. The plant of claim 46, wherein the plant is a gymnosperm.

50. The plant of claim 46, wherein the plant is a dicot.

51. The plant of claim 46, wherein the percent of monolignol ferulates in the plant'"'"'s lignin is increased relative to the corresponding untransformed plant.

52. The plant of claim 46, wherein the percent of monolignol ferulates in the plant'"'"'s lignin is increased by at least 1% relative to the corresponding untransformed plant.

53. The plant of claim 46, wherein the percent of monolignol ferulates in the plant'"'"'s lignin is increased by at least 2-5% relative to the corresponding untransformed plant.

59. The plant of claim 46, wherein the lignin in the plant comprises about 1-30% monolignol ferulates.

60. A lignin isolated from the transgenic plant comprising the isolated nucleic of claim 1.

61. A method of making a product from a transgenic plant comprising:

(a) providing a transgenic plant that includes an isolated nucleic acid encoding a feruloyl-CoA;

monolignol transferase comprising the isolated nucleic acid of claim 1; and

(b) processing the transgenic plant'"'"'s tissues under conditions sufficient to digest to the lignin; and

thereby generate the product from the transgenic plant,

wherein the transgenic plant'"'"'s tissues comprise lignin having an increased percent of monolignol ferulates relative to a corresponding untransformed plant.

62. The method of claim 61, wherein the conditions sufficient to digest to the lignin comprise conditions sufficient to cleave ester bonds within monolignol ferulate-containing lignin.

63. The method of claim 61, wherein the conditions sufficient to digest to the lignin comprise mildly alkaline conditions.

64. The method of claim 61, wherein the conditions sufficient to digest to the lignin comprise contacting the transgenic plant'"'"'s tissues with ammonia for a time and a temperature sufficient to cleave ester bonds within monolignol ferulate-containing lignin.

65. The method of claim 61, wherein the conditions sufficient to digest to the lignin would not cleave substantially any of the ether and carbon-carbon bonds in lignin from a corresponding plant that does not contain the isolated nucleic acid encoding the feruloyl-CoA:

monolignol transferase.

2. (canceled)

3. (canceled)

4. (canceled)

6. (canceled)

12. (canceled)

13. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)

40. (canceled)

44. (canceled)

45. (canceled)

54. (canceled)

55. (canceled)

56. (canceled)

57. (canceled)

58. (canceled)