Feruloyl-CoA:monolignol transferase

Patent metadata

Field	Value
Publication number	US-9441235-B2
Application number	US-201113811823-A
Country	US
Kind code	B2
Filing date	Jul 22, 2011
Priority date	Jul 23, 2010
Publication date	Sep 13, 2016
Grant date	Sep 13, 2016

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.

First claim

What is claimed is:

1. An isolated cDNA nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide, wherein the cDNA comprises SEQ ID NO: 1, or wherein the cDNA encodes a feruloyl-CoA:monolignol transferase polypeptide with SEQ ID NO: 2, or wherein the cDNA encodes a feruloyl-CoA:monolignol transferase polypeptide comprising SEQ ID NO: 2 with conservative amino acid changes to SEQ ID NO: 2.

2. The isolated cDNA 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.

3. The isolated cDNA nucleic acid of claim 1 , wherein the nucleic acid encodes a feruloyl-CoA:monolignol transferase polypeptide with a SEQ ID NO:2 sequence.

4. The isolated cDNA 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 with at least 50% of the activity of a feruloyl-CoA:monolignol transferase with the SEQ ID NO:2.

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

6. A transgenic plant or plant seed comprising the isolated cDNA nucleic acid of any of claim 1 .

7. An expression cassette comprising the feruloyl-CoA:monolignol transferase cDNA nucleic acid of claim 1 operably linked to a promoter functional in a host cell.

8. The expression cassette of claim 7 , wherein the expression cassette is within an expression vector.

9. The expression cassette of claim 7 , wherein the promoter is a promoter functional during plant development or growth.

10. The expression cassette of claim 7 , wherein the promoter is a 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.

11. A plant cell comprising the expression cassette of claim 7 .

12. The plant cell of claim 11 , wherein the plant cell is a monocot cell.

13. The plant cell of claim 11 , wherein the plant cell is a maize, grass or softwood cell.

14. The plant cell of claim 11 , wherein the plant cell is a dicot cell.

15. The plant cell of claim 11 , wherein the plant cell is a hardwood cell.

16. A plant or plant seed comprising the expression cassette of claim 7 .

17. The plant or plant seed of claim 16 , wherein the plant or plant seed is a monocot.

18. The plant or plant seed of claim 16 , wherein the plant or plant seed is a grass, maize or softwood.

19. The plant or plant seed of claim 16 , wherein the plant or plant seed is a gymnosperm.

20. The plant or plant seed of claim 16 , wherein the plant or plant seed is a dicot.

21. The plant or plant seed of claim 16 , wherein the dicot is a hardwood.

22. A method for incorporating monolignol ferulates into lignin of a plant, comprising:
a) stably transforming plant cells with the expression cassette of claim 7 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, to thereby incorporate monolignol ferulates into the lignin of the transgenic plant.

23. The method of claim 22 , wherein the transgenic plant is fertile.

24. The method of claim 22 , further comprising recovering transgenic seeds from the transgenic plant, wherein the transgenic seeds comprise the cDNA nucleic acid encoding a feruloyl-CoA:monolignol transferase.

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

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

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

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

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

30. The method of claim 22 , wherein the lignin in the plant comprises about 1-30% monolignol ferulate.

31. The method of claim 23 , further comprising breeding the 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 a corresponding untransformed plant, and wherein said progeny plant comprises said expression cassette.

32. The method of claim 23 , 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 a corresponding untransformed plant, and wherein said progeny plant comprises said expression cassette.

33. 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.

34. The plant of claim 33 , wherein the plant is a monocot.

35. The plant of claim 33 , wherein the plant is a grass, maize or softwood.

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

37. The plant of claim 33 , wherein the plant is a dicot.

38. The plant of claim 33 , wherein the percent of monolignol ferulates in the plant's lignin is increased relative to the corresponding untransformed plant.

39. The plant of claim 33 , wherein the percent of monolignol ferulates in the plant's lignin is increased by at least 1% relative to the corresponding untransformed plant.

40. The plant of claim 33 , wherein the percent of monolignol ferulates in the plant's lignin is increased by at least 2-5% relative to the corresponding untransformed plant.

41. The plant of claim 33 , wherein the lignin in the plant comprises about 1-30% monolignol ferulates.

42. 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 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.

43. The method of claim 42 , wherein the conditions sufficient to digest the lignin comprise conditions sufficient to cleave ester bonds within monolignol ferulate-containing lignin.

44. The method of claim 42 , wherein the conditions sufficient to digest the lignin comprise mildly alkaline conditions.

45. The method of claim 42 , wherein the conditions sufficient to digest the lignin comprise contact

Assignees

• Wilkerson Curtis
• Ralph John
• Withers Saunia
• Mansfield Shawn D
• Wisconsin Alumni Res Found
• Univ Michigan State
• The Univ Of British Columbia/Univ-Industrial Liaison Office

Inventors

• Wilkerson Curtis
• Ralph John
• Withers Saunia
• Mansfield Shawn D

Classifications

• C12Y203/01133

  Shikimate O-hydroxycinnamoyltransferase (2.3.1.133) · CPC title

• C12P19/04

  Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds · CPC title

• C12N15/8255 Primary

  involving lignin biosynthesis · CPC title

• C08H6/00 Primary

  Macromolecular compounds derived from lignin, e.g. tannins, humic acids · CPC title

• C07G1/00 Primary

  Low-molecular-weight derivatives of lignin (high-molecular-weight derivatives of lignin {C08H6/00}) · CPC title