Feruloyl-CoA:monolignol transferase

Patent metadata

Field	Value
Publication number	US-10047113-B2
Application number	US-201615237331-A
Country	US
Kind code	B2
Filing date	Aug 15, 2016
Priority date	Jul 23, 2010
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

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:

1. An expression cassette comprising a promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide with at least 95% sequence identity to SEQ ID NO:2.

2. The expression cassette 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 expression cassette of claim 1 , wherein the promoter is a promoter functional during plant development or growth.

4. The expression cassette of claim 1 , 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.

5. The expression cassette of claim 1 , where any amino acid differences in the polypeptide relative to SEQ ID NO:2 are conservative amino acid substitutions.

6. A plant cell comprising an expression cassette comprising a promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide with at least 95% sequence identity to SEQ ID NO:2.

7. The plant cell of claim 6 , wherein the plant cell is a monocot cell, maize cell, grass cell, or softwood cell.

8. The plant cell of claim 6 , wherein the plant cell is a dicot cell, a hardwood cell, or a eucalyptus plant cell.

9. The plant cell of claim 6 , where any amino acid differences in the polypeptide relative to SEQ ID NO:2 are conservative amino acid substitutions.

10. A plant or plant seed comprising an expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide with at least 95% sequence identity to SEQ ID NO:2.

11. The plant or plant seed of claim 10 , wherein the plant or plant seed is a monocot, grass, maize, gymnosperm, or softwood plant or plant seed.

12. The plant or plant seed of claim 10 , wherein the plant or plant seed is a dicot, hardwood, or eucalyptus plant or plant seed.

13. The plant or plant seed of claim 10 , wherein the percent of monolignol ferulates in lignin of the plant or plant grown from the seed is increased at least 1% relative to a corresponding untransformed plant.

14. The plant or plant seed of claim 10 , wherein the percent of monolignol ferulates in the lignin of the plant or plant grown from the seed is increased by at least 2-5% relative to a corresponding untransformed plant.

15. The plant or plant seed of claim 10 , wherein lignin in the plant or a plant grown from the seed comprises about 1-30% monolignol ferulates.

16. The plant or plant seed of claim 10 , wherein the plant is a fertile plant.

17. The plant or plant seed of claim 10 , where any amino acid differences in the polypeptide relative to SEQ ID NO:2 are conservative amino acid substitutions.

18. A product comprising tissue from the plant or plant seed of claim 10 .

19. A method for incorporating monolignol ferulates into lignin of a plant, comprising:
a) stably transforming plant cells with an expression cassette comprising a promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide with at least 95% sequence identity to SEQ ID NO:2 to generate transformed plant cells;
b) regenerating the transformed plant cells into at least one transgenic plant, wherein the feruloyl-CoA:monolignol transferase is expressed in at least one transgenic plant, to thereby incorporate monolignol ferulates into the lignin of the transgenic plant.

20. The method of claim 19 , wherein the transgenic plant is fertile.

21. The method of claim 19 , further comprising recovering transgenic seeds from the transgenic plant, wherein the transgenic seeds comprise the nucleic acid encoding a feruloyl-CoA:monolignol transferase.

22. The method of claim 19 , wherein the plant is a monocot, a grass, a maize, gymnosperm plant, or a softwood plant.

23. The method of claim 19 , wherein the plant is a dicot, hardwood, or eucalyptus plant.

24. The method of claim 19 , wherein the lignin in the plant comprises about 1-30% monolignol ferulate.

25. The method of claim 19 , 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.

26. The method of claim 19 , 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.

27. The method of claim 19 , where any amino acid differences in the polypeptide are conservative amino acid substitutions.

28. A method of making a product from a transgenic plant comprising:
(a) providing a transgenic plant that includes an expression cassette comprising a promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide with at least 95% sequence identity to SEQ ID NO:2; and
(b) processing the transgenic plant's tissues under conditions sufficient to digest the lignin; and
thereby generating 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.

29. The method of claim 28 , wherein the conditions sufficient to digest the lignin comprise conditions sufficient to cleave ester bonds within monolignol ferulate containing lignin.

30. The method of claim 28 , wherein the conditions sufficient to digest the lignin comprise mildly alkaline conditions.

31. The method of claim 28 , wherein the conditions sufficient to digest 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.

32. The method of claim 28 , wherein the conditions sufficient to digest 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.

Assignees

• Wisconsin Alumni Res Foundation Warf
• Univ Michigan State
• Univ British Columbia
• Wisconsin Alumni Res Found

Inventors

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

Classifications

• C12Y203/01133

  Shikimate O-hydroxycinnamoyltransferase (2.3.1.133) · CPC title

• C12N15/8255

  involving lignin biosynthesis · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · 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