HIBISCUS CANNABINUS FERULOYL-COA:MONOLIGNOL TRANSFERASE

• US 20150020234A1
• Filed: 10/04/2012
• Published: 01/15/2015
• Est. Priority Date: 10/06/2011
• Status: Active Grant

• Current Assignee: The Board of Trustees of Michigan State University (Michigan State University), 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), Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Wilkerson, Curtis, Ralph, John, Withers, Saunia
• Granted Patent: US 9,493,783 B2
• Time in Patent Office: Days
• US Class Current: 800/278
• CPC Class Codes: C12N 15/8223 Vegetative tissue-specific promoters
• CPC Class Codes: C12N 15/8243 involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
• 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: C12Y 203/01 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12Y 203/01133 Shikimate O-hydroxycinnamoyltransferase (2.3.1.133)

Abstract

The invention relates to isolated nucleic acids encoding a feruloyl-CoA:monolignol transferase and feruloyl-CoA:monolignol transferase enzymes. The isolated nucleic acids and/or the enzymes enable incorporation of monolignol ferulates into the lignin of plants, where such monolignol ferulates include, for example, p-coumaryl ferulate, coniferyl ferulate, and/or sinapyl ferulate. The invention also includes methods and plants that include nucleic acids encoding a feruloyl-CoA:monolignol transferase enzyme and/or feruloyl-CoA:monolignol transferase enzymes.

Claims

1. An isolated Hibiscus cannabinus (Kenaf) feruloyl-CoA:

monolignol transferase nucleic acid with at least about 95% sequence identity to nucleic acid sequence SEQ ID NO;

8.

2. The isolated nucleic acid of claim 1, wherein the nucleic acid encodes a Hibiscus cannabinus (Kenaf) feruloyl-CoA:

monolignol transferase polypeptide with amino acid sequence SEQ ID NO;

9 or SEQ ID NO;

16.

7. An isolated cell comprising the isolated feruloyl-CoA:

monolignol transferase nucleic acid of claim 1.

8. The isolated cell of claim 7, which is a microorganism or a plant cell.

9. A plant comprising the isolated cell of claim 7.

3. An expression cassette comprising a feruloyl-CoA:

monolignol transferase nucleic acid segment with at least about 95% sequence identity to nucleic acid sequence SEQ ID NO;

8, and a promoter operably linked to the nucleic acid segment, wherein the promoter is functional in a host cell.

4. The expression cassette of claim 3, wherein the nucleic acid segment encodes a Hibiscus cannabinus (Kenaf) feruloyl-CoA:

monolignol transferase polypeptide with amino acid sequence SEQ ID NO;

9 or SEQ ID NO;

16.

5. The expression cassette of claim 3, wherein the promoter is a promoter functional or active during plant development or growth.

6. The expression cassette of claim 3, wherein the promoter is a promoter functional or active in woody tissues of a plant.

14. A plant seed comprising the expression cassette of claim 3.

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

(a) stably transforming plant cells with the expression cassette of claim 3, 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.

18. 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 expression cassette of claim 3, wherein the feruloyl-CoA:

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

19. An isolated cell comprising the expression cassette claim 3.

10. A plant comprising a heterologous feruloyl-CoA:

monolignol transferase nucleic acid segment with at least about 95% sequence identity to nucleic acid sequence SEQ ID NO;

8, and a promoter operably linked to the nucleic acid segment, wherein the promoter is functional in a cell of the plant.

11. The plant of claim 10, wherein the plant has an increased percent of monolignol ferulates in the plant'"'"'s lignin.

12. The plant of claim 10, wherein the genome of the plant is stably transformed by the isolated feruloyl-CoA:

monolignol transferase nucleic acid.

13. The plant of claim 10, wherein the feruloyl-CoA:

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

15. A plant seed comprising a heterologous feruloyl-CoA:

monolignol transferase nucleic acid segment with at least about 95% sequence identity to nucleic acid sequence SEQ ID NO;

8, and a promoter operably linked to the nucleic acid segment, wherein the promoter is functional in a cell of the plant.

16. A method for increasing the content of monolignol ferulates in lignin within a plant, comprising:

(a) planting the plant seed of claim 15; and

(b) cultivating a plant germinated from the plant seed, to thereby increase the content of monolignol ferulates in the lignin within the plant.