P-coumaroyl-CoA:monolignol transferase
The invention relates to nucleic acids encoding a p-coumaroyl-CoA:monolignol transferase and to inhibitory nucleic acids adapted to inhibit the expression and/or translation of a p-coumaroyl-CoA:monolignol transferase RNA. Inhibition of p-coumaroyl-CoA:monolignol transferase in plants improves the incorporation of monolignol ferulates into the lignin of plants, giving rise to plant biomass that is more easily processed into useful products such as paper and biofuels.
1. A method of incorporating monolignol ferulates into lignin of a monocot plant comprising:
obtaining a transgenic monocot plant cell comprising an expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a feruloyl-CoA:monolignol transferase polypeptide having at least 95% amino acid sequence identity to SEQ ID NO: 2, SEQ ID NO: 9, SEQ ID NO: 20, or SEQ ID NO: 21, and wherein said transgenic monocot plant cell expresses said feruloyl-CoA:monolignol transferase polypeptide;
reducing or eliminating expression in the transgenic monocot plant cell of at least one endogenous p-coumaroyl-CoA: monolignol transferase gene that encodes a p-coumaroyl-CoA: monolignol polypeptide comprising an amino acid sequence having at least 95% sequence identity to an amino acid sequence as set forth in SEQ ID NO: 30, SEQ ID NO: 35 and SEQ ID NO: 41
by point mutation, a deletion, a missense mutation, an insertion or a nonsense mutation in said endogenous p-coumaroyl-CoA:monolignol transferase gene, or
by expression of an inhibitory nucleic acid comprising a nucleotide sequence having at least 95% nucleotide sequence identity to the nucleotide sequence selected from the group consisting of SEQ ID NO: 48, SEQ ID NO: 53 and SEQ ID NO: 59; and
regenerating one or more of the transgenic monocot plant cells into at least one transgenic monocot plant;
wherein reducing or eliminating expression of the at least one endogenous p-coumaroyl-CoA: monolignol transferase gene incorporates monolignol ferulates into lignin of the transgenic monocot plant.
2. The method of claim 1 , wherein said reducing or eliminating expression of the at least one endogenous p-coumaroyl-CoA: monolignol transferase gene reduces acylation of monolignols with p-coumarate.
3. The method of claim 1 , wherein said reducing or eliminating expression of the at least one endogenous p-coumaroyl-CoA: monolignol transferase gene reduces acylation of monolignols with p-coumarate, and where the monolignols are selected from the group consisting of p-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol.
4. The method of claim 1 , wherein said reducing or eliminating expression of the at least one endogenous p-coumaroyl-CoA: monolignol transferase gene reduces acylation of monolignols with p-coumarate by at least by 30%.
5. The method of claim 1 , wherein the heterologous promoter operably linked to said nucleic acid sequence encoding said feruloyl-CoA:monolignol transferase is selected from the group consisting of a poplar xylem-specific secondary cell wall specific cellulose synthase 8 promoter, cauliflower mosaic virus promoter, a Z10 promoter from a gene encoding a 10 kD zein protein, a Z27 promoter from a gene encoding a 27 kD zein protein, a pea rbcS gene promoter, and an actin promoter from rice.
6. The method of claim 1 , wherein the transgenic monocot plant is a grass species.
7. The method of claim 1 , wherein the transgenic monocot plant is fertile.
8. The method of claim 1 , wherein the method further comprises isolating transgenic monocot plant seeds from the transgenic monocot plant.