Method of reducing acetylation in plants to improve biofuel production
View Patent ↗The invention provides methods for engineering plants to have reduced levels of acetylation by decreasing expression of one or more Cas1L genes. Such plants can be used, e.g., to increase yield for biofuel production.
1. A method of engineering a plant to reduce acetylation in the stem of a plant; wherein the plant is Arabidopsis , poplar, or rice, the method comprising:
genetically modifying the plant to inhibit expression of all endogenous RWA genes in fiber tissue of the plant, wherein the step of inhibiting expression of each of the endogenous RWA genes in the fiber tissue of the plant comprises introducing one or more expression cassettes into the plant, said one or more expression cassettes encoding one or more polynucleotides that hybridize to the target endogenous RWA nucleic acids, or complements thereof, and inhibit expression of the endogenous RWA genes, expression of which said one or more polynucleotides is driven by a fiber-specific promoter, and
wherein each endogenous RWA gene encodes an RWA polypeptide comprising:
(i) an amino acid sequence YLNRHQTEEWKGWMQVXFLMYHYFXAXEXYNAIRXFIAXYVWMTGFGNFSY YYXXKDFSXXRFXQMMWRLNXXVXXXCXXLXNXYXLYYICPMHTLFTXMVY (SEQ ID NO:12), wherein X at position 17 is L or I; X at position 25 is A or N, X at position 27 is A, T, or V; X at position 29 is Y, I or F; X at position 35 is I, V, L, or M; X at position 39 is A, G, C, or S; X at position 54 is I or V, X at position 55 is R or K; X at position 60 is V, L, or I; X at position 61 is A; X at position 64 is S, A or T; X at position 72 is F or L, X at position 73 is F or L, X at position 75 is A, T, L, or I; X at position 76 is F or V, X at position 77 is C or S; X at position 79 is I or V, X at position 80 is I or V; X at position 82 is N or D; X at position 84 is D or S, X at position 86 is M or T; and X at position 99 is L or V; and
(ii) an amino acid sequence comprising SEQ ID NO:13; and
evaluating acetylation in the stem of the plant and selecting a plant that has reduced acetylation in the stem compared to wild-type.
2. The method of claim 1 , wherein X at position 25 is A, X at position 35 is I, V, or M; X at position 55 is R; X at position 60 is V or L; X at position 75 is A, L, or I; and X at position 86 is M.
3. A plant engineered by the method of claim 1 .
4. Bulk harvested material comprising the plant of claim 3 .
5. Bulk harvested material comprising the plant of claim 3 , wherein the material is present in a fermentation reaction.
6. The method of claim 1 , wherein the plant is a Populus trichcarpa plant.