Methods for simultaneous control of lignin content and composition, and cellulose content in plants
First Claim
1. A method of genetically transforming a plant simultaneously with multiple genes from the phenylpropanoid pathways, comprising incorporating into the genome of the plant a plurality of genes, the genes selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof to produce plants displaying altered agronomic traits.
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Accused Products
Abstract
The present invention relates to a method of concurrently introducing multiple genes into plants and trees is provided. The method includes simultaneous transformation of plants with multiple genes from the phenylpropanoid pathways including 4CL, CAld5H, AldOMT, SAD and CAD genes and combinations thereof to produce various lines of transgenic plants displaying altered agronomic traits. The agronomic traits of the plants are regulated by the orientation of the specific genes and the selected gene combinations, which are incorporated into the plant genome.
55 Citations
72 Claims
- 1. A method of genetically transforming a plant simultaneously with multiple genes from the phenylpropanoid pathways, comprising incorporating into the genome of the plant a plurality of genes, the genes selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof to produce plants displaying altered agronomic traits.
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16. A method of preparing plant cells having in their genome a plurality of DNA constructs, the method comprising
a) incorporating into the genome of the cells a plurality of DNA constructs to yield transformed cells, each construct comprising a polynucleotide sequence encoding a protein selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof, operably linked to a promoter sequence functional in the cells, and a termination sequence b) identifying the transformed plant cells, the genome of which is augmented with DNA from the different DNA constructs.
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24. A method of preparing transgenic plants having altered lignin and cellulose compositions, the method comprising
a) providing the genome of the plants with a plurality of DNA constructs to yield transformed plant cells, each construct comprising a polynucleotide sequence encoding a protein selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof, the polynucleotide sequence operably linked to a promoter sequence functional in the plant cells, and a termination sequence; -
b) regenerating the transformed plant cells to yield transgenic plants, the genome of which is augmented with DNA from different DNA constructs, and c) expressing the DNA constructs in the cells of the transgenic plants in an amount effective to alter the lignin and cellulose composition in the plants.
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35. A method of preparing a transgenic tree comprising
a) incorporating into the genome of the tree a plurality of desired DNA constructs to produce transformed tree cells, each construct comprising a polynucleotide sequence encoding a protein selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof, operably linked to a promoter sequence functional in the cells, and a termination sequence; -
b) regenerating the transformed tree cells to yield transgenic trees, the genome of which is augmented with the plurality of DNA constructs; and
c) expressing the DNA construct in the cells of the transgenic tree in an amount effective to alter the lignin and cellulose composition of the tree.
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38. A plant having incorporated into its genome a DNA construct comprising a polynucleotide sequence encoding a protein selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof, operably linked to a promoter sequence, and a termination sequence.
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57. A plurality of DNA constructs, each construct comprising in the 5′
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direction;
a) a gene promoter sequence, b) a gene termination sequence; and
c) a polynucleotide sequence encoding a protein selected from the group consisting of 4CL, CAld5H, AldOMT, CAD, and SAD, substantially similar fragments thereof, and combinations thereof involved in a lignin biosynthetic pathway, the polynucleotide sequence operably linked to the promoter and termination sequences.
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61. A plurality of DNA constructs incorporated into the genome of gymnosperms, the constructs comprising nucleotide sequences encoding CAld5H, AldOMT, and SAD genes, a substantially similar fragments thereof and combinations thereof, in sense orientation, to produce syringyl lignin as compared to a non-transformed plant.
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62. A plurality of DNA constructs incorporated into the genome of gymnosperm, the constructs comprising 4CL gene or substantially similar fragments thereof in sense or antisense orientation and CAld5H, AldOMT, and SAD genes, substantially similar fragments thereof and combination thereof in sense orientation to decrease lignin content, increase syringyl/guaiacyl (S/G) lignin ratio and increase cellulose content as compared to a non-transformed plant.
- 63. A plurality of DNA constructs incorporated into the genome of gymnosperms, the constructs comprising 4CL and CAD genes or substantially similar fragments thereof in sense or antisense orientation and CAld5H, AldOMT, and SAD genes, substantially similar fragments thereof and combination thereof in sense orientation to decrease lignin content, increase S/G lignin ratio and increase cellulose content as compared to a non-transformed plant.
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64. A DNA construct incorporated into the genome of plants, comprising 4CL gene or substantially similar fragments thereof in the sense orientation to increase lignin content compared to a non-transformed plant.
- 66. A plurality of DNA constructs incorporated into the genome of plants, the constructs comprising 4CL, CAld5H, AldOMT, and SAD genes, substantially similar fragments thereof or combination thereof, in the sense orientation, to increase lignin content and increase S/G ratio compared to a non-transformed plant.
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68. A plurality of DNA constructs incorporated into the genome of plants, the constructs comprising 4CL, CAld5H, AldOMT, and SAD genes, substantially similar fragments thereof and combination thereof in the sense orientation and CAD gene or substantially similar fragments thereof in the antisense orientation to increase lignin content and increase S/G ratio compared to a non-transformed plant.
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70. A plurality of DNA constructs incorporated into the genome of angiosperms, the constructs comprising CAld5H, AldOMT, and SAD genes, substantially similar fragments thereof and combinations thereof in sense orientation to engineer high S/G lignin ratio compared to a non-transformed plant.
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71. A plurality of DNA constructs incorporated incorporating into the genome of angiosperms, the constructs comprising 4CL gene or substantially similar fragment thereof in sense or antisense orientation and CAld5H, AldOMT, and SAD genes, substantially similar fragment thereof, and combinations thereof, in sense orientation, to decrease lignin content, increase S/G lignin ratio and increase cellulose content compared to a non-transformed plant.
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72. A plurality of DNA constructs incorporated into the genome of angiosperms, the constructs comprising 4CL and CAD genes, substantially similar fragments thereof, and combinations thereof by sense or antisense orientation and CAld5H, AldOMT, and SAD genes, or substantially similar fragments thereof, and combinations thereof, in sense orientation, to decrease lignin content, increase S/G lignin ratio and increase cellulose content compared to a non-transformed plant.
Specification