Compositions and methods for xylem-specific expression in plant cells
View Patent ↗The invention provides promoter sequences that regulate specific expression of operably linked sequences in developing xylem cells and/or in developing xylem tissue. The developing xylem-specific sequences are exemplified by the DX5, DX8, DX11, and DX15 promoters, portions thereof, and homologs thereof. The invention further provides expression vectors, cells, tissues and plants that contain the invention's sequences. The compositions of the invention and methods of using them are useful in, for example, improving the quantity (biomass) and/or the quality (wood density, lignin content, sugar content etc.) of expressed biomass feedstock products that may be used for bioenergy, biorefinary, and generating wood products such as pulp, paper, and solid wood.
1. A recombinant nucleic acid comprising a first nucleic acid segment with at least 98% sequence identity to SEQ ID NO: 08 that is operably-linked to a heterologous second nucleic acid.
2. The nucleic acid of claim 1 , wherein said first nucleic acid segment drives developing xylem expression of the operably-linked second nucleic acid in a plant cell.
3. The nucleic acid of claim 2 , wherein said second nucleic acid encodes
(a) a polypeptide involved in biosynthesis of a polysaccharide selected from cellulose and lignin,
(b) an antisense RNA of a sequence involved in biosynthesis of a polysaccharide selected from cellulose and lignin, or
(c) an interference RNA (iRNA) of a sequence involved in biosynthesis of a polysaccharide selected from cellulose and lignin.
4. An expression vector comprising the recombinant nucleic acid of claim 1 .
5. A cell comprising the recombinant nucleic acid of claim 1 .
6. The cell of claim 5 , wherein said cell is selected from a bacterial cell, a mammalian cell, an insect cell, a plant cell, and a fungal cell.
7. The cell of claim 6 , wherein said plant cell is selected from a developing xylem cell and a seed cell.
8. A transgenic plant comprising the recombinant nucleic acid of claim 1 .
9. The transgenic plant of claim 8 , wherein said heterologous second nucleic acid encodes a sequence selected from the group of
(a) a polypeptide involved in biosynthesis of a polysaccharide selected from cellulose and lignin,
(b) an antisense RNA of a sequence involved in biosynthesis of a polysaccharide selected from cellulose and lignin, and
(c) an interference RNA (iRNA) of a sequence involved in biosynthesis of a polysaccharide selected from cellulose and lignin.
10. A seed comprising the recombinant nucleic acid of claim 1 .
11. A composition comprising xylem, wherein said xylem comprises the recombinant nucleic acid of claim 1 .
12. A method for expressing a polypeptide or RNA from a nucleic acid in a plant cell comprising
a) providing
i) an expression vector comprising the recombinant nucleic acid of claim 1 , wherein the second nucleic acid encodes a polypeptide or an RNA, and
ii) a plant cell,
b) introducing the expression vector into said plant cell to produce a transgenic plant cell, and culturing the transgenic plant cell under conditions for expression of said polypeptide or RNA from said second nucleic acid in said transgenic plant cell.
13. The method of claim 12 , further comprising c) detecting expression of said polypeptide or RNA from said second nucleic acid in said transgenic cell.
14. The method of claim 12 , wherein said second nucleic acid encodes
(a) a polypeptide involved in biosynthesis of a polysaccharide selected from cellulose and lignin,
(b) an antisense RNA of a sequence involved in biosynthesis of a polysaccharide selected froth cellulose and lignin, or
(c) an interference RNA (iRNA) of a sequence involved in biosynthesis of a polysaccharide selected from cellulose and lignin.
15. The method of claim 12 , further comprising c) regenerating a transgenic plant from said transgenic plant cell.
16. The method of claim 15 , further comprising d) detecting expression of said polypeptide or RNA from said second nucleic acid in a developing xylem cell of said transgenic plant.
17. The method of claim 12 , wherein the heterologous second nucleic acid encodes an enzyme.