Modification of plant lignin content
DNA constructs comprising a first DNA segment that corresponds to at least a portion of a gene in the monolignol biosynthetic pathway, a spacer DNA segment, and a second DNA segment that is complementary to the first DNA segment can be used to reduce or modulate the lignin content in plants. In some embodiments, DNA constructs comprise at least a portion of a gene for 4CL, C3H, CCR, C4H or CCoAOMT. Vascular-preferred and constitutive promoters can be used to drive expression of the constructs.
1. A DNA construct comprising a promoter operably linked to
a first DNA segment comprising from 200 bp to 600 bp of a Eucalyptus 4-coumarate co-enzyme A ligase (4CL) cDNA of SEQ ID NO: 65,
an intron spacer DNA segment comprising SEQ ID NO: 64, and
a second DNA segment that is fully complementary to the first DNA segment,
wherein the first and second DNA segments are arranged in a 5′ to 3′ direction with respect to each other.
2. The DNA construct of claim 1 wherein said promoter is a tissue-specific promoter.
3. The DNA construct of claim 2 wherein said promoter directs expression in a vascular-preferred manner such that expression is found in the xylem of plants.
4. The DNA construct of claim 1 wherein said promoter is a 4CL promoter from P. taeda.
5. The DNA construct of claim 1 wherein said first DNA segment has a fragment length of 200 bp.
6. The DNA construct of claim 1 wherein said first DNA segment comprises the nucleotide sequence of SEQ ID NO: 33.
7. The DNA construct of claim 1 wherein said DNA construct is pARB345 (ATCC Accession No. PTA-6223).
8. The DNA construct of claim 1 further comprising T-DNA border sequences.
9. A plant cell transformed with the DNA construct of claim 1 , wherein the plant cell expresses a double-stranded RNA encoded by the DNA construct.
10. A plant comprising the plant cell of claim 9 , wherein expression of the double-stranded RNA in the plant results in the down regulation of a 4CL (4-coumarate co-enzyme A ligase) gene expression involved in the monolignol biosynthetic pathway in the plant.
11. A method of reducing lignin content in a plant by inhibiting the expression of a 4CL (4-coumarate co-enzyme A ligase) gene involved in the monolignol biosynthetic pathway in the plant, comprising transforming a plant cell with the DNA construct of claim 1 , wherein the transformed plant cell expresses a double-stranded RNA encoded by the DNA construct, and culturing the transformed plant cell under conditions that promote regeneration of the transgenic plant, wherein expression of the double-stranded RNA in the plant results in the inhibition of the 4CL gene expression in the plant and thereby reducing lignin content in the transformed plant.