Nucleic acid sequences to proteins involved in isoprenoid synthesis
Methods are provided for producing plants and seeds having altered isoprenoid content and compositions. The methods find particular use in increasing the isoprenoid levels in plants, and in providing desirable isoprenoid compositions in a host plant cell.
1. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, wherein said 1-deoxy-D-xylulose 5-phosphate reductoisomerase is found in Arabidopsis.
2. An isolated polynucleotide selected from the group consisting of:
(a) an isolated polynucleotide comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2;
(b) an isolated polynucleotide comprising SEQ ID NO: 1;
(c) an isolated polynucleotide comprising a nucleotide sequence which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase and which has at least 95% identity to that of SEQ ID NO: 1 over the entire length of SEQ ID NO: 1;
(d) an isolated polynucleotide that hybridizes under stringent conditions to SEQ ID NO: 1 over the entire length of SEQ ID NO: 1; said stringent conditions comprising overnight incubation at 42° C. in a solution comprising 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate and 20 micrograms/milliliter denatured, sheared salmon sperm DNA, followed by washing the hybridization support in 0.1×SSC at approximately 65° C.; and which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase; and
(e) an isolated polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), or (d).
3. A DNA construct comprising, as operably associated components in the 5′ to 3′ direction of transcription:
a promoter functional in a plant cell;
a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2;
(b) a polynucleotide comprising SEQ ID NO: 1;
(c) a polynucleotide comprising a nucleotide sequence which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase and which has at least 95% identity to that of SEQ ID NO: 1 over the entire length of SEQ ID NO: 1;
(d) an isolated polynucleotide that hybridizes under stringent conditions to SEQ ID NO: 1 over the entire length of SEQ ID NO: 1; said stringent conditions comprising overnight incubation at 42° C. in a solution comprising 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate and 20 micrograms/milliliter denatured, sheared salmon sperm DNA, followed by washing the hybridization support in 0.1×SSC at approximately 65° C.; and which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase; and
(e) a polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), or (d); and,
a transcriptional termination sequence.
4. A host cell comprising the DNA construct of claim 3 .
5. The host cell according to claim 4 , wherein the host cell is a plant cell.
6. A plant comprising a cell comprising a DNA construct comprising, as operably associated components in the 5′ to 3′ direction of transcription:
a promoter functional in a plant cell;
a polynucleotide selected from the group consisting of:
(a) a polynucleotide comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2;
(b) a polynucleotide comprising SEQ ID NO: 1;
(c) a polynucleotide comprising a nucleotide sequence which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase and which has at least 95% identity to that of SEQ ID NO: 1 over the entire length of SEQ ID NO: 1;
(d) an isolated polynucleotide that hybridizes under stringent conditions to SEQ ID NO: 1 over the entire length of SEQ ID NO: 1; said stringent conditions comprising overnight incubation at 42° C. in a solution comprising 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate and 20 micrograms/milliliter denatured, sheared salmon sperm DNA, followed by washing the hybridization support in 0.1×SSC at approximately 65° C.; and which encodes a 1-deoxy-D-xylulose 5-phosphate reductoisomerase; and,
(e) a polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), (d); and,
a transcriptional termination sequence.
7. An isolated polynucleotide according to claim 2 , wherein said polynucleotide comprises a nucleotide sequence encoding the polypeptide of SEQ ID NO: 2.
8. An isolated DNA comprising a nucleic acid which encodes an Arabidopsis 1-deoxy-D-xylulose-5-phsophate reductiosiomerase that has the amino acid sequence of SEQ ID NO:2.