Nucleic acid sequences encoding transcription factors regulating alkaloid biosynthesis and their use in modifying plant metabolism
Plant metabolism and alkaloid levels can be regulated by transcription factors that regulate the nicotinic alkaloid biosynthetic pathway. In one embodiment, the disclosure provides a transcription factor that negatively regulates alkaloid biosynthesis, such as nicotine biosynthesis.
1. An isolated cDNA molecule comprising a nucleotide sequence selected from the group consisting of:
(a) the nucleotide sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2;
(b) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3; and
(c) a nucleotide sequence that is at least 90% identical to the nucleotide sequences of (a) or (b), and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
2. An expression vector comprising the cDNA molecule of claim 1 , operably linked to one or more control sequences suitable for directing expression in a Nicotiana host cell.
3. A genetically engineered nicotinic alkaloid-producing Nicotiana plant comprising a cell comprising a chimeric nucleic acid construct comprising the isolated cDNA molecule of claim 1 , wherein the isolated cDNA molecule is linked to a heterologous nucleic acid to form the chimeric nucleic acid construct.
4. The engineered Nicotiana plant of claim 3 , wherein the plant is a Nicotiana tabacum plant.
5. Seeds from the engineered Nicotiana plant of claim 3 , wherein the seeds comprise the chimeric nucleic acid construct.
6. A product comprising the engineered Nicotiana plant of claim 3 .
7. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is set forth in SEQ ID NO: 1.
8. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is set forth in SEQ ID NO: 2.
9. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3.
10. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 90% identical to the nucleotide sequence of SEQ ID NO: 1, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
11. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
12. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 90% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
13. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 95% identical to the nucleotide sequence of SEQ ID NO: 1, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
14. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 95% identical to the nucleotide sequence of SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
15. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 95% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
16. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 96% identical to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
17. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 96% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
18. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 97% identical to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
19. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 97% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
20. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 98% identical to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
21. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 98% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
22. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 99% identical to the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
23. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence is at least 99% identical to the nucleotide sequence encoding a polypeptide having the amino acid sequence set forth in SEQ ID NO: 3, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.