Nucleic acid sequences encoding transcription factors regulating alkaloid biosynthesis and their use in modifying plant metabolism
View Patent ↗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. A method for increasing a nicotinic alkaloid in a Nicotiana plant, comprising
introducing into a Nicotiana plant a vector comprising a nucleotide sequence selected from the group consisting of:
(i) the nucleotide sequence set forth in SEQ ID NO: 8 or SEQ ID NO: 9;
(ii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10;
(iii) a nucleotide sequence that is at least 90% identical to the nucleotide sequences of (i) or (ii), and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis; and
growing the plant under conditions which allow for the expression of a transcription factor that positively regulates nicotinic alkaloid biosynthesis from the nucleotide sequence;
wherein expression of the transcription factor results in the plant having an increased nicotinic alkaloid content as compared to a control plant grown under similar conditions.
2. The method of claim 1 , further comprising overexpressing within the Nicotiana plant at least one of NBB1, A622, quinolate phosphor-ribosyltransferase (Qpt), putrescine-N-methyltransferase (PMT), and N-methylputrescine oxidase (MPO).
3. The method of claim 1 , further comprising overexpressing within the Nicotiana plant at least one additional transcription factor that positively regulates nicotinic alkaloid biosynthesis.
4. The method of claim 1 , further comprising suppressing the expression of at least one transcription factor that negatively regulates nicotinic alkaloid biosynthesis within the Nicotiana plant.
5. The method of claim 1 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 8.
6. The method of claim 1 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 9.
7. The method of claim 1 , wherein the vector comprises the nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10.
8. The method of claim 1 , wherein the vector comprises a nucleotide sequence that is
(a) at least 90% identical to (i) the nucleotide sequence set forth in SEQ ID NO: 8; (ii) the nucleotide sequence set forth in SEQ ID NO: 9; or (iii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10, and
(b) which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
9. The method of claim 2 , wherein NBB1 is overexpressed.
10. The method of claim 2 , wherein A622 is overexpressed.
11. The method of claim 2 , wherein QPT is overexpressed.
12. The method of claim 2 , wherein PMT is overexpressed.
13. The method of claim 2 , wherein MPO is overexpressed.
14. The method of claim 3 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 8.
15. The method of claim 3 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 9.
16. The method of claim 3 , wherein the vector comprises the nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10.
17. The method of claim 3 , wherein the vector comprises a nucleotide sequence that is (a) at least 90% identical to (i) the nucleotide sequence set forth in SEQ ID NO: 8; (ii) the nucleotide sequence set forth in SEQ ID NO: 9; or (iii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10, and (b) which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
18. The method of claim 4 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 8.
19. The method of claim 4 , wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 9.
20. The method of claim 4 , wherein the vector comprises the nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10.
21. The method of claim 4 , wherein the vector comprises a nucleotide sequence that is (a) at least 90% identical to (i) the nucleotide sequence set forth in SEQ ID NO: 8; (ii) the nucleotide sequence set forth in SEQ ID NO: 9; or (iii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 10, and (b) which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.