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 up-regulating a transcription factor that positively regulates nicotinic alkaloid biosynthesis, wherein the transcription factor is up-regulated by:
(a) introducing into the plant an expression construct comprising a nucleic acid molecule selected from the group consisting of:
(i) the nucleotide sequence set forth in SEQ ID NO: 11 or SEQ ID NO: 12;
(ii) a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 13; and
(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
(b) growing the plant under conditions whereby the expression construct increases levels of the transcription factor in the plant as compared to a control plant grown under similar conditions.
2. The method of claim 1 , further comprising up-regulating within the plant at least one of NBB1, A622, quinolate phospho-ribosyltransferase (QPT), putrescine-N-methyltransferase (PMT), and N-methylputrescine oxidase (MPO).
3. The method of claim 1 , further comprising up-regulating at least one additional transcription factor that positively regulates nicotinic alkaloid biosynthesis.
4. The method of claim 1 , further comprising down-regulating at least one transcription factor that negatively regulates nicotinic alkaloid biosynthesis.
5. The method of claim 1 , wherein the plant is a Nicotiana tabacum plant.
6. The method of claim 1 , wherein the expression construct comprises the nucleotide sequence set forth in SEQ ID NO: 11.
7. The method of claim 1 , wherein the expression construct comprises the nucleotide sequence set forth in SEQ ID NO: 12.
8. The method of claim 1 , wherein the expression construct comprises a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 13.
9. The method of claim 1 , wherein the expression construct comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 11, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
10. The method of claim 1 , wherein the expression construct comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 12, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.
11. The method of claim 1 , wherein the expression construct comprises a nucleotide sequence that is at least 90% identical to a nucleotide sequence that encodes a polypeptide having the amino acid sequence set forth in SEQ ID NO: 13, and which encodes a transcription factor that positively regulates nicotinic alkaloid biosynthesis.