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 at least 21 consecutive nucleotides of a nucleotide sequence, wherein the nucleotide sequence is selected from the group consisting of:
(a) the nucleotide sequence set forth in SEQ ID NO: 4; and
(b) a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 4; and
wherein the 21 consecutive nucleotides are in sense orientation, antisense orientation, or both.
2. The method of claim 1 , further comprising overexpressing within the Nicotiana 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 overexpressing at least one transcription factor that positively regulates nicotinic alkaloid biosynthesis.
4. The method of claim 1 , further comprising suppressing the expression of at least one additional transcription factor that negatively regulates nicotinic alkaloid biosynthesis.
5. The method of claim 1 , wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 4.
6. The method of claim 1 , wherein the nucleotide sequence comprises a nucleotide sequence that is at least 90% identical to the nucleotide sequence set forth in SEQ ID NO: 4.
7. The method of claim 1 , wherein the nucleotide sequence comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence set forth in SEQ ID NO: 4.
8. The method of claim 2 , wherein NBB 1 is overexpressed.
9. The method of claim 2 , wherein A622 is overexpressed.
10. The method of claim 2 , wherein QPT is overexpressed.
11. The method of claim 2 , wherein PMT is overexpressed.
12. The method of claim 2 , wherein MPO is overexpressed.
13. A transgenic Nicotiana plant produced by the method of claim 1 and having an increased alkaloid content as compared to a control plant grown under similar conditions.
14. The transgenic Nicotiana plant of claim 13 , wherein the plant is a Nicotiana tabacum plant.
15. A product having an increased level of a nicotinic alkaloid comprising the transgenic Nicotiana plant of claim 13 , or portions thereof as compared to a product comprising a control plant or portions thereof.
16. Seeds from the transgenic Nicotiana plant of claim 13 , wherein the seeds comprise a vector comprising at least 21 consecutive nucleotides of a nucleotide sequence, wherein the nucleotide sequence is selected from the group consisting of:
(a) the nucleotide sequence set forth in SEQ ID NO: 4; and
(b) a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 4; and
wherein the 21 consecutive nucleotides are in sense orientation, antisense orientation, or both.
17. The product of claim 15 , wherein the product is selected from the group consisting of a cigarette, cigarette tobacco, cigar tobacco, a cigar, pipe tobacco, chewing tobacco, snuff, snus, lozenges, and reconstituted tobacco.
18. The method of claim 1 , wherein the nucleotide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 4.