Transcription factors that regulate nicotine biosynthesis in tobacco
The present invention provides nucleic acids encoding transcription factors and methods of using these nucleic acids to modulate nicotine production in plants and to produce plants having modulated nicotine production.
1. An isolated nucleic acid molecule comprising a full length cDNA sequence selected from the group consisting of:
(a) the nucleotide sequence of SEQ ID NO:1; and
(b) a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO:1, wherein the nucleotide sequences of (a) and (b) are transcription factors that modulate alkaloid content, which when overexpressed produce an alkaloid content of at least about 4.5% of dry weight in a field grown transgenic Nicotiana tabacum plant post topping.
2. A nucleic acid construct comprising in the 5 to 3′ direction, a heterologous promoter operable in a plant cell and positioned downstream from said promoter and operatively associated therewith a nucleic acid molecule selected from the group consisting of:
(a) the nucleotide sequence of SEQ ID NO: 1; and
(b) a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO:1, wherein the nucleotide sequences of (a) and (b) are transcription factors that modulate alkaloid content, which when overexpressed produce an alkaloid content in a field grown transgenic Nicotiana tabacum plant post topping of at least about 4.5% of dry weight.
3. The nucleic acid construct of claim 2 , wherein the promoter is a constitutive promoter, a tissue preferred promoter or a tissue specific promoter.
4. A vector comprising the nucleic acid molecule of claim 1 .
5. A transgenic Nicotiana tabacum plant cell or bacterial cell comprising the vector of claim 4 .
6. A transgenic Nicotiana tabacum plant comprising the plant cell of claim 5 .
7. A seed of the transgenic plant of claim 6 or a progeny plant produced from the seed, wherein the seed and progeny plant comprise in their genomes said vector.
8. A crop comprising a plurality of transgenic Nicotiana tabacum plants according to claim 6 , or a transgenic progeny thereof, wherein the plurality of transgenic plants are planted together in an agricultural field, and the transgenic N. tabacum plants and transgenic progeny thereof comprise in their genomes said vector.
9. A tobacco product produced from the transgenic Nicotiana tabacum plant of claim 6 , wherein the tobacco product comprises said vector.
10. The tobacco product of claim 9 , wherein the tobacco product is selected from the group consisting of a cigarette, cigarette tobacco, cigar tobacco, a cigar, pipe tobacco, chewing tobacco, leaf tobacco, shredded tobacco, cut tobacco, and any combination thereof.
11. A method of producing a transgenic Nicotiana tabacum plant having increased alkaloid content, comprising
introducing into a Nicotiana tabacum plant cell a nucleic acid construct comprising a nucleotide sequence that encodes a transcription factor having the amino acid sequence of SEQ ID NO:2 to produce a transgenic Nicotiana tabacum plant cell comprising said nucleic acid construct; and
regenerating said transgenic Nicotiana tabacum plant cell to produce a transgenic Nicotiana tabacum plant comprising said nucleic acid construct, wherein leaf lamina of said transgenic Nicotiana tabacum plant when field grown have an alkaloid content of at least about 4.5% of dry weight post topping.
12. A transgenic Nicotiana tabacum plant produced by the method of claim 11 , wherein the transgenic N. tabacum plant comprises in its genome the nucleotide sequence that encodes the transcription factor having the amino acid sequence of SEQ ID NO:2.
13. A crop comprising a plurality of transgenic Nicotiana tabacum plants of claim 12 , or a transgenic progeny plant thereof, wherein the plurality of transgenic plants are planted together in an agricultural field and the transgenic N. tabacum plant and transgenic progeny plant thereof comprise in their genomes the nucleotide sequence that encodes the transcription factor having the amino acid sequence of SEQ ID NO:2.
14. A tobacco product produced from the transgenic Nicotiana tabacum plant of claim 12 , wherein the tobacco product comprises the nucleotide sequence that encodes the transcription factor having the amino acid sequence of SEQ ID NO:2.
15. The tobacco product of claim 14 , wherein the tobacco product is selected from the group consisting of a cigarette, cigarette tobacco, cigar tobacco, a cigar, pipe tobacco, Chewing tobacco, leaf tobacco, shredded tobacco, cut tobacco, and any combination thereof.