Modulating alkaloid profiles in
A non-naturally occurring or transgenic Nicotiana tabacum plant cell encompassing a heterologous polynucleotide that encodes arginine decarboxylase (ADC) and a method for producing such a cell are provided. Transgenic Nicotiana tabacum plants, plant parts, and plant tissues including these cells and plant materials and biomass produced by these cells and plants cells forming such a plant are provided. Materials produced by this plant, including plant leaves, flowers, seeds, extracts, and other plant biomass obtained from this plant including materials that have altered alkaloid levels are disclosed. Also provided are smoking articles including various tobacco products containing these materials.
1 . A transgenic Nicotiana tabacum plant cell comprising a heterologous polynucleotide sequence that encodes a heterologous polypeptide sequence selected from:
a) a polypeptide sequence having at least 97% sequence identity to SEQ ID NO: 2, wherein the amino acid at position 31 is G, 40 is L, 63 is C, 296 is E, 355 is S, 417 is S, 429 is Y, 438 is S, 652 is M, 658 is D, 661 is M, 700 is D, 707 is I, and 730 is V; and
b) a polypeptide sequence having at least 97.5% sequence identity to SEQ ID NO: 4, wherein the amino acid at position 12 is A, 37 is I, 87 is M, 151 is F, 317 is V, 378 is E, 429 is V, 530 is V, and 680 is V;
wherein the heterologous polynucleotide sequence is operably linked to a promotor directing expression of the heterologous polypeptide sequence.
2 . The transgenic Nicotiana tabacum plant cell according to claim 1 , wherein the heterologous polypeptide sequence is a polypeptide sequence having at least 97% sequence identity to SEQ ID NO: 2, wherein the amino acid at position 31 is G, 40 is L, 63 is C, 296 is E, 355 is S, 417 is S, 429 is Y, 438 is S, 652 is M, 658 is D, 661 is M, 700 is D, 707 is I, and 730 is V.
3 . The transgenic Nicotiana tabacum plant cell according to claim 1 , wherein the heterologous polypeptide sequence is a polypeptide sequence having at least 97.5% sequence identity to SEQ ID NO: 4, wherein the amino acid at position 12 is A, 37 is I, 87 is M, 151 is F, 317 is V, 378 is E, 429 is V, 530 is V, and 680 is V.
4 . A transgenic Nicotiana tabacum plant comprising a plurality of plant cells according to claim 1 .
5 . The transgenic Nicotiana tabacum plant according to claim 4 , wherein levels of nicotine and nornicotine are decreased in the plant compared to an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
6 . The transgenic Nicotiana tabacum plant according to claim 5 , wherein levels of nicotine and nornicotine are decreased in the plant by at least 20% compared to an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
7 . The transgenic Nicotiana tabacum plant according to claim 6 , wherein levels of anabasine and anatabine are from 90% to 110% the levels of anabasine and anatabine in an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
8 . The transgenic Nicotiana tabacum plant cell according to claim 1 , wherein the heterologous polypeptide sequence comprises SEQ ID NO: 2 or SEQ ID NO: 4.
9 . A plant material comprising a transgenic Nicotiana tabacum plant that comprises the plant cell according to claim 1 ,
wherein the plant material is a cured or dried plant material, and
wherein the cured or dried plant material is flue-cured, fire-cured, smoke-cured, sun-cured, or air-cured.
10 . A tobacco product or smoking article comprising the transgenic Nicotiana tabacum plant material according to claim 9 .
11 . A transgenic Nicotiana tabacum plant cell comprising a heterologous polynucleotide sequence that encodes a heterologous polypeptide sequence having at least at least 97.5% sequence identity to SEQ ID NO: 6, wherein the amino acid at position 90 is M, 203 is F, 296 is E, 342 is V, 380 is Q, 441 is G, 619 is N, 628 is S, 663 is D, 664 is H, 680 is L, 698 is S, and 715 is S;
wherein the heterologous polynucleotide sequence is operably linked to a promotor directing expression of the heterologous polypeptide sequence.
12 . A transgenic Nicotiana tabacum plant comprising a plurality of plant cells according to claim 11 .
13 . The transgenic Nicotiana tabacum plant according to claim 12 , wherein levels of nicotine and nornicotine are increased in the plant compared to an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
14 . The transgenic Nicotiana tabacum plant according to claim 13 , wherein levels of nicotine and nornicotine are increased in the plant by at least 20% compared to an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
15 . The transgenic Nicotiana tabacum plant according to claim 14 , wherein levels of anabasine and anatabine are increased by at least 25% relative to the levels of anabasine and anatabine in an otherwise identical control Nicotiana tabacum plant not comprising cells comprising the heterologous polynucleotide.
16 . The transgenic Nicotiana tabacum plant cell according to claim 11 , wherein the heterologous polypeptide sequence comprises SEQ ID NO: 6.
17 . A plant material comprising a transgenic Nicotiana tabacum plant that comprises the plant cell according to claim 11 ,
wherein the plant material is a cured or dried plant material, and
wherein the cured or dried plant material is flue-cured, fire-cured, smoke-cured, sun-cured, or air-cured.
18 . A tobacco product or smoking article comprising the transgenic Nicotiana tabacum plant material according to claim 17 .