This disclosure provides a number of sequences involved in the transport of alkaloids from the root to the leaf in tobacco, methods of using such sequences, tobacco plants carrying modifications to such sequences, and tobacco products made from such plants.

A method of preparing a tobacco material for use in a smokeless tobacco product is provided, the method including receiving a tobacco material, wherein the tobacco material has not been subjected to a fermentation process, blending the tobacco material with the final moist snuff product ingredients to form a tobacco composition having a moisture content of at least about 50%, packaging the tobacco composition in a final product packaging such that after a fermentation period of 7 to 28 days within the final product packaging the product is ready for direct sale and consumption.

A method of preparing a tobacco material for use in a smokeless tobacco product is provided, the method including receiving a tobacco material, wherein the tobacco material has not been subjected to a fermentation process, blending the tobacco material with the final moist snuff product ingredients to form a tobacco composition having a moisture content of at least about 50%, packaging the tobacco composition in a final product packaging such that after a fermentation period of 7 to 28 days within the final product packaging the product is ready for direct sale and consumption.

The present invention features tobacco nicotine demethylase nucleic acid and amino acid sequences, tobacco plants and plant components containing such sequences, including tobacco plants and plant components having reduced expression or altered enzymatic activity of nicotine demethylase, methods of use of nicotine demethylase sequences to create plants having altered levels of nornicotine or N-nitrosonornicotine (NNN) or both relative to a control plant, as well as tobacco articles having reduced levels of nornicotine or NNN.

The present invention features tobacco nicotine demethylase nucleic acid and amino acid sequences, tobacco plants and plant components containing such sequences, including tobacco plants and plant components having reduced expression or altered enzymatic activity of nicotine demethylase, methods of use of nicotine demethylase sequences to create plants having altered levels of nornicotine or N-nitrosonornicotine (NNN) or both relative to a control plant, as well as tobacco articles having reduced levels of nornicotine or NNN.

Provided herein are bacterial strains that are capable of degrading nitrite and/or TSNAs. Also provided herein are methods of using such bacterial strains to degrade nitrite and/or TSNAs.

Provided herein are bacterial strains that are capable of degrading nitrite and/or TSNAs. Also provided herein are methods of using such bacterial strains to degrade nitrite and/or TSNAs.

There is described herein a mutant, non-naturally occurring or transgenic plant or part thereof having reduced expression or activity of asparagine synthetase, said asparagine synthetase comprising, consisting or consisting essentially of: (i) a polynucleotide sequence comprising, consisting or consisting essentially of a sequence having at least 90% sequence identity to SEQ ID NO:1 or at least 72% sequence identity to SEQ ID NO:3 or SEQ ID NO:5 or SEQ ID NO:7; or (ii) a polypeptide encoded by the polynucleotide set forth in (i); or (iii) a polypeptide having at least 78% sequence identity to SEQ ID NO:2 or SEQ ID NO:4 or SEQ ID NO:6 or SEQ ID NO:8; wherein the expression or activity of the asparagine synthetase set forth in (i), (ii) or (iii) is reduced as compared to a control plant.

There is described herein a mutant, non-naturally occurring or transgenic plant or part thereof having reduced expression or activity of asparagine synthetase, said asparagine synthetase comprising, consisting or consisting essentially of: (i) a polynucleotide sequence comprising, consisting or consisting essentially of a sequence having at least 90% sequence identity to SEQ ID NO:1 or at least 72% sequence identity to SEQ ID NO:3 or SEQ ID NO:5 or SEQ ID NO:7; or (ii) a polypeptide encoded by the polynucleotide set forth in (i); or (iii) a polypeptide having at least 78% sequence identity to SEQ ID NO:2 or SEQ ID NO:4 or SEQ ID NO:6 or SEQ ID NO:8; wherein the expression or activity of the asparagine synthetase set forth in (i), (ii) or (iii) is reduced as compared to a control plant.

Methods of forming cellulosic sugars from the stalk or roots of a plant of the Nicotiana species are provided herein, the methods including i) receiving a tobacco material including at least one of a stalk material and a root material of a harvested plant of the Nicotiana species; ii) delignifying the tobacco material to form a tobacco-derived pulp; and iii) hydrolyzing the tobacco-derived pulp to form a hydrolyzed tobacco product including residual solids and a liquid including at least one tobacco-derived cellulosic sugar. A cellulosic sugar derived from hydrolyzed tobacco material including at least one of a tobacco stalk material and a tobacco root material of a harvested plant of the Nicotiana species is also provided herein. Tobacco products incorporating the cellulosic sugars derived from hydrolyzed tobacco material or products derived from the cellulosic sugar are also provided.