Various embodiments are directed to consumable products incorporating a matrix of cultured tobacco cells. Any tobacco variety may be utilized for establishing in vitro tobacco cultures, including native tobacco varieties and genetically modified tobacco varieties derived from any tobacco variety. Various embodiments are directed to methods for producing tobacco products incorporating cellular material and/or extracts derived from tobacco cells cultured in vitro.

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.

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.

Methods of modifying the tobacco-specific nitrosamine content of a tobacco material are described herein. One exemplary method comprises contacting a tobacco material with a composition comprising salt, sugar, enzyme, lactic acid bacteria, yeast, or a combination thereof to reduce the total bacterial content; curing the tobacco material; and fermenting the tobacco material in the presence of one or more microorganisms. The method can provide a fermented tobacco material having a tobacco-specific nitrosamine content that is reduced relative to a fermented tobacco material that has not been subjected to the disclosed method steps. In certain embodiments, the tobacco-specific nitrosamine content of the fermented tobacco material is no more than that of the cured tobacco material. Tobacco-containing products including such treated tobacco materials are also provided.

Methods of modifying the tobacco-specific nitrosamine content of a tobacco material are described herein. One exemplary method comprises contacting a tobacco material with a composition comprising salt, sugar, enzyme, lactic acid bacteria, yeast, or a combination thereof to reduce the total bacterial content; curing the tobacco material; and fermenting the tobacco material in the presence of one or more microorganisms. The method can provide a fermented tobacco material having a tobacco-specific nitrosamine content that is reduced relative to a fermented tobacco material that has not been subjected to the disclosed method steps. In certain embodiments, the tobacco-specific nitrosamine content of the fermented tobacco material is no more than that of the cured tobacco material. Tobacco-containing products including such treated tobacco materials are also provided.

A method of modifying the content of certain bacteria in uncured tobacco material is provided, the method including contacting an uncured tobacco material with a treatment solution, wherein the treatment solution is selected from the group consisting of: (i) a solution comprising salt, sugar, or a combination thereof; (ii) a solution comprising one or more enzymes; and (iii) a solution comprising one or more probiotics, wherein said contacting provides a treated tobacco material having a reduced total bacterial content following harvest. Also provided is a method of reducing TSNA content, the method including growing a tobacco plant in soil treated with a chloride source; harvesting the tobacco plant; and curing the harvested tobacco plant to give a cured tobacco material, providing a treated, cured tobacco material having a TSNA content that is reduced as compared to a cured tobacco material that has been grown in soil not treated with a chloride source. Smoking articles and smokeless tobacco products including such treated tobacco materials are also provided.

A method of modifying the content of certain bacteria in uncured tobacco material is provided, the method including contacting an uncured tobacco material with a treatment solution, wherein the treatment solution is selected from the group consisting of: (i) a solution comprising salt, sugar, or a combination thereof; (ii) a solution comprising one or more enzymes; and (iii) a solution comprising one or more probiotics, wherein said contacting provides a treated tobacco material having a reduced total bacterial content following harvest. Also provided is a method of reducing TSNA content, the method including growing a tobacco plant in soil treated with a chloride source; harvesting the tobacco plant; and curing the harvested tobacco plant to give a cured tobacco material, providing a treated, cured tobacco material having a TSNA content that is reduced as compared to a cured tobacco material that has been grown in soil not treated with a chloride source. Smoking articles and smokeless tobacco products including such treated tobacco materials are also provided.

The present invention relates to a tobacco plant, etc. This tobacco plant has either one or both of the following: (i) an endogenous gene containing, as an encoding region, a nucleic acid containing a base sequence encoding SEQ ID NO: 2 or an amino acid sequence having at least 95% identity with SEQ ID NO: 2, in which at least one codon corresponding to the amino acid sequence has mutated to a stop codon; and (ii) an endogenous gene containing, as an encoding region, a nucleic acid containing a base sequence encoding SEQ ID NO: 4 or an amino acid sequence having at least 95% identity with SEQ ID NO: 4, in which at least one codon corresponding to the amino acid sequence has mutated to a stop codon.

The present invention relates to a tobacco plant, etc. This tobacco plant has either one or both of the following: (i) an endogenous gene containing, as an encoding region, a nucleic acid containing a base sequence encoding SEQ ID NO: 2 or an amino acid sequence having at least 95% identity with SEQ ID NO: 2, in which at least one codon corresponding to the amino acid sequence has mutated to a stop codon; and (ii) an endogenous gene containing, as an encoding region, a nucleic acid containing a base sequence encoding SEQ ID NO: 4 or an amino acid sequence having at least 95% identity with SEQ ID NO: 4, in which at least one codon corresponding to the amino acid sequence has mutated to a stop codon.

Two genes, A622 and NBB1, can be influenced to achieve a decrease of nicotinic alkaloid levels in plants. In particular, suppression of one or both of A622 and NBB1 may be used to decrease nicotine in tobacco plants.