Nicotiana nucleic acid molecules and uses thereof

A method of breeding a tobacco plant, said method comprising the steps of:

• a) crossing a plant of a first tobacco line with a plant of a second tobacco line to produce F1 progeny tobacco plants, said first tobacco plant having an endogenous nucleic acid comprising a mutation in the nucleotide sequence set forth in SEQ ID NO:
• 4;
• b) extracting a DNA sample from each of a plurality of said F1 progeny tobacco plants;
• c) contacting each said DNA sample with a marker nucleic acid that hybridizes to said endogenous nucleic acid or to a fragment thereof;
• d) screening said DNA samples to identify F1 plants having said mutation;
• and
• e) screening F1 plants having said mutation for plants exhibiting reduced expression of nicotine demethylase relative to plants lacking said mutation.

A method of producing tobacco seed, comprising

• a) crossing a plant of a first tobacco line with a plant of a second tobacco line, said first tobacco plant selected from the group consisting of Nicotiana africana, Nicotiana amplexicaulis, Nicotiana arentsii, Nicotiana benthamiana, Nicotiana bigelovii, Nicotiana corymbosa, Nicotiana debneyi, Nicotiana excelsior, Nicotiana exigua, Nicotiana glutinosa, Nicotiana goodspeedii, Nicotiana gossei, Nicotiana hesperis, Nicotiana ingulba, Nicotiana knightiana, Nicotiana maritima, Nicotiana megalosiphon, Nicotiana miersii, Nicotiana nesophila, Nicotiana noctiflora, Nicotiana nudicaulis, Nicotiana otophora, Nicotiana palmeri, Nicotiana paniculata, Nicotiana petunioides, Nicotiana plumbaginifolia, Nicotiana repanda, Nicotiana rosulata, Nicotiana rotundifolia, Nicotiana rustica, Nicotiana setchelli, Nicotiana stocktonii, Nicotiana eastii, and Nicotiana suaveolens, and said second tobacco plant having an endogenous nucleic acid comprising a mutation in the nucleotide sequence set forth in SEQ ID NO:
• 4;
• and
• b) harvesting seed produced from said cross.