Patent ID: 8624083

Claim:
A method for increasing nicotine levels in a population of Nicotiana plants, comprising: (a) providing a population of mutated Nicotiana plants; (b) detecting and selecting a target mutated Nicotiana plant within said population, wherein said detecting comprises: (i) using primers developed from SEQ ID NO: 14 to amplify regions of a transcription factor gene from mutated Nicotiana plants in the population of mutated Nicotiana plants; (ii) identifying one or more mismatch between the amplified regions and corresponding regions in a wild-type gene, wherein the wild-type gene is from a non-mutated Nicotiana plant, and encodes a transcription factor that negatively regulates nicotine biosynthesis having an amino acid sequence that is at least 90% identical to SEQ ID NO: 16; and (iii) identifying the mutated Nicotiana plant that contains the one or more mismatch, wherein said identified target mutated Nicotiana plant has decreased expression of a transcription factor that negatively regulates nicotine biosynthesis and increased nicotine levels, compared to a control Nicotiana plant comprising the wild type gene; and (c) selectively breeding the target mutated Nicotiana plant and producing a population of plants having decreased expression of the transcription factor that negatively regulates nicotine biosynthesis and increased nicotine levels, compared to a population of control plants.