The invention relates to a PMT2 gene promoter for nicotine biosynthesis and an application thereof. The PMT2 gene promoter for nicotine biosynthesis is extracted from an N. sylvestris genome sequence, and has a nucleotide sequence as shown in SEQ ID NO.1. The promoter region of the nicotine PMT2 gene can generate transcription with a basic level, and the regulation region can response to various ambient conditions and thus regulate the expression level of the nicotine synthesis genes correspondingly (up-regulation of down-regulation as required), which has important practical significance on the production of nicotine extraction raw materials with high quality.
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