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ENHANCED PRODUCTION OF TROPANE ALKALOIDS IN SCOPOLIA PARVIFLORA BY INTRODUCING THE PMT (PUTRESCINE N-METHYLTRANSFERASE) GENE

Lee, Ok-Sun (Division of Applied Life Science, Gyeongsang National University) , Kang, Young-Min (Division of Forest Science, Gyeongsang National University) , Jung, Hee-Young (Division of Forest Science, Gyeongsang National University) , Min, Ji-Yun (Division of Forest Science, Gyeongsang National University) , Kang, Seung-Mi (Division of Forest Science, Gyeongsang National University) , Chandrakant S. Karigar (Division of Biochemistry, Department of Chemistry, Central College Campus, Bangalore University) , D. Theertha Prasad (Department of Biotechnology, University of Agricultural Science, GKVK) , Bahk, Jung-Dong (Division of Applied Life Science, Gyeongsang National University) , Choi, Myung-Suk (Division of Forest Science, Gyeongsang National University, Environmental Biotechnology Research Center, Gyeongsang National University)

Plant molecular biology and biotechnology research center Annual report, 2005, 2005, 355-360

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In wild-type Scopolia parviflora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropane alkaloid levels were the highest in the root (0.9mg g^(-1) on a dry weight...
In wild-type Scopolia parviflora (Solanaceae) tissues, only the roots express the enzyme putrescine N-methyltransferase (PMT; EC 2.1.1.53), which is the first specific precursor of the tropane alkaloids. Moreover, the tropane alkaloid levels were the highest in the root (0.9mg g^(-1) on a dry weight basis), followed by the stem and then the leaves. We metabolically engineered S. parviflora by introducing the tobacco pmt into its genome by a binary vector system that employs disarmed Agrobacterium rhizogenes. The kanamycin-resistant hairy root lines were shown to bear the pmt gene and to overexpress its mRNA and protein product by at least two-fold, as determined polymerase chain reaction (PCR) and Northern and Western blottings, respectively. The transgenic lines also showed higher PMT activity and were morphologically aberrant in terms of slower growth and the production of lateral roots. The overexpression of pmt markedly elevated the scopolamine and hyoscyamine levels in the transgenic lines that showed the highest pmt mRNA and PMT protein levels. Thus, overexpression of the upstream regulator of the tropane alkaloid pathway enhanced the biosynthesis of the final product. These observations may be useful in establishing root culture systems that generate large yields of tropane alkaloids.