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Overexpression of Hyoscyamine 6β-Hydroxylase (h6h) Gene and Enhanced Production of Tropane Alkaloids in Scopolia parviflora Hairy Root Lines

Kang, Young-Min (Division of Forest Science, Gyeongsang National University) , Lee, Ok-Sun (Division of Applied Life Science, Gyeongsang National University) , Jung, Hee-Young (Division of Forest Science, Gyeongsang National University) , Kang, Seung-Mi (Division of Forest Science, Gyeongsang National University) , Lee, Byung-Hyun (Division of Applied Life Science, Gyeongsang National University) , Chandrakant Karigar (Division of Biochemistry, Department of Chemistry, Central College Campus, Bangalore University) , 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, 361-368

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The hyoscyamine 6β-hydroxylase (h6h) gene was introduced into the genome of Scopolia parviflora through the Agrobacterium rhizogenes binary vector system. The enzyme was expressed ally and tissue specific selectively in roots, resulting in five transgenic hairy root lines. The presence of the h6h ge...
The hyoscyamine 6β-hydroxylase (h6h) gene was introduced into the genome of Scopolia parviflora through the Agrobacterium rhizogenes binary vector system. The enzyme was expressed ally and tissue specific selectively in roots, resulting in five transgenic hairy root lines. The presence of the h6h gene in kanamycin-resistant hairy roots and its overexpression were confirmed by polymerase chain reaction (PCR), Northern blotting, and Western blotting, respectively. In the transgenic hairy root lines which constitutively expressed the H6H enzyme, hyoscyamine and scopolamine accumulated in high concentration. Among the transgenic hairy root lines that expressed the H6H enzyme, only two were more productive. The levels of tropane alkaloids in transgenic hairy root varied greatly: The best transgenic line (#5) contained 8.12 mg of scopolamine per g dry weight, which produced the compoung three times more than wild-type root. These results suggest a possibility of improving the yield of tropane alkaloids in hairy root lines by genetic and metabolic engineering.