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<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Codonopsis lanceolata</Emphasis> using the <Emphasis Type="Italic">γ-TMT</Emphasis> gene
Authors:Bimal Kumar Ghimire  Eun Soo Seong  Jung Dae Lim  Kweon Heo  Myong Jo Kim  Ill-Min Chung  John A Juvik  Chang Yeon Yu
Institution:(1) Bioherb Research Institute, Kangwon National University, Chuncheon, 200-701, South Korea;(2) Department of Herbal Medicine Resource, Kangwon National University, Samcheok, 245-711, South Korea;(3) Division of Bio-resources Technology, College of Agriculture and Life Science, Kangwon National University, Chuncheon, 200-701, South Korea;(4) Department of Applied Life Science, Konkuk University, Seoul, 143-701, South Korea;(5) Department of Natural Resources and Environmental Sciences, University of Illinois, Urbana, IL 61801, USA;
Abstract:Efficient transformation of leaf disc-derived callus of Codonopsis lanceolata was obtained using Agrobacterium tumefaciens strain LBA4404 harboring a binary vector, pYBI121, that carries the neomycin phosphotransferase (npt II) gene as a selectable marker. The green shoots recovered from agroinfected explants on selection medium (containing 0.1 mg/l α-naphthaleneacetic acid (NAA), 1 mg/l 6-benzylaminopurine (BAP), 100 mg/l kanamycin, and 250 mg/l cefotaxime) were rooted on Murashige and Skoog (MS) medium supplemented with 2 mg/l IBA and 10 mg/l kanamycin. To optimize the transformation conditions, several factors were assessed, including the co-cultivation period, the duration of pre- and post-culture in darkness and light, the kanamycin concentration, and the Agrobacterium densities. We produced transgenic Codonopsis lanceolata overexpressing γ-tocopherol methyltransferase (γ-TMT) by this protocol. Moreover, the α-tocopherol content of the plants was enhanced by the overexpression of this gene. Bimal Kumar Ghimire and Eun Soo Seong contributed equally to this work.
Keywords:Agrobacterium tumefaciens                      Codonopsis lanceolata            Transformation  Transgenic  Regeneration  γ  -Tocopherol  HPLC
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