<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Codonopsis lanceolata</Emphasis> using the <Emphasis Type="Italic">γ-TMT</Emphasis> gene |
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Authors: | Bimal Kumar Ghimire Eun Soo Seong Jung Dae Lim Kweon Heo Myong Jo Kim Ill-Min Chung John A Juvik Chang Yeon Yu |
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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; |
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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. |
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Keywords: | Agrobacterium tumefaciens Codonopsis lanceolata Transformation Transgenic Regeneration γ -Tocopherol HPLC |
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