Agrobacterium-mediated transformation of Codonopsis lanceolata using the γ-TMT gene

Bimal Kumar Ghimire, Eun Soo Seong, Jung Dae Lim, Kweon Heo, Myong Jo Kim, Ill Min Chung, John A. Juvik, Chang Yeon Yu

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)265-274
Number of pages10
JournalPlant Cell, Tissue and Organ Culture
Volume95
Issue number3
DOIs
StatePublished - Dec 1 2008

Keywords

  • Agrobacterium tumefaciens
  • Codonopsis lanceolata
  • HPLC
  • Regeneration
  • Transformation
  • Transgenic
  • γ-Tocopherol

ASJC Scopus subject areas

  • Horticulture

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  • Cite this

    Ghimire, B. K., Seong, E. S., Lim, J. D., Heo, K., Kim, M. J., Chung, I. M., Juvik, J. A., & Yu, C. Y. (2008). Agrobacterium-mediated transformation of Codonopsis lanceolata using the γ-TMT gene. Plant Cell, Tissue and Organ Culture, 95(3), 265-274. https://doi.org/10.1007/s11240-008-9440-7