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

作者：Ghimire Bimal Kumar; Seong Eun Soo; Lim Jung Dae; Heo Kweon; Kim Myong Jo; Chung Ill Min; Juvik John A; Yu Chang Yeon^*

来源：Plant Cell, Tissue and Organ Culture, 2008, 95(3): 265-274.

DOI：10.1007/s11240-008-9440-7

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

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 alpha-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 gamma-tocopherol methyltransferase (gamma-TMT) by this protocol. Moreover, the alpha-tocopherol content of the plants was enhanced by the overexpression of this gene.

• 出版日期2008-12