Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/50456
標題: Phosphomannose-isomerase as a selectable marker to recover transgenic orchid plants (Oncidium Gower Ramsey)
作者: Thiruvengadam, M.
楊長賢
Hsu, W.H.
Yang, C.H.
關鍵字: Agrobacaterium tumefaciens
Mannose
Oncidium
Positive selection
Protocorm-like bodies
agrobacterium-mediated transformation
pmi/mannose selection system
zea-mays l.
genetic-transformation
biolistic transformation
maize
plants
mannose
cells
regeneration
tumefaciens
期刊/報告no:: Plant Cell Tissue and Organ Culture, Volume 104, Issue 2, Page(s) 239-246.
摘要: A transformation method using the phosphomannose-isomerase (pmi) gene as a selectable marker was developed for orchid Oncidium Gower Ramsey. The pmi-gene, which converts mannose-6-phosphate to fructose-6-phosphate allowing for selection of transgenic plants on mannose selective medium. Genetically transformed plants of Oncidium were regenerated after cocultivating protocorm-like bodies with Agrobacterium tumefaciens strain GV3101 containing the vectors pEPYON-42P and pEPYON-42H with 35S::PMI and 35S::HPTII genes respectively. We observed that 35S::PMI (pEPYON-42P) produced high rate (27 plants) of mannose resistant transgenic plants compared to 35S::HPTII (pEPYON-42H) in which only fourteen hygromycin resistant transgenic plants were obtained. Mannose resistant transgenic plants were confirmed by PCR and Southern blot. The pmi gene expression in 35S::PMI (pEPYON-42P) transgenic plants was confirmed by RT-PCR. Furthermore, the duration of regeneration time of transgenic plants was significantly shorter in mannose selected system (4 months) than in hygromycin selected system (8 months). The pmi/mannose selection system is shown to be highly efficient for producing transgenic O. Gower Ramsey without using antibiotics or herbicides. For the first time, the pmi/mannose-based "positive" selection system has been used to obtain genetically engineered O. Gower Ramsey.
URI: http://hdl.handle.net/11455/50456
ISSN: 0167-6857
文章連結: http://dx.doi.org/10.1007/s11240-010-9827-0
Appears in Collections:生物科技學研究所

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