Efficient production of transgenic plantls of Vanda through sonication-assisted Agrobacterium-mediated transformation of protocorm-like bodies
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- Shrestha Bipna Rani
- Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University
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- Chin Dong Poh
- Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University
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- Tokuhara Ken
- Research and Development Center, Orchid Resort Dogashima
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- Mii Masahiro
- Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University
Bibliographic Information
- Other Title
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- Transgenic note: Efficient production of transgenic plants of Vanda through sonication-assisted Agrobacterium-mediated transformation of protocorm-like bodies
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Abstract
In Vanda orchids, it is important to produce cultivars with economically important traits such as disease and pest resistances and novel flower colors, which are difficult to achieve by conventional cross breeding methods. To realize these breeding objectives, it is now expected to apply genetic transformation technology to introduce useful foreign genes into Vanda orchids. However, there has been almost no information on the genetic transformation of Vanda. Transgenic plants were successfully regenerated after co-cultivating protocorm-like bodies (PLBs) with Agrobacterium tumefaciens strain EHA101 (pIG121Hm) that harbored genes for β-glucuronidase (gus), hygromycin phosphotransferase (hpt) and neomycin phosphotransferase II (nptII). PLBs of ‘Tokyo Blue’ maintained in liquid New Dogashima medium (NDM) under dark condition, were subjected to transformation experiments. The PLBs inoculated with Agrobacterium produced secondary PLBs 4 weeks after transfer onto 3 g l−1 gellan gum-solidified NDM containing 30 g l−1 maltose, 10 mg l−1 meropenem and 10 mg l−1 hygromycin. Transformation efficiency was increased by prolonged period of infection (240 min) and wounding treatment of PLBs by sonication (5 min) during inoculation. Transformation of the hygromycin-resistant plantlets regenerated from different PLBs was confirmed by histochemical GUS assay, PCR analysis and Southern hybridization. With this transformation system, approximately 17 independent transgenic plants were obtained from 1 g PLBs.
Journal
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- Plant Biotechnology
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Plant Biotechnology 24 (4), 429-434, 2007
Japanese Society for Plant Biotechnology
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Details 詳細情報について
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- CRID
- 1390282679304480128
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- NII Article ID
- 10021911987
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- NII Book ID
- AA11250821
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- COI
- 1:CAS:528:DC%2BD2sXhtF2rtrvN
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- ISSN
- 13476114
- 13424580
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- NDL BIB ID
- 8905177
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed