An efficient Agrobacterium tumefaciens-mediated transformation and regeneration system for cotyledons of spinach (Spinacia oleracea L.) |
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Authors: | H-X Zhang J A D Zeevaart |
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Institution: | (1) MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing, MI 48824-1312, USA e-mail: zeevaart@pilot.msu.edu Fax: +1-517-353-9168, US |
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Abstract: | An efficient transformation and regeneration system was established for the production of transgenic spinach (Spinacia oleracea L.) plants. Cotyledon explants were infected with Agrobacterium tumefaciens strain LBA4404 carrying the selectable marker gene, neomycin phosphotransferase II (nptII), and the reporter gene smgfp, encoding soluble-modified green-fluorescent protein, driven by the cauliflower mosaic virus 35S promoter. The infected explants
were cultured on Murashige and Skoog medium, containing 1 mg/l benzyladenine and 0.4 mg/l naphthaleneacetic acid. Shoots were
regenerated on selection medium containing 50 mg/l kanamycin. Regenerated kanamycin-resistant shoots were rooted on medium
containing 1 mg/l indolebutyric acid and subsequently grown in soil in the greenhouse. Southern blot analysis indicated that
the smgfp gene had been integrated into the spinach genome. Northern and Western blots showed that the smgfp gene was expressed in progeny plants.
Received: 31 March 1998 / Revision received: 27 September 1998 / Accepted: 10 Ocotber 1998 |
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Keywords: | Spinach (Spinacia oleracea L ) Regeneration Green-fluorescent protein Transformation Cotyledon |
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