An efficient Agrobacterium tumefaciens mediated transformation and regeneration system for cotyledons of spinach (Spinacia oleracea L)

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 carryi ng the selectable marker gene, neomycin phospotransferase II (nptII), and t he 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 benzyladeni ne and 0.4 mg/l naphthaleneacetic acid. Shoots were regenerated on selectio n medium containing 50 mg/l kanamycin. Regenerated kanamycin-resistant shoo ts were rooted on medium containing 1 mg/l indolebutyric acid and subsequen tly grown in soil in the greenhouse. Southern blot analysis indicated chat the smgfp gene bad been integrated into the spinach genome. Northern and We stern blots showed that the smgfp gene was expressed in progeny plants.