Biolistic transformation of rose (Rosa hybrida L.)

Abstract

A reproducible method has been developed for the Biolistic transformation and regeneration of transgenic plants from embryogenic callus of rose (Rosa hybrida L.) cv. Glad Tidings. DNA delivery was optimized using the β-glucuronidase (gus) gone. The distance between the stopping screen and target explants and supplementation of pre-and post-bombardment culture media with 0.25 M myo-inositol influenced the transformation efficiency. Prior to culture on selection medium containing 250 mg l-1 kanamycin sulphate, embryogenic calli were bombarded, using optimized gene delivery parameters, with a plasmid carrying the neomycin phosphotransferase (npt II) gene. Somatic embryo-derived kanamycin-resistant plants were regenerated and subsequently transferred to glasshouse conditions. Transformation was confirmed by kanamycin resistance of calli and plants, NPT II ELISA assay and Southern analysis. All transgenic plants were morphologically normal (true-to-type).