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Agrobacterium-mediated genetic transformation system was established in Dendrobium Formidible ‘Ugusu’ by inoculating PLBs with A. tumefaciens strain EHA101 (pIG121Hm) harboring hygromycin phosphotranferase (hpt) and neomycin phosphotranferase II (nptII) genes as selectable marker gene and β-glucuronidase (gus) gene as a reporter gene. For obtaining the optimum conditions for the transformation, several factors such as the stage of PLBs after subculture, bacterial concentrations, kind of inoculation medium, inoculation time and inoculation condition (with or without rotary shaking), were examined. After inoculation, PLBs were cocultivated for 3 days and then transferred for selection onto 2.5 g l−1 gellan gum-solidified ND medium containing 10 g l−1 maltose, 20 mg l−1 hygromycin and 20 mg l−1 meropenem. Hygromycin-resistant plantlets were regenerated from secondary PLBs after 4 months of selection. Transformation of these plants was confirmed by GUS histochemical assay, PCR and Southern blot analyses. The highest transformation efficiency of 18.5% was obtained when PLBs 3 weeks after subculture were inoculated with 1 : 10 diluted bacteria (OD600≈0.1) with liquid medium containing only 10 g l−1 maltose and 100 μM acetosyringone with shaking for 30 min.

<i>Agrobacterium</i>-mediated transformation of protocorm-like bodies in <i>Dendrobium</i> Formidible ‘Ugusu’