Node-derived shoot cultures of Eucalyptus regnans were established from in vitro grown seedlings on Murashige and Skoog basal medium supplemented with 0.5 mg L-1 (2 µm) zeatin and 0.05 mg L-1 (0.3 µm) napthaleneacetic acid. A double sterilisation method was essential to obtain clean material from seed. Microcuttings from established cultures were used to develop an efficient method for in vitro rooting. Rooting was best after a 7 day pulse on 20 mg L-1 (98 µm) indolebutyric acid. Hoagland's or Woody Plant Medium supported better rooting than MS basal medium and rooting was significantly enhanced by subculture to activated charcoal after the auxin pulse. Carbohydrate (sucrose or glucose) was essential for rooting while high light intensity was inhibitory. Optimal light conditions were a 12 h day (17 W m-2). In all, 90% of plantlets established in the nursery survived the winter.