Nothofagus pumilio is the most important forest species in South Patagonia. The present work was carried out to determine the suitable cultural conditions for in vitro propagation through shoot culture of saplings and a mature tree. The days after sprouting and the presence of the bud scales were critical for culture initiation. A medium with a low salt concentration and the addition of glutamine, as Broadleaved Tree Medium, was suitable for shoot multiplication, being affected both by cytokinin and giberellin. A multiplication rate of 1:3-5 was obtained with 20­ 30 μM 2iP and 0.3 μM GA3 at day 63. No significant differences were found between the juvenile and adult material. Rooted shoots (60-70%) were obtained with 0.61-1.23 μM AIB and ten days darkness at the induction stage. Rooted shoots were then successfully acclimatised and 43.2% of good quality plants were obtained after 45 days.