Two cold-tolerant species (Eucalyptus macarthurii Deane et Maiden and E. smithii R.T. Baker), a cold-tolerant hybrid (E. macarthurii×E. grandis Hill ex Maiden), and E. saligna Sm. were propagated in vitro from nodal explants collected from field-grown seedlings and from clonal hedges. Shoot growth was initiated on modified Murashige and Skoog (MS) medium containing BA at 0.1 mg·liter-1. Modified MS medium with BA (0.2 mg·liter-1) and NAA (0.01 mg·liter-1) was most effective in promoting shoot proliferation. Root initiation was achieved on half-strength modified MS medium with 2 mg IBA/liter. Rooted plants were hardened and established in the field. Chemical names used: N-(phenylmethyl)-1EZ-purin-6-amine (BA); 2-(1-naphthyl)acetic acid (NAA); 1H-indole-3-butyric acid (IBA).