Separation Tree, Historic eucalypt tree in Royal Botanic Gardens Melbourne, Australia

The Separation Tree is a red river gum located in the Royal Botanic Gardens Melbourne, a key site in Victoria's founding history. The tree originally reached about 24 meters in height with a canopy spread of roughly 27 meters and a trunk circumference of about 3.8 meters.

The red river gum stood on the Tennyson Lawn beside a billabong of the Ornamental Lake and pre-dates European settlement. On 15 November 1850 citizens gathered here to celebrate Governor La Trobe's announcement that Victoria would separate from New South Wales. A plaque marking the centenary of separation was placed at its base in 1951, and in 1982 the tree entered the National Trust of Victoria's Significant Tree Register.

The tree's name comes from the 15 November 1850 gathering, when Melbourne residents met here to hear Governor La Trobe announce Victoria's separation from New South Wales. A plaque marking the centenary of that separation was placed at its base in 1951. The tree was added to the National Trust of Victoria's Significant Tree Register in 1982.

The site is located on accessible pathways within the gardens and can be explored alongside other attractions in the area. Information boards explain the context at the location, even though the original tree no longer stands.

The tree was ringbarked in 2010, when about 90 per cent of its cambial tissue was removed; the remaining tissue was destroyed in a second attack in 2013. After the canopy and parts of the trunk were lopped in 2015, only the main trunk and parts of three scaffold limbs remained on the Tennyson Lawn.