Reserved as parkland in 1854 by Superintendent La Trobe after nearly 14 years of petitioning, Royal Park initially occupied an area of some 700 acres (280 ha), including the site of present-day Princes Park and the Melbourne General Cemetery, although its size gradually diminished as portions were excised for development. In 1858 the north-west corner became an experimental farm. Before 1862 sheep were the main occupiers of the parkland, and local citizens complained of its neglected and overgrown state. In 1861 the Zoological Gardens were given 50 acres (20 ha), with an additional 500 acres (200 ha) granted temporarily for grazing animals. In 1861 explorers Burke and Wills set off on their expedition across Australia from Royal Park, and a fenced tree marked their departure point, later replaced by a cairn (unveiled in 1890).
In the 1870s portions of the park were cleared, but poor soil, harsh conditions and insufficient funds hampered the park's improvement. In 1876, 424 acres (170 ha) were permanently reserved as parkland, excluding the zoo, agricultural showground and powder magazine located within its grounds. Royal Park Golf Course, established in 1903, was the first of the park's sporting facilities, which today include a hockey and netball centre and numerous ovals.
During the world wars, Royal Park was a temporary Army camp, and from 1946 to 1956 an emergency housing site. Today the vegetation is predominantly grassland interspersed with woodlands, and its sparsity is somewhat unusual for a public park in Melbourne. Native trees, such as red and sugar gums, generally planted from the 1930s, when nationalistic sentiments and the desire for Australian flora were high, are plentiful. The Royal Park Protection Group now endeavours to 'protect, regenerate and conserve' the park and oppose actions by government, commercial or sporting bodies that restrict public access to Royal Park.