Cities have important roles in biodiversity restoration and management. Local municipalities, as key land managers, play critical roles since some species are restricted to small patches of urban habitat, and some are considered threatened. Evolving approaches to urban nature policy, strategy and implementation demonstrate increasing sophistication and complexity, with approaches transitioning towards the adoption of more complex objectives, that address a wider range of urban nature forms, species and ecosystems. These objectives are increasingly underpinned by urban ecology research, citizen science participation and political championing. This paper adopts a ‘dialogue’-adapted research approach to examine the evolution of the City of Melbourne’s urban nature policies and actions across the decade from 2010 to 2020. We identify the key factors that enabled the endorsement and implementation of this capital city’s Nature in the City strategy, analysing the science-policy interface and governance aspects that underpinned this significant policy transition. Through interviews and workshops with a range of actors involved in the implementation of the City of Melbourne’s urban nature policies and activities over the last 10 years, we identify lessons learned that can be applied in other cities by urban land managers. In doing so, we argue that Melbourne’s focus on biodiversity conservation and opportunities for restoration of species and ecological processes constitute important innovations in urban landscape policy.