Alumina-forming austenitic (AFA) steel exhibits excellent high-temperature properties and has attracted significant attention in thermal power generation industry. Microstructure evolutions and yield strengths of 20Ni-AFA steel during isothermal aging are investigated. The results show that five types of phases are found in 20Ni-AFA steel during isothermal ageing: NbC, L12-Ni3Al, M23C6, B2-NiAl and Laves phases. Addition of Cu causes precipitation of L12-Ni3Al phase in 20Ni-AFA steel at the early stage of isothermal aging. L12-Ni3Al phase grows and gradually transforms into B2-NiAl phase which possesses a more stable structure with the extension in isothermal aging time. The transformation leads to yield strengths of 20Ni-AFA steel to decrease anomalously during 24-120 h isothermal aging. Yield strengths of 20Ni-AFA steel increase again as isothermal aging time continues to increase.