Eucalyptus spp. is a dominant tree genus in Australia and most Eucalyptus spp. are canopy dominant species. In Australian natural forests, Eucalyptus spp. commonly are associated with understorey legumes which play a crucial role for ecological restoration owing to their nitrogen (N) fixing ability for replenishing the soil N lost after frequent prescribed burning. This study aimed to explore to what extent physiological responses of these species differ 7 and 12 years after last fire. Two most common understorey Acacia spp., Acacia leiocalyx and A. disparrima, as well as one non-leguminous Eucalyptus resinifera, were studied due to their dominance in the forest. Both A. leiocalyx and A. disparrima showed higher carbon (C) assimilation capacity, maximum photosynthetic capacity, and moderate foliar C/N ratio compared with E. resinifera. A. leiocalyx showed various advantages compared to A. disparrima such as higher photosynthetic capacity, adaptation to wider light range and higher foliar total N (TNmass). A. leiocalyx also relied on N2-fixing ability for longer time compared to A. disparrima. The results suggested that the two Acacia spp. were more beneficial to C and N cycles for the post burning ecosystem than the non-N2-fixing species E. resinifera. A. leiocalyx had greater contribution to complementing soil N cycle long after burning compared to A. disparrima.