AbstractEcosystem respiration (Reco) arises from interacting autotrophic and heterotrophic processes constrained by distinct drivers. Here, we evaluated up‐scaling of observed components of Reco in a mature eucalypt forest in southeast Australia and assessed whether a land surface model adequately represented all the fluxes and their seasonal temperature responses. We measured respiration from soil (Rsoil), heterotrophic soil microbes (Rh), roots (Rroot), and stems (Rstem) in 2018–2019. Reco and its components were simulated using the CABLE–POP (Community Atmosphere‐Biosphere Land Exchange–Population Orders Physiology) land surface model, constrained by eddy covariance and chamber measurements and enabled with a newly implemented Dual Arrhenius and Michaelis‐Menten (DAMM) module for soil organic matter decomposition. Eddy‐covariance based Reco (Reco.eddy, 1,439 g C m−2 y−1) was slightly higher than the sum of the respiration components (Reco.sum, 1,295 g C m−2 y−1) and simulated Reco (1,297 g C m−2 y−1). The largest mean contribution to Reco was from Rsoil (64%) across seasons. The measured contributions of Rh (49%), Rroot (15%), and Rstem (22%) to Reco.sum were very close to model outputs of 46%, 11%, and 22%, respectively. The modeled Rh was highly correlated with measured Rh (R2 = 0.66, RMSE = 0.61), empirically validating the DAMM module. The apparent temperature sensitivities (Q10) of Reco were 2.22 for Reco.sum, 2.15 for Reco.eddy, and 1.57 for CABLE‐POP. This research demonstrated that bottom‐up respiration component measurements can be successfully scaled to eddy covariance‐based Reco and help to better constrain the magnitude of individual respiration components as well as their temperature sensitivities in land surface models.