As an important forest type, deciduous broad-leaved forest is crucial for estimating forest carbon sequestration capacity and evaluating forest carbon balance. This study focuses on the natural deciduous broad-leaved forest of Mazongling Nature Reserve in Jinzhai County of China. WorldView-2 images were selected as data source. 36 candidate factors including vegetation indices, texture features, and topographic factors were used for modelling. Three machine learning algorithms (i.e., random forest, k-nearest neighbor, and artificial neural network) were used to establish the optimal quantitative retrieval model for natural deciduous broad-leaved biomass. Results showed that the ANN model was the best predictor with R 2  = 0.69 and RMSE = 31.53 (Mg·ha −1 ). Combining the ANN model with the complete spatial coverage of remote sensing data, we developed a distribution map of natural deciduous broad-leaved biomass in the Mazongling forest farm. The estimated average biomass of the study area was 90.34 ± 47.96 Mg·ha −1 . In addition, the influence of light saturation on model accuracy is also discussed. This study confirms that remote sensing data in temporal and spatial space can improve the model estimation accuracy.