Forest spatial structure is of significant importance for studying forest biomass accumulation and management. However, above-ground biomass (AGB) estimation based on satellite remote sensing struggles to capture forest spatial structure information, which to some extent affects the accuracy of AGB estimation. To address this issue, this study focused on Chinese fir (Cunninghamia lanceolata) plantations in Zhejiang Province. Using UAV-LiDAR (unmanned aerial vehicle light detection and ranging) data and a seed-point-based individual tree segmentation algorithm, information on individual fir trees was obtained. Building on this foundation, structural parameters such as neighborhood comparison (U), crowding degree (C), uniform angle index (W), competition index (CI), and canopy openness (K) were calculated, and their distribution characteristics analyzed. Finally, these parameters were integrated with UAV-LiDAR point cloud features to build machine learning models, and a geographical detector was used to quantify their contribution to AGB estimation. The research findings indicate the following: (1) The studied stands exhibited a random spatial pattern, moderate competition, and sufficient growing space. (2) A significant correlation existed between the U and AGB (r > 0.6), followed by CI. The optimal stand structure for AGB accumulation was C = 0.25, U < 0.5, CI in (0, 0.8], and K > 0.3. (3) The four machine learning models constructed by coupling spatial structure with point cloud features all improved the accuracy of AGB estimation for the fir forest to some extent. Among them, the XGBoost model performed best, achieving a model accuracy (R²) of 0.92 and a relatively low error (RMSE = 14.02 kg). (4) Geographical detector analysis indicated that U and CI contributed most to AGB estimation, with q-values of 0.44 and 0.37, respectively.