Canopy cover is a key forest structural parameter that is commonly used in forest inventory, sustainable forest management and maintaining ecosystem services. Recently, much attention has been paid to the use of unmanned aerial vehicle (UAV)-based light detection and ranging (LiDAR) due to the flexibility, convenience, and high point density advantages of this method. In this study, we used UAV-based LiDAR data with individual tree segmentation-based method (ITSM), canopy height model-based method (CHMM), and a statistical model method (SMM) with LiDAR metrics to estimate the canopy cover of a pure ginkgo (Ginkgo biloba L.) planted forest in China. First, each individual tree within the plot was segmented using watershed, polynomial fitting, individual tree crown segmentation (ITCS) and point cloud segmentation (PCS) algorithms, and the canopy cover was calculated using the segmented individual tree crown (ITSM). Second, the CHM-based method, which was based on the CHM height threshold, was used to estimate the canopy cover in each plot. Third, the canopy cover was estimated using the multiple linear regression (MLR) model and assessed by leave-one-out cross validation. Finally, the performance of three canopy cover estimation methods was evaluated and compared by the canopy cover from the field data. The results demonstrated that, the PCS algorithm had the highest accuracy (F = 0.83), followed by the ITCS (F = 0.82) and watershed (F = 0.79) algorithms; the polynomial fitting algorithm had the lowest accuracy (F = 0.77). In the sensitivity analysis, the three CHM-based algorithms (i.e., watershed, polynomial fitting and ITCS) had the highest accuracy when the CHM resolution was 0.5 m, and the PCS algorithm had the highest accuracy when the distance threshold was 2 m. In addition, the ITSM had the highest accuracy in estimation of canopy cover (R2 = 0.92, rRMSE = 3.5%), followed by the CHMM (R2 = 0.94, rRMSE = 5.4%), and the SMM had a relative low accuracy (R2 = 0.80, rRMSE = 5.9%).The UAV-based LiDAR data can be effectively used in individual tree crown segmentation and canopy cover estimation at plot-level, and CC estimation methods can provide references for forest inventory, sustainable management and ecosystem assessment.