Physiological maturity date is a critical parameter for the selection of breeding lines in soybean breeding programs. The conventional method to estimate the maturity dates of breeding lines uses visual ratings based on pod senescence by experts, which is subjective by human estimation, labor-intensive and time-consuming. Unmanned aerial vehicle (UAV)-based phenotyping systems provide a high-throughput and powerful tool of capturing crop traits using remote sensing, image processing and machine learning technologies. The goal of this study was to investigate the potential of predicting maturity dates of soybean breeding lines using UAV-based multispectral imagery. Maturity dates of 326 soybean breeding lines were taken using visual ratings from the beginning maturity stage (R7) to full maturity stage (R8), and the aerial multispectral images were taken during this period on 27 August, 14 September and 27 September, 2018. One hundred and thirty features were extracted from the five-band multispectral images. The maturity dates of the soybean lines were predicted and evaluated using partial least square regression (PLSR) models with 10-fold cross-validation. Twenty image features with importance to the estimation were selected and their changing rates between each two of the data collection days were calculated. The best prediction (R² = 0.81, RMSE = 1.4 days) was made by the PLSR model using image features taken on 14 September and their changing rates between 14 September and 27 September with five components, leading to the conclusion that the UAV-based multispectral imagery is promising and practical in estimating maturity dates of soybean breeding lines.