Maize (Zea mays L.) is one of the most consumed grains in the world. Within the context of continuous climate change and the reduced availability of arable land, it is urgent to breed new maize varieties and screen for the desired traits, e.g., high yield and strong stress tolerance. Traditional phenotyping methods relying on manual assessment are time-consuming and prone to human errors. Recently, the application of uncrewed aerial vehicles (UAVs) has gained increasing attention in plant phenotyping due to their efficiency in data collection. Moreover, hyperspectral sensors integrated with UAVs can offer data streams with high spectral and spatial resolutions, which are valuable for estimating plant traits. In this study, we collected UAV hyperspectral imagery over a maize breeding field biweekly across the growing season, resulting in 11 data collections in total. Multiple machine learning models were developed to estimate the grain yield and flowering time of the maize breeding lines using the hyperspectral imagery. The performance of the machine learning models and the efficacy of different hyperspectral features were evaluated. The results showed that the models with the multi-temporal imagery outperformed those with imagery from single data collections, and the ridge regression using the full band reflectance achieved the best estimation accuracies, with the correlation coefficients (r) between the estimates and ground truth of 0.54 for grain yield, 0.91 for days to silking, and 0.92 for days to anthesis. In addition, we assessed the estimation performance with data acquired at different growth stages to identify the good periods for the UAV survey. The best estimation results were achieved using the data collected around the tasseling stage (VT) for the grain yield estimation and around the reproductive stages (R1 or R4) for the flowering time estimation. Our results showed that the robust phenotyping framework proposed in this study has great potential to help breeders efficiently estimate key agronomic traits at early growth stages.