Grain yield is increasingly affected by climate factors such as drought and heat. To develop resilient and high-yielding cultivars, high-throughput phenotyping (HTP) techniques are essential for precise decisions in wheat breeding. The ability of unmanned aerial vehicle (UAV)-based multispectral imaging and ensemble learning methods to increase the accuracy of grain yield prediction in practical breeding work is evaluated in this study. For this, 211 winter wheat genotypes were planted under full and limited irrigation treatments, and multispectral data were collected at heading, flowering, early grain filling (EGF), and mid-grain filling (MGF) stages. Twenty multispectral vegetation indices (VIs) were estimated, and VIs with heritability greater than 0.5 were selected to evaluate the models across the growth stages under both irrigation treatments. A framework for ensemble learning was developed by combining multiple base models such as random forest (RF), support vector machine (SVM), Gaussian process (GP), and ridge regression (RR). The R2 values between VIs and grain yield for individual base models were ranged from 0.468 to 0.580 and 0.537 to 0.598 for grain yield prediction in full and limited irrigation treatments across growth stages, respectively. The prediction results of ensemble models were ranged from 0.491 to 0.616 and 0.560 to 0.616 under full and limited irrigation treatments respectively, and were higher than that of the corresponding base learners. Moreover, the grain yield prediction results were observed high at mid grain filling stage under both full (R2 = 0.625) and limited (R2 = 0.628) irrigation treatments through ensemble learning based stacking of four base learners. Further improvements in ensemble learning models can accelerate the use of UAV-based multispectral data for accurate predictions of complex traits like grain yield in wheat.