Drought is a major abiotic stress that limits wheat production worldwide. To ensure food security for the rapidly increasing world population, improving wheat yield under drought stress is urgent and relevant. In this study, an RNA-seq analysis was conducted to study the effect of drought on wheat transcriptome changes during reproductive stages under field conditions. Our results indicated that drought stress during early reproductive periods had a more severe impact on wheat development, gene expression and yield than drought stress during flowering. In total, 115,656 wheat genes were detected, including 309 differentially expressed genes (DEGs) which responded to drought at various developmental stages. These DEGs were involved in many critical processes including floral development, photosynthetic activity and stomatal movement. At early developmental stages, the proteins of drought-responsive DEGs were mainly located in the nucleus, peroxisome, mitochondria, plasma membrane and chloroplast, indicating that these organelles play critical roles in drought tolerance in wheat. Furthermore, the validation of five DEGs confirmed their responsiveness to drought under different genetic backgrounds. Functional verification of DEGs of interest will occur in our subsequent research. Collectively, the results of this study not only advanced our understanding of wheat transcriptome changes under drought stress during early reproductive stages but also provided useful targets to manipulate drought tolerance in wheat at different development stages.