Several studies suggest that singers adapt their vocal production to room acoustics, and virtual reality (VR) has increasingly been used to investigate such interactions under controlled conditions. However, questions remain regarding the ecological validity of virtual acoustic environments for studying musicians’ behavior. While prior research has primarily focused on solo singers, the present study explores the impact of acoustic modality (real vs. virtual) on choral performance. A professional four-singer ensemble performed five different choral pieces across five acoustic conditions. Recordings were conducted both in situ, within different spaces of a church, and under corresponding virtual acoustic simulations using auralization techniques. Acoustic and physiological data were collected using close microphones and electroglottography, while subjective perceptions were assessed through questionnaires. Comparative analyses between real and virtual conditions aim to examine how acoustic modality (real or virtual) influences singers’ musical and physiological adaptations, as well as their subjective perceptions.