Identification of earliness per se (Eps) flowering time loci in spring wheat are troublesome due to confounding effects of vernalization and photoperiod responses. The Wheat Association Mapping Initiative panel of 287 elite lines was assessed to identify genomic regions associated with Eps and to understand the effects of vernalization and photoperiod treatments in spring wheat. The panel was grown under field conditions with four different treatments: (i) vernalization, where 1-d germinated seeds were kept at 4°C for 6 to 8 wk; (ii) extended photoperiod treatment, from seedling emergence to 10 d after anthesis in the field; (iii) treatments (i) and (ii) in combination; and (iv) a control treatment without either (i) or (ii). The combined vernalization and photoperiod treatments had the greatest effect in advancing the flowering time (4 d). Genome-wide association study in this panel on treatment (iii) detected a locus for Eps in chromosome 1D at 163 to 167 cM. An analysis of the flowering time data on this panel from 19 international environments did not show the presence of Eps in chromosome 1D, indicating the possible masking effect of vernalization and photoperiod genes. This study also validated the diagnostic marker—a transcription factor in the circadian clock input pathway called Triticum aestivum EARLY FLOWERING 3—that is associated with Eps in chromosome 1D and can be used for marker-assisted selection.