Oxygen defects of nanoflower TiO2 photo-catalyst was fabricated at the presence of hydrogen at different temperatures (100–600∘C) and the concentrations of oxygen defects were firstly quantitatively analyzed by hydrogen programmed temperature reduction techniques (H2-TPR). Total oxygen defect concentration and surface oxygen defect concentration were consistent with XPS and EPR results, respectively. Even at the hydrogen thermal temperature of 600∘C, the shape of TiO2 was still kept as nanoflower structure as characterized by SEM. However, the rutile and anatase coexist in the composition of crystal phase when hydrogen reduction temperature of the TiO2 catalyst reached 400∘C to 600∘C as proved by Raman and XRD results. TiO2 sample with oxygen defects shows excellent photo-catalytic activity for degradation of Direct Blue 78(DB) regardless of ultraviolet light (the maximum degradation rate achieved within 100[Formula: see text]min was 93.27%) or visible light (the maximum degradation rate achieved within 100[Formula: see text]min was 88.25%). The photo-catalytic activity seems to be highly correlated with the surface oxygen defects of TiO2 catalyst. With surface oxygen-defect concentrations increase, the degradation ability on DB was significantly enhanced, while bulk oxygen defects had negligible effect on the photo-catalytic activity. The enhanced photo-catalytic performance of TiO2 with a fixed amount of oxygen defects was attributed to the strong capturing capability of the photo-generated electrons. In addition, the surface defects could also improve the photo-catalytic reaction efficiency.