Dissolved organic matter (DOM) elucidated from biochars enhances the dissolution of iron oxides and reduction of iron. However, given that reduction mechanism of iron (Fe(III)) in the practical biochar applications for soil amendment and environmental remediation have not been fully elucidated, this study laid great emphasis on the photo-induced Fe(II) liberated from DOM-Fe(III) complexes. Thus, pyrolysis of biomass was carried out at 300 °C to maximize DOM release from biochars. Moreover, three different biomass samples (rice straw (R), granular sludge (G) from an anaerobic digester, and spent coffee grounds (C)) were chosen as carbon substrates for biochars preparation. To demonstrate the transformation of Fe(III), 1 and 5 wt% biochar was applied to the clean (S1) and arsenic-contaminated (S2) soil with/without the light. The results indicate that the light condition produces more Fe(II). The amount of Fe(II) accounts for 25.3, 28.6, and 30.7% of total iron under the light with 5 wt% GB, RB, and CB in S1, and 10.6, 13.1, and 13.8% in S2. This study demonstrates that Fe(II) is generated more under ultraviolet irradiation than visible light and dark condition. In addition, a control experiment without biochar showed that DOM plays an important role in the reduction of Fe(III). The mobility of arsenic increased under the light condition since the intermediates of DOM photo-degradation accelerates the dissolution of iron oxides and arsenic competes with DOM for the adsorption. Therefore, there was no significant correlation between the elution of arsenic and the formation of Fe(II) during the reductive dissolution of iron oxide under the light condition.