A series of Sr0.95Al2O4:Eu2+0.02, Dy3+ 0.03•nB2O3 (0≤n≤0.30) were prepared by a solid state reaction. The role of B2O3 on the photoluminescence (PL) properties of Sr0.95Al2O4:Eu2+0.02, Dy3+0.03 were evaluated with UV emission spectra and decay curves. B2O3 significantly improved the emission intensity and persistent luminescence time. The crystal environment and defects induced by B3+ doping were detected with IR spectra, thermoluminescence and positron annihilation (PA) methods. Some Al3+ were substituted by B3+ and a kind of defect complex cluster responsible for the long decay time was formed when the substitution of Al3+ by B3+ occurs.