The effects of substrate size on the growth of microphytobenthos Achnanthes sp., Amphora sp., Navicula sp. and Nitzschia sp. were examined using glass beads in order for phytoremediation in the benthic layer of coastal waters. The glass beads used in this study were 0.09～0.15 mm (G.B 1), 0.25～0.50 mm (G.B 2), 0.75～1.00 mm (G.B 3) and 1.25～1.65 mm (G.B 4). No addition of glass bead used as control. The specific growth rate and maximum cell density of four microphytobenthos species were increasing with decreasing size of glass beads. Moreover, the control experiment without added attachment substrates showed the lowest specific growth rate and maximum cell density. Therefore, the suitable attachment substrates for mass culture of microphytobenthos seems to be important in order for phytoremediation using microphytobenthos.