Aquatic plant biomass like Iris pseudacorus can be used as electron donor to improve denitrification performance in subsurface constructed wetlands. However, the phenomenon that the nitrogen removal rate declined in the terminal stage restricted the utilization of litters. In terms of this problem, this study investigated the performance of the used biomass through alkali treatment on nitrogen removal and analyzed the effect of alkali treatment on the component and structure of biomass and microbial community. The results showed that the alkali-treated biomass could further enhance the nitrogen removal by nearly 15% compared with used ones. The significant damage of cell walls and compact fibers containing cellulose and lignin through alkali treatment mainly resulted in the improvement of carbon release and nitrogen removal. With the addition of alkali-treated biomass, the richness index of microbes was higher compared with other biomass materials. Furthermore, the abundance of denitrification related genera increased and the abundance of genera for nitrification was maintained. Based on these finds, a mode of a more efficient Iris pseudacorus self-consumed subsurface flow constructed wetlands was designed. In this mode, the effluent total nitrogen could be stabilized below 5 mg L-1 for nine months and the weight of litters could be further cut down by 75%. These findings would contribute to efficient utilization of plant biomass for nitrogen removal enhancement and final residue reduction in the wetlands.