Arsenic (As) hyperaccumulator Pteris ferns are renowned for their capacity to accumulate As and have been used to remediate As-contaminated environmental. However, there is less information on how they perform under low temperature though it is important for practical phytoremediation. The purpose of this study was to identify the effect of temperature on As accumulation by three As hyperaccumulators, Pteris multifida, Pteris cretica and Pteris vittata. Ferns were cultured with 5 mg/L As addition under 25 °C to 5 °C for 15 days. The results showed that dropping of temperatures reduced As accumulation by P. vittata moderately but not P. multifida and P. cretica until 10 °C. At 5 °C, all ferns discontinued As accumulation, and the morphology showed necrosis in P. vittata, wherein P. multifida and P. cretica kept healthy. The As distribution showed that As was mainly accumulated in fronds, while P. multifida stored partial As in its root. Both translocation factor and As efflux showed that temperate zone ferns manage As more strictly as compared to P. vittata. Our findings demonstrated that temperature should be considered when applying Pteris ferns for As phytoremediation, and P. multifida could be the most suitable fern for treating As-contaminated water in temperate zone area.