We investigated a nodulin 26-like protein NIP2;1, which belongs to the third subgroup of Arabidopsis aquaporins. Histochemical analysis of a promoter-beta-glucuronidase fusion revealed the root-specific expression of NIP2;1. The NIP2;1 protein was detected in young roots, but not in leaves, stems, flowers or siliques. The transient expression of NIP2;1 linked with green fluorescent protein in Arabidopsis cultured cells showed its putative endoplasmic reticulum (ER) localization. NIP2;1 expressed in yeast cells had low water channel activity in the membranes. NIP2;1 may function as a water channel and/or ER channel for other small molecules or ions.