The mechano-sensitive channels of plants may sense increases in tension induced by mechanical stimuli, such as touch, wind and turgor pressure, and a gravitational stimulus. Recent studies have identified plant homologues of the bacterial mechano-sensitive channel MscS, which is gated by membrane tension and reduces intracellular osmolality by releasing small osmolytes from bacterial cells. However, the physiological roles of these homologues have not yet been clearly elucidated, and only two of them have been shown to be involved in the protection of osmotically stressed plastids in Arabidopsis thaliana. We identified another group of candidates for mechano-sensitive channels in Arabidopsis, named MCA1 and MCA2, whose homologues are exclusively found in plant genomes. MCA1 and MCA2 are composed of 421 and 416 amino acid residues, respectively, share 73% homology in their amino acid sequences, and are not homologous to any known ion channels or transporters. Our structural study revealed that the N-terminal region (one to 173 amino acids) of both proteins was necessary and sufficient for Ca2+ influx activity. Interestingly, this region had one putative transmembrane segment containing an Asp residue whose substitution mutation abolished this activity. Our physiological study suggested that MCA1 expressed at the root tip was required for sensing the hardness of the agar medium or soil. In addition, MCA1 and MCA2 were shown to be responsible for hypo-osmotic shock-induced increases in [Ca2+]cyt. Thus, both proteins appear to be involved in the process of sensing mechanical stresses. We discussed the possible role of both proteins in sensing mechanical and gravitational stimuli.