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Background  Mechanosensing and its downstream responses are speculated to involve sensory complexes containing Ca 2+ -permeable mechanosensitive channels. On recognizing osmotic signals, plant cells initiate activation of a widespread signal transduction network that induces second messengers and triggers inducible defense responses. Characteristic early signaling events include Ca 2+  influx, protein phosphorylation and generation of reactive oxygen species (ROS). Pharmacological analyses show Ca 2+  influx mediated by mechanosensitive Ca 2+  channels to influence induction of osmotic signals, including ROS generation. However, molecular bases and regulatory mechanisms for early osmotic signaling events remain poorly elucidated.    Results  We here identified and investigated OsMCA1, the sole rice homolog of putative Ca 2+ -permeable mechanosensitive channels in Arabidopsis (MCAs). OsMCA1 was specifically localized at the plasma membrane. A promoter-reporter assay suggested that  OsMCA1  mRNA is widely expressed in seed embryos, proximal and apical regions of shoots, and mesophyll cells of leaves and roots in rice. Ca 2+  uptake was enhanced in  OsMCA1 -overexpressing suspension-cultured cells, suggesting that OsMCA1 is involved in Ca 2+  influx across the plasma membrane. Hypo-osmotic shock-induced ROS generation mediated by NADPH oxidases was also enhanced in  OsMCA1- overexpressing cells. We also generated and characterized  OsMCA1-RNAi  transgenic plants and cultured cells;  OsMCA1- suppressed plants showed retarded growth and shortened rachises, while  OsMCA1 -suppressed cells carrying Ca 2+ -sensitive photoprotein aequorin showed partially impaired changes in cytosolic free Ca 2+  concentration ([Ca 2+ ] cyt ) induced by hypo-osmotic shock and trinitrophenol, an activator of mechanosensitive channels.    Conclusions  We have identified a sole MCA ortholog in the rice genome and developed both overexpression and suppression lines. Analyses of cultured cells with altered levels of this putative Ca 2+ -permeable mechanosensitive channel indicate that OsMCA1 is involved in regulation of plasma membrane Ca 2+  influx and ROS generation induced by hypo-osmotic stress in cultured rice cells. These findings shed light on our understanding of mechanical sensing pathways.

bmc plant biology

Plasma membrane protein OsMCA1 is involved in regulation of hypo-osmotic shock-induced Ca2+influx and modulates generation of reactive oxygen species in cultured rice cells