There is now growing evidence that membrane vesicle trafficking proteins,especially of the superfamily of SNAREs,are critical for cellular signalling in plants.Work from this laboratory first demonstrated that a soluble,inhibitory (dominant-negative) fragment of the SNARE NtSyp121 blocked K+ and Cl- channel responses to the stress-related hormone abscisic acid (ABA),but left open a question about functional impacts on signal intermediates,especially on Ca2+-mediated signalling events.Here,we report one mode of action for the SNARE mediated directly through alterations in Caz+ channel gating and its consequent effects on cytosolic-free [Ca2+] ([Ca2+]i) elevation.We find that expressing the same inhibitory fragment of NtSyp121 blocks ABA-evoked stomatal closure,but only partially suppresses stomatal closure in the presence of the NO donor,SNAP,which promotes [Ca2+]i elevation independently of the plasma membrane Ca2+ channels.Consistent with these observations,Ca2+ channel gating at the plasma membrane is altered by the SNARE fragment in a manner effective in reducing the potential for triggering a rise in [Ca2+]i,and we show directly that its expression in vivo leads to a pronounced suppression of evoked [Ca2+]i transients.These observations offer primary evidence for the functional coupling of the SNARE with Ca2+ channels at the plant cell plasma membrane and,because [Ca2+]i plays a key role in the control of K+ and Cl- channel currents in guard cells,they underscore an important mechanism for SNARE integration with ion channel regulation during stomatal closure.