Tolerance to multiple abiotic stresses is mediated by interacting CNGC proteins that regulate Ca²⁺ influx and...

Members of the cyclic nucleotide-gated channel (CNGC) proteins are reportedly involved in a variety of biotic and abiotic responses and stomatal movement. However, it is unknown if and how a single member could regulate multiple responses. Here we characterized three closely related CNGC genes in rice, OsCNGC14, OsCNGC15 and OsCNGC16, to determine whether they function in multiple abiotic stresses. The loss-of-function mutants of each of these three genes had reduced calcium ion (Ca²⁺) influx and slower stomatal closure in response to heat, chilling, drought and the stress hormone abscisic acid (ABA). These mutants also had reduced tolerance to heat, chilling and drought compared with the wild-type. Conversely, overexpression of OsCNGC16 led to a more rapid stomatal closure response to stresses and enhanced tolerance to heat, chilling and drought. The tight association of stomatal closure and stress tolerance strongly suggests that tolerance to multiple abiotic stresses conferred by these OsCNGC genes results at least partially from their regulation of stomatal movement. In addition, physical interactions were observed among the three OsCNGC proteins but not with a distantly related CNGC, suggesting the formation of hetero-oligomers among themselves. This study unveils the crucial role of OsCNGC14, 15 and 16 proteins in stomatal response and tolerance to multiple stresses, suggesting a mechanism of tolerance to multiple stresses that involves calcium influx and stomatal movement regulation.