<i>Arabidopsis</i> CaM1 and CaM4 Promote Nitric Oxide Production and Salt Resistance by Inhibiting <i>S</i>-Nitrosoglutathione Reductase via Direct Binding

<div><p>Salt is a major threat to plant growth and crop productivity. Calmodulin (CaM), the most important multifunctional Ca<sup>2+</sup> sensor protein in plants, mediates reactions against environmental stresses through target proteins; however, direct proof of the participation of CaM in salt tolerance and its corresponding signaling pathway <i>in vivo</i> is lacking. In this study, we found that <i>AtCaM1</i> and <i>AtCaM4</i> produced salt-responsive <i>CaM</i> isoforms according to real-time reverse transcription-polymerase chain reaction analyses; this result was verified based on a phenotypic analysis of salt-treated loss-of-function mutant and transgenic plants. We also found that the level of nitric oxide (NO), an important salt-responsive signaling molecule, varied in response to salt treatment depending on <i>AtCaM1</i> and <i>AtCaM4</i> expression. GSNOR is considered as an important and widely utilized regulatory component of NO homeostasis in plant resistance protein signaling networks. <i>In vivo</i> and <i>in vitro</i> protein-protein interaction assays revealed direct binding between AtCaM4 and <i>S</i>-nitrosoglutathione reductase (GSNOR), leading to reduced GSNOR activity and an increased NO level. Overexpression of <i>GSNOR</i> intensified the salt sensitivity of <i>cam4</i> mutant plants accompanied by a reduced internal NO level, whereas a <i>gsnor</i> deficiency increased the salt tolerance of <i>cam4</i> plants accompanied by an increased internal NO level. Physiological experiments showed that CaM4-GSNOR, acting through NO, reestablished the ion balance to increase plant resistance to salt stress. Together, these data suggest that AtCaM1 and AtCaM4 serve as signals in plant salt resistance by promoting NO accumulation through the binding and inhibition of GSNOR. This could be a conserved defensive signaling pathway in plants and animals.</p></div>