Crosstalk between melatonin and Ca²⁺/CaM evokes systemic salt tolerance in Dracocephalum kotschyi

In this study, the role of calcium/calmodulin (Ca²⁺/CaM) and melatonin (Mel) as two signal molecules in inducing systemic salt tolerance of Dracocephalum kotschyi Boiss. was investigated. Salinity stress (100 mM NaCl) reduced plant growth and induced ionic, osmotic, and oxidative damages in D. kotschyi leaves. Detection of cytosolic free Ca²⁺ ([Ca²⁺]_cyt) by the Fura-2 method and the measurement of endogenous Mel by GC-MS demonstrated that salinity induced Ca²⁺ burst and increased endogenous Mel content in D. kotschyi leaves. Root pretreatment with 5 mM Ca²⁺ or 100 μM Mel recovered plant growth, reduced leaf electrolytic leakage, H₂O_2, and MDA contents and improved membrane integrity not only at the application site (roots), but also at the untreated distal parts (leaves) under salt stress. Rhizospheric treatment with Mel and Ca²⁺ triggered systemic tolerance in D. kotschyi, as judged from improving RWC, increasing proline content, modulating Na⁺, K⁺, and Ca²⁺ homeostasis, and enhancing the activities of SOD, CAT, APX, and POD in the leaves of salt-stressed plants. Mel augmented [Ca²⁺]_cyt, but the rhizospheric application of Ca²⁺ antagonists impaired the latter responses. Furthermore, root pretreatment with Ca²⁺ increased Mel content, but the application of p-chlorophenylalanine (as an inhibitor of Mel biosynthesis) decreased the above attributes in the leaves of Ca²⁺-treated plants, leading to an arrest in the Ca²⁺-induced systemic salt tolerance. These novel results suggest that interaction of Ca²⁺/CaM and Mel is involved in overcoming salt-induced ionic, osmotic, and oxidative damages and Ca²⁺ and Mel may act as long-distance signals for inducing systemic salt tolerance in D. kotschyi.