Identification of salt-responsive mitogen-activated protein kinase cascade members in the halophyte N. sibirica

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BACKGROUND: Soil salinization severely limits global agricultural productivity. Halophytes such as Nitraria sibirica (N. sibirica) serve as valuable model systems for unraveling the sophisticated mechanisms underlying salt adaptation. Although mitogen-activated protein kinase (MAPK) cascades function as central signaling modules in plant stress responses, their specific architecture and functional organization remain largely unexplored in extremophytes . RESULTS: -S/T motifs in MAPKKs, and subfamily-specific signatures in MAPKKKs. Evolutionary analysis classified these components into highly conserved subfamilies aligned with the established Arabidopsis framework. Transcriptomic profiling, coupled with qRT-PCR validation, uncovered multiple salt-responsive genes that exhibited diverse subcellular localization patterns. Furthermore, by integrating AlphaFold-based structural modeling with yeast two-hybrid assays, we identified a linearly organized MAPK cascade module: NsMAPKKK (NISI07G2543) - NsMAPKK (NISI03G1309) - NsMAPK (NISI09G1183). This signaling module was consistently upregulated under salinity stress, and its physical interactions were demonstrated in vitro. CONCLUSIONS: This study presents a genome-wide identification of the MAPK cascade family in N. sibirica. By delineating a candidate salt-responsive module, our findings provide insights into stress signal transduction in extremophytes and offer potential target candidates for engineering salt tolerance in crops.

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