Genome-Wide analysis of the NRAMP gene family in Arabidopsis thaliana: identification, expression and response to multiple heavy metal stresses and phytohormones

Heavy metal stress is a critical challenge to agricultural productivity, necessitating deeper insights into the molecular mechanisms of metal transport in plants. In this study, we conducted a comprehensive genome-wide characterization of the Natural Resistance-Associated Macrophage Protein (NRAMP) gene family in Arabidopsis thaliana and identified six AtNRAMP genes. Phylogenetic and synteny analyses revealed their distribution into two distinct clades and evolutionary conservation with legumes such as Glycine max and Arachis hypogaea, indicating functional divergence and gene duplication events maintained under purifying selection. Conserved protein motifs and domains, particularly the NRAMP transmembrane domain, highlighted their conserved role in divalent metal ion transport, while cis-regulatory element analysis demonstrated enrichment of stress- and hormone-responsive elements, pointing to tight transcriptional regulation under environmental challenges. Structural modeling further supported the functional conservation of AtNRAMP proteins. Expression profiling showed clear tissue-specific expression under normal conditions and strong, differential regulation in response to cadmium and other heavy metals, as well as to the phytohormone abscisic acid (ABA). Collectively, these results provide foundational insights into the evolutionary relationships, regulatory mechanisms, and stress-responsive expression of the AtNRAMP gene family, offering a framework for future functional studies and potential applications in developing crops with enhanced heavy metal tolerance and improved growth under stress conditions.