Our aim was to elucidate mechanisms underlying nitrogen (N)-deficiency tolerance in bread wheat (cultivar Ruta), conferred by a wild emmer wheat QTL (WEW
IL99). We hypothesised that the tolerance in IL99 is driven by enhanced N-uptake through modification of root system architecture (RSA) underscored by transcriptome modifications. Severe N-deficiency (0.1 N for 26 days) triggered significantly higher plasticity in IL99 compared to Ruta by modifying 16 RSA traits
nine of which were IL99-specific. The change in root growth in IL99 was collectively characterised by a transition in root orientation from shallow to steep, increased root number and length, and denser networks, enabling nutrient acquisition from a larger volume and deeper soil layers. Gene ontology and KEGG-enrichment analyses highlighted IL99-specific pathways and candidate genes elevated under N-deficiency. This included Jasmonic acid metabolism, a key hormone mediating RSA plasticity (AOS1, TIFY, MTB2, MYC2), and lignification-mediated root strengthening (CYP73A, 4CL). 'N-metabolism' was identified as a main shared pathway to IL99 and Ruta, with enhanced nitrate uptake predominant in IL99 (NRT2.4), while remobilisation was the main strategy in Ruta (NRT2.3). These findings provide novel insights into wheat plasticity response underlying tolerance to N-deficiency and demonstrate the potential of WEW for improving N-uptake under suboptimal conditions.