Influence of sulphate and other anions on leaf apoplastic pH and stomatal closure in Vicia faba under salinity stress

Salinity-induced stress perceived at plant roots elicits complex systemically orchestrated physiological responses in plants, with ionic imbalances and osmotic pressures playing pivotal roles in modulating stomatal regulation. This study investigates how different anions (Cl – , NO 3– , SO 4 2– , and PO 4 3– ) influence root and leaf apoplastic pH (pH apo ) and stomatal closure in Vicia faba under saline conditions. Using a microscopy-based real-time pH apo imaging approach, we demonstrate that transient alkalinisation in the leaf apoplast is anion-specific and varies with leaf maturity, with Cl – and NO 3– eliciting stronger pH apo increases than SO 4 2– and PO 4 3– . Notably, the root apoplast pH remained unaffected by short-term salinity, highlighting that the alkalinisation is localised to the shoot and likely linked to xylem-translocated ionic effects. Findings reveal that stomatal aperture reductions occurred across treatments regardless of the magnitude of apoplastic alkalinisation. Among the anions studied, SO 4 2– displayed unique regulatory potential, suggesting a role in stomatal behaviour. These results provide new insights into the interplay between ionic stress, apoplastic pH regulation, and stomatal responses, offering potential targets for improving salinity tolerance in crops. • Anion-specific pH changes in leaves under salinity stress are linked to stomatal behaviour. • Sulphate shows regulatory potential in leaf apoplastic pH and guard cell function. • Leaf apoplastic pH responses are influenced by anion type and leaf developmental stage. • Transpirational and ionic factors drive shoot-localised apoplastic alkalinisation. • Salinity-induced increases in leaf apoplastic pH are influenced by anion species and leaf maturity, with evidence suggesting a role for sulphate in regulating stomatal behaviour.