Decoding Salinity Tolerance in Salicornia europaea L.: Image-Based Oxidative Phenotyping and Histochemical Mapping of...

Halophytes such as Salicornia europaea rely on biochemical and structural mechanisms to survive in saline environments. This study aimed to evaluate oxidative stress and structural defense responses in four inland populations-Poland (Inowrocław, Ciechocinek), Germany (Salzgraben-Salzdahlum, Salz), and Soltauquelle (Soltq)-subjected to 0, 200, 400, and 1000 mM NaCl, using non-destructive, image-based approaches. Lipid peroxidation was assessed via malondialdehyde (MDA) detected with Schiff's reagent, and hydrogen peroxide (H₂O₂) accumulation was visualized with 3,3'-diaminobenzidine (DAB). Roots and shoots were analyzed through colour image analysis and quantified using a computer vision system (CVS). MDA accumulation revealed population-specific differences, with Salz tending to exhibit lower peroxidation, characterized by lower L* ≈ 42-43 and higher b* ≈ 37-18 in shoots at 200-400 mM, which may reflect a potentially more effective salt-management strategy. Although H₂O₂ responses deviated from a direct salinity-dependent trend, particularly in the tolerant Salz and Soltq populations, both approaches effectively tracked population-specific adaptation, with German populations displaying detectable basal H₂O₂ levels, consistent with its multifunctional signalling role in salt management and growth regulation. Structural defences were further explored through histochemical mapping and image analysis of pectin and lignin distribution, which revealed population-specific patterns consistent with cell wall remodelling under stress. Non-destructive, image-based methods proved effective for detecting oxidative and structural responses in halophytes. Such a non-destructive, cost-efficient, and reproducible approach can accelerate the identification of salt-tolerant ecotypes for saline agriculture and reinforce S. europaea as a model species for elucidating salt-tolerance mechanisms.