Hijacked hydraulics: <i>Verticillium dahliae</i> ‐induced xylem dysfunction in pepper stems revealed by integrated hydraulic, imaging, and molecular analyses

Xylem tissue enables efficient long-distance water transport but is a primary target for vascular pathogens. This study investigates how systemic invasion by Verticillium dahliae impairs the hydraulic function of pepper (Capsicum annuum) plants, focussing on xylem colonisation and its anatomical and physiological effects. Real-time sap flow was continuously monitored with custom-built ExoBeat sensors, while periodic stem water potential measurements allowed calculation of changes in stem hydraulic conductance as an additional indicator of xylem performance. Fungal colonisation was assessed by quantitative polymerase chain reaction, and vessel occlusions and embolised conduits were visualised using scanning electron microscopy and micro-computed tomography, complemented by direct hydraulic conductivity measurements. By 14 d post inoculation, V. dahliae had progressed from roots to aboveground tissues, coinciding with a marked decrease in sap flow, water potential, and soil-to-stem hydraulic conductance, alongside the onset of dwarfing. Direct fungal blockage and anatomical changes were the primary contributors to hydraulic dysfunction. Vessel occlusion by tyloses, gels, and air embolisms played a negligible role. This study reveals how V. dahliae progressively impairs pepper hydraulics through systemic xylem colonisation, highlighting the value of real-time sap flow monitoring. Our integrative, multidisciplinary approach offers a powerful framework to unravel the complexity of dynamic plant-fungal vascular interactions.