Probing ozone effects on European hornbeam (Carpinus betulus L. and Ostrya carpinifolia Scop.) leaf water content through THz imaging and dynamic stomatal response

We investigated the impact of ozone exposure on Hornbeam using a novel dual approach based on Terahertz (THz) imaging in a free-air ozone exposure experiment (three ozone levels: ambient; 1.5 times ambient; twice ambient). The research aims at unraveling the physiological responses induced by elevated ozone levels on water dynamics. THz imaging unveiled dynamic changes in leaf water content, providing a non-invasive approach to leaf water monitoring. Leaf gas exchange measurements assessed stomatal responses to light variation. Our findings showcase a compelling correlation between elevated ozone levels and reduction in photosynthetic rate and impairment of stomatal function, i.e. "stomatal sluggishness", indicative of nuanced regulatory mechanism. Stomatal sluggishness was particularly evident in Carpinus betulus (CB) compared to Ostrya carpinifolia (OC) and was linked to reduction in photosynthetic capacity. THz-based imaging techniques confirmed this result indicating a negative effect of O3 on leaf-level total water content. In addition, spatial analysis of leaf water status using these techniques also highlighted that the negative effect of O3 on water status was progressing even in less sensitive OC plants though visible foliar injury was not detected. In fact, OC showed a relative dry area of 1.6 ± 1.6 % in the control group and 3.8 ± 1.3 % under high ozone levels. THz-based imaging techniques provided a deep understanding of O3 behavior in plants and may be recommended for precision biosensing in the early detection of O3-induced damage. The integration of THz imaging and physiological analysis resulted in comprehensive understanding of Hornbeam acclimation response to ozone exposure.