Ante- and post-mortem cellular injury dynamics in hybrid poplar foliage as a function of phytotoxic O(3) dose

After reaching phytotoxic levels during the last century, tropospheric ozone (O(3)) pollution is likely to remain a major concern in the coming decades. Despite similar injury processes, there is astounding interspecific–and sometimes intraspecific–foliar symptom variability, which may be related to spatial and temporal variation in injury dynamics. After characterizing the dynamics of physiological responses and O(3) injury in the foliage of hybrid poplar in an earlier study, here we investigated the dynamics of changes in the cell structure occurring in the mesophyll as a function of O(3) treatment, time, phytotoxic O(3) dose (POD(0)), leaf developmental stage, and mesophyll layer. While the number of Hypersensitive Response-like (HR-like) lesions increased with higher O(3) concentrations and POD(0), especially in older leaves, most structural HR-like markers developed after cell death, independent of the experimental factors. The pace of degenerative Accelerated Cell Senescence (ACS) responses depended closely on the O(3) concentration and POD(0), in interaction with leaf age. Changes in total chlorophyll content, plastoglobuli and chloroplast shape pointed to thylakoid membranes in chloroplasts as being especially sensitive to O(3) stress. Hence, our study demonstrates that early HR-like markers can provide reasonably specific, sensitive and reliable quantitative structural estimates of O(3) stress for e.g. risk assessment studies, especially if they are associated with degenerative and thylakoid-related injury in chloroplasts from mesophyll.