The impact of ozone (O3) pollution events on the plant drought response needs special attention because spring O3 episodes are often followed by summer drought. By causing stomatal sluggishness, O3 could affect the stomatal dynamic during a subsequent drought event. In this context, we studied the impact of O3 exposure and water deficit (in the presence or in the absence of O3 episode) on the stomatal closure/opening mechanisms relative to irradiance or vapour pressure deficit (VPD) variation. Two genotypes of Populus nigra x deltoides were exposed to various treatments for 21 days. Saplings were exposed to 80 ppb/day O3 for 13 days, and then to moderate drought for 7 days. The curves of the stomatal response to irradiance and VPD changes were determined after 13 days of O3 exposure, and after 21 days in the case of subsequent water deficit, and then fitted using a sigmoidal model. The main responses under O3 exposure were stomatal closure and sluggishness, but the two genotypes showed contrasting responses. During stomatal closure induced by a change in irradiance, closure was slower for both genotypes. Nonetheless, the genotypes differed in stomatal opening under light. Carpaccio stomata opened more slowly than control stomata, whereas Robusta stomata tended to open faster. These effects could be of particular interest, as stomatal impairment was still present after O3 exposure and could result from imperfect recovery. Under water deficit alone, we observed slower stomatal closure in response to VPD and irradiance, but faster stomatal opening in response to irradiance, more marked in Carpaccio. Under the combined treatment, most of the parameters showed antagonistic responses. Our results highlight that it is important to take genotype-specific responses and interactive stress cross-talk into account to improve the prediction of stomatal conductance in response to various environmental modifications.

Application of nanopesticides may substantially increase surface attachment and internalization of engineered nanoparticles (ENPs) in food crops. This study investigated the role of stomata in the internalization of silver nanoparticles (Ag NPs) using abscisic acid (ABA)-responsive ecotypes (Ler and Col-7) and ABA-insensitive mutants (ost1-2 and scord7) of Arabidopsis thaliana in batch sorption experiments, in combination with microscopic visualization. Compared with those of the ABA-free control, stomatal apertures were significantly smaller for the Ler and Col-7 ecotypes (p ˂ 0.05) but remained unchanged for the ost1-2 and scord7 mutants, after exposure to 10 μM ABA for 1 h. Generally Ag NP sorption to the leaves of the Ler and Col-7 ecotypes treated with 10 μM ABA was lower than that in the ABA-free control, mainly due to ABA-induced stomatal closure. The difference in Ag NP sorption with and without ABA was less pronounced for Col-7 than for Ler, suggesting different sorption behaviors between these two ecotypes. In contrast, there was no significant difference in foliar sorption of Ag NPs by the ost1-2 and scord7 mutants with and without ABA treatment. Ag NPs were widely attached to the Arabidopsis leaf surface, and found at cell membrane, cytoplasm, and plasmodesmata, as revealed by scanning electron microscopy and transmission electron microscopy, respectively. These results highlight the important role of stomata in the internationalization of ENPs in plants and may have broad implications in foliar application of nanopesticides and minimizing contamination of food crops by ENPs.

The increased bioeconomy targets for the biomass share of renewable energy production across Europe should be met using land unsuitable for food production. Miscanthus breeding programs targeted the production of plants with a diverse range of traits allowing a wider utilization of land resources for biofuel production without competing with arable crops. These traits include increasing tolerances to drought, chilling, and to metal(loid)s excess. Two novel Miscanthus hybrids, GNT41 and GNT34, were compared against Miscanthus x giganteus (Mxg) on metal-contaminated arable land in Poland. This study aimed at evaluating their yield, biomass quality and quantifying seasonal differences in photosynthetic and transpiration parameters. A secondary objective was to identify key physiological mechanisms underlying differences in metal accumulation between the investigated plants. The new hybrids produced a similar yield to Mxg (13–15 t ha−1 yr−1), had shorter shoots, higher Leaf Area Index and stem number. Based on gas exchange measurements, GNT34 exhibited isohydric (water-conserving) behavior. The stomatal response to light of the new hybrids was at least twice as fast as that of Mxg, a trait that is often associated with increased seasonal water use efficiency. This contributed to the almost 40% reduction in shoot Pb and Cd concentrations for the new hybrids as compared to Mxg. This suggested that promoting stomatal regulation in conjunction with improved water conservation may be a target for improving plants for wider use on metals contaminated land.

Orange trees are widely cultivated in regions with high concentrations of tropospheric ozone. Citrus absorb ozone through their stomata and emit volatile organic compounds (VOC), which, together with soil emissions of NO, contribute to non-stomatal ozone removal. In a Valencia orange orchard in Exeter, California, we used fast sensors and eddy covariance to characterize water and ozone fluxes. We also measured meteorological parameters necessary to model other important sinks of ozone deposition. We present changes in magnitude of these ozone deposition sinks over the year in response to environmental parameters. Within the plant canopy, the orchard constitutes a sink for ozone, with non-stomatal ozone deposition larger than stomatal uptake. In particular, soil deposition and reactions between ozone, VOC and NO represented the major sinks of ozone. This research aims to help the development of metrics for ozone-risk assessment and advance our understanding of citrus in biosphere-atmosphere exchange.

Two common (semi-) natural temperate grassland species, Dactylis glomerata and Ranunculus acris, were grown in competition and exposed to two watering regimes: well-watered (WW, 20–40% v/v) and reduced-watered (RW, 7.5–20% v/v) in combination with eight ozone treatments ranging from pre-industrial to predicted 2100 background levels. For both species there was a significant increase in leaf damage with increasing background ozone concentration. RW had no protective effect against increasing levels of ozone-induced senescence/injury. In high ozone, based on measurements of stomatal conductance, we propose that ozone influx into the leaves was not prevented in the RW treatment, in D. glomerata because stomata were a) more widely open than those in less polluted plants and b) were less responsive to drought. Total seasonal above ground biomass was not significantly altered by increased ozone; however, ozone significantly reduced root biomass in both species to differing amounts depending on watering regime.

Plants of one evergreen oak (Quercus ilex) and three deciduous oaks (Q. faginea, with small leaves; Q. pyrenaica and Q. robur, with large leaves) were exposed both to filtered air and to enhanced ozone levels in Open-Top Chambers. Q. faginea and Q. pyrenaica were studied for the first time. Based on visible injury, gas exchange, chlorophyll content and biomass responses, Q. pyrenaica was the most sensitive species, and Q. ilex was the most tolerant, followed by Q. faginea. Functional leaf traits of the species were related to differences in sensitivity, while accumulated ozone flux via stomata (POD₁.₆) partly contributed to the observed differences. For risk assessment of Mediterranean vegetation, the diversity of responses detected in this study should be taken into account, applying appropriate critical levels.

Hydrogen (δ2H), carbon (δ13C), oxygen (δ18O) and nitrogen (δ15N) isotopes of tree rings growing in field conditions can be indicative of past pollution effects. The characteristic δ13C trend is a positive shift generally explained by invoking closure of stomata, but experimental studies suggest that increased rates of carboxylation could also generate such trends. In many cases the δ18O and δ2H values decrease in trees exposed to pollution and exhibit inverse coinciding long-term trends with δ13C values. However, some trees exposed to diffuse pollution and experimental conditions can show an increase or no δ18O change even if δ13C values increase. These diverse responses depend on how stress conditions modify physiological functions such as stomatal conductance, carboxylation, respiration, and perhaps water assimilation by the root system. Recent studies suggest that δ15N changes in trees can be caused by soil acidification and accumulation of anthropogenic emissions with isotopic signals deviating from natural N.

The impact of ozone (O3) pollution events on the plant drought response needs special attention because spring O3 episodes are often followed by summer drought. By causing stomatal sluggishness, O3 could affect the stomatal dynamic during a subsequent drought event. In this context, we studied the impact of O3 exposure and water deficit (in the presence or in the absence of O3 episode) on the stomatal closure/opening mechanisms relative to irradiance or vapour pressure deficit (VPD) variation. Two genotypes of Populus nigra x deltoides were exposed to various treatments for 21 days. Saplings were exposed to 80 ppb/day O3 for 13 days, and then to moderate drought for 7 days. The curves of the stomatal response to irradiance and VPD changes were determined after 13 days of O3 exposure, and after 21 days in the case of subsequent water deficit, and then fitted using a sigmoidal model. The main responses under O3 exposure were stomatal closure and sluggishness, but the two genotypes showed contrasting responses. During stomatal closure induced by a change in irradiance, closure was slower for both genotypes. Nonetheless, the genotypes differed in stomatal opening under light. Carpaccio stomata opened more slowly than control stomata, whereas Robusta stomata tended to open faster. These effects could be of particular interest, as stomatal impairment was still present after O3 exposure and could result from imperfect recovery. Under water deficit alone, we observed slower stomatal closure in response to VPD and irradiance, but faster stomatal opening in response to irradiance, more marked in Carpaccio. Under the combined treatment, most of the parameters showed antagonistic responses. Our results highlight that it is important to take genotype-specific responses and interactive stress cross-talk into account to improve the prediction of stomatal conductance in response to various environmental modifications.

Tree canopies play a key role in the cycling of polycyclic aromatic hydrocarbons (PAHs) in terrestrial ecosystems, as leaves can capture PAHs from the air. In this study, accumulation of PAHs was compared in an evergreen species, P. pinaster, and in a deciduous species, Q. robur, in relation to some physio-morphological characteristics. For this purpose, pine needles and oak leaves collected from different sites across Galicia (NW Spain) were analysed to determine PAH contents, specific leaf area, stomatal density and conductance.Leaves and needles contained similar total amounts of PAHs. The major contribution of particle-bound PAHs in oak (the concentrations of 4- and 5-ring PAHs were two times higher, and those of 6-ring PAHs five times higher in oak than in pine) may be related to the higher specific leaf area (13 and 4 cm2 g−1 dry mass in respectively oak and pine). However, the major contribution of vapor-phase PAHs in pines may be affected by the stomatal conductance (two times higher in pine than in oak). Moreover, an increase in the diameter at breast height of trees led to an increase in accumulation of PAHs, with pine capturing higher amounts of low and medium molecular weight PAHs. The study findings underline the potential role of trees in improving air quality, taking into account the canopy biomass and life cycle.

Stomata tend to narrow under ozone (O3) impact, leading to limitation of stomatal O3 influx. Here, we review stomatal response under recently conducted free-air O3 exposure experiments on two species of the same tree genus: Fagus sylvatica at Kranzberg Forest (Germany) and F. crenata at Sapporo Experimental Forest (Japan). Both beeches exhibited reduction in stomatal conductance (gs) by 10–20% under experimentally enhanced O3 regimes throughout the summer relative to ambient-air controls. Stomatal narrowing occurred, in early summer, in the absence of reduced carboxylation capacity of Rubisco, although photosynthetic net CO2 uptake rate temporarily reflected restriction to some minor extent. Observed stomatal narrowing was, however, diminished in autumn, suggesting gradual loss of stomatal regulation by O3. Monotonic decline in gs with cumulative O3 exposure or flux in current modeling concepts appear to be unrealistic in beech.