Ozone (O₃) is a phytotoxic air pollutant, the adverse effects of which on growth and photosynthesis are modified by other environmental factors. In this study, we examined the combined effects of O₃, elevated CO₂, and soil nitrogen supply on Siebold's beech seedlings. Seedlings were grown under combinations of two levels of O₃ (low and two times ambient O₃ concentration), two levels of CO₂ (ambient and 700 ppm), and three levels of soil nitrogen supply (0, 50, and 100 kg N ha⁻¹ year⁻¹) during two growing seasons (2019 and 2020), with leaf photosynthetic traits being determined during the second season. We found that elevated CO₂ ameliorated O₃-induced reductions in photosynthetic activity, whereas the negative effects of O₃ on photosynthetic traits were enhanced by soil nitrogen supply. We observed three-factor interactions in photosynthetic traits, with the ameliorative effects of elevated CO₂ on O₃-induced reductions in the maximum rate of carboxylation being more pronounced under high than under low soil nitrogen conditions in July. In contrast, elevated CO₂-induced amelioration of the effects of O₃ on stomatal function-related traits was more pronounced under low soil nitrogen conditions. Although we observed several two- or three-factor interactions of gas and soil treatments with respect to leaf photosynthetic traits, the shoot to root dry mass (S/R) ratio was the only parameter for which a significant interaction was detected among seedling growth parameters. O₃ caused a significant increase in S/R under ambient CO₂ conditions, whereas no similar effects were observed under elevated CO₂ conditions. Collectively, our findings reveal the complex interactive effects of elevated CO₂ and soil nitrogen supply on the detrimental effects of O₃ on leaf photosynthetic traits, and highlight the importance of taking into consideration differences between the responses of CO₂ uptake and growth to these three environmental factors.

Ozone-induced changes in the relationship between photosynthesis (Aₙ) and stomatal conductance (gₛ) vary among species, leading to inconsistent water use efficiency (WUE) responses to elevated ozone (O₃). Thus, few vegetation models can accurately simulate the effects of O₃ on WUE. Here, we conducted an experiment exposing two differently O₃-sensitive species (Cotinus coggygria and Magnolia denudata) to five O₃ concentrations and investigated the impact of O₃ exposure on predicted WUE using a coupled Aₙ-gₛ model. We found that increases in stomatal O₃ uptake caused linear reductions in the maximum rates of Rubisco carboxylation (Vcₘₐₓ) and electron transport (Jₘₐₓ) in both species. In addition, a negative linear correlation between O₃-induced changes in the minimal gₛ of the stomatal model (g₀) derived from the theory of optimal stomatal behavior and light-saturated photosynthesis was found in the O₃-sensitive M. denudata. When the O₃ dose-based responses of Vcₘₐₓ and Jₘₐₓ were included in a coupled Aₙ-gₛ model, simulated Aₙ under elevated O₃ were in good agreement with observations in both species. For M. denudata, incorporating the O₃ response of g₀ into the coupled model further improved the accuracy of the simulated gₛ and WUE. In conclusion, the modified Vcₘₐₓ, Jₘₐₓ and g₀ method presented here provides a foundation for improving the prediction for O₃-induced changes in Aₙ, gₛ and WUE.

Microbial degradation of phenol and cresols can occur under oxic and anoxic conditions by different degradation pathways. One recent technique to take insight into reaction mechanisms is compound-specific isotope analysis (CSIA). While enzymes and reaction mechanisms of several degradation pathways have been characterized in (bio)chemical studies, associated isotope fractionation patterns have been rarely reported, possibly due to constraints in current analytical methods. In this study, carbon enrichment factors and apparent kinetic isotope effects (AKIEc) of the initial steps of different aerobic and anaerobic phenol and cresols degradation pathways were analyzed by isotope ratio mass spectrometry connected with liquid chromatography (LC-IRMS). Significant isotope fractionation was detected for aerobic ring hydroxylation, anoxic side chain hydroxylation, and anoxic fumarate addition, while anoxic carboxylation reactions produced small and inconsistent fractionation. The results suggest that several microbial degradation pathways of phenol and cresols are detectable in the environment by CSIA.

Warfarin-induced vitamin K (VK) recycling impairment is used worldwide as a rodenticide and human thromboembolic prophylactic. Since VK metabolism/signaling pathways have been conserved throughout vertebrate evolution, its release to the environment might impact on aquatic organisms. Present study assessed the toxic effect of warfarin (0, 5, 25 and 125 mg L−1) on zebrafish development and characterized underlying mechanisms of action through qPCR analysis of VK-related genes. Expression of pregnane X receptor (pxr), the nuclear receptor binding vitamin K, was ubiquitous in zebrafish and suggests that warfarin exposure may interfere with several biological processes. Indeed, warfarin exposure of zebrafish larvae caused hemorrhages in brain, skeletal deformities and triggered ectopic calcifications, which may be the consequence of an altered γ-carboxylation of VK-dependent proteins and/or pxr signaling. This study provides new insights into warfarin effects as a bone homeostasis disruptor and soft tissue calcification inductor, and its potential risk for aquatic environments.

We set up a free-air ozone (O3) exposure system for determining the photosynthetic responses of Siebold's beech (Fagus crenata) and oak (Quercus mongolica var. crispula) to O3 under field conditions. Ten-year-old saplings of beech and oak were exposed to an elevated O3 concentration (60 nmol mol−1) during daytime from 6 August to 11 November 2011. Ozone significantly reduced the net photosynthetic rate in leaves of both species in October, by 46% for beech and 15% for oak. In beech there were significant decreases in maximum rate of carboxylation, maximum rate of electron transport in photosynthesis, nitrogen content and photosynthetic nitrogen use efficiency, but not in oak. Stomatal limitation of photosynthesis was unaffected by O3. We therefore concluded photosynthesis in beech is more sensitive to O3 than that in oak, and the O3-induced reduction of photosynthetic activity in beech was due not to stomatal closure, but to biochemical limitation.

The effect of elevated CO2 and O3 on apparent quantum yield (), maximum photosynthesis (Pmax), carboxylation efficiency (Vcmax) and electron transport capacity (Jmax) at different canopy locations was studied in two aspen (Populus tremuloides) clones of contrasting O3 tolerance. Local light climate at every leaf was characterized as fraction of above-canopy photosynthetic photon flux density (ÆPPFD). Elevated CO2 alone did not affect or Pmax, and increased Jmax in the O3-sensitive, but not in the O3-tolerant clone. Elevated O3 decreased leaf chlorophyll content and all photosynthetic parameters, particularly in the lower canopy, and the negative impact of O3 increased through time. Significant interaction effect, whereby the negative impact of elevated O3 was exaggerated by elevated CO2 was seen in Chl, N and Jmax, and occurred in both O3-tolerant and O3-sensitive clones. The clonal differences in the level of CO2 × O3 interaction suggest a relationship between photosynthetic acclimation and background O3 concentration. Photosynthetic acclimation to elevated CO2 depends on the background oxidant 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.

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.

The intentional production and degradation of plastic debris may result in the formation of nanoplastics. Currently, the scarce information on the environmental behaviors of nanoplastics hinders accurate assessment of their potential risks. Herein, the aggregation kinetics of different surface-modified polystyrene nanoparticles in monovalent and divalent electrolytes was investigated to shed some light on the fate of nanoplastics in the aquatic environment. Three monodisperse nanoparticles including unmodified nanoparticles (PS-Bare), carboxylated nanoparticles (PS–COOH) and amino modified nanoparticles (PS–NH₂), as well as one polydisperse nanoparticles that formed by laser ablation of polystyrene films (PS-Laser) were used as models to understand the effects of surface groups and morphology. Results showed that aggregation kinetics of negatively charged PS-Bare and PS-COOH obeyed the DLVO theory in NaCl and CaCl₂ solutions. The presence of Suwannee river natural organic matters (SRNOM) suppressed the aggregation of PS-Bare and PS-COOH in monovalent electrolytes by steric hindrance. However, in divalent electrolytes, their stability was enhanced at low concentrations of SRNOM (below 5 mg C L⁻¹), while became worse at high concentrations of SRNOM (above 5 mg C L⁻¹) due to the interparticle bridging effect caused by Ca²⁺ and carboxyl groups of SRNOM. The cation bridging effect was also observed for PS-laser in the presence of high concentrations of divalent electrolytes and SRNOM. The adsorption of SRNOM could neutralize or even reverse surface charges of positively charged PS-NH₂ at high concentrations, thus enhanced or inhibited the aggregation of PS-NH₂. No synergistic effect of Ca²⁺ and SRNOM was observed on the aggregation of PS-NH₂, probably due to the steric repulsion imparted by the surface modification. Our results highlight that surface charge and surface modification significantly influence aggregation behaviors of nanoplastics in aquatic systems.

We aimed to clarify the effects of ozone (O3) on photosynthetic ability of upper and lower canopy leaves of Fagus crenata Blume seedlings grown under different soil nutrient conditions. To accomplish this objective, we analyzed the response of photosynthetic parameters such as maximum carboxylation rate (Vcmax) to cumulative stomatal O3 uptake (ΣFst) and reduction rate of Vcmax per unit ΣFst as an index of detoxification capacity for O3. The seedlings of Fagus crenata were grown for two growing seasons (2014–2015) in nine treatments comprised of a combination of three levels of gas treatments (charcoal-filtered air or 1.0- or 1.5-times ambient O3 concentration) and three levels of soil nutrient treatments (non-fertilized or a supply of relatively low or high concentrations of compound fertilizer). The nutrient supply significantly increased the degree of O3-induced reduction in Vcmax in September. However, nutrient supply did not significantly increase ΣFst and reduce the detoxification capacity for O3. On the other hand, the degree of O3-induced reduction in Vcmax of upper canopy leaves was higher as compared with that of lower canopy leaves in August due to the higher ΣFst. However, the reduction rate of Vcmax per unit ΣFst in lower canopy leaves was higher than that in upper canopy leaves, indicating lower detoxification capacity for O3 in lower canopy leaves. Reduction rate of Vcmax per unit ΣFst over the threshold, which is assumed to be proportional to gross photosynthetic rate, was similar between upper and lower canopy leaves. Therefore, capacity of photosynthetic CO2 assimilation is likely to be associated with detoxification capacity for O3 in upper and lower canopy leaves of F. crenata seedlings grown under different soil nutrient conditions.