A flux-based assessment of the effects of ozone on foliar injury, photosynthesis, and yield of bean (Phaseolus vulgaris L. cv. Borlotto Nano Lingua di Fuoco) in open-top chambers

Stomatal ozone uptake, determined with the Jarvis' approach, was related to photosynthetic efficiency assessed by chlorophyll fluorescence and reflectance measurements in open-top chamber experiments on Phaseolus vulgaris. The effects of O₃ exposure were also evaluated in terms of visible and microscopical leaf injury and plant productivity. Results showed that microscopical leaf symptoms, assessed as cell death and H₂O₂ accumulation, preceded by 3-4 days the appearance of visible symptoms. An effective dose of ozone stomatal flux for visible leaf damages was found around 1.33 mmol O₃ m⁻². Significant linear dose-response relationships were obtained between accumulated fluxes and optical indices (PRI, NDI, ΔF/F_m^′). The negative effects on photosynthesis reduced plant productivity, affecting the number of pods and seeds, but not seed weight. These results, besides contributing to the development of a flux-based ozone risk assessment for crops in Europe, highlight the potentiality of reflectance measurements for the early detection of ozone stress.     

Lead tolerance and physiological adaptation mechanism in roots of accumulating and non-accumulating ecotypes of Sedum alfredii

Background, aim and scope

Lead (Pb) accumulation in soils affects plants primarily through their root systems. The aim of this study was to investigate early symptoms of the loss of membrane integrity and lipid peroxidation in root tissues and physiological adaptation mechanism to Pb in accumulating ecotypes (AE) and non-accumulating ecotypes (NAE) of Sedum alfredii under Pb stress in hydroponics.

Methods and results

Histochemical in situ analyses, fluorescence imaging, and normal physiological analysis were used in this study. Pb accumulation in roots of both AE and NAE increased linearly with increasing Pb levels (0?C200???M), and a significant difference between both ecotypes was noted. Both loss of plasma membrane integrity and lipid peroxidation in root tissues became serious with increasing Pb levels, maximum tolerable Pb level was 25 and 100???M for NAE and AE, respectively. Pb supplied at a toxic level caused a burst of reactive oxygen species (ROS) in root cells in both ecotypes. However, the root cells of AE had inherently higher activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), and lipoxygenase (LOX) in control plants, and the induction response of these antioxidant enzymes occurred at lower Pb level in AE than NAE. AE plants maintained higher ascorbic acid and H₂O₂ concentrations in root cells than NAE when exposed to different Pb levels, and Pb induced more increase in dehydroascorbate (DHA), catalase (CAT), and ascorbate peroxidase (APX) in AE than NAE roots.

Discussion and conclusion

Results indicate that histochemical in situ analyses of root cell death and lipid peroxidation under Pb short-term stress was sensitive, reliable, and fast. Higher tolerance in roots of accumulating ecotype under Pb stress did depend on effective free oxygen scavenging by making complex function of both constitutively higher activities and sensitive induction of key antioxidant enzymes in root cells of S. alfredii.     

Contrasting effects of ozone under different water supplies in two Mediterranean tree species

The effects of ozone (O₃) exposure under different water availabilities were studied in two Mediterranean tree species: Quercus ilex and Ceratonia siliqua. Plants were exposed to different O₃ concentrations in open top chambers (charcoal-filtered air (CF), non-filtered air (NF)) and non-filtered air plus 40 ppb_v of O₃ ((7:00–17:00 solar time) (NF+)) during 2 years, and to different water regimes (IR, sample irrigation, and WS, reduced water dose to 50%) through the last of those 2 years. AOT40 in the NF+ treatment was 59265 ppb_v h (from March 1999 to August 1999) while in the NF treatment, the AOT40 was 6727 ppb_v h for the same period. AOT40 was always 0 in the CF treatment. WS plants presented lower stomatal conductances and net photosynthetic rates, and higher foliar N concentrations than IR plants in both species. The irrigation treatment did not change the response trends to ozone in Q. ilex, the most sensitive species to O₃ ambient concentrations, but it changed those of C. siliqua, the least sensitive species, since its ozone-fumigated WS plants did not decrease their net photosynthetic rates nor their biomass accumulation as it happened to its ozone-fumigated IR plants. These results show interspecific variations in O₃ sensitivity under different water availabilities.     

Investigation of low-level <Superscript>242</Superscript>Pu contamination on nutrition disturbance and oxidative stress in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.

Plutonium associated with higher molecular weight molecules is presumed to be poorly mobile and hardly plant available. In our present study, we investigate the uptake and effects of Pu treatments on Solanum tuberosum plants in amended Hoagland medium at concentrations of [²⁴²Pu] = 100 and 500 nm, respectively. We found a direct proof of oxidative stress in the plants caused by these rather low concentrations. For the confirmation of oxidative stress, we explored the production of nitric oxide (NO) and hydrogen peroxide (H₂O₂) by epifluorescence microscopy. Oxidative stress markers like lipid peroxidation and superoxide radicals (O₂ ^??) are monitored through histochemical analysis. The biochemical parameters i.e. chlorophyll and carotenoids are measured as an indicator of cellular damage in the tested plants including the enzymatic parameters such as catalase and glutathione reductase. From our work, we conclude that Pu in low concentration has no significant effects on the uptake of many trace and macroelements. In contrast, the content of O₂ ^?? , malondialdehyde (MDA), and H₂O₂ increases with increasing Pu concentration in the solution, while the opposite effects was found for NO, catalase, and glutathione reductase. These findings prove that even low concentration of Pu regulates ROS production and generate oxidative stress in S. tuberosum L.     

Physiological and biochemical responses of <Emphasis Type="Italic">Machilus ichangensis</Emphasis> Rehd. et Wils and <Emphasis Type="Italic">Taxus chinensis</Emphasis> (Pilger) Rehd. to elevated O<Subscript>3</Subscript> in subtropical China

Considerable researches have documented the negative effects of ozone on woody species in North America and Europe; however, little is known about how woody tree species respond to elevated O₃ in subtropical China, and most of the previous studies were conducted using pot experiment. In the present study, Machilus ichangensis Rehd. et Wils (M. ichangensis) and Taxus chinensis (Pilger) Rehd. (T. chinensis), evergreen tree species in subtropical China, were exposed to non-filtered air (NF), 100 nmol mol^?1 O₃ (E1) and 150 nmol mol^?1 O₃ (E2), in open-top chambers under field conditions from 21st March to 2nd November 2015. In this study, O₃ fumigation significantly reduced net photosynthesis rate (Pn) in M. ichangensis in the three measurements and in T. chinensis in the last measurement. Also, non-stomatal factors should be primarily responsible for the decreased Pn. O₃ fumigation-induced increase in malondialdehyde, superoxide dismutase, and reduced ascorbic acid levels indicated that antioxidant defense mechanism had been stimulated to prevent O₃ stress and repair the oxidative damage. Yet, the increase of antioxidant ability was not enough to counteract the harm of O₃ fumigation. Because of the decrease in CO₂ assimilation, the growth of the two tree species was restrained ultimately. The sensitivity of the two tree species to O₃ can be determined: M. ichangensis > T. chinensis. It suggests a close link between the rising O₃ concentrations and the health risk of some tree species in subtropics in the near future.     

Lead-induced oxidative damage in steriled seedlings of Nymphoides peltatum

The effects of increasing concentrations of lead (Pb) on Pb accumulation and its influence on nutrient elements, malondialdehyde (MDA) content, generation of superoxide anion (O₂ ^?·), hydrogen peroxide (H₂O₂) content, antioxidant enzymes activities, soluble protein, and photosynthetic pigment, as well as chloroplast ultrastructure in steriled seedlings of Nymphoides peltata (S. G. Gmel.) Kuntze were investigated in order to understand Pb-induced toxicity. The accumulation of Pb was found to increase in a concentration-dependent manner. Nutrient elements (Ca, K, Fe, Mn, and Mo) were also affected. MDA content and O₂ ^?· generation rate increased progressively, while H₂O₂ content first boosted up at a low Pb concentration of 12.5 μM but then declined. Guaiacol peroxidase and catalase activities increased alternately, while superoxide dismutase activity gradually fell. Negative correlations were found between Pb and soluble protein and photosynthetic pigment. Moreover, Pb exposure resulted in a significant damage of chloroplasts. Taken together, these findings supported the hypothesis that Nymphoides peltatum underwent oxidative stress induced by Pb. In addition, both the disorder of nutrient elements and the damage to the ultrastructure of chloroplasts were indicative of general disarray in the cellular functions exerted by Pb.     

Using optical remote sensing techniques to track the development of ozone-induced stress

In this paper, a literature review about optical remote sensing (RS) of O₃ stress is presented. Studies on O₃-induced effects on vegetation reflectance have been conducted since late ‘70s based on the analysis of optical RS data. Literature review reveals that traditional RS techniques were able to detect changes in leaf and canopy reflectance related to O₃-induced stress when visible symptoms already occurred. Only recently, advanced RS techniques using hyperspectral sensors, demonstrated the feasibility of detecting the stress in its early phase by monitoring excess energy dissipation pathways such as chlorophyll fluorescence and non-photochemical quenching (NPQ). Steady-state fluorescence (Fs), measured by exploiting the Fraunhofer line depth principle and NPQ related xanthophyll-cycle, estimated through the photochemical reflectance index (PRI) responded to O₃ fumigation before visible symptoms occurred. This opens up new possibilities for the early detection of vegetation O₃ stress by means of hyperspectral RS.     

Volatile organic compounds from Italian vegetation and their interaction with ozone

Volatile Organic Compounds (VOCs) emitted from vegetation (particularly isoprenoids) represent an important source of atmospheric hydrocarbons almost double the anthropogenic source. When biogenic VOC mix with NO_x in the presence of UV radiation, ozone (O₃) is formed. In Italy, optimal conditions for O₃ formation in terms of VOC/NO_x ratios and abundance of UV radiation occur for long periods of the year. Moreover, Italian vegetation includes several species that are strong and evergreen isoprenoid emitters, and high temperatures for part of the year further stimulate these temperature-dependent emissions. We review emission of isoprenoids from Italian vegetation, current knowledge on the impact of rising O₃ levels on isoprenoid emission, and evidence showing that isoprenoids can increase both the O₃ flux to the plant and protection against oxidative stress because of their antioxidant functions. This trait not only influences plant tolerance to O₃ but also may substantially alter the flux of O₃ between atmosphere and biosphere.     

Impact of nano-CuO stress on rice (Oryza sativa L.) seedlings

Indiscriminate release of metal oxide nanoparticles (NPs) into the environment due to anthropogenic activities has become a serious threat to the ecological system including plants. The present study assesses the toxicity of nano-CuO on rice (Oryza sativa cv. Swarna) seedlings. Three different levels of stress (0.5 mM, 1.0 mM and 1.5 mM suspensions of copper II oxide, <50 nm particle size) were imposed and seedling growth performance was studied along control at 7 and 14 d of experiment. Modulation of ascorbate–glutathione cycle, membrane damage, in vivo ROS detection, foliar H₂O₂ and proline accumulation under nano-CuO stress were investigated in detail to get an overview of nano-stress response of rice. Seed germination percentage was significantly reduced under stress. Higher uptake of Evans blue by nano-CuO stressed roots over control indicates loss of root cells viability. Presence of dark blue and deep brown spots on leaves evident after histochemical staining with NBT and DAB respectively indicate severe oxidative burst under nano-copper stress. APX activity was found to be significantly increased in 1.0 and 1.5 mM CuO treatments. Nevertheless, elevated APX activity might be insufficient to scavenge all H₂O₂ produced in excess under nano-CuO stress. That may be the reason why stressed leaves accumulated significantly higher H₂O₂ instead of having enhanced APX activity. In addition, increased GR activity coupled with isolated increase in GSH/GSSG ratio does not seem to prevent cells from oxidative damages, as evident from higher MDA level in leaves of nano-CuO stressed seedlings over control. Enhanced proline accumulation also does not give much protection against nano-CuO stress. Decline in carotenoids level might be another determining factor of meager performance of rice seedlings in combating nano-CuO stress induced oxidative damages.     

Biological effects of palytoxin-like compounds from Ostreopsis cf. ovata: A multibiomarkers approach with mussels Mytilus galloprovincialis

Massive blooms of the harmful benthic dinoflagellate Ostreopsis cf. ovata are of growing environmental concern in the Mediterranean, having recently caused adverse effects on benthic invertebrates and also some intoxication episodes to humans.The toxicological potential of produced palytoxin-like compounds was investigated in the present study on a typical marine sentinel species, the mussel Mytilus galloprovincialis. Organisms were sampled during various phases of a O. cf. ovata bloom, in two differently impacted sites. The presence of the algal toxins was indirectly assessed in mussels tissues (mouse test and hemolysis neutralization assay), while biological and toxicological effects were evaluated through the measurement of osmoregulatory and neurotoxic alterations (Na⁺/K⁺-ATPase and acetylcholinesterase activities), oxidative stress responses (antioxidant defences and total oxyradical scavenging capacity), lipid peroxidation processes (level of malondialdehyde), peroxisomal proliferation, organelle dysfunctions (lysosomal membrane stability, accumulation of lipofuscin and neutral lipids), immunological impairment (granulocytes percentage).Obtained results demonstrated a significant accumulation of algal toxins in mussels exposed to O. cf. ovata. These organisms exhibited a marked inhibition of the Na⁺/K⁺-ATPase activity and alterations of immunological, lysosomal and neurotoxic responses. Markers of oxidative stress showed more limited variations suggesting that toxicity of the O. cf. ovata toxins is not primarily mediated by an over production of reactive oxygen species. This study provided preliminary results on the usefulness of a multi-biomarker approach to assess biological alterations and toxicological events associated to blooms of O. cf. ovata in marine organisms.     

The efficiency of tobacco Bel-W3 and native species for ozone biomonitoring in subtropical climate, as revealed by histo-cytochemical techniques

We aimed to verify whether hydrogen peroxide (H₂O₂) accumulation and cell death are detected early in three bioindicators of ozone (O₃), Nicotiana tabacum ‘Bel-W3’, Ipomoea nil ‘Scarlet O’Hara’ and Psidium guajava ‘Paluma’, and whether environmental factors also affect those microscopic markers. The three species were exposed to chronic levels of O₃ in a subtropical area and a histo-cytochemical technique that combines 3,3′-diaminobenzidine (DAB) with Evans blue staining was used in the assessments. The three species accumulated H₂O₂, but a positive correlation with O₃ concentration was only observed in N. tabacum. A positive correlation between O₃ and cellular death was also observed in N. tabacum. In I. nil and P. guajava, environmental factors were responsible for symptoms at the microscopic level, especially in P. guajava. We conclude that the most appropriate and least appropriate bioindicator plant for O₃ monitoring in the subtropics are N. tabacum ‘Bel-W3’ and P. guajava ‘Paluma’, respectively.     

Physiological and biochemical mechanisms of spermine-induced cadmium stress tolerance in mung bean (<Emphasis Type="Italic">Vigna radiata</Emphasis> L.) seedlings

The role of exogenous spermine (0.25 mM Spm, a type of polyamine (PA) in reducing Cd uptake and alleviating Cd toxicity (containing 1 and 1.5 mM CdCl₂ in the growing media) effects was studied in the mung bean (Vigna radiata L. cv. BARI Mung-2) plant. Exogenously applied Spm reduced Cd content, accumulation, and translocation in different plant parts. Increasing phytochelatin content, exogenous Spm reduced Cd accumulation and translocation. Spm application reduced the Cd-induced oxidative damage which was reflected from the reduction of H₂O₂ content, O₂ ^?– generation rate, lipoxygenase (LOX) activity, and lipid peroxidation level and also reflected from the reduction of spots of H₂O₂ and O₂ ^?– from mung bean leaves (compared to control treatment). Spm pretreatment increased non-enzymatic antioxidant contents (ascorbate, AsA, and glutathione, GSH) and activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) which reduced oxidative stress. The cytotoxicity of methylglyoxal (MG) is also reduced by exogenous Spm because it enhanced glyoxalase system enzymes and components. Through osmoregulation, Spm maintained a better water status of Cd-affected mung bean seedlings. Spm prevented the chl damage and increased its content. Exogenous Spm also modulated the endogenous free PAs level which might have the roles in improving physiological processes including antioxidant capacity, osmoregulation, and Cd and MG detoxification capacity. The overall Spm-induced tolerance of mung bean seedlings to Cd toxicity was reflected through improved growth of mung bean seedlings.     

Exogenously applied calcium alleviates cadmium toxicity in Matricaria chamomilla L. plants

Cadmium (Cd) toxicity in plants leads to serious disturbances of physiological processes, such as inhibition of chlorophyll synthesis, oxidative injury to the plant cells and water and nutrient uptake. Response of Matricaria chamomilla L. to calcium chloride (CaCl₂) enrichment in growth medium for reducing Cd toxicity were studied in this study. Hydroponically cultured seedlings were treated with 0, 0.1, 1, and 5 mM CaCl₂, under 0, 120, and 180 μM CdCl₂ conditions, respectively. The study included measurements pertaining to physiological attributes such as growth parameters, Cd concentration and translocation, oxidative stress, and accumulation of phenolics. Addition of CaCl₂ to growth media decreased the Cd concentration, activity of antioxidant enzymes, and reactive oxygen species accumulation in the plants treated with different CdCl₂, but increased the growth parameters. Malondialdehyde and total phenolics in shoots and roots were not much affected when plants were treated only with different CaCl₂ levels, but it showed a rapid increase when the plants were exposed to 120 and 180 CdCl₂ levels. CaCl₂ amendment also ameliorated the CdCl₂-induced stress by reducing oxidative injury. The beneficial effects of CaCl₂ in ameliorating CdCl₂ toxicity can be attributed to the Ca-induced reduction of Cd concentration, by reducing the cell-surface negativity and competing for Cd²⁺ ion influx, activity enhancement of antioxidant enzymes, and biomass accumulation.     

Oxidative stress status,antioxidant metabolism and polypeptide patterns in Juncus maritimus shoots exhibiting differential mercury burdens in Ria de Aveiro coastal lagoon (Portugal)

This study assessed the oxidative stress status, antioxidant metabolism and polypeptide patterns in salt marsh macrophyte Juncus maritimus shoots exhibiting differential mercury burdens in Ria de Aveiro coastal lagoon at reference and the sites with highest, moderate and the lowest mercury contamination. In order to achieve these goals, shoot-mercury burden and the responses of representative oxidative stress indices, and the components of both non-glutathione- and glutathione-based H₂O₂-metabolizing systems were analyzed and cross-talked with shoot-polypeptide patterns. Compared to the reference site, significant elevations in J. maritimus shoot mercury and the oxidative stress indices such as H₂O₂, lipid peroxidation, electrolyte leakage and reactive carbonyls were maximum at the site with highest followed by moderate and the lowest mercury contamination. Significantly elevated activity of non-glutathione-based H₂O₂-metabolizing enzymes such as ascorbate peroxidase and catalase accompanied the studied damage-endpoint responses, whereas the activity of glutathione-based H₂O₂-scavenging enzymes glutathione peroxidase and glutathione sulfo-transferase was inhibited. Concomitantly, significantly enhanced glutathione reductase activity and the contents of both reduced and oxidized glutathione were perceptible in high mercury-exhibiting shoots. It is inferred that high mercury-accrued elevations in oxidative stress indices were obvious, where non-glutathione-based H₂O₂-decomposing enzyme system was dominant over the glutathione-based H₂O₂-scavenging enzyme system. In particular, the glutathione-based H₂O₂-scavenging system failed to coordinate with elevated glutathione reductase which in turn resulted into increased pool of oxidized glutathione and the ratio of oxidized glutathione-to-reduced glutathione. The substantiation of the studied oxidative stress indices and antioxidant metabolism with approximately 53-kDa polypeptide warrants further studies.     

Ozone sensitivity of the Mediterranean terophyte Trifolium striatum is modulated by soil nitrogen content

The influence of nitrogen (N) fertilization on the sensitivity to ozone (O₃) has been studied using the O₃ sensitive species Trifolium striatum, an annual species frequently found in therophitic grasslands of the dehesas in the central Iberian Peninsula. The experiment was carried out in open-top chambers using three different O₃ exposure treatments: charcoal-filtered air (CFA), non-filtered air (NFA) and non-filtered air plus 40 nl l⁻¹ added over ambient concentrations (NFA+). After 30 days of exposure to the different treatments, plants were placed under ambient conditions until seed ripeness was completed. Three N fertilization treatments were established receiving final doses of 10 (low), 30 (medium) or 60 (high) kg N ha⁻¹ throughout the growing period divided into four proportional doses at every 15 days. Ozone affected phenology and reduced total aerial and flower biomass and seed production in NFA and NFA+ treatments. Carry-over effects of O₃ on the reproductive ability of this species were found resulting in reductions in flower biomass and seed production detected 1 month after O₃ exposure. By contrast, N fertilization increased aerial and flower biomass. Significant O₃×N interactive effects were detected since N fertilization counterbalanced O₃-induced effects only when plants were exposed to moderate O₃ levels (NFA) but not under high O₃ concentrations (NFA+). The observed O₃ effects on reproductive ability of T. striatum might determine changes in species viability and future diversity of the ecosystem.     

Intraspecific responses of six Indian clover cultivars under ambient and elevated levels of ozone

Intraspecific variation in six cultivars of clover Trifolium alexandrinum L., (Bundel, Wardan, JHB-146, Saidi, Fahli, and Mescavi) has been studied with ambient and elevated O₃ (ambient?+?10 ppb O₃) in open top chambers. Significant effect of elevated O₃ was detected on different morphological, physiological, and biochemical parameters depicting differential response among the test cultivars. Results showed that the magnitude of O₃ induced foliar injury symptoms varied in all the cultivars. Ozone significantly depressed photosynthetic rate, stomatal conductance, and photosynthetic efficiency, although variations were cultivar specific. Ozone treatment diminished total biomass of all the cultivars; reduction was highest in Wardan with least O₃ resistance followed by Bundel, JHB-146, Saidi, Mescavi, and Fahli. According to the cumulative sensitive index, variations in the sensitivity showed that two cultivars (Wardan and Bundel) were sensitive to elevated O₃, while other three cultivars (Fahli, Saidi, and Mescavi) were resistant, and JHB-146 showed intermediate sensitivity. Therefore, the present study supported the selection of sensitive cultivar of clover as a bioindicator for O₃ under Indian conditions for the areas experiencing higher concentrations of O₃.     

Enhanced nitrogen deposition exacerbates the negative effect of increasing background ozone in Dactylis glomerata, but not Ranunculus acris

The combined impacts of simulated increased nitrogen (N) deposition (75 kg N ha⁻¹ yr⁻¹) and increasing background ozone (O₃) were studied using two mesotrophic grassland species (Dactylis glomerata and Ranunculus acris) in solardomes, by means of eight O₃ treatments ranging from 15.5 ppb to 92.7 ppb (24 h average mean). A-C_i curves were constructed for each species to gauge effects on photosynthetic efficiency and capacity, and effects on biomass partitioning were determined after 14 weeks. Increasing the background concentration of O₃ reduced the healthy above ground and root biomass of both species, and increased senesced biomass. N fertilisation increased biomass production in D. glomerata, and a significantly greater than additive effect of O₃ and N on root biomass was evident. In contrast, R. acris biomass was not affected by high N. The study shows the combined effects of these pollutants have differential implications for carbon allocation patterns in common grassland species.     

Gene expression responses of paper birch (Betula papyrifera) to elevated CO2 and O3 during leaf maturation and senescence

Gene expression responses of paper birch (Betula papyrifera) leaves to elevated concentrations of CO₂ and O₃ were studied with microarray analyses from three time points during the summer of 2004 at Aspen FACE. Microarray data were analyzed with clustering techniques, self-organizing maps, K-means clustering and Sammon's mappings, to detect similar gene expression patterns within sampling times and treatments. Most of the alterations in gene expression were caused by O₃, alone or in combination with CO₂. O₃ induced defensive reactions to oxidative stress and earlier leaf senescence, seen as decreased expression of photosynthesis- and carbon fixation-related genes, and increased expression of senescence-associated genes. The effects of elevated CO₂ reflected surplus of carbon that was directed to synthesis of secondary compounds. The combined CO₂ + O₃ treatment resulted in differential gene expression than with individual gas treatments or in changes similar to O₃ treatment, indicating that CO₂ cannot totally alleviate the harmful effects of O₃.     

DNA damage in Populus tremuloides clones exposed to elevated O3

The effects of elevated concentrations of atmospheric tropospheric ozone (O₃) on DNA damage in five trembling aspen (Populus tremuloides Michx.) clones growing in a free-air enrichment experiment in the presence and absence of elevated concentrations of carbon dioxide (CO₂) were examined. Growing season mean hourly O₃ concentrations were 36.3 and 47.3 ppb for ambient and elevated O₃ plots, respectively. The 4th highest daily maximum 8-h ambient and elevated O₃ concentrations were 79 and 89 ppb, respectively. Elevated CO₂ averaged 524 ppm (+150 ppm) over the growing season. Exposure to O₃ and CO₂ in combination with O₃ increased DNA damage levels above background as measured by the comet assay. Ozone-tolerant clones 271 and 8L showed the highest levels of DNA damage under elevated O₃ compared with ambient air; whereas less tolerant clone 216 and sensitive clones 42E and 259 had comparably lower levels of DNA damage with no significant differences between elevated O₃ and ambient air. Clone 8L was demonstrated to have the highest level of excision DNA repair. In addition, clone 271 had the highest level of oxidative damage as measured by lipid peroxidation. The results suggest that variation in cellular responses to DNA damage between aspen clones may contribute to O₃ tolerance or sensitivity.     

Copper bioaccumulation,photosystem II functioning,and oxidative stress in the seagrass <Emphasis Type="Italic">Cymodocea nodosa</Emphasis> exposed to copper oxide nanoparticles

Photosynthetic activity, oxidative stress, and Cu bioaccumulation in the seagrass Cymodocea nodosa were assessed 4, 12, 24, 48, and 72 h after exposure to two copper oxide nanoparticle (CuO NP) concentrations (5 and 10 mg L^?1). CuO NPs were characterized by scanning electron microscopy (SEM) and dynamic light scattering measurements (DLS). Chlorophyll fluorescence analysis was applied to detect photosystem II (PSII) functionality, while the Cu accumulation kinetics into the leaf blades was fitted to the Michaelis-Menten equation. The uptake kinetics was rapid during the first 4 h of exposure and reached an equilibrium state after 10 h exposure to 10 mg L^?1 and after 27 h to 5 mg L^?1 CuO NPs. As a result, 4-h treatment with 5 mg L^?1 CuO NPs, decreased the quantum yield of PS II photochemistry (Φ _PSΙΙ ) with a parallel increase in the regulated non-photochemical energy loss in PSII (Φ _NPQ ). However, the photoprotective dissipation of excess absorbed light energy as heat, through the process of non-photochemical quenching (NPQ), did not maintain the same fraction of open reaction centers (q _p ) as in control plants. This reduced number of open reaction centers resulted in a significant increase of H₂O₂ production in the leaf veins serving possibly as an antioxidant defense signal. Twenty-four-hour treatment had no significant effect on Φ _PSΙΙ and q _p compared to controls. However, 24 h exposure to 5 mg L^?1 CuO NPs increased the quantum yield of non-regulated energy loss in PSII (Φ _NO ), and thus the formation of singlet oxygen (¹O₂) via the triplet state of chlorophyll, possible because the uptake kinetics had not yet reached the equilibrium state as did 10 mg L^?1. Longer-duration treatment (48 and 72 h) had less effect on the allocation of absorbed light energy at PSII and the fraction of open reaction centers, compared to 4-h treatment, suggesting the function of a stress defense mechanism. The response of C. nodosa leaves to CuO NPs fits the “Threshold for Tolerance Model” with a threshold time (more than 4 h) required for induction of a stress defense mechanism, through H₂O₂ production.