PFOA accumulation in the leaves of basil (Ocimum basilicum L.) and its effects on plant growth, oxidative status, and photosynthetic performance.

BACKGROUND: Perfluoroalkyl substances (PFASs) are emerging contaminants of increasing concern due to their presence in the environment, with potential impacts on ecosystems and human health. These substances are considered "forever chemicals" due to their recalcitrance to degradation, and their accumulation in living organisms can lead to varying levels of toxicity based on the compound and species analysed. Furthermore, concerns have been raised about the possible transfer of PFASs to humans through the consumption of edible parts of food plants. In this regard, to evaluate the potential toxic effects and the accumulation of perfluorooctanoic acid (PFOA) in edible plants, a pot experiment in greenhouse using three-week-old basil (Ocimum basilicum L.) plants was performed adding PFOA to growth substrate to reach 0.1, 1, and 10 mg Kg- 1 dw. RESULTS: After three weeks of cultivation, plants grown in PFOA-added substrate accumulated PFOA at different levels, but did not display significant differences from the control group in terms of biomass production, lipid peroxidation levels (TBARS), content of α-tocopherol and activity of ascorbate peroxidase (APX), catalase (CAT) and guaiacol peroxidase (POX) in the leaves. A reduction of total phenolic content (TPC) was instead observed in relation to the increase of PFOA content in the substrate. Furthermore, chlorophyll content and photochemical reflectance index (PRI) did not change in plants exposed to PFAS in comparison to control ones. Chlorophyll fluorescence analysis revealed an initial, rapid photoprotective mechanism triggered by PFOA exposure, with no impact on other parameters (Fv/Fm, ΦPSII and qP). Higher activity of glutathione S-transferase (GST) in plants treated with 1 and 10 mg Kg- 1 PFOA dw (30 and 50% to control, respectively) paralleled the accumulation of PFOA in the leaves of plants exposed to different PFOA concentration in the substrate (51.8 and 413.9 ng g- 1 dw, respectively). CONCLUSION: Despite of the absorption and accumulation of discrete amount of PFOA in the basil plants, the analysed parameters at biometric, physiological and biochemical level in the leaves did not reveal any damage effect, possibly due to the activation of a detoxification pathway likely involving GST.

Assessment of physiological and biochemical responses, metal tolerance and accumulation in two eucalypt hybrid clones for phytoremediation of cadmium-contaminated waters.

Eucalyptus is a promising species for ecological restoration but plant performances under environmental constraints need to be better investigated. In particular, the toxic effects of metals on this plant species are poorly described in the literature. In this work, morpho-physiological and biochemical responses to cadmium were analysed in two eucalypt genotypes (hybrid clones of Eucalyptus camaldulensis × Eucalyptus globulus ssp. bicostata J.B. Kirkp named Velino ex 7 and Viglio ex 358) exposed for 3 weeks to 50 µM CdSO4 under hydroponics. The two eucalypt clones showed a different sensitivity to the metal. The growth reduction caused by cadmium was less than 30% in clone Velino and about 50% in clone Viglio. Cadmium mostly accumulated in plant roots and, to a lesser extent, in stem, as highlighted by the low translocation factor (Tf) measured in both clones. Net photosynthesis measurement, chlorophyll fluorescence images, transpiration values and chlorophyll content revealed a cadmium-induced impairment of physiological processes at the leaf level, which was more evident in clone Viglio. Metal binding and antioxidative compound content was differentially affected by cadmium exposure in the two eucalypt clones. Particularly, the content of thiols like cysteine and glutathione, organic acids like oxalate and citrate, and polyamines were markedly modulated in plant organs by metal treatment and highlighted different defence responses between the clones. Cadmium tolerance and accumulation ability of the eucalypt clones were evaluated and the potential of E. camaldulensis for the reclamation of metal polluted-waters is discussed.

Morpho-physiological and molecular responses of Lepidium sativum L. seeds induced by bismuth exposure.

Bismuth (Bi) is considered a "green metal" as its toxicity has been reported to be lower than other metals, particularly lead. Even though the low presence in the environment, an increase of Bi concentrations in soil and wastewater is predictable due to its enhanced uses for many industrial and medical applications. Therefore, given the little literature on the matter, particularly in plants, information on the effects of Bi on living organisms is needed. In this study, seeds of garden cress (Lepidium sativum L.), a model plant for ecotoxicological assays (OECD), were exposed to increasing Bi concentrations (0 to 485 mg L-1 Bi(NO3)3·5H2O in deionised water) in petri plates. After 72 h, the percent germination index (GI%) revealed no effects at the lowest Bi concentrations, while a slight toxicity occurred at 242 and 485 mg L-1 Bi nitrate. A significant reduction of the root length was observed in Bi-treated seedlings, especially at the highest Bi concentrations. Consistently, the Alkaline Comet Assay revealed a genotoxic effect induced by Bi exposure in garden cress seedlings. A Bi concentration-dependent metal accumulation in plantlets was also observed, with a Bi concentration higher than 1200 mg kg-1 found in plantlets at the highest Bi concentration assayed. The toxicity effects observed in the study were discussed, as contribution to the expansion of knowledge on Bi ecotoxicity and genotoxicity in plants.

Ecotoxicological and genotoxic effects of dimethyl phthalate (DMP) on Lemna minor L. and Spirodela polyrhiza (L.) Schleid. plants under a short-term laboratory assay.

The environmental occurrence of phthalates (PAE) is of great concern for the ecosystem and human health. Despite of their recognized toxicity on biota, a lack of knowledge is still present about the effects of PAE on plants. In this scenario, the effects of dimethyl phthalate (DMP) on duckweed plants (Lemna minor L. and Spirodela polyrhiza (L.) Schleid.), two model plant species for ecotoxicological and trophic studies, were investigated. Under a 7-day lab assay, morphological (biometric indicators), physiological (pigment content and photosynthetic performance) and molecular (DNA damage) parameters were studied. No effects were observed at growth and physiological level in both plants at 3 and 30 mg/L DMP. On the contrary, at 600 mg/L DMP, a concentration used for plant acute toxicity studies, a remarkable growth inhibition and pigment content and photosynthetic parameters reduction compared to control were observed in both plants species, particularly in Spirodela. Alkaline Comet assay in 24 h-treated plants revealed a genotoxic damage induced by DMP, particularly relevant in Spirodela. These results described for the first time the adverse effects exerted by DMP on aquatic plants, contributing to highlight the environmental risk associated to the presence of this compound in the aquatic ecosystem.

Catechol-Loading Nanofibrous Membranes for Eco-Friendly Iron Nutrition of Plants.

Modern agriculture requires more efficient and low-impact products and formulations than traditional agrochemicals to improve crop yields. Iron is a micronutrient essential for plant growth and photosynthesis, but it is mostly present in insoluble forms in ecosystems so that it is often limiting for plants. This study was aimed at combining natural strategies and biodegradable nanostructured materials to create environmentally friendly and low-toxic bioactive products capable of both supplying iron to Fe-deficient plants and reducing the impact of agricultural products on the environment. Consequently, free-standing electrospun nanofibrous polycaprolactone/polyhydroxybutyrate thin membranes loaded with catechol (CL-NMs) as an iron-chelating natural agent (at two concentrations) were fabricated on purpose to mobilize Fe from insoluble forms and transfer it to duckweed (Lemna minor L.) plants. The effectiveness of CL-NMs in providing iron to Fe-deficient plants, upon catechol release, tested in duckweeds grown for 4 days under controlled hydroponic conditions, displayed temporal variations in both photosynthetic efficiency and biometric parameters measured by chlorophyll fluorescence and growth imaging. Duckweeds supplied with CL-NMs hosting higher catechol concentrations recovered most of the physiological and growth performances previously impaired by Fe limitation. The absence of short-term toxicity of these materials on duckweeds also proved the low impact on ecosystems of these products.

Evaluation of morpho-physiological traits and contaminant accumulation ability in Lemna minor L. treated with increasing perfluorooctanoic acid (PFOA) concentrations under laboratory conditions.

There is increasing concern about the effects of releasing emerging contaminants (i.e. endocrine-disrupting chemicals, pharmaceuticals, personal-care products and flame retardants) into the environment. Particular attention is being paid to perfluoroalkyl substances (PFAS) because of their persistence and bioaccumulation, especially in the aquatic environment. In this paper, we present results of a study aimed at evaluating the effects of different perfluorooctanoic acid (PFOA) concentrations (2, 20 and 200 µg/L) on morpho-physiological traits in Lemna minor L. plants. The accumulation of PFOA in the plant's tissues was also monitored. L. minor was selected as a model plant for ecotoxicological studies, and we performed a seven-day assay for this investigation. The results highlight the lack of inhibitory effects on biometric parameters such as mean frond area, total frond number, multiplication rate, doubling time of frond number and average specific growth rate, for each of tested PFOA concentrations. Also, at photosynthetic level, physiological measurements showed that chlorophyll content and electron transport rate (ETR) were not affected by the exposure to PFOA. Remarkably, the chlorophyll fluorescence images, used for the first time in a study on PFOA, evidenced no impairment to the photosynthetic efficiency, measured by the maximum quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), the quantum efficiency of PSII photochemistry (ΦPSII) and the non-photochemical quenching (NPQ) over the leaf surface of PFOA-treated plants, in comparison to control. Quantification of PFOA in the growth medium at the end of the seven-day test revealed no statistically different concentrations in plates with or without L. minor plants. We detected increasing PFOA accumulation in plant tissues, in accordance with the PFOA concentrations in the medium. Therefore, the L. minor plants were capable of taking up and accumulating PFOA. The ecological impact of the environmentally relevant PFOA concentrations tested in this work on biological organisms of the aquatic environment is discussed.

Transcription Factor Networks in Leaves of Cichorium endivia: New Insights into the Relationship Between Photosynthesis and Leaf Development.

Cichorium endivia is a leafy crop closely related to Lactuca sativa that comprises two major botanical varieties characterized by a high degree of intraspecific morphological variation: var. latifolium with broad leaves (escarole) and var. crispum with narrow crisp curly leaves (endive). To investigate the relationship between leaf morphology and photosynthetic activity, escaroles and endives were used as a crop model due to the striking morphological diversity of their leaves. We constructed a leaf database for transcription factors (TFs) and photosynthesis-related genes from a refined C. endivia transcriptome and used RNA-seq transcriptomic data from leaves of four commercial endive and escarole cultivars to explore transcription factor regulatory networks. Cluster and gene co-expression network (GCN) analyses identified two main anticorrelated modules that control photosynthesis. Analysis of the GCN network topological properties identified known and novel hub genes controlling photosynthesis, and candidate developmental genes at the boundaries between shape and function. Differential expression analysis between broad and curly leaves suggested three novel TFs putatively involved in leaf shape diversity. Physiological analysis of the photosynthesis properties and gene expression studies on broad and curly leaves provided new insights into the relationship between leaf shape and function.

Effects of a ladle furnace slag added to soil on morpho-physiological and biochemical parameters of Amaranthus paniculatus L. plants.

Industrial slag from steelwork activities is considered a by-product by the EU legislation and it can be used for civil construction. In this work, an experiment in a greenhouse was conducted over a 6-week period to investigate the effect of soil enrichment with ladle furnace slag on morpho-physiological parameters of Amaranthus paniculatus L. plants. Results showed that the addition of 5% (w/w) slag to soil did not alter the plant growth, highlighting a high tolerance to this slag concentration. Contrarily, plants cultivated in a soil with 10% (w/w) slag showed a marked reduction both in growth and biometric parameters. Moreover, plants grown on a slag-rich soil (20% w/w) highlighted a very low survival rate. This behaviour was confirmed by the biochemical and physiological investigations on chlorophyll a and b content, gas exchange and chlorophyll fluorescence analyses. Metal(loid)s determination showed the accumulation of Ni, Se, Sn, As, Sb and Cd in 10% slag-treated plants, while revealed an increase in Ni, Cd, As and Pb in 5% slag-treated plants. Results are discussed highlighting the profitability of the cultivation of Amaranthus plants on slag enriched soil, as this plant species is largely used both as feedstock for energy production and for environmental restoration.

Investigation on metal tolerance and phytoremoval activity in the poplar hybrid clone "Monviso" under Cu-spiked water: Potential use for wastewater treatment.

A serious concern for the environmental and human health is represented by the increasing copper (Cu) occurrence in agricultural soils and waters, because of the possible food contamination and bioaugmentation along the trophic chain. The request for the decontamination of different matrices with an environmentally sustainable technology as the phytoremediation should be addressed by selecting plant materials with improved pollutant tolerance and removal capability. With this purpose, plants of the hybrid poplar clone "Monviso" (Populus×generosa A. Henry×P. nigra L.) were grown in growth chamber under hydroponics and exposed to excess Cu concentrations (T1, 75µM Cu; T2, 150µM Cu), selected as about 5 and 10 times higher than those allowed by the Italian regulation on water use. Results evidenced a notable Cu tolerance by poplar plants, particularly at the lowest Cu concentration. At organ level, the root system was the most affected by Cu treatment, especially in T2-exposed plants. Copper determinations revealed that the metal was mostly bioaccumulated in the roots, with a limited amount reaching the shoots. Chlorophyll content and fluorescence analyses confirmed the visible symptoms in leaves, highlighting a good physiological status in T1-exposed plants. Contrarily, an impairment of the main processes associated to photosynthesis was observed in T2-exposed plants also by gas exchange measurements. Remarkably, the Cu content analysis of the spiked water solutions revealed that poplar plants succeeded in removing almost the 50% of the total Cu amount added. These results strengthen the evidence that poplar plants represent a useful eco-friendly bio-tool for the decontamination of metal polluted waters.

Evaluation of nickel tolerance in Amaranthus paniculatus L. plants by measuring photosynthesis, oxidative status, antioxidative response and metal-binding molecule content.

Among metals, Ni has been indicated as one of the most dangerous for the environment, and plants exposed to this metal are frequently reported to undergo a severe stress condition. In this work, the tolerance responses to different Ni concentrations at physiological and biochemical levels were evaluated in Amaranthus paniculatus L., a plant species previously characterised for their ability to phytoremove Ni from metal-spiked water. Results indicated a good metal tolerance of this plant species at environmentally relevant Ni concentrations, while clear symptoms of oxidative damages were detected at higher Ni concentrations, both in roots and leaves, by measuring lipid peroxide content. At the photosynthetic level, pigment content determination, chlorophyll fluorescence image analysis and gas-exchange parameter measurements revealed a progressive impairment of the photosynthetic machinery at increasing Ni concentrations in the solution. Regarding biochemical mechanisms involved in antioxidative defence and metal binding, antioxidative enzyme (ascorbate peroxidase, APX; catalase, CAT; guaiacol peroxidase, GPX; superoxide dismutase, SOD) activity, polyamine (PA) content, polyamine oxidase (PAO) activity and organic acid (OA) content were differently affected by Ni concentration in the growth solution. A role for GPX, SOD, PAs, and oxalic and citric acid in Ni detoxification is suggested. These results can contribute to elucidate the tolerance mechanisms carried out by plants when facing environmentally relevant Ni concentrations and to identify some traits characterising the physiological and biochemical responses of Amaranthus plants to the presence and bioaccumulation of Ni.

Growth, physiological response and phytoremoval capability of two willow clones exposed to ibuprofen under hydroponic culture.

Ibuprofen (IBU) is one of the most widespread pharmaceuticals in the aquatic ecosystem, despite the high removal rate that occurs in wastewater treatment plants. Phytoremediation represents a technology to improve the performance of existing wastewater treatment. This study was conducted under hydroponics to evaluate the ability of Salicaceae plants to tolerate and reduce IBU concentration in contaminated water. To this end, we combined growth, physiological and biochemical data to study the effects of different IBU concentrations on two clones of Salix alba L. Data demonstrated that clone SS5 was more tolerant and showed a higher ability to reduce IBU concentration in the solution than clone SP3. The high tolerance to IBU shown by SS5 was likely due to several mechanisms including the capacity to maintain an elevated photosynthetic activity and an efficient antioxidative defence. These results illustrate the remarkable potential of willow to phytoremediate IBU-contaminated waters in natural and constructed wetlands.

Assessment of ibuprofen tolerance and removal capability in Populus nigra L. by in vitro culture.

Release of pharmaceuticals in the environment has been emerging as a great concern for ecosystem and human health. Ibuprofen (IBU) represents one of the most widespread pharmaceuticals in surface waters and sediments in spite of the high removal rates occurring in conventional wastewater technologies. To assess the potentiality of phytoremediation in assisting these technologies, the screening of plant species for tolerance and removal ability of pollutants is a very important issue. In this study, the effects of different IBU concentrations on callus cultures of Populus nigra L., a pioneer tree species in the riparian ecosystem, were investigated. Results evidenced a notable tolerance of poplar cells to IBU, especially at high concentrations (IBU 30 mg L(-1)), which even stimulated growth. At this concentration, the ability to withstand IBU was accompanied by inhibition of lipoxygenase (LOX) activity, reduction of lipid peroxide content and increase of membrane redox activity. Irrespective of initial IBU concentrations, a complete removal of this compound from the growth medium by poplar cells during a subculture occurred. Antioxidative enzyme activities and polyamine content were stimulated by IBU 0.03 mg L(-1), while no effect was found in cells exposed to IBU 30 mg L(-1), except for a decrease of guaiacol peroxidase (GPX) activity. These findings put in evidence a notable potential of this plant species for the phytoremediation of IBU-contaminated substrates.

Growth, physiological and molecular traits in Salicaceae trees investigated for phytoremediation of heavy metals and organics.

Worldwide, there are many large areas moderately contaminated with heavy metals and/or organics that have not been remediated due to the high cost and technical drawbacks of currently available technologies. Methods with a good potential for coping with these limitations are emerging from phytoremediation techniques, using, for example, specific amendments and/or plants selected from various candidates proven in several investigations to be reasonably efficient in extracting heavy metals from soil or water, or in co-metabolizing organics with bacteria flourishing or inoculated in their rhizospheres. Populus and Salix spp., two genera belonging to the Salicaceae family, include genotypes that can be considered among the candidates for this phytoremediation approach. This review shows the recent improvements in analytical tools based on the identification of useful genetic diversity associated with classical growth, physiological and biochemical traits, and the importance of plant genotype selection for enhancing phytoremediation efficiency. Particularly interesting are studies on the application of the phytoremediation of heavy metals and of chlorinated organics, in which microorganisms selected for their degradation capabilities were bioaugmented in the rhizosphere of Salicaceae planted at a high density for biomass and bioenergy production.

Intraspecific variation of physiological and molecular response to cadmium stress in Populus nigra L.

Little is known about the variability of response to heavy metal stress within tree species, although it could be a key for a better understanding of tolerance mechanisms and for breeding. The aim of the present study was to characterize the natural variation of response to cadmium (Cd) in Populus nigra L. in order to understand the mechanisms of Cd tolerance. For that, two P. nigra genotypes, originating from contrasting environments in northern (genotype 58-861) and southern (genotype Poli) Italy, were exposed to Cd stress in hydroponics for 3 weeks. The effect of stress was estimated by measuring biomass production, photosynthetic performance and accumulation and translocation of Cd at the end of the experiment. To better understand the mechanisms of Cd tolerance, the expression of some candidate genes involved in the ascorbate-glutathione cycle (ascorbate peroxidase, glutathione reductase, glutathione S-transferase) and in metal sequestration (metallothioneins) was analyzed in leaves. Biomass production and photosynthesis were affected by the treatment in both clones but the southern clone was markedly more tolerant to Cd stress than the other. Nevertheless, the Cd content in leaves was not significantly different between the two clones and was quite low compared to other species. The content of thiols and phytochelatins (PCs), associated with the transcription profile of the glutathione S-transferase gene, indicated relevant differences in the use of the PCs pathway under Cd stress, which could explain the different tolerance to Cd. The northern clone accumulated thiols but down-regulated the GST gene, whereas the southern clone accumulated PCs and up-regulated the GST gene, which can be useful to complex and detoxify Cd. These results suggest that the glutathione pathway is involved in the differential Cd tolerance of the two genotypes. The natural germplasm of P. nigra represents a valuable resource for understanding tolerance to Cd and for selection of plant material for phytoremediation.

Screening of poplar clones for cadmium phytoremediation using photosynthesis, biomass and cadmium content analyses.

Variability of cadmium tolerance and distribution in plant organs was investigated in ten selected poplar clones from different species, hybrids and genotypes. To this end, plants were exposed for three weeks to 50 microM CdSO4 in hydroponics. Cadmium accumulation in roots, stem and leaves as well as biomass production and photosynthesis maintenance varied greatly among clones. The metal mostly accumulated in roots, up to 80% in some clones, and interestingly, only 50% in some others. In general, metal tolerance reflected the degree of photosynthesis inhibition, which was higher in clones with high cadmium accumulation in leaves. Biomass production, net photosynthesis, transpiration, and pigment content were differently reduced in cadmium-treated clones with respect to control. The variability of cadmium distribution among leaves, stem, and roots provides candidate poplars for specific phytoremediation processes of contaminated waters and soils.

Stem CO2 release under illumination: corticular photosynthesis, photorespiration or inhibition of mitochondrial respiration?

In illuminated stems and branches, CO2 release is often reduced. Many light-triggered processes are thought to contribute to this reduction, namely photorespiration, corticular photosynthesis or even an inhibition of mitochondrial respiration. In this study, we investigated these processes with the objective to discriminate their influence to the overall reduction of branch CO2 release in the light. CO2 gas-exchange measurements of young birch (Betula pendula Roth.) branches (< 1.5 cm) performed under photorespiratory (20% O2) and non-photorespiratory (< 2%) conditions revealed that photorespiration does not play a pre-dominant role in carbon exchange. This suppression of photorespiration was attributed to the high CO2 concentrations (C(i)) within the bark tissues (1544 +/- 227 and 618 +/- 43 micromol CO2 mol(-1) in the dark and in the light, respectively). Changes in xylem CO2 were not likely to explain the observed decrease in stem CO2 release as gas-exchange measurements before and after cutting of the branches did not effect CO2 efflux to the atmosphere. Combined fluorescence and gas-exchange measurements provided evidence that the light-dependent reduction in CO2 release can pre-dominantly be attributed to corticular refixation, whereas an inhibition of mitochondrial respiration in the light is unlikely to occur. Corticular photosynthesis was able to refix up to 97% of the CO2 produced by branch respiration, although it rarely led to a positive net photosynthetic rate.

Characteristics of CO2 exchange between peach stems and the atmosphere.

Gas exchange by stems is dominated by respiratory CO2 emission, but photosynthetic CO2 uptake might also occur in stem bark. We show that light-dependent CO2 uptake was present and often exceeded CO2 release by respiration in illuminated current-year peach (Prunus persica L.) stems. Respiration of peach stems, as detected by 12CO2 release into air in which the natural concentration of 12CO2 was replaced with 13CO2, was lower in the light than in the dark, but this accounted for only a fraction of the observed total CO2 uptake by illuminated stems. Stem photosynthesis was saturated at low light and was negatively affected by elevated assay temperatures (30°C), especially when combined with light intensities above saturation. An inefficient mechanism of heat dissipation by transpiration in stomata-free stems might help explain this effect. Photosynthesis was rapidly stimulated and the electron transport rate was reduced when photorespiration was suppressed by exposure to low (2 kPa) oxygen. The time-course of these changes was closely associated with a transient burst of CO2 uptake concurrent with a reduced inhibition of fluorescence yield. Photosynthesis was also stimulated by exposure to elevated (twice ambient) CO2 concentration. These combined measurements of gas exchange and fluorescence suggested that (a) photorespiration may also be active in the bark of peach stems, (b) O2 and CO2 concentrations in the bark of peach stems may be similar to ambient concentrations, (c) a large amount of electron transport unrelated to photosynthesis and photorespiration may also be present in peach stems, and (d) stem photosynthesis may be enhanced under future atmospheric conditions.

Interaction of cadmium with glutathione and photosynthesis in developing leaves and chloroplasts of Phragmites australis (Cav.) Trin. ex Steudel.

We investigated how the presence of cadmium (Cd) at the emergence of Phragmites australis Trin. (Cav.) ex Steudel plants from rhizomes interacted with leaf and chloroplast physiological and biochemical processes. About 8.5 nmol Cd mg-1 chlorophyll was found in leaves, and 0.83 nmol Cd mg-1 chlorophyll was found in chloroplasts of plants treated with 50 microm Cd. As a result, a 30% loss of chlorophyll was measured concomitantly with a comparable percentage reduction in light-saturated photosynthesis. Rubisco content and activity were lowered by 10% and 60%, respectively. Antioxidant activity was stimulated by Cd treatment and was associated with an increase in the glutathione and pyridine pools, and with a larger pool of reduced glutathione. It is suggested that the glutathione pool and its predominance in the reduced state protected the activity of many key photosynthetic enzymes against the thiophilic binding of Cd. Chloroplast ultrastructure was not significantly altered with 50 microm treatment and the efficiency of photosystem II, measured as the fluorescence ratio Fv/Fm, remained high because F0 and Fm were proportionally decreased. In plants treated with 100 microm Cd, all effects were exacerbated, but Fv/Fm remained close to that of control leaves and the glutathione and pyridine nucleotides pools were lowered. The results suggest that glutathione exerted a direct important protective role on photosynthesis in the presence of Cd.