Searching for Chemical Agents Suppressing Substrate Microbiota in White-Rot Fungi Large-Scale Cultivation.

Edible fungi are a valuable resource in the search for sustainable solutions to environmental pollution. Their ability to degrade organic pollutants, extract heavy metals, and restore ecological balance has a huge potential for bioremediation. They are also sustainable food resources. Edible fungi (basidiomycetes or fungi from other divisions) represent an underutilized resource in the field of bioremediation. By maximizing their unique capabilities, it is possible to develop innovative approaches for addressing environmental contamination. The aim of the present study was to find selective chemical agents suppressing the growth of microfungi and bacteria, but not suppressing white-rot fungi, in order to perform large-scale cultivation of white-rot fungi in natural unsterile substrates and use it for different purposes. One application could be the preparation of a matrix composed of wooden sleeper (contaminated with PAHs) and soil for further hazardous waste bioremediation using white-rot fungi. In vitro microbiological methods were applied, such as, firstly, compatibility tests between bacteria and white-rot fungi or microfungi, allowing us to evaluate the interaction between different organisms, and secondly, the addition of chemicals on the surface of a Petri dish with a test strain of microorganisms of white-rot fungi, allowing us to determine the impact of chemicals on the growth of organisms. This study shows that white-rot fungi are not compatible to grow with several rhizobacteria or bacteria isolated from soil and bioremediated waste. Therefore, the impact of several inorganic materials, such as lime (hydrated form), charcoal, dolomite powder, ash, gypsum, phosphogypsum, hydrogen peroxide, potassium permanganate, and sodium hydroxide, was evaluated on the growth of microfungi (sixteen strains), white-rot fungi (three strains), and bacteria (nine strains) in vitro. Charcoal, dolomite powder, gypsum, and phosphogypsum did not suppress the growth either of microfungi or of bacteria in the tested substrate, and even acted as promoters of their growth. The effects of the other agents tested were strain dependent. Potassium permanganate could be used for bacteria and Candida spp. growth suppression, but not for other microfungi. Lime showed promising results by suppressing the growth of microfungi and bacteria, but it also suppressed the growth of white-rot fungi. Hydrogen peroxide showed strong suppression of microfungi, and even had a bactericidal effect on some bacteria, but did not have an impact on white-rot fungi. The study highlights the practical utility of using hydrogen peroxide up to 3% as an effective biota-suppressing chemical agent prior to inoculating white-rot fungi in the large-scale bioremediation of polluted substrates, or in the large-scale cultivation for mushroom production as a foodstuff.

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.

Bismuth interaction with plants: Uptake and transport, toxic effects, tolerance mechanisms - A review.

Bismuth (Bi) is a minor metal whose abundance on Earth is estimated at 0.025 ppm. Known since ancient times for its medical properties, its use in many industrial applications has increased significantly in recent years due to its physical and chemical properties. Considered less toxic than other metals, Bi has been defined as a "green metal" and has been suggested as a replacement for lead in many industrial processes. Although the occurrence of Bi in the environment is predicted to increase, there is still a lack of information on its interaction with biota. Even though it is absorbed by many organisms, Bi has not been directly implicated in the regulation of fundamental metabolic processes. This review summarises the fragmentary knowledge on the interaction between Bi and plants. Toxic effects at the growth, physiological and biochemical levels have been described in Bi-treated plants, with varying degrees and consequences for plant vitality, mostly depending on the chemical formulation of Bi, the concentration of Bi, the growth medium, the time of exposure, and the experimental conditions (laboratory or outdoor conditions). Bismuth has been shown to be readily absorbed and translocated in plants, interfering with plant growth and development, photosynthetic processes, nutrient uptake and accumulation, and metal (especially iron) homeostasis. Like other metals, Bi can induce an oxidative stress state in plant cells, and genotoxic effects have been reported in Bi-treated plants. Tolerance responses to the excess presence of Bi have been poorly described and are mostly referred to as the activation of antioxidant defences involving enzymatic and non-enzymatic molecules. The goal of this review is to offer an overview of the present knowledge on the interaction of Bi and plants, highlighting the gaps to be filled to better understand the role of Bi in affecting key physiological processes in plants. This will help to assess the potential harm of this metal in the environment, where its occurrence is predicted to increase due to the growing demand for medicinal and industrial applications.

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.

Mathematical Model Coupled to Neural Networks Calculates the Extraction Recovery of Polycyclic Aromatic Hydrocarbons in Problematic Matrix.

Unknown extraction recovery from solid matrix samples leads to meaningless chemical analysis results. It cannot always be determined, and it depends on the complexity of the matrix and properties of the extracted substances. This paper combines a mathematical model with the machine learning method-neural networks that predict liquid extraction recovery from solid matrices. The prediction of the three-stage extraction recovery of polycyclic aromatic hydrocarbons from a wooden railway sleeper matrix is demonstrated. Calculation of the extraction recovery requires the extract's volume to be measured and the polycyclic aromatic hydrocarbons' concentration to be determined for each stage. These data are used to calculate the input values for a neural network model. Lowest mean-squared error (0.014) and smallest retraining relative standard deviation (20.7%) were achieved with the neural network setup 6:5:5:4:1 (six inputs, three hidden layers with five, five, and four neurons in a layer, and one output). To train such a neural network, it took less than 8000 steps-less than a second--using an average-performance laptop. The relative standard deviation of the extraction recovery predictions ranged between 1.13 and 5.15%. The three-stage recovery of the extracted dry sample showed 104% of three different polycyclic aromatic hydrocarbons. The extracted wet sample recovery was 71, 98, and 55% for phenanthrene, anthracene, and pyrene, respectively. This method is applicable in the environmental, food processing, pharmaceutical, biochemical, biotechnology, and space research areas where extraction should be performed autonomously without human interference.

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.

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.

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.

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.

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.

Effect of environmental stress factors on ecophysiological traits and susceptibility to pathogens of five Populus clones throughout the growing season.

The variability of ecophysiological traits associated with productivity (e.g., water relations, leaf structure, photosynthesis and nitrogen (N) content) and susceptibility to fungal and insect infection were investigated in five poplar clones (Populus deltoides Batr.-Lux clone; Populus nigra L.-58-861 clone and Populus × canadensis Mönch.-Luisa Avanzo, I-214 and Adige clones) during their growing season. The objective of the study was to determine their physiological responses under summer constraints (characteristic of the Mediterranean climate) and to propose clone candidates for environmental restoration activities such as phytoremediation. Relative water content, the radiometric water index and (13)C isotope discrimination (Δ(13)C) results reflected improved water relations in Adige and Lux during summer drought. Leaf structural parameters such as leaf area, leaf mass per area, density (D) and thickness (T) indicated poorer structural adaptations to summer drought in clone 58-861. Nitrogen content and Δ(13)C results pointed to a stomatal component as the main limitant of photosynthesis in all clones. Adige and Lux showed enhanced photoprotection as indicated by the size and the de-epoxidation index of the xanthophyll-cycle pool, and also improved antioxidant defence displayed by higher ascorbate, reduced glutathione, total phenolics and α-tocopherol levels. Photoprotective and antioxidative responses allowed all clones to maintain a high maximum quantum yield of PSII (Fv/Fm) with the exception of Luisa Avanzo and 58-861 which experienced slight photoinhibition in late spring. The study of susceptibility to rust (Melampsora sp.) and lace bug (Monosteira unicostata Muls. and Rey) infections showed Adige and Lux to be the most tolerant. Overall, these two clones presented high adaptability to summer conditions and improved resistance to abiotic and biotic stress, thereby making them highly commendable clones for use in environmental remediation programmes.

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.

Response of transglutaminase activity and bound putrescine to changes in light intensity under natural or controlled conditions in Quercus ilex leaves.

In order to further study a previously observed relationship between polyamine (PA) content and changes in irradiation, we examined the level of free and bound PAs, the activity of transglutaminase (TGase, EC 2.3.2.13) and chlorophyll fluorescence in holm oak (Quercus ilex L.) leaves in response to different levels of light intensity and amount. A diurnal trend of free and bound putrescine (F-Put and B-Put, respectively) and TGase activity was observed in plants under natural conditions in the forest, with the highest value corresponding to the maximum light intensity and amount of light received by the leaves. In another set of experiments, potted Q. ilex plants in experimental fields were subjected to a range of periods of natural photosynthetic photon flux density (PPFD) by covering or not covering the whole trees. Under a natural photoperiod (uncovered leaves), B-Put content and TGase activity paralleled the diurnal PPFD pattern, reaching a maximum at the highest PPFD; prior to this maximum, free PAs showed a significant rise. Plants that were in darkness until midday and suddenly exposed to high light intensity showed enhanced TGase activity, resulting in the maximum accumulation of B-Put. The involvement of the accumulation of B-Put reflected in the changes of the B-Put/bound spermidine ratio during the photoprotective responses to high light stress in forest plants is discussed in relation to the chlorophyll fluorescence parameters observed.