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Many synthetic sulphonated aromatic compounds are used as starting material to produce dyes and pigments, or are released as by-products in the effluents of the textile and dye industry. A large number of these chemicals are poorly biodegradable and cannot be eliminated by classical wastewater treatment plants. To limit the impact of these pollutants on the environment, new processes, based on the use of higher plants (constructed wetlands or hydroponic systems), are under development. Detergents and surfactants are essential for both industrial and domestic applications, the most important family being the alkylbenzene sulphonates. Originally, the alkyl side chains were branched and thus recalcitrant to biodegradation. Therefore, they have been replaced by linear alkylbenzene sulphonates. Although more acceptable, present formulations still have adverse environmental and toxic effects. In this context, phytoremediation appears to be a promising approach to remove these compounds from contaminated soils and waters.

The present opinion paper is focused on the phytoremediation of organic pollutants and is based on the lectures given by the author during the International Congress of Phytoremediation of Polluted Soils, held at Vigo, Spain, 29-30 July 2014. The aim of this position paper is (1) to highlight some progress made within the last few years (after the end of COST Action 859 in October 2009) in the phytoremediation of selected organic pollutants and (2) to suggest new research and approaches, which seem important and promising in the opinion of the author to make this environmentally friendly remediation technique more attractive and more successful. Depending on the type of soil to be treated, as well as on the xenobiotic contaminants, their concentration and ageing, different approaches can be considered and are briefly presented with some recent and successful applications, but also highlighting their limitations and needs for future developments: phytoextraction of hydrophobic xenobiotic compounds like polychlorobiphenyls (PCBs), phytodegradation of xenobiotics and its possible impacts on primary and secondary metabolism of the plant and phytostimulation of rhizospheric microorganisms by root exudates for the rhizodegradation of petroleum hydrocarbons. Finally, some promising approaches are suggested for overcoming the bottlenecks and making phytoremediation a reliable, mature and sustainable technology: how to deal with mixed pollution; the potential of endophytic bacteria; possible improvements by soil amendments and co-cropping; validation of laboratory results by field experimentation; evolution of regulations from the total concentration of a pollutant to its bioavailable fraction; and the use of biomass for added-value products, fine chemicals and biofuels in biorefineries or the production of ornamental plants.

Despite recent advances in down-stream processing, production of microalgae remains substantially limited because of economical reasons. Harvesting and dewatering are the most energy-intensive processing steps in their production and contribute 20–30% of total operational cost. Bio-flocculation of microalgae by co-cultivation with filamentous fungi relies on the development of large structures that facilitate cost effective harvesting. A yet unknown filamentous fungus was isolated as a contaminant from a microalgal culture and identified as Isaria fumosorosea. Blastospores production was optimized in minimal medium and the development of pellets, possibly lichens, was followed when co-cultured with Chlorella sorokiniana under strict autotrophic conditions. Stable pellets (1–2 mm) formed rapidly at pH 7–8, clearing the medium of free algal cells. Biomass was harvested with large inexpensive filters, generating wet slurry suitable for hydrothermal gasification. Nutrient rich brine from the aqueous phase of hydrothermal gasification supported growth of the fungus and may increase the process sustainability.

A plant useful for phytoremediation has first to grow in the presence of the target pollutant without being harmed. The plant must not only be resistant to the pollutant, but must also be able to remove it from the environment and transform it into nontoxic metabolites or end products. Differences in the ability of various plant species to accumulate and metabolize particular pollutants do exist, indicating that in choosing the most appropriate species in the development of any phytoremediation process, natural biodiversity should be better explored and exploited. Plant taxonomy and phytochemistry can help in the exploitation of biochemical specificities of plants that produce natural chemicals with structures similar to xenobiotic compounds. In the case of atrazine, however, numerous results obtained for agronomical purposes are extremely useful in choosing the most appropriate families or genera for phytoremediation.

Phytoextraction with somaclonal variants of tobacco and sunflower mutant lines (non-GMs) with enhanced metal uptake and tolerance can be a sustainable alternative to conventional destructive decontamination methods, especially for stripping bioavailable zinc excess in topsoil. The overall results of a 5-year time series experiment at field scale in north-eastern Switzerland confirm that the labile Zn pool in soil can be lowered by 45-70%, whereas subplots without phytoextraction treatment maintained labile Zn concentrations. In 2011, the phytoextraction experiment site was enlarged by a factor of 3, and the labile 0.1M NaNO3 extractable Zn concentration in the soil was reduced up to 58% one period after harvest. A Mass Balance Analysis confirmed soil Zn decontamination in line with plant Zn uptake. The plants partially take Zn from the non-labile pool of the total. The sustainability of Zn phytoextraction in subplots that no longer exceed the Swiss trigger value is now assessed over time. In contrary to the phytoextraction of total soil Zn which needs a long cleaning up time, the bioavailable Zn stripping is feasible within a few years period.

Background, aim and scope: Phytoremediation does exploit natural plant physiological processes and can be used to decontaminate agricultural soils, industrial sites, brownfields, sediments and water containing inorganic and organic pollutants or to improve food chain safety by phytostabilisation of toxic elements. It is a low-cost and environment friendly technology targetting removal, degradation or immobilisation of contaminants. The aim of the present review is to highlight some recent advances in phytoremediation in the Alpine context. Main features: Case studies are presented where phytoremediation has been or can be successfully applied in Alpine areas to: (1) clean-up industrial wastewater containing sulphonated aromatic xenobiotics released by dye and textile industries; (2) remediate agricultural soils polluted by petroleum hydrocarbons; (3) improve food chain safety in soils contaminated with toxic trace elements (As, Co, Cr and Pb); and (4) treat soils impacted by modern agricultural activities with a special emphasis on phosphate fertilisation. Conclusions, recommendations and perspectives: Worlwide, including in Alpine areas, the controlled use of appropriate plants is destined to play a major role for remediation and restoration of polluted and degraded ecosystems, monitoring and assessment of environmental quality, prevention of landscape degradation and immobilisation of trace elements. Phytotechnologies do already offer promising approaches towards environmental remediation, human health, food safety and sustainable development for the 21st century in Alpine areas and elsewhere all over the world

The objective of the present study was to characterize the metabolism of Clostridium thermolacticum, a thermophilic anaerobic bacterium, growing continuously on lactose (10 g l(-1)) and to determine the enzymes involved in the pathways leading to the formation of the fermentation products. Biomass and metabolites concentration were measured at steady-state for different dilution rates, from 0.013 to 0.19 h(-1). Acetate, ethanol, hydrogen and carbon dioxide were produced at all dilution rates, whereas lactate was detected only for dilution rates below 0.06 h(-1). The presence of several key enzymes involved in lactose metabolism, including beta-galactosidase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate:ferredoxin oxidoreductase, acetate kinase, ethanol dehydrogenase and lactate dehydrogenase, was demonstrated. Finally, the intracellular level of NADH, NAD+, ATP and ADP was also measured for different dilution rates. The production of ethanol and lactate appeared to be linked with the re-oxidation of NADH produced during glycolysis, whereas hydrogen produced should come from reduced ferredoxin generated during pyruvate decarboxylation. To produce more hydrogen or more acetate from lactose, it thus appears that an efficient H2 removal system should be used, based on a physical (membrane) or a biological approach, respectively, by cultivating C. thermolacticum with efficient H2 scavenging and acetate producing microorganisms.

In the production of acetate by Clostridium thermolacticum growing on lactose, considerable amounts of hydrogen were generated. Lactose available in large amounts from milk permeate, a wastestream of the dairy industry, appears to be a valuable substrate for cheap production of biohydrogen. In this study, continuous cultivation of C thermolacticum was carried out in a bioreactor, under anaerobic thermophilic conditions, on minimal medium containing 10 g l(-1) lactose. Different dilution rates and pH were tested. C thermolacticum growing on lactose produced acetate, ethanol and lactate in the liquid phase. For all conditions tested, hydrogen was the main product in the gas phase. Hydrogen specific production higher than 5 mmol H-2 (g cell)(-1) h(-1) was obtained. By operating this fermentation at high-dilution rate and alkaline pH, the hydrogen content in the gas phase was maximized. (C) 2004 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.

Phytoremediation is an emerging technology based on the use of green plants to remove, contain, inactivate or destroy harmful environmental pollutants. Recent developments in Europe and the USA show that the approach is somewhat different on both sides of the Atlantic. In Europe, phytoremediation has more basically been research driven and, based on the outcomes, applications have been envisaged. By contrast, the approach in the USA is more application and experience driven. In spite of a growing track record of commercial success, more demonstration projects are needed to prove that phytoremediation is effective in order to rigorously measure its underlying economics, and to expand its applications. More fundamental research is also required to better understand the complex interactions between pollutants, soil, plant roots and micro-organisms at the rhizosphere level, to increase the bioavailability of pollutants, to fully exploit the metabolic diversity of plants and, thus, to successfully implement this new green technology.

Although commercial success stories abound, an urgent need still exists for more research and demonstration projects.

Sulfonated aromatic compounds are released into the environment in large amounts: the main sources of these recalcitrant pollutants are anionic detergents, dyestuffs, and their byproducts, such as ...

Sunflower mutant lines with an enhanced tolerance and metal accumulation capacity obtained by mutation breeding have been proposed for Zn, Cd and Cu removal from metal-contaminated soils in previous studies. However, soils contaminated with trace elements induce various biochemical alterations in plants leading to oxidative stress. There is a lack of knowledge concerning the metal accumulation and antioxidant responses during the growth and development of sunflowers. This study, therefore, aimed to characterise metal accumulation and possible metal detoxification mechanisms in young seedlings and adult sunflowers. Beside the inbred line, two mutant lines with an improved growth and enhanced metal uptake capacity on a metal contaminated soil were investigated in more detail. Sunflowers cultivated on a metal-contaminated soil in the greenhouse showed a decrease in shoot biomass and chlorophyll concentration in two different developmental stages. Adult sunflowers showed a lower sensitivity to metal toxicity than young seedlings, whereas mutant lines were more tolerant to metal stress than the control. Mutant lines also produced a higher amount of carotenoids on a metal-contaminated soil than on the control soil, indicating a possible protective mechanism of sunflower mutants against oxidative stress caused by Cd and excess Zn. Heavy metals primarily increased the activity of antioxidant enzymes involved in the ascorbate–glutathione cycle in sunflower leaves. Activity of dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) was strongly increased in young seedlings exposed to heavy metals. The enzyme activities were even more pronounced in mutant lines. A significantly increased ascorbate peroxidase (APOX) activity in adult sunflowers exposed to heavy metals indicated an elevated use of ascorbate after a longer exposure to metal stress. An increased antioxidant level corresponded to a high Cd and Zn accumulation in young and adult sunflowers. Metal distribution, zinc translocation in particular, from the root into the shoot tissue obviously increased during sunflower growth and ripening. Altogether, these results suggest that sunflower plants, primarily the mutant lines, possess an efficient defence mechanism against oxidative stress caused by metal toxicity. A good tolerance of sunflowers toward heavy metals coupled with an increased metal accumulation capacity might contribute to an efficient removal of heavy metals from a polluted area.

Reference LBE-ARTICLE-2002-007View record in Web of Science Record created on 2005-03-02, modified on 2016-08-08

Background, aim and scope Sulphonated anthraquinones are precursors of many synthetic dyes and pigments, recalcitrant to biodegradation and thus not eliminated by classical wastewater treatments. In the development of a phytotreatment to remove sulphonated aromatic compounds from dye and textile industrial effluents, it has been shown that rhubarb (Rheum rabarbarum) and common sorrel (Rumex acetosa) are the most efficient plants. Both species, producing natural anthraquinones, not only accumulate, but also transform these xenobiotic chemicals. Even if the precise biochemical mechanisms involved in the detoxification of sulphonated anthraquinones are not yet understood, they probably have cross talks with secondary metabolism, redox processes and plant energy metabolism. The aim of the present study was to investigate the possible roles of cytochrome P450 monooxygenases and peroxidases in the detoxification of several sulphonated anthraquinones. Materials and methods Both plant species were cultivated in a greenhouse under hydroponic conditions, with or without sulphonated anthraquinones. Plants were harvested at different times and either microsomal or cytosolic fractions were prepared. The monooxygenase activity of cytochromes P450 toward several sulphonated anthraquinones was tested using a new method based on the fluorimetric detection of oxygen consumed during cytochromes P450-catalysed reactions. The activity of cytosolic peroxidases was measured by spectrophotometry, using guaiacol as a substrate. Results A significant activity of cytochromes P450 was detected in rhubarb leaves, while no (rhizome) or low (petioles and roots) activity was found in other parts of the plants. An induction of this enzyme was observed at the beginning of the exposition to sulphonated anthraquinones. The results also indicated that cytochromes P450 were able to accept as substrate the five sulphonated anthraquinones, with a higher activity toward AQ-2,6-SS (0.706 nkat/mg protein) and AQ-2-S (0.720 nkat/mg protein). An activity of the cytochromes P450 was also found in the leaves of common sorrel (1.212 nkat/mg protein (AQ-2,6-SS)), but no induction of the activity occurred after the exposition to the pollutant. The activity of peroxidases increased when rhubarb was cultivated in the presence of the five sulphonated anthraquinones (0.857 nkat/mg protein). Peroxidase activity was also detected in the leaves of the common sorrel (0.055 nkat/mg protein), but in this plant, no significant difference was found between plants cultivated with and without sulphonated anthraquinones. Discussion Results indicated that the activity of cytochromes P450 and peroxidases increased in rhubarb in the presence of sulphonated anthraquinones and were involved in their detoxification mechanisms. Conclusions These results suggest the existence in rhubarb and common sorrel of specific mechanisms involved in the metabolism of sulphonated anthraquinones. Further investigation should be performed to find the next steps of this detoxification pathway. Recommendations and perspectives Besides these promising results for the phytotreatment of sulphonated anthraquinones, it will be of high interest to develop and test, at small scale, an experimental wastewater treatment system to determine its efficiency. On the other hand, these results reinforce the idea that natural biodiversity should be better studied to use the most appropriate species for the phytotreatment of a specific pollutant.

COST ACTION 859; International audience; PURPOSE: The term "phytotechnologies" refers to the application of science and engineering to provide solutions involving plants, including phytoremediation options using plants and associated microbes to remediate environmental compartments contaminated by trace elements (TE) and organic xenobiotics (OX). An extended knowledge of the uptake, translocation, storage, and detoxification mechanisms in plants, of the interactions with microorganisms, and of the use of "omic" technologies (functional genomics, proteomics, and metabolomics), combined with genetic analysis and plant improvement, is essential to understand the fate of contaminants in plants and food, nonfood and technical crops. The integration of physicochemical and biological understanding allows the optimization of these properties of plants, making phytotechnologies more economically and socially attractive, decreasing the level and transfer of contaminants along the food chain and augmenting the content of essential minerals in food crops. This review will disseminate experience gained between 2004 and 2009 by three working groups of COST Action 859 on the uptake, detoxification, and sequestration of pollutants by plants and consequences for food safety. Gaps between scientific approaches and lack of understanding are examined to suggest further research and to clarify the current state-of-the-art for potential end-users of such green options. CONCLUSION AND PERSPECTIVES: Phytotechnologies potentially offer efficient and environmentally friendly solutions for cleanup of contaminated soil and water, improvement of food safety, carbon sequestration, and development of renewable energy sources, all of which contribute to sustainable land use management. Information has been gained at more realistic exposure levels mainly on Cd, Zn, Ni, As, polycyclic aromatic hydrocarbons, and herbicides with less on other contaminants. A main goal is a better understanding, at the physiological, biochemical, and molecular levels, of mechanisms and their regulation related to uptake-exclusion, apoplastic barriers, xylem loading, efflux-influx of contaminants, root-to-shoot transfer, concentration and chemical speciation in xylem/phloem, storage, detoxification, and stress tolerance for plants and associated microbes exposed to contaminants (TE and OX). All remain insufficiently understood especially in the case of multiple-element and mixed-mode pollution. Research must extend from model species to plants of economic importance and include interactions between plants and microorganisms. It remains a major challenge to create, develop, and scale up phytotechnologies to market level and to successfully deploy these to ameliorate the environment and human health

Since most of the metal-hyperaccumulating wild plants only produce very low biomass and many high-yielding crops accumulate only moderate amounts of metals, the current research is mainly focused on overcoming these limitations and the optimization of metal phytoextraction. The main goal of the present study was the improvement of metal concentration and extraction properties of Helianthus annuus L by chemical mutagenesis (the non-GMO approach). Sunflowers--hybrid cultivar Salut and inbred lines-were treated with the chemical mutagen ethyl methanesulfonate (EMS). The effect of chemical mutagenesis on metal concentration in and extraction by new sunflower M1 and M2 mutants was directly assessed on a metal-contaminated field in Raft, Switzerland. Mutants of the M2 generation showed a 2-3 times higher metal shoot concentration than the control plants. The best M2 sunflower "giant mutant" 14/185/04 showed a significantly enhanced metal extraction ability: 7.5 times for Cd, 9.2 times for Zn, and 8.2 times for Pb in aboveground parts, as compared to the control plants. Theoretical calculations for the phytoextraction potential of new sunflower variants note that the best sunflower mutant can produce up to 26 t dry matter per hectare and remove 13.3 kg Zn per hectare and year at the sewage sludge contaminated site of Raft; that is a gain factor of 9 compared to Zn extraction by sunflower controls. Furthermore, the use of sunflower oil and biomass for technical purposes (lubricants, biodiesel, biogas) should produce an additional value and improve the economical balance of phytoextraction.

Sulfonated anthraquinones are precursors of many synthetic dyes and pigments, recalcitrant to biodegradation and thus not eliminated by classical wastewater treatments. In the development of a phytoremediation process to remove sulfonated aromatic compounds from industrial effluents, the most promising results have been obtained with Rheum rabarbarum (rhubarb), a plant species producing natural anthraquinones. Rhubarb is not only able to accumulate, but also to transform, these xenobiotic chemicals. Even if the biochemical mechanisms involved in the detoxification of sulfonated anthraquinones are not yet fully understood, they probably have cross talks with secondary metabolism, redox processes, and plant energy metabolism. Therefore, key problems of xenobiotic detoxification are addressed, as well as the potential role of the apoplast, often neglected, and the possible links with plant sulfur metabolism.

International audience; The resistance mechanism of vetiver (Chrysopogon zizanioides Nash) to atrazine was investigated to evaluate its potential for phytoremediation of environment contaminated with the herbicide. Plants known to metabolise atrazine rely on hydroxylation mediated by benzoxazinones, conjugation catalyzed by glutathione-S-transferases (GST) and dealkylation probably mediated by cytochromes P450. All three possibilities were explored in mature vetiver grown in hydroponics during this research project. Here we report on the role of glutathione-S-transferase in the detoxification of atrazine, as determined in vetiver leaf and root of 5-week- and 8-month-old plants grown in hydroponics. Fresh vetiver roots and leaves were cut to extract and study their GST activities toward 1-chloro-2,4-dinitrobenzene (CDNB) and atrazine, using HPLC to quantify the biosynthesis of atrazine conjugates. The global GST activity was three orders of magnitude higher than that of GST isoform able to conjugate atrazine. In vitro activities of conjugation of CDNB were similar in all root and leaf vetiver extracts, whereas activities on atrazine were only detected in leaf extracts. Entire vetiver plants exposed to C-14-atrazine were found to accumulate radioactivity at the tip of leaves under moderate transpiring conditions (75% humidity). Vetiver transformed atrazine mainly into polar compounds, identified as conjugates by TLC analyses. After 20 days of exposure, the proportion of atrazine and metabolites compared to the total penetrated radioactivity into plants was: 50% of conjugates, 28% of atrazine plus possible dealkylates and 22% of unidentified products. The maximum conjugates production was observed at the tip of leaves (29 nmol g(-1) fresh biomass), as compared to roots (6 nmol g(-1) fresh biomass). Altogether, these metabolic features indicate that conjugation to glutathione was a major metabolic pathway to detoxify atrazine in vetiver. (c) 2005 Elsevier B.V. All rights reserved.

The resistance mechanism of vetiver (Chrysopogon zizanioides) to atrazine was investigated to evaluate its potential for phytoremediation of environment contaminated with the herbicide. Plants known to metabolise atrazine rely on hydroxylation mediated by benzoxazinones, conjugation catalyzed by glutathione-S-transferases and dealkylation probably mediated by cytochromes P450. All three possibilities were explored in mature vetiver grown in hydroponics during this research project. Here we report on the chemical role of benzoxazinones in the transformation of atrazine. Fresh vetiver roots and leaves were cut to extract and study their content in benzoxazinones known to hydroxylate atrazine, such as 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)- one (DIBOA), 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) and their mono- and di-glucosylated forms. Identification of benzoxazinones was performed by thin layer chromatography (TLC) and comparison of retention factors (Rf) and UV spectra with standards: although some products exhibited the same Rf as standards, UV spectra were different. Furthermore, in vitro hydroxylation of atrazine could not be detected in the presence of vetiver extracts. Finally, vetiver organs exposed to [14C]-atrazine did not produce any significant amount of hydroxylated products, such as hydroxyatrazine (HATR), hydroxydeethylatrazine (HDEA), and hydroxy-deisopropylatrazine (HDIA). Altogether, these metabolic features suggest that hydroxylation was not a major metabolic pathway of atrazine in vetiver.

The use of green plants to remove, contain, inactivate, or degrade harmful environmental contaminants (generally termed phytoremediation) is an emerging technology. In this paper, an overview is given of existing information concerning the use of plants for the remediation of metal-contaminated soils. Both site decontamination (phytoextraction) and stabilization techniques (phytostabilization) are described. In addition to the plant itself, the use of soil amendments for mobilization (in case of phytoextraction) and immobilization (in case of phytostabilization) is discussed. Also, the economical impacts of changed land-use, eventual valorization of biomass, and cost-benefit aspects of phytoremediation are treated. In spite of the growing public and commercial interest and success, more fundamental research is needed still to better exploit the metabolic diversity of the plants themselves, but also to better understand the complex interactions between metals, soil, plant roots, and micro-organisms (bacteria and mycorrhiza) in the rhizosphere. Further, more demonstration experiments are needed to measure the underlying economics, for publicacceptance and last but not least, to convince policy makers.

Aims: The objective of this study was to increase the acetate production by Clostridium thermolacticum growing on lactose, available as a renewable resource in the milk and whey permeate from the cheese industry. Methods and Results: Experiments for increased acetate productivity by thermophilic anaerobes grown on lactose were carried out in batch cultures. Lactose at concentration of 30 mmol l(-1) (10 g l(-1)) was completely degraded by Cl. thermolacticum and growth rate was maximal. High concentrations of by-products, ethanol, lactate, hydrogen and carbon dioxide were generated. By using an efficient hydrogenotroph, Methanothermobacter thermoautotrophicus, in a defined thermophilic anaerobic consortium (58degreesC) with Cl. thermolacticum and the acetogenic Moorella thermoautotrophica, the hydrogen partial pressure was dramatically lowered. As a consequence, by-products concentrations were significantly reduced and acetate production was increased. Conclusion: Through efficient in situ hydrogen scavenging in the consortium, the metabolic pattern was modified in favour of acetate production, at the expense of reduced by-products like ethanol. Significance and Impact of the Study: The use of this thermophilic anaerobic consortium opens new opportunities for the efficient valorization of lactose, the main waste from the cheese industry, and production of calcium-magnesium acetate, an environmentally friendly road de-icer.

Sulphonated anthraquinones are known to be recalcitrant to biodegradation and are not eliminated by traditional wastewater treatment plants, leading to their accumulation in fresh water. Due to the high cost and limited efficiency of existing physical-chemical treatments, alternative cheaper processes are required to remove these compounds from industrial effluents. Four plant species were tested under hydroponic conditions for their ability to treat model effluents contaminated with mono- and disulphonated anthraquinones. Among them, Rheum rabarbarum (rhubarb) showed the most promising results and was chosen for further investigation. The apparent transpiration stream concentration factor obtained with this plant species reached up to 2.5, indicating a strong phytotreatment potential that should be further explored then exploited.

Phytoremediation is an emerging technology based on the use of green plants to remove, contain, inactivate or destroy harmful environmental pollutants. Recent developments in Europe and the USA show that the approach is somewhat different on both sides of the Atlantic. In Europe, phytoremediation has more basically been research driven and, based on the outcomes, applications have been envisaged. By contrast, the approach in the USA is more application and experience driven. In spite of a growing track record of commercial success, more demonstration projects are needed to prove that phytoremediation is effective in order to rigorously measure its underlying economics, and to expand its applications. More fundamental research is also required to better understand the complex interactions between pollutants, soil, plant roots and micro-organisms at the rhizosphere level, to increase the bioavailability of pollutants, to fully exploit the metabolic diversity of plants and, thus, to successfully implement this new green technology.

The effects in vitro of chloramphenicol on the mitochondrial electron transport of Neurospora crassa were investigated under phosphorylating, non-phosphorylating and uncoupled conditions. The antibiotic acted at more than one site of the respiratory chain: exogenous NADH dehydrogenase, Complex I, the rotenone-resistant by-pass and the alternative oxidase, although an unspecific effect could not be totally excluded.

Sulphonated anthraquinones are precursors of many synthetic dyes and pigments, recalcitrant to biodegradation, and thus contaminating many industrial effluents and rivers. In the development of a phytotreatment to remove sulphonated aromatic compounds, rhubarb (Rheum rhaponticum), a plant producing natural anthraquinones, as well as maize (Zea mays) and celery (Apium graveolens), plants not producing anthraquinones, were tested for their ability to metabolise these xenobiotics. Plants were cultivated under hydroponic conditions, with or without sulphonated anthraquinones, and were harvested at different times. Either microsomal or cytosolic fractions were prepared. The monooxygenase activity of cytochromes P450 towards several sulphonated anthraquinones was tested using a new method based on the fluorimetric detection of oxygen consumed during cytochromes P450-catalysed reactions. The activity of cytosolic peroxidases was measured by spectrophotometry, using guaiacol as a substrate. Results indicated that the activity of cytochromes P450 and peroxidases significantly increased in rhubarb plants cultivated in the presence of sulphonated anthraquinones. A higher activity of cytochromes P450 was also detected in maize and celery exposed to the pollutants. In these two plants, a peroxidase activity was also detected, but without a clear difference between the control plants and the plants exposed to the organic contaminants. This research demonstrated the existence in rhubarb, maize and celery of biochemical mechanisms involved in the metabolism and detoxification of sulphonated anthraquinones. Taken together, results confirmed that rhubarb might be the most appropriate plant for the phytotreatment of these organic pollutants.

Fermentation of milk permeate to produce acetic acid under anaerobic thermophilic conditions (approximately 60 degrees C) was studied. Although none of the known thermophilic acetogenic bacteria can ferment lactose, it has been found that one strain can use galactose and two strains can use lactate. Moorella thermoautotrophica DSM 7417 and M. thermoacetica DSM 2955 were able to convert lactate to acetate at thermophilic temperatures with a yield of approximately 0.93 g g(-1). Among the strains screened for their abilities to produce acetate and lactate from lactose, Clostridium thermolacticum DSM 2910 was found precisely to produce large amounts of lactate and acetate. However, it also produced significant amounts of ethanol, CO2 and H2. The lactate yield was affected by cell growth. During the exponential phase, acetate, ethanol, CO2 and H2 were the main products of fermentation with an equimolar acetate/ethanol ratio, whereas during the stationary phase, only lactic acid was produced with a yield of 4 mol per mol lactose, thus reaching the maximal theoretical value. When this bacterium was co-cultured with M. thermoautotrophica, lactose was first converted mainly to lactic acid, then to acetic acid, with a zero residual lactic acid concentration and an overall yield of acetate around 80%. Under such conditions, only 13% of the fermented lactose was converted to ethanol by C. thermolacticum.

In vitro breeding and somaclonal variation were used as tools to improve the potential of Indian mustard (Brassica juncea L.) to extract and accumulate toxic metals. Calli from B. juncea were cultivated on a modified MS medium supplemented with 10-200 microM Cd or Pb. Afterwards, new B. juncea somaclones were regenerated from metal-tolerant callus cells. Three different phenotypes with improved tolerance of Cd, Zn and Pb were observed under hydroponic conditions: enhanced metal accumulation in both shoots and roots; limited metal translocation from roots to shoots; reduced accumulation in shoots and roots. Seven out of thirty individual variants showed a significantly higher metal extraction than the control plants. The improvement of metal shoot accumulation of the best regenerant (3 x Cd, 1.6 x Zn, 1.8 x Pb) and metal extraction (6.2 x Cd, 3.2 x Zn, 3.8 x Pb) indicated a successful breeding and selection of B. juncea, which could be used for phytoremediation purpose.

There is a duality in plant tolerance to pollutants and its response to the pollutants’ stress. On the one hand some plants, (hyper)tolerant to heavy metals, are able to hyperaccumulate these metals in shoots, which could be beneficial for phytoremediation purposes to clean-up soil and water. On the other hand tolerant food crops, exposed to heavy metals in their growth medium, may be dangerous as carriers of toxic metals in the food chain leading to food toxicity. There is an additional duality in plant tolerance to heavy metals and that is in food crops that are tolerant and/or hyperaccumulators, which could be used on one hand for phytoremediation, under controlled conditions and on the other hand for food fortification with essential metals. Similarly, plants are also exposed to a large number of xenobiotic organic pollutants. Because they generally cannot avoid these compounds, plants take up, translocate, metabolize and detoxify many of them. There is a large variability in tolerance (defence) mechanisms against organic pollutants among plant species. This includes production of reductants but also scavenger molecules like ascorbate and glutathione and expression of the P-450 defence system, and superfamilies of the enzymes glutathione- and glucosyl-transferases. Again, with view to organic pollutants, plant detoxification mechanisms might well protect the plant itself, but produce compounds with some deleterious potential for other organisms. In this review we discuss these dualities on the basis of examples of agricultural and ‘wild’ species exposed to metal contaminants (mainly Cd) and organic pollutants. Differences in uptake and translocation of various pollutants and their consequences will be considered. We will separately outline the effects of the organic and non-organic pollutants on the internal metabolism and the detoxification mechanisms and try to indicate the differences between both types of pollutants. Finally the consequences and solutions of these dualities in plant tolerance to pollutants will be discussed.

The ability to detect early molecular responses to various chemicals is central to the understanding of biological impact of pollutants in a context of varying environmental cues. To monitor stress responses in a model plant, we used transgenic moss Physcomitrella patens expressing the beta-glucuronidase reporter (GUS) under the control of the stress-inducible promoter hsp17.3B. Following exposure to pollutants from the dye and paper industry, GUS activity was measured by monitoring a fluorescent product. Chlorophenols, heavy metals and sulphonated anthraquinones were found to specifically activate the hsp17.3B promoter (within hours) in correlation with long-term toxicity effects (within days). At mildly elevated physiological temperatures, the chemical activation of this promoter was strongly amplified, which considerably increased the sensitivity of the bioassay. Together with the activation of hsp17.3B promoter, chlorophenols induced endogenous chaperones that transiently protected a recombinant thermolabile luciferase (LUC) from severe heat denaturation. This sensitive bioassay provides an early warning molecular sensor to industrial pollutants under varying environments, in anticipation to long-term toxic effects in plants. Because of the strong cross-talk between abiotic and chemical stresses that we find, this P. patens line is more likely to serve as a direct toxicity bioassay for pollutants combined with environmental cues, than as an indicator of absolute toxicity thresholds for various pollutants. It is also a powerful tool to study the role of heat shock proteins (HSPs) in plants exposed to combined chemical and environmental stresses.

Even at low residue concentrations, persistent organic pollutants (POPs) are of increasing concern all over the world. They include numerous organochlorine pesticides, like hexachlorocyclohexane (HCH), known for its toxicity, lipophilic properties and very long turnover time. To test and settle the appropriate prevention and remediation measures, there is an imperative need to delineate suspected contaminated areas and to acquire information regarding the historical and actual production, usage and storage; the probable fate and the contamination situation of these compounds in the environment. India is a major producer and, although it officially banned HCH in 1990 for agricultural purposes, it is still permitted for the health protection programme (vector-transmitted diseases). The present report aims to address the actual contamination of soils and sediments in India with technical HCH and lindane for the period 1996-2003. The assessment was conducted using different sources of data: beside specialized scientific databases, general search engines were used. Information was also sought by visiting websites and sending a request letter to authors of publications, in charge of NonGovernmental Organizations, international organizations, companies and Indian governmental and research institutions. The appraisal revealed that, on a national level, there is a lack of information regarding lindane residues in soils and sediments. The description of parameters influencing residual concentrations of these compounds in soils is frequently omitted in these reports and the few investigations conducted do not cover the whole country, dismissing the Eastern and Central States. However, such a situation is not unique to India, and the information appears to be also scarce and patchy for many developing and developed countries worldwide. In our opinion a harmonized sampling procedure should be agreed upon at a national level and a standard procedure for lindane measurement be set at an international level. This will allow for making comparisons between different countries and improve our understanding of lindane behaviour in soil and its management. To achieve this we need enough data and obviously data of good quality, but not only this. Geographic Information System would be a very valuable tool to gather all the available information and to assess residues of HCH and other POPs at a local, regional, national and international level.

Plant biomass and metal shoot accumulation are key factors for efficient phytoextraction. In a previous study, chemical mutagenesis has been used to improve the phytoextraction potential of sunflowers. The main goal of the present study was to assess the stability of sunflower mutants with improved biomass and metal accumulation properties in the 3rd and 4th generations. As compared to control plants, the best M3 mutants showed the follwing improvement of metal extraction: Cd 3-5-fold, Zn 4-5-fold, and Pb 3-5-fod. The best M4 sunflowers also shoed enhanced metal extraction: Cd 3-4- fond, Zn 5-7-fold, Pb 6-8-fold and Cr 5-7-fold. The control sunflower inbred line IBL 04, grown directly ion the field, accumulated metals in individual organs in the following decreasing order: Cd and Zn: leaves > stem > roots > flower > seeds; Cr: roots > flower > seeds > leaves > stem. The best sunflower mutants showed either higher metal accumulation in shoots or enhanced metal accumulation in roots, suggesting to improved phytoextraction or rhizofiltration efficiency, respectively.

A combined process for carotenoids extraction and efficient bioenergy recovery from the wet microalgae biomass is proposed. High added-value products could thus be extracted prior a hydrothermal gasification of the algal biomass into synthetic natural gas. The economic sustainability of biofuel production from algal biomass as well as the large energy demands of microalgae cultivation and harvesting is addressed in this paper. Two green solvents, ethanol and 2-methyltetrahydrofuran (MTHF), were used to achieve the maximum extractability of selected carotenoids. Pure MTHF was tested for the first time as an alternative renewable solvent for carotenoid extraction from wet biomass and promising results were obtained (30 min at 110 degrees C), with 45% of total carotenoids being extracted. The energy content of the residual biomass corresponds to a High Heating Value (HHV) of 18.1 MJ/kg. With a 1: 1 mixture of both MTHF and ethanol, more carotenoids were extracted from wet biomass (66%) and the remaining HHV of the residual biomass was 15.7 MJ/kg. The perspectives of combined carotenoid extraction and energy recovery for a better microalgae valorization are discussed.

Sunflower can be used for the remediation of metal-contaminated soils. Its high biomass production makes this plant species interesting for phytoextraction and using sunflower oil for a technical purpose may improve the economic balance of phytoremediation. The aim of the present field study was to screen 15 commercial cultivars of Helianthus annuus L., grown on metal-contaminated soil, to find out the variety with the highest metal extraction, which can be further improved by mutation or in vitro breeding procedures. Two different fertilizers (ammonium sulphate and ammonium nitrate) were also used to enhance the bioavailability of metals in soil. Highly significant differences were observed within tested varieties for metal accumulation and extraction efficiency. Furthermore, ammonium nitrate increased cadmium extraction, whereas ammonium sulphate enhanced zinc and lead uptake in most tested cultivars. In this field-based sunflower screening, we found enhanced cumulative Cd, Zn, and Pb extraction efficiency by a factor 4.4 for Salut cultivar. We therefore emphasize that prior to any classical breeding or genetic engineering enhancing metal uptake potential, a careful screening of various genotypes should be done to select the cultivar with the naturally highest metal uptake and to start the genetic improvement with the best available plant material.

Reference EPFL-ARTICLE-159257doi:10.1007/s11356-009-0242-1View record in Web of Science Record created on 2010-11-30, modified on 2016-08-09