There are many methods for cleaning up contaminated soil and groundwater. Phytoremediation is an engineered method that utilizes plants and trees to remove or immobilize inorganic and organic contaminants. The plants and trees can contain contaminant plumes, uptake the contaminants, or aid in the degradation of the contaminants through several poorly understood mechanisms. Hybrid poplar trees were planted to contain a creosote contaminant plume at the study-site in Oneida, Tennessee. This research looks at how the trees will affect groundwater flow in the site. This is accomplished with the groundwater modeling program MODFLOW. The trees are simulated using the Evapotranspiration Package within MODFLOW, within the GMS modeling platform, to produce a two-dimensional unconfined aquifer viewpoint groundwater model. Site characterization, setup, and rationale are provided. The modeling methodology including calibration, sensitivity analysis, non-unique solution check, and verification are also provided. The modeling methodology included steady-state model calibration at the study-site to match observed field data; precursory steady-state and subsequent practice transient calibrations at the site; and incorporation of the simulation of evapotranspiration in the final transient model calibrations at the site. The results show that a phytoremediation system consisting of densely-planted hybrid poplar trees can indeed impact groundwater flow, although not to the extent that clearly would contain a creosote contaminant plume. Various input parameters including specific yield, transient recharge, starting heads, evapotranspiration rates, and evapotranspiration extinction depths impacted MODFLOW model sensitivity in transient calibrations. Varying the time steps in post-precipitation stress periods did not significantly impact the model output. The interception trench conductance played a minimal role in the calibration, but trench groundwater collection data was lacking, and the trench was frequently in need of maintenance. Further suggested data requirements include more frequently collected rainfall and piezometer data, as well as the installation of more piezometers outside the model domain contained in this study. Using the Evapotranspiration Package in MODFLOW provided more realistic and authentic results than using the Well Package (used in a previous study of the site by Panhorst in 2000) to simulate evapotranspiration. The Evapotranspiration Package in MODFLOW incorporates transpiration extinction depths that prevent transpiration when the water table drops below a certain depth. Further suggested program development includes incorporating an asymptotic function for transpiration rates and allowing the Evapotranspiration Package to import evapotranspiration rates, extinction depths, and elevations. It may be deduced from this impact of flow that the tree system will aid in containment of a contaminant plume, but at the trees current growth stage, and with the coal layer present at the site, the containment is limited. / Master of Science

Les travaux présentés ici portent sur les P1B-ATPases HMA2, HMA3 et HMA4 d'Arabidopsis thaliana, transporteurs de cations présents sur différentes membranes chez les plantes. L'étude du contenu cationique de plantes mutantes hma2 et hma4 a précisé le rôle important de HMA4 dans la translocation du Zn et du Cd vers les parties aériennes et sa forte affinité pour le Cd. La mesure du contenu cationique de graines de différents génotypes a montré un effet complexe des modulations de l'expression des gènes correspondants, la surexpression du gène HMA4 conduisant à une teneur en Zn de la graine similaire à celle du mutant perte de fonction. Ces résultats confirment l'importance de HMA4, et montrent la nécessité d'adapter la construction aux objectifs biotechnologiques visés. Afin de préciser le rôle de résidus conservés au sein de la famille des P1B-ATPases, j'ai étudié l'effet de l'expression d'un variant de HMA4 pour le domaine fortement conservé CPC. Les résultats obtenus in planta suggèrent une interaction avec la version native du transporteur entraînant une perte de l'activité de HMA4. Pour mener une approche structure/fonction sur ces transporteurs, L. lactis a été défini comme le meilleur candidat pour produire HMA3. Suite à l'expression de HMA3, un gain de tolérance au Cd a été observé et a permis de valider 3 variants de HMA3, mutés au niveau du pore ou du site d'hydrolyse de l'ATP, comme affectés dans l'activité de la protéine. Les membranes de L. lactis enrichies en transporteur HMA3 ou de ses variants ont permis une reconstitution in vitro en protéoliposomes permettant de mesurer une activité de transport du Cd compétitive avec le Zn et inhibée par le vanadate. / Work presented here is about Arabidopsis thaliana P1B-ATPases HMA2, HMA3 et HMA4, cation transporters found in different plant membranes. Cation content study of mutant plants hma2 and hma4 precised important role of HMA4 in upward translocation of Zn and Cd, and its high affinity for Cd. Cation content measure of seeds from different genotypes showed a complex effect of modulations of related genes expression levels, HMA4 overexpression leading to a seed Zn content similar the loss-of-function mutant one. These results confirm the importance of HMA4 and show the needs to adapt construction to biotechnological aims. To precise the role of residus conserved among the P1B-ATPases family, I studied the effect of the expression of a HMA4 variant for the highly conserved domain CPC. Obtained in planta results suggest an interaction with the native transporter leading to a loss of HMA4 activité. To perform a structure/function study on these transporters, L.lactis has been shown as the best candidate to produce HMA3. Due to HMA3 expression, a gain of Cd tolerance has been observed and allowed to validate three HMA3 variants, mutated in the pore or the ATP hydrolysis site, as affected in the protein activity. L.lactis membranes enriched with HMA3 or variants allowed an in vitro reconstitution in proteoliposomes and the measurement of a Cd transport activity competing with Zn and inhibited by vanadate.

The global soil resources are significantly threatened by pollution. In addition to the existing burden of contaminants in agricultural soils, the increasing anthropogenic input of metal(loid)s, further referred to as trace elements (TE), presents a major public health concern, since it endangers the food security of a rising human population. However, the growing demand for agricultural commodities will increase the pressure on fertile soils. In this context, steering the needed agricultural extensification towards arable TE-contaminated soils (TECS) could protect highly biodiverse or carbon stock land and, thereby, help reach global sustainability targets. The sustainable crop production on TECS requires effective and non-destructive measures to control relevant pollutant linkages. These are offered by gentle remediation options (GRO), the practical adoption of which is scarce across Europe as yet. This study provides different approaches of GRO applied to an agricultural soil in a characteristic post-mining region (Freiberg, Saxony (Germany)) in practical adoption (chapter 2) and under semi-controlled conditions (chapters 3 and 4). Due to severe topsoil contamination by metals (Cd, Pb, and Zn) and As, the pollutant linkages of concern at the study site are food-chain transfer, leaching to the groundwater, and tilling-related dust emissions. The overall aim was to find best management practices for coupling soil remediation with the production of marketable biomass. This was attempted via (i) in situ stabilization, alone (chapter 4.1) or combined with phytoexclusion (chapter 2), (ii) labile TE phytoextraction (chapter 3), and (iii) (aided) phytostabilization (chapter 4.2). Soil remediation by GRO was assessed with scientifically established (DGT, soil solution; chapter 4.1) and/or legally relevant chemical soil extractions (NH4NO3-solution; chapters 2 to 4) and pH measurements. Additionally, earthworms served as ecotoxicological endpoints (chapter 4.1). Initial soils, earthworms, vegetative and generative biomass produced from each approach, as well as the investigated soil additives, including fertilizers, were microwave-assisted chemically digested (HNO3, H2O2, aqua regia) prior to analysis. All environmental samples were analyzed for TE by inductively coupled plasma mass spectrometry (ICP-MS). Nutrient concentrations in soil additives and selected plant samples were analyzed by inductively coupled plasma optical emission spectrometry (ICP-OES). The measured TE concentrations were evaluated against applicable European and/or national thresholds. At field (chapter 2), the repeated fertilization with superphosphate and/or lime marl basically attenuated the chemical TE availability over a three years crop rotation of Brassica napus, Triticum aestivum, and Hordeum vulgare. In turn, the simultaneous phytoexclusion by low-accumulating cultivars (LAC) effectively decreased the Cd concentrations in cereal grains (by averagely 21 % in wheat and 39 % in barley). However, straw metals´ accumulation or grain As uptake partly revealed opposing trends among LAC and high-accumulating cultivars (HAC). As investigated under semi-controlled conditions (chapter 3), a sunflower (Helianthus annuus L. mutant inbred line M7 (R3B-F-U/R13M10A; test series R13F-MP)), modified towards enhanced labile TE phytoextraction by chemical mutagenesis, proved less advantageous as pre-crop for winter wheat (Triticum aestivum L. cv. Tiger) than the regionally common winter oilseed rape (Brassica napus L. cv. Lorenz). This resulted from soil alkalinization by rape, whereas sunflower mobilized more TE than it depleted from the rhizosphere. Within in situ stabilization approaches under semi-controlled conditions, a Fe- /Al-rich drinking water treatment residue (WTR), soil-applied at a rate of 1 % (m/m) prior to cultivation of wheat (Triticum aestivum L. cv. Tiger; chapter 4.1) or Szarvasi-1 (Elymus elongatus subsp. ponticus cv. Szarvasi-1; chapter 4.2), decreased the chemical availability of As, Cd, and Pb by up to 77 % , 46 %, and 61 %, respectively. Thereby, it immobilized these hazardous TE increasingly effective over time and better than a Mn-rich WTR or lime marl. The bioassays with wheat and earthworms (Dendrobaena veneta L.) showed, however, that the habitat function for biocenoses benefited more from the Fe-/Al-rich WTR when it was applied at a lower application rate (0.5 % m/m). This resulted from dose-dependent P fixation and TE entries induced by the WTR, to which Szarvasi-1 appeared insensitive. Unlike As, the availability of Cd and Zn to biota in amended soils could not be predicted by any of the applied chemical methods due to endpoint-specific binding of competing cations to the biotic ligand (plant roots, earthworm tissue), and a preferential translocation of Zn over Cd in planta. Among all studied plants, the perennials Szarvasi-1 and cup plant (Silphium perfoliatum L.; chapter 4.2) best excluded the present mixture of TE in shoots, whereby the latter exhibited growth depression. However, only grain biomass of barley and rape, and partly of low-accumulating wheat, produced at the study site presented legally compliant animal feed based on European limit values for Cd and As. Given the investigated measures´ restricted efficacy to assure forage safety, a land-use change e.g. towards the perennial Szarvasi-1, which provides a continuous plant cover at low tillage and input requirements, possibly accompanied by the monitored reuse of an Fe- /Al-rich WTR, could most promisingly control all above-stated pollutant linkages. Revenues could be generated from energy conversion or valorization in the fibrous material sector. The waste recycling of WTR in TECS, though promising, requires proper characterization, eventual process optimization, and further studies regarding long-term stability to ensure legal compliance and environmental safety. Future research and breeding efforts regarding low Cd cultivars in particular could greatly contribute to safe food or forage production at the majority of moderately contaminated sites.:Table of contents Danksagung VI Abstract VII Zusammenfassung X List of Figures XIV List of Tables XVIII Abbreviations XXII 1 Introduction 1 1.1 The contamination of agricultural soils with trace elements - a global challenge to be tackled at local scale 1 1.2 The study site 5 1.3 Objectives 7 1.4 Structure of the thesis 8 2 Field assessment of conventional fertilizers and cultivars of annual plants 11 2.1 Management of trace element-contaminated agricultural land by in situ stabilization combined with phytoexclusion over a three years crop rotation 11 3 Exploring pre-crop effects of annual oilseeds 37 3.1 Trace elements bioavailability to winter wheat (Triticum aestivum L.) grown subsequent to high biomass plants in a greenhouse study 37 4 The potentials of drinking water treatment residues and/or perennialplants 53 4.1 Trace elements bioavailability to Triticum aestivum and Dendrobaena veneta in a multielement-contaminated agricultural soil amended with drinking water treatment residues 53 4.2 (Aided) phytostabilization of trace elements using cup plant (Silphium perfoliatum L.), or tall wheatgrass (Elymus elongatus subsp. ponticus cv. Szarvasi-1) and soilapplied drinking water treatment residues 77 5 Synthesis 91 5.1 The potential of crop management practices to mitigate food chain transfer of trace elements 91 5.2 The in situ stabilization efficacy under differential experimental conditions 96 6 Future perspectives 101 References 104 List of publications 121 Erklärung 123

Microalgae play a key function in aquatic ecosystems. Their development and growth are strongly regulated by trace metals as essential elements. However, trace metals could cause negative effects when exceeding certain concentrations in the environment. In this study we tested the development and growth rate of two freshwater microalgae, the cyanobacterium Pseudanabeana mucicola and the green alga Pediastrum duplex, from Vietnam over the period of 14 days exposing to chromium (Cr) at the concentrations up to 1,936 μg L-1. Besides, the Cr uptake and absorption by P. mucicola were evaluated over 7 days incubated in medium containing 422 μg Cr L-1. The results showed that Cr at the concentrations up to 1,078 μg L-1 did not inhibit the development and growth rate of P. mucicola. Similarly, concentration of 224 μg Cr L-1 had no adverse effects on growth of P. duplex. The cyanobacterium P. mucicola could make a reduction up to 71% of Cr in the test medium, hence become a distinguished candidate for metal phytoremediation. To the best of our knowledge this is the first investigation on the responses and absorption of Cr by freshwater microalgae from Vietnam. / Vi tảo đóng vai trò quan trọng trong hệ sinh thái thủy vực. Sự sinh trưởng và phát triển của chúng được điều tiết mạnh mẽ bởi kim loại vi lượng như những yếu tố thiết yếu. Tuy nhiên, những kim loại vi lượng này có thể gây ra những ảnh hưởng tiêu cực khi vượt quá nồng độ nhất định trong môi trường. Trong nghiên cứu này, chúng tôi thử nghiệm sự phát triển và tốc độ phát triển của hai loài vi tảo nước ngọt: loài tảo lam Pseudanabeana mucicola và loài tảo lục Pediastrum duplex có nguồn gốc từ Việt Nam trong thời gian 14 ngày phơi nhiễm với crôm (Cr) tại nồng độ lên tới 1.936 μg L-1. Bên cạnh đó, sự hấp thu Cr của P. mucicola cũng đã được đánh giá trong thời gian 7 ngày nuôi trong môi trường chứa 422 μg Cr L-1. Kết quả cho thấy Cr tại nồng độ lên tới 1.078 μg L-1 không kìm hãm sự phát triển và tốc độ sinh trưởng của P. mucicola. Tương tự, tại nồng độ 224 μg Cr L-1 không có bất kì ảnh hưởng tiêu cực đến sự phát triển của P. duplex. Loài tảo lam P. mucicola có thể làm giảm 71% hàm lượng Cr trong môi trường thí nghiệm, vì vậy được xem là ứng viên sáng giá cho quá trình xử lý môi trường ô nhiễm kim loại bằng thực vật . Theo hiểu biết của nhóm tác giả, đây là nghiên cứu đầu tiên về đáp ứng và hấp thu Cr bởi những vi tảo nước ngọt có nguồn gốc từ Việt Nam.

The ability of Chromolaena odorata propagated by stem cuttings and grown for six weeks in the greenhouse to thrive in soil containing different concentrations of PCB congeners found in Aroclor and transformer oil, and to possibly remediate such soil was studied under greenhouse conditions. Chromolaena odorata plants were transplanted into soil containing 100, 200, and 500 ppm of Aroclor and transformer oil (T/O) in 1L pots. The experiments were watered daily at 70% moisture field capacity. Parameters such as mature leaves per plant, shoot length, leaf colour as well as the root length at harvest were measured. C. odorata growth was negatively affected by T/O in terms of shoot length and leaf numbers, but no growth inhibition was shown by Aroclor. At the end of six weeks of growth, Plants size was increased by 1.4 and 0.46%, but decreased at -1.0% in T/O, while increases of 45.9, 39.4 and 40.0% were observed in Aroclor treatments. Mean total PCB recoveries were 6.40, 11.7, and 55.8μg in plants tissues at Aroclor treated samples resulting in a percentage reduction of PCB from the soil to 2.10, 1.50, and 1.10 at 100, 200, and 500mg/kg Aroclor treatments respectively. There was no PCB recovery from plants in transformer oil treatments as a result of its inhibition to growth. Root uptake was found to be the probable means of remediation of PCB-contaminated soil by C. odorata, this was perhaps aided by microbes. This study has provided evidence on the ability of C. odorata to remediate PCB contaminated soil. However, the use of C. odorata for phytoremediation of PCB contaminated soil under field condition is therefore advised. / Environmental Sciences / M.Sc. (Environmental Science)

The overall efficiency of plants to remediate soils contaminated by metals depends on their growth ability especially on soils with low-fertility. For twelve weeks, the ability of Psoralea pinnata to grow well and remove chromium and iron from artificially contaminated soil was tested. The concentrations of chromium and iron in two soils obtained from different sources namely, University of South Africa premises (US) and commercial potting soil (PS) were 80 ppm, 130ppm, 180ppm, 230ppm, 280ppm, 330ppm, 380pp, 430ppm and 480ppm. Psoralea pinnata was transplanted into the contaminated soils and the experiments were watered daily to maintain 70% moisture at field capacity in a greenhouse. Shoot height and root length of Psoralea pinnata before and after planting were measured. Other parameters that were measured were number of leaves, wet shoot and dry weights, and wet root and dry weights. The growth of Psoralea pinnata, after 12 weeks of experimentation was noticeably affected by the concentrations of chromium and iron in the soil. The percentage increases in shoot height of Psoralea pinnata in the PS Soil (C-PS, 48cm from initial shoot height of 12.6cm) treatments were generally higher than the increases in the US Soil (C-US, 45.2cm from initial shoot height of 12.8cm) treatments. Psoralea pinnata in the (US) treatments accumulated Fe (50.02 ppm) from the soil more than Cr (32.38ppm). In the (PS) treatments, Psoralea pinnata also accumulated more Fe (60.57 ppm) than Cr (38.34 ppm). In the experiments containing both Fe and Cr, the US treatments with 40 ppm each of Cr and Fe, chromium was initially mostly accumulated by Psoralea pinnata (68%). At higher concentrations (320 ppm) of the combined metals (Cr and Fe) treatment, more Fe (55%) was accumulated in Psoralea pinnata. This study however showed that Psoralea pinnata may not be an efficient phytoextraction plant for hyperaccumulation. / Environmental Sciences / M. Sc. (Environmental Management)

L'utilisation des plantes pour décontaminer les sols pollués par les métaux est une solution respectueuse de l'environnement. Mais le développement de cette technique à grande échelle est encore limité en raison de l'indisponibilité de plantes avec les caractéristiques souhaitées (hyperaccumulation, biomasse élevée et croissance rapide). Les objectifs de ce travail étaient d'évaluer le potentiel de plusieurs cultivars de Pélargonium odorants pour l'extraction du Pb au champ, étudier la disponibilité du plomb en relation avec l'activité rhizosphérique et développer un protocole de transformation génétique. Parmi les six cultivars de Pélargonium odorants testés au champ, trois : Attar of Roses, Clorinda et Atomic Snowflake ont accumulé plus de 1000 mg kg-1 Pb, avec une forte biomasse. Pendant les expérimentations en conditions contrôlées, Attar of roses (le cultivar hyperaccumulateur) acidifie sa rhizosphère et augmente la concentration en COD significativement plus par rapport Concolor Lace (le cultivar non hyperaccumulateur), sans doute en réponse à la pollution métallique. Les concentrations en plomb dans les deux cultivars sont corrélées avec l'extraction au CaCl2. Les analyses par EXAFS et ESEM-EDS ont montré que le plomb présent dans les racines était principalement sous forme de complexes organiques alors que les sulfates de plomb prédominent dans le sol. Parallèlement à ces essais, un protocole de transformation génétique a été mis au point en vue de mieux comprendre les processus biochimiques impliqués dans l'hyperaccumulation et la fonction des gènes, Le système de régénération optimisé se base sur la pré-culture d'explants sur un milieu contenant 10 μM TDZ + 1 mg L-1 de chacun de BAP et NAA suivie par l'enlèvement de TDZ du milieu de culture. La kanamycine et l'hygromycine se sont avérés être de bons marqueurs sélectifs pour le Pélargonium. Deux souches d'Agrobacterium, C58 et EHA105 contenant des vecteurs binaires avec des gènes marqueurs hpt et nptII ont été choisis pour des expériences de transformation. Ils ont également le gène codant uidA séquence du gène rapporteur. Après l'infection avec C58, 4 et 107 plantes enracinées sur hygromycine ont été obtenues pour Attar of Roses et Atomic Snowflake, respectivement. Parmi ces plantes enracinées, les quatre plantes d'Attar et 82 d'Atomic Snowflake ont exprimé le Gus dans les feuilles, pétioles, les tiges et les racines comme prévu avec une séquence sous contrôle du promoteur constitutif CaMV 35S. De 20 plantes qui expriment le Gus, 7 plantes se sont avérées être positives après criblage par PCR. Après infection par EHA105, 23 et 133 plantes enracinées ont été obtenues après sélection sur kanamycine, mais aucune n'a démontré d'activité GUS. Seule des expériences d'empreintes par Southern blotting permettront de corréler le nombre d'insertions et niveau de l'expression dans ces différents événements de transformation. / Metal removal from contaminated soils using plants can provide an environment friendly solution. However, its successful application on a large scale is still limited due to unavailability of plants with desired set of characteristics i.e. hyperaccumulation, high biomass and rapid growth. The objective of this work was to assess the potential of scented Pelargonium cultivars for lead (Pb) extraction under field conditions, plant induced rhizosphere changes, soil factors influencing availability of Pb and to develop an efficient genetic transformation protocol for the selected cultivars. Of the six scented Pelargonium cultivars field-tested, three cultivars (Attar of Roses, Clorinda and Atomic Snowflake) accumulated more than 1000 mg Pb kg-1 DW, with high biomass reaching up to 45 tons ha-1 y-1 dry matter. During assays in controlled conditions, Attar of roses (Pb hyperaccumulator) significantly acidified its rhizosphere and increased Dissolved Organic Carbon (DOC) concentration as compared to Concolor Lace (non-accumulator), probably due to enhanced exudation in response to the metal stress. Lead concentrations in both cultivars were best correlated with CaCl2 extracted Pb. Extended X-ray Absorption Fine Structure (EXAFS) and Environmental Scanning Electron Microscopy-Energy Dispersive x-ray Spectroscopy (ESEM-EDS) demonstrated that Pb was mainly complexed to organic acids within plant tissues whereas the dominant form in soil was PbSO4. Parallel to the soil-plant Pb transfer assays, a genetic transformation protocol was optimized in view of better understanding biochemical processes involved in lead hyperaccumulation and gene function, in the future. The best regeneration scheme was based on the pre-culture of explants on 10 μM TDZ (Thidiazuron) in addition to 1 mg L-1 each of N6-benzylaminopurine (BAP) and α- naphthaleneacetic acid (NAA), followed by removal of TDZ from the culture medium. Kanamycin and hygromycin proved to be efficient selectable markers for genetic transformation. Two Agrobacterium strains, C58 and EHA105 harboring binary vectors carrying the selectable marker genes hpt and nptII were chosen for transformation experiments. They also contained the uidA gene coding sequence as reporter gene. After infecting with C58, 4 and 107 rooted plants on hygromycin-containing medium were obtained for Attar and Atomic cultivars, respectively. The four Attar plants and 82 Atomic plants expressed Gus in leaves, petioles, stems and roots as expected with a sequence driven by the 35S constitutive promoter. Polymerase Chain Reaction (PCR) screening was performed on Gus positive plants and 2 and 20 plants of Attar and Atomic were screened as PCR positive, respectively. After infection with EHA105, 23 and 133 rooted plants were obtained on kanamycin selection medium but none of these expressed Gus. Southern hybridization patterns will enable to correlate gene copy numbers to expression levels in these different events. The optimized protocols could be used for understanding molecular mechanisms of Pb accumulation and improvement in phytoextraction technique.

Les sols contaminés par les éléments traces potentiellement toxiques (PTTE) ont souvent des conséquences graves pour les écosystèmes terrestres. Plusieurs options de phytoremediaction ont été développées pour remédier les sols contaminés ; cependantl'efficacité et la capacité de ces techniques à réduire les concentrations excessives des éléments traces ou leur (phyto) disponibilité dans les sols contaminés doivent être évaluées Le présent travail s’est intéressé à étudier l'efficacité de deux options de de phytorémédiation, la phytostabilisation et la phytoextraction assistées par des amendements organiques et minéraux, à remédier les fortes concentrations de PTTE dans un sol naturel et dans un technosol contaminés. Les concentrations totales des éléments traces dissous ont été déterminées dans l'eau interstitielle du sol. L'intensité de l'exposition du sol a été évaluée par des capteurs DGT (gradient de diffusion dans les couches minces). Le phytodisponibilité des PTTE a été caractérisée par des tests de germination avec des haricots nains cultivés sur les sols contaminés pour lesquels les concentrations foliaires en éléments traces ont été déterminées. Ensuite un modèle de réseau de neurones artificiels a été appliqué pour comprendre les facteurs les plus pertinents sur la variabilité de la phytodisponibilité des PTTE. Les deux options ont étécapables de réduire les concentrations ou la phytodisponibilité des PTTE en présence des amendements. Les réseaux de neurones artificiels ont été très efficaces pour prédire les résultats manquants et pour déterminer les paramètres de contrôle de la variabilité de la phytodisponibilité des PTTE à partir des paramètres du sol. / Soils contaminated with potentially toxic trace elements (PTTE) often have serious consequences for terrestrial ecosystems. Several phytoremediaction have been developped to reclaim contaminated soils; however the efficiency and capacity of these techniques to reduce excessive concentrations of trace elements or their (phyto) availability in contaminated soils have to be assessed. The present work is focused on studying the effectiveness of two phyoremediation options such as phytostabilisation and phytoextraction assisted by organic and inorganic amendments to remediatethe high concentrations of PTTE in contaminated natural soils and technosoils. Total PTTE concentrations were determined in soil pore water (SPW) sampled by Rhizon soil moisture samplers. The soil exposure intensity was assessed by DGT (diffusive gradient in thin films) probes. The PTTE phytoavailability was characterized by growing dwarf beans on potted soils and analyzing their foliar PTTE concentrations. Then a model of artificial neural network was applied to understand the factors most relevant for the variability on the phytoavailability of trace elements. Both options were found to be able to reduce the concentrations or phytoavailability of PTTE in the presence of amendments. The artificial neural network has been very effective to predict missing results and to determine the control parameters of the variability of the PTTE phytoavailoability from the soil parameters.

The overall efficiency of plants to remediate soils contaminated by metals depends on their growth ability especially on soils with low-fertility. For twelve weeks, the ability of Psoralea pinnata to grow well and remove chromium and iron from artificially contaminated soil was tested. The concentrations of chromium and iron in two soils obtained from different sources namely, University of South Africa premises (US) and commercial potting soil (PS) were 80 ppm, 130ppm, 180ppm, 230ppm, 280ppm, 330ppm, 380pp, 430ppm and 480ppm. Psoralea pinnata was transplanted into the contaminated soils and the experiments were watered daily to maintain 70% moisture at field capacity in a greenhouse. Shoot height and root length of Psoralea pinnata before and after planting were measured. Other parameters that were measured were number of leaves, wet shoot and dry weights, and wet root and dry weights. The growth of Psoralea pinnata, after 12 weeks of experimentation was noticeably affected by the concentrations of chromium and iron in the soil. The percentage increases in shoot height of Psoralea pinnata in the PS Soil (C-PS, 48cm from initial shoot height of 12.6cm) treatments were generally higher than the increases in the US Soil (C-US, 45.2cm from initial shoot height of 12.8cm) treatments. Psoralea pinnata in the (US) treatments accumulated Fe (50.02 ppm) from the soil more than Cr (32.38ppm). In the (PS) treatments, Psoralea pinnata also accumulated more Fe (60.57 ppm) than Cr (38.34 ppm). In the experiments containing both Fe and Cr, the US treatments with 40 ppm each of Cr and Fe, chromium was initially mostly accumulated by Psoralea pinnata (68%). At higher concentrations (320 ppm) of the combined metals (Cr and Fe) treatment, more Fe (55%) was accumulated in Psoralea pinnata. This study however showed that Psoralea pinnata may not be an efficient phytoextraction plant for hyperaccumulation. / Environmental Sciences / M. Sc. (Environmental Management)

The ability of Chromolaena odorata propagated by stem cuttings and grown for six weeks in the greenhouse to thrive in soil containing different concentrations of PCB congeners found in Aroclor and transformer oil, and to possibly remediate such soil was studied under greenhouse conditions. Chromolaena odorata plants were transplanted into soil containing 100, 200, and 500 ppm of Aroclor and transformer oil (T/O) in 1L pots. The experiments were watered daily at 70% moisture field capacity. Parameters such as mature leaves per plant, shoot length, leaf colour as well as the root length at harvest were measured. C. odorata growth was negatively affected by T/O in terms of shoot length and leaf numbers, but no growth inhibition was shown by Aroclor. At the end of six weeks of growth, Plants size was increased by 1.4 and 0.46%, but decreased at -1.0% in T/O, while increases of 45.9, 39.4 and 40.0% were observed in Aroclor treatments. Mean total PCB recoveries were 6.40, 11.7, and 55.8μg in plants tissues at Aroclor treated samples resulting in a percentage reduction of PCB from the soil to 2.10, 1.50, and 1.10 at 100, 200, and 500mg/kg Aroclor treatments respectively. There was no PCB recovery from plants in transformer oil treatments as a result of its inhibition to growth. Root uptake was found to be the probable means of remediation of PCB-contaminated soil by C. odorata, this was perhaps aided by microbes. This study has provided evidence on the ability of C. odorata to remediate PCB contaminated soil. However, the use of C. odorata for phytoremediation of PCB contaminated soil under field condition is therefore advised. / Environmental Sciences / M.Sc. (Environmental Science)